Archives of Razi Institute
Volume:78 Issue: 3, May-Jun 2023

Coxiella burnetii (C. burnetii), the etiological agent of the Q fever disease, ranks among the most sporadic and persistent global public health concerns. Ruminants are the principal source of human infections and diseases present in both acute and chronic forms. This bacterium is an intracellular pathogen that can survive and reproduce under acidic (pH 4 to 5) and harsh circumstances that contain Coxiella-containing vacuoles. By undermining the autophagy defense system of the host cell, C. burnetii is able to take advantage of the autophagy pathway, which allows it to improve the movement of nutrients and the membrane, thereby extending the vacuole of the reproducing bacteria. For this method to work, it requires the participation of many bacterial effector proteins. In addition, the precise and prompt identification of the causative agent of an acute disease has the potential to delay the onset of its chronic form. Moreover, to make accurate and rapid diagnoses, it is necessary to create diagnostic devices. This review summarizes the most recent research on the epidemiology, pathogenesis, and diagnosis approaches of C. burnetii. This study also explored the complicated relationships between C. burnetii and the autophagic pathway, which are essential for intracellular reproduction and survival in host cells for the infection to be effective.

Rosemary Leaves (Rosmarinus officinalis) gained importance as natural antioxidants which strengthen the endogenous antioxidant defenses through die. The present experience was designed to assess the protective effect of ethanolic extract of rosemary leaves on the adrenal gland and testicular toxicity in male rabbits exposed to Cypermethrin. Forty healthy male rabbits were distributed into four groups of 10 animals each; the animals were administered cypermethrin 66.5 mg/kg alone or concurrent with Rosemary extract in both dosages (100 and 200 mg/kg) for 45 days, and the blood samples were taken from all animals for estimation hormones indices, the Anaesthetized animals were euthanized and adrenal gland and testes were separated for histopathological analysis. Results revealed that the exposure to Cypermethrin induced stress and infertility as evidenced by elevation in the level of cortisol concurrently with a lowering in ACTH level. Also, recording elevation in FSH and LH levels and a significant decline in estradiol level related to a reduction in testosterone levels observed noticeable compared to healthy control. While Concurrent exposure to Cypermethrin and Rosemary extract significantly improved hormone criteria compared to rabbits exposed to Cypermethrin alone. Histological lesions in this study include: the adrenal gland appeared thick fibrous capsule surrounding the adrenal tissue, destruction of adrenal cortex and vacuolation of three layers of the cortex, while in testes marked inhibition of spermatogenesis and degeneration of Sertoli cells with few numbers of Leydig cells were shown. These alterations were brought about by cypermethrin toxicity, while the treatment of Rosemary leaves extract with Cypermethrin alleviated the deleterious effect of Cypermethrin on the adrenal gland and testes and also restored spermatogenesis. The results showed that the extract of rosemary leaves possesses anti-infertility and strong antioxidant activities and can be used as a fertility-increasing drug to control sexual hormones also spermatogenesis, preventing toxicity and its pathophysiological consequences.

Because of the mutual relationship between neural inflammation and seizure, this study aimed to determine the effects of intracerebroventricular (ICV) injection of the steroidal and non-steroidal anti-inflammatory drugs on pentylenetetrazol (PTZ)-induced seizures during the estrous cycle in rats. A total of 105 adult female Wistar rats were selected and divided into seven groups, including the control (saline), ketorolac tris salt (7.5, 15, and 30 µg), and methylprednisolone acetate (0.15, 0.3, and 0.6 µg), each with four subgroups (proestrus, estrus, metestrus, and diestrus) and three replicates (n=5). After a week of acclimatization, the estrous phase determination and synchronization were performed. Acute epilepsy was inspired by the intraperitoneal injection of 80 mg/kg of PTZ 30 min after the ICV injection of ketorolac and methylprednisolone acetate. The initiation time of myoclonic seizures (ITMS), the initiation time of tonic-clonic seizures (ITTS), seizure duration (SD), and mortality rate (MR) were measured for 30 min. Data were shown as mean±SD and analyzed using One-way ANOVA followed by Tukey–Kramer multiple comparison post hoc test (P<0.05). According to the results, ketorolac (15 and 30 µg) and methylprednisolone acetate (0.3 and 0.6 µg) significantly increased the ITTS and ITMS but decreased SD during the estrous cycle, compared to the control (P<0.05). Moreover, MR and SD were significantly decreased by ketorolac (7.5, 15, and 30 µg) and methylprednisolone (0.3 and 0.6 µg), compared to the control during the estrous cycle (P<0.05). Therefore, it seems that both ketorolac and methylprednisolone possess dose-dependent anticonvulsant effects that may decrease neural inflammation.

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). The laboratory diagnosis of the disease includes various bacteriologic and immunologic methods. Despite the effectiveness of many of these methods in diagnosing active TB, their high cost and time-consuming nature have led researchers to adopt more accurate and rapid screening methods based on specific antigens for M. tuberculosis. The present study aimed to measure specific antibody serum levels against the early secretory antigenic target 6 kDa (ESAT-6) recombinant protein in healthy people and compare it to TB patients. The target population included 27 TB patients and 87 healthy individuals with no clinical TB symptoms. The healthy population was divided into two groups, including positive purified protein derivative (PPD) and negative PPD (35 and 52 people, respectively), using the Tuberculin skin test. The specific antibody level against the ESAT-6 recombinant antigen and the PPD protein was measured using an indirect Enzyme-Linked Immunosorbent Assay (ELISA) test. The results of the study showed that the majority of the healthy population with no symptoms of clinical TB and having negative skin test results did not have antibodies against the recombinant ESAT-6 (98%) and PPD (96%) antigens. On the other hand, there was a high level of the specific antibody of the ESAT-6 recombinant and PPD antigens in TB patients (77%). It is notable that in people with positive skin test results, the level of the antibody against the ESAT-6 recombinant antigen and PPD antigen was 94%. The results demonstrated that the ELISA method based on the measurement of antibodies against the ESAT-6 recombinant antigen can be a proper diagnostic method for rapid and accurate screening of healthy from infected people.

One of the most important nosocomial organisms that cause urinary tract infections (UTIs) in cancer patients is Escherichia coli. A significant cause of concern in managing UTIs is the development of carbapenem-resistant bacteria. Escherichia coli with carbapenem resistance has become a more serious problem, particularly in Iraq. In this regard, the present study aimed to estimate the prevalence of carbapenem-resistant E. coli in Al-Basrah, Iraq. Conventional tests and the Vitek®2 system were used to identify the isolates and determine the susceptibility of E.coli isolates to antimicrobials. In addition, E.coli isolates were tested by mCIM and eCIM methods. Moreover, the major carbapenemase genes, including blaSPM, blaIMP, blaVIM, and blaKPC were detected by polymerase chain reaction. In total, 120 urine samples were collected from cancer patients who were suspected of having urinary tract infections at Basrah Center of Oncology Al-Sader Teaching Hospital, Basrah, Iraq. Identification of bacterial growth by using biochemical tests revealed different bacterial species. The most frequent bacteria were E. coli (n=22, 53.65%) isolates. The results showed that 13 (59.09%) and 11 (50%) out of 22 E. coli isolates were positive for the production of carbapenemase, based on the eCIM and sCIM, respectively. The majority of E.coli in this study possessed the blaVIM gene (n=13, 59.1%), followed by the blaKPC gene (n=5, 22.73%), blaIMP gene (n=5, 22.73%), and blaSPM gene (n=4, 18.18%). There is a spread of more than one type of carbapenemase among the E. coli isolates collected from UTI cancer patients in Basrah Hospital. The E. coli identified in the current study had a strong capacity to produce carbapenemase enzymes against the four generations of antibiotics, including imipenem and meropenem antibiotics.

