International Journal of Medical Laboratory
Volume:9 Issue: 4, Nov 2022

Hypertyrosinemia type 3 (HT3) is an inherited error in tyrosine metabolism caused by a mutation in the 4-hydroxyphenylpyruvate dioxygenase (HPD) gene. Here we report a one and half-year-old girl infant who was diagnosed based on increased serum tyrosine levels and increased urinary excretion of p-hydroxyphenyl derivatives.

The proband was one and half-year-old Iranian girl who was diagnosed based on increased serum tyrosine levels and increased urinary excretion of p-hydroxyphenyl derivatives. In this study, we used Whole-Exome Sequencing to identify the genetic reason for the disease and the funded mutation confirmed by Sanger sequencing.
Results and

Through whole-exome sequencing screening of heterozygotes c.413C>T (p.T138M) and c.75G.A (p.W25Ter) in the HPD gene and genetically confirmed by Sanger sequencing. There were heterozygous conditions c.413C>T (p.T138M) and c.75G.A (p.W25Ter) in father and mother respectively. This mutation in her parents was also confirmed by Sanger sequencing.

It has been proven that human mesenchymal stem cells (MSCs) conditioned medium (hMSCs-CM) can influence human embryonic stem cells (hESCs) chondrogenic differentiation. In this study, we hypothesized that conditioned medium (CM) from hESCs-derived MSCs in a sequential 3D-2D culture system could facilitate the induction of chondrogenesis in hESCs.

CM was collected from Yazd2 (hESCs; 46, XY) derived MSCs confluent cultures and stored at -20 °C. Yazd4 hESC line (46, XX) was induced for differentiation using EB formation as 3D culture into SD (spontaneously differentiation) and CM groups (differentiation using conditioned medium) for four days. Cell culture continued in a 2D (monolayer) culture system for both groups till day 14. Ultimately, chondrogenic differentiation was assessed by Alcian blue and masson′s trichrome staining at 4 and 14 days of differentiation, and quantitative real-time polymerase chain reaction (PCR) for NANOG, MEOX1, SOX5, SOX6, SOX9, ACAN, COL2A1 and RUNX2 genes for SD and CM groups on days 0 and 4.

  The gene expression profile for chondrogenic genes in the CM group was significantly more than the SD group (p< 0.05). Furthermore, chemical staining assessment illustrated a significant GAG and collagen II difference between the CM and SD groups at days 4 and 14 (p< 0.05).

Our findings would pave the way for creating an in vitro human chondrogenesis model for further studies in the developmental biology of articular cartilage tissue, which lend itself to cell-based therapy to cure joint diseases such as osteoarthritis.

Emerging antimicrobial resistance is one of the major public health threats worldwide. It can result in increased morbidity and mortality rates along with increased treatment costs and hospitalization stays. Methicillin-resistant Staphylococcus aureus (MRSA) has become a global challenge. It is a major cause of nosocomial and community-acquired infections. Its prevalence varies with country and within hospitals within a country. The current study estimates the prevalence of MRSA.

A total of 162 Staphylococcus aureus strains were isolated from various clinical specimens, and antibiotic susceptibility tests and identification by cefoxitin disc (30 μg) were performed as per Clinical and Laboratory Standards Institute guidelines.

Among 162 Staphylococcus aureus 92 isolates were found to be MRSA. The highest rate of resistance was detected for penicillin (100), followed by erythromycin (97%), clindamycin (57%), tetracycline (13%), sulfamethoxazole-trimethoprim (SXT) (55%), ciprofloxacin (23%), and gentamicin (2%). All of the isolates were susceptible to linezolid and vancomycin.

Treatment of multi-drug resistant MRSA is still problematic because of the limited choice of antibiotic. This analysis will assist clinicians to choose an appropriate course of action for MRSA infections.

Recurrent pregnancy loss (RPL) is one of the important complications of pregnancy. Only 50% of cases have a known cause of RPL. One of the causes of RPL is Waddlia Chondrophila (W. Chondrophila), but no information is available regarding its in Iran. This study aimed to develop a Taqman Real-Time polymerase chain reaction assay to detect recA Gene of W. Chondrophilain biological samples from women with RPL in Yazd city.

Clinical samples of women with a history of RPL, normal childbirth, and standard strain WSU 86–1044 were provided. Primers and probes of W. Chondrophila were designed. Positive control was provided based on the PUC57 vector. Technical performance was examined using a control plasmid.

Analytical sensitivity was 5 ng/µl of control plasmid DNA. No cross-amplification was observed when testing other pathogens that can detect in human vaginitis. The Taqman Real-Time PCR assay exhibited good reproducibility. This Real-Time PCR was then applied to 100 vaginal swabs. No samples were revealed to be Waddlia positive.

This new TaqMan Real-Time PCR assay represents a diagnostic tool that may be used to further study the prevalence of W. Chondrophila infection in clinical specimens.

