Enhanced Drug Uptake on Application of Electroporation in a Single-Cell Model.

Electroporation method is a useful tool for delivering drugs into various diseased tissues in the human body. As a result of an applied electric field, drug particles enter the intracellular compartment through the temporarily permeabilized cell membrane. Consequently, electroporation method allows better penetration of the drug into the diseased tissue and improves treatment clinically. In this study, a more generalized model of drug transport in a single cell is proposed. The model is able to capture non-homogeneous drug transport in the cell due to non-uniform cell membrane permeabilization. Several numerical experiments are conducted to understand the effects of electric field and drug permeability on drug uptake into the cell. Through investigation, the appropriate electric field and drug permeability are identified, which lead to sufficient drug uptake into the cell. This model can be used by experimentalists to get information prior to conduct any experiment, and it may help reduce the number of actual experiments that might be conducted otherwise.

Rete middle cerebral artery presenting with a ruptured aneurysm.

The developmental anomaly of the middle cerebral artery (MCA) is a rare occurrence. Rete MCA, also known as "unfused," "twig-like," and aplastic MCA, is an even rarer anomaly. These immature rete vessels are prone to flow-related aneurysms which rupture themselves presenting with intraparenchymal hematoma. We here with present a case of rete MCA with a ruptured aneurysm and discuss its pathophysiology and management.

Supramolecular associations between atypical oxidative phosphorylation complexes of Euglena gracilis.

In vivo associations of respiratory complexes forming higher supramolecular structures are generally accepted nowadays. Supercomplexes (SC) built by complexes I, III and IV and the so-called respirasome (I/III2/IV) have been described in mitochondria from several model organisms (yeasts, mammals and green plants), but information is scarce in other lineages. Here we studied the supramolecular associations between the complexes I, III, IV and V from the secondary photosynthetic flagellate Euglena gracilis with an approach that involves the extraction with several mild detergents followed by native electrophoresis. Despite the presence of atypical subunit composition and additional structural domains described in Euglena complexes I, IV and V, canonical associations into III2/IV, III2/IV2 SCs and I/III2/IV respirasome were observed together with two oligomeric forms of the ATP synthase (V2 and V4). Among them, III2/IV SC could be observed by electron microscopy. The respirasome was further purified by two-step liquid chromatography and showed in-vitro oxygen consumption independent of the addition of external cytochrome c.

Urine Extracellular Vesicle GATA2 mRNA Discriminates Biopsy Result in Men with Suspicion of Prostate Cancer.

PURPOSE: Prostate specific antigen has limited performance in detecting prostate cancer. The transcription factor GATA2 is expressed in aggressive prostate cancer. We analyzed the predictive value of urine extracellular vesicle GATA2 mRNA alone and in combination with a multigene panel to improve detection of prostate cancer and high risk disease. MATERIALS AND METHODS: GATA2 mRNA was analyzed in matched extracellular vesicles isolated from urines before and after prostatectomy (16) and paired urine and tissue prostatectomy samples (19). Extracellular vesicle GATA2 mRNA performance to distinguish prostate cancer and high grade disease was tested in training (52) and validation (165) cohorts. The predictive value of a multigene score including GATA2, PCA3 and TMPRSS2-ERG (GAPT-E) was tested in both cohorts. RESULTS: Confirming its prostate origin, urine extracellular vesicle GATA2 mRNA levels decreased significantly after prostatectomy and correlated with prostate cancer tissue GATA2 mRNA levels. In the training and validation cohort GATA2 discriminated prostate cancer (AUC 0.74 and 0.66) and high grade disease (AUC 0.78 and 0.65), respectively. Notably, the GAPT-E score improved discrimination of prostate cancer (AUC 0.84 and 0.72) and high grade cancer (AUC 0.85 and 0.71) in both cohorts when compared with each biomarker alone and PT-E (PCA3 and TMPRSS2-ERG). A GAPT-E score for high grade prostate cancer would avoid 92.1% of unnecessary prostate biopsies, compared to 61.9% when a PT-E score is used. CONCLUSIONS: Urine extracellular vesicle GATA2 mRNA analysis improves the detection of high risk prostate cancer and may reduce the number of unnecessary biopsies.

Co-evolution of spliceosomal disassembly interologs: crowning J-protein component with moonlighting RNA-binding activity.

