Clinico-pathological and Immunophenotype Analysis of Hairy Cell Leukemia Cases – A Single Institute Experience

Background: Hairy cell leukemia is a mature lymphoid B cell disorder, characterized by hairy cells, a specific genetic profile, different clinical course and the need for an appropriate treatment. It is seen primarily in elderly, characterized by a triad of splenomegaly, pancytopenia and monocytopenia. Aim: To evaluate clinicopathologically and with immunophenotyping hairy cell leukemia cases received at our institute in conjunction with similar studies. Material and methods: This is a retrospective study which included 7 cases over a period of 3 years (2019-2021) confirmed on morphology and flow cytometry. Results: The study revealed 7 cases which showed patients with age ranging from 34-65 years. M:F ratio was 6:1. Two cases were covid positive (28.5%). Most of the cases presented with fever, weakness (28.5%). Splenomegaly was seen in three of the cases (42.6%). Laboratory investigations revealed anemia in 71% cases, leucopenia in 56.8%, lymphocytic prominence in 100% and pancytopenia in 14.2%. One patient presented with leukocystosis (14.2%). Marrow was hemodiluted and aparticulate in 3(42.6%) cases. Hairy cells were seen on morphology of peripheral smear and marrow aspirate. On flow cytometry, CD5 negative in all cases (100%), CD10 positive in 2(28.5%) and CD23 in 2 cases (28.5%). Few cases confirmed BRAF v600e mutations. Conclusion: Unusual findings like leukocytosis, absence of spleen, presence of lymphadenopathy can be present in hairy cell leukemia. Classical fried egg appearance in trephine biopsy may not be afeature in all the cases. CD123 is expressed in covid patients unlike other studies and further research is needed to establish the loss of CD123 in covid patients.

Clinicoepidemiology, clinical profile and outcome of venomous snake bite in children in Konaseema region of Andhra Pradesh, India

Background: Venomous snake bite is an acute medical emergency, and children are more quickly and severely affected than adult. Venomous snake bite is responsible for 28.5 deaths per 1000 snake bite death among 5-15years old children. Most of them used to be from rural India and is more common in school going children. But there is no study regarding clinic epidemiology and treatment outcome among pediatric age group in this part of Andhra Pradesh. This retrospective descriptive has been conducted to evaluate the clinic epidemiology, clinical presentation, complication and treatment outcome in snake bite in pediatric patients in our set up.Methods: Present study is a retrospective, descriptive study conducted in the department of paediatrics Konaseema institute of medical science Amalapuram Andhra Pradesh, between 2006 to September 2018. This study protocol is approved by the institutional ethics committee. All the clinical parameters like, age, sex, season of bite, time of bite, socio economic statue, region, site, common clinical presentation, complications, reaction to ASV outcome of treatment were obtained from patient's case record and was analysed.Results: In present study total 166 snake bite patient case records were evaluated, out of this 166 snake bite cases, 28 bites were by unknown snakes. Demographic profile of patient shows that out of 70 cases, 49 cases were in male child and 21 cases were in female child. Regarding clinical profile of the children with snake bite as mentioned in Table 3, local pain was present in (n=70) 100 % patients. Local blending was presenting symptom in 64.28 %(n=45) patients.Conclusions: From present study authors conclude that snake bite in children is more common among school going children, during monsoon and post monsoon, and in rural area. In our region vasculotoxic snake envanomation is more common and presented with cellulitis which required surgical intervention in most of circumstances. In neurotoxic envenomation some patient developed respiratory paralysis which required ventilatory support.

Prospective study of spectrum, clinico-epidemiology, profile complication and outcome of pesticide poisoning in children

Background: Konaseema region of Andhra Pradesh is a rural area having well irrigated lands and rice fields. Agriculture is the major some of income, and pesticide use is high.' As per one report of deccan chronicle (a daily newspaper) Andhra Pradesh and Telangana accounts for 24% share of pesticide consumption in the country. Pediatric pesticide poisoning is under reported in India as well. In this background resent study has been designed to study clinic-epidemiology, profile, complication and treatment outcome of pesticide poisoning in Konaseema region of Andhra Pradesh.Methods: In present study clinico-epidemiology, clinical profile, complication and outcome of the patient admitted in the Department of Emergency medicine and Paediatric were evaluated over a period of 12years.Results: Most of the children were above 5years of age that is 74.0% (n=40), rest were below 5years of age. Male children out numbered female children and accounted for 77.78% (n=42). Accidental poisoning was more than suicidal poisoning that is 94.59% of the patients. 9.35% patient (n=5) developed respiratory failure and required ventilator support. Non-carcinogenic pulmonary edema was present in 8 (14.81%) patients. Four patients have atropine toxicity, electrolyte in balance was found in 7.4% (n=4) patients. Acute kidney injury was present in two patients, 5.5% (n=3) patients developed cardiac arrhythmia.Conclusions: Chlorpyrifos was common agent which is responsible for poisoning most common complication in our patients were respiratory complications which required ventilatory support. Lack of information, improper disposal of container, non-existence of training and regulation is supported to the cause of accidental exposure of pesticide to paediatric patients.

