PREPARATION AND CHARACTERIZATION OF ALGINATE CHITOSAN CROSSLINKED NANOPARTICLES BEARING DRUG FOR THE EFFECTIVE MANAGEMENT OF ULCERATIVE COLITIS

Objective: Delivery of anticancer molecule to the liver remains a “holy grail” in molecular medicine and nanobiotechnology with conventional therapy, as conventional cancer chemotherapy does not prove effective as drug molecule does not reach to the target site at therapeutic concentration. Tumor vasculature differs from the vasculature of normal tissue both in morphology and biochemistry. Most of these differences appear too related to angiogenesis (formation of new blood vessels from pre-existing ones). For the present study nanoparticles (NPs) were chosen as a delivery system, because they have many advantages, e. g. they can pass through the smallest capillary vessels because of their ultra-tiny volume, can penetrate cells and tissue gap to arrive at, pH, ion and/or temperature sensitivity of materials, can improve the utility of drugs and reduce toxic side effects. Methods: PLGA (poly lactide co glycolic acid) was used for the preparation of NPs because of its biodegradability and biocompatibility. It degrades by hydrolysis of ester linkages in the presence of water in to two monomers lactic acid and glycolic acid. There are a number of ligands available for hepatic delivery, among them lactobionic acid (containing galactose moiety) was selected for present work. Preparation of plain nanoparticles was carried out using emulsification–diffusion method. Optimization of the polymer concentration is the first step during the study and it was performed by varying the polymer concentration where as keeping other variables constant. The prepared formulation was optimized on the basis of particle size and polydispersity index. Amount of drug was optimized on the basis of particle size and percentage entrapment efficiency. Results: Particle size and zeta potential of the nanoparticle were determined by zetasizer showed that particles are in nano range (blow 200 nm) and have acceptable range of zeta potential. Shape and surface morphology were determined by TEM and SEM analysis. The conjugation of lactobionic acid with PLGA polymer was proved by FTIR. The in vitro release profiles of entrapped drug from formulations were determined using dialysis membrane. For stability studies, the LDNPs (conjugated NPs) are stored at the temperatures 4±1 °C and room temperature. Human hepatoma cell line HepG2 by SRB assay was selected and it clearly suggests a dose dependent cytotoxicity response i.e. decrease in cell survival fraction with increasing concentration of drug. The in vivo study are important in evaluating the targeting efficacy of designed dosage form and also helps in establishing the correlation between the results obtained from in vitro experimentation to that from in vivo studies. The formulations were administered by tail vein to mice of four groups Group I: PBS 7.4 (control); Group II: 5-FU solution; Group III: DNPs; Group IV: LDNPs. Conclusion: The proposed targeting strategy is expected to enhance the therapeutic index of conventional anticancer drug as well as reduce its cytotoxic effects to normal cells

Megakaryocyte in Peripheral Blood Smears – A Report of Two Cases

Myeloproliferative neoplasm (MPN), a clonal hematopoietic stem-cell disorder, results from the proliferation of one or more hematopoietic series of cells like erythroid, granulocytic or megakaryocytic series. Megakaryocytes (MGK) are large polypoidal cells seen within bone marrow aspirate (BMA) smears. We are presenting here two cases of MGK in peripheral blood smears (PBS), one with MPN and the other in a case of chronic myeloid leukemia (CML) with blast crisis. MGK in PBS is rare and is not always associated with neoplasm. It can be due to increased MGK differentiation due to reactive etiology

Flow Cytometer: The Need of Modern Hematology Laboratory

Indeed, laboratory diagnosis is rapidly changing from what it was in the past to the present. Nowadays, flow cytometer (FCM) has become a novel introduction to the modern diagnostic technique, mainly in the field of hematology. In case of acute leukemia, peripheral blood, bone marrow examination, cytochemistry and immunohistochemistry for differentiation of myeloid or lymphoid lineage is required, which is feasible by flow cytometry. It has dramatically improved the diagnostic efficiency and reduced the duration of sampling along with better diagnostic outcomes as well as provided efficient therapeutic monitoring of any drug or drug regimen. It has also opened some more sensitive therapeutic plans, like monitoring “Minimal residual disease (MRD)”, which is not possible without FCM. Detection of MRD has led to improved overall survival of patients. It has also opened up huge opportunities for research, which has become an important part of academic curriculum nowadays. Considering the importance and absolute necessity for better outcomes in hematology, the knowledge of basic principle of FCM becomes indispensable. Here, we try to elucidate the elementary components of this technique and also highlight its uses.&nbsp

