Moreshwar R. Shende

Genetic Risk of Azoospermia Factor (AZF) Microdeletions in Idiopathic Cases of Azoospermia and Oligozoospermia in Central Indian Population

BACKGROUND Genetic factors cause about 15% of male infertility. Azoospermia factors (AZFa, AZFb, and AZFc) present on Yq are most important for spermatogenesis. We have made an attempt to evaluate the frequencies of microdeletions of AZFa, AZFb, AZFc in idiopathic cases of azoospermia and oligozoospermia from central Indian population. MATERIALS AND METHODS We have analyzed a total of 156 subjects (95 oligozoospermia and 61 azoospermia) & 50 control subjects. DNA samples were analyzed for microdeletions of Y chromosome by PCR-screening of 18 sequences-tagged-site (STS) markers from different region of the AZF on Yq and SRY on Yp. RESULTS Out of 156 cases analyzed, 13 (8.33%) subjects (8 azoospermia and 5 oligozoospermia) showed partial deletion of AZF regions, of which deletion in AZFc region was the most common (84.6%) followed by AZFb (15.4%) and AZFa (15.4%). The sites and sizes of deletions varied among patients. Histological study of the testicular tissue of the available subjects, who showed microdeletions of Y chromosome, showed spermatogenic arrest at different stages. The frequency of Y chromosome microdeletion in our subjects was 8.33%. CONCLUSION Some Indian studies reported low frequencies of microdeletions than that of our result. We suggest that the frequency of deletions may be affected by the involvement of different genetic factors, ethnic population and different geographical regions. PCR based Y chromosome screening for microdeletions will be useful and great help to infertility clinics for genetic counselling and assisted reproduction.

SXP–RAL Family Filarial Protein, rWbL2, Prevents Development of DSS-Induced Acute Ulcerative Colitis

Helminthic infections lead to the release of various molecules which play an important role in modulation of the host immune system. Such filarial proteins with immunomodulatory potential can be used for therapeutic purpose in inflammatory and immune mediated diseases. In the present study, we have explored the prophylactic effect of filarial SXP–RAL family protein of Wuchereria bancrofti i.e. rWbL2 protein in DSS induced inflammatory ulcerative colitis in a mouse model. Prior treatment of rWbL2, followed by induction of colitis, showed significantly reduced disease severity as indicated by the decreased disease manifestations and improved macroscopic and microscopic inflammation. This preventive effect was found to be associated with increased release of anti-inflammatory cytokine IL-10 and decreased release of proinflammatory cytokines IFN-γ, TNF-α, IL-6 and IL-17 by the splenocytes of treated mice. From this study, it can be envisaged that pretreatment with filarial protein, rWbL2, can prevent the establishment of ulcerative colitis in BALB/c mice. The underlying immunological mechanism may involve the up-regulation of Th2 immune response with down-regulation of Th1 response.

Dermatoglyphics and karyotype analysis in primary amenorrhoea.

BACKGROUND Dermatoglyphics is the scientific study of the skin ridge patterns on the fingers, toes, palms of the hands and soles of feet. Dermatoglyphics is in use as a supportive diagnostic tool in genetic or chromosomal disorders as well as in clinical conditions with genetic etiologies. Primary amenorrhoea and Dermatoglyphics, both have the suspected multifactorial (genetic and environmental) aetiologies. OBJECTIVE In the present study the finger dermatoglyphic patterns were studied in primary amenorrhoea cases and age matched fertile control females and also attention was given to find out whether a specific dermatoglyphic trait exists in primary amenorrhoea cases and whether it was statistically significant. MATERIALS AND METHODS To study the role of dermatoglyphics in primary amenorrhoea, a study was conducted on 30 subjects with primary amenorrhoea (as cases) and compared it with equal number of age matched fertile females (as controls). We studied fingertip patterns in all the subjects enrolled. Simultaneously we have assessed the Karyotype of primary amenorrhoea cases. RESULT AND CONCLUSION Two subjects in experimental group have shown abnormal Karyotypes. The most significant finding in present study was increased total finger ridge count (TFRC) in primary amenorrhoea cases which was statistically significant. We also found higher frequency of loops and arches in primary amenorrhoea with abnormal karyotypes. This type of study may be quite useful as a supportive investigation, in stating the predisposition of an individual to primary amenorrhoea and referral of an individual for karyotyping.

