Blood Cell Profiles of the Tadpoles of the Dubois's Tree Frog, Polypedates teraiensis Dubois, 1986 (Anura: Rhacophoridae)

The present paper describes a sequential study of the leukocyte profiles and the changes in morphometry and morphology of erythrocytes in the tadpoles of Polypedates teraiensis during their development and metamorphosis, that is, transfer from an aquatic mode to a terrestrial mode of life. Blood smears of 21 different stages (Gosner stage 26 to 46) of tadpoles were investigated. Population of erythrocytes was heterogeneous in population represented by various forms (oval, elliptical or rounded cells, comma shaped, teardrop shaped, schistocytes, senile erythrocytes, crenulated RBCs). Correlation between various morphometric values of erythrocytes was determined with different developing stages of tadpoles. Amongst the leucocytes, the lymphocytes were the most abundant cells followed by neutrophils. Neutrophils and monocytes showed varied morphologic forms. The percentage of lymphocytes and neutrophils showed a negative whereas percentage of eosinophil, basophil, and monocytes showed a positive correlation with the developmental stages of tadpoles. Blood platelets were also observed, which were rounded in shape and found in aggregates

Hematology of Wild Caught Dubois’s Tree Frog Polypedates teraiensis

Blood was analyzed from eighty (forty males and forty females) adult individuals of Polypedates teraiensis to establish reference ranges for its hematological and serum biochemical parameters. The peripheral blood cells were differentiated as erythrocytes, lymphocytes, eosinophils, neutrophils, monocytes, basophils, and thrombocytes, with similar morphology to other anurans. Morphology of blood cells did not vary according to sex. The hematological investigations included morphology and morphometry of erythrocytes, morphometry of leucocytes, packed cell volume (PCV), hemoglobin content (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), erythrocyte or red blood cell (RBC) count, leukocyte or white blood cell (WBC) count, differential leukocyte count, and neutrophil to lymphocyte (N/L) ratio. Besides, protein, cholesterol, glucose, urea, uric acid, and creatinine content of blood serum were assayed. Hematological parameters that differed significantly between sexes were RBC count, length and breadth of RBC, neutrophil %, N/L ratio, area occupied by basophils, and diameter of large lymphocyte and eosinophils. The level of glucose, urea, and creatinine in blood serum also significantly differed between sexes

Perspectives of traditional health care system of Sikkim, North-East India – Anethno-pharmacological survey and analysis

The Indo-Himalayan mountain state of Sikkim has abundant reserves of bio-diversity of ancient ancestry. Many folklore healers renowned in Sikkim for their valuable traditional knowledge, especially for the use of combination drug therapy to treat bone fracture. In the cross-cultural ethnopharmacological survey, a predesigned questionnaire was used for interviews at the residence of respective folklore healers. Their patient handling and preparations of formulations have documented in written and audio-visual format. The standard statistical indices selected relevant for the present study. The record of traditional knowledge on 193 different formulations used for 49 various human ailments has enumerated during this survey. A total of 121 plants belonging to 65 families were found to be used as a component for 193 formulations. Graphical representation of the frequency of citation, especially survey and reference data shown significant correlation indicating common and specific use of plants in the treatment of different diseases. The scientific research on medicinal plants used by traditional practitioners and the application of ethnobotanical products in the folklore healthcare system significantly help in the sustainable development of traditional healthcare practices of the region. For the research fraternity, the quantitative analysis of survey data is gaining wider acceptability due to relative importance

Perspectives of traditional health care system of Sikkim, North-East India – An ethno-pharmacological survey and analysis

965-981The Indo-Himalayan Mountain state of Sikkim has abundant reserves of bio-diversity of ancient ancestry. Many folklore healers are renowned in Sikkim for their valuable traditional knowledge, especially for the use of combination drug therapy to treat bone fracture. In the cross-cultural ethnopharmacological survey, a predesigned questionnaire was used for interviews at the residence of respective folklore healers. Their patient handling and preparations of formulations have been documented in written and audio-visual format. The standard statistical indices selected relevant for the present study. The record of traditional knowledge on 193 different formulations used for 49 various human ailments have been enumerated during this survey. A total of 121 plants belonging to 65 families were found to be used as a component for 193 formulations. Graphical representation of the frequency of citation, especially survey and reference data shown significant correlation indicating common and specific use of plants in the treatment of different diseases. The scientific research on medicinal plants used by traditional practitioners and the application of ethnobotanical products in the folklore healthcare system significantly help in the sustainable development of traditional healthcare practices of the region. For the research fraternity, the quantitative analysis of survey data is gaining wider acceptability due to relative importance

Detection of Target DNA using Photo-Reactive Protoporphyrin Moeity on a Nanocomposite Substrate

Detection of pathogens from infected biological samples through conventional process involves cell lysis and purification. The main objective of this work is to minimize the time and sample loss, as well as to increase the efficiency of detection of biomolecules. Electrical lysis of medical sample is performed in a closed microfluidic channel in a single integrated platform where the downstream analysis of the sample is possible. The device functions involve, in a sequence, flow of lysate from lysis chamber passed through a thermal denaturation counter where dsDNA is denatured to ssDNA, which is controlled by heater unit. A functionalized binding chamber of ssDNA is prepared by using ZnO nanorods as the matrix and functionalized with bifunctional carboxylic acid, 16-(2-pyridyldithiol) hexadecanoic acid (PDHA) which is further attached to a linker molecule 1-ethyl-3-(3-dimethylaminopropyl) (EDC). Linker moeity is then covalently bound to photoreactive protoporphyrin (PPP) molecule. The photolabile molecule protoporphyrin interacts with -NH2 labeled single stranded DNA (ssDNA) which thus acts as a probe to detect complimentary ssDNA from target organisms. Thereafter the bound DNA with protoporphyrin is exposed to an LED of particular wavelength for a definite period of time and DNA was eluted and analyzed. UV/Vis spectroscopic analysis at 260/280 nm wavelength confirms the purity and peak at 260 nm is reconfirmed for the elution of target DNA. Quantitative and qualitative data obtained from the current experiments show highly selective detection of biomolecule such as DNA which have large number of future applications in Point-of-Care devices