Probiotics have been used for over a century to prevent and treat diseases. They can reduce the effects of gastroenteritis and are now used to treat acute diarrhea. This study aimed to evaluate the co-aggregative effects of probiotics bacteria against diarrheal causative bacteria. For this purpose, 11 isolates of probiotic bacteria were used in the current study, including three Lactobacillus plantarum, one Lactobacillus gasseri, two Lactobacillus fermentum, three Lactobacillus acidophilus, and two Lactococcus garvieae isolates. All isolates were tested for antibiotic susceptibility, autoaggregation ability, adhesion ability, antibacterial activity, acid tolerance, and bile salts tolerance. The results showed that most of them had the ability to autoaggregate after 4 h, with the highest percentage of 57.14% for L. fermentum. For the antibiotic susceptibility test, all the isolates showed resistance against trimethoprim/sulfamethoxazole, except one isolate. Moreover, all the isolates, except one, were susceptible to both vancomycin and tetracycline. All tested isolates had adhesion ability with different survival rates, which reached 34.57% for L. plantarum in acidic conditions. Besides, the highest survival rate was 85.17%, which belonged to L. garvieae, for bile salt tolerance. Probiotic isolates had an antibacterial effect against diarrhea-causative bacteria with an inhibition diameter of 17-49 mm for different Lactobacillus spp. and Lactococcus spp. isolates. Furthermore, the co-aggregation ability of probiotic isolates against diarrhea-causative bacteria was studied, and results showed that probiotic isolates had a co-aggregative effect against diarrhea-causative bacteria, Escherichia coli, Shigella sonnei, and Providencia alcalifaciens, after 24 h of incubation. The highest co-aggregative effect of probiotics isolates belonged to L. fermentum and L. acidophilus against P. alcalifaciens with a co-aggregation percentage of 100%, while the lowest co-aggregation rate was 14.29% against E. coli.  The findings revealed the probiotic properties and co-aggregative effects of probiotic bacteria against diarrhea-causative bacteria.

This report aimed to determine the effect of lipopolysaccharide (LPS) on food intake in broiler chicks with different rations. All birds received a starter diet until five days of age, but experimental diets were provided on days of injections. In experimental group one, chickens received an intracerebroventricular (ICV) injection of LPS (25, 50, and 100 ng) with a standard diet. In experimental group two, chickens received intraperitoneal (IP) injections of LPS (50, 100, and 200 µg) with a standard diet. In experimental group three, birds received ICV injections of saline and different diets. Accordingly, a standard diet without fat, a diet containing 20% higher nutrient energy than the standard, a diet containing 20% less nutrient energy than the standard, and a standard diet containing fat were offered to them to investigate the desire of chickens for the diets. Experimental groups four, five, and six were similar to experimental group three, except that the chickens received ICV injections of LPS. In experimental groups seven, eight, and nine, chickens received IP injections of LPS with different diets. Afterward, their cumulative food intake was measured until 180 min post-injection. According to the results, ICV and IP injections of LPS decreased food intake (P<0.05). However, the ICV injection of saline increased the desire of chickens for the standard diet with fat (P<0.05). The ICV injection of the LPS (50 and 100 ng) increased the appetite for a standard diet with nutrient energy 20% higher than the standard and a standard diet containing fat, at 120 and 180 min after the injection (P<0.05). In addition, IP  injection of LPS (200 µg) significantly increased the desire for a standard diet with nutrient energy 20% higher than the standard and a standard diet containing fat (P<0.05). These results suggested the desire of chickens for different types of rations is affected by central or peripheral administration of the LPS.

Ischemic heart disease (IHD) is a common diagnosis and a leading cause of death in both males and females. It accounts for 30% of deaths worldwide, including 40% in high-income countries and approximately 28% in developing nations. Several cardiac markers have been used to diagnose and manage cardiovascular diseases. The Coenzyme Q10 (CoQ10) plays a potential role in the prevention and treatment of cardiovascular diseases by improving cellular bioenergetics. This study aimed to evaluate the role of CoQ10 and other biochemical parameters in IHD (angina pectoris and myocardial infarction). A case-control study was conducted at the Intensive Care Unit of Ibn-Sina Teaching Hospital and Al-Salam General Hospital in Nineveh Province, Iraq, for two months, from April 1 to June 1, 2022. It included 90 adult participants divided into case and control groups. The case group included 60 patients admitted to the Intensive Care Unit and diagnosed with IHD (myocardial infarction or angina pectoris), and the control group included 30 healthy participants matched in age and gender with the case group. Subsequent assay of C-reactive protein (CRP), creatine phosphokinase (CPK), troponin level, and serum CoQ10. In this study, 81.7% of patients in the case group were diagnosed with myocardial infarction. Means of serum lactate dehydrogenase (LDH), CRP, CPK, and troponin were significantly higher, while those of CoQ10 were significantly lower in the case group, compared to the controls. Statistically, a significant moderate negative correlation was detected between CoQ10 level and age. Moreover, significant weak correlations were observed between CoQ10 level and all serum LDH, CRP, and troponin levels. Patients with IHDs had considerably low serum levels of CoQ10, compared to the control group. The highest mean value of lipid profile, except for triglyceride, was observed in patients with IHD, compared to the control group. This explains the role that cholesterol compounds play in the progression of IHD. No significant correlations were found between CoQ10 with body mass index and CPK. The CoQ10 had a negative correlation with age, serum LDH, CRP, and troponin.

Infectious bursal disease virus (IBDV) causes a highly contagious disease associated with immunosuppression in young chickens. Production of either egg-based or primary cell-based high-quality vaccines requires time-consuming and costly procedures. To determine a suitable cell line for IBDV replication, L929 cell line was a candidate for the growth kinetics processing of the virus. The L929 cells were proliferated in monolayer, and doubling time was calculated. Replication kinetics an IBDV isolate at the multiplicity of infection 0.1 PFU/cell were determined using virus titration. To adapt IBDV on L929 cells, seven consecutive passages were performed. Virus titer and levels of apoptosis were quantitatively analyzed at each passage. The viral VP2 gene was amplified and sequenced in three passages. An average doubling time of 21 h was estimated for monolayers of L929 cells. Although during early passages, virus growth did not produce a clear cytopathic effect (CPE), an increase in IBDV titers was observed. Serial passages led to the evidence of marked CPEs and an increase in the virus titer in the third passage. During the fourth to seventh passages, consistent CPEs characterized by the formation of granulated and round cells were evident within 24 to 48 hours post-inoculation. The titer of the virus was increased in the third passage onwards to peak in the fourth and constant at 5.9 TCID50 until the end passage. The IBDV replication in connection with DNA fragmentation and FITC, revealed the characteristic picture of apoptosis in a time-dependent manner. We found that the IBDV could easily be adapted to L929 cells, increasing virus yields by about two orders of magnitude. These results indicated that the cell line may be useful in the production of efficient virus particles.

Fowl Adenoviruses (FAdVs) are widely distributed pathogens across the globe. The FAdVs from serotypes FAdV 2, 3, 8a, 8b, 9, and 11 are responsible for inclusion body hepatitis (IBH). Recently, increased mortality and IBH-suspected lesions were observed in 8-10-day-old broiler chickens in West Azerbaijan Province, Iran. In this regard, the present study aimed to compare penton and hexon genes of ADDV11 in the molecular detection of IBH in broiler chickens. In total, 100 liver specimens were collected from 10 suspected farms, and their DNAs were extracted. Two polymerase chain reactions (PCRs) were applied; one targeting the L1 region of the hexon gene and another aiming at the penton gene. Based on the findings, 60% of samples showed positive results in both PCRs and phylogenetic analysis clustered the studied viruses into serotype 11 (species D) FAdV. The detected FAdVs also shared a multitude of homologies with previously published serotype 11 viruses from Iran and those identified in Pakistan, Saudi Arabia, India, China, and Canada. This research not only provides an update on circulating FAdVs in Iran, but also introduces the penton gene as an alternative target for IBH diagnosis. Considering that IBH is a primary disease in Iran with both horizontal and vertical routes of transmission, urgent preventive measures are needed.

Validation is a Good Manufacturing Practice principle that proves any procedure, process, method, equipment, material, activity, or system actually leads to the expected results. This study validates the method for the determination of free formaldehyde in biological products (including the diphtheria-tetanus vaccine and tetanus toxoid antigen). The operating procedure of this method is based on pharmacopoeial monographs. It also does not require full validation, although its suitability under the actual condition of use should be verified. Performance characterizations, such as accuracy, intra-precision (repeatability), intermediate precision (inter-precision), linearity, range, and the limit of quantitation, were investigated and calculated. Accuracy and precision were studied at different concentration levels by spiking known amounts of formaldehyde in real samples. The accuracy and precision results were expressed as the recovery and the relative standard deviation (RSD), respectively. Precision was expressed as intra-precision (repeatability) and inter-precision. Intra-precision or repeatability was performed by one operator in one day by adding three levels of concentration to the products. The inter-precision was conducted by one operator in three individual days within the same laboratory at three concentration levels. Range and linearity were assessed by investigating the correlation coefficient of the regression line between different concentrations of formaldehyde and their response. The acceptance criteria and limits were defined for these validation parameters in these biological products. The RSD for intra-day and inter-day precision studies was less than 5% in a medium concentration of linear range. At this concentration level, accuracy was 90%-110%. The method’s linearity ranged between 0.0000039%-0.01% w/v of formaldehyde with a correlation coefficient of 0.9999. The results exhibited sufficient linearity, accuracy, precision, and range. Therefore, this method can be used successfully to determine free formaldehyde for biological products.

