Mutational spectrum of the TYR and SLC45A2 genes in Pakistani families with oculocutaneous albinism, and potential founder effect of missense substitution (p.Arg77Gln) of tyrosinase.

BACKGROUND: Oculocutaneous albinism (OCA) is an autosomal recessive disorder of abnormal melanin formation, which results in hypopigmentation of skin, hair and eyes. OCA is classified into four types based on clinical and genetic findings. OCA1 is the most severe form of albinism, and is caused by mutations in the tyrosinase (TYR) gene, while OCA4 is caused due to mutations in SLC45A2. METHODS: In total, 13 families with ≥ 3 members with OCA were enrolled. Family history was ascertained and pedigrees were drawn up. Blood samples were collected and processed for DNA extraction. Linkage analysis was performed by typing three short tandem repeat markers in candidate regions of TYR and SLC45A2. Sequence analysis was performed of all the coding exons and adjacent intronic sequences of both genes. RESULTS: Eight families showed linkage to OCA1 and one family showed linkage to OCA4. Four missense substitutions (p.Arg239Trp, p.Ser192Tyr, p.Ser44Arg and p.Arg77Gln) were identified in TYR in the families with OCA1 linkage, and another missense substitution (p.Gln272Lys) was identified in the family with OCA4 linkage. One of the identified missense substitution (p.Arg77Gln) in TYR was found in five different families, which had a common haplotype. CONCLUSIONS: We identified four missense substitutions in TYR and a single missense substitution in SLC45A2. One missense substitution (p.Arg77Gln) in TYR was found in five different families that originated from the same geographical area and displayed a common haplotype, suggesting a single origin that then spread to different geographical areas of Azad Kashmir, Pakistan.

A highly sensitive poly(chrysoidine G)-gold nanoparticle composite based nitrite sensor for food safety applications.

This work demonstrated the development of conducting poly(chrysoidine G) (PCG)-gold nanoparticle (AuNP)-modified fluorine-doped tin oxide (F : SnO2, FTO) film-coated glass electrodes for the sensitive electrochemical detection of nitrite (NO2-). The homogeneously distributed PCG nanoparticle layer was deposited onto the FTO electrode by cyclic voltammetry sweeping. AuNPs were then anchored onto the PCG/FTO electrode by the chemical reduction of pre-adsorbed Au3+ ions. The as-prepared AuNP/PCG/FTO electrode exhibited excellent electrocatalytic activity for the oxidation of NO2- with high sensitivity (approximately 0.63 µA cm-2µM-1) and a low limit of detection (0.095 µM), which is relevant within the normal concentration range of NO2- in human bodily fluids. The AuNP/PCG/FTO sensor showed sufficient reproducibility, repeatability, low interference, and strong recovery for NO2- detection in food samples. These results indicate that the AuNP/PCG nanocomposites have immense potential for the electrochemical detection of other biologically important compounds.

ASSESSMENT OF RADON IN SOIL AND WATER IN DIFFERENT REGIONS OF KOLHAPUR DISTRICT, MAHARASHTRA, INDIA.

Researchers have already established that inhalation of high radon concentration is hazardous to human health. Radon concentration has been measured in water and soil, in various part of Kolhapur district has been carried out by the AQTEK Smart RnDuo which is an active device technique. The observed minimum value of the radon mass exhalation rate of the soil is 13.16 ± 0.83 mBq/kg/h and maximum is 35.11 ± 1.84 mBq/kg/h. The minimum value of the Radon concentration in water is 0.33 ± 0.052 Bq/L and maximum is 7.32 ± 0.078 Bq/L. These values of radon concentration are below the action of recommended level by the USEPA, which is set as the maximum contaminant level of 11.1-148 Bq/L of radon in drinking water. Total annual effective dose rate of water is 11 µSv/y. The purpose of present study is to assess radiological risk from consumption of water that provide in Kolhapur district and to evaluate the radon mass exhalation rate of soil in few places of Kolhapur district.

Rationale and design for the CARDIOVIT Study (Cardiovit, Atherosclerotic vascular disease and hypeRhomocysteinemia: an epiDemiological study in Indians, additionally evaluating the effect of Oral VITamin supplementation).

