Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions. The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of human serum suggesting liver damage. This was also confirmed by histopathological studies.
SmartScreen Technology for Building a Better AssayKristin Rider
A lipid derived nanoparticle the recreates the cellular membrane in solution based assays. See increased enzymatic activity, identify more relevant biological substrates, find novel hits from the compound library.
Poster demonstrating the results from the development/verification project for the quantitation of pyridoxal 5-phosphate and 4-pyridoxic acid in human plasma.
Synthesis, spectral characterization and bioactivity studies of some S-substi...Jing Zang
A new series of 5-(4-Chlorophenyl)-1,3,4-Oxadiazol-2-thiol derivatives was prepared from 4-chlorobenzoic acid (1) by converting it successively into corresponding ester (2), carbohydrazide (3) and 5-(4-Chlorophenyl)-1,3,4-Oxadiazol-2-thiol (4). Finally the target compounds, 6a-l, were synthesized by stirring 4 with different electrophiles, 5a-l, in DMF using NaH as weak base and activator. The proposed structures of newly synthesized compounds were confirmed by spectroscopic techniques such as 1H-NMR, 13C-NMR, HR-MS and EI-MS. All synthesized compounds were evaluated for their anti-bacterial, antifungal, cytotoxicity and enzyme inhibition activities. The compounds, 6e and 6g exhibited significant inhibition activity against acetyl cholinesterase enzyme (AChE) and 6j moderate activity against butyryl cholinesterase enzyme (BChE). The molecule, 4 exhibited good MIC (minimum inhibitory concentration) value against all the bacterial and fungal strains taken into account.
SmartScreen Technology for Building a Better AssayKristin Rider
A lipid derived nanoparticle the recreates the cellular membrane in solution based assays. See increased enzymatic activity, identify more relevant biological substrates, find novel hits from the compound library.
Poster demonstrating the results from the development/verification project for the quantitation of pyridoxal 5-phosphate and 4-pyridoxic acid in human plasma.
Synthesis, spectral characterization and bioactivity studies of some S-substi...Jing Zang
A new series of 5-(4-Chlorophenyl)-1,3,4-Oxadiazol-2-thiol derivatives was prepared from 4-chlorobenzoic acid (1) by converting it successively into corresponding ester (2), carbohydrazide (3) and 5-(4-Chlorophenyl)-1,3,4-Oxadiazol-2-thiol (4). Finally the target compounds, 6a-l, were synthesized by stirring 4 with different electrophiles, 5a-l, in DMF using NaH as weak base and activator. The proposed structures of newly synthesized compounds were confirmed by spectroscopic techniques such as 1H-NMR, 13C-NMR, HR-MS and EI-MS. All synthesized compounds were evaluated for their anti-bacterial, antifungal, cytotoxicity and enzyme inhibition activities. The compounds, 6e and 6g exhibited significant inhibition activity against acetyl cholinesterase enzyme (AChE) and 6j moderate activity against butyryl cholinesterase enzyme (BChE). The molecule, 4 exhibited good MIC (minimum inhibitory concentration) value against all the bacterial and fungal strains taken into account.
Poster demonstrating the results from the development/verification project for the quantitation of all- trans retinol and alpha tocopherol in human serum.
Mycotoxins are are secondary metabolites produced by fungi and are dangerous for feed and food chains as they can create contamination in pre- and post-harvest processes. Many are highly toxic and as such levels in food products are regulated in Europe, the US, Japan and other countries. This presentation is an overview of the application of ultra-performance liquid chromatography combined with tandem quadrupole mass spectrometry to analyse various food products for mycotoxins in line with regulatory requirements.
Poster demonstrating the results from the development/verification project for the quantitation of all- trans retinol and alpha tocopherol in human serum.