Role of CRISPR Technology in Gene Editing of Emerging and Re-emerging Vector Borne Disease

Vector borne diseases are rampant across the world. Due to spread and establishment of vector species in different geographical areas, vector adaptation and resistance towards many insecticides the only option left is vector control for various vector borne diseases. Recent advancement in the field of genome editing have provided a variety of tools like, CRISPR, a novel genome editing techniques which can be applied for the control and prevention of many deadly diseases like dengue, chikungunya, filariasis, Japanese encephalitis and Zika. The present chapter is aimed to discuss the recent advancement in genome editing tools such as, their application, challenges, and limitations in vector control. Additionally, this chapter would potentially be advantageous to understand the hurdles, knowledge gaps in eliminating vector borne disease

Flow Cytometer: The Need of Modern Hematology Laboratory

Indeed, laboratory diagnosis is rapidly changing from what it was in the past to the present. Nowadays, flow cytometer (FCM) has become a novel introduction to the modern diagnostic technique, mainly in the field of hematology. In case of acute leukemia, peripheral blood, bone marrow examination, cytochemistry and immunohistochemistry for differentiation of myeloid or lymphoid lineage is required, which is feasible by flow cytometry. It has dramatically improved the diagnostic efficiency and reduced the duration of sampling along with better diagnostic outcomes as well as provided efficient therapeutic monitoring of any drug or drug regimen. It has also opened some more sensitive therapeutic plans, like monitoring “Minimal residual disease (MRD)”, which is not possible without FCM. Detection of MRD has led to improved overall survival of patients. It has also opened up huge opportunities for research, which has become an important part of academic curriculum nowadays. Considering the importance and absolute necessity for better outcomes in hematology, the knowledge of basic principle of FCM becomes indispensable. Here, we try to elucidate the elementary components of this technique and also highlight its uses

Reliability and reproducibility of intraocular pressure measurements using the Goldmann applanation tonometry in glaucoma suspects

Aims: This study aimed to examine the intra-observer reliability of three ophthalmologists who measured intraocular pressure (IOP) using the Goldmann applanation tonometry (GAT) and the inter-observer variability among them during three visits of glaucoma suspects. Methods: This prospective observational study evaluated the reliability and consistency of the IOP measurements made by three trained observers using the GAT. The three observers recorded IOP readings for 100 patients during three visits; both the right and left eyes of the patients were examined during these visits. Each observer calculated inter-observer and intra-observer variation as intraclass correlation coefficient (ICC). To determine the reliability, Cronbach's alpha was calculated. A P value of 0.9, and the overall ICC was also >0.9. The results showed excellent inter- and intra-observer reliability for the observers. Conclusion: The IOP values measured using the GAT were found to have significant intra- and inter-observer reliability. This demonstrates that ophthalmologists can perform valid and reliable IOP assessments. This also indicates the importance of careful and proper training of observers in measuring IOP using the GAT and the standardisation of criteria for early detection of glaucoma

FORMULATION AND CHARACTERIZATION OF LOSARTAN LOADED SELF EMULSIFYING DRUG DELIVERY SYSTEM

Objective: The object of the present work was the advancement and portrayal of losartan stacked self-emulsifying drug conveyance framework for the successful administration of hypolipidemia (RSEDDS) for further developing bioavailability, to upgrade dissolvability, delays home time, give an adequate measure of medication to an objective site and supported the arrival of medication. Methods: Self-Emulsifying Drug Delivery System was ready by the basic emulsification method. Six clumps, for example, F1 to F6 were ready by shifting the convergence of oils, surfactant, co-surfactant, and co-dissolvable and assessed for the different boundaries, for example, Optical microscopy, Assessment of self emulsification, Emulsification time, Droplet size investigation, Zeta Potential Measurement, Transmission Electron Microscopy, Viscosity Determination, Drug content, Percentage conveyance, in vitro disintegration study and solidness study. The SEDDS was upgraded and group F5 was additionally utilized. Results: The medication content of chosen clump F5 was viewed as 97.65±1.37 %; it proposes that the technique for exemplification was powerful. As per in vitro study, around 55.13 % of the medication was delivered after 120 min which showed supported arrival of medication and there were no critical changes seen in the actual appearance, drug content, and in vitro drug release during the stability study. Conclusion: This research presumed that the SEDDS are an expected competitor as a supported delivery drug conveyance, effectively expanding bioavailability and designated conveyance of medication

