Evaluation of sperm’s chromatin quality with acridine orange test, chromomycin A3 and aniline blue staining in couples with unexplained recurrent abortion

Objective

To evaluate the sperm’s chromatin quality in couples with spontaneous recurrent abortion.

Methods

Thirty couples with spontaneous recurrent abortion (case group) and 30 fertile couples (control group) referring to Zeinabieh Gynecology clinic of Shiraz were included. Semen samples were collected for each participant and were used for standard semen analysis and sperm nuclear maturity tests including Chromomycin A3 (CMA3), Aniline Blue (AB) staining and Acridine Orange (AO) test (by light microscopy).

Result

Patients in case group had significantly higher percentage of CMA3 (p?<?0.001) and AB (p?<?0.001) positive spermatozoa compared to controls. However AO results did not differ significantly between groups (p?=?0.656). Sperm morphology and progressive motility were negatively correlated with CMA3 (p?=?0.001 and p?=?0.043) and AB (p?=?0.015 and p?=?0.031) respectively.

Conclusion

Evaluation of the sperm’s quality via CMA3 and AB staining could be considered as one of the complementary tests of semen analysis for assessment of male factor in couples with spontaneous recurrent abortion.     

Evaluation of blood supply operation and infectious disease markers in blood donors during the Egyptian revolution

BACKGROUND: The Egyptian revolution took place on January 25, 2011. Millions of protesters demanded the overthrow of the Egyptian president's regime. Many people suffered from life‐threatening injuries after violent clashes between police and protesters. STUDY DESIGN AND METHODS: The overall management of the blood bank operation at Cairo University Hospital was described, in an attempt to evaluate blood safety and establish a standard effective plan to manage blood supply during crisis. RESULTS: Three days after the uprising, thousands of Egyptians rushed to the hospital to alleviate the blood shortage. A total of 3425 units were collected in 3 days and thousands of donors were turned away. An error delayed processing of 1000 units and they were used as stored whole blood. Apheresis platelets were donated by protesters who were particularly motivated to donate for two victims with liver injury. The usual positive rate of hepatitis C virus (HCV) antibody in Egyptian donors is 3.8%. However, the positive rate of HCV markers in the collected units was only 1.6%. The mean age of donors during the revolution was 31.7 ± 10.4 years while the usual mean age of donors is 39.2 ± 8.5 years. Operating theaters were used only for emergencies. A blood surplus developed that met the hospital needs for 1 month. CONCLUSION: Revolution resulted in an influx of first‐time donors with a relatively low positive rate of HCV antibody. To be prepared for disasters, a systematic approach to spread donors evenly on a daily basis is needed.     

Role of cervical spine radiography in the initial evaluation of stable high-energy blunt trauma patients

Background

Detecting cervical spine injuries in trauma patients is essential because undetected injuries in the this area may result in severe neurological disability and probably quadriplegia. Thus, radiography of the cervical spine is considered mandatory in the initial evaluation of trauma patients according to Advanced Trauma Life Support. This approach results in many unnecessary normal radiographs. Therefore, we performed this study to determine the role of routine cervical radiography in the initial evaluation of stable high-energy blunt trauma patients.

Methods

This was a prospective cross-sectional study including all hemodynamically stable high-energy blunt trauma patients with negative cervical physical examinations referred to our trauma center during a 5-month period (May to September 2010). Cervical radiographs, computed tomography (CT) scanning and magnetic resonance imaging were performed and reviewed for abnormalities.

Results

During the study period, 1,679 high-energy blunt trauma patients were referred to our center, of which 400 were hemodynamically stable and had negative cervical physical examinations. Cervical radiographs were found to be normal in all patients.

Conclusion

Cross-table cervical spine radiographs can be limited to those high-energy blunt trauma patients who have a positive cervical physical examination or those in whom the physical examination is not revealing. These radiographs also have low value for detecting occult cervical spine fractures, and CT imaging is considered the modality of choice in these cases.     