Mycotoxins are are secondary metabolites produced by fungi and are dangerous for feed and food chains as they can create contamination in pre- and post-harvest processes. Many are highly toxic and as such levels in food products are regulated in Europe, the US, Japan and other countries. This presentation is an overview of the application of ultra-performance liquid chromatography combined with tandem quadrupole mass spectrometry to analyse various food products for mycotoxins in line with regulatory requirements.
Psychiatric Social Work Intervention for Social Rejection of persons with Sch...inventionjournals
It is widely acknowledged that schizophrenic patients have major deficits in social functioning in the community. The intervention to improve these deficits is inadequate in rural areas in India. The aim is to ascertain impact of structured psychiatric social work intervention for management of social rejection of persons with schizophrenia in rural areas. The sample consisted of thirty persons with schizophrenia and significant family members. A proportionate sample covering heterogeneous socio demographic background was drawn using simple random sampling technique. It was followed a Before-After Experimental Research Design Without Control Group. The social rejection and orientation towards mental illness were assessed before and after psychiatric social work intervention using Patient Rejection Scale and Orientation towards Mental Illness Scale. After psychiatric social work intervention overall there were changes in the domains of social rejection and orientation towards mental illness. The results have indicated that the psychiatric social work intervention has contributed to improve the social functioning of the patients in rural areas. Results have shown the changes in all the domains of social rejections and orientation towards mental illness significantly. Conducting similar studies in different settings would contribute to the psychiatric social work knowledge in future
la naturaleza es fuente de vida para todos los seres humanos porque sin ella todos nosotros estuviéramos muerto por eso debemos cuidarla y enseñar a ñas demás personas a que agan lo mismo y a que aprendan a no contaminar los ríos ni tampoco talar tantos árboles para tener un mundo mejor para que sea el ejemplo de otros lugares.
Adjudicacion herencia Luis Septien y Septien, YSITA DEL HOYOysitadelhoyo
En la escritura del legado del difunto Luis Septien no existe ninguna mención del Lic. FRANCISCO YSITA DEL HOYO ni tampoco del Lic. JORGE VILLATORO. Como testimonio ahi esta la escritura de su legado se hace mención al difunto Fernando Ysita Septien como albacea de la sucesión de Luis Septien, más no propietario de la fundación. Claramente se especifica que al fallecimiento de este pasara en primer lugar a la Arquidiocesis de Mexico sino a la Beneficencia Publica o la Cruz Roja, lo cual no sucedió.
Francisco Ysita del Hoyo estas lucreando con bienes que no son de tu propiedad. No abuses con supuestas ayuda comunitaria ya basta. Has logrado todo con tu íntimo amigo Notario Tomas Lozano Molina para utilizar poderes y escrituras apócrifas.
Citizen Satisfaction with Police: A Pillar of Law Enforcement Governanceinventionjournals
Increasing use of governance practices in public administration have had broad repercussions in law enforcement and policing. As a result, law enforcement services have witnessed significant changes that shifted police from a bureaucratic, incident driven force to a proactive service oriented organization. Among other governance indicators, citizen satisfaction and confidence in law enforcement services has increasingly been one of the central tenets of the governance structures in the contemporary world.In this new era of policing, enhancing community relations and improving the satisfaction of citizens have become the main objective of police departments. This study first explains the governance approach to law enforcement and policing and then focuses on citizen satisfaction with police. It explores the general determinants of citizen satisfaction, and assesses the impact of police citizen encounters on citizen satisfaction with police. Based on the literature and research studies, this article shows that socio-demographic characteristics, such as age, race, gender, and income independently determine the attitudes toward police. Police contact also has a significant effect on satisfaction level. Favorable contacts with police improve citizens’ satisfaction regardless of the type of contact. Residential location of an individual and physical police presence in the neighborhood also affects attitudes toward police.