The present study aimed to evaluate the implantation of decellularized small intestinal submucosa- extracellular matrix )SIS-ECM( seeded with bone marrow mesenchymal stem cells (BM-MSCs) to repair full-thickness Achilles tendon defect. For this purpose, 20 healthy adult stray dogs aged 8-12 months old (15±3 kg of weight) were enrolled in this study under an aseptic environment and general anesthesia. A 1.5 cm-long segment-based resection was performed in the mid-substance of the Achilles tendon in the control group (n=10) that did not receive treatment. While, in the experimental group (n=10), regarding the defect of the tendon, the stumps were bridged with decellularized SIS seeded with BM-MSCs (5×106) cells implanted. Afterward, the stumps of the tendon were sutured using the modified Kessler technique (4-0) polypropylene thread. The biomechanical observations of the tendon defect showed an increase in the tensile strength in the experimental group, compared to the control animals. It should be mentioned that this difference was significant (P≤0.05). Histopathological observations of biopsies harvested after the 4th, 8th, and 12th weeks revealed that the implanted graft had seeded with MSCs enhanced high-quality cellular infiltration and the host tissue healing was improved. Similar to the normal tendon, a dense organized collagenous tissue with high cellularity and vascularity was observed due to the presence of the remodeled ECM. However, the arrangement of collagen-fiber-derived connective tissue appeared to be more dominant than that in the experimental group, with less adhesion in the 12th week post-treatment. These findings suggest that the BM-MSCs inoculated with SIS can be employed to repair a damaged Achilles tendon due to the fact that this combination enhances the regeneration of the affected tendon.

Four different propolis samples obtained from different regions of Iran were evaluated for their antibacterial effects against the bacterial agents responsible for two important honeybee diseases. Paenibacillus larvae (P. larvae) and Melissococcus plutonius (M. plutonius), as the etiological agents of American foulbrood (AFB) and European foulbrood (EFB) diseases, were subjected to propolis ethanolic extracts in the agar well diffusion assay. The minimum inhibitory concentrations (MIC) and the minimum bactericidal concentrations (MBC) of the antibacterial effects of the samples against the two indicator organisms were determined by the microdilution technique using different concentrations of the propolis extracts. Finally, the synergistic antibacterial actions of the mixed propolis samples were determined, and their MIC and MBC values were recorded. A two-way analysis of variance was used to evaluate correlations among the diameters of the inhibition zones, the bacterial agents, and the propolis extracts. Based on our results, three of the propolis samples showed significant antibacterial effects against P. larvae and M. plutonius during the agar well diffusion assay. Furthermore, the antibacterial capacity of the propolis samples, when mixed in equal proportions, was significantly enhanced, as indicated by the obtained MIC and MBC values. Approximately, 0.02 mg/mL of mixed propolis samples was required for inhibiting the growth of both pathogens. A direct correlation was observed between propolis concentrations and their antibacterial activity. The results of the study are conclusive of the significant antibacterial actions of Iranian propolis samples against the etiological agents of the mentioned honeybee diseases, suggesting their probable use as a safe biological agent to control AFB and EFB diseases.

Foot and mouth disease (FMD) and enterotoxemia are important diseases of hoofed animals. Vaccination against livestock pathogens, especially these two diseases, plays a key role in the prevention and control of these diseases. The use of combined vaccines with the aim of creating a better immune response and producing cheaper vaccines is a great contribution to Vaccine industry. This research aimed to compare the immunogenicity of FMD (O) and Clostridium perfringens type B toxoid along with adjuvant (MF59) and Montanide (ISA70) to create the best immunogenicity. To investigate the immune responses of vaccines, it was injected into an animal model, and the antibody titer was measured by enzyme-linked immunosorbent assay (ELISA) test and VN antibody titer. The results showed that the formulation with MF59 adjuvant brought more stable immunogenicity against FMD and Clostridium perfringens type B, and the length of the immunogenicity period also increased significantly. Therefore, the combined vaccine (Clostridium perfringens + FMD) could play a major role invaccine industry as an alternative vaccine against Clostridium perfringens and FMD in livestock.

Goats are the earliest domesticated ruminants. The local goat, Capra hircus, is considered one of the most important animals globally to provide good livestock production under harsh environmental conditions. This study aimed to detect the genetic structures of the local Iraqi goats bred in the central and southern regions of the country and investigate the possibility of benefiting from their genetic structures to construct improvement programs for increasing the productivity of these animals. To this end, blood samples were taken from 15 domestic black goats. A total of 10 ml of each animal’s blood was placed in plastic containers of 10 ml. The DNA was extracted and sent to the laboratories of Juan Ju University, People’s Republic of China, to analyze the sequences of the nitrogenous bases of the Cytochrome b (Cytb) gene. The results showed the presence of a genetic morphology for a segment of 670 base pairs for all the studied samples, and 15 sequences of this strain were recorded in the gene bank under the following accession numbers (LC496353.1:1-LC496367.1:1). The sequences of the nitrogenous bases of this segment of the gene, which were registered in the gene bank of some international goat breeds, were used for comparison with the sequences of black Iraqi goats to analyze the phylogenetic tree, calculate the genetic distance, study haplotypes, and calculate neutrality. The results showed the presence of one mutation in the studied segment of the Cytb gene, with a size of 670 bp. The mutation in base 46 of the studied gene converted from the purine group to the pyrimidine group (the shift from the nitrogen leaders A<C) in all the studied samples. It led to the transformation of the amino acid Asparagine into Histidine, where 233 amino acids were obtained, dominated by the amino acid Isoleucine and Leucine, over other amino acids at a rate of 14.34% and 11.21%, respectively. The phylogenetic tree showed the existence of two main branches, one of which included the local black Iraqi goat breed, and the other included all the international breeds under comparison. It is concluded that the black Iraqi goat breed has a different origin from other breeds.

Aldosterone is a key component of Renin-Angiotensin-Aldosterone System (RAAS). The RAAS could play a substantial role in the pathophysiology of coronavirus disease 2019 (COVID-19). Moreover, the dynamics of the Hypothalamic-Pituitary-Adrenal (HPA) axis may have changed in COVID-19. Cortisol, as an important factor in assessing immune system activity, is an important part of this axis. The present study compared the serum levels of aldosterone and cortisol in COVID-19 outpatients with those of potentially non-infected participants. It was also aimed to assess the possible association between serum levels of aldosterone and cortisol with clinical symptoms progression in COVID-19 outpatients. Demographic characteristics (i.e., gender and age) and clinical data (i.e., oxygen saturation [SPO2], respiratory rate [RR], and heart rate) were collected. Serum cortisol and aldosterone measurements were conducted using the ELISA technique. Clinical symptoms of the positive polymerase chain reaction (PCR) group were followed up on for 28 days in weekly intervals. SPO2 was significantly lower in the positive PCR group; however, the RR was significantly higher (P=0.03 and P=0.001, respectively). Significantly higher levels of aldosterone were found in males of the negative PCR group, compared to females (P=0.05). Cortisol (OR=0.937, P=0.033) and aldosterone (OR=1.005, P=0.020) levels had a decreasing and increasing effect on the chances of respiratory symptoms occurring over time, respectively. Furthermore, over time, women were twice as likely as men to develop neurologic symptoms (OR=0.530, P=0.015). According to the findings of this study, cortisol and aldosterone are associated with the chance of respiratory symptoms occurring over time. However, the levels of these two markers do not seem to be related to the progression of clinical symptoms of lower grades of COVID-19.

The SARS-CoV-2 virus, which emerged in December 2019, has infected millions worldwide and caused many deaths. Due to its high mortality rate, several studies assessed the effectiveness of different drugs against COVID-19, mainly in reducing the hospitalization rate among the elderly and compromised patients. Lopinavir-ritonavir combination and remdesivir were among the medications used to treat COVID-19. Due to considerable differences in the effectiveness and clinical outcomes of the two treatments, this study aimed to compare the clinical outcomes between COVID-19 patients treated with antiretrovirals (lopinavir-ritonavir) and remdesivir. A total of 33 patients on lopinavir-ritonavir and 35 on remdesivir were selected for this study. A retrospective comparative analysis was conducted based on demographic characteristics, hospital stay, laboratory parameters of C-reactive protein (CRP) and plasma blood oxygen saturation (SPO2), clinical treatment, and a clinical outcome assessment extracted from hospital archive data. Both treatments improved patient outcomes, yet there was a significant difference between lopinavir-ritonavir and remdesivir groups in platelet count, CRP, SPO2, and monocyte results, with remdesivir showing better clinical outcomes. No significant difference was reported in white blood cells, lymphopenia, and lactate dehydrogenase between the two treatments. It is still necessary to conduct further research to determine how effective the two treatments are in treating severe COVID-19 cases due to the limited number of available studies and the inconsistency in research methods and measurements.

