Chiral Perovskite Nanocrystals for Asymmetric Reactions: A Highly Enantioselective Strategy for Photocatalytic Synthesis of N-C Axially Chiral Heterocycles.

Catalytic approaches to generate enantiospecific chiral centers are the major premise of modern organic chemistry. Heterogeneous catalysis is responsible for the vast majority of chemical transformations, yet the direct employment of chiral solid catalysts for asymmetric synthesis is mostly overlooked. Here, we demonstrated that a heterogeneous metal-halide perovskite nanocrystal (NC) catalyst is active for asymmetric organic synthesis under visible-light activation. Chiral 1-phenylethylamine (PEA)-hybridized perovskite PEA/CsPbBr3 NC photocatalysts exhibit an enantioselective (up to 99% enantiomer excess, ee) avenue to produce N-C axially chiral N-heterocycles, i.e., N-arylindoles from N-arylamine photo-oxidation. Mechanistic investigation indicated a discriminated prochiral binding of the N-arylamine substrates onto the chiral-NC surface with ca. -2.4 kcal/mol enantiodifferentiation. Our perovskite NC heterogeneous catalytic system not only demonstrates a promising strategy to address the long-term challenges in atroposelective pharmaceutical scaffold synthesis but also paves the road to directly employ chiral solids for asymmetric synthesis.

Synthesis and Characterization of Bimetallic Nanoclusters Stabilized by Chiral and Achiral Polyvinylpyrrolidinones. Catalytic C(sp3)-H Oxidation.

Second-generation chiral-substituted poly-N-vinylpyrrolidinones (CSPVPs) (-)-1R and (+)-1S were synthesized by free-radical polymerization of (3aR,6aR)- and (3aS,6aS)-5-ethenyl-tetrahydro-2,2-dimethyl-4H-1,3-dioxolo[4,5-c]pyrrol-4-one, respectively, using thermal and photochemical reactions. They were produced from respective d-isoascorbic acid and d-ribose. In addition, chiral polymer (-)-2 was also synthesized from the polymerization of (S)-3-(methoxymethoxy)-1-vinylpyrrolidin-2-one. Molecular weights of these chiral polymers were measured using HRMS, and the polymer chain tacticity was studied using 13C NMR spectroscopy. Chiral polymers (-)-1R, (+)-1S, and (-)-2 along with poly-N-vinylpyrrolidinone (PVP, MW 40K) were separately used in the stabilization of Cu/Au or Pd/Au nanoclusters. CD spectra of the bimetallic nanoclusters stabilized by (-)-1R and (+)-1S showed close to mirror-imaged CD absorption bands at wavelengths 200-300 nm, revealing that bimetallic nanoclusters' chiroptical responses are derived from chiral polymer-encapsulated nanomaterials. Chemo-, regio-, and stereo-selectivity was found in the catalytic C-H group oxidation reactions of complex bioactive natural products, such as ambroxide, menthofuran, boldine, estrone, dehydroabietylamine, 9-allogibberic acid, and sclareolide, and substituted adamantane molecules, when catalyst Cu/Au (3:1) or Pd/Au (3:1) stabilized by CSPVPs or PVP and oxidant H2O2 or t-BuOOH were applied. Oxidation of (+)-boldine N-oxide 23 using NMO as an oxidant yielded 4,5-dehydroboldine 27, and oxidation of (-)-9-allogibberic acid yielded C6,15 lactone 47 and C6-ketone 48.

Molecular genotyping of Indian blood group antigens amongst regular voluntary blood donors of Surat city, Gujarat, India.

BACKGROUND: There is paucity of data related to the prevalence of the rare blood group antigens amongst South Gujarat blood donor population due to unavailability and high cost of antisera. Therefore it is difficult to screen donors for such rare antigens by gold standard haemagglutination assay. The single nucleotide polymorphism (SNPs) of Ina and Inb antigens is the base of the PCR based detection methods that help to detect these alleles in regular voluntary blood donors. MATERIALS & METHODS: Blood samples of 200 unrelated regular voluntary blood donors wee collected. DNA was extracted using phenol-chloroform method and genotyped for Indian (Ina/IN*01, Inb/IN*02) blood group alleles by Sequence Specific PCR. Ina antigen positivity was confirmed by serology test. RESULTS: Four donors were found heterozygous for Ina antigen i.e. In (a + b+) by SS-PCR and their Ina positivity were confirmed by in-house polyclonal Anti-Ina reagent. SS-PCR was standardized using known heterozygous sample of a blood donor. The frequency of Ina antigen (2.0 %) was higher than Caucasians, lower than Iranians and Arabs while comparable to those reported among Indians of Mumbai city. CONCLUSION: In absence or unavailability of antisera particularly for low frequency alleles like Ina, such PCR based method would be extremely helpful to prepare rare donor registry by screening blood donors' at large scale. Red cells of Ina positive donors can be used as in-house reagent red cells for screening and identification of corresponding antibody.

