Normal Genitalia Research Articles

Sex-mismatched after stem cell transplant: A case report

Introduction Ovarian failure associated with stem cell transplantation is generally awarded to total body irradiation and chemotherapeutic agents. The particularity of this observation is that describes a change in cell line in our patient following bone marrow transplant from her HLA matched brother. Case report A 15-year old girl was referred to our department of endocrinology with secondary amenorrhea. She was diagnosed with lymphoblastic Leukemia at the age of 13. She received chemotherapy and total body radiation after which she had bone marrow transplantation. Patient had menarche at 12 years and 9 months, she had regular menstruations for 5 months. Breast development was automatically since the age of 10 years and physical examination showed normal female genitalia, Tanner stage 3. The hormonal profile showed elevated FSH and LH, low estrogen, and normal prolactin, confirming the POI. Pelvic ultrasound showed a pubertal uterus and two small ovaries without follicles. Bone age was 14 years. Chromosomal analysis was 46, XY. The cytogenetic analysis carried out on a skin fibroblasts which showed a normal female karyotype identic with her chromosomal analysis before transplantation. Conclusion Ovarian failure is common after allo-stem cell transplant (SCT). It is usually secondary to POI due to chemotherapy. Sex-mismatched after SCT is defined when there is a male donor and female recipient and vice versa, it does not affect fertility. Only the karyotype of hemocyte is 46 XY, but other somatic cells and germ cells are 46 XX as somatic and germ cells originated during the embryonic stage.

640 Medico-legal evaluation of patients with genital disorders in Upper Egypt: a preliminary data

In Upper Egypt, the Forensic Medicine Authority of the Ministry of Justice transfers patients with genital disorders for proper Andrological evaluation, which helps in making its decisions according to the Law. Therefore, our study aims to evaluate the types of cases referred for andrological evaluation, their clinical, laboratory, imaging characteristics and finally the decisions of these cases. The study included 15 patients with genital disorders who referred from the forensic medicine authority for evaluation. Initial evaluation was done through history taking, general & genital examination. Hormonal profile including total Testosterone, FSH, LH, Prolactin, Estradiol (E2), and random blood sugar were done. Karyotyping, scrotal and/or penile color Doppler were done when appropriate. Data was presented as median (range), or percentage (%) when appropriate. Patients' characteristics showed age 29.5 (1-46) years. Most patients were married 73.3% (11/15). Cause of referral was mainly for erectile function evaluation due to marital conflicts 60%. (9/15). Other causes included sex determination 20% and genital assault 20%. All cases were free from medical co-morbidity (Diabetes, Hypertension) except one patient was with Dyslipidemia. General examination was normal in most patients 80% (12/15). Abnormal genital examination was noted in the form of hypospadias 13.3% (2/15), atrophic testis 26.6% (4/15), and varicocele 40% (6/15). Patients hormonal profile showed total testosterone 4.6 (3.69-7.82) ng/ml, FSH 3.53 (0.6-59.4) mIU/ml, LH 4.6(1.2-23.8) mIU/ml, prolactin 12.55 (4.9-18) ng/ml, and E2 43 (24-56.9) pg/ml. As regards the erectile function evaluation, patients with normal evaluation, venogenic, arteriogenic erectile dysfunction 26.7% (4/15), 26.7% (4/15), 6.6% (1/15) respectively. Other patients' diagnosis included normal genitalia, atrophic testis, and male pseudo hermaphrodite.

The Penile Perception Score after distal hypospadias repair with tubularized incised plate and Mathieu

Objective: The aim of this study was to evaluate the Penile Perception Score (PPS) after repair of distal penile hypospadias using tabularized incised plate (TIP) and Mathieu procedures. Methods: A randomized controlled study was conducted at urology department from October 2013 to May 2014. It included 60 children who were divided into three groups: group A included 20 patients with distal hypospadias corrected using TIP; group B included 20 patients with distal hypospadias corrected using Mathieu; and group C included 20 children with normal male genitalia who were circumcised and considered as a control group. Results: There were no statistically significant differences between group A and group B in urologists’ and parents’ average sum of PPS evaluation, but urologists’ satisfaction was more than parents’ satisfaction. The PPS for TIP was 8.43 and 7.80 for urologists and parents, respectively. The difference was significant. The PPS for Mathieu were 8.04 and 7.89 for urologists and parents, respectively, with no significance. There were no statistically significant differences between group A and group B in the postoperative evaluation by urologists and by parents in meatus, glans, shaft skin, and general appearance. There were complications in 10% of cases from group A in the form of fistula, in 30% of cases from group B in the form of fistula in 25% and meatal stenosis in 5%, and no complication in group C. Conclusion: TIP showed better PPS score compared with Mathieu in hypospadias repair. However, the results of this study were not statistically significant. PPS was a good scale to evaluate hypospadias repair after surgery. Keywords: hypospadias, parents and urology, Penile Perception Score