Tendon is similar to rope and consists of strong, flexible, dense connective tissue. Tendon disorder healing is challenging as it is an avascular tissue. The repaired tissue appears scar-like, and its biomechanical properties never ultimately return to their pre-injury state. This study aimed to evaluate the effects of platelet-rich fibrin (PRF) on the hydroxyproline content of the Achilles tendon after injury. For this purpose, 24 adult rabbits weighing 1.5-2 kg were used in this study. The animals were divided into three groups of eight, including the advanced-PRF (A-PRF) group in which the tendon defect was treated with xenogeneic A-PRF, the leukocyte-PRF (L-PRF) group in which xenogeneic L-PRF was used for tendon defect treatment, and a control group which was treated with normal saline. Hydroxyproline concentration was measured 1 and 2 months after the operation. Clinically, lameness was improved in the A-PRF group, compared to the L-PRF and control groups at the end of the third week after the surgery. Hydroxyproline level was significantly increased in the A-PRF group (50.33±1.44), compared to the L-PRF (44.70±1.12) and control (35.97±1.05) groups 2 months after the surgery (P<0.05). Moreover, the L-PRF group showed an increase in hydroxyproline content, compared to the control at the same period. The results of the current study demonstrated that A-PRF could enhance the hydroxyproline content of rabbit Achilles tendon after injury. Xenogenic PRF can be used as an alternative biomaterial to accelerate and regenerate tendon tissue.

Duchenne muscular dystrophy (DMD) is an X-linked recessive neuromuscular illness with a progressive course caused by mutations in the gene encoding the protein dystrophin (DMD; locus Xp21. 2). This study aimed to investigate the clinical aspects of DMD progression according to the mutation type. Included in the study were 38 boys aged 3 to 11 years. Laboratory (biochemical evaluation of the level of creatinine phosphokinase, multiplex ligation-dependent probe amplification [MLPA], and next-generation sequencing [NGS] analysis of the DMD gene), genealogical, and clinical approaches were utilized (including adapted Hammersmith Functional Motor Scale, the study of auditory-speech memory by the "Memorizing 10 words" method, and a general neurological assessment). The MLPA revealed deletion in 22 cases (57.8%), duplication in 6 (15.7%), and negative results in 11 (26.5%). To discover point mutations, 11 infants with negative MLPA results were sequenced. According to the results of the NGS, point mutations were identified in six boys (four single-nucleotide deletions and two single-nucleotide duplications), and five boys lacked mutations. Due to the high proportion of neuro-hereditary diseases in the general structure of neurological pathology, the profound disability of patients with progressive mental and physical disadaptation, as well as the generally fatal course of these incurable afflictions, molecular genetics research is of particular importance.

Reference intervals aid clinical decision-making for clinical chemistry values. Laboratory test results are compared to reference intervals to aid in the diagnosis, therapy, and monitoring decisions. Due to the differences in ethnicity, gender, age, and analytical methods, reference intervals (RIs) vary between populations. This study aimed to establish the reference values for renal function tests in targeted populations in Indonesia. This research was conducted with a cross-sectional observational analytic design. The research sample consisted of medical check-up data from health professionals at Dr. Mohammad Hoesin Hospital in Palembang, Indonesia. The Kolmogorov-Smirnov test was used to determine the normality of data distribution.  The RIs were computed using reference limits at the 2.5th and 97.5th percentiles (abnormal distribution) or ±two standard deviations (±2 SD) (normal distribution). The independent t-test (parametric) or Mann-Whitney test was used to compare the RIs of males and females (non-parametric). Males and females had a significant difference (P<0.001) regarding the values of uric acid, urea, and creatinine parameters, requiring the reference intervals to be separated. The following reference intervals were established: uric acid: 230,78-526,99 mol/L for males and 179,03-415.17 mol/L for females, urea: 2,22-4,99 mmol/L for males and 1,78-4,28 mmol/L for females, and creatinine: 61,01-106,99 mol/L for males and 40,67-77,81 mol/L for females. This study defined gender-specific RIs for three renal function test parameters for the adult population of Palembang, Indonesia. The deployment of population-specific RIs may facilitate better laboratory testing.

This study aimed to detect the levels of apurinic/apyrimidinic endonuclease 1 (APE1) gene expression and C-type lectin domain family 4 member M (CLEC4M) and their association with cisplatin chemotherapy in lung cancer patients. Overall, 105 individuals who attended the Al-Amal National Hospital for Cancer Management, Baghdad, Iraq, were enrolled in the study and divided into three equal groups. The groups included the patients newly diagnosed with lung cancer, cancer patients who received cisplatin, and the healthy control group. All study groups were subjected to the sampling of the venous blood for molecular analysis by real-time polymerase chain reaction (RT-PCR) to detect the APE1 gene and enzyme-linked immunosorbent assay (ELISA) for serological testing to measure the concentration of CLEC4M protein. Significantly, the values of both cancer groups were higher than those reported in the control group. The relative index revealed a significant difference in the mean fold change level of APE1 in the newly diagnosed group (3 fold) and cisplatin therapy patients group (2 fold), compared to the control group (P=0.005). No significant differences were detected between the two cancer groups in terms of fold change mean of expression, demographic characteristics, and cancer histological type. Regarding human CLEC4M protein level, cases receiving cisplatin (139.2±25.9) and newly diagnosed patients (331.0±38.1) had a highly significant difference with the control group (100.3±47.5, P<0.001). There was no significant difference between the concentration level of CLEC4M and all parameters in demographic characteristics and cancer histological type. This was the first study to demonstrate that higher expression levels of new APE1, CLEC4M, and glutathione, especially after chemotherapy, are beneficial as diagnostic and prognostic markers for resistance to platinum chemotherapy in Iraqi lung cancer patients.

This study aimed to examine the impact of Citrobacter freundii killed whole cell sonicated antigen (KWCSAg) alone and in combination with propolis nanoparticles on humoral immunoglobulin (IgG) and cellular immune responses of rats. The ELISA interleukin 4 (IL4) and IgG, delayed-type hypersensitivity (DTH) skin test, and phagocytosis activity tests were used in this study. In total, 45 rats were divided into five groups of 9 rats. The first group received a 1,000 μg\ml dose of KWCSAg-CF. The second group received an injection of 1,000 μg/ml of KWCSAg-CF antigen and 30 mg/ml of propolis AgNPs. The third group received an injection of 1,000 μg/ml of KWCSAg-CF antigen along with 10 mg/ml of propolis AgNPs. The fourth group was subjected to 30 mg/ml of propolis AgNPs. One ml of phosphate-buffered saline (pH 7.2) was injected into the fifth group (the negative control group). The rats received booster injections of the same antigens after 14 days. Blood was obtained from them to detect immunoglobulin and interleukin 4 (IL-4) on days 21, 28, 32, 46, 50, and 60 following the injection. The second group showed the most significant rise in IL-4 and IgG concentration, followed by the third group, the first group, and the fourth group, compared to the negative control group (fifth group). In all immunized groups, the DTH test results demonstrated an increase in the means of induration with significant differences (P˂0.05) of the concentrated antigen after 24 and 48 h, and subsequently a decrease after 72 h, compared to the negative control group. At 48 h after the concentrated antigen was indurated, the second group displayed the most significant increase in diameter.

