Histological, Histochemical and Biochemical characterisation of male morphotypes of Macrobrachium rosenbergii (de Man)

This thesis entitled “Histological ,Histochemical and biochemical characterisation of male morphotypes of macrobrachium rosenbergii(De Man). The giant fresh water prawn Macrobrachium rosenbergii (de Man) is emerging as a prime candidate species in fresh water aquaculture as a global basis and therefore, receiving much attention in recent years.the present work was aimed at to study the histological variations, if any, in the reproductive system viz. testes, vas deferens including androgenic gland, hepatopancreas and the neurosecretory system viz. eye stalk, brain and thoracic ganglion among the male morphotypes and their transitional stages of M.rosenbergii from growouts. This study was also aimed at to bring out the histochemical variations, if any, in the reproductive system comprising of testes, vas deferens including androgenic gland and the hepatopancreas among the male morphotypes and their transitional stages collected from growouts. Biochemical characterisation of various male morphotyes and their transitional stages have also been attempted in order to find out biochemical evidence, if any, in the morphotypic transformation.Histological study of the testes of three male morphotypes viz., SM, SOC and SBC and their 'transitional stages viz., WOC, tSOC, WBC and OBC have been carried out with a view to unravel the structural and functional differences of the testes, if any, of these morphotypes. Studies on the lipid components viz., cholesterol, phospholipid and triglyceride In the muscle tissue and hepatopancreas have been carried out.

Studies on the histological and biochemical changes during spermatogenesis in MUGIL CEPHALUS linnaeus and related species

Several studies on the biology and fisheries of mullets, particularly of M cephalus are now available. Different aspects of breeding, larval rearing, seed production, field culture and ecophysiology have also been investigated. However, information on the spermatogenesis in M cephalus as well as L parsia is scanty. Since an understanding of the reproductive strategies is an essential pre-requisite for evolving successful breeding programmes through artificial fertilization and gametic preservation, investigations on spermatogenesis in these species were taken up and the results are presented in this thesis. The thesis is presented in 9 chapters

Source Characterization of Sedimentary Organic Matter in a Tropical Estuary, Southwest Coast of India: A Biomarker Approach

The source, fate and diagentic pathway of sedimentary organic matter in estuaries are difficult to delineate due to the complexity of organic matter sources, intensive physical mixing and biological processes. A combination of bulk organic matter techniques and molecular biomarkers are found to be successful in explaining organic matter dynamics in estuaries. The basic requirement for these multi-proxy approaches are (i) sources have significantly differing characteristics, (ii) there are a sufficient number of tracers to delineate all sources and (iii) organic matter degradation and processing have little, similar or predictable effects on end member characteristics. Although there have been abundant researches that have attempted to tackle difficulties related to the source and fate of organic matter in estuarine systems, our understanding remains limited or rather inconsistent regarding the Indian estuaries. Cochin estuary is the largest among many extensive estuarine systems along the southwest coast of India. It supports as much biological productivity and diversity as tropical rain forests. In this study, we have used a combination of bulk geochemical parameters and different group of molecular biomarkers to define organic matter sources and thereby identifying various biogeochemical processes acting along the salinity gradient of the Cochin estuary

Biomarker pigment signatures in Cochin back water system e A tropical estuary south west coast of India

Sedimentary biomarker pigments around Cochin estuary situated in the southwest coast of India were determined by HPLC. Fucoxanthin, an indicator of diatom was observed to be the most abundant carotenoid pigment in the estuary. Dinoflagellate derived carotenoid pigment peridinin was confined in the southern part of estuary and zeaxanthin pigment indicative of cyanobacteria were more found in sites influenced by anthropogenic activities. One compound having close similarity to fucoxanthin was also detected. Alloxanthin (cryptophyceae), chl b (green algae), canthaxanthin, neoxanthin, lutein and peridinin isomer were also detected by spectra and corresponding algal class were identified. The highest concentration of chl a (11.01 mg g 1) found near to the anthropogenic affected area while the lowest chl a (0.65 mg g 1) was recorded in industrial area. Degradation products of chl a, such as pheophorbide and pheophytin were observed and principal mode of mechanism of degradation were derived. Higher pheopigments content than chl a, reflects a density trapping of dead cells and early degradation of phytopigments from grazing activities