Towards Indian Agricultural Information: A Need Based Information Flow Modelinventionjournals
Information is crucial for agriculture and rural development. ICT based services in agriculture is gaining importance day by day. Paper proposes to study the nature of agricultural information its uniqueness and problems of handling and organization. Agri informatics and use of different web portals for agriculture also discussed. Paper also highlighted the ICAR and IARIs contribution towards development of NARS (National Agricultural Research System) and models showing Information flow and strategic plan for organization of agricultural information. Some recommendations for proper organization and dissemination of agricultural information have been made to reach grass root level with desired agricultural information.
Fınancıal Rısk Management 'S Impact On Company Valueinventionjournals
The competition taking place under the current conditions reveals future uncertainty. To protect against possible adversities and create defenses at this point, no longer it has a compelling Taking requests from businesses. Financial risk management, reduction of the negative effects of the financial risks to the company, so the profitability of the company and the company is found to be useful to the extent it is important towards enhancing competitiveness. Financial risk management activities are also a cost themselves. The benefits will be provided for the management of financial risk, financial risk activities of the company are meaningless if they remain lower than the resources will be spent on risk management. However, the issue is quite complex, sectoral and disadvantages of return of financial risk management, and risk types can vary cyclical basis. international financial markets since the 1970s witnessed an important transition period. The globalization of international financial markets, free movement of capital, technological advances and increased competition a new process, launched the company with financial instability and has faced financial risk concept. Therefore institutions in ensuring the continuity of existence and reaching their goals were to identify financial risks, measure and manage today's extremely critical.
Culture of Renal Proximal Tubule Epithelial Cell Line SA7K Using Extracellula...mdmitc
MilliporeSigma's Toni Steiner recently presented a poster at the 2016 AAPS/ITC Transporter Workshop demonstrating how culture conditions can influence drug transporter expression and activity in renal proximal tubule epithelial cells.
Sachpazis:Terzaghi Bearing Capacity Estimation in simple terms with Calculati...Dr.Costas Sachpazis
Terzaghi's soil bearing capacity theory, developed by Karl Terzaghi, is a fundamental principle in geotechnical engineering used to determine the bearing capacity of shallow foundations. This theory provides a method to calculate the ultimate bearing capacity of soil, which is the maximum load per unit area that the soil can support without undergoing shear failure. The Calculation HTML Code included.
About
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
• Remote control: Parallel or serial interface.
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• Remote control system for accessing CCR and allied system over serial or TCP.
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Technical Specifications
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
Key Features
Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
• Remote control: Parallel or serial interface
• Compatible with MAFI CCR system
• Copatiable with IDM8000 CCR
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
Application
• Remote control: Parallel or serial interface.
• Compatible with MAFI CCR system.
• Compatible with IDM8000 CCR.
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
• Easy in configuration using DIP switches.
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Immunizing Image Classifiers Against Localized Adversary Attacksgerogepatton
This paper addresses the vulnerability of deep learning models, particularly convolutional neural networks
(CNN)s, to adversarial attacks and presents a proactive training technique designed to counter them. We
introduce a novel volumization algorithm, which transforms 2D images into 3D volumetric representations.
When combined with 3D convolution and deep curriculum learning optimization (CLO), itsignificantly improves
the immunity of models against localized universal attacks by up to 40%. We evaluate our proposed approach
using contemporary CNN architectures and the modified Canadian Institute for Advanced Research (CIFAR-10
and CIFAR-100) and ImageNet Large Scale Visual Recognition Challenge (ILSVRC12) datasets, showcasing
accuracy improvements over previous techniques. The results indicate that the combination of the volumetric
input and curriculum learning holds significant promise for mitigating adversarial attacks without necessitating
adversary training.
Welcome to WIPAC Monthly the magazine brought to you by the LinkedIn Group Water Industry Process Automation & Control.
In this month's edition, along with this month's industry news to celebrate the 13 years since the group was created we have articles including
A case study of the used of Advanced Process Control at the Wastewater Treatment works at Lleida in Spain
A look back on an article on smart wastewater networks in order to see how the industry has measured up in the interim around the adoption of Digital Transformation in the Water Industry.
