Diagnostic ability of real-time quantitative polymerase chain reaction versus immunohistochemistry for Ki-67 assessment in breast cancer: An Indian perspective

Background & objectives: Breast cancer is the most common cancer of women. Inferior prognosis in some patients has been attributed to the higher proliferative capability of the tumour. Immunohistochemistry (IHC) for Ki-67, despite being a simple and cost-effective method, has not become a valid tool to evaluate this biomarker. This is ascribed to variation in pre-analytical and analytical techniques, variable expression, hotspot distribution and inter-and intra-observer inconsistency. This study was aimed at defining the analytical and clinical validity of real-time quantitative polymerase chain reaction (RT-qPCR) as an alternative to IHC evaluation. Methods: This study included a total of 109 patients with invasive breast cancers. Ki-67 IHC visual assessment was compared with the mRNA value determined by RT-qPCR. Concordance between both the methods was assessed. Receiver operating characteristic (ROC) curve analysis and Cohen's kappa value with intraclass correlation were performed. Results: The threshold value for Ki-67 by RT-qPCR obtained by ROC curve was 22.23 per cent, which was used to divide breast cancer cases into high proliferative and low proliferative groups. A significant correlation was observed between both the breast cancer groups formed using RT-qPCR threshold as well as median laboratory value of Ki-67 labelling index by IHC. Interpretation & conclusions: The study results showed a significant correlation between the two methods. While IHC is subject to technical and interpretative variability, RT-qPCR may offer a more objective alternative.

Modifiable risk factors of noncommunicable diseases in urban adolescents of Girdhar Nagar, Ahmedabad: A cross-sectional study

Background: Noncommunicable diseases (NCDs) have become a major health burden globally. The four main NCDs (heart disease, stroke, diabetes, and cancers and chronic respiratory diseases) share four modifiable risk factors: Tobacco use, harmful alcohol use, physical inactivity, and unhealthy diet. To minimize the burden of NCD epidemic, main risk behaviors common among young people must be curbed. Measurement of NCD risk factors is necessary to plan out the interventions and would also serve as a stepping stone in achieving primordial prevention of NCDs. Objective: The objective of the study was to measure the modifiable risk factors of NCDs among school-going adolescents of urban Ahmedabad. Materials and Methods: This cross-sectional study was conducted from November 2016 to January 2017 among two schools of Girdhar Nagar ward, Ahmedabad. 581 adolescents of a private and a government-run school, studying in class 8th–10th, of ages 12–16 were interviewed using GSBHS tool after taking informed consent. Appropriate anthropometric and blood pressure (BP) measurements were taken. Statistical analysis: Epi info ver. 7.0 and MS Excel 2007 used for analysis. Chi-square test, t-test, and multiple regression were applied. P < 5% was considered significant. Results: 52% (303) of the subject studied were boys and 48% (278) were girls. Among all, 66% (384) were physically inactive, 88% (511) were not consuming ≥5 servings of fruits and vegetables/day, 65% (376) and 78% (452) regularly consumed soft-drinks and fast food, respectively. 1.5% (9) of the subjects smoked and 3%(15) consumed smokeless form of tobacco. No student admitted consuming alcohol. 11% (65) were overweight and 13% (73) were obese. Raised systolic BP (>140 mmHg) was found in 3% (18) and raised diastolic BP (>90 mmHg) was found in 0.5% (3) of study subjects. Conclusion: One or more risk factors were found to be prevalent in all subjects. The presence of one or more modifiable risk factors in every adolescent point toward the dire need of school-based interventions for the prevention of NCDs in future adults.

Isolated tubercular splenic abscess: Can we defer splenectomy? Our single experience with anti-tuberculous therapy alone.

Isolated splenic tuberculosis is an exceedingly rare clinical condition. Microbiological confirmation of diagnosis in such cases is quite difficult. We encountered the case of a 35-year-old female, who presented with persistent low-grade fever and weight loss. The CT scan of the abdomen revealed multiple hypodense splenic lesions. No primary focus of infection was detected in any other organs. Fine needle aspiration of splenic lesion revealed acid-fast bacilli on Ziehl-Neelsen stain. With anti-tuberculous therapy, the lesions regressed significantly in size. We stress that splenic tuberculosis should be considered as a diagnostic possibility even in immunocompetent individuals and choose combination antituberculous therapy as the first line treatment with consideration of splenectomy depending on response.

Development and characterization of two new cell lines from common carp, Cyprinus carpio (Linn)

Two new cell lines (CCF and CCH) were established from fin and heart tissues of common carp, Cyprinus carpio. The cells were optimally maintained in Leibovitz-15 medium supplemented with 10 percent fetal bovine serum (FBS) and 10 ng/ml of basic fibroblastic growth factor (bFGF). The effects of temperature, concentration of FBS and bFGF on the growth of CCF and CCH cells were examined. The temperature ranged from 24 to 32 °C for good growth of the cells. The growth rate of cells was higher in medium containing 10 percent FBS and the addition of bFGF to the medium significantly increased the growth rate. The CCF cells were found to be epithelial, while the CCH cells were fibroblastic in nature. The cytogenetic analysis of the cell lines revealed a diploid number of 100 chromosomes in C. carpio. The viability of CCF and CCH cell lines were 70 and 72 percent, respectively, after six months of storage in liquid nitrogen (-196 ° C). Molecular characterization of the cell lines using 16S rRNA and Cytochrome Oxidase Subunit I (COI) revealed the origin of the cell lines. These new cell lines will be useful for isolation of fish viruses and other in vitro biotechnological studies.