Relationship between promoter methylation & tissue expression of MGMT gene in ovarian cancer.

Background & objectives: Epigenetic alterations, in addition to multiple gene abnormalities, are involved in the genesis and progression of human cancers. Aberrant methylation of CpG islands within promoter regions is associated with transcriptional inactivation of various tumour suppressor genes. O6-methyguanine-DNA methyltransferase (MGMT) is a DNA repair gene that removes mutagenic and cytotoxic adducts from the O6-position of guanine induced by alkylating agents. MGMT promoter hypermethylation and reduced expression has been found in some primary human carcinomas. We studied DNA methylation of CpG islands of the MGMT gene and its relation with MGMT protein expression in human epithelial ovarian carcinoma. Methods: A total of 88 epithelial ovarian cancer (EOC) tissue samples, 14 low malignant potential (LMP) tumours and 20 benign ovarian tissue samples were analysed for MGMT promoter methylation by nested methylation-specific polymerase chain reaction (MSP) after bisulphite modification of DNA. A subset of 64 EOC samples, 10 LMP and benign tumours and five normal ovarian tissue samples were analysed for protein expression by immunohistochemistry. Results: The methylation frequencies of the MGMT gene promoter were found to be 29.5, 28.6 and 20 per cent for EOC samples, LMP tumours and benign cases, respectively. Positive protein expression was observed in 93.8 per cent of EOC and 100 per cent in LMP, benign tumours and normal ovarian tissue samples. Promoter hypermethylation with loss of protein expression was seen only in one case of EOC. Interpretation & conclusions: Our results suggest that MGMT promoter hypermethylation does not always reflect gene expression.

Association of vascular endothelial growth factor single nucleotide polymorphisms on the prognosis of breast cancer patients.

BACKGROUND: Vascular endothelial growth factor (VEGF), a major mediator of vascular permeability and angiogenesis, may play a pivotal role in mediating the development and progression of breast cancer. In the present study, we examined the genetic variations of the VEGF gene to assess its possible relation to breast cancer. MATERIALS AND METHODS: A total of 200 patients with histologically confirmed cases of breast cancer and 200 healthy women were genotyped for VEGF single nucleotide polymorphisms (405G > C and −1154G > A) by polymerase chain reaction‑restriction fragment length polymorphism analysis. Pre‑operative plasma VEGF levels were determined by enzyme‑linked immunosorbent assay in 200 women with breast cancer and in 200 normal female controls. RESULTS: The genotype frequencies of the +405G > C, −1154G > A polymorphisms did not show a significant deviation from the Hardy‑Weinberg expectation. The minor allele frequencies of the +405G > C and −1154G > A polymorphisms among cases and controls were 33.5% (C allele), 31.5% (A allele) and 35% (C allele), 34.5% (A allele) respectively. +405GG and −1154GG genotypes were associated with higher levels of VEGF among breast cancer cases and controls. Increased plasma VEGF levels were significantly associated with, clinical stage of the disease (P = 0.035). CONCLUSION: Although none of the polymorphisms were significantly associated with breast cancer, some of the VEGF genotypes may influence tumor growth through an altered expression of VEGF and tumor angiogenesis.

Multidisciplinary approach in the long-term management of intrahepatic stones: Indian experience.

BACKGROUND: Intrahepatic stones, though common in East Asia, are uncommon in India. There is paucity of data from India regarding the treatment and long-term outcome of patients with intrahepatic stones. METHODS: We retrospectively analyzed medical records of 35 patients with intrahepatic stones who had been treated surgically. Endoscopic biliary drainage had been used in patients who presented with acute cholangitis. Intraoperative stone clearance was confirmed by choledochoscopy and intraoperative cholangiography. Outcome of surgery, frequency and subsequent management of recurrent intrahepatic stones, and factors associated with stone recurrence and cholangitis were analyzed. RESULTS: Twenty-one (60%) patients had bilobar disease. Eight patients underwent hepatectomy and 16 hepatico-jejunostomy with access loop; 12 of these were jejuno-duodenal anastomoses. Postoperative morbidity was observed in 10 patients (29%). Recurrence of stones occurred in 12 patients (34%) and cholangitis in 9 patients (26%). Presence of bilobar disease and associated biliary strictures were associated with recurrent cholangitis (p< 0.05). Two patients (6%) required re-operation for recurrent cholangitis. Complete removal of recurrent stones using conventional endoscope was possible through jejuno-duodenostomy in all 5 cases who had this type of access loop construction. CONCLUSION: Surgical treatment for intrahepatic stones depends on the site of involvement. Construction of a jejuno-duodenal access loop in patients with bilobar disease and intrahepatic strictures is helpful in facilitating postoperative stone clearance. A multidisciplinary approach is beneficial especially when the disease is bilobar and recurrent in type.