Hybrid optimization of pumped hydro system and solar- Engr. Abdul-Azeez.pdffxintegritypublishin
Advancements in technology unveil a myriad of electrical and electronic breakthroughs geared towards efficiently harnessing limited resources to meet human energy demands. The optimization of hybrid solar PV panels and pumped hydro energy supply systems plays a pivotal role in utilizing natural resources effectively. This initiative not only benefits humanity but also fosters environmental sustainability. The study investigated the design optimization of these hybrid systems, focusing on understanding solar radiation patterns, identifying geographical influences on solar radiation, formulating a mathematical model for system optimization, and determining the optimal configuration of PV panels and pumped hydro storage. Through a comparative analysis approach and eight weeks of data collection, the study addressed key research questions related to solar radiation patterns and optimal system design. The findings highlighted regions with heightened solar radiation levels, showcasing substantial potential for power generation and emphasizing the system's efficiency. Optimizing system design significantly boosted power generation, promoted renewable energy utilization, and enhanced energy storage capacity. The study underscored the benefits of optimizing hybrid solar PV panels and pumped hydro energy supply systems for sustainable energy usage. Optimizing the design of solar PV panels and pumped hydro energy supply systems as examined across diverse climatic conditions in a developing country, not only enhances power generation but also improves the integration of renewable energy sources and boosts energy storage capacities, particularly beneficial for less economically prosperous regions. Additionally, the study provides valuable insights for advancing energy research in economically viable areas. Recommendations included conducting site-specific assessments, utilizing advanced modeling tools, implementing regular maintenance protocols, and enhancing communication among system components.
CFD Simulation of By-pass Flow in a HRSG module by R&R Consult.pptxR&R Consult
CFD analysis is incredibly effective at solving mysteries and improving the performance of complex systems!
Here's a great example: At a large natural gas-fired power plant, where they use waste heat to generate steam and energy, they were puzzled that their boiler wasn't producing as much steam as expected.
R&R and Tetra Engineering Group Inc. were asked to solve the issue with reduced steam production.
An inspection had shown that a significant amount of hot flue gas was bypassing the boiler tubes, where the heat was supposed to be transferred.
R&R Consult conducted a CFD analysis, which revealed that 6.3% of the flue gas was bypassing the boiler tubes without transferring heat. The analysis also showed that the flue gas was instead being directed along the sides of the boiler and between the modules that were supposed to capture the heat. This was the cause of the reduced performance.
Based on our results, Tetra Engineering installed covering plates to reduce the bypass flow. This improved the boiler's performance and increased electricity production.
It is always satisfying when we can help solve complex challenges like this. Do your systems also need a check-up or optimization? Give us a call!
Work done in cooperation with James Malloy and David Moelling from Tetra Engineering.
More examples of our work https://www.r-r-consult.dk/en/cases-en/
Student information management system project report ii.pdfKamal Acharya
Our project explains about the student management. This project mainly explains the various actions related to student details. This project shows some ease in adding, editing and deleting the student details. It also provides a less time consuming process for viewing, adding, editing and deleting the marks of the students.
Saudi Arabia stands as a titan in the global energy landscape, renowned for its abundant oil and gas resources. It's the largest exporter of petroleum and holds some of the world's most significant reserves. Let's delve into the top 10 oil and gas projects shaping Saudi Arabia's energy future in 2024.
Automobile Management System Project Report.pdfKamal Acharya
The proposed project is developed to manage the automobile in the automobile dealer company. The main module in this project is login, automobile management, customer management, sales, complaints and reports. The first module is the login. The automobile showroom owner should login to the project for usage. The username and password are verified and if it is correct, next form opens. If the username and password are not correct, it shows the error message.
When a customer search for a automobile, if the automobile is available, they will be taken to a page that shows the details of the automobile including automobile name, automobile ID, quantity, price etc. “Automobile Management System” is useful for maintaining automobiles, customers effectively and hence helps for establishing good relation between customer and automobile organization. It contains various customized modules for effectively maintaining automobiles and stock information accurately and safely.