Spliceosome disassembly is catalyzed by the NineTeen-related (NTR) complex, which is constituted by several proteins, including Cwc23, Ntr1, and Ppr43. Cwc23 is an essential J-protein in Saccharomyces cerevisiae that recruits Ntr1, an NTC-related G-patch protein, to the spliceosome. Ntr1 interacts with Prp43, a DExD/H box RNA helicase protein, which facilitates the disassembly of spliceosomal intermediates. The interaction between Ntr1 and Prp43 is conserved and crucial for the disassembly process. However, the J-protein component of this complex is not studied in other eukaryotes. In silico analysis supported by results of yeast complementation and two-hybrid studies suggests that while Prp43 is highly conserved, both Ntr1 and Cwc23 are co-evolving components of the disassembly triad. The J-domain of Cwc23, which is otherwise dispensable for its function, is highly conserved, whereas the functionally critical C-terminus has significantly diverged in Cwc23 orthologs. Some eukaryotic orthologs of Cwc23 contain a distinct RNA recognition motif at their C-terminus and are able to bind RNA in vitro. Based on the results presented in this study, we propose that RNA-binding activity in some eukaryotic orthologs of Cwc23 might provide additional functional diversity or robustness to the J-protein/Hsp70 machine in spliceosomal remodelling processes.

An alkali tolerant α-l-rhamnosidase from Fusarium moniliforme MTCC-2088 used in de-rhamnosylation of natural glycosides.

Analkali tolerant α-l-rhamnosidase has been purified to homogeneity from the culture filtrate of a new fungal strain, Fusarium moniliforme MTCC-2088, using concentration by ultrafiltration and cation exchange chromatography on CM cellulose column. The molecular mass of the purified enzyme has been found to be 36.0 kDa using SDS-PAGE analysis. The Km value using p-nitrophenyl-α-l-rhamnopyranoside as the variable substrate in 0.2 M sodium phosphate buffer pH10.5 at50 °C was 0.50 mM. The catalytic rate constant was15.6 s-1giving the values of kcat/Km is 3.12 × 104M-1 s-1. The pH and temperature optima of the enzyme were 10.5 and 50 °C, respectively. The purified enzyme had better stability at 10 °C in basic pH medium. The enzyme derhamnosylated natural glycosides like naringin to prunin, rutin to isoquercitrin and hesperidin to hesperetin glucoside. The purified α-l-rhamnosidase has potential for enhancement of wine aroma.

Cost-effective way to reduce stimulant-abuse among gay/bisexual men and transgender women: a randomized clinical trial with a cost comparison.

OBJECTIVES: A randomized controlled study was conducted with 422 homeless, stimulant-using gay/bisexual (G/B) men and 29 transgender women (n = 451) to assess two community-based interventions to reduce substance abuse and improve health: (a) a nurse case-managed program combined with contingency management (NCM + CM) versus (b) standard education plus contingency management (SE + CM). STUDY DESIGN: Hypotheses tested included: a) completion of hepatitis A/B vaccination series; b) reduction in stimulant use; and c) reduction in number of sexual partners. METHODS: A deconstructive cost analysis approach was utilized to capture direct costs associated with the delivery of both interventions. Based on an analysis of activity logs and staff interviews, specific activities and the time required to complete each were analyzed as follows: a) NCM + CM only; b) SE + CM only; c) time to administer/record vaccines; and d) time to receive and record CM visits. Cost comparison of the interventions included only staffing costs and direct cash expenditures. RESULTS: The study outcomes showed significant over time reductions in all measures of drug use and multiple sex partners, compared to baseline, although no significant between-group differences were detected. Cost analysis favored the simpler SE + CM intervention over the more labor-intensive NCM + CM approach. Because of the high levels of staffing required for the NCM relative to SE, costs associated with it were significantly higher. CONCLUSIONS: Findings suggest that while both intervention strategies were equally effective in achieving desired health outcomes, the brief SE + CM appeared less expensive to deliver.

Interaction between the photoprotective protein LHCSR3 and C2S2 Photosystem II supercomplex in Chlamydomonas reinhardtii.