One of the breast cancer subtypes, epidermal growth factor receptor 2 (HER2), accounts for 15% of all breast cancers and is characterized by aggressive behavior and a poor prognosis. For patients with HER2-positive breast cancer, trastuzumab, a monoclonal antibody that targets HER2 receptors, is prescribed in addition to chemotherapy to increase their chances of survival. However, the high expense of this treatment makes it impossible for patients in developing nations to easily afford it and undergo this biological therapy. Consequently, trastuzumab biosimilars have been launched as a substitute that offers comparable effectiveness at a reduced price. This study aimed to compare the biological activity and cardiac safety of reference trastuzumab with biosimilar trastuzumab by monitoring serum levels of the tumor biomarker CA15-3 and evaluating N-terminal pro-B-type natriuretic peptide (NT-proBNP) for the adverse cardiac effects of both treatments on HER2-positive breast cancer patients before and after six cycles of biological therapy. This prospective research was performed on 36 females with metastatic and early-stage HER2-positive breast cancer who visited the Oncology Department at Rizgary Hospital, Erbil, Iraq. The patients were within the age range of 30-80 years old. Eighteen individuals received reference trastuzumab, while the remaining 18 received both chemotherapy and biosimilar trastuzumab. Each patient had a data sheet that contained details from hospital-reserved files. In the Herceptin group, there was an insignificant difference in the median of CA15-3, while no significant difference was detected between the means of NT-proBNP before and after treatment. In the biosimilar group, there was a significant reduction in the median CA15-3 as well as a significant increase in the level of NT-proBNP before and after the treatment. Evaluation of the association of trastuzumab-induced cardiotoxicity during breast cancer treatment with different factors indicated that there might be an increased risk of cardiotoxicity after trastuzumab treatment.

Gastric cancer (GC) is one of the deadliest tumors due to its competence to invade and metastasize. The DNA repair gene (XRCC1), interleukin-8 (IL-8) gene, and B-cell lymphoma 2 (Bcl-2) gene play a crucial role in the development and progression of GC. This study aimed to evaluate the expression of these target genes in GC patients in the Kurdistan region of Iraq. Gastric cancer tissues were collected from 29 patients diagnosed with gastric adenocarcinoma that underwent gastric resection, and 21 tissue samples were obtained from healthy patients that underwent gastroscopy. The gastric tissues were collected in different hospitals in Erbil and Sulaymaniyah cities in the Kurdistan region of Iraq. Moreover, the data regarding Helicobacter pylori, age, gender, and stage of the disease were recorded and analyzed using GraphPad Prism. The gene expression levels of XRCC1, IL-8, and Bcl-2 from gastric tissue were studied by real-time quantitative polymerase chain reaction. The results showed that H. pylori infection was equally distributed among males and females in the tissues of gastric patients, while most of the H. pylori-negative patients were females. It is also found that gastric patients aged 30-60 years old are more commonly tested for the H. pylori test. Accordingly, in this study, patients diagnosed with gastric inflammation more often tested positive for H. pylori, while patients diagnosed with gastric cancer tested negative for this infection. Additionally, it was found that the target genes (XRCC1, IL-8, and Bcl-2) were significantly upregulated in GC patients, compared to the healthy group. Finally, the result revealed that XRCC1, IL-8, and Bcl-2 were upregulated in the Kurdish patients with GC, compared to the healthy control group. Targeting XRCC1, IL-8, and Bcl-2 genes can be an interesting field and promising strategy for cancer treatment.

Today, the human papillomavirus (HPV) L1 protein is the main target in the construction of prophylactic HPV vaccines. The production of virus-like particles (VLPs) that closely resemble the natural structure of the HPV16 virus and induce high levels of virus-neutralizing antibodies in animals and humans is facilitated by the expression of HPV16-L1 protein in eukaryotic cells. The Bac-to-Bac system has been previously used to produce high levels of recombinant proteins. In this study, we utilized this expression system to generate HPV16-L1 VLPs in Spodoptra frugipedra (Sf9) insect cells. The wild-type L1 gene of papillomavirus type 16 was selected from Gene Bank and placed in bacmid structure after codon optimization using pFast Bac vector. The recombinant baculovirus containing HPV-16/L1 gene was then provided using the Bac-to-Bac system. It should be mentioned that the vector was transfected into the Sf9 cell. The cells were then lysed and the expression of L1 protein was revealed by SDS-PAGE and confirmed by Western Blot. The L1 purification was performed through Ni-NTA chromatography. The VLP formation of papillomavirus L1 protein was visualized by transmission electron microscopy. The expressed recombinant L1 was ~60 KD on SDS-PAGE which was identified in western blot by a specific anti-L1 monoclonal antibody. The electron microscopy confirmed the assembly of VLPs. Results of this study showed that the production of this protein at the industrial level can be optimized using a baculovirus/Sf9 system. The characteristics and advantages of this system are promising and it is a suitable candidate for protein synthesis.

This study aimed to investigate the antibacterial and antifungal activities of selenium nanoparticles (SeNPs) and berberine (BBR) despite antibiotic resistance against Klebsiella pneumoniae and Candida albicans. Cells of K. pneumoniae and C. albicans were treated with solutions of different concentrations of each bare SeNPs, BBR, and BBR-loaded SeNPs (BLS) using the disk diffusion method. The results indicated that the activities of SeNPs, BBR, and BLS were statistically significant (P<0.05) when the concentration of all agents increased. Moreover, it was found that BLS had a statistically significant effect against K. pneumoniae and C. albicans, compared to SeNPs and BBR alone (P<0.05). The largest zones of inhibition of SeNPs were 14 and 16 mm toward K. pneumoniae and C. albicans, respectively, at the concentration of 20 Mml, compared to the concentrations of 10 and 15 Mml. Furthermore, BBR showed a maximum zone of inhibition at the concentration of 1,200 mg (15 mm for K. pneumoniae and 18 mm for C. albicans) and it was statistically significant in comparison with other concentrations of 400 and 800 mg. In addition, the BLS underwent a statistically significant increase (P<0.05) when the concentration increased and it registered a large zone of inhibition of 22 and 25 mm against K. pneumoniae and C. albicans, respectively, at 20 Mml of SeNPs: 1,200 mg BBR, compared to 10 Mml of SeNPs: 400 mg BBR and 15 Mml of SeNPs: 800 mg BBR. Based on the results of the current study, there was a statistically synergistic effect of BBR-loaded SeNPs, compared to that of BBR and Se nanoparticles, only in the case of both K. pneumoniae and C. albicans. This study is promising as a blueprint for the enhancement of weak antimicrobial agents and their return to their previous role as antibiotics.

In the transdermal drug delivery system, the drug is administered through the skin and attains a systemic effect. It is a drug administration route that includes drug transport to the epidermis and potentially dermal tissue of the skin for locally therapeutic effect, while an exceptionally significant drug division is transported in systemic blood circulation. This study aimed to formulate rasagiline mesylate (RM) as a transdermal microneedle (MN) delivery. The RM is an antiparkinson drug that can be classified as class III with low permeability and subjected to extensive first-pass metabolism. At first, it was formulated as nanoparticles using the chitosan polymer and ion gelation method. Afterward, the prepared nanoparticles were incorporated into a transdermal MN formulated by a polydimethylsiloxane template. The two-step casting process uses two polymer concentrations of polyvinyl alcohol and mixes them with other polymers in a 3:1 ratio (polyvinylpyrrolidone and chitosan) and glycerin as a plasticizer. The selected MN formula was MN4 with a promising shape, no bubbles, fine and well-formed sharp needles that passed the folding endurance test with 130 folding times before broken, drug content of 97±10.02%, and ex vivo permeation. The results showed a significant (P>0.05) permeability enhancement and increase of flux (160%), compared to the transdermal patch. The RS polymeric nanoparticles were successfully prepared and loaded within dissolving MNs of sufficient mechanical strength to penetrate the stratum corneum and enhance the amount permeated through it to induce the systemic effect transdermally.

Rasagiline is a selective and irreversible inhibitor of monoamine oxidase B (MAO-B) that is effective in the treatment of Parkinson’s disease (PD). It had antioxidant and anti-apoptotic activity in experimental models. Moreover, it has low permeability and its oral bioavailability is weak and highly variable due to extensive first-pass hepatic metabolism (35%). This study aimed to formulate rasagiline mesylate (RM) as a lipid-polymer hybrid nanoparticle in order to enhance its permeation and increase its chance to be absorbed by lymphatic circulation to avoid metabolism and control its release. Successful formulation (PCL-2) was reached by the nanoprecipitation method using polycaprolactone with RM in the organic phase and lecithin in the aqueous phase DSPE-PEG. The lipid:polymer ratio of 24% and DSPE: lecithin of 50% resulted in stable nanoparticles having a particle size of 132±4.58 nm, polydispersity index of 0.273±0.02, zeta potential of -25.6±3.3, entrapment efficiency of 46±3.9%, and drug loading of 51.93±6.5. Results showed that the diffusion was more effective on the release profile than the degradation and resulted in a Fickian diffusion mechanism.

