Cyto-histopathological correlation in palpable breast lesions

Background: Breast lesions are one of the most commonly encountered lesions in women which require prompt pathological confirmation by fine needle aspiration cytology (FNAC) and histopathological examination.Methods: We conducted a prospective study from January 2015 to December 2015. A total 98 cases included presenting with palpable breast lump in which 80 cases were also subjected to surgical biopsy or mastectomy.Results: Out of 98 cases, 34.7% benign cases, 59.2% malignant cases, and 6.1% non-neoplastic case were diagnosed cytologically in which 7 (7.1%) cases of mastitis, 2 cases (2%) of granulomatous mastitis, 22 cases (22.4%) of fibroadenoma, 11 cases (11.2%) of benign breast disease or fibrocystic disease, 10 (10.2%) cases of dyskaryotic changes , 45 cases (45.9%) carcinoma. Mean age was 46.4±14.2 years. Majority of cases 29(29.6%) belonged to 41-50 years age group. Majority of the masses were situated in the left breast (57.2%) in the upper outer quadrant (40.8%). In addition to breast lump, pain in 22 cases, bloody discharge in 5 cases, ulceration in 8 cases and nipple retraction in 11 cases were present. Histology was available for 80 cases in which 5 (6.3%) cases of non-neoplastic, 27 cases (33.7%) benign and 48 cases (60%) of malignant histology. FNAC proved to be 91.25 % sensitivity in the diagnosis of all breast lesions in our study.Conclusions: So we concluded that breast lesions are easily accessible to FNAC, which is an easy, cost effective and less time-consuming procedure. FNAC is used to diagnose both benign and malignant lesions.

Immobilization and stability studies of a lipase from thermophilic Bacillus sp: The effect of process parameters on immobilization of enzyme

A thermostable lipase was partially purified from the culture supernatant of a thermophilic Bacillus sp. The enzyme is optimally active at 60\ub0C and pH 8.0. The enzyme showed enhancement in activity in presence of benzene or hexane (30% v/v each). The activity (assayed by determining the release of pNP from pNP laurate) was stimulated up to 60% of these solvents in enzyme reaction mixture. The catalytic properties of this thermostable enzyme can be further improved via the use of different immobilization techniques and reaction conditions. Enzyme was immobilized on different solid supports and their enzyme activity and stability was compared. The enzyme was adsorbed on silica and HP-20 beads followed by cross-linking with gluteraldehyde on HP-20, which improved the thermostability of enzyme. The optimum pH (pH 8.5) was nearly same for aqueous and immobilized enzyme while optimum temperature was nearly 5\ub0C higher in case of immobilized enzyme. The immobilized/cross linked enzyme was more thermostable at 70 and 80\ub0C in comparison to aqueous and surface adsorbed lipase on silica and HP-20. The optimum temperature for esterification reactions was determined to be 60- 65\ub0C . Half-life of immobilized lipase was nearly 2.5 x higher than the aqueous enzyme at 70\ub0C . Esterification of methanol and oleic acid to methyl oleate by immobilized enzyme was studied in detail

Effect of Altitudinal and Seasonal Factors on Diatomological Mapping of Waterbodies: Implication in Drowning Investigations

Diatoms are an essential part of forensic investigation in drowning cases. They are used to differentiate between ante-mortem and post-mortem drowning and to ascertain the site of drowning. However, to do so it is imperative to perform the diatomological mapping of water sources in different regions. The issue of localizing a drowning site may be resolved with the help of diatomological monitoring and mapping of the water sources, which generates substantial references for meeting the ‘criterion of concordance’. The present study has generated a database of diatom diversity in relation to different seasons and altitudes. Water sampling was made from low, mid and high altitudes during summer and autumn seasons. Important information was found after a microscopic examination of diatoms in water samples from different water bodies at different geographical altitudes. Twenty-seven diatom genera have been observed in the water samples from the selected localities. The findings of the current study have exposed the significant effect of seasonal changes and varied altitudes on occurrence of diatoms. A variety of commonly occurring site restricted diatoms were identified in the water samples. This useful information can be significant while investigating drowning cases from this particular region when the drowning site is either questioned or unknown

FTIR Analysis of Some Pills of Forensic Interest

ABSTRACT This paper deals with the use of modern Fourier Transform Infrared (FTIR) spectroscopy with quick and easy Diamond Attenuated Total Reflectance (ATR) technique for generating a spectral library of some benzodiazepines of forensic interest. These types of drugs can be seized as bulk street drugs as well as traces found at the scene of crime. There can be legal queries regarding identification of these drugs which a forensic expert has to answer. In these cases, the standard reference database is required for comparison. The modern FTIR systems with diamond ATR proves to be a rapid, sensitive and non-destructive analysis of samples with very little effort. This spectral library can be used as a reference library when an unknown sample is suspected of being benzodiazepine

DNA breaks regulate neuronal activity: NMDA-receptor and ionizing radiation-mediated DNA double-strand breaks govern activity regulating early-response gene expression