Assessment of Long-Term in vitro Multiplied Human Wharton's Jelly-Derived Mesenchymal Stem Cells prior to Their Use in Clinical Administration.

The quantity of mesenchymal stem/stromal cells (MSCs) required for a particular therapy demands their subsequent expansion through ex vivo culture. During in vitro multiplication, they undergo replicative senescence which may alter their genetic stability. Therefore, this study was aimed to analyze cellular, molecular, and chromosomal alterations in Wharton's jelly-derived MSCs (WJ-MSCs) during their in vitro sequential passages, where WJ-MSCs were sequentially passaged up to P14 and cells were evaluated at an interval of P2, P6, P10, and P14. They were examined for their morphology, tumorigenicity, surface markers, stemness markers, DNA damage, chromosomal aberration, and telomere length. We have processed five full-term delivered human umbilical cord samples to obtain WJ-MSCs. Morphological appearance observed at initial stages was small fine spindle-shaped WJ-MSCs which were transformed to flat, long, and broader cells in later passages. The cell proliferation rate was gradually decreased after the 10th passage. WJ-MSCs have expressed stemness markers OCT-4 and NANOG, while they showed high expression of positive surface markers CD90 and CD105 and lower expression of CD34 and CD45. They were non-tumorigenic with slow cellular aging during subsequent passages. There was no chromosomal abnormality up to the 14th passage, while increase in comet score and decrease in telomere length were observed in later passages. Hence, our study suggests that early and middle passaged (less than P10) WJ-MSCs are good candidates for clinical administration for treatment.

Risk of hepatitis-E virus infections among blood donors in a regional blood transfusion centre in western India.

BACKGROUND: Hepatitis-E virus (HEV) is an emerging infectious threat to blood safety. The enormity of the transmission of HEV and its clinical consequence are issues currently under debate. This study aimed to evaluate the prevalence of HEV-RNA in blood donors in western India. MATERIALS AND METHODS: We screened 13 050 blood donors for HEV using HEV-RNA screening of 10 mini-pools using RealStar HEV RT-PCR Kit (95% limit of detection (LOD): 4.7 IU/ml). Furthermore, all HEV-RNA-positive donors were investigated for the presence of IgM/IgG antibody along with liver function tests. RESULTS: Of the 13 050 blood donations, 7 (0.53%) were found to be HEV-RNA positive, and the prevalence of HEV nucleic acid testing yield cases among blood donors was 1 in 1864. All seven HEV-RNA-positive samples were tested with anti-HEV IgM and anti-HEV IgG antibodies; this resulted in two (28.5%) positive anti-HEV IgM and two (28.5%) positive anti-HEV IgG antibodies. Hepatic activity was measured, with two of seven HEV-RNA-positive donors demonstrating abnormal serum glutamic oxaloacetic transaminase (SGOT) andserum glutamic pyruvic transaminase (SGPT). Two HEV-RNA-positive blood donors who had abnormal SGOT and SGPT were found to have a high HEV viral load. Furthermore, we were able to follow up two HEV-RNA donors, and both were HEV-RNA positive and had anti-HEV IgM and anti-HEV IgG antibodies; moreover, their liver function tests were also abnormal. One of the HEV-RNA donors with high viral load did show hepatitis-E-like virus on electron microscopy. CONCLUSION: Our studies indicate that there is a significant risk of blood-borne transmission of HEV. This finding may help to provide a direction towards the safety of blood transfusions in clinical settings in countries like India, which fall under the endemic category for HEV infection.

Comparison of serology and molecular detection of common red cell antigens in multitransfused thalassemia major and sickle cell disease patients.