Active immunization against riboflavin carrier protein results in peri-implantation embryonic loss leading to pregnancy termination in rats: use of alternate adjuvants.

To investigate the mechanism of pregnancy termination following immuno-neutralization of riboflavin carrier protein (RCP) and to use acceptable adjuvants, we actively immunized female rats with reduced and carboxymethylated RCP (RCM-RCP) using various adjuvants (during primary immunization) such as sodium phthalylated lipopolysaccharide (SPLPS), purified S. typhi outer membrane proteins (porins) and a combination of them. Rats (5-14 per group) were immunized with alugel adsorbed RCM-RCP (100 microg/dose) either alone or with SPLPS or porins or SPLPS+porins. Control animals received RCM-RCP emulsified with Fruend's completelincomplete adjuvants (FCA/FIA). All animals received five boosters at intervals of 21 days. The lowest (4 X 10(-3)) and the highest (> 70 X 10(-3)) anti-RCM-RCP antibody titers were observed in alugel adsorbed-RCM-RCP group and control groups, respectively. Immunized animals showed reduced fertility following 3rd, 4th and 5th boosters. Reduction in fertility was 30-60% in alugel adsorbed RCM-RCP group, 90-100% in FCA-RCM-RCP group and 80-90% in SPLPS+porins group. Fertility reduction was not strictly correlatable with the serum antibody titers. RCP-specific IgG could be localized in the uterine endometrial glands and luminal epithelial cells in the immunized animals. Animals in the FCA/FIA group showed abnormal implantation/resorption sites and their histological sections showed degenerated embryos. But, day 5 preimplantation embryos were normal. These results show that (a) SPLPS+porins can be used as adjuvants in place of FCA/FIA for active immunization against RCM-RCP and (b) early termination of pregnancy in the immunized animals is due largely to the failure of normal embryo implantation.

Cloning and characterization of mycobacteriophage I3 promoters.

Restriction fragments of mycobacteriophage I3 DNA capable of initiating transcription have been cloned into a promoter selection vector of Escherichia coli, and selected on the basis of development of resistance to chloramphenicol. The growth pattern of these 'promoter clones' on a concentration gradient of chloramphenicol and the biochemical assays of the chloramphenicol acetyl transferase have permitted the assessment of their relative promoter strengths. DNA sequence analysis revealed significant homology of these promoters to the -35 regions of the mycobacterial--and E. coli promoter consensus, but less so the -10 region. Based on the sequence of phage I3 promoters identified here and the reported sequences of mycobacterial promoters, a promoter consensus for mycobacteria has been generated.

Hepatitis E virus: epidemiological, clinical and serological studies of north Indian epidemic.

BACKGROUND: A large waterborne epidemic of viral hepatitis occurred in the city of Karnal (Haryana) from February to April 1987. An attempt was made to study the epidemic clinically, serologically and etiologically. METHODS: A house-to-house search of the city was conducted for the detection of acute hepatitis cases. Patients willing to give blood samples for liver function tests were studied. RESULTS: A total of 1273 persons (0.79% of persons surveyed) were affected by viral hepatitis. Of the 477 clinically and biochemically documented cases, more than 75% were adults, while only 11% were less than 10 years old. Children below 15 years of age and females had a significantly higher incidence of anicteric hepatitis. Serological markers for acute hepatitis A and B viruses were absent in 85% of patients. Antibodies to hepatitis E virus (HEV) were detected in 84% of acute phase sera studied by indirect immunofluorescence assay. Liver biopsy showed characteristic features of cholangitic hepatitis. Three of 19 pregnant females who developed hepatitis died. No residual clinical or biochemical abnormality was detected in any of the patients followed up for 8 months. This epidemic correlated with the timing of unsupervised digging of lanes to provide new tap water connections to houses in the congested area of the city. This resulted in damage to the sewerage system and leakages which contaminated drinking water supply. CONCLUSION: HEV was transmitted by contaminated drinking water in this epidemic. Most affected individuals were adults, and recovered without sequelae. Affected pregnant women had a worse outcome.

Epidemiological study of parasitic infestations in lower socio-economic group in Chandigarh (north India).

When stool samples from 970 subjects belonging to lower socio-economic groups were examined for parasites, a total of 121 subjects (12.5%) i.e., 57 (12.1%) males and 64 (12.9%) females showed positive results. The overall prevalence of parasitic infestation did not correlate with sex, caste or religion and living conditions. However, the prevalence was higher in hospital employees residing in well sanitated area. Giardia lamblia (69.5%), Entamoeba, histolytica (15.7%), Hymenolepis nana (12.4%), Ancylostoma duodenale (10.7%), Ascaris lumbricoides (8.3%) and Taenia (0.8%) were the parasites seen. Mixed infections were seen in 9 subjects. Twenty families of the 196 studied had more than one family member positive for parasites. Asymptomatic positivity was high amongst all groups of subjects, and with all parasites.