Familial Glucocorticoid Deficiency Presenting as Progressive Hyperpigmentation: A Case Report

Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive disease caused by resistant of ACTH receptor at adrenal cortex leading to (usually) isolated glucocorticoid deficiency with normal mineralocorticoid secrete. Patients with FGD usually presented in neonatal–childhood period with signs/symptoms of glucocorticoid deficiency such as hypoglycemia, hyperpigmentation, Failure to thrive, shock and death if treatment was delayed. Labs usually revealed high ACTH, low cortisol but normal 17 OHP, electrolyte, androgen. Here we describe a 3-yearsold Saudi girl, with history of progressive hyperpigmentation for first year of life, but no history of hypoglycaemia or neonatal jaundice, no history of a lacrimation or dysphagia and positive similar family history. She had generalized Hyperpigmentation with normal female genitalia. Her cortisol was low with high ACTH level, but normal electrolyte 17, Hydroxyprogesterone, aldosterone, renin, androgen. Familial Glucocorticoid Deficiency was diagnosed, and maintenance dose of hydro cortisol was started, and patient pigmentation was improved few week latter.

Clinical and molecular cytogenetic characterization of four unrelated patients carrying 2p14 microdeletions.

We report the clinical and molecular cytogenetic characterization of four unrelated patients from France and Spain, carrying 2p14 microdeletions and presenting with intellectual disability and dysmorphisms. 2p14 microdeletions are very rare. Seven patients have been reported so far harboring deletions including 2p14p15 and encompassing OTX1, whose haploinsufficiency is frequently associated with genitourinary defects. To date, only one patient has been reported carrying a more proximal 2p14 microdeletion which does not include OTX1. Here, we report three further patients carrying proximal 2p14 microdeletions not including OTX1 and one patient carrying a more distal 2p14p15 microdeletion including this gene, providing new insights into the associated phenotypic spectrum. First, our study and a review of the literature showed that 3/4 patients carrying proximal 2p14 microdeletions had sensorineural hearing loss, suggesting the presence of a previously unreported deafness-causing gene in this chromosomal region. Second, one patient developed a progressive cardiomyopathy, suggesting that a cardiac follow-up should be systematically warranted even in the absence of congenital heart disease. We speculate that ACTR2 and MEIS1 might respectively play a role in the pathogenesis of the observed deafness and cardiomyopathy. Third, we observed other previously unreported features such as glaucoma, retinopathy, and mild midline abnormalities including short corpus callosum, hypospadias and anteriorly placed anus. Finally, the patient carrying a 2p14p15 deletion including OTX1 had normal kidneys and genitalia, thus confirming that OTX1 haploinsufficiency is not invariably associated with genitourinary defects. In conclusion, our study contributes significantly to delineate the phenotypic spectrum of 2p14 microdeletions.

Restoring normal anatomy in female patients with atypical genitalia

Female patients with congenital adrenal hyperplasia (CAH) have varying degrees of atypical genitalia secondary to prenatal and postnatal androgen exposure. Surgical treatment is focused on restoring normal genitalia anatomy by bringing the vagina to the normal position on the perineum, separating the distal vagina from the urethra, forming a normal introitus and preserving sexual function of the clitoris by accepting moderate degrees of hypertrophy as normal and strategically reducing clitoral size only in the most severely virilized patients. There remains a need for continued monitoring of patients as they go through puberty with the possibility of additional surgery for vaginal stenosis. Anatomically based surgery and refinement in surgical techniques with acceptance of moderate degrees of clitoral hypertrophy as normal should improve long-term outcomes.