Relation between insulin secretory capacity and microangiopathy in young diabetic patients in Bangladesh.

The pathogenesis of diabetic retinopathy is still unclear. The relative role of duration of diabetes, glycemic and insulinemic status in the etiopathogenesis of retinopathy is to be clearly understood. Hypertension, hyperlipidemia, pregnancy and age at diagnosis have been thought to be factors associated with diabetic retinopathy. Taking advantage of the availability of a group of young lean, normotensive and generally normolipidemic subjects in Bangladeshi population, the present study was undertaken to investigate the relationship between insulin secretory capacity and microvascular damage in the pathogenesis of diabetic retinopathy. A total number of 91 diabetic subjects, diabetes diagnosed before the age of 30 years, were recruited form the out-patient Department, BIRDEM Hospital. Diabetic subjects recruited were new- to previously diagnosed cases with duration of diabetes around 8 years. Insulin secretory capacity was assessed by C-peptide and C-peptide was estimated by ELISA method. Urinary albumin was measured by Immunoturbidetric methods. The subjects were grouped on the basis of duration of diabetes and presence (DR) and absence (NDR) of diabetic retinopathy. Subjects were also divided on the basis of albumin-creatinine ratio (ACR) level. ACR level was observed sharply increased with the duration of diabetes and showed a peak after 8 years. In the ACR positive cases 95.25% of diabetic patient had retinopathy. Both retinopathic and ACR positive subjects had low insulin secretory capacity as evident by absolute C-peptide level and C-peptide glucose ratio. Retinopathy was found increased with generalized vasculopathy. Both microvasculopathy and retinopathy seemed to occur at around 8 years. Insulin secretory capacity in particular seemed to have predominant determinant effect in the etiopathogenesis of diabetic retinopathy.

Maternal knowledge on risk behavioural practices and it's association with diarrhoea in a rural community of West Bengal, India.

Knowledge of rural mothers related to five diarrhoeagenic risk behaviours, identified in an earlier study, was ascertained. A high proportion of mothers (67%-79%) had knowledge about risk of bottle feeding, non-use of soap for cleaning feeding containers, storage of drinking water in wide-mouthed vessels and indiscriminate disposal of children's faeces. However, only around 31% of mothers were aware about danger of using pond water for cleaning feeding containers. Risk behavioural practices were less amongst mothers who had knowledge about them. Risk of diarrhoea amongst children of mothers having risk practice without knowledge as compared to those who utilised their knowledge to avoid risk practice was found significantly higher (p < or = 0.005) except for bottle feeding (p = 0.330). The results of this study indicate that children can be protected significantly from diarrhoea if mothers' diarrhoeagenic behaviours can be altered through educational intervention.

Effects of some psychotropic agents on cognitive functions in rats.

The effects of some psychotropic agents were evaluated on cognitive function in rats. Acute administration of antipsychotic agents chlorpromazine, haloperidol or pimozide-all inhibited the active avoidance response as elicited by pole climbing test-the least effect being seen with pimozide. The antianxiety benzodiazepines, lorazepam and alprazolam, also attenuated active avoidance learning with the latter showing lesser effects. Maze task performance was differentially affected by these psychotropic drugs with maze traversing time being increased significantly after all drug treatment, except alprazolam.

Antinociceptive action of Azadirachta indica (neem) in mice: possible mechanisms involved.

Azadirachta indica (AI, Neem) was tested for analgesic potency in experimental pain models in mice. In the glacial acetic acid (GAA) induced writhing test, AI (10, 30 and 100 mg/kg) dose-dependently reduced both the incidence and the number of writhes. Similarly, AI, at the dose levels tested, also enhanced tail withdrawal latencies in the tail-flick test for nociception. In the interaction studies, pretreatment with the opioid antagonist, naloxone (1 mg/kg) and the central noradrenaline depleter, DSP-4 (50 mg/kg) attenuated AI analgesia by differential degrees in both experimental models, whereas, the serotonin synthesis inhibitor, PCPA (300 mg/kg) potentiated the same. These results suggest that both central and peripheral mechanisms and complex neural pathways, opioid and non-opioid, may be involved in AI induced analgesia.

Fat and oil inhibit phosphine release from aluminium phosphide--its clinical implication.

Poisoning by suicidal or accidental ingestion of aluminium phosphide (AlP) is a frequent medical emergency seen all over the world. AlP, a grain fumigant and rodenticide, on exposure to moisture, liberates highly toxic gas, phosphine. The rapidly inhibits mitochondrial respiration and has cytotoxic action. No specific antidote is known against it till date and prognosis depends much on dose and time lag between AlP ingestion and the stomach wash in the hospital (critical period). Physicochemical properties of AlP and nonmiscibility of fat and water promoted us to study the effect of different fats and oils as possible antidotes to inhibit phosphine liberation. In vitro experiments revealed that vegetable oils and liquid paraffin were much more effective than butter and ghee in inhibiting release of phosphine from AlP. These findings may have significant clinical implication.