Surprising deficiency of CENP-B binding sites in African green monkey alpha-satellite DNA: implications for CENP-B function at centromeres.

Centromeres of mammalian chromosomes are rich in repetitive DNAs that are packaged into specialized nucleoprotein structures called heterochromatin. In humans, the major centromeric repetitive DNA, alpha-satellite DNA, has been extensively sequenced and shown to contain binding sites for CENP-B, an 80-kDa centromeric autoantigen. The present report reveals that African green monkey (AGM) cells, which contain extensive alpha-satellite arrays at centromeres, appear to lack the well-characterized CENP-B binding site (the CENP-B box). We show that AGM cells express a functional CENP-B homolog that binds to the CENP-B box and is recognized by several independent anti-CENP-B antibodies. However, three independent assays fail to reveal CENP-B binding sites in AGM DNA. Methods used include a gel mobility shift competition assay using purified AGM alpha-satellite, a novel kinetic electrophoretic mobility shift assay competition protocol using bulk genomic DNA, and bulk sequencing of 76 AGM alpha-satellite monomers. Immunofluorescence studies reveal the presence of significant levels of CENP-B antigen dispersed diffusely throughout the nuclei of interphase cells. These experiments reveal a paradox. CENP-B is highly conserved among mammals, yet its DNA binding site is conserved in human and mouse genomes but not in the AGM genome. One interpretation of these findings is that the role of CENP-B may be in the maintenance and/or organization of centromeric satellite DNA arrays rather than a more direct involvement in centromere structure.

Chemotherapeutic management of small bowel adenocarcinoma associated with Crohn's disease.

Four patients with metastatic primary small bowel adenocarcinoma associated with Crohn's disease were successfully treated with low dose combination chemotherapy consisting of 5-fluorouracil, leucovorin and irinotecan with or without gemcitabine. Benefits included prolonged survival, objective responses, response of resistant tumors, downstaging, and a successful secondary complete resection (Ro) with a durable remission.

Analysis of menstrual records of women immunized with anti-hCG vaccines inducing antibodies partially cross-reactive with hLH.

Menstrual data of 13 control subjects and 88 subjects immunized with three beta-hCG-based vaccine formulations were analysed. Immunization did not change the menstrual regularity; bleeding days were normal (3-7 days) and 89% of the menstrual cycles were within the normal range of 22-35 days. Irregular (short or long) cycles were observed in both immunized and control groups. These were, however, unrelated to prevailing anti-hCG antibody titres or to cross-reactivity of antibodies with hLH.

Phase I clinical trials with three formulations of anti-human chorionic gonadotropin vaccine.

Comparative phase I clinical trials were carried out in 5 centres with three formulations of beta-hCG-based vaccines inducing antibodies against human chorionic gonadotropin. The objectives of these trials were to determine their relative immunogenicity, duration, reversibility and safety. A total of 116 tubal ligated women volunteers were enrolled in the study and 101 subjects were followed-up for one year or more until the antibody titres declined to near zero levels. Every woman receiving the vaccine produced anti-hCG and anti-tetanus antibodies. Clinical examination carried out at intervals of 4-6 weeks revealed no abnormality. No serious side effects or adverse reactions were reported with any of the formulations during primary immunization with three monthly injections of the vaccine. Eleven women, however, demonstrated hypersensitivity to test dose at the time of the booster injection. The reaction was to tetanus toxoid; gonadotropin subunits conjugated to another carrier did not evoke any such reaction. Progesterone in bleeds taken at midluteal phase, as well as complete progesterone and estradiol done in two immunized women, indicated normal ovulatory cycles. Immunization with these formulations had no significant effect on haematological, clinical chemistry and other metabolic parameters. In summary, the results indicate that none of the three beta-hCG-based contraceptive vaccines had any adverse effects clinically, on endocrine status and metabolic parameters. Formulations A and B induced comparatively higher anti-hCG titres than M. Thus, further work can be undertaken to study the efficacy of these vaccines in humans for preventing pregnancy.

Concurrent copper T insertion with medical termination of pregnancy in women with previous caesarean section delivery.