Campomelic dysplasia with dextrocardia and without sex-reversal

Campomelic dysplasia (CD) is a very rare, sporadic, autosomal dominant syndrome. CD is characterized by an association of skeletal (bowed long bones, pelvis and chest abnormalities, eleven rib pairs) and extraskeletal abnormalities (facial dysmorphism, sex-reversal in boy and visceral malformations). We report a case of a 3 days old male diagnosed by CD. At birth, the boy displayed severe asphyxia and therefore needed intensive care. Physical examination revealed short stature, a craniofacial dysmorphism, cleft palate, muscle hypotonia, skeletal anomalies (chest hypoplasia, curved short upper and lower limbs, talipes equinovarus ) cardiac malformations, normal male genitalia. The X-ray examination showed bell-shaped, narrow thoracic cage, hypoplastic scapulae, reduced cranial, pelvic, tibial and fibular ossification, absence of nasal and sternal ossification, cervical kyphosis, 11 pairs of ribs, flat and short vertebral bodies, short humerus with widened distal epiphysis, anterolateral femoral and tibial bowing and ankle valgus deformity. Dextrocardia was identified on X-ray examination and on echocardiography. Karyotype was 46,XY. The particularities of cases are campomelic dysplasia with dextrocardia, but without sex-reversal.

Shorter anogenital distance correlates with the severity of hypospadias in pre-pubertal boys.

Do pre-pubertal boys with hypospadias have a shorter anogenital distance (AGD) than boys with normal genitalia? AGD is significantly shorter in boys with hypospadias and decreases with the severity of hypospadias. Animal studies have shown that androgen disruption and exposure to endocrine disrupting chemicals during a critical time period in early gestation, termed the male programming window (MPW), result in hypospadias and reduced AGD; and the severity of hypospadias correlates with the reduction in AGD. However, this correlation has not been established in humans. A prospective descriptive study involving measurement of AGD in pre-pubertal boys (n = 458) presenting to our pediatric urology clinic with hypospadias and normal genitalia was performed over a period of 3 years. AGD was measured in pre-pubertal boys from 5 months to 14 years of age presenting to our clinic with hypospadias (n = 180: four were excluded) and compared with randomly selected boys with normal genitalia (controls, n = 274). Three variants of AGD, from the midpoint of the anus to base of the scrotum (AGD-AS), to the anterior base of penis (AGD-1) and to the posterior base of penis (AGD-2), were measured and assessed for correlation with the severity of hypospadias. Severity of hypospadias was classified as anterior, middle and posterior according to the meatal location. No significant difference in weight (P = 0.123), age (P = 0.162) or height (P = 0.591) between the two groups was observed. Only AGD-AS was significantly shorter in boys with hypospadias compared with controls (mean ± SD: 40.6 ± 9.7 mm versus 45.6 ± 9.4 mm, P < 0.001). This relation persisted after adjusting AGD for weight, height and age (β = 0.016, 95% confidence interval: 0.10-0.21; P < 0.001). The Spearman test showed a significant negative correlation for the severity of hypospadias with all the three AGD measures. Analysis of variance between anterior, middle and posterior subgroups showed a significant reduction in mean AGD-AS (P = 0.003) and AGD-2 (P = 0.008). No data were collected pertaining to in utero exposure to endocrine disrupting chemicals (EDCs) or cigarette smoke, or current diet and environmental exposure to EDCs, which may have influenced the AGD. Family history of genital malformation and use of IVF were not known. There may have been a selection bias as only boys presenting to our clinic were included. The findings suggest that prenatal androgens during early gestation influence development of the male reproductive system and support the existence of a MPW in humans. Of the three AGDs, AGD-AS may be the most reliable biomarker of this in utero androgen action. However, no direct link to any specific exposure leading to shortened AGD in pre-pubertal boys with hypospadias could be determined. Further large scale multi-center studies are needed to understand this association better. Funding was from the Hypospadias Foundation. No conflicts of interest to disclose.