Protein-Expressions-Analysen zur Etablierung prädiktiver Biomarker bei der Behandlung fortgeschrittener Tumoren der Mamma und der Prostata

Aktuelle Entwicklungen auf dem Gebiet der zielgerichteten Therapie zur Behandlung maligner Erkrankungen erfordern neuartige Verfahren zur Diagnostik und Selektion geeigneter Patienten. So ist das Ziel der vorliegenden Arbeit die Identifizierung neuer Zielmoleküle, die die Vorhersage eines Therapieerfolges mit targeted drugs ermöglichen. Besondere Aufmerksamkeit gilt dem humanisierten monoklonalen Antikörper Trastuzumab (Herceptin), der zur Therapie Her-2 überexprimierender, metastasierter Mammakarzinome eingesetzt wird. Jüngste Erkenntnisse lassen eine Anwendung dieses Medikamentes in der Behandlung des Hormon-unabhängigen Prostatakarzinoms möglich erscheinen. Therapie-beeinflussende Faktoren werden in der dem Rezeptor nachgeschalteten Signaltransduktion oder Veränderungen des Rezeptors selbst vermutet. Mittels Immunhistochemie wurden die Expressions- und Aktivierungsniveaus verschiedener Proteine der Her-2-assoziierten Signaltransduktion ermittelt; insgesamt wurden 37 molekulare Marker untersucht. In Formalin fixierte und in Paraffin eingebettete korrespondierende Normal- und Tumorgewebe von 118 Mammakarzinom-Patientinnen sowie 78 Patienten mit Prostatakarzinom wurden in TMAs zusammengefasst. Die in Zusammenarbeit mit erfahrenen Pathologen ermittelten Ergebnisse dienten u.a. als Grundlage für zweidimensionales, unsupervised hierarchisches clustering. Ergebnis dieser Analysen war für beide untersuchten Tumorentitäten die Möglichkeit einer Subklassifizierung der untersuchten Populationen nach molekularen Eigenschaften. Hierbei zeigten sich jeweils neue Möglichkeiten zur Anwendung zielgerichteter Therapien, deren Effektivität Inhalt weiterführender Studien sein könnte. Zusätzlich wurden an insgesamt 43 Frischgeweben die möglichen Folgen des sog. shedding untersucht. Western Blot-basierte Untersuchungen zeigten hierbei die Möglichkeit der Selektion von Patienten aufgrund falsch-positiver Befunde in der derzeit als Standard geltenden Diagnostik. Zusätzlich konnte durch Vergleich mit einer Herceptin-sensitiven Zelllinie ein möglicher Zusammenhang eines Therapieerfolges mit dem Phosphorylierungs-/ Aktivierungszustand des Rezeptors ermittelt werden. Fehlende klinische Daten zum Verlauf der Erkrankung und Therapie der untersuchten Patienten lassen keine Aussagen über die tatsächliche Relevanz der ermittelten Befunde zu. Dennoch verdeutlichen die erhaltenen Resultate eindrucksvoll die Komplexität der molekularen Vorgänge, die zu einem Krebsgeschehen führen und damit Auswirkungen auf die Wirksamkeit von targeted drugs haben können. Entwicklungen auf dem Gebiet der zielgerichteten Therapie erfordern Verbesserungen auf dem Gebiet der Diagnostik, die die sichere Selektion geeigneter Patienten erlauben. Die Zukunft der personalisierten, zielgerichteten Behandlung von Tumorerkrankungen wird verstärkt von molekularen Markerprofilen hnlich den hier vorgestellten Daten beeinflusst werden.

