Evaluation of pharmacokinetics of single-dose primaquine in undernourished versus normally nourished children diagnosed with Plasmodium vivax malaria in Mumbai.

BACKGROUND: The pharmacokinetics of primaquine [PQ] have been the subject of studies in both adults and healthy participants. However, there is no study on its pharmacokinetics in a setting of undernourishment. In India, there is evidence to show considerable malnourishment in children that in turn can affect drug pharmacokinetics. Given that the country is moving towards malaria elimination, the present study was planned with the objective of comparing pharmacokinetics of the drug in undernourished children relative to normally nourished children. MATERIALS AND METHODS: After Institutional Ethics Committee approval, children of either gender between the ages of 5 and 12 years and smear-positive for Plasmodium vivax malaria were included. Nourishment status was determined using the Indian Academy of Pediatrics classification of protein energy malnutrition based on Khadilkar's growth charts. Twelve children each were enrolled in the two groups. PQ was given in the dose of 0.3 mg/kg/d and blood collections were made at 0, 1, 2, 3, 4, 6, 8 and 24 hours post-dosing. Levels were estimated by high-performance liquid chromatography. Chloroquine in the dose of 25 mg/kg was given over three days along with supportive care. RESULTS: Of the 24 children, there were 17 boys and 7 girls. There was a statistically significant difference in the body weight between the undernourished and the normally nourished children [21.5 ± 5.52 vs. 28.8 ± 8.84, P < 0.05]. PQ levels showed wide inter-individual variation in both groups. No significant difference was seen in any pharmacokinetic parameter between the two groups. DISCUSSION: This study adds to the limited body of evidence on the pharmacokinetics of PQ in children with malaria and indicates that the dosing of primaquine could potentially be independent of the nourishment status.

Complete spermatogenesis in intratesticular testis tissue xenotransplants from immature non-human primate.

STUDY QUESTION: Can full spermatogenesis be achieved after xenotransplantation of prepubertal primate testis tissue to the mouse, in testis or subcutaneously? SUMMARY ANSWER: Intratesticular xenotransplantation supported the differentiation of immature germ cells from marmoset (Callithrix jacchus) into spermatids and spermatozoa at 4 and 9 months post-transplantation, while in subcutaneous transplants, spermatogenic arrest was observed at 4 months and none of the transplants survived at 9 months. WHAT IS KNOWN ALREADY: Auto-transplantation of cryopreserved immature testis tissue (ITT) could be a potential fertility restoration strategy for patients with complete loss of germ cells due to chemo- and/or radiotherapy at a young age. Before ITT transplantation can be used for clinical application, it is a prerequisite to demonstrate the feasibility of the technique and identify the conditions required for establishing spermatogenesis in primate ITT transplants. Although xenotransplantation of ITT from several species has resulted in complete spermatogenesis, in human and marmoset, ITT has not been successful. STUDY DESIGN, SIZE, DURATION: In this study, we used marmoset as a pre-clinical animal model. ITT was obtained from two 6-month-old co-twin marmosets. A total of 147 testis tissue pieces (~0.8-1.0 mm3 each) were transplanted into the testicular parenchyma (intratesticular; n = 40) or under the dorsal skin (ectopic; n = 107) of 4-week-old immunodeficient Swiss Nu/Nu mice (n = 20). Each mouse received one single marmoset testis tissue piece in each testis and 4-6 pieces subcutaneously. Xenotransplants were retrieved at 4 and 9 months post-transplantation and evaluations were performed with regards to transplant survival, spermatogonial quantity and germ cell differentiation. PARTICIPANTS/MATERIALS, SETTING, METHODS: Transplant survival was histologically evaluated by haematoxylin-periodic acid Schiff (H/PAS) staining. Spermatogonia were identified by MAGE-A4 via immunohistochemistry. Germ cell differentiation was assessed by morphological identification of different germ cell types on H/PAS stained sections. Meiotically active germ cells were identified by BOLL expression. CREM immunohistochemistry was performed to confirm the presence of post-meiotic germ cells and ACROSIN was used to determine the presence of round, elongating and elongated spermatids. MAIN RESULTS AND THE ROLE OF CHANCE: Four months post-transplantation, 50% of the intratesticular transplants and 21% of the ectopic transplants were recovered (P = 0.019). The number of spermatogonia per tubule did not show any variation. In 33% of the recovered intratesticular transplants, complete spermatogenesis was established. Overall, 78% of the intratesticular transplants showed post-meiotic differentiation (round spermatids, elongating/elongated spermatids and spermatozoa). However, during the same period, spermatocytes (early meiotic germ cells) were the most advanced germ cell type present in the ectopic transplants. Nine months post-transplantation, 50% of the intratesticular transplants survived, whilst none of the ectopic transplants was recovered (P < 0.0001). Transplants contained more spermatogonia per tubule (P = 0.018) than at 4 months. Complete spermatogenesis was observed in all recovered transplants (100%), indicating a progressive spermatogenic development in intratesticular transplants between the two time-points. Nine months post-transplantation, transplants contained more seminiferous tubules with post-meiotic germ cells (37 vs. 5%; P < 0.001) and fewer tubules without germ cells (2 vs. 8%; P = 0.014) compared to 4 months post-transplantation. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Although xenotransplantation of marmoset ITT was successful, it does not fully reflect all aspects of a future clinical setting. Furthermore, due to ethical restrictions, we were not able to prove the functionality of the spermatozoa produced in the marmoset transplants. WIDER IMPLICATIONS OF THE FINDINGS: In this pre-clinical study, we demonstrated that testicular parenchyma provides the required microenvironment for germ cell differentiation and long-term survival of immature marmoset testis tissue, likely due to the favourable temperature regulation, growth factors and hormonal support. These results encourage the design of new experiments on human ITT xenotransplantation and show that intratesticular transplantation is likely to be superior to ectopic transplantation for fertility restoration following gonadotoxic treatment in childhood. STUDY FUNDING/COMPETING INTEREST(S): This project was funded by the ITN Marie Curie Programme 'Growsperm' (EU-FP7-PEOPLE-2013-ITN 603568) and the scientific Fund Willy Gepts from the UZ Brussel (ADSI677). D.V.S. is a post-doctoral fellow of the Fonds Wetenschappelijk Onderzoek (FWO; 12M2815N). No conflict of interest is declared.