When the automobile is sold to the customer, stock will be reduced automatically. When a new purchase is made, stock will be increased automatically. While selecting automobiles for sale, the proposed software will automatically check for total number of available stock of that particular item, if the total stock of that particular item is less than 5, software will notify the user to purchase the particular item.
Also when the user tries to sale items which are not in stock, the system will prompt the user that the stock is not enough. Customers of this system can search for a automobile; can purchase a automobile easily by selecting fast. On the other hand the stock of automobiles can be maintained perfectly by the automobile shop manager overcoming the drawbacks of existing system.
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Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the Fish, Labeo Rohita
1. International Journal of Pharmaceutical Science Invention
ISSN (Online): 2319 – 6718, ISSN (Print): 2319 – 670X
www.ijpsi.org Volume 5 Issue 4 ‖ July 2016 ‖ PP. 47-51
www.ijpsi.org 47 | P a g e
Evaluation of LdhIsozymes Following the Treatment of Methyl
Parathion in the Fish, Labeo Rohita
*S. N. C. Ray and *1
R. C. Sinha
*Centre for Environment and Nature Conservation, Department of Zoology, Patna University, Patna-800005,
India
1
Chief Executive, Centre for Environment and Nature Conservation, Department of Zoology, Patna University,
Patna- 800005, India
ABSTRACT
Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate
metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions.
The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of
methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of
methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish
Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of
human serum suggesting liver damage. This was also confirmed by histopathological studies.
Keywords: Lactate dehydrogenase, isozymes, methyl parathion, Labeo rohita.
I. Introduction
Methyl parathion, an organophosphate pesticide, extensively used in agriculture is known to be responsible for a
number of physiological and biochemical disturbances to the non-target organisms. It is known to be a
neurotoxic and hepatotoxic pesticide. It has been reported that some of the serum enzymes are useful in
diagnosing liver diseases [1, 2]. Cellular enzymes in the extracellular space, although of no further metabolic
function in this space are still of benefit because they serve as indicators suggestive of disturbances of cellular
integrity induced by foreign interferences. Lactatedehydrogenase (LDH) is a cytoplasmic enzyme present in
essentially all major organ systems. The extracellular appearance of LDH is used to detect cell damage or cell
death [3]. Due to its extraordinarily interspersed distribution in the body, serum LDH is abnormal in many
disorders [4]. It is released into peripheral space after cell rupture or cell death caused by e.g. excess heat or
cold, starvation, exposure to pesticides or toxins, therefore the total serum LDH is highly sensitive but non-
specific test.
In order to optimize the diagnostic value, LDH isozymes can be measured [5]. Lactate dehydrogenase (EC
3.1.1.27) is one of the chief enzymes of carbohydrate metabolism which catalyzes the oxidation of lactate and
reduction of pyruvate anaerobic glycolysis. It is a dimeric molecule consisting of two separate loci which code
for A and B sub-units of this enzyme. The A and B subunits indiscriminately associate and form five tetrameric
isozymes (A4, A3B1, A2B2, A3, B1 and B4) [6]. Isozymes are multiple forms of single enzyme which have
different isoelectric points and therefore be separated by electrophoresis. Electrophoretic separation of LDH
were done extensively on different tissues of various animals from which it revealed that the enzyme exhibited
multimolecular forms and functions [7].
Classically, five LDH isozymes are present in the serum, of which the electrophoretically slowest moving band
(LDH5) corresponding to the liver isozymes. The appearance of LDH5 isozyme in the serum indicates
hepatocellular damage. Since the LDH isozymes patterns are different in various organs and tissues, its serum
analysis may be useful in establishing the cell injury.