BACKGROUND AND RATIONALE: Atherosclerotic vascular disease (AVD), including coronary heart disease (CHD), is on the rise in India. Mortality due to CHD is estimated to be 40% higher in Indian Asians compared with Europeans. Hyperhomocyst(e)inaemia (hypertHcy) is emerging as a novel independent risk factor for AVD. Moreover, there is increasing evidence of its association with CHD in subjects of Indian origin possibly because of inadequate folic acid and B(12) status. However, there is a lack of epidemiological data on its prevalence in Indians. A need to evaluate the optimum dose of folic acid required to reduce plasma homocyst(e)ine (tHcy) also exists. OBJECTIVES AND METHODS: The CARDIOVIT study, divided into two stages, is designed to determine the prevalence and relevance of hypertHcy in Indian subjects and also evaluate the tHcy-lowering effect of a marketed multivitamin formulation (Cardiovit). Stage I is a cross-sectional epidemiological study to estimate the prevalence of hypertHcy in a randomly selected population in South India. We planned to screen 2000 subjects between 25 and 64 years of age for established risk factors; plasma tHcy levels will be estimated and the correlation of AVD with established risk factors, plasma tHcy and nutritional factors was evaluated. Subjects having tHcy levels > 12 micromol/L are recruited in Stage II; a randomized, three-arm, double-blind, double-dummy, placebo-controlled, parallel-group, comparative study designed to evaluate the tHcy-lowering effect of a multivitamin formulation containing 300 microg folic acid, 1 microg vitamin B(12) and 1.5 mg vitamin B(6), along with Vitamins A, C, E and selenium. Subjects will be randomized into three groups (n = 64 completed subjects each) and receive either two placebos, or one placebo and multivitamin, or two multivitamin tablets for 6 weeks. Serum vitamin B(12), serum and red cell folate are estimated at baseline and at 6 weeks. Descriptive, univariate analysis, multiple logistic regression model and appropriate statistical tests will be used to analyse the prevalence of hypertHcy, association with risk factors of AVD, efficacy and safety of the multivitamin formulation and other data per protocol. CONCLUSIONS: CARDIOVIT is one of the first and largest studies in India to provide insight into the prevalence of hypertHcy, its association with AVD and conventional risk factors and the role of multivitamins in reducing it.

Experience with Extra Hepatic Intra Abdominal Hydatid Cyst.

INTRODUCTION: Hydatid disease is a significant health problem with their location at Extrahepatic Intra abdominal sites is a very rare disease. Disease in these sites usually found concurrently with liver hydatid disease. Diagnosis is based blood tests & imaging studies. Surgery is the mainstay of treatment. This study will help us in determining the frequency of Extra Hepatic Intra Abdominal Hydatid Disease. Also different methods to diagnose & treat the disease will be evaluated. METHODS: It includes patients of extra hepatic intra abdominal Hydatid cyst above 13 years and underwent surgery from 2009 to 2014. Variables include age, sex, clinical findings, diagnostic investigations, operative findings, operative procedure, post operative complications. RESULTS: Eleven patients; 07 male & 04 female; mean age 37.54 years. Most common symptoms were mass in abdomen in 11 patients & dull pain abdomen in 09 (81.8%) patients. Examination revealed non tender mass in epigastric & umbilical region in 04 (36.3%) patients. Ultrasound Abdomen showed hydatid cyst spleen in 02 (18.1%) patients, epigastrium in 04 (36.3%) patients, beneath left crus of diaphragm in 02 (18.1%) patients & right iliac fossa & pelvis in 1 (9.09%) patient. In 02 (18.1%) patients multiple Hydatid cysts were noted. Hydatid cysts liver found in 07 (63.6%) patients. C.T scan Abdomen was performed in 09 (81.8%) patients. Surgical procedures performed include Saucerization & omental packing in liver Hydatid cysts; Splenectomy for Splenic disease & complete excision of remaining intra-abdominal Hydatid cysts. Postoperative complications noted in 05 (45.4%) patients. CONCLUSION: Extra hepatic intra abdominal is an infrequent disease presents significant diagnostic & therapeutic challenge for surgeons.

Adrenal and ovarian steroid secretion in the rat estrous cycle temporally related to gonadotropins and steroid levels found in peripheral plasma.