Gonadal and Epididymal Sperm Reserves of Yankasa Rams Treated with Cypermethrin

Cypermethrin 3% (30 gram/liter) at the dose rate of 3mg/kg (0.1ml/kg) was used as pour-on. Sixteen healthy Yankasa rams with clinically normal genitalia were used. The animals were divided into groups A and B i.e. experimental and control groups. The experimental group was given Cypermethrin treatment as pour-on fortnightly for a period of twelve weeks. The control group was given distilled water at the same rate (0.1ml/kg) and route of administration for the same length of time. Rams from both the experimental and control groups were sacrificed for estimation of gonadal and epididymal sperm/spermatids reserves at the end of the twelve week studies. Epididymal and gonadal sperm reserves were estimated using a haemocytometer. Results showed that gonadal sperm reserves was not significantly different between the two groups (P>0.05). Epididymal sperm reserves showed that there was a significant difference between the cauda epididymis of the experimental and the control rams with the experimental rams showing lower cauda epididymal reserves of 149.75±13.13 x106/g while the control rams had a value of 244.56±17.09 x 106/g (P<0.05), the caput and corpus epididymis were not significantly different. It was concluded that 3% Cypermethrin at the dose rate of 3mg/kg given for a period of twelve weeks affects the epididymal sperm reserves at the cauda segment. It was therefore recommended that the use of Cypermethrin be applied with caution in rams as it may affect the fertility of Yankasa rams negatively.

Thyroid Pathology of Cypermethrin and Its Reproductive Implications in Yankasa Rams

An investigation into the pathological effects of Cypermethrin on the thyroid gland of Yankasa rams was carried out. Sixteen sexually-mature, healthy Yankasa rams aged 18 - 30 months and weighing between 21.5 - 46.5kg with clinically normal genitalia were used. They were divided equally into two groups (A and B). A served as the treatment group while B served as the control. (A) were given Cypermethrin (3%) at the dose rate of 3mg/kg (0.1ml/kg) body weight, topically as pour-on. (B)were given distilled water at the same dose rate of 0.1ml/kg body weight topically as pour-on. These treatments were repeated every two weeks for a period of 12 weeks. The rams were sacrificed at the end of 12 weeks and the following organs (liver, heart, kidney, brain and thyroid gland), were collected for gross and histopathology. Results showed that no gross pathologic lesions were found on the thyroid gland, theliver, brain and the kidneys of both groups. The heart of the treated group showed petechial hemorrhage. The mean weights of the following organs; liver, thyroid gland, kidneys and heart of the treated and the control groups were 400.00±0.00g and 420.00±20.0g; 2.30±0.37g and 1.50±0.31g; 70.20±9.90g and 72.60±3.70;127.51±46 and 100.00±20.00respectively. The differences in weight between the two groups were not significantly different (P>0.05). Histologic sections of the thyroid glands of the treated group showed hyperplasia of the follicular epithelial cells followed by collapsing of the follicles. Histologic sections of the cardiac muscles of the treated group showed focal infiltration of lymphocytes and loss of striations (degeneration). The liver, the kidneysand thebrain (cerebrum) of both groups showed no histologic lesions. It was concluded that Cypermethrin induced thyroid and cardiac pathology in the rams. It was recommended that more investigation be done to unravel the mechanism of inducing thyroid and cardiac pathology in rams.

Human MAMLD1 Gene Variations Seem Not Sufficient to Explain a 46,XY DSD Phenotype.

MAMLD1 is thought to cause disordered sex development in 46,XY patients. But its role is controversial because some MAMLD1 variants are also detected in normal individuals, several MAMLD1 mutations have wild-type activity in functional tests, and the male Mamld1-knockout mouse has normal genitalia and reproduction. Our aim was to search for MAMLD1 variations in 108 46,XY patients with disordered sex development, and to test them functionally. We detected MAMDL1 variations and compared SNP frequencies in controls and patients. We tested MAMLD1 transcriptional activity on promoters involved in sex development and assessed the effect of MAMLD1 on androgen production. MAMLD1 expression in normal steroid-producing tissues and mutant MAMLD1 protein expression were also assessed. Nine MAMLD1 mutations (7 novel) were characterized. In vitro, most MAMLD1 variants acted similarly to wild type. Only the L210X mutation showed loss of function in all tests. We detected no effect of wild-type or MAMLD1 variants on CYP17A1 enzyme activity in our cell experiments, and Western blots revealed no significant differences for MAMLD1 protein expression. MAMLD1 was expressed in human adult testes and adrenals. In conclusion, our data support the notion that MAMLD1 sequence variations may not suffice to explain the phenotype in carriers and that MAMLD1 may also have a role in adult life.