Photosynthetic organisms can thermally dissipate excess of absorbed energy in high-light conditions in a process known as non-photochemical quenching (NPQ). In the green alga Chlamydomonas reinhardtii this process depends on the presence of the light-harvesting protein LHCSR3, which is only expressed in high light. LHCSR3 has been shown to act as a quencher when associated with the Photosystem II supercomplex and to respond to pH changes, but the mechanism of quenching has not been elucidated yet. In this work we have studied the interaction between LHCSR3 and Photosystem II C2S2 supercomplexes by single particle electron microscopy. It was found that LHCSR3 predominantly binds at three different positions and that the CP26 subunit and the LHCII trimer of C2S2 supercomplexes are involved in binding, while we could not find evidences for a direct association of LHCSR3 with the PSII core. At all three locations LHCSR3 is present almost exclusively as a dimer.

Supercomplexes of plant photosystem I with cytochrome b6f, light-harvesting complex II and NDH.

Photosystem I (PSI) is a pigment-protein complex required for the light-dependent reactions of photosynthesis and participates in light-harvesting and redox-driven chloroplast metabolism. Assembly of PSI into supercomplexes with light harvesting complex (LHC) II, cytochrome b6f (Cytb6f) or NAD(P)H dehydrogenase complex (NDH) has been proposed as a means for regulating photosynthesis. However, structural details about the binding positions in plant PSI are lacking. We analyzed large data sets of electron microscopy single particle projections of supercomplexes obtained from the stroma membrane of Arabidopsis thaliana. By single particle analysis, we established the binding position of Cytb6f at the antenna side of PSI. The rectangular-shaped Cytb6f dimer binds at the side where Lhca1 is located. The complex binds with its short side rather than its long side to PSI, which may explain why these supercomplexes are difficult to purify and easily disrupted. Refined analysis of the interaction between PSI and the NDH complex indicates that in total up to 6 copies of PSI can arrange with one NDH complex. Most PSI-NDH supercomplexes appeared to have 1-3 PSI copies associated. Finally, the PSI-LHCII supercomplex was found to bind an additional LHCII trimer at two positions on the LHCI side in Arabidopsis. The organization of PSI, either in a complex with NDH or with Cytb6f, may improve regulation of electron transport by the control of binding partners and distances in small domains.

Atypical composition and structure of the mitochondrial dimeric ATP synthase from Euglena gracilis.

Mitochondrial respiratory-chain complexes from Euglenozoa comprise classical subunits described in other eukaryotes (i.e. mammals and fungi) and subunits that are restricted to Euglenozoa (e.g. Euglena gracilis and Trypanosoma brucei). Here we studied the mitochondrial F1FO-ATP synthase (or Complex V) from the photosynthetic eukaryote E. gracilis in detail. The enzyme was purified by a two-step chromatographic procedure and its subunit composition was resolved by a three-dimensional gel electrophoresis (BN/SDS/SDS). Twenty-two different subunits were identified by mass-spectrometry analyses among which the canonical α, ß, γ, δ, ε, and OSCP subunits, and at least seven subunits previously found in Trypanosoma. The ADP/ATP carrier was also associated to the ATP synthase into a dimeric ATP synthasome. Single-particle analysis by transmission electron microscopy of the dimeric ATP synthase indicated that the structures of both the catalytic and central rotor parts are conserved while other structural features are original. These new features include a large membrane-spanning region joining the monomers, an external peripheral stalk and a structure that goes through the membrane and reaches the inter membrane space below the c-ring, the latter having not been reported for any mitochondrial F-ATPase.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors: Present perspectives and future horizons.