PIP: The event rates up to 1 year after insertion of a Copper-T IUD were compared in 76 women who had 1 or more cesarean sections, and were given their IUD immediately after medical termination of pregnancy, and in 76 women matched for age and parity but normal vaginal deliveries. The cesarean group had abortion performed under general anesthesia; the vaginal group had iv diazepam and paracervical block. All subjects were less than 11 weeks gestation. They were followed up at 7 days, 6 weeks, and 3 monthly intervals for 1 year. No perforations or pregnancies occurred. Incomplete abortion caused expulsion in 2.6% of women in both groups, and removal in 6.5 and 5.3%, in the cesarean and control groups respectively, and was responsible for most discontinuations. It was concluded that IUD insertion is safe after medical termination of pregnancy in women with a history of cesarean section, depending on the skill of the surgeon.^ieng

Spectrophotometric analysis of amniotic fluid in Rh immunised pregnancies.

A total of 307 amniotic fluid analysis done in 344 Rh negative immunised mothers showed that 46, 83 and 178 delta OD values at 450 millimicrons fell in the upper, middle and lower zones of Liley's charts respectively. The correlation of spectrophotometric analysis with the condition of the baby at birth was about 95 per cent in the upper and lower zones. In the middle zone, however, it was about 75 per cent only. Also, in 7 women in whom the OD at 450 millimicrons fell in the middle zone, the babies were found to be Rh negative; in another baby, the OD difference fell in upper zone. In spite of these limitations amniotic fluid examination seems to be an important single guide to severity, being superior to other parameters like previous obstetric history, antibody titre alone and Liley's charts, which are still widely used.

Role of the male partner in the lower genitourinary tract infection of female.

We studied the relationships of selected microbes and the role of consorts in the causation of vaginal discharge which may be due to cervicitis or vaginitis. A total of 93 consecutive patients in the reproductive age group with symptoms of vaginal discharge along with their sexual partners were studied. Samples were collected from the cervix and posterior fornix of the female patients and from the urethra and sub-prepucial area of the male partner for culture of Neisseria gonorrhoeae, Gardnerella vaginalis, Mycoplasma hominis, ureaplasma, candida, aerobic and anaerobic organisms. Apart from cultures, KOH and Gram stain of the discharge were made. Predominant pathogen isolated was Ureaplasma urealyticum from 40 (43.01%) females and 23 (24.7%) males. The next common pathogenic organisms isolated were candida species from 11 (11.8%) females and 5 (5.4%) males and Chlamydia trachomatis in 3 (3.2%) females and 1 (1.1%) male. Various organisms were more frequently isolated from the 29 of 43 (67.4%) couples who had had sexual intercourse 7 days prior to the recruitment as compared to 14 of 43 (32.6%) who had had coitus more than 7 days prior to their recruitment. This may be due to the spontaneous disappearance or decrease in the number of organisms to the level that they could be detected by culture. In our study, 6 (6.5%) of male partners carrying pathogenic organisms were asymptomatic, indicating that their screening and treatment is vital.

Oxytocin induction of labor: a comparison of 20- and 60-min dose increment levels.

OBJECTIVES: To compare the efficacy and complications of oxytocin dose increments at 20- and 60-min intervals for induction of labor in women with low parity. METHODS: One hundred women of low parity requiring induction of labor were randomly allocated to 20- and 60-min oxytocin dose increments, 50 patients in each group. The basal oxytocin dose was 1 milliunit/min and doubling of the oxytocin dose was done at intervals of 20 and 60 min. RESULTS: The group with 60-min increments had a decreased incidence of uterine hyperstimulation, cesarean section and operative vaginal delivery. The induction-delivery interval was similar in both groups. CONCLUSION: The oxytocin infusion regimen with increments at 60-min intervals is safer than and equally effective as 20-min incremental intervals.

Safety and efficacy of stripping of membranes at term.