LDH in fish has always been the subject of much attention. But not much information is available with the
changes in the expression of LDH on the toxicity of methyl parathion. The aim of the present paper is to
evaluate the expression of LDH isozymes in the serum of fish, Labeo rohita after exposure to sub- lethal
concentration of methyl parathion for 96h using a fully automatic clinicalelectrophoresis and densitometer.
II. Materials And Methods
Ethical Statement: Presently, we do not have any Ethical Committee in our University. But however, we have
followed the ethical norms, which are being followed elsewhere which is evident in the Materials & Methods
Section.
Maintenance of animals:Labeo rohita, a common carp was obtained from the local hatchery. Fishes were
acclimated to laboratory conditions for about 5-7 days. They were kept in aquarium tank (250 L) and water was
2. Evaluation Of LdhIsozymes Following The Treatment Of Methyl Parathion In The Fish...
www.ijpsi.org 48 | P a g e
constantly aerated by a static system. During the acclimation period, they were given artificial (commercial)
feed composed of ground shrimps available in the local market to avoid the possible effects of starvation. The
feeding and maintenance of the fishes and physico-chemical characteristics of the aquaria water were measured
(Tab. 1). Short-term test of acute toxicity over a period of 96h were performed on the fishes following the
renewal of bioassay. The fishes were exposed intra-coelomatically with 1/3rd
of LC50 value of the pesticide
methyl parathion. After 24, 48, 72 and 96 h of exposure fishes were processed for further investigations.
Determination of LC50: The experiments were repeated several times and only arithmetic mean of the
experiments at each concentration was taken to express the results. LC50 values were determined by EPA Probit
analysis program [8]. The LC50 of methyl parathion for the fish Labeo rohita was 16.8 ppm.
Blood collection:The fishes were taken out of the aquarium water individually through fish net with a minimum
possible disturbance. After preliminary investigations, the blood samples were collected from caudal fin as
described by many authors. In the present study, the blood collection from caudal fin had to be abandoned
because there was an unusual elevation in Lactate dehydrogenase (LDH) and Creatine phosphokinase (CPK)
activities which were recorded due to leakage from the surrounding muscle tissues. Thus, cardiac sampling was
the only suitable method available as an alternative to obtain blood under the present study. After the blood
collection, the serum were separated and processed for enzymatic assays.
Histological procedures: Sample of fish from both control and experimental groups were excised, rinsed in
physiological saline fixed in Bouin’s fluid and thereafter processed for microtomy. The tissues were dehydrated
in ethyl alcohol with series of ascending concentrations, embedded in paraffin and sectioned, which were
subsequently stained with haemotoxylin and eosin. The slides were examined under low power (4X) objective
and high power (10X) objective with binocular microscope. Photomicrographs were taken at high power (100X)
zoom lens digital camera with software DigiPro 5.0 attached in the microscopeLabomed, model- LX400 and
downloaded into a computer.
Separation of LDH isozymes by Fully Automatic Clinical Electrophoretic Unit:Fully automatic clinical
electrophoretic unit is a system software controlled electrophoretic separation having a voltage, current and run
time control, operated by an inbuilt programme present in the driver software of the electrophoresis. Fully
Automatic Clinical Electriphoresis Model- Interlab- Pretty was used for the present study having software
namely ‘Interlab’. The system constituted of an automatic dispensing system for samples, a Teflon coated
Peltier Plate for placing previously casted agarose gel over sterile plastic plate having bar- code over it and
valve- compressor system for pouring appropriate amount of staining, destaining and washing solutions over the
gel plate after completion of the migration. About 30µl of previously separated non-hemolyzed serum were
placed in the sample space and loaded over the gel plate by the automatic dispensing system to achieve single
straight line of application. The temperature for migration was set at 290
C, time for migration was 5 minutes,
voltage for migration was 400V and after migration secondary treatment of the gel was needed which was done
in another chamber with an incubation period of 30 minutes at 450
C after addition of staining solution. The
staining solution contained pyruvate as substrate and NitroblueTetrazolium (NBT), NBT was reduced to
formazan by NADH, also present in the staining solution. After the treatment with the staining solution, which
stained the LDH isozyme bands, fixation of the band was achieved in the above mentioned external chamber.