AIMS: Our comprehensive review highlights the drug development and pharmacogenomics leading to the recent approval of PCSK9 inhibitors. We also review the anticipated future advances into the uses of PCSK9 inhibition. BACKGROUND: Despite the present advances in pharmacotherapy, atherosclerotic cardiovascular disease (ASCVD) remains the leading cause of mortality worldwide. Low density lipoprotein-cholesterol (LDL-C) lowering is the primary target for ASCVD risk reduction, showing demonstrable benefits in mortality. However, 70% of events occur even in the presence of statins. This residual risk may be approached with additional LDL-C reduction. Statin intolerance is a common clinical concern affecting adherence and the benefit with statins. There is also significant variation of individual lipid-lowering. Following rapid development, proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors have progressed from genetic observations, to mechanistic studies, to closer realization of the goal of CVD risk reduction. This review discusses the science behind PCSK9 inhibition, evidence of trials involving efficacy and safety, and reflections of its present and future role in clinical care, especially in high-risk patients with ASCVD, persons with suboptimal responses to statins and familial hyperlipidemia. Monoclonal antibodies have demonstrated LDL-C lowering of up to 57% as monotherapy and up to 73% when added to statins. Statins have limited efficacy in reduction of LDL-C due to an increased number of LDL-receptors. Elevated lipoprotein (a) levels may also be significantly lowered by PCSK9i. The journey from discovery to PSCK9 target validation took less than five years, and development and approval of therapeutic modalities for PCSK9 inhibitors happened over the next seven. This review highlights the drug development and pharmacogenomics leading to the recent approval of two agents, alirocumab and evolocumab, with a third bococizumab, and other novel approaches to the pathway pending. DATA SYNTHESIS: We searched MEDLINE database via Pubmed for reviews, research publications and relevant trials available on PCSK9 inhibition. CONCLUSION: Despite decades of medical advances, ASCVD remains one of the major causes of morbidity and mortality worldwide. Statin use has multiplied since the validation of LDL hypothesis, however, it is undeniable a more effective and well-tolerated agent is needed in significant number or patients. With the arrival of the era of unprecedented CV protection with PCSK9 inhibition, this exciting new therapy holds a pivotal promise as the future of lipid management. The data available already indicate safety, tolerability and superb efficacy of these agents, which are already changing contemporary cholesterol management. The rapid translation of innovative basic science research into drug development may lead to CV outcomes reduction and confirm that this pathway will become prominently utilized.

Consequences of state transitions on the structural and functional organization of photosystem I in the green alga Chlamydomonas reinhardtii.

State transitions represent a photoacclimation process that regulates the light-driven photosynthetic reactions in response to changes in light quality/quantity. It balances the excitation between photosystem I (PSI) and II (PSII) by shuttling LHCII, the main light-harvesting complex of green algae and plants, between them. This process is particularly important in Chlamydomonas reinhardtii in which it is suggested to induce a large reorganization in the thylakoid membrane. Phosphorylation has been shown to be necessary for state transitions and the LHCII kinase has been identified. However, the consequences of state transitions on the structural organization and the functionality of the photosystems have not yet been elucidated. This situation is mainly because the purification of the supercomplexes has proved to be particularly difficult, thus preventing structural and functional studies. Here, we have purified and analysed PSI and PSII supercomplexes of C. reinhardtii in states 1 and 2, and have studied them using biochemical, spectroscopic and structural methods. It is shown that PSI in state 2 is able to bind two LHCII trimers that contain all four LHCII types, and one monomer, most likely CP29, in addition to its nine Lhcas. This structure is the largest PSI complex ever observed, having an antenna size of 340 Chls/P700. Moreover, all PSI-bound Lhcs are efficient in transferring energy to PSI. A projection map at 20 Å resolution reveals the structural organization of the complex. Surprisingly, only LHCII type I, II and IV are phosphorylated when associated with PSI, while LHCII type III and CP29 are not, but CP29 is phosphorylated when associated with PSII in state2.

Catchment delineation and morphometric analysis using geographical information system.

The geographical information system (GIS) has emerged as an efficient tool in delineation of drainage patterns of watershed planning and management. The morphometric parameters of basins can address linear, areal and relief aspects. The study deals with the integrated watershed management of Baliya micro-watersheds, located in the Udaipur district of Rajasthan, India. Morphometric analysis in hydrological investigation is an important aspect and it is inevitable in the development and management of drainage basins. The determination of linear, areal and relief parameters indicate fairly good significance. The low value of the bifurcation ratio of 4.19 revealed that the drainage pattern has not been distorted by structural disturbance. The high value of the elongation ratio (0.68) compared to the circulatory ratio (0.27) indicates an elongated shape of the watershed. The high value of drainage density (5.39 km/km(2)) and stream frequency (12.32) shows that the region has impermeable subsoil material under poor vegetative cover with a low relief factor. The morphometric parameters of relief ratio (0.041) and relative relief (0.99%) show that the watershed can be treated using GIS techniques to determine the morphometric presence of dendritic drainage pattern, with a view to selecting the soil and water conservation measures and water harvesting.