OBJECTIVES: To assess the efficacy of stripping of membranes in initiation of labor and to study its effect on maternal and perinatal morbidity. METHOD: One-hundred primigravidae with certain gestational dates were randomized at 38 weeks gestation to either receive stripping of membranes or only gentle cervical examination. Cervical swabs were taken before pelvic examination at 38 weeks and again at the onset of labor. Placental membranes were sent for bacteriological study after delivery in all patients. RESULTS: The mean gestational age, parity and Bishop score were similar in both groups at recruitment. Gestational age at delivery was lower in the study group (38.70 +/- 0.63) compared to the control group. Seventy-two percent of the study group and 8% of the control group had spontaneous onset of labor within 7 days of examination. Labor was induced in one patient (2%) of the study group and 16 patients (32%) of the control group. No statistically significant difference was noted in incidence of premature rupture of membranes (PROM), mode of delivery, intrapartum events and perinatal outcome. No increase in neonatal morbidity was seen in association with this procedure. No patient in the study group had clinical evidence of chorioamnionitis. There was no statistically significant difference in the microbiological flora of both groups. CONCLUSION: Stripping of the fetal membranes is a safe and efficacious procedure for induction of labor. It decreases the incidence of induction of labor with no increase in incidence of maternal and neonatal morbidity.

Pregnancy following cardiac valve replacement surgery.

OBJECTIVES: To study the outcome of pregnancy in women with artificial heart valves and to compare the maternal and perinatal outcome in mechanical and bioprosthetic valves. METHOD: Retrospective analysis of 34 pregnancies in 29 women who conceived after cardiac valve replacement was carried out. RESULTS: The majority of women (76.4%) delivered within 5 years of valve replacement. Anticoagulants were administered in 79.4% of pregnancies. Maternal mortality was 2.9% and maternal morbidity in the form of heart failure, atrial fibrillation, valve thrombosis, thromboembolism, bleeding complications and non-functioning prostheses were 2.9%, 5.8%, 2.9%, 2.9%, 11.7% and 2.9%, respectively. The incidence of prematurity was 5.8% and small for gestational age babies was 11.7%. There was no case of abortion. Two babies (5.8%) were still born, one of which had malformations. Maternal complications were significantly higher in women with bioprostheses, though the complications were more grave in the mechanical prostheses group. The perinatal outcome was almost similar in both the groups. CONCLUSION: The perinatal outcome was not different in women with bioprosthetic valves from the ones with mechanical prostheses, but the maternal morbidity was more in women with bioprosthetic valves. Coumarin derivatives were safe and effective and did not lead to embryopathy.

Calcium metabolism in pre-eclampsia.

OBJECTIVES: To study calcium metabolism in pre-eclampsia and normotensive gravid women. METHOD: Ten milliliters of heparinized blood samples and 24-h urine samples were collected from 50 pre-eclamptic and 50 normotensive primigravidae. Blood samples were studied for calcium uptake, intracellular calcium level and calcium-dependent adenosine triphosphatase activity of red blood cell ghost. Urinary calcium excretion was estimated from the 24-h urine samples. These values were compared in the two groups. RESULTS: The mean gestational age at recruitment was similar in both the groups. The mean maternal age was 24.28 +/- 2.41 years in pre-eclamptic and 23.48 +/- 4.16 years in normotensive women. In pre-eclampsia 24-h urinary calcium excretion (71.20 +/- 22.95 mg/day) and calcium-dependent ATPase activity (10.78 +/- 2.40 nmol/Pi/mg protein/min) was significantly lower compared to normotensive primigravidae (calcium excretion = 189.24 +/- 57.06 mg/day; Ca2+-dependent ATPase = 12.64 +/- 2.42 nmolPi/mg /protein per min; P < 0.001). Intracellular calcium levels and calcium uptake at 10 min by red blood cells were significantly higher in pre-eclampsia (P < 0.05). Calcium uptake by red blood cells at 20 and 30 min was similar in both groups. CONCLUSION: Pre-eclampsia is associated with increased levels of intracellular calcium, decreased calcium-dependent ATPase activity of erythrocytes and hypocalciuria.

Non-cirrhotic portal hypertension in pregnancy.