After staining and fixing the bands, the gel plate was re-placed into the electrophoretic chamber for repeated
destaining with destaining solution comprising of acetic acid of strength 5% (v/v). Then washing was done with
a surfactant solution for a number of times previously fed in the driver software. After completion of these steps,
scanning of the gel plate was done in the densitometer with adriver software ‘Scanlab’ provided by the
manufacturer.
III. Results And Discussion
Table-I. Physico-chemical characteristics of aquaria water
Sl no. Parameter Value
1. Temperature (24±2)0
C
2. pH 7.1± 0.2 at 240
C
3. Dissolved Oxygen 8.5 ±0.5 mg/L
4. Total Hardness 23.4± 3.4mg CaCO3/L
5. Conductivity <10 µs/cm
Table-II. Name of LDH isozyme bands with their percentages in the electrophorogram from cathode to anode
Name of the LDH isozyme
band (from Cathode to Anode)
Control 24h 48h 72h 96h
A 55.6% 65.6% 52.1% 45.7% 41.8%
A’ --- --- --- --- ---
B 31.3% 34.4% 47.9% 54.3% 42.2%
B’ 13.1% --- --- --- 16.0%
3. Evaluation Of LdhIsozymes Following The Treatment Of Methyl Parathion In The Fish...
www.ijpsi.org 49 | P a g e
Figure 1. Agarose Gel Plate with positioning of the LDH isozyme bands of human and fishLabeo rohita
(Control and treated 24h, 48h, 72h and 96h)
List of Abbreviations: HC= Hepatic Cord; CV= Central Vein; S= Sinusoid; BP= Bile Plug; SP= Sinusoidal
Space; FN= Fragmented Nucleus
Plate 1: Control liver of L. Rohita
Plate 2: 24h Methyl parathion treated Plate 3: 48h Methyl parathion treated
liver of L. Rohita liver of L. Rohita
4. Evaluation Of LdhIsozymes Following The Treatment Of Methyl Parathion In The Fish...
www.ijpsi.org 50 | P a g e
Plate 4: 72h Methyl parathion treated Plate 5: 96h Methyl parathion treated
liver of L. Rohita liver of L. Rohita
Figure 2: Plate numbers showing the Liver histopathology of the fish Labeo rohita
It is well known that LDH is present in the cells of almost every body tissue and in the case of damage it is
released into the blood [9, 10]. Analyses have revealed LDH to be suitable biomarker since the results obtained
after expositions of animals to different toxicants were found to be significant. It has already been mentioned
earlier that LDH is a highly sensitive but not specific biomarker. Earlier, the authors have reported that LDH in
the serum of the fish Cirrhinus mrigala increased significantly following the treatment of sub-lethal dose of
methyl parathion [11]. Thus, in the present investigation, supplementary information is being reported by
electrophoretic separation and quantification of LDH isozymes in the fish Labeo rohita as a function of methyl
parathion. The LDH isozymeelectrophorogram have shown that there are four bands from cathode to anode. The
bands of electrophorogram of LDH isozymes are designated as LDH A, LDH A’, LDH B and LDH B’. In the
control fish, LDH A is 55.6%, LDH A’ is absent whereas LDH B is 31.3% and LDH B’ is 13.1% (Table-2). But
following the treatment of methyl parathion LDH A increased to 65.6% during 24h and thereafter decreased to
52.1%, 45.7% and 41.8% during 48h, 72h and 96h respectively. Whereas, LDH B increased to 34.4% in 24h
and further increased to 47.9% and 54.3% in 48h and 72h respectively but decreased to 42.2% during 96h. LDH
B’ was absent during 24, 48 and 72h but present during 96h.