Toxocariasis is a zoonotic parasitic disease with worldwide distribution caused by the larval stage of ascaridoid nematodes of dogs (Toxocara canis) and cats (Toxocara cati). The study was accomplished to determine the sequence variation in ITS2 and COX1 genes within isolates of Toxocara canis.

Two hundred Stool samples were collected randomly from dogs in public parks and streets from different regions of Zabol. Thirty samples containing eggs were isolated from the feces using Formalin ether 10% and centrifugal flotation. Genomic DNA was extracted, and COX1 and ITS2 were amplified by PCR-RFLP and sequenced. Sequence data were aligned using the BioEdit software and BLAST program and compared with published sequences in GenBank. The phylogenetic relationship between isolates of T. canis from Zabol city with other regions based on sequences obtained from COX1 and ITS2 genes and using MEGA7.0 software was investigated.

For all samples, amplicons of about 388 and 422 base pairs were produced by PCR for ITS2 and COX1, respectively. Drawing the phylogenic tree of ITS2 and COX1 sequences of isolates of T. canis  showed that the identified genotypes are not only different from each other but also from other parts of the world.

Our result showed that genetic variation among isolates of T. canis from Zabol is very low. For a deeper understanding of genetic diversity among populations of Toxocara, it is recommended to analyze more isolates from various geographical areas and variable genetic markers.

Polycystic ovary syndrome (PCOS) has a largely unknown etiology and is a common heterogeneous endocrine disorder in women of reproductive age, characterized by polycystic ovaries, hyperandrogenism, insulin resistance, and chronic anovulation. MicroRNAs (miRNAs) are small, noncoding RNA sequences that negatively regulate gene expression at the post-transcriptional level. miRNA-103 has been associated with metabolic disorders such as obesity and diabetes, which are also associated with PCOS. This study aims to describe the different expressions of circulating miR-103 in the plasma of PCOS women.

This case-control study includes 25 women with PCOS and 25 healthy women. Plasma total RNA was isolated and, after polyadenylation, converted to total cDNA. Then, the expression level of microRNA-103 was analyzed by quantitative reverse transcription-polymerase chain reaction, and SNORD miRNA was used as the reference gene.

Our results indicate that the expression level of miRNA-103 significantly decreased in the PCOS group compared to healthy women, which can be related to the etiology and progression of PCOS (p < 0.05).

Our findings recommend further studies in the larger statistical population and more accurate techniques to confirm microRNA-103 as a noninvasive diagnosis biomarker.

One of the most important genes involved in Alzheimer's disease (AD) is the presenilin2 (PSEN2) gene, which is one of the main constituents of the gamma-secretase complex. Mutations in this gene promote the formation of amyloid plaques resulting in AD. The study aimed to evaluate the mutation variant in exon 6 of the PSEN2 gene in patients with Late-Onset AD (LOAD). Due to the important role of the PSEN2 gene in the formation of beta-amyloid aggregates and the investigation involves an association between PSEN2 mutations and their pathogenicity in LOAD progression, we presented this exon as a more efficient alternative.

The thirty patients with LOAD and 16 healthy subjects as a control group were involved in this experimental study. DNA was extracted from blood samples and purified. The desired gene fragment was propagated using polymerase chain reaction and the products were electrophoresed and the results were analyzed.

A novel mutation was found in PSEN2 IVS6 + 30 G → C at the intron region of exon 6 in 20 cases of patients suffering from LOAD and 12 subjects in control cohort. In this mutation a guanine base was substituted by cytosine base which this position was 30 nucleotides separated from coding region.

The novel mutation was identified in both studied groups. These findings reveal no relationship between PSEN2 mutation and pathogenicity of LOAD disease. However, further studies are required to find the role of PSEN2 mutation in LOAD progression.

Human leukocyte antigen (HLA) molecules play a substantial role in T Lymphocyte-mediated adaptive immune response. Down-regulation of HLA expression may help the tumor to escape from immune surveillance. This study aimed to evaluate the correlation between HLA-G and HLA-E tissue distribution and the degree of tumor malignancy in human breast tumors.

This cross-sectional study was conducted
on tissue samples of 145 patients with breast cancer. The distribution of HLA-G and HLA-E expressions were measured by the immunohistochemistry method.

Among 145 patients, 51% of tumors did not express HLA-E, and +1 positive was seen in 36.6 % of patients. +2 positive in 8.3% and +3 positive in 4.1% of patients. Moreover, 79.3% of tumors did not express HLA-G, 17.9% expressed +1 positive, 2.8% expressed +2 positive, and no patients expressed +3 positive. Generally, 20.7% and 49% of patients showed expression of HLA-G and HLA-E, respectively. A significant correlation was seen between the grade of disease and expression of HLA-E (p = 0.03) and HLA-G (p = 0.001). Moreover, a significant correlation was seen between the simultaneous expression of HLA-G and HLA-E with grade (p = 0.03, r = 0.176).

Significant correlation was seen between the distribution of HLA-G and HLA-E expression with a degree of malignancy. Therefore, these biomarkers’ expression may contribute to the prognosis and progression of breast cancer.