Formulation development and evaluation of antimicrobial polyherbal gel.

OBJECTIVE: In the recent years, there has been a gradual revival of interest in the use of medicinal plants in developing countries because herbal medicines have been reported safe with minimal adverse side effect especially when compared with synthetic drugs. METHOD: In the present study we prepared gel formulations (formulations A and B) which comprised of ethanolic extract of Azadirachta indica, Curcuma longa, Allium sativum, Ocimum sactum, Cinnamomum zeylanicum nees and Tamarindus indica in a concentration of 0.1 and 0.5%, respectively in a base. The base was prepared by using carbapol 940, propylene glycol-400, ethanol, methyl paraben, propylparaben, EDTA, triethanolamine and required amount of water in a quantity sufficient to prepare 50g. The prepared formulations were screened for their antimicrobial activity by agar well diffusion technique against S. aureus, B. subtilis, A. niger and E. coli which are representative types of Gram positive and Gram negative organisms. The formulations were also evaluated for appearance and homogeneity, pH, viscosity and rheological studies, spreadability, drug content uniformity, skin irritation test (Patch test) and washability. RESULT: The results of the studies revealed that both formulation under study viz A and B showed better zone of inhibition as compared with the base. However, formulation B exhibited maximum activity against the selected strains which may be attributed to its greater amount of herbal extracts as compared to formulation A. CONCLUSION: Based on our research, it could be concluded that these formulations possess antimicrobial activity and can be used safely on human skin.

Can mesenchymal stem cells improve spermatogonial stem cell transplantation efficiency?

Improved treatments have led to an increased survival rate in cancer patients. However, in pre-pubertal boys, these gonadotoxic treatments can result in the depletion of the spermatogonial stem cell (SSC) pool causing lifelong infertility. SSC transplantation has been proposed as a promising technique to preserve the fertility of these patients. In mice, this technique has resulted in live-born offspring, but the efficiency of colonization remained low. This could be because of a deficient microenvironment, leading to apoptosis of the transplanted SSCs. Interestingly, mesenchymal stem cells (MSCs), being multipotent and easy to isolate and multiply in vitro, are nowadays successfully and widely used in regenerative medicine. Here, we shortly review the current understanding of MSC and SSC biology, and we hypothesize that a combined MSC-SSC transplantation might improve the efficiency of SSC colonization and differentiation as paracrine factors from MSCs may contribute to the SSC niche.