FLUSHING, SUCKLING AND SEXUAL MATURITY OF SMALL RUMINANTS FED DIET CONTAINING SILAGE OF TRITICALE, BERSEEM OR ITS MIXTURE

objective of this study was to investigate the effect of three types of silage (triticale, berseem and their combination) supplemented to concentrate fed mixture (CFM) on productive, as suckling milk amount, growth rate of newborns and reproductive efficiency, as fertility rate and sexual development of male lambs to be used as sires. Two small ruminate breeds (ewes and does) were assigned into three treatment groups. Forty-two ewes (n=14 ewe in each treatment) and fifty-seven does (n=19 does in each treatment). All animals were received diets at 21 days pre-mating season (as flushing diet) up to weaning. The feeding treatments (T) offered as following: T1 contained 60% CFM + 40% triticale silage, T2 included 60% CFM + 40% admixture silage (50% triticale and 50% berseem) and T3 contained 60% CFM + 40% berseem silage. To determine suckling milk amount of maternal; twelve ewes (n=4 in each treatment) and eighteen does (n=6 in each treatment) symmetrical in reproductive and productive characteristics were used. The maternal in suckling treatment were received the same T1, T2 and T3 diets mentioned above. In respect of the newborn growth rate, all suckling lambs and kids from birth up to weaning were used. In sexual maturation evaluation, fifteen healthy Rahmani lambs with normal genitalia, aged 150 days and at initial body weigh (22.21±0.42 kg) were allocated into three treatments; five lambs / treatment. Results showed that ewes and does in T2 group had a higher reproductive performance, parity patterns and reproductive ability than those in other treatments (T1 and T3). The clearly differences wereobserved among the dietary treatments during suckling period of ewes and does. The maternal fed on T2 produced more (P<0.05) amounts in suckling milk than those maternal fed T1 and T3. Consequently, T2 improved (P<0.05) newborns growth rate compared to newborns in either T1 or T3 groups. Furthermore, T2 group achieved better (P<0.05) sexual developments (i.e., age at puberty, body weight, scrotal circumference, testicular diameter, testicular length, testicular volume and semen characteristics at puberty) than T1 and T3 groups. These findings indicate that the inclusion of admixture silage has no adverse effects on productive and reproductive performance, suckling milk production and growth rate of newborns. Admixture silage causes earlier sexual maturation and goodness semen parameters of Rahmani male lambs. Thus, mixing more than types of silage may be a promising strategy nutrition in the future.

Equine Genital Squamous Cell Carcinoma: In Situ Hybridization Identifies a Distinct Subset Containing Equus caballus Papillomavirus 2.

Equus caballus papillomavirus 2 (EcPV2) has been proposed as an etiologic agent for genital squamous cell carcinoma (SCC), the most common malignant tumor of the horse penis. EcPV2 is commonly detected by polymerase chain reaction (PCR) on normal horse genitalia; therefore, unraveling the virus' role in oncogenic transformation requires other methods of detection. In this study, a highly sensitive multiple-probe chromogenic in situ hybridization (ISH) technique was designed to recognize the E6/E7 oncogenes of EcPV2. ISH demonstrated abundant virus within 6 of 13 penile and preputial SCCs, whereas evidence of solar damage was found in 6 cases that were negative for EcPV2 by ISH. The ISH technique is valuable for studies of pathogenesis, since it demonstrates for the first time that the vast majority of neoplastic cells contain virus. Moreover, hybridization was present in all metastases examined, implying stability of E6/E7 expression in these clonal populations of neoplastic cells. This study contributes to the accumulating evidence for a causal role of EcPV2 in a subset of genital SCCs in horses.

Re-do surgery of the patients with feminizing genitoplasty in congenital adrenal hyperplasia

A successful feminizing genitoplasty depends on creating morphologically and functionally near normal genitalia. But it’s still a challenging problem in the pediatric age group to perform a fully satisfactory genitalia. Our aim is to evaluate necessity for redo genitoplasty procedures in patients with congenital adrenal hyperplasia that previously performed genitoplasty.

A report from #BlueJC: Does exposure to images of modified genitalia change our perceptions of normal genitalia?