Toxoplasmosis is one of the most widespread zoonotic diseases, especially in warm and humid areas, and affects all mammals, including humans and many herbivores and carnivores. The present study investigated the Toxoplasma gondii (T. gondii) parasite in tortoises for the first time in Iraq using PCR technology. A total of 28 tortoises/Testudo graeca (T. graeca) were collected between October 2018 and March 2019 from the study stations and then sent to the Animal House, which belongs to the Department of Biology, Faculty of Education, University of AL-Qadisiyah, Iraq, to perform the dissection. The body cavity was opened, and all organs were removed. The tortoises’ liver, heart, and brain were removed and kept at -20ºC until use. Afterward, the samples were subjected to DNA extraction. The Nested-PCR technique was implemented using two pairs of primers, and then the PCR products were analyzed using 1.5% agarose gel electrophoresis. The amplification of the gene during the first cycle indicated that 10 samples gave positive results with a total percentage of (11.9%), including five liver samples, three heart samples, and two brain samples (17.85%, 10.71%, and 7.14%, respectively). On the other hand, during the second cycle of the reaction, the amplification of the gene was obtained in seven samples (8.33%). The highest percentage of the presence of the gene was recorded in the tortoises’ liver (14.28%) and the lowest in their brain (3.57%). This study is among the first to investigate the molecular detection of T. gondii in wild tortoises (T. graeca) in Iraq. The findings imply that tortoises have a role in transmitting T. gondii and are believed to acquire infection by feeding on small invertebrate animals or plants contaminated with the oocysts of the parasite.

This study was conducted to confirm the phenotypic diagnosis of two Candida species, including Candida albicans (C. albicans) and Candida dubliniensis (C. dubliniensis). They were previously isolated in another study from cases of oral candidiasis using polymerase chain reaction and determining the nitrogenous base sequences of the 18 SrRNA product duplication using the NS1 and NS8 primers. The sequences of the multiple bases were analyzed using the Basic Local Alignment Search Tool program (BLAST), which proved that the two diagnosed Candida strains belong to two species, including C. albicans and C. dubliniensis, respectively. Additionally, the comparison of these sequences to the data available in the National Center for Biotechnology Information (NCBI) database showed that C. albicans strains in this study were 99% similar to the universal strains of C. albicans from Japan, Brazil, the United States, Germany, India, China, Pakistan, and Egypt. The C. dubliniensis strains in this study also had the highest genetic similarity rate of 99% to the C. dubliniensis strains isolated from the United States, Netherlands, France, and Germany. The study strains were recorded in the GenBank database with the sequence codes MZ574137 and MZ574410.1 for C. albicans and C. dubliniensis, respectively. The results of the 18 SrRNA region’s duplication also showed variations between C. albicans and C. dubliniensis, represented by the presence of three mutations of the first type and two mutations in the second type at different sequence sites.

Concurrent with an increase in the human population on the earth, more than ever, the creation of energy and maintenance of health is necessary, and nowadays, various sources of energy supply are being developed. The general global view in this regard is to provide protein and energy from available and cheap sources. Iran is no exception to this general rule, only in the field of ensuring the health of livestock resources every year, about 10 tons of peptone is needed for producing clostridial vaccines. Vermicomposting worms (Esienia fetida) with high protein percentages and rapid reproductions are a suitable source for peptone production. Based on this, the vaccine strain of Clostridium perfringens type D cultivated in two different media contain peptone produced from worms and meat peptone. The growth rate, epsilon toxin (ETX), and alpha toxin (CPA) of Cl. perfringens have been compared in two media. The results showed that the growth rate of bacteria in the worm peptone medium in 48 h was 22% higher than that of the meat peptone. Additionally, the activity of alpha toxin (phospholipase C) was in worm peptone 15% higher than meat peptone during 80 min of measurement. Regarding epsilon toxin lethality, all three mice of the N-worm peptone group died, while all three mice of the meat peptone group survived even 72 h after injection. The average survival time of mice in the N-worm peptone group was 1700 min. Therefore, we suggest the worms' protein is more suitable than industrial meat in peptone production for vicinal propose. To eliminate the need for hydrolyzed protein in the production of vaccines in the future, we suggest an increase in the fields of employment and the development of fertilizer and worm farms in Iran.

The severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, was first discovered in Wuhan, Hubei province, China. Cytokines play a critical role in COVID-19 infections through their inflammatory or anti-inflammatory activities. This study aimed to detect the diagnostic value of and the relationship between the interleukins under study, in addition to their relationship with demographic data in COVID-19 patients. Patients with a confirmed diagnosis of COVID-19 based on laboratory (PCR) results and the healthy control group were given their section of this investigation. The patient group had 120 COVID-19 patients, including 62 males and 58 females, while the control group consisted of 32 individuals (22 males and 10 females). The subdivision was then performed according to their vaccination status, chronic diseases, gender, and residence. Cytokine levels were detected using the ELISA technique. The immunological status of COVID-19 patients was determined by measuring interleukin (IL)-6, IL-25, and IL-35. During the research, it was found that IL-6 was highly significant in COVID-19 patients (0.001). However, its level was not significantly different (0.376) in patients regarding the type of chronic diseases, residence (0.353), and gender (0.574), but it was significantly different in vaccinated patients (0.029). It was also found that IL-6 is significantly correlated with IL-25 and IL-35. IL-25 was highly significant in COVID-19 patients (0.007), and there was a significant difference in its level in patients regarding the type of chronic disease (0.049). While there was no difference in terms of residence (0.421) and gender (0.681), corona vaccination showed a significant difference (0.047). IL-25 also had a significant correlation with IL-6 and IL-35. As for IL-35, it was significant in patients with COVID-19 (0.013) but not significantly different regarding chronic diseases (0.344), residence (0.877), or gender (0.800). However, it was significantly different in vaccinated patients, compared to the non-vaccinated ones. IL-35 was found to be significantly correlated with IL-25 and IL-6 (0.000). The examined interleukins increased in COVID-19 individuals. IL-6 remains an excellent marker for determining the immune state of patients with COVID-19. There were also strong correlations between the interleukins under study in COVID-19 patients. However, there was no relationship between age, residence, gender, and the concentration of studied cytokines. IL-25 increases significantly in COVID-19 patients suffering from chronic diseases. Therefore, it is more efficient in the follow-up of patients.

Adenoviral vectors (AdVs) are widely used as a gene delivery vehicle and vaccine design due to their genetic stability, transfer capacity of large genes, production at high titers, and remarkable efficacy of transduction. One of the most important applications of AdVs is in cancer immunotherapy. Tumor-associated antigens are overexpressed in cancer cells; however, they cannot induce immune responses sufficiently. Therefore, the immune system must be stimulated against these antigens to kill the cancer cells. This study described the construction steps of a recombinant AdV expressing human carcinoembryonic antigen (CEA) gene. Furthermore, in order to achieve a high titer of the virus, an efficient transfection was required. Three various transfection reagents were compared to achieve the best method of transfection. Carcinoembryonic antigen was cloned into the pAdV and transfected into the A293 cells using three different reagents, including polyethylenimine (PEI), calcium phosphate, and DMRIE-C. The PEI had the highest transfection efficiency, which was selected for the transfection of the recombinant plasmid. It has low toxicity for cells and is suitable for large-scale transfection. The virus produced in this study can be applied as a vaccine in cancer immunotherapy for stimulating the immune system against CEA-expressing tumors.