This study was undertaken to determine the patterns of estrogen and progestin secretion by the adrenal and the ovary and to relate these to the levels of steroids and gonadotropins found in peripheral plasma. A total of 70 rats were used in this study. Adrenal venous (ADVB) and ovarian venous blood (OVB) were collected by concurrent cannulation and peripheral blood (PB) was obtained from the abdominal aorta. Estrone (E1), estradiol (E2), PROGESTERONE (P) and 17alpha-hydroxyprogesterone (17alpha-OHP) were measured by radioimmunoassay. The concentration of E2 in general was higher in ADVB than in OVB. Although increased levels of E2 in ADVB were found on the afternoon of proestrus (1527-2634 pg/ml), they were not significantly different from those recorded at other times. In contrast E2 in OVB was lowest at 1000-1200 hr of estrus (225 pg/ml) and highest at 1000-2000 hr of proestrus (1190-1605 pg/ml). The E2 in PB followed the pattern seen in OVB; the values ranged from 80-211 pg/ml. P concentration in general was higher in ADVB than in OVB. The lowest concentration of P in ADVB was between 0200-0400 hr of estrus (183 ng/ml) and the highest between 1400-1600 hr of proestrus (580 ng/ml) and 1000-1200 hr of estrus (591 ng/ml). The lowest concentration of P in the OVB was between 1600-1800 hr of diestrus (42 ng/ml) and the highest between 0200-0400 hr of estrus (466 ng/ml). In the PB, P reflected changes in adrenal as well as ovarian secretion; the values ranged fron 7-25 ng/ml. E1 in ADVB was in general lower than in OVB, and there were no significant changes at any stage of the cycle. In the OVB, however, E1 showed a pattern similar to that of E2. Levels of E1 in PB were very low and failed to show any significant changes. 17alpha-OHP was high in ADVB, although there were no significant changes during the cycle; PB levels also did not show any significant changes. In the OVB a peak of 17alpha-OHP was recorded between 0200-0400 hr on the day of estrus. The LH and FSH levels showed increases after 1600 hr on the day of proestrus and this was in agreement with earlier studies. Since the adrenal secretes all of the 4 steroids studied, it is reasonable to assume that it does exert some effect in the pituitary-gonadal axis. A role of adrenal P for the facilitation of the preovulatory LH release has been indicated and the increased secretion of adrenal P between 1400-1600 hr of proestrus supports this possibility.

The effect of an aromatase inhibitor, 4-hydroxy-4-androstene-3,17-dione, on estrogen-dependent processes in reproduction and breast cancer.

4-Hydroxy-f-androstene-3,17-dione (4-OH-A) when tested at various concentrations was found to inhibit markedly the conversion of 4-andorstene-3,17-dione to estrogens inhuman placental and rat ovarian microsomes. To obtain evidence that estrogen biosynthesis could also be reduced in vivo with 4-OH-A, rats were treated sc at a dose level of 50 mg/kg body weight. After 3 h the ovarian veins were cannulated and blood collected. Estradiol concentrations in the plasma were reduced by 80% compared to control values during the proestrous surge and on Day 4 of pregnancy. 4-OH-A was also found to be effective in controlling estrogen-dependent reproductive and neoplastic processes. In rats treated from Day 2-7 of pregnancy, implantation of fertilized ova was completely prevented in some rats, while in others either implantation was delayed or the development of implants was retarded. 4-OH-A treatment of rats having estrogen-dependent breast tumors induced by 7,12-dimethylbenz(a)anthracene caused 80% of the tumors to regress significantly in 4 weeks of treatment; 42% of these regressed completely.

Synergism of estrogen and bromergocryptine in the induction of luteolysis in cynomolgus monkeys (Macaca fascicularis).

Sequential administration of estrogen (40 microgram depoestradiol cypionate and estradiol benzoate) on cycle days 18--20 and bromergocryptine (CB-154; 0.5 mg/kg, twice daily) on cycle days 20-22 induced luteolysis in seven of nine cynomolgus monkeys, significantly shortened menstrual cycles (23.5 +/- 0.7 days), and caused a significant decline in plasma progesterone levels during the period of CB-154 administration. Two treated monkeys did not have shortened menstrual cycles, although a luteolytic effect on plasma progesterone was evident. One monkey with a late ovulation (confirmed by endocrine parameters and laparoscopic observation) was treated with the same regimen in the early luteal phase; this also resulted in a decline in plasma progesterone and a shortened luteal phase (11 days). Treatment with the estrogen or the CB-154 component alone had no effect on menstrual cycle length or plasma progesterone levels. Similarly, treatment with CB-154 throughout the luteal phase had no effect on luteal function. An abbreviated treatment consisting of 1 day of estrogen and 1 day of CB-154 administration gave some indication of a luteolytic effect. None of these treatments produced detectable side effects. These studies have demonstrated a synergism of estrogen and CB-154 in the induction of luteolysis in cynomolgus monkeys. The present regimen, because of the effectiveness and apparent lack of side effects, deserves further investigation for possible clinical application.