MECHANISM OF CELL LYSIS IN MICROFLUIDIC CHANNEL WITH INTEGRATED NANOCOMPOSITE ELECTRODES

This paper reports on the characterization of an integrated micro-fluidic platform for controlled electrical lysis of biological cells and subsequent extraction of intracellular biomolecules. The proposed methodology is capable of high throughput electrical cell lysis facilitated by nano-composite coated electrodes. The nano-composites are synthesized using Carbon Nanotube and ZnO nanorod dispersion in polymer. Bacterial cells are used to demonstrate the lysis performance of these nanocomposite electrodes. Investigation of electrical lysis in the microchannel is carried out under different parameters, one with continuous DC application and the other under DC biased AC electric field. Lysis in DC field is dependent on optimal field strength and governed by the cell type. By introducing the AC electrical field, the electrokinetics is controlled to prevent cell clogging in the micro-channel and ensure uniform cell dispersion and lysis. Lysis mechanism is analyzed with time-resolved fluorescence imaging which reveal the time scale of electrical lysis and explain the dynamic behavior of GFP-expressing E. coli cells under the electric field induced by nanocomposite electrodes. The DNA and protein samples extracted after lysis are compared with those obtained from a conventional chemical lysis method by using a UV-Visible spectroscopy and fluorimetry. The paper also focuses on the mechanistic understanding of the nano-composite coating material and the film thickness on the leakage charge densities which lead to differential lysis efficiency

Charge Injection through Nanocomposite Electrode in Microfluidic Channel for Electrical Lysis of Biological Cells

Several concepts have been developed in the recent years for nanomaterial based integrated MEMS platform in order to accelerate the process of biological sample preparation followed by selective screening and identification of target molecules. In this context, there exist several challenges which need to be addressed in the process of electrical lysis of biological cells. These are due to (i) low resource settings while achieving maximal lysis (ii) high throughput of target molecules to be detected (iii) automated extraction and purification of relevant molecules such as DNA and protein from extremely small volume of sample (iv) requirement of fast, accurate and yet scalable methods (v) multifunctionality toward process monitoring and (vi) downward compatibility with already existing diagnostic protocols. This paper reports on the optimization of electrical lysis process based on various different nanocomposite coated electrodes placed in a microfluidic channel. The nanocomposites are synthesized using different nanomaterials like Zinc nanorod dispersion in polymer. The efficiency of electrical lysis with various different electrode coatings has been experimentally verified in terms of DNA concentration, amplification and protein yield. The influence of the coating thickness on the injection current densities has been analyzed. We further correlate experimentally the current density vs. voltage relationship with the extent of bacterial cell lysis. A coupled multiphysics based simulation model is used to predict the cell trajectories and lysis efficiencies under various electrode boundary conditions as estimated from experimental results. Detailed in-situ fluorescence imaging and spectroscopy studies are performed to validate various hypotheses

Perspectives of traditional health care system of Sikkim, North-East India – An ethno-pharmacological survey and analysis

965-981The Indo-Himalayan Mountain state of Sikkim has abundant reserves of bio-diversity of ancient ancestry. Many folklore healers are renowned in Sikkim for their valuable traditional knowledge, especially for the use of combination drug therapy to treat bone fracture. In the cross-cultural ethnopharmacological survey, a predesigned questionnaire was used for interviews at the residence of respective folklore healers. Their patient handling and preparations of formulations have been documented in written and audio-visual format. The standard statistical indices selected relevant for the present study. The record of traditional knowledge on 193 different formulations used for 49 various human ailments have been enumerated during this survey. A total of 121 plants belonging to 65 families were found to be used as a component for 193 formulations. Graphical representation of the frequency of citation, especially survey and reference data shown significant correlation indicating common and specific use of plants in the treatment of different diseases. The scientific research on medicinal plants used by traditional practitioners and the application of ethnobotanical products in the folklore healthcare system significantly help in the sustainable development of traditional healthcare practices of the region. For the research fraternity, the quantitative analysis of survey data is gaining wider acceptability due to relative importance

Infection-Related Ventilator-Associated Complication and Possible Ventilator-Associated Pneumonia among Mechanically Ventilated Patients of Adult Medical and Surgical Intensive Care Units

Objective An observational study was conducted to evaluate (1) the incidence rates of infection-related ventilator-associated complication (IVAC) and possible ventilator-associated pneumonia (PVAP) among mechanically ventilated patients of adult medical and surgical intensive care units (ICUs) and (2) the pathogen distribution in patients with PVAP. Materials and Methods The IVAC and PVAP rates of medical and surgical ICUs, between July 1, 2017, and June 30, 2021, per 1,000 mechanical ventilator (MV) days were calculated. The significance of difference in IVAC and PVAP rates between medical and surgical ICUs was calculated. The level of significance was set at less than 0.05. Results MV utilization ratios of adult medical and surgical ICUs were 0.32 and 0.26, respectively (p  0.05) and PVAP rates of 2.46 and 1.59 per 1,000 MV days in medical and surgical ICUs, respectively (p > 0.05). Acinetobacter baumannii complex either singly or in combination was isolated in 11/20 PVAP cases. Conclusion IVAC and PVAP were more in medical compared with surgical ICUs. The most common pathogen in patients with PVAP was A. baumannii complex. More studies are warranted to monitor the significance of ventilator-associated event on patient outcomes