OBJECTIVE: To study the outcome of pregnancy in women with non-cirrhotic portal hypertension (NCPH). METHOD: A retrospective analysis of 50 pregnancies in 27 women with NCPH was carried out. Pregnancy outcome was compared in extra hepatic portal vein obstruction (EHPVO) and non-cirrhotic portal fibrosis (NCPF). RESULTS: The mean maternal age was 24.60+/-2.857 years, and the disease was diagnosed during pregnancy in 15 (55.6%) patients. Variceal bleeding occurred in 17/50 (34%) pregnancies and the majority (88.2%) of them responded to endoscopic sclerotherapy. Incidence of variceal bleeding during pregnancy was lower in pregnancies where the disease was diagnosed prior to pregnancy (8.6%), and it was 43.5% in EHPVO and 25.9% in NCPF. The mean birth weight of the neonates was 2668.4+/-427.42 g, and the incidence of abortion, prematurity, small for gestational age babies and perinatal death was 20, 17.5, 12.5 and 20%, respectively. Variceal bleeding during pregnancy was associated with a higher incidence of abortion (29.4%) and perinatal death (33.3%). CONCLUSION: Variceal bleeding is the most common complication in pregnancies with NCPH. Pregnancies can be allowed and managed successfully in patients with NCPH.

Pregnancy with epilepsy--a retrospective analysis.

OBJECTIVES: To study the course of epilepsy in pregnancy and to assess the perinatal outcome. METHODS: A retrospective analysis of 219 pregnant patients with epilepsy. The type of epilepsy, drug therapy and seizure frequency were documented. The perinatal outcome of 157 pregnancies with epilepsy was analyzed and compared with that of 471 normal gravidas of similar age and parity. RESULTS: Generalized seizures occurred in 203 patients, partial seizures in 13 patients and complex partial seizures in three patients. One hundred fifty-two patients (69.41%) were on monotherapy. Carbamazepine was the most common drug (56.58%) used. Ninety-five patients (43.38%) had seizures in the current pregnancy, five of whom had status epilepticus. There was no maternal mortality in status epilepticus. There was no difference in perinatal outcome between the study and control groups. The incidence of congenital malformations was higher in the control group (5/476, 1.05%) than in the study group (1/160, 0.63%). The incidence of low-birth-weight babies was higher in the study group in patients with gestational seizures. CONCLUSION: The course of pregnancy and perinatal outcome was not altered by epilepsy. There was no increase in the incidence of congenital malformations with the use of monotherapy.

Maternal and perinatal outcome in rheumatic heart disease.

OBJECTIVES: To study the maternal and perinatal outcome of pregnancies complicated by rheumatic heart disease. METHODS: A retrospective study was carried out in the cardio-obstetric clinic of the Postgraduate Institute of Medical Education and Research, Chandigarh (India) over a period of 13 years (1987-1999) involving 486 pregnant patients with rheumatic heart disease. Maternal and perinatal outcome was reviewed. RESULTS: Three hundred and four patients (63.3%) had single valve involvement and mitral stenosis was the most predominant lesion (89.2%). One hundred and seventy one (38.6%) patients had undergone surgical correction prior to the onset of pregnancy. One hundred and thirteen patients (22.6%) were identified as NYHA class III-IV. Mitral valvotomy was performed during pregnancy in 48 patients. The incidence of preterm birth and small for gestational age newborns was 12% and 18.2%, respectively. There were 10 maternal deaths, of which eight patients were NYHA III and IV. CONCLUSIONS: Rheumatic heart disease in pregnancy is associated with significant maternal and perinatal morbidity in NYHA class III-IV patients.

Predilection of hemolytic anemia-associated leg ulcers for the medial malleolus.