Atypical cribriform lesions on prostate biopsy.

Atypical cribriform lesions on prostate needle biopsy specimens are rare and difficult to diagnose. Of 574 high-grade prostatic intraepithelial neoplasia (PIN) lesions on needle biopsy seen at our institution over 75 months, we identified 23 consult cases in which the differential diagnosis was cribriform high-grade PIN versus infiltrating cribriform carcinoma. Prebiopsy prostate-specific antigen (PSA) averaged 6.5 ng/mL (range, 0.3 to 37.3). A positive digital rectal examination (DRE) was present in 12 of 22 (55%) patients for whom information was available. Ordinary high-grade PIN was present elsewhere in the biopsy specimens in 32% of cases. The following architectural features of cribriform glands were evaluated: number (mean, 5; range, 1 to 21); largest size (mean, 0.5 mm; range, 0.1 to 1 mm); necrosis (14%); detached cribriform fragments (18%); stromal fibrosis (18%); and bilaterality (22%). Cytologically, there was cellular maturation toward the center of the cribriform glands (45%); identifiable basal cells on hematoxylin and eosin sections (36%); marked nuclear atypia (9%); and mitoses (23%). Nucleoli were not visible in 18% of cases, small in 36%, and prominent in 45%. With a mean follow-up of 13.8 months for those without progression (25.9 months' overall follow-up), a repeat biopsy diagnosis of cancer was seen in 10 of 22 men [by report: Gleason score (Gs) 4 (n = 1); Gs 6 (n = 3); Gs 7 (n = 4); Gs 9 (n = 2); three biopsy specimens showed ductal features]. An additional two men developed bone metastases without biopsy. Overall, 12 of 22 (55%) patients had cancer on follow-up (one patient lost to follow-up). Four clinicopathologic findings predicted carcinoma on follow-up: positive DRE (p = 0.02); positive transrectal ultrasound (p = 0.02); bilateral atypical cribriform glands (p = 0.02); and detached cribriform glands (p = 0.04).

Predicting cancer following a diagnosis of high-grade prostatic intraepithelial neoplasia on needle biopsy: data on men with more than one follow-up biopsy.

Most studies on the risk of cancer after high-grade prostatic intraepithelial neoplasia (PIN) on biopsy have been small (fewer than 100 men), have not analyzed in detail if histologic features can predict cancer, and have not assessed the risk of cancer with multiple repeat biopsies. We analyzed 245 men in whom the only abnormal finding on the initial biopsy was high-grade PIN and who had at least one follow-up biopsy. Repeat biopsy identified cancer in 32.2% of men. If only one follow-up biopsy had been performed on the 245 men, only 24.5% of men would have been found to have cancer. The only independent histologic predictor of a cancer diagnosis was the number of cores with high-grade PIN; risk of cancer: 30.2% with 1 or 2 cores, 40% with 3 cores, and 75% with >3 cores. The following did not predict cancer: number of high-grade PIN glands, maximum percentage of gland involved by high-grade PIN, nucleolar prominence, percentage of cells with prominent nucleoli, pattern of high-grade PIN (flat, tufting, micropapillary, cribriform), marked pleomorphism, digital rectal examination, transrectal ultrasound findings, family history of prostate cancer, serum prostate specific antigen (PSA) at time of high-grade PIN diagnosis, and rate of change of serum PSA. Eighty-one (33%) men had more than one follow-up biopsy; in these cases the following findings on the original high-grade PIN biopsy predicted cancer: the presence of mitoses, the number of positive cores, predominant micropapillary and cribriform high-grade PIN, and very large prominent nucleoli. Of 81 men with more than one follow-up biopsy, if the first repeat biopsy were benign, high-grade PIN, or atypical, the eventual cancer rate was 10%, 25.9%, and 57.1%, respectively (p = 0.002). Of 15 men with more than two repeat biopsies, only two (13.3%) had cancer. In summary, approximately one third of men with high-grade PIN on biopsy have cancer on follow-up. If cancer is not found on the first two follow-up biopsies, it will unlikely be found. Although clinical findings at the time of diagnosis of high-grade PIN are not useful to predict who might have cancer, histologic findings may help identify who needs additional biopsies.