Carga viral de seis tipos de Virus del Papiloma Humano de alto riesgo y su asociacion con lesiones cervicales

Introducción: La infección por un tipo de Virus del Papiloma Humano de alto riesgo (VPH-AR), es el factor principal en el desarrollo de Cáncer de Cérvix (CC). La carga viral puede modular esta asociación, por lo que resulta importante su cuantificación y el establecimiento de su relación con lesiones precursoras de CC. Metodología: 60 mujeres con lesiones escamosas intraepiteliales (LEI) y 120 mujeres sin LEI, confirmadas por colposcopia, fueron incluidas en el estudio. Se determinó la carga viral de 6 tipos de VPH-AR, mediante PCR en tiempo real. Se estimaron OR crudos y ajustados para evaluar la asociación entre la carga viral de cada tipo y las lesiones cervicales. Resultados: 93.22% de mujeres con LEI y 91.23% de mujeres negativas, fueron positivas para al menos un tipo de VPH. VPH-18 y VPH-16 fueron los tipos más prevalentes, junto con VPH-31 en mujeres sin LEI. No se encontraron diferencias estadísticamente significativas de las cargas virales entre éstos dos grupos, aunque se observó un mayor carga viral en lesiones para algunos tipos virales. Una mayor frecuencia de lesiones se asoció a infecciones con carga baja de VPH-16 (ORa: 3.53; IC95%: 1.16 – 10.74), en comparación a mujeres con carga alta de VPH-16, (ORa: 2.63; IC95%: 1.09 – 6.36). En infecciones por VPH-31, la presencia de carga viral alta, se asoció con una menor frecuencia de lesiones (ORa: 0.34; IC95%: 0.15 – 0.78). Conclusiones: La prevalencia tipo-específica de VPH se corresponde con las reportadas a nivel mundial. La asociación entre la carga viral del VPH y la frecuencia de LEI es tipo específica y podría depender de la duración de la infección, altas cargas relacionadas con infecciones transitorias, y bajas cargas con persistentes. Este trabajo contribuye al entendimiento del efecto de la carga viral en la historia natural del CC; sin embargo, estudios prospectivos son necesarios para confirmar estos resultados.

Histological and functional aspects of Alpha-synuclein in neurodegeneration

Alpha-synuclein has been implicated in the cellular mechanisms that control auditory sensitivity. In other systems it can also confer protection against cellular injury. Auditory brainstem response thresholds and immunohistochemistry were used to assess the ability of alpha-synuclein to protect against oxidative damage to the cochlea.

Verification of computed tomographic estimates of cochlear implant array position: A micro-CT and histological analysis

We examine the efficacy two volume spatial registration of pre and postoperative clinical computed tomography (CT) imaging to verify post-operative electrode array placement in cochlear implant (CI) patients. To measure the degree of accuracy with which the composite image predicts in-vivo placement of the array, we replicate the CI surgical process in cadaver heads. Pre-operative, post-operative, micro CT imaging and histology are utilized for verification.

Effect of sex and genotype on cardiovascular biomarker response to fish oils: the FINGEN Study

Background: The lipid-modulatory effects of high intakes of the fish-oil fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are well established and likely to contribute to cardioprotective benefits. Objectives: We aimed to determine the effect of moderate EPA and DHA intakes (< 2 g EPA + DHA/d) on the plasma fatty acid profile, lipid and apolipoprotein concentrations, lipoprotein subclass distribution, and markers of oxidative status. We also aimed to examine the effect of age, sex, and apolipoprotein E (APOE) genotype on the observed responses. Design: Three hundred twelve adults aged 20-70 y, who were prospectively recruited according to age, sex, and APOE genotype, completed a double-blind placebo-controlled crossover study. Participants consumed control oil, 0.7 g EPA + DHA/d (0.7FO), and 1.8 g EPA + DHA/d (1.8FO) capsules in random order, each for an 8-wk intervention period, separated by 12-wk washout periods. Results: In the group as a whole, 8% and 11% lower plasma triacylglycerol concentrations were evident after 0.7FO and 1.8FO, respectively (P < 0.001): significant sex x treatment (P = 0.038) and sex x genotype x treatment (P = 0.032) interactions were observed, and the greatest triacylglycerol-lowering responses (reductions of 15% and 23% after 0.7FO and 1.8FO, respectively) were evident in APOE4 men. Furthermore, lower VLDL-cholesterol (P = 0.026) and higher LDL-cholesterol (P = 0.010), HDL-cholesterol (P < 0.001), and HDL2 (P < 0.001) concentrations were evident after fish-oil intervention. Conclusions: Supplements providing EPA + DHA at doses as low as 0.7 g/d have a significant effect on the plasma lipid profile. The results of the current trial, which used a prospective recruitment approach to examine the responses in population subgroups, are indicative of a greater triacylglycerol-lowering action of long-chain n-3 polyunsaturated fatty acids in males than in females.