Genetic determinants of lipid-lowering response to atorvastatin therapy in an Indian population.

WHAT IS KNOWN AND OBJECTIVE: Statins form the backbone of lipid-lowering therapy for the prevention of cardiovascular disease. However, there is large interindividual variability in clinical response to statin treatment. Several gene variants that can be aligned to either the pharmacokinetics or pharmacodynamics of statin have been proposed as potentially important determinants of statin response. We aimed to study the association of known variations in SLCO1B1, CYP3A4, ABCB1, CYP3A5, ABCG5 and CYP7A1 genes with lipid levels in response to atorvastatin therapy. METHODS: Genotypes were determined using multiplex allele-specific polymerase chain reaction in 177 Indian patients, treated with 10 mg of atorvastatin for 8 weeks. Low-density lipoprotein-cholesterol (LDL-C) levels were recorded at baseline and after 8 weeks of atorvastatin treatment. RESULTS AND DISCUSSION: A total of 177 hypercholesterolaemic patients were genotyped to study genetic determinants of atorvastatin response. The genotype distribution for all polymorphisms investigated was in Hardy-Weinberg equilibrium. In our study, patients with wild-type genotypes of CYP7A1 (rs3808607), CYP3A4 (rs2740574), SLCO1B1 (rs2306283) and variant allele-carrying genotype of ABCB1 (rs2032582, rs1045642) showed significantly greater LDL-cholesterol reductions in response to atorvastatin therapy. WHAT IS NEW AND CONCLUSION: The variable response to atorvastatin therapy in terms of LDL-cholesterol lowering due to genetic variations in CYP7A1, CYP3A4, SLCO1B1 and ABCB1 is a promising finding. Further validation in large Indian cohorts is required before it can be assessed for clinical utility.

Anesthetic Management of a Patient with DRESS Syndrome for Renal Transplantation.

Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome is a rare but potentially life-threatening hypersensitivity reaction characterized by widespread erythematous skin eruptions with fever, lymphadenopathy and visceral involvement-hepatitis, nephritis, pericarditis, and pneumonitis. There are numerous reports describing the management of such patients in intensive care units but hardly any describing the intraoperative anesthetic management of such patients. Herein, we report on a patient with DRESS syndrome who was scheduled for renal transplantation. The main goal in this case was to prevent a hypersensitive drug reaction intraoperatively and develop a safe alternative anesthesia plan for the patient. After pre-operative skin and intradermal tests, we chose the drugs that could be safely used for anesthesia. Usually general anesthesia is preferred for renal transplantation but in this patient we opted for combined spinal epidural anesthesia. Precautions that are to taken in such a case and the anesthetic management are discussed in detail herewith.

Immunoglobulin levels and CD4 / CD8 counts in ß- thalassemia major.

OBJECTIVE: This cross-sectional study determined the CD4, CD8 counts and serum immunoglobulins in transfusion dependent b - thalassemic patients, and correlated them with anti-HIV, anti-HCV and HBsAg status, number of transfusions, iron overload and splenectomy. METHODS: Patients with acute or chronic diseases (except HIV, Hepatitis B and C), on immunosuppressive drugs or vaccinated within one month prior to study were excluded. CD4, CD8 counts and serum Immunoglobulins were documented. RESULTS: Increasing transfusions led to higher IgA and IgM as well as a decline in CD4 and CD8 levels. Higher ferritin correlated with high IgM. CD4, CD8 and IgA were significantly higher in splenectomized subjects. HCV correlated significantly with lower IgA values. CONCLUSION: Higher transfusion requirement, iron overload, splenectomy and HCV infection correlated with alterations in different immunological parameters.

Intercity deceased donor renal transplantation: a single-center experience from a developing country.