Concentrations of oestradiol-17beta and progesterone in the peripheral plasma of the cynomolgus monkey (Macaca fascicularis) in relation to the length of the menstrual cycle and its component phases.

Ninety-three female cynomolgus monkeys were monitored throughout 647 menstrual cycles; 93.2% of these cycles were 22--37 days long and were normally distributed, with a mean of 29.2 days. Menstrual bleeding generally lasted for 3--5 days and was not related to the length of the cycle. The levels of progesterone and oestradiol-17beta in the plasma were measured during the menstrual cycle in 30 monkeys. The concentration of oestradiol-17beta reached a mid-cycle peak on day 11 or 12 of the cycle. The interval between the beginning of the cycle and the oestrogen peak was constant; the interval between the oestrogen peak and the end of the cycle increased or decreased with the cycle length. The level of progesterone began to rise at about the time of the oestrogen peak and remained raised for longer periods as the length of the cycle increased. The length of the menstrual cycle, therefore, appeared to be determined by the duration of the increase in the level of progesterone or by the life of the corpus luteum in the luteal phase. The relationship between the lengths of the cycle and the luteal phase can be defined by the expression: cycle length = 12.6 + 0.96 X length of luteal phase (correlation coefficient = 0.875).

High-grade prostatic intraepithelial neoplasia with adjacent small atypical glands on prostate biopsy.

With high-grade prostatic intraepithelial neoplasia with adjacent small atypical glands (PINATYP), the issue is whether the small glands represent budding or tangentially sectioned glands off of adjacent high-grade prostatic intraepithelial neoplasia (PIN) or invasive cancer next to high-grade PIN. The histology and significance of PINATYP on biopsy have not been described. Among 574 cases of high-grade PIN lesions on needle biopsy, we identified 71 cases of PINATYP. Most cases were consultations, and 51 cases were available for histologic review. At least 1 follow-up prostate biopsy was performed in each of 55 cases. Immunohistochemistry for high-molecular-weight cytokeratin (HMWCK) was performed on cases in which material was available. The average patient age at diagnosis was 65.5 years (range, 48 to 103 years). The initial digital rectal examination, transrectal ultrasound, serum prostate-specific antigen (PSA) level, PSA velocity, and family history of prostate cancer did not predict cancer on repeat biopsy. In 39% of cases, high-grade PIN had a predominantly flat pattern, and remaining cases showed a predominance of other patterns (tufting, micropapillary, cribriform). The average number of high-grade PIN glands and adjacent small atypical glands were 11.5 (1 to 60) and 5.3 (1 to 21), respectively. The farthest adjacent small atypical gland averaged 0.12 mm from the high-grade PIN (0.01 mm to 0.4 mm), as measured with an ocular micrometer. The following histologic features did not predict cancer on repeat biopsy: more than 1 core involved by the high-grade PIN; number of high-grade PIN glands; number of small atypical glands; distance of small atypical glands from the high-grade PIN; size and percentage of nucleoli; marked nuclear pleomorphism; and mitoses. Overall, the risk of cancer on repeat biopsy was 46%. Two findings predicted a lower risk of cancer on repeat biopsy: younger age (62.2 years benign v 68.3 years cancer; P =.004) and predominantly flat high-grade PIN (P =.007). In our material, PINATYP appears to be a greater risk factor than high-grade PIN alone in predicting cancer on rebiopsy. Although age and predominant pattern of associated high-grade PIN may be helpful in predicting which men with this lesion will have cancer on rebiopsy, they cannot be used reliably; therefore, all men with PINATYP should undergo repeat biopsy. HUM PATHOL 32:389-395.

Chemical synthesis and bioassay of anordrin and dinordrin I and II.