Comparing the Effectiveness of Traditional Periodontal Surgery Versus Non-Surgical Therapy: A Randomized Controlled Trial.

Background: Periodontal disease, characterized by inflammation and infection of the supporting structures of teeth, poses a significant oral health challenge. Traditional periodontal surgery and non-surgical therapy, such as scaling and root planing, are established treatment approaches for addressing periodontal disease. Materials and Methods: The study enrolled 120 adult patients diagnosed with moderate to severe periodontal disease. Participants were randomly allocated to one of two groups: the traditional surgery group (TSG) or the non-surgical therapy group (NSTG). In the TSG, patients underwent traditional periodontal surgery, which included flap surgery and grafts when deemed necessary. The surgical procedures were performed by experienced periodontal surgeons. In contrast, the NSTG received non-surgical therapy in the form of scaling and root planing administered by trained dental hygienists. Outcome measures encompassed clinical parameters and patient-centered outcomes. Periodontal pocket depth and clinical attachment level, both measured in millimeters, were assessed at baseline, 3 months, and 6 months. Patient-reported outcomes, including pain, discomfort, and satisfaction, were collected through standardized questionnaires at each follow-up visit. Results: Patients in the TSG experienced a notable reduction in pocket depth from a baseline of 6.8 mm to 3.7 mm at the 6-month mark, resulting in a change of -3.1 mm. Conversely, the NSTG exhibited a reduction from 6.7 mm to 4.0 mm, with a change of -2.7 mm. In the TSG, the baseline attachment level of 7.2 mm decreased to 5.1 mm at 6 months, indicating a change of -2.1 mm. In the NSTG, the attachment level decreased from 7.1 mm to 5.5 mm, resulting in a change of -1.6 mm. Patients in the TSG reported an average pain score of 3.6 on a 1-10 scale, discomfort of 4.2, and satisfaction of 7.8. In contrast, patients in the NSTG reported lower pain (2.1) and discomfort (2.9) scores but similar satisfaction levels (8.4). Conclusion: In this randomized controlled trial (RCT), both traditional periodontal surgery and non-surgical therapy demonstrated improvements in clinical parameters and patient-reported outcomes. Traditional surgery resulted in greater reductions in periodontal pocket depth and clinical attachment loss at the 6-month follow-up.

Prioritizing areas for malaria control using geographical information system in Sonitpur district, Assam, India.

OBJECTIVE: To identify the malaria hot spots at health subcentre level in an endemic district using a geographical information system (GIS). The results will be useful for rapid retrieval of malaria information, and to prioritize malaria control efforts in identified hot spots. STUDY DESIGN: Extraction, analysis and synthesis of relevant data. METHODS: Malaria epidemiological data from 2006 to 2009 were analysed to determine the annual parasitic index, slide positivity rate, annual blood examination rate and Plasmodium falciparum percentage for each health subcentre in the district. Maps were produced using GIS, and integrated to identify the malaria hotspots. RESULTS: Out of 288 health subcentres, GIS identified 10 hot spots at extremely high risk of malaria and 14 hot spots at high risk of malaria. Malaria may flare up in these hot spots whenever favourable transmission conditions arise. Health authorities have been advised to establish control measures in these selected hot spots for timely prevention. CONCLUSION: There is a need for adequate monitoring and allocation of available resources for better interventions in the malaria hotspots. The GIS model used in this study can be used, even at village or cluster level, to pin point the malaria hot spots, and information can be updated and retrieved easily.

Optimization of virtual water flows in agriculture by changing cropping patterns using an integrated approach.

Utilizing available water resources efficiently is crucial to address both our present and future requirements and plays a vital role in safeguarding food security. This current investigation deals with assessment and optimizing water footprint (WF) and virtual water flow (VWF) for primary crops in Banas River Basin (BRB) using AquaCrop model with local datasets and district-level estimates. VWF in the basins were estimated by multiplying the WF of crops with the amount exported/imported, which is determined based on the difference between production and consumption in the basin. The possibility of changing the cropping patterns was evaluated for the potential reduction of the blue WF. Annual WF from primary crops in the basin amounts to 19,255 MCM/yr (70 % green, 21 % blue and 10 % grey WF, respectively). Banas basin is a net exporter of agriculture commodities with nearly 7391 MCM/yr of water flowing out of the basin due to agricultural exports of which approximately 265 MCM/yr is virtual blue water outflow. Crops having low economic water productivity of blue water are being grown in vast areas resulting in a high blue WF. The optimizing the cropping pattern can result in a 5-42 % lower blue water footprint with 11-39 % higher economic output under different scenarios with and without considering the consumption needs. Changing the cropping pattern and making trade plan to optimize the crop import/exports can be viable option for tackling the blue water scarcity issues in the basin. WF can be managed sustainably by improving water resource allocation for better economic, social, and environmental productivity and going for less aggressive agricultural production.