In a developing country such as India, deceased donor renal transplantation (DDRTx) accounts for only about 1% of all renal transplants (RTx). Our institute initiated an intercity DDRTx in the year 2006, which significantly increased the number of RTx. We retrieved 74 kidneys from 37 deceased donors from various cities of Gujarat from January 2006 to December 2009. We transplanted the allografts in 66 recipients and a retrospective analysis of the donor profile and management and recipient outcome was performed. The mean age of the donors was 43.3 ± 18.8 years. The causes of death included road traffic accident in 51.35% of the donors and cerebrovascular stroke in 48.65% of the donors; 83.78% of the donors required ionotropes for hemodynamic stability in addition to vigorous intravenous fluid replacement. The average urine output of the donors was 350 ± 150 mL. The organs were perfused and stored in HTK solution. The mean cold ischemia time (CIT) was 9.12 ± 5.25 h. The mean anastomosis time in the recipient was 30.8 ± 8.7 min. 57.6% of the recipients established urine output on the operating table and 42.4% developed delayed graft function. At the end of 1 month after transplantation, the mean serum creatinine was comparable to the Ahmadabad city DDRTx, although the CIT was significantly longer in the intercity patients. Intercity organ harvesting is a viable option to increase the donor pool. Distance may not be an impediment, and good recipient outcome is possible in spite of prolonged CIT in case of proper harvesting and preservation.

Is there a role for metronomic induction (and maintenance) therapy in elderly patients with acute myeloid leukemia? A literature review.

Acute myeloid leukemia (AML) in older adults differs biologically and clinically from that in younger patients and is characterized by adverse chromosomal abnormalities, stronger intrinsic resistance, and lower tolerance to chemotherapy. In patients over age 60 with AML, cure rates are under 10% despite intensive chemotherapy, and most of them die within a year of diagnosis. Over the last decade, metronomic chemotherapy has emerged as a potential strategy to control advanced/refractory cancer. Here, we report a case of a 68-year-old gentleman having AML with high-risk cytogenetic features, who achieved complete remission on our oral metronomic PrET (PrET: Prednisolone, etoposide, thioguanine) protocol on an outpatient basis. He was later treated with standard high-dose (HD) cytosine arabinoside (Ara-C) consolidation followed by maintenance with etoposide, thioguanine, and sodium valproate. Presently, the patient is nearly 35 months since diagnosis and 21 months off treatment. This case report and review highlights that the combination of oral low-intensity metronomic therapy, followed by standard HD consolidation therapy and metronomic maintenance therapy may be well tolerated by elderly patients especially with less proliferative, high (cytogenetic)-risk AML who are otherwise deemed to be unfit for intensive intravenous induction chemotherapy regimens. References for this review were identified through searches of Pubmed for recent publications on the subject as well as searches of the files of the authors themselves. The final list was generated on the basis of originality and relevance to this review.

In vitro culture and morphological characterization of prepubertal buffalo (Bubalus bubalis) putative spermatogonial stem cell.

PURPOSE: Spermatogonial stem cells (SSCs) have the unique ability both to self-renew and to produce progeny that undergo differentiation to spermatozoa. The present study has been carried out to develop a method to purify and enrich the pure populations of spermatogonial stem cell like cells in buffalo. METHODS: The spermatogonial cells were isolated from testes of 3-7 month old buffalo calves and disaggregated by double enzymatic digestion. Mixed population of isolated cells were then plated on Datura stramonium agglutinin (DSA) lectin coated dishes for attachment of Sertoli cells. The desired cells were obtained from suspension medium after 18 h of incubation and then loaded on discontinuous density gradient using percoll (20-65 %) and different types of spermatogonia cells were obtained at interface of each layer. These cells were cultured in vitro. RESULTS: Spermatogonial cells isolated have spherical outline and two or three eccentrically placed nucleoli, created a colony after proliferation during first week or immediately after passage. After 7-10 days of culture, the resulted developed colonies of spermatogonial cells expressed the spermatogonial specific genes like Plzf and VASA; and other pluripotency related markers viz. alkaline phosphtase, DBA, CD9, CD90, SSEA-1, OCT-4, NANOG and REX-1. CONCLUSION: Our results show that the isolated putative spermatogonial stem cells exhibit the expression of pluripotency related and spermatogonial specific genes. This study may help to establish a long term culture system for buffalo spermatogonia.

Protective effect of Luffa acutangula extracts on gastric ulceration in NIDDM rats: role of gastric mucosal glycoproteins and antioxidants.