The differential percentage of increase or decrease of different isozymes in the treated fish during different
periods may be due to the differential mode of action of methyl parathion in different tissues. Secondly, gene
duplication as well as post- translational processing has been proposed as important factors in modulating tissue
specific enzymes [12]. LDH A’ is missing in the control fish as well as in the treated fish. Similarly, B’ is absent
following the treatment of methyl parathion during 24h, 48h, 72h but present in 96h (Table-2). The increase/
decrease/ absence of some of the isozymes could be interpreted as a result of certain mutational events that
would have occurred in the regulatory genes which could leadto inhibition, alteration or constitutive gene
expression. This is the reason some bands of isozyme became more intense or faint (Fig. 1). The differences of
the distribution of the different LDH isozymesin the serum reflect differences in their metabolic activity.
It is interesting to observe that three distinct bands appear in the 96h (Fig. 1, Lane- 3) the third cathodic band,
the slowest moving band which is almost parallel to LDH5 of human suggesting liver damage. Histological
examination of the tissues could serve as complementary evidence [13] to enzyme studies towards revealing any
distortion/damage to the normal structure of the tissues [14, 15, 16]. In the present study we have observed the
histological changes in the liver of the fish, Labeo rohita as a function of methyl parathion.The histological
changes in the animal tissues provide a rapid, cost effective method to detect effect of toxicants.
The liver of the control fish comprise of a continuous mass of large hexagonal hepatic cells containing more or
less spherical nucleus. They are located among sinusoids and forming cord like structures known as hepatic cell
cord. In fish these structures are generally obscure. Bile canaliculus is centrally located in each cord. There is no
clear cut division of hepatic cells with lobules. These cells contain granular cytoplasm with distinct nuclei either
exocentric or slightly centrally placed. Hepatic cells have many vital functions other than secretion of bile. They
play an important role in protein, lipid and carbohydrate metabolism. Detoxification is another important
function (Plate 1).
After the treatment of 1/3rd
of LC50 of methyl parathion, following changes were observed in 24h, 48h, 72h and
96h.
In 24h, hepatic cords were disorganized; bile pigment appeared; some hepatic cell nucleus divided; central vein
became clear; interstitial space was filled with hazy liquid, very few RBCs were in between the hepatic cells;
hepatic cells became dense in appearance (Plate- 2).
5. Evaluation Of LdhIsozymes Following The Treatment Of Methyl Parathion In The Fish...
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In 48h, the spaces between the hepatic cords were comparatively more; hepatic cord became disorganized;
central vein became disorganized with more RBCs (Plate-3).
In 72h, central vein ruptured; lack of rounded structure of hepatic nuclei; hepatic cell membrane was found to be
splitting leading to necrotic changes; formation of sinusoidal space was more nuclear disintegration was very
clear and found scattered in the sinusoid space; appearance of bile pigment(Plate-4).
In 96h, sinusoidal space was more while the hepatic cells joined each other; nucleus was rounded; central vein
had less RBCs (Plate-5).
The fish liver histology is an indicator of chemical toxicity and very useful method to study the effects of
exposure of aquatic animals to toxicants present in the aquatic system [17]. The liver is the primary organ for
detoxification of the xenobiotics. As mentioned above, the histopathological findings highly corroborates with
the biochemical results i.e. during 72h to 96h the hepatic cell membrane ruptures with the release of isozymes
especially LDH5 (Fig.- 1, lane-3) clearly suggesting that the LDH5 isozyme is the indicative of liver damage.
Thus, it is concluded that histopathological changes coupled with biochemical changes could serve as valuable
biomarkers for methyl parathion exposed fish.
Acknowledgement
The authors are grateful to DBT, Govt. of India for financial assistance for the project (Sanction Letter no.
BT/PR/4762/BCE/8/903/2012; Dated: 11/3/2013). The authors also thank the Head of the Department of
Zoology for extending all logistic support.
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