BACKGROUND: Ischemic ulcers are usually found above the lateral, and venous stasis ulcers at the medial malleoli. Leg ulcers occur in at least 25% of sickle cell disease (SSD) patients in clinic populations, usually in the malleolar region. The function of the large leg veins in most SSD patients is unimpaired. PATIENTS AND METHODS: We determined leg ulcer location in 41 sickle cell anemia (SS), and 4 sickle-beta 0 thalassemic patients with longstanding chronic and/or recurrent leg ulceration, and reviewed published reports of leg ulcers in hereditary spherocytosis and thalassemias. RESULTS: Of the 57 legs of the 45 SSD patients with only 1 ulcer, 42 (74%) were medial and 15 lateral. The difference was significant (p < 0.001). Of patients with only a single ulcer, 22 were medial and 4 lateral. Of 15 reported patients with leg ulcers related to spherocytosis or thalassemia, 20/24 (83%) ulcers were medial. CONCLUSIONS: The medial malleoli are the most common site of leg ulceration in SSD and in other chronic hemolytic anemias. This suggests that stasis may play a role in the leg ulceration associated with chronic hemolytic anemia.

Neurological complications of eclampsia.

OBJECTIVE: A prospective study was conducted to evaluate the various neurological (clinical, radiological and EEG) complications in patients of eclampsia. METHODS: Thirty nine patients of eclampsia were studied regarding neurological findings at presentation and electroencephalographic (EEG) tracings were recorded in each patient. Patients with an abnormal neurologic examination and/or focal or lateralizing findings on EEG, underwent a CT scan (n = 18). Foetal and maternal outcome were recorded. RESULTS: The age of the patients ranged from 19-30 (mean +/- SD, 24.2 +/- 3.5) years thirty six patients (92%) had seizures in the antenatal period, 2 (5.4%) patients developed post partum eclampsia and 1 (2.6%) patient had seizures before and after delivery. A diffuse encephalopathy was seen in 9 patients (23.1%), 4 patients (10.2%) had hemiparesis and 1 patient (2.6%) had papilledema. EEG abnormalities were seen in 29 cases (74%) and included generalized slowing (n = 19), generalized sharp waves (n = 9), focal slowing (n = 4), focal sharp waves (n = 2) and spikes (generalized and focal) were seen in 1 patient each. Abnormal CT scan was seen in 10 cases (n = 18). Five patients had generalized infarct was seen in 1 patient each. There were 8 (20.5%) still births and 31 (19.5%) live births and no maternal mortality. CONCLUSIONS: Antenatal seizures occur in > 90% cases of eclampsia and less than 10% cases have seizures after delivery. A diffuse encephalopathy is the commonest clinical abnormality along with generalized slowing on EEG. Although cerebral oedema is common focal infarcts may be seen on CT scan.

3D optic disc reconstruction via a global fundus stereo algorithm.

This paper presents a novel method to recover 3D structure of the optic disc in the retina from two uncalibrated fundus images. Retinal images are commonly uncalibrated when acquired clinically, creating rectification challenges as well as significant radiometric and blur differences within the stereo pair. By exploiting structural peculiarities of the retina, we modified the Graph Cuts computational stereo method (one of current state-of-the-art methods) to yield a high quality algorithm for fundus stereo reconstruction. Extensive qualitative and quantitative experimental evaluation (where OCT scans are used as 3D ground truth) on our and publicly available datasets shows the superiority of the proposed method in comparison to other alternatives.

Laparoscopic removal of translocated intrauterine contraceptive devices.

Data is presented regarding 20 translocated intrauterine contraceptive devices (IUCD). Successful laparoscopic removal was made in 13 (65%) cases though all these devices were Copper 'T' 200 model which tend to produce dense adhesions. Only when the IUCD was suspected to be partially or fully in the gut lumen or thick adhesions were present, or the device was not visualised, was laparotomy performed. Hence it is advocated that laparoscopy should be performed as a routine in patients with suspected translocated IUCD, as it obviates the need for laparotomy and thus decreases the duration of the hospital stay as well as preventing morbidity associated with laparotomy.

PIP: Data is presented regarding 20 translocated IUDs. Successful laparoscopic removal was made in 13 (65%) cases although these devices were all Copper T 200 models, which tend to produce dense adhesions. Only when the IUD was suspected to be partially or fully in the gut lumen or thick adhesions were present, or the device was not visualized, was laparotomy performed. Thus, it is advocated that laparoscopy by performed as a routine procedure in patients with suspected translocated IUD, as it obviates the need for laparotomy and decreases the duration of the hospital. It furthermore prevents morbidity associated with laparotomy.