Development and evaluation of a multiplex PCR for rapid detection and differentiation of Mycobacterium tuberculosis complex members from non-tuberculous mycobacteria

Most mycobacterial infections are still caused by Mycobacterium tuberculosis complex (MTC) strains; however, infections by non-tuberculous mycobacteria (NTM) are increasing, particularly among immunocompromised patients. Conventional species-specific identification and proper patient management are delayed due to the slow-growing nature of mycobacteria. We have developed a multiplex PCR (mPCR) targeting the oxyR-ahpC intergenic region and rpoB gene for direct detection and differentiation of clinical isolates as MTC or NTM in primary culture. Two amplicons of 473 bp and 235 bp from MTC members and a single amplicon of 136 bp from NTM are expected. The mPCR was developed using several mycobacterial species and was evaluated by testing extracted DNA from liquid cultures, flagged as positive for bacterial growth, of 100 consecutive mycobacterial isolates. The results were validated by DNA sequencing of the species-specific 16S-23S internal transcribed spacer (ITS) region. The mPCR with template DNA from reference Mycobacterium spp. yielded the expected amplicons. When 100 consecutive clinical isolates of Mycobacterium spp. were tested, 92 strains yielded MTC member-specific amplicons, and DNA sequences from 10 randomly selected isolates matched completely with the ITS sequence from M. tuberculosis. Eight isolates were identified as NTM, and DNA sequencing of the ITS region confirmed the NTM status of each of these isolates. The mPCR developed in this study allowed rapid detection and differentiation of primary cultures as MTC or NTM, thus helping in timely institution of specific therapy.     

Prevalence of Candida dubliniensis among cancer patients in Kuwait: a 5‐year retrospective study

Despite close genetic and phenotypic relationship of Candida dubliniensis with Candida albicans, its role in human disease is mostly restricted to oral colonisation, particularly among HIV‐infected patients. The prevalence of C. dubliniensis in association with other disease conditions has been infrequently reported. In this study, we present data on the prevalence of C. dubliniensis among yeast species isolated from cancer patients over a 5‐year period. A total of 1445 yeast isolates recovered from respiratory specimens, blood, urine and oral swabs were analysed. Candida dubliniensis isolates were provisionally identified by phenotypic methods and their identity was further confirmed by species‐specific amplification and/or sequencing of internally transcribed spacer region of rDNA. Antifungal susceptibility for fluconazole was determined by Etest. The number of isolates identified as C. dubliniensis, C. albicans and other yeast species were 71 (4.9%), 862 (59.6%) and 512 (35%) respectively. All the C. dubliniensis isolates originated from respiratory (5.9%) or oral (3.2%) specimens with an overall prevalence of 4.9%, and were found to be susceptible to fluconazole. The isolation of C. dubliniensis from respiratory or oral specimens and not from blood or urine specimens suggests that this species has preference to colonise these sites of human body.     

Modelling the role of microbial p-cresol in colorectal genotoxicity

Background: A greater understanding of mechanisms explaining the interactions between diet and the gut microbiota in colorectal cancer is desirable. Genotoxic microbial metabolites present in the colon may be implicated in carcinogenesis and potentially influenced by diet.

Aims: We hypothesised that microbial p-cresol is a colonic genotoxin and set out to model potential exposures in the colon and the effects of these exposures on colonic cells.

Methods: Batch culture fermentations with human faecal inoculate were used to determine the synthesis of p-cresol and other metabolites in response to various substrates. The fermentation supernatants were evaluated for genotoxicity and the independent effects of p-cresol on colonic cells were studied in vitro.

Results: In batch culture fermentation, supplementary protein increased the synthesis of phenols, indoles and p-cresol, whereas supplementary fructoligosaccharide (FOS) increased the synthesis of short chain fatty acids. The p-cresol was the greatest predictor of genotoxicity against colonocytes in the fermentation supernatants. Spiking fermentation supernatants with exogenous p-cresol further increased DNA damage, and independently p-cresol induced DNA damage in a dose-dependent manner against HT29 and Caco-2 cells and influenced cell cycle kinetics.

Conclusions: In the colon p-cresol may reach physiologically significant concentrations which contribute to genotoxic exposures in the intestinal lumen, p-cresol production may be attenuated by substrate, and therefore diet, making it a potential modifiable biomarker of genotoxicity in the colon.