Advanced Biomedical Research
Volume:9 Issue: 5, May 2019

Tumor-targeting peptides are attracting subjects in cancer therapy. These peptides, which are widely studied, deliver therapeutic agents to the specific sites of tumors. In this study, we produced a new form of recombinant listeriolysin O (LLO) with genetically fused Anti-HER2/neu peptide (AHNP) sequence adding to its C-terminal end. The aim of the study was to engineer this pore-forming toxin to make it much more specific to tumor cells. Materials and Method and

Two forms of the toxin (with and without peptide) were subcloned into a bacterial expression plasmid. Subcloning was performed using a polymerase chain reaction (PCR) product as a megaprimer in a quick-change PCR to introduce the whole insert gene into the expression plasmid. After expression of two recombinant forms of LLO in BL21 DE3 cells, purification was performed using Ni-NTA affinity column. MDA-MB-231 and MCF-7 cell lines (as negative and positive controls, respectively) were treated with both LLO toxins to evaluate their cytotoxicity and specificity. The IC50 of LLO on MDA-MB-231 and MCF-7 cells was 21 and 5 ng/ml, respectively. In addition, IC50 for the fusion AHNP-LLO toxin was 140 and 60 ng/ml, respectively. It was found that the cytotoxicity of the new engineered AHNP-LLO toxin has decreased by about 9x compared to the wild-type toxin and the specificity of the AHNP-LLO toxin has been also reduced.

Results show that the C-terminal of the LLO should not be modified and it seems that N-terminal of the toxin should be preferred for engineering and adding peptide modules.

Although undescended testis (UDT) is the most prevalent malformation in male neonates, the best mode of UDT treatment remains controversial. This study aimed to compare trans-scrotal incision approach with traditional trans-scrotal incision orchiopexy in children suffering from cryptorchidism.

This single-blind randomized clinical trial was done on 100 children with UDT who needed surgery. The participants were alternately undergoing trans-scrotal incision orchiopexy (Group I) and traditional inguinal incision orchiopexy (Group II). The success rate and incidence of postoperative complications were evaluated 1 week and 1 month and 6 months after the operation in the two groups.

Both the groups were similar in baseline characteristics including age and laterality (P > 0.05). There was no significant difference between the two groups in terms of the incidence of wound infection, testicular atrophy, testicular hypotrophy, and relapse (P > 0.05). In addition, the success rates were 98% in Group I and 94% in Group II (P > 0.05).

Both surgical methods have a high success rate, and there is no significant difference in the incidence of complications; however, in terms of beauty, satisfaction, and shortening the duration of surgery and the duration of hospitalization, trans-scrotal approach was more successful than the traditional method.

Mayer–Rokitansky–Küster–Hauser (MRKH) Syndrome is a female reproductive system disorder. It is characterized by a defect in the Müllerian ducts development, and it causes the absence of the uterus in variable degrees in upper vaginal hypoplasia. In addition, it is often associated with the unilateral renal dysplasia. Müllerian agenesis affects 1 in 4500 newborn girls and is considered as a sporadic anomaly. Women with MRKH Syndrome have a normal female chromosome pattern 46, XX with normal ovarian function. The presence of bilateral kidney agenesis with a pelvic pancake-shaped kidney is a rare condition, and a few cases have been reported in medical journals. This case study focuses on a case of MRKH Syndrome with bilateral renal agenesis and a pancake-shaped kidney.

Bladder outlet obstruction (BOO) is a relatively infrequent urologic condition in women, but can cause bothersome symptoms. In this article, transurethral incisions of the bladder neck (TUIBN) and urethra in the treatment of anatomical BOO were assessed.

A total of 23 women who referred with chronic lower urinary tract symptoms, urinary retention, and difficulty in micturition were assessed. Diagnose was made on the basis of urethrocystoscopy, voiding cystourethrography, and urodynamic studies. All patients underwent transurethral incisions of bladder neck contracture or site of observed urethral stenosis at 3- and 9-o'clock positions. Clinical improvements and complications were assessed by follow-up examination and International Prostate Symptom Score (IPSS) and quality of life (QOL) scoring before and after intervention.

All patients were followed for at least 6 months after intervention. Follow-up data were available for 19 (90.5%), 13 (61.9%), and 7 (33.3%) of patients at 12, 24, and 48 months' follow-up, respectively. During the follow-up period, the mean IPSS and QOL significantly changed from 26.84 to 10.74 and 4.76 to 2.32, respectively (P < 0.001). Approximately 66.7% had satisfactory outcomes, 47.7% patients after first, and 19.1% after second TUI. About9.5% patients developed new onset of stress urinary incontinence, one of them had indications of surgical intervention, and thus, symptoms were relieved by mid-urethral sling.

Transurethral incision of bladder neck and urethra seems to be effective in relieving urinary symptoms of anatomical BOO in women. Complications may rarely occur and can be fully managed.

The development of protective vaccines for Brucella spp. has been hampered by the difficulty in transformation of Brucella cells with foreign DNA for genetic manipulation. It seems that the formation of Brucella spheroplasts would increase the efficiency of transformation. The aim of this study was to devise an efficient method for the transformation of Brucella spp.

At first, spheroplast of Brucella was prepared by glycine and ampicillin induction and transformed using optimized protocols of CaCl2, electroporation, and lipofection methods. Then, the efficacy of transformation was compared between the three-mentioned methods.

Ampicillin-induced spheroplasts from early-log phase culture of brucella when incubated in a medium-containing 0.2 M sucrose during cell recovery had higher transformation efficiency in three different methods. Comparison of the transformation efficiency of Brucella abortus RB51 using the CaCl2, lipofection, and electroporation methods revealed that the transformation efficiency with the lipofection method was significantly higher than with other two methods (P < 0.05).

Lipofection method by lipofectamine 2000 on ampicillin-induced spheroplasts can be a suitable approach for Brucella transformation.