Chicken production is quickly rising due to the low associated costs and the capability of poultry to convert nutrients into biological protein along with chicken meat accounting for 30% of all animal protein eaten by humans. Despite advances in poultry production, parasitic illnesses in laying hens remain a problem. Farm birds reared in semi-intensive and free-range systems are more prone to parasite infections due to the absorption of polluted water and food from scavenging behaviors and waste droppings. In this study, the effects of Ascaridia galli infection on the immune response and liver function of two laying hen lines are compared, and their infection resistance is determined. In total, 50 laying hens at eight weeks of age were used (25 Lohmann brown-classic and 25 Lohmann lsl-lite), and each line was divided into two groups: an infected group (n=15), which was orally infected with a single dose of 500 A. galli embryonated eggs, and a control group (n=10), which was given normal saline. After four and eight weeks, blood was collected from the wing vein to assess the serum's AST, ALT, total protein, and IgY levels. The results demonstrated that the infected Lohmann brown-classic and Lohmann lsl-lite chickens presented significantly increased (P≤0.05) AST, ALT, and IgY, compared to the respective control group. Moreover, Lohmann brown-classic hens presented a significantly increased (P≤0.05) IgY concentration four weeks after infection, compared to Lohmann lsl-lite hens. From our results, it can be concluded that genetic variation plays a crucial role in the immune response against A. galli, where the Lohmann brown-classic line was found to be more resistant, compared to the Lohmann lsl-lite line.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mRNA vaccines, such as Pfizer-BioNTech, have demonstrated high efficacy; however, there is limited data on the duration of immune responses besides their relationships with age, gender, body mass index (BMI), and the presence of previous coronavirus disease-2019 (COVID-19) infection. This study aimed to evaluate SARS-COVID-19 Anti-Spike IgG levels after 30 days (one month) and 120 days (four months) of the 2nd dose of Pfizer-BioNTech vaccine given to medical students at Al-Iraqi University, Baghdad, Iraq. This study was performed after the obtainment of the acceptance and approval of the Medical College of Al-Iraqi University and the Iraqi Ministry of Health. Two groups of students were randomly picked up from the Medical College of Al-Iraqi University. They were completely vaccinated by administering two doses of Pfizer-BioNTech/0.5 ml for each dose. After taking their permission,  5 ml of their blood (one group after one month and the second group after four months of vaccination) was drawn in the Higher Education lab inside the Medical College of Al-Iraqi University. It took approximately four months to collect the samples (from October 2021 until February 2022). Following that, serological analysis was done for measuring the SARS-CoV-2 spike protein IgG by using Elabscience/SARS-CoV-2 spike protein IgG ELISA Kit (USA) (+ve <0.06) that was performed in the Higher Education lab of Medical College of Al-Iraqi University. Demographic data were also collected from participants, including age, gender, BMI, blood group, and the presence of previous COVID-19 infection. For statistical analysis, SPSS (version 26) and STATISTICA (version 12) were used to input, check, and analyze data. Standard approaches of frequencies and percentages were used for qualitative variables, while for quantitative variables, mean±standard deviation was used. A P-value of <0.05 was considered a significant plasma level of the SARS-COVID-19 Anti-Spike IgG. The study results showed that in group 1 (after one month of the 2nd dose), the male-female ratio was 62.2: 37.8, the mean age of the vaccinated students was 28.2000 years old, and the BMI was 25.5454 kg/m2 with 33.3% previously COVID-19 infected individuals. In group 2 (after four months of the 2nd dose), the male-female ratio was 44.4: 55.6, the mean age of the vaccinated students was 25.8444 years old, and the BMI was 24.7584 kg/m2 with 24.4% previously COVID-19 infected individuals. The plasma levels of SARS-COVID-19 Anti-Spike IgG after the 2nd dose of the Pfizer-BioNTech vaccine in group 1 (one month) and group 2 (four months) were statistically non-parametric. Once the independent two samples Mann-Whitney test was used, a significant difference (P<0.05) was observed in SARS-COVID-19 Anti-Spike IgG plasma levels after 30 days of the 2nd dose of the Pfizer-BioNTech vaccine administration, compared to the 120 days of administration. In conclusion, SARS-COVID-19 Anti-Spike IgG levels significantly increased in group 2 (four months after the 2nd dose of the Pfizer-BioNTech vaccine), compared to group 1 (one month after the 2nd dose of the Pfizer-BioNTech vaccine).

Feline infectious peritonitis (FIP) continues to be one of the most researched infectious diseases of cats. The diagnosis of FIP is challenging, and diverse techniques have been developed for its accurate diagnosis. However, they have some limitations. The present study was conducted to investigate the efficacy of specific modulation frequency (SMF), compared to other routine diagnostic methods for detecting feline coronavirus. Blood samples were collected from 30 diseased cats suspected of having FIP based on clinical signs. Electrophoresis, polymerase chain reaction (PCR), and SMF tests were performed for each sample. The sensitivity and specificity of each test, as well as the agreement between the tests and the gold standard (the combination of PCR, electrophoresis, and bioresonance results), were calculated using the Kappa coefficient method. The sensitivity and specificity of electrophoresis, PCR, and SMF for the diagnosis of FIP were 70.6%, 70.6%, 100%, and 100%, 72.7%, 81.8%, respectively. According to the findings of the present study, SMF is effective and safe in FIP diagnosis, which is a challenge in veterinary medicine diagnosis.

Burn injuries are the most frequent injuries in the world, with a death rate of 2.3-3.6%. Children and people of working age constitute 85-90% of the burn cases. Burn injury results in metabolic problems, a generalized inflammatory response, inefficient energy use, and other physiological alternations that may cause organ and system dysfunction and sepsis. Sepsis is mostly caused by multiple organ failures and has unique characteristics in burn injuries, which make it the most dangerous complication of burn injuries. This study aimed to investigate the correlation between sepsis in burn patients and the level of interleukin 8 (IL-8) in their serum. In total, 60 patients with burn injuries were included in this study. Blood samples were obtained from 60 burn patients and 30 healthy individuals as controls. The BacT/Alert and Vitek2 systems were used to identify the bacteria and determine their susceptibility to these bacteria. Moreover, enzyme-linked immunosorbent assay (ELISA) was used to determine IL-8 serum levels. Based on the results, elevated levels of IL-8 were observed in the serum of burn patients, compared to healthy individuals. Concentration of IL-8 was significantly higher in patients with sepsis, compared to healthy individuals without sepsis.

Egg drop syndrome (EDS) is a major viral infectious poultry disease with severe economic losses in laying hens. The disease is caused by an adenovirus and can be transmitted horizontally and vertically. This study investigated the EDS virus (EDSV) infection in duck embryo fibroblasts (DEF), specific pathogen-free (SPF) embryo fibroblasts, and SPF egg embryos using different methods. The results were compared to the virus culture in duck and SPF chicken eggs. Duck and chicken fibroblast cells were used as the primary cell culture in Dulbecco’s Modified Eagle Medium, and the low-pathogenic duck adenovirus was used to infect the ducks and SPF fibroblasts primary cell cultures, as well as the duck and SPF eggs. The titer of the virus was measured by hemagglutination assay, ECID50, plaque-forming unit, and TCID50 methods. The results revealed that EDSV could proliferate in the chorioallantoic membrane of DEF cells and duck eggs, compared to the chorioallantoic membrane of chicken embryo fibroblasts (CEF) and SPF chicken eggs. The findings showed that duck egg embryos and primary DEF cell lines are more appropriate for EDSV replication, compared to CEF and SPF chicken eggs. This suggests that the use of DEF culture for producing avian adenovirus EDS-76 is a suitable alternative for the embryonic egg culture.

Brucellosis is a zoonotic infection in livestock that induces a major public health concern in developing countries, including Iran. Despite the efforts of the Iranian veterinary organization (IVO) to control brucellosis, it is still prevalent in domestic animals. In this regard, the present study aimed to evaluate the efficiency of the control strategy used by the IVO in infected herds on serological, cultural, and molecular methods. For this purpose, blood specimens were sampled from a total of 8750 vaccinated dairy cattle in two Brucella-infected farms. These farms were recognized as positive for Brucella by a screening program. Sera were evaluated by the Rose Bengal Plate Test and Wright test analysis. Positive dairy cattle were slaughtered under IVO supervision. The remaining cattle were evaluated every 3 weeks and positive animals were slaughtered. This procedure continued until the remaining animals revealed three successive negative responses in serological tests. Several lymph nodes and milk samples were collected from 164 seropositive cattle and subjected to bacterial isolation and confirmation by Bruceladder-polymerase chain reaction. Brucella melitensis biovar 1 and RB51 vaccine strains were recovered from milk and lymph node samples, respectively. Shedding of B. melitensis in the milk of vaccinated cows is a serious problem resulting in the further spread of brucellosis. The policy of “test and slaughter” performed on infected dairy cattle farms showed their usefulness for the control of brucellosis outbreaks. For the uncontrolled spread of brucellosis in Iran, effective control of bovine brucellosis required several serological surveillances to identify infected herds, eradication of the reservoirs, and vaccination of young heifers with RB51.