Haem peroxidase activity in Daphnia magna: A biomarker for sub-lethal toxicity assessments of kerosene-contaminated groundwater

A novel biomarker was developed in Daphnia magna to detect organic pollution in groundwater. The haem peroxidase assay, which is an indirect means of measuring oxidase activity, was particularly sensitive to kerosene contamination. Exposure to sub-lethal concentrations of kerosene-contaminated groundwater resulted in a haem peroxidase activity increase by dose with a two-fold activity peak at 25%. Reproduction in D. magna remained unimpaired when exposed to concentrations below 25% for 21 days, and a decline in fecundity was only observed at concentrations above the peak in enzyme activity. The measurement of haem peroxidase activity in D. magna detected sublethal effects of kerosene in just 24 h, whilst offering information on the health status of the organisms. The biomarker may be useful in determining concentrations above which detrimental effects would occur from long-term exposure for fuel hydrocarbons. Moreover, this novel assay detects exposure to chemicals in samples that would normally be classified as non-toxic by acute toxicity tests.

Incorporation of in situ and biomarker assays in higher-tier assessment of the aquatic toxicity of insecticides

This study was designed to test the feasibility of integrating in situ, single species exposures and biomarker analysis into microcosm studies. Experimental ponds were dosed with pirimiphos methyl (PM) and lindane. C. riparius fourth instar larvae were deployed for 48 h on nine separate occasions during the study period before and after treatment. Surviving larvae were analysed for acetylcholinesterase activity (AChE). Survival and biomarker data were compared to chironomid assemblage analysis by monitoring insects emerging from the microcosms. Survival of chironomids within the in situ systems commenced on day + 16 after treatment with 31.6% and 53.3% survival in the lindane and PM treated ponds, respectively. In contrast, the first emergence from the microcosms occurred on days + 27, in respect to lindane, and + 59 for the PM treated ponds. Thus the in situ bioassay was able to demonstrate gradual reduction in toxicity within the sediment before this was evident from macroinvertebrate monitoring. Significant ACNE inhibition was only detected on exposure to PM. Levels decreased from 75% on day + 16 to 26% by day +29. The biomarker analysis confirmed that, by the end of the study, the insecticide was no longer exerting an effect. We discuss how the use of in situ bioassays could also aid comparison of microcosm studies by adding a standardized dimension. (C) 2003 Elsevier Ltd. All rights reserved.

Intraclonal variability in Daphnia acetylcholinesterase activity: The implications for its applicability as a biomarker

The relationship between individual growth and acetylcholinesterase (AChE).activity was evaluated for Daphnia magna. Analysis on the influence of two different culture media on baseline AChE activity was performed with Daphnia similis. The results indicated an inverse relationship between D. magna body length and AChE activity. An increase in total protein, which was not proportional to an increase in the rate of the substrate hydrolysis (Delta absorbance/min), seems to be the reason for this inverse size versus AChE activity relationship. Therefore, toxicants such as phenobarbital, which affect protein and size but not AChE activity directly, have an overall affect on AChE activity. In contrast, the AChE inhibitor parathion altered AChE activity but not protein. Culture medium also had a significant affect on AChE activity in D. similis. Changes in total protein seem to be the main reason for the variations in baseline AChE activity in Daphnia observed in the different evaluations performed in this work. Therefore, AChE activity in Daphnia must be interpreted carefully, and variations related to changes in total protein must be taken into account when applying this enzyme as a biomarker in biological monitoring.