A report from #BlueJC: Does exposure to images of modified genitalia change our perceptions of normal genitalia?

Effect of strategic nutrient supplementation on the reproductive performance of anoestrus crossbred cattle in Malwa region of Madhya Pradesh

In Malwa region of Madhya Pradesh anoestrus is a major problem in crossbred cattle under field conditions due to under feeding and non availability of balanced ration. Thirty anoestrus crossbred cattle {20 heifers (average body wt. 224.75 ± 7.41 Kg) + 10 cows (average body wt. 316.50 ± 15.48 Kg, milk yield 4.9 ± 1.25 litre/h/d)} having normal genitalia were selected randomly from ten villages of Indore district. Average daily feed intake of each animal was recorded and proximate principle, major (Ca and P) and trace elements (Fe, Zn, Mn, Cu and Co) in available feedstuffs were determined to find out nutrient availability. Deficiency of nutrients was calculated by comparing the standard requirements of the animals. A strategic nutrient supplement [soya-De oiled cake, sodium dihydrogen orthophosphate dihydrate, zinc oxide, copper sulphate, vitamin A & E] containing deficient nutrients was formulated on the basis of nutrient deficiency observed in rations of anoestrus cows. Supplementation study was carried out in the same thirty affected anoestrus crossbred cattle by dividing them into two groups of 15 (10 heifers + 5 cows) in each. One of them served as control (un-supplemented), while another was supplemented with strategic nutrient supplement (Treatment). This supplementation study was continued for a period of two months. During which individual animal was regularly examined for onset of estrus and artificially inseminated and after two months of the artificial insemination, pregnancy diagnosis of each animal was carried out per rectally to find out the conception status. Onset of estrus (control vs treatment group) in heifers (10% vs 70%) and lactating cows (20% vs 60%) and conception rate (control vs treatment group) in heifers (10% vs 50%) and lactating cows (20% vs 60%) were higher in the animals of supplemented group, while an additional advantage in the form of improvement (P< 0.05) in the body weight of heifers of treated group was also observed. Thus it can be concluded that strategic nutrient supplementation not only improved the reproductive performance of the crossbred anestrous heifers and cows, but it also improved the growth performance of the heifers.

What's normal? Influencing women's perceptions of normal genitalia: an experiment involving exposure to modified and nonmodified images

Examine women's perceptions of what is 'normal' and 'desirable' in female genital appearance. Experiment with random allocation across three conditions. Community. A total of 97 women aged 18-30years. Women were randomly assigned to view a series of images of (1) surgically modified vulvas or (2) nonmodified vulvas, or (3) no images. They then viewed and rated ten target images of surgically modified vulvas and ten of unmodified vulvas. Women used a four-point Likert scale ('strongly agree' to 'strongly disagree'), to rate each target image for 'looks normal' and 'represents society's ideal'. For each woman, we created two summary scores that represented the extent to which she rated the unmodified vulvas as more 'normal' and more 'society's ideal' than the modified vulvas. For ratings of 'normality,' there was a significant effect for condition (F2,94 =2.75 P=0.007, radj2=0.082): women who had first viewed the modified images rated the modified target vulvas as more normal than the nonmodified vulvas, significantly different from the control group, who rated them as less normal. For ratings of 'society's ideal', there was again a significant effect for condition (F2,92 =7.72, P<0.001, radj2=0.125); all three groups rated modified target vulvas as more like society's ideal than the nonmodified target vulvas, with the effect significantly strongest for the women who had viewed the modified images. Exposure to images of modified vulvas may change women's perceptions of what is normal and desirable. This may explain why some healthy women seek labiaplasty.

Upregulation of TLR-4 and proinflammatory cytokine transcripts as diagnostic indicator of endometritis in buffaloes