Divergent Roles of Escherichia Coli Encoded Lon Protease in Imparting Resistance to Uncouplers of Oxidative Phosphorylation:Roles of marA, rob, soxS and acrB

Uncouplers of oxidative phosphorylation dissipate the proton gradient, causing lower ATP production. Bacteria encounter several non-classical uncouplers in the environment, leading to stress-induced adaptations. Here, we addressed the molecular mechanisms responsible for the effects of uncouplers in Escherichia coli. The expression and functions of genes involved in phenotypic antibiotic resistance were studied using three compounds: two strong uncouplers, i.e., Carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 2,4-Dinitrophenol (DNP), and one moderate uncoupler, i.e., Sodium salicylate (NaSal). Quantitative expression studies demonstrated induction of transcripts encoding marA, soxS and acrB with NaSal and DNP, but not CCCP. Since MarA and SoxS are degraded by the Lon protease, we investigated the roles of Lon using a lon-deficient strain (Δlon). Compared to the wild-type strain, Δlon shows compromised growth upon exposure to NaSal or 2, 4-DNP. This sensitivity is dependent on marA but not rob and soxS. On the other hand, the Δlon strain shows enhanced growth in the presence of CCCP, which is dependent on acrB. Interestingly, NaSal and 2,4-DNP, but not CCCP, induce resistance to antibiotics, such as ciprofloxacin and tetracycline. This study addresses the effects of uncouplers and the roles of genes involved during bacterial growth and phenotypic antibiotic resistance. Strong uncouplers are often used to treat wastewater, and these results shed light on the possible mechanisms by which bacteria respond to uncouplers. Also, the rampant usage of some uncouplers to treat wastewater may lead to the development of antibiotic resistance.</p

Role of Human Epididymis Protein 4 in Tumour Angiogenesis

HE4 is a secretory protein. It is expressed in reproductive tract and respiratory epithelium in normal individuals. Serum level of HE4 is raised in various solid cancers that give us an advantage to use it as a diagnostic and prognostic biomarker. It is an established biomarker of epithelial ovarian cancer [EOC]. It has also shown the significance in various other malignancies like cancer of endometrium, cervix, lung and breast. Studies show HE4 as an independent prognostic biomarker in non-small cell lung carcinoma. Its raised values in cancer signify its role in oncogenesis. HE4 promotes angiogenesis via STAT3 signalling pathway. In this paper we have tried to illustrate about human epididymis protein 4 and its role in tumour angiogenesis

Evaluation of HE4 as a prognostic biomarker in uterine cervical cancer,

Introduction: Uterine cervical cancer (UCC) is the fourth most common health problem worldwide among women. Currently available biomarkers CA125, CA199, and CEA for diagnosis or prognostic evaluation of UCC have not got widespread acceptance. Method: Whole blood samples of 64 patients with UCC were collected along with 63 healthy females and tested for serum levels of HE4 (sHE4). A cut-off value for positive result 64.0 pmol/L was set. Statistical analysis of different clinical variables was done. Result: Serum level of HE4 has a significant role in the diagnosis of uterine cervical cancer. Its level increases with age, higher parity (P < 0.05), stage (P < 0.16), tumor size, and parametrial invasion. Negative result was seen with vaginal invasion, lymph node involvement &amp; cases which had recurrence. Various histological types showed variable results. So the serum level of HE4 (sHE) level may play a role in the diagnosis &amp; therapeutic monitoring of UCC. But the prognostic evaluation needs further studies. Conclusion: sHE4 is useful in the diagnosis of cervical cancer, but its prognostic significance is under the question marks. It may be associated with higher values in higher stages. Higher parity of the patient is associated with higher level of HE4 in UCC