DNA (Deoxyribonucleic acid) double-strand breaks (DSBs) are the hazardous form of DNA damage endured on multiple occasions by both proliferating and post-mitotic cells. Unlike replicating cells, which can employ sister chromatids as a repair template, post-mitotic cells such as adult neurons could suffer lethal effects from error-prone repair of non-homologous end-joining (NHEJ) machinery. Hence, until recently, DSBs were considered only the results of pathological activity in post-mitotic cells. However, recent shreds of evidence also show the presence of neuronal activity-induced and type 2 topoisomerase beta (Top2β) generated transient physiological DSBs, which regulate the expression of multi-functional early response or immediate early genes (IEGs). Till yet, these DSBs driven gene regulations have only been studied in cortical cultures. Among many, one of the functions of effectors IEGs is the regulation of neuronal activity by activation of secondary response genes. Therefore, hypothesizing that neuronal activity induced DSB promotes certain IEGs expression and further these genes could regulate the activity, we aimed to, i) first validate the correlation of neuronal activity generated DSBs with IEGs expression in neuronal model systems other than cortical cultures ii) investigate the effects of altering DNA DSB's status on neuronal network activity. As a model system for our analysis, we employed 14 days-in-vitro (DIV) mice hippocampal cultures (HCs), where the expression of IEGs has been reported before. As only specific neuronal activity could lead to induction of physiological DSBs, we primarily focused on the DNA breaks generated by N-Methyl-D-aspartate receptors (NMDARs) dependent cAMP (Cyclic adenosine monophosphate) response element-binding protein (CREB) activity. For our investigation, we employed a combination of Microelectrode arrays recordings (MEA) for neuronal network activity measurements, Gamma-H2AX staining to monitor DSBs' presence and messenger Ribonucleic acid (mRNA) and protein analysis for IEGs expression. Following NMDAR activation, we detected an increase in DSBs status, which correlated with the expression of IEGs- c-Fos, Egr1 and c-Jun and upregulation in neuronal activity. Additionally, the DSBs status and IEGs expression were also analyzed following the inhibition of essential synaptic plasticity-associated components, CREB and the GluN2B subunit of the NMDAR. Our results showed that the deficiency in any of these factors leads to a decrease in certain IEGs expression, likely by causing changes in DSBs status. Further, to investigate if DSBs status alteration can affect neuronal firing activity by perturbating IEGs expression, we incubated HCs with either etoposide, a chemotherapeutic agent that generates DSBs on the site of Top2β activity, Top2β (small interfering RNA) siRNA and NHEJ inhibitor (NU7441). Our data discovered that a perturbation in either the induction and repair of the DSBs following NMDAR activation contributes to both altered IEGs expression and neuronal activity. Furthermore, the IEGs expression was also monitored in the Severe combined immunodeficiency (SCID) mice model. Our novel results revealed that these mice models have a low level of IEGs expression, most likely due to the inherent deficiency in the components of NHEJ. This could lead to severe learning and memory problems that these mice possess. Thus, our study provides experimental evidence on the role of synaptic NMDARs mediated DSBs in mediating IEGs expression and further how a perturbation in the DSBs induction and repair could contribute to an altered pattern of neuronal activity in HCs. Furthermore, by assessing these connective links of receptor activity with gene expression and neuronal recordings, our study validates the multi-purpose use of such an in-vitro model to elucidate changes in cellular and electrophysiological levels. In the second part of this work, we investigated the effect of low-dose radiation exposure on IEGs expression. Ionizing radiation (IR) is an effective treatment for various tumors infested in the central nervous system (CNS). Proceeding the clinical trials, multiple literature pieces have shown how even low dose exposure could alter synaptic plasticity, a basis for cognitive functioning. However, it becomes complicated to assess the connective link between irradiation and synaptic plasticity-associated components due to IR's multi targets. Therefore, to understand how irradiation can have a lethal effect on cognitive domains, we focussed on the expression of the genes that are easily activated by various stimuli and play a role in learning and memory acquisition. For this, we studied the effect of low to moderate IR doses on the expression of IEGs in 14 DIV HCs. Results of immunostaining against Gamma-H2AX following radiation exposure revealed that a single 2 Gy dose generates many DSBs in the post-mitotic neuron post-60-minute (min) exposure. However, the expression of IEGs such as c-Fos and Egr1 as assessed by mRNA and protein analysis following the same dose radiation exposure is downregulated. As both of these genes are the regulator of neuronal activity, we next analyzed the neuronal network activity following irradiation. Our data revealed that a single dose of 2 Gy is sufficient to cause a decrease in neuronal network activity after 60 min of exposure. NMDAR plays a vital role in IEGs expression; therefore, we investigated if NMDA incubation can counteract the radiation alleviated IEGs expression level. Consequently, we further investigated the simultaneous impact of NMDAR activation and irradiation on the control of IEGs expression by DSB's and mRNA analysis. Our results revealed that 60 min of continuous NMDA treatment counteracts radiation-induced alleviation of IEGs expression. These findings are the first to reveal changes in IEGs expression and neuronal activity in HCs following 1-hour post- single-irradiation events. Further, it hints towards the interaction radiation has with the NMDAR pathway for the gene expression changes