Diet, fecal water, and colon cancer - development of a biomarker

Colorectal cancer (CRC) is a leading cause of cancer incidence worldwide. Lifestyle factors, especially dietary intake, affect the risk of CRC development. Suitable risk biomarkers are required in order to assess the effect that specific dietary components have on CRC risk. The relationship between dietary intake and indicators of fecal water activity has been assessed using cell and animal models as well as human studies. This review summarizes the literature on fecal water and dietary components with a view to establishing further the potential role of fecal water as a source of CRC risk biomarkers. The literature indicates that fecal water activity markers are affected by specific dietary components linked with CRC risk: red meat, saturated fats, bile acids, and fatty acids are associated with an increase in fecal water toxicity, while the converse appears to be true for calcium, probiotics, and prebiotics. However, it must be acknowledged that the study of fecal water is still in its infancy and a number of issues need to be addressed before its usefulness can be truly gauged.

Fecal water as a non-invasive biomarker in nutritional intervention: comparison of preparation methods and refinement of different endpoints

The assessment of cellular effects by the aqueous phase of human feces (fecal water, FW) is a useful biomarker approach to study cancer risks and protective activities of food. In order to refine and develop the biomarker, different protocols of preparing FW were compared. Fecal waters were prepared by 3 methods: (A) direct centrifugation; (B) extraction of feces in PBS before centrifugation; and (C) centrifugation of lyophilized and reconstituted feces. Genotoxicity was determined in colon cells using the Comet assay. Selected samples were investigated for additional parameters related to carcinogenesis. Two of 7 FWs obtained by methods A and B were similarly genotoxic. Method B, however, yielded higher volumes of FW, allowing sterile filtration for long-term culture experiments. Four of 7 samples were non-genotoxic when prepared according to all 3 methods. FW from lyophilized feces and from fresh samples were equally genotoxic. FWs modulated cytotoxicity, paracellular permeability, and invasion, independent of their genotoxicity. All 3 methods of FW preparation can be used to assess genotoxicity. The higher volumes of FWobtained by preparation method B greatly enhance the perspectives of measuring different types of biological parameters and using these to disclose activities related to cancer development.

Effect of sex and genotype on cardiovascular biomarker response to fish oils: the FINGEN Study

Background: The lipid-modulatory effects of high intakes of the fish-oil fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are well established and likely to contribute to cardioprotective benefits. Objectives: We aimed to determine the effect of moderate EPA and DHA intakes (< 2 g EPA + DHA/d) on the plasma fatty acid profile, lipid and apolipoprotein concentrations, lipoprotein subclass distribution, and markers of oxidative status. We also aimed to examine the effect of age, sex, and apolipoprotein E (APOE) genotype on the observed responses. Design: Three hundred twelve adults aged 20-70 y, who were prospectively recruited according to age, sex, and APOE genotype, completed a double-blind placebo-controlled crossover study. Participants consumed control oil, 0.7 g EPA + DHA/d (0.7FO), and 1.8 g EPA + DHA/d (1.8FO) capsules in random order, each for an 8-wk intervention period, separated by 12-wk washout periods. Results: In the group as a whole, 8% and 11% lower plasma triacylglycerol concentrations were evident after 0.7FO and 1.8FO, respectively (P < 0.001): significant sex x treatment (P = 0.038) and sex x genotype x treatment (P = 0.032) interactions were observed, and the greatest triacylglycerol-lowering responses (reductions of 15% and 23% after 0.7FO and 1.8FO, respectively) were evident in APOE4 men. Furthermore, lower VLDL-cholesterol (P = 0.026) and higher LDL-cholesterol (P = 0.010), HDL-cholesterol (P < 0.001), and HDL2 (P < 0.001) concentrations were evident after fish-oil intervention. Conclusions: Supplements providing EPA + DHA at doses as low as 0.7 g/d have a significant effect on the plasma lipid profile. The results of the current trial, which used a prospective recruitment approach to examine the responses in population subgroups, are indicative of a greater triacylglycerol-lowering action of long-chain n-3 polyunsaturated fatty acids in males than in females.