OBJECTIVE: To study the comparative gastroprotective effect of Luffa acutangula methanolic extract (LAM) and aqueous extract (LAW) on type II diabetes rats. METHODS: Streptozotocin (65 mg/kg, i.p.) along with nicotinamide (120 mg/kg, i.p.) was used to induce non insulin dependent diabetes mellitus (NIDDM) in rats. A daily oral dose of aspirin (200 mg/kg, i.p.) was administered for initial seven days to induce gastric ulcerations in the diabetic rats. LAM and LAW were administered orally in the doses of 100, 200 and 400 mg/kg once daily for 21 days. Glibenclamide and ranitidine were used as standards for comparing the antidiabetic and antiulcer effect respectively. RESULTS: LAM significantly (P<0.01) increased mucosal glycoprotein and antioxidant enzyme level in gastric mucosa of diabetic rats than LAW (P <0.05). LAM was efficient in reversing the delayed healing of gastric ulcer in diabetic rats close to the normal level. LAM exhibited better ulcer healing effect than glibenclamide and LAW, because of its both antihyperglycemic and mucosal defensive actions. CONCLUSIONS: Thus, LAM is proved to be a better alternative for treating gastric ulcers co-occurring with diabetes.

Immunophenotypic profile of acute leukemia: critical analysis and insights gained at a tertiary care center in India.

BACKGROUND: To analyze the spectrum of various types and subtypes of acute leukemia. METHODS: Two thousand five hundred and eleven consecutive new referral cases of acute leukemia (AL) were evaluated based on WHO classification. RESULTS: It included 1,471 cases (58%) of acute lymphoblastic leukemia (ALL), 964 cases (38%) of acute myeloid leukemia (AML), 45 cases (1.8%) of chronic myelogenous leukemia in blast crisis (CMLBC), 37 cases (1.5%) of biphenotypic acute leukemia (BAL), 1 case of Triphenotypic AL, and 2 cases of acute undifferentiated leukemia (AUL). Common subtypes of ALL were B-cell ALL (76%), which comprised of intermediate stage/CALLA positive (73%), early precursor/proBALL (3%). T-cell ALL constituted 24% (351 cases) of ALL. Common subtypes of AML included AMLM2 (27%), AMLM5 (15%), AMLM0 (12%), AMLM1 (12%), APML (11%), and AML t(8;21) (9%). CMLBC was commonly of myeloid blast crisis subtype (40 cases). CONCLUSION: B-cell ALL was the commonest subtype in children and AML in adults. Overall incidence of AML in adults was low (53% only). CD13 was most sensitive and CD117 most specific for determining myeloid lineage. A minimal primary panel of nine antibodies consisting of three myeloid markers (CD13, CD33, and CD117), B-cell lymphoid marker (CD19), T-cell marker (CD7), with CD45, CD10, CD34, and HLADR could assign lineage to 92% of AL. Cytogenetics findings lead to a change in the diagnostic subtype of myeloid malignancy in 38 (1.5%) cases.

Carbon dioxide absorption during laparoscopic donor nephrectomy: a comparison between retroperitoneal and transperitoneal approaches.

AIM: Previous studies have suggested that retroperitoneal and transperitoneal approaches for laparoscopic donor nephrectomy are associated with variable carbon dioxide (CO(2)) absorption, which can cause significant morbidity. The approach that results in greater CO(2) absorption is a matter of debate. We studied patients undergoing transperitoneal/retroperitoneal donor nephrectomy to determine relative CO(2) absorption, incidence of subcutaneous emphysema, pneumothorax, and pneumomediastinum, seeking to establish a correlation between the incidence of subcutaneous emphysema and CO(2) elimination. MATERIALS AND METHODS: This was a prospective nonrandomized, single-center, two-arm clinical study of 60 kidney donors undergoing laparoscopic nephrectomy by transperitoneal (n = 30) or retroperitoneal (n = 30) approach. CO(2) elimination was calculated using end tidal CO(2), tidal volume, respiratory rate, and weight of the donor. We studied intraoperative CO(2) elimination and CO(2) retention-related morbidities. RESULTS: There was a significant increase in CO(2) elimination in the first 30 minutes of insufflation followed by a plateau for the remainder of procedure. There was no difference in CO(2) elimination in either procedure at any time interval. Patients with subcutaneous emphysema showed significantly greater CO(2) elimination, which decreased with desufflation. CONCLUSION: CO(2) absorption during laparoscopy did not depend on the route of surgery. Subcutaneous emphysema was strongly and independently associated with a greater degree of CO(2) absorption during laparoscopic surgery.