BACKGROUND: Serological testing for extended RHCcEe, Kell, Kidd and Duffy blood grouping from multitransfused patients may not give correct blood grouping of the recipient. Hence molecular testing for these blood groups was compared with serological groups in a cohort of multitransfused thalassemia mjor and sickle cell anaemia patients. OBJECTIVE: Molecular genotyping of antigens of Rh (D, C, c, E, e), Kell (K, k), Duffy (Fya, Fyb) and Kidd (Jka, Jkb) blood group antigens by PCR and PCR-RFLP methods and comparison of predicted genotypes with their serological phenotypes. MATERIALS AND METHODS: A cohort of multitransfused thalassemia and sickle cell anemia patient were serologically and molecularly tested for RHCc, RHEe, K, k Fya, Fyb, Jka and Jkb antigens and compared. Serological testing was done by tube agglutination and molecular testing was done either by allele specific PCR or by RFLP technique just before next transfusion. RESULTS: In more than 80% of the cases recipient's molecular testing blood groups were at variance with serologically tested blood groups (p < 0.0001). Mixed field reactions in serological typing were common. In sickle cell anemia patients no discrepancy was found. Molecular technique results were checked by Sanger's sequencing. DISCUSSION: Extended phenotyping in multitransfused thalassemia patients by serological technique often donot detect the exact red cell phenotype of the recipient and molecular techniques for such grouping is preferable, especially in multitransfused thalassemia patients where red cells from previous transfusions continues to be present in significant numbers whenever the testing is done.

Support-Free Pd3 Co NCs as an Efficient Heterogeneous Nanocatalyst for New Organic Transformations of C-C Coupling Reactions.

A support-free heterogeneous Pd3 Co nanostructured composite (NC), synthesized through a hydrothermal route, acted as an effective catalytic system in multivariate Heck-, Sonogashira-, and Suzuki-type coupling reactions of iodonium ylides. The XPS analysis of the bimetallic Pd3 Co NCs confirmed the elemental composition as 75 % palladium and 25 % cobalt. Furthermore, high-resolution (HR) TEM analysis confirmed the spherical morphology of the Pd3 Co bimetallic nanoparticles. The average diameter of the NCs is 14.8 nm. The coupling reaction proceeded through the generation of α-iodoenones with simultaneous migration of the phenyl group, thereby giving a scaffold with higher atom economy. The heterogeneous Pd3 Co NCs were recycled and reused without any significant change in catalytic ability for up to five reaction cycles. The high concentration of Pd and association of cobalt into the lattice of palladium appears to enhance its catalytic ability for the diverse coupling reactions in comparison with its monometallic counterparts as well as with bimetallic NCs with a comparatively lesser amount of Pd.

Anti tissue transglutaminase antibody in idiopathic autoimmune haemolytic anemia.

BACKGROUND: In idiopathic autoimmune haemolytic anaemia (AIHA haemolytic antibodies are directed to every type of red cellsWestern blot studies have shown antibody positivity towards red cell anion channel complex which also includes band 4.2 a protein with similarities to tissue trans glutaminase. OBJECTIVE: Evaluation of AIHA for anti tissue transglutaminase antibody (Anti tTG). MATERIALS & METHODS: Twenty three AIHA patients were tested along with routine hamatogical work up, for a series of auto antibodies and red cell eluates and serum from the patents were tested against solubilised group O red cell ghosts on western blot. Other ancillary investigations were done to rule out complications and secondary causes of haemolysis. RESULTS: 11/23 patients (48%) were positive for anti tTG, Four, 3 and 8,7 patients were positive for anti thyroid, anti b2 glycoprotein, lupus anticoagulant and ANA respectively. One patient with anti tTG had biopsy proven celiac disease. Three patient developed DVT and all of them were lupus anticoagulant as well as b2 gp-1 antibody positive.17 had become Coombs test negative on treatment while 21/23 had positive western blot test. DISCUSSION & CONCLUSION: There is strong association of anti tTG antibody with idiopathic AIHA. Aetiological association of this finding needs exploration.

Comparative study of alloimmunization against red cell antigens in sickle cell disease & thalassaemia major patients on regular red cell transfusion.

BACKGROUND & OBJECTIVES: : Sickle cell disease (SCD) patients require red cell transfusion during different clinical complications of the disease. Such patients are at a high risk for developing alloantibody against red cell antigens. From India, there are limited data available on alloantibody formation in multiply transfused SCD patients. The present study was thus undertaken to fill up this lacunae by looking at the development of red cell alloantibodies in SCD and ß-thalassaemia patients on regular transfusion. METHODS: : All sickle cell disease patients undergoing red cell transfusion between 2008 and 2016, were included. During this period, a large number of ß-thalassaemia major patients also underwent regular red cell transfusion. These thalassaemia patients were also included to compare the tendency of antibody formation between SCD and ß-thalassaemia major patients. All patients before regular transfusion were regularly assessed for the development of red cell antibody. Red cell antigen, antibody screen crossmatch and antibody identification were done using the standard technique. RESULTS: : A total of 138 patients with SCD aged between 4 and 53 yr (mean 17.6 yr) consisting of 83 males and 55 females (male:female, 1.5:1) along with 333 transfusion-dependent ß-thalassaemia patients were studied