The aim of present study was to examine the relative mRNA transcription of toll-like receptor 4 (TLR-4), CD14, interleukin-1β (IL-1β), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-α) in endometritic buffaloes. Reproductive tracts were collected from 60 buffaloes at postmortem. The genitalia were diagnosed for endometritis based on positive white side test of uterine fluid, presence of >5% polymorphonuclear cells in uterine cytology, and endometrial histopathology. The endometritis-positive genitalia were categorized as luteal (n = 10) and follicular (n = 10) based on the presence of corpus luteum. Further, 20 normal genitalia (luteal, n = 10 and follicular, n = 10) were also included as control. The endometrial tissue scrapings were collected ex vivo from all the samples, total RNA was extracted, complementary DNA was transcribed for each sample, and relative quantification of mRNA was done by real-time polymerase chain reaction. The threshold cycle values of all the samples were normalized with the housekeeping β-actin gene and relative fold change was calculated using 2−ΔΔCt method. Present study revealed a variable upregulation of TLR-4, CD14, IL-1β, IL-8, and TNF-α transcription in endometritic buffaloes either at luteal or follicular stage. Predominantly at luteal stage, the significant upregulation of TLR-4, IL-8, and TNF-α transcripts was 4.66-, 21.19-, and 42.49-fold, respectively, indicated the possibility of cytokine-based diagnostic method for the early indication of endometritis before the onset of cyclic estrus. Nevertheless, further research in respect to threshold level of each cytokine in uterine biopsies involving more number of animals is required to establish a potential marker gene(s) for the diagnosis of endometritis and for devising novel therapeutic intervention.

Y chromosome in Turner syndrome: detection of hidden mosaicism and the report of a rare X;Y translocation case

Turner syndrome (TS) is a common genetic disorder in females associated with the absence of complete or parts of a second sex chromosome. In 5-12% of patients, mosaicism for a cell line with a normal or structurally abnormal Y chromosome is identified. The presence of Y-chromosome material is of medical importance because it results in an increased risk of developing gonadal tumours and virilisation. Molecular study and fluorescence in situ hybridisation approaches were used to study 74 Brazilian TS patients in order to determine the frequency of hidden Y-chromosome mosaicism, and to infer the potential risk of developing malignancies. Additionally, we describe one TS girl with a very uncommon karyotype 46,X,der(X)t(X;Y)(p22.3?2;q11.23) comprising a partial monosomy of Xp22.3?2 together with a partial monosomy of Yq11.23. The presence of cryptic Y-chromosome-specific sequences was detected in 2.7% of the cases. All patients with Y-chromosome-positive sequences showed normal female genitalia with no signs of virilisation. Indeed, the clinical data from Y-chromosome-positive patients was very similar to those with Y-negative results. Therefore, we recommend that the search for hidden Y-chromosome mosaicism should be carried out in all TS cases and not be limited to virilised patients or carriers of a specific karyotype.

Results from 28 years of newborn screening for congenital adrenal hyperplasia in Sapporo City

Newborn mass screening (MS) for congenital adrenal hyperplasia (CAH) was started in 1989 at nationwide of Japan. To date, the goals for prevention of life-threatening salt-wasting crises and prevention of incorrect sex assignment in virilized females are almost achieved. However in Sapporo city, which is located in the northern part of Japan, a pilot study was started in 1982 after the assessment of its feasibility and efficiency. Here we report the results of MS from 1982 to 2010 in Sapporo city. In 28 years, 17-hydroxyprogesterone (17-OHP) was determined in MS samples in 498,147 newborns and a total of 2,466 screened newborns had abnormal 17-OHP. Among them, 26 patients were diagnosed with 21-hydroxylase deficiency (21-OHD), which corresponds to a prevalence of 1:19,160 live birth in Sapporo city. It is almost identical to the worldwide incidence of CAH. Among 26 patients, 20 patients were classified into salt-wasting forms, 5 patients were classified into simple virilizing forms and one was classified into nonclassic forms. 19 patients were female and 7 patients were male. Two familial cases were detected. The mean examination day of 25 patients (data of one patient was not available) was 6.4 days after birth (from 0 to 18 days). Among 18 female patients, 11 patients were clinically suspected because of virilization prior to MS result becoming available and were referred to neonatal center. 4 female patients were referred after MS screening. One female patient was assigned to male after birth; however the patient was diagnosed as having 21-OHD by MS and the sex was changed after the diagnosis. One female patient was a familial case and during pregnancy prenatal treatment was done and her genitalia was normal female. Another one patient having normal female genitalia and no symptom, was diagnosed as nonclassic forms by MS. By contrast to female patients, all 7 male patients were detected by MS. Another type of CAH was not detected in our study. In conclusion, MS for CAH in Sapporo city can be considered reliable.