Typhoid fever is one of the most commonly disseminated diseases and is considered to be linked to poor sanitation. It is responsible for 2-5% of all deaths, and its causative agent is Salmonella typhi. The current study aimed to investigate the antibacterial activity of prebiotics (inulin and starch) and probiotics against multidrug resistance of S. typhi bacterial isolates. Determination of the inhibitory effect of probiotics and prebiotics against S. typhi isolates was performed by agar well diffusion method and minimal inhibitory concentration. Body samples of all eligible patients were collected and cultured. Finally, 50 (25%) out of the total cultured samples were S. Typhi bacteria isolated from different samples. The bacteria were mainly found in blood, followed by stool and fluid (74%, 24%, and 2%, respectively). On differential medium, xylose lysine deoxycholate agar, the colonies appear red with black centers, while on MacConkey agar, the colonies appear smooth, pale, transparent, colorless, and raised. Regarding the inhibition zone values of bacteriocins of Lactobacillus from Yogurt against S. typhi in plate, significant differences were identified between the ones with and without prebiotic addition. Accordingly, the value of the inhibition zone for those without prebiotic addition (13.18±7.403) was significantly lower than that of cutoff values of 20 with a significant difference of -6.820 (t= -6.514, df: 49, P=0.000). Moreover, the inhibition effect of prebiotics (inulin and starch) against S. typhi at 37 °C for 24 h in part dish glucose as control, only the mean of inulin was found to be significantly lower than that of the cutoff value of 18 with the mean difference of -3.900 (t=-4.115, df: 49, P=0.000). Other prebiotics of glucose and starch in 24 h showed negative inhibition. Probiotics are live microorganisms that have beneficial host effects by enhancing microbial balance in the intestine, whereas prebiotics are indigestible food components having beneficial effects by enhancing the activity and growth of one or more colonic bacteria. Lactobacillus filtrates had considerable effects against the test S. typhi isolates.

Pregnancy toxemia (PT), also known as ketosis or twin lamb disease, is a group of in-sequence metabolic disorders usually observed in the last pregnancy period of ewes. Blood samples from 60 Awassi ewes were collected, including 50 ewes suffering from PT and 10 healthy ewes (2-8 years old) as a control group. All of them were in their final month of pregnancy from different regions of Salah Aldin Governorate, Iraq. The samples were collected between October 2021 and February 2022. Biochemical analysis of serum concentrations of all parameters was performed using the atomic absorption spectrophotometer, except for the beta-hydroxybutyrate and non-esterified fatty acids that were analyzed by enzyme-linked immunosorbent assay method. The results of the clinical criteria tests for temperature, respiration, and pulse showed nonsignificant differences (P<0.05) in the infected animals, compared to the healthy group. Clinical signs included depression, loss of appetite, weight loss, lying down, odor of ketogenic bodies through breathing, inability to walk, neurological signs, dental grinding, jaundice, blindness, bloat, dystocia, animal death, and fetal death. Based on the results of the biochemical parameters tests of the blood, a significant increase (P<0. 05) was observed in the parameters of the results of beta-hydroxybutyrate, non-esterified fatty acids, triglycerides, total bilirubin, and liver enzymes (ALT, AST, ALP, and GGT) in the animals affected by PT, compared to the control group. However, a significant decrease (P<0.05) was observed in the parameters of glucose, cholesterol, total protein, albumin, and globulin in the affected animals, compared to the healthy group. Concerning the association between disease and oxidative stress criteria, the infected animals showed a substantial (P<0.05) increase in malondialdehyde concentration and a significant (P<0.05) drop in glutathione and superoxide dismutase levels.

Liver function tests are frequently used to screen liver function, monitor therapy, and determine the severity of liver problems. The present study aimed to assess the consistency of the results of the liver function parameters between the two analyzers, Architect c8000 and Cobas C501. This laboratory-based analytical observational study was conducted in a cross-sectional manner. Sample collection was performed through a consecutive sampling procedure from June to December 2019 in the Clinical Pathology Laboratory, Dr. Mohammad Hoesin General Hospital, Palembang, South Sumatra, Indonesia. The research sample consisted of the liver function examination results of patients, carried out using the Architect c8000 and Roche Cobas c501 chemistry analyzers. Serum albumin, alanine transaminase, aspartate aminotransferase, and total protein were the studied variables. The Spearman, Mann-Whitney, and Bland-Altman tests were used to evaluate the comparison test. In total, 100 blood samples were obtained in this study. The results revealed a highly significant correlation (r>0.90, P=0<001) among the four liver function parameters. The results of the liver function parameters inspected by the two analyzers did not differ significantly (P>0.05). In addition, there was a solid agreement on all parameters, with a near-perfect level (concordance correlation coefficient>0.90) and more than 95% of data points falling within the acceptable range. The Architect c8000 and Cobas c501 analyzers produced similar results for liver function tests; hence, these devices can be used interchangeably. 

Infertility is defined as the inability of couples to conceive after 1 year of regular unprotected intercourse, which affects 10-15% of couples. The present study aimed to investigate the influence of Interleukin-17 (IL-17) and growth differentiation factor 9 (GDF9) on three groups of infertile males, including control, azoospermia, and oligozoospermia. In total, this study was performed on 93 participants, consisting of 18, 65, and 10 subjects in the Azoospermia, oligozoospermia, and control groups, respectively. The mean plasma levels of IL-17 in the azoospermia and oligozoospermia groups were 21.317±3.605 and 15.101±2.416 ng/l, respectively, which were significantly higher than that in the control group (5.392±1.731 ng/l). Furthermore, the mean plasma levels of GDF9 in the azoospermia and oligozoospermia groups were 3.299±1.051 and 6.2603±2.621 ng/l, respectively, which was significantly higher than that in the control group (12.807±2.170 ng/l). One-way analysis of variance and least significant difference post-hoc test were performed to assess significant differences among means. R-squared measures how well the linear regression model fits the data. It can be interpreted as the proportion of variance of the outcome Y explained by the linear regression model. R-squared is a number between 0 and 1. In non-obstructive forms of severe oligozoospermia and azoospermia, like the case in the current study, intracytoplasmic sperm injection is suggested by using testicular biopsy for spermatozoa extraction, if viable spermatozoa are present.

The present research aimed to study the polymorphisms of the chicken insulin-like growth factor 2 (IGF2) in two commercial broiler breeds (Cobb 500 and Hubbard F-15). In total, 300 avian blood samples were obtained. The genomic DNA was isolated using a fast salt-extraction technique. Moreover, polymerase chain reaction (PCR) was used to amplify 1146 bp fragments of the gene. The amplified fragments were subjected to restriction enzyme digestion using the HinfI endonuclease enzyme, and the digested products were separated on a 2% agarose gel. The findings indicated that there were two alleles T and C for the target locus, with frequencies of 73.3% and 26.7%, respectively. Three distinct genotype variations, TT, TC, and CC, were found, with genotype frequencies of 59.1%, 28.4%, and 12.5%, respectively. A test based on actual and anticipated frequencies of various genotypic variances of the IGF2 gene revealed that the divergence from Hardy-Weinberg equilibrium was not significant (P≤0.01) in commercial broiler breeds (Cobb 500 and Hubbard F-15) chickens. In addition, it was found that birds with genotype TC had a greater body mass at 8 weeks of age, compared to those with genotypes TT and CC. It was determined that the IGF2 gene exhibited a significant degree of variability and might be regarded as a possible genetic marker in selection and breeding programs for poultry.

Clostridial enteric diseases, called enterotoxemia, are caused by Clostridium perfringens toxinotypes in sheep and other ruminants. This study aimed to describe the molecular characterization of C. perfringens isolates in diarrhoeic sheep (Ovis aries) flocks in the southeast of Iran. Fecal/intestinal samples were collected from diarrhoeic (n=116), dead (n= 13), and healthy (n=63) sheep over four years (2016-2020) and subjected to bacteriological and molecular examinations. The C. perfringens isolates were typed by polymerase chain reaction targeting genes, namely 16SrRNA, CPA, CPB, ETX, IAP, CPE, and NetB. The overall prevalence of C. perfringens was 28.6% among the studied sheep, and there was a significant relationship between its isolation rate and diarrhea (P<0.001). The C. perfringens isolation rate also decreased with animal age (P=0.012) and was significantly higher in late winter and spring (P=0.000). The most prevalent toxinotypes were types A (52.4%), D (22.2%), and F (18.5%), in that order. Moreover, C, G, and B types were found in 4.2%, 1.6%, and 1.1% of the isolates, respectively, and no type E was detected. The CPE gene was detected in 32.3% of all isolates, and the diarrhoeic sheep were most likely to yield CPE+ strains of C. perfringens (93.1%). These findings highlight the importance of CPE+ strains of C. perfringens in sheep enteritis and suggest that the high presence of type F needs to be considered in new clostridial vaccines containing this toxinotype. It is noteworthy that the present study reported the isolation of C. perfringens type F, type G, and the CPE+ strains of type B from diarrhoeic sheep for the first time.