Evaluation of Antiasthmatic Activity of Curculigo orchioides Gaertn. Rhizomes.

The ethanol extract of Curculigo orchioides was evaluated for antiasthmatic activity by using various in vitro and in vivo animal models. In vitro models like isolated goat tracheal chain preparation and isolated guinea pig ileum preparation were studied to know basic mechanism by which extract shows relaxant activity. The study showed that extract is effective against histamine-induced contraction. In isolated goat tracheal chain preparation and isolated guinea pig ileum preparation extract exhibits maximum relaxant effect (p< 0.01) against histamine at concentrations 100mug/ml and 25mug/ml respectively. Animal studies involved use of histamine induced bronchoconstriction in guinea pigs, egg albumin induced passive paw anaphylaxis in rats and haloperidol-induced catalepsy in mice. These studies showed significant (p< 0.01) protection at lower doses while further increase in the dose level showed reduced activity. Biochemical estimations in milk-induced total leukocytes count and milk-induced differential leukocyte count were also studied. In this study there was maximum increase in leucocytes and lymphocytes (99%) and maximum decrease in eosinophils up to 0% at dose 375mg/kg p.o. body weight was observed. The results of these studies indicated usefulness of ethanol extract of Curculigo orchioides in asthma.

A study of blood glucose in paediatric laparoscopy.

BACKGROUND: There are few studies on stress responses to laparoscopic surgery in children. This study was conducted to assess the blood glucose levels in children undergoing laparoscopy. We also studied the effect of two different intravenous (i.v.) solutions on blood glucose in open and laparoscopic procedures. METHODS: One hundred and twenty healthy children, aged 2-12 years, undergoing either open or laparoscopic surgery, were randomized to receive either dextrose normal saline (DS) or Ringer's lactate peri-operatively (RL). All patients had blood glucose measurements performed immediately after induction but prior to the i.v. infusion of any fluid. Blood glucose was again measured 1 h after induction in the open cases and 1 h after insufflation in the laparoscopy cases. RESULTS: In the groups, baseline blood glucose values were comparable. In all groups, blood glucose concentrations increased from the immediate post-induction (baseline) values. When RL was infused, the 1-h blood glucose was higher in the laparoscopy group as compared with the open group. However, when DS was infused the difference between the 1-h blood glucose in the open and laparoscopic procedures was not statistically significant. In the laparoscopy group, the 1-h blood glucose value was significantly higher in the patients receiving dextrose solution. CONCLUSION: Laparoscopic procedures in children are associated with a rise in blood glucose levels similar to open surgery. The hyperglycaemic response was more pronounced when dextrose-containing solutions were infused peri-operatively.

Red cell hemolysis during processing and storage.

INTRODUCTION: Apart from the visual assessment, measurement of plasma hemoglobin in the supernatant from red cell units provides an objective measure of the extent of hemolysis during storage. STUDY DESIGN AND METHODS: Packed red cells (N=50), 25 units each in triple (CPD-A1 and SAGM) and quadruple (CPD-A1 and ADSOL) blood bags were evaluated for plasma hemoglobin by the tetramethylbenzidiene (TMB) method on day 1, 7, 14, 21 and 28 of collection. The hemoglobin, hematocrit, MCV, LDH and potassium levels were also noted. Whole blood units (N=25) were used as controls. RESULTS: Hemolysis increased in all the stored red cell units. Plasma hemoglobin increased significantly in the first week of storage. The hemolysis, LDH and potassium levels were found to be significantly higher in the red cell units harvested from the triple blood bags. However, on day 28 of storage, free hemoglobin in all the red cell units was much below the 0.8% hemolysis. CONCLUSION: Hemolysis of the red cells increases due to processing and during storage and is maximum during the first week. Adequate process control and proper storage facilities should be ensured to minimize the hemolysis of red cells during processing and storage.

Constitutional and somatic RB1 mutation spectrum in nonfamilial unilateral and bilateral retinoblastoma in India.