Large Scale 7436-bp Deletions in Human Sperm Mitochondrial DNA with Spermatozoa Dysfunction and Male Infertility

INTRODUCTION Mitochondria and mitochondrial DNA are essential to sperm motility and fertility. It controls growth, development and differentiation through oxidation energy supply. Mitochondrial (mtDNA) deletions or mutation are frequently attributed to defects of sperm motility and finally these deletions lead to sperm dysfunction and causes infertility in male. AIM To investigate the correlation between large scale 7436-bp deletions in sperm mtDNA and non-motility of sperm in asthenozoospermia and Oligoasthenoteratozoospermia (OAT) infertile men. MATERIALS AND METHODS The present prospective study was carried out in Human Genetic Division, Department of Anatomy, Mahatma Gandhi Institute of Medical Sciences, Sevagram from June 2014 to July 2016. We have studied 110 asthenozoospermia and OAT infertile men whose semen profile indicated abnormal motility and 50 normal fertile controls. Of 110 infertile men, 70 had asthenozoospermia and 40 had OAT. Fractionations of spermatozoa were done in each semen sample on the basis of their motility by percoll gradients discontinuous technique. Long-range PCR was used for detection of 7436-bp deletions in sperm mtDNA and was confirmed by primer shift technique. RESULTS Overall eight subjects (8/110; 7.2%) of which six (6/70; 8.57%) asthenozoospermia and two (2/40; 5%) OAT had shown deletions of 7436-bp. In 40% percoll fraction had more non-motile spermatozoa than 80% percoll fraction. The non-motile spermatozoa in 40% percoll fractions showed more mtDNA deletions (7.2%) than the motile spermatozoa in 80% percoll fraction (2.7%). The sequencing of flanking regions of deleted mtDNA confirmed 7436-bp deletions. Interestingly, no deletions were found in control subjects. CONCLUSION Though, the frequency of 7436-bp deletions in sperm mtDNA was low in infertile cases but meaningful indications were there when results were compared with controls. It is indicated that large scale deletions 7436-bp of mtDNA is associated with abnormal sperm motility. The 7436-bp deletions of mtDNA in spermatozoa may be one of the important causes of dysfunction and non-motile sperm.

Human filarial proteins attenuate chronic colitis in an experimental mouse model

Encouraged by our earlier results of promising therapeutic effect of filarial recombinant proteins BmALT2, BmCys and WbL2 individually in the mouse model of acute ulcerative colitis, in this study, these proteins have been explored individually and in different combinations for their therapeutic potential in dextran sulphate sodium (DSS)‐induced chronic colitis mice. These mice, treated with filarial proteins, showed reduced disease parameters including body weight loss, disease activity index, macroscopic and histopathological scores of colon and myeloperoxidase activity in colonic mucosa. Among various treatment schemes, rBmALT2 + rBmCys which showed most pronounced therapeutic implication was found to downregulate the mRNA expressions of IFN‐γ and TNF‐α and upregulate IL‐10 and TGF‐β expression in the splenocytes. Also, increase in level of IgG1 and IgG2a isotypes in the sera of rBmALT2 + rBmCys‐treated colitis mice was noted. Activated NF‐κB level was found to be reduced in the colon of treated colitis mice compared to untreated one. In conclusion, filarial proteins in combination have been shown to improve the clinicopathologic status of chronic colitis through suppression of pro‐inflammatory immune response most possibly in NF‐κB‐dependent manner. We propose this therapeutic strategy to be tested further to be considered as an effective option in chronic colitis.

Immunomodulatory potential of recombinant filarial protein, rWbL2, and its therapeutic implication in experimental ulcerative colitis in mouse