Evaluation of the Impact of Chlorhexidine Mouth Rinse on the Bond Strength of Polycarbonate Orthodontic Brackets: A Case-Control Study.

BACKGROUND: Shear bond analysis is the procedure used most frequently to gauge the tensile strength of adhesives incorporated in orthodontic treatments. In shear tensile strength analysis, pressure is placed as close as feasible to the interface between the orthodontic bracket and the surface of the tooth, parallel to the long axis of the tooth. Although numerous research on extracted teeth of human and bovine teeth have been conducted, there may still be variables such as pH, humidity, temperature, and others that could affect how these materials behave in the mouth cavity. The impact of chlorhexidine (CHX) on the binding capacity for non-metallic orthodontic brackets in vivo is not well understood. OBJECTIVE: The goal of the current study is to determine how mouth rinses containing 0.12% CHX affect the adhesive strength of polycarbonate orthodontic brackets. METHODS AND MATERIALS: Thirty-four patients were part of the test category, and they were instructed to wash their oral cavity for approximately 30 seconds using 20 ml of 0.12% CHX gluconate (Septodent). Thirty-four patients made up the control category and were instructed to wash their oral cavity for 30 seconds with a placebo mouthwash of a similar hue (20 ml). Both types of mouthwash were administered to the participants by an administrator who was not specifically involved in the trial and were kept in 120 ml labeled plastic bottles. The study participants were also kept unaware of the type of mouthwash. For the mouthwash utilized by study participants, a double-blinding technique was applied. RESULTS: Thirty-four patients were evaluated in the test category. Since the orthodontic bracket broke in two patients, therefore, 32 patients were evaluated in the control category. The mean value of the strength of the shear bond in the experimental category was 15.32 megapascal (Mpa). The SD value was 2.51. The mean value of the strength of the shear bond in the control subgroup was 15.63. On statistical analysis, the t-value was 0.47. The p-value was 0.671. The difference in findings of the strength of the shear bond was statistically non-significant. CONCLUSION: The results of this investigation allow us to draw the conclusion that the shear bond properties of polycarbonate orthodontic brackets are unaffected when treated with 0.12% CHX preceding the binding. The clinically meaningful adhesion strength was likewise attained by the polycarbonate orthodontic brackets.

Effect of novel Matrix Rhythm Therapy (MaRhyThe®) on neuropathic pain and maximum plantar pressure distribution among type 2 diabetes mellitus patients with peripheral neuropathy.

Background: There is a need for a non-pharmacological approach to reduce pain and plantar pressure in diabetic peripheral neuropathy (DPN). Matrix Rhythm Therapy (MaRhyThe®) is a therapeutic modality that works on the principle of physiologic rhythmic oscillations of the body cells. This study aimed to evaluate the effect of MaRhyThe® on neuropathic pain and maximum plantar pressure distribution among type 2 diabetes mellitus patients with peripheral neuropathy. Materials and methods: A total of 33 participants with DPN were recruited for the study based on inclusion criteria. Maximum plantar pressure was recorded using Win-Track 11K005, and the pain score was obtained using a visual analogue scale. Ten sessions of MaRhyThe® were given to all the participants. Outcome measures were evaluated at the baseline and after 10th session. Paired t-test was performed to analyze the changes in outcome measures. Results: Participants of DPN were recruited with the average age of 64 ± 9 years, and an average duration of diabetes was 14 ± 9 years were included. Results of the present study found significant improvement in neuropathic pain and plantar pressure in post intervention assessment. (p < 0.05). Conclusion: In the present study, we found that MaRhyThe® is effective in reducing neuropathic pain and maximum plantar pressure in type 2 diabetes mellitus with peripheral neuropathy.

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.).

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H2O2 in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.