An epidemiologic survey has indicated a comparatively high prevalence of retinoblastoma (Rb) in Asian countries. Recently, the development of preventive strategies in nonfamilial Rb has become a major goal. The present studies were designed for identification and characterization of constitutional and somatic RB1 gene mutations by conventional cytogenetics, fluorescent in situ hybridization (FISH) and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP)-DNA sequencing. Of 34 patients 32 were nonfamilial and 2 were familial Rb. Maternal inheritance of del (13q14) was common. FISH was sensitive in detecting monoallelic RB1 deletion/deletion mosaicism as a first genetic hit in 20% of cases. Somatic and germline RB1 point mutations affected exons 3, 17, 20, and 21 and these were identified as novel mutations. Involvement of exon 20 as a predisposing mutation in sporadic unilateral retinoblastoma (URB) probably suggests the susceptibility of exon 20 to unknown etiologic factors in our population. A de novo RB1 deletion along with transmitted RB1 point mutation from an asymptomatic parent was identified as a unique predisposing RB1 mutation chimerism in a URB case that later evolved to bilateral retinoblastoma (BRB). The predisposing mutations such as del (13q), RB1 mono-allelic deletion and RB1 point mutation in sporadic Rb were de novo as well as transmitted mutations from asymptomatic/symptomatic parents. The RB1 mutation incidence was comparatively higher (25%) in nonfamilial Rb with emphasis on high prevalence in sporadic URB (18% versus 0%-9% in the literature series). The present studies demonstrated the efficacy of a multitechnique approach to detect various types of constitutional RB1 mutations such as RB1 deletion, deletion mosaicism, point mutation, mutation chimerism in patients of symptomatic/asymptomatic parents.

Imatinib mesylate in chronic myeloid leukemia: a prospective, single arm, non-randomized study.

Chronic myeloid leukemia (CML) is a hematopoietic stem cell disorder characterized by the balanced reciprocal translocation t (9:22). The resulting fusion gene, the BCR-ABL, is responsible for oncogenesis. Imatinib mesylate is a novel molecule, which inhibits the protein product of this fusion gene and hence has been used in the treatment of CML. The present study evaluates 174 patients with CML treated with imatinib mesylate. Of these 174 patients, 97 were in chronic phase, 47 in accelerated phase and 30 patients had blast crisis. Patients in chronic phase received imatinib mesylate in the dose of 400-mg daily, while those in accelerated phase and blast crisis received 600 to 800 mg daily. Of the 97 patients with chronic phase, 49 patients (50.5%) achieved a major (major + complete) cytogenetic response. Of the 47 patients in accelerated phase, 10 patients (21.3%) achieved a major cytogenetic response and in 30 patients with blast crisis, 7 (23.3%) achieved a major cytogenetic response. Dermatitis, mucositis, neutropenia and thrombocytopenia were some of the major toxicities. Of interest, 121 of the 174 patients (69.5%) developed generalized hypopigmentation. We conclude that imatinib mesylate is a safe and effective first-line therapy for chronic myeloid leukemia.

Comparative study of knowledge about different Child Survival and Safe Motherhood intervention in two groups of mothers.

A cross-sectional study was conducted in an urban field practice slum area served by Urban Health Centre (UHC) attached to the Dept. of Preventive and Social Medicine, T. N. Medical College and Nair Hospital, Mumbai, to compare the knowledge about different Child Survival and Safe Motherhood interventions in two groups of mothers. 152 mother who regularly attended antenatal check-up in UHC constituted study group and 153 mothers selected by individual matching constituted the control group. Significant differences in the knowledge of study and control groups of mothers were observed about some interventions like time of initiation of breast feeding, duration of exclusive breast feeding, age of starting weaning and number of OPV and DPT doses to be given till 1 year of age.

A study of mast cells in granulomatous lesions of skin, with special emphasis on leprosy.

76 skin biopsies that included material from 7 controls, 65 granulomatous skin lesions and 2 each of granulation tissue and chronic non-specific inflammation, were subjected to histopathological evaluation on haematoxylin and eosin and pertinent special stains. Mast cell study was done on slides stained by toluidine blue method, with special reference to their location, and morphology and cell count were done with the help of occculomicrometre. In normal skin, mast cell density was 11.43/mm2 with a range of 6-22/mm2 and an S.D. of 5.94. Highest value in the whole series was seen in TVC (66/mm2), followed by lupus vulgaris (50/mm2). Mast cell counts were normal in indeterminate and TT leprosy and showed a rise over the immunological spectrum BT to LL, with values in LL being 32.86/mm2 (28-40/mm2).