Abstract Objective: Immunomodulation by helminth proteins has potential therapeutic implications in inflammatory bowel disease. In the present study, we have explored the therapeutic effect of a RAL family protein of filarial parasite Wuchereria bancrofti i.e., rWbL2 protein against DSS induced colitis in a mouse model. Materials and methods: Anti-inflammatory activity of rWbL2 on mice peritoneal exudate cells was analyzed under in vitro condition. The colitis mice were treated intraperitoneally (i.p.) with rWbL2 in increasing doses (10 µg, 25 µg, and 50 µg) on days 4, 5, and 6. Disease severity was assessed by disease activity index (DAI), macroscopic and histopathological scores, and enzyme myeloperoxidase activity (MPO) in the colon. The response of the cultured splenocytes from treated mice to Con-A stimulation, in terms of ELISA-based assessment of the protein followed by the assessment of mRNA expression of cytokines, was measured by real-time PCR analysis. Result: rWbL2 protein showed anti-inflammatory activity in vitro. Treatment with rWbL2 (at 25 µg/dose) effectively attenuated disease severity by reducing weight loss, DAI, mucosal edema, colon damage, and MPO activity. This therapeutic effect was found to be associated with increased release of anti-inflammatory cytokine IL-10 and decreased release of pro-inflammatory cytokine IFN-γ and TNF-α by the splenocytes of treated mice followed by stimulation with Con-A. Conclusions: These results provide evidence of the strong immunomodulatory potential of rWbL2 protein implicating its therapeutic application against ulcerative colitis.

Histomorphometric demonstration of the effect of chronic use of nonsteroidal anti-inflammatory drugs–ibuprofen on mucosa of small intestine

The purpose of this study was to ascertain change in structure of mucosa of small intestine, if any, in small intestine of Swiss albino mice as an effect of chronic use of nonsteroidal anti-inflammatory drugs–Ibuprofen. Longitudinal study conducted on 46 adult Swiss albino mice, 23 as experimental and 23 as control. Ibuprofen was given at a dose of 40 µg/g body weight per day for 6 weeks by intragastric route in experimental group of mice while control group of mice received same volume of distilled water. Mice of both the groups were sacrificed and desired segments of small intestines were dissected out and tissues were subjected to histological processing. Histomorphometry was performed and relevant photomicrographs were obtained. Students unpaired t test by GraphPad Prism 6 software. Height of villi was not significantly altered but there was significant reduction of the number of goblet and non-goblet cells (enterocytes and other columnar cells) in mucosal lining of the small intestine of experimental group of mice. Percent distribution of the goblet and non-goblet cells was not altered in villi of two groups. Chronic exposure of Ibuprofen in therapeutic dosage caused reduction of the functional cell mass in lining epithelium of villi of middle segment of small intestine. However, there was no evidence of ulcerative or hemorrhagic lesion.

Relation between Renal Length and Renal Volume with PatientâÂÂs BMI: A CriticalAppraisal

In urological and nephrological practices, evaluation of kidney size imparts a valuable diagnostic parameter. Age, gender, body mass index, pregnancy and co-morbid conditions such as diabetes mellitus, hypertension are supposed to affect the renal size. Measurements of renal dimensions can be carried out by using different modern techniques like ultrasonography, CT scan and MRI. On the other hand body mass index (BMI) provides information to know the patient’s obesity which is based on patient’s height and weight. Information available from one particular region may not satisfy the other region as renal parameters varies with different ethnic group and body size. In this prospective, reviewing literature of different studies evaluated so far to predict the relationship between renal dimensions and BMI is carried out. These studies revealed that most of renal parameters were positively correlated with body size of an individual, therefore they can be used to estimate the size and volume of kidney. Moreover, the standard normal reference ranges for renal volume and size can serve as gold standard and acts as an adjunct to judge the atrophic or hypertrophic condition of kidney. Hence, establishing relationship between measured renal dimensions and BMI will serve as a useful guideline for detection of diseased condition of kidney. This paper sets out to present published data on researches that has fortified knowledge and understanding of correlation between different renal parameters and BMI, keeping in mind different demographic background. Thus here we are putting a comprehensive account of the studies carried out globally to establish the inter-relationship between renal dimensions with BMI which definitely helps the nephrologists in early diagnosing the renal diseases.

Unilateral isolated incompletely duplicated ureter

The aim of this study was to report a congenital anomaly in a cadaveric dissection. During routine undergraduate dissection in a middle-aged male cadaver, we found that on the left side, there was a presence of an incompletely duplicated ureter. On the right side the ureter was single in its whole extent. No other congenital anomaly was found to be associated with this. The two limbs of the left ureter joined at about a distance of 5 cm from the bladder wall. A duplicated ureter is commonly found in association with other congenital anomalies and defects. The present case report describes a rare case of an isolated duplicated ureter with a normal kidney, urinary bladder, and renal vessels. This case report adds on to the literature and will be helpful and interesting for the surgeons. The possible embryological reasons for the formation of a duplicated ureter will be discussed.