The chemical synthesis of 2 alpha, 17 alpha-diethynyl-A-nor-5 alpha-androstane-2 beta, 17 beta-diol dipropionate (Anordrin) and the corresponding diacetate is reported. Similarly, the preparation of the 2 alpha, 17 alpha-diethynyl-A-nor-5 alpha-estrane-2 beta, 17 beta-diol, its diacetate and dipropionate (Dinordrin I), along with the corresponding 2 beta-epimer (Dinordrin II) from 17 beta-hydroxy-A-nor-5 alpha=estran-2-one is described. In rat uterotrophic activity bioassay, the slope of ethynylestradiol differed significantly from the slopes of the other three compounds, thus vitiating potency estimates with this reference compound. Dinordrin I was 20 times more potent than Anordrin and considerably more potent then Dinordrin II. The single-dose oral antifertility effect in rats generally paralleled uterotrophic activity. Immediate postovulatory contraceptive effectiveness was assessed in adult cycling female baboons given two doses daily for 4 days. Both Anordrin and Dinordrin I showed antifertility activity worthy of further study. Moreover, a definite luteolytic effect, with depression of both plasma estrogen and progesterone levels, was observed with these two steroids.

Diagnosis of early pregnancy in the baboon.

PIP: The reliability and rapidity of methods utilizing radioimmunoassay ( RIA) for detecting chorionic gonadotropin (CG) and estrogens in plasma, competitive protein binding assay for plasma progestins, and the hemagglutination inhibition test for urinary CG in the diagnosis of early pregnancy was evaluated in baboons. The hemagglutination inhibition test for detection of urinary CG did not give satisfactory results as late as Day 25 of pregnancy. Confirmation of pregnancy could not be established on Days 8 and 12 of pregnancy by RIA of estrogens, progestins, or CG. However, detection of plasma CG by RIA was 96.6% successful by Day 16, though the method was time-consuming. However, the determination of plasma estrogens and progestins by RIA was determined more quickly on Day 16, and gave equally successful results as the RIA for CG.^ieng

Seizure induction by alcohol in patients with epilepsy experience in two hospital clinics.

We surveyed 70 epileptic patients attending a general neurology clinic and 64 patients attending an epilepsy clinic to determine the incidence of alcohol-related seizures. Seven (10%) of the neurology clinic patients and 9 (15%) of the epilepsy clinic patients reported exacerbation of their seizures with alcohol. In the first group, two had been heavy drinkers when under-age, two had features of alcohol dependence, and three had experienced resolution of seizures following cessation of their drinking. In the second group, five drank 4 units/day or more, and one drank more heavily. The importance of alcohol in the causation of these patients' seizures had not previously been appreciated. The relationship of alcoholism to epilepsy has been recognized for many years, but the role of alcohol in the exacerbation of primary epilepsy, and in triggering seizures in epileptic patients is often not recognized. Control of alcohol ingestion is an important factor in the management of epilepsy.

The isolation of spiroplasmas from mosquitoes in Macon County, Alabama.

During the summer months of 1985, 1,298 adult mosquitoes comprising 21 species and 7 genera were collected in Macon County, Alabama. Mosquitoes were collected from four sections of the county with CO2-baited light traps. Spiroplasma cultures were isolated from two pools of 24 and 25 Aedes fulvus pallens, one pool of 22 Anopheles punctipennis and one pool of 7 Culex nigripalpus. Electron microscopic studies of the isolates revealed helical, wall-less cells.

A survey of childhood hearing impairment.

Full text is available as a scanned copy of the original print version.

Incidence of intestinal parasites in children of different socio-economic population of Dacca city.

Stool samples of 1500 children of different socio-economic status of Dacca city was examined for the presence of intestinal parasites. Samples were collected on voluntary basis which may be treated as randomised. Out of these 1500 children, 816 were male and 684 were female. The overall incidence of parasites was 65.80%, the male children slightly prepondering over the female children. Children of age group 6-10 years showed high incidence of almost all the parasites. Children of Basti area showed the highest incidence (about 90%) and most of them harboured multiple parasites. About 27.13% of infants under the age of one year (mostly from Basti area) harboured parasites. Intestinal parasites were observed that both housing and economic status played vital roles in the occurrence of intestinal parasitosis but the housing and environmental conditions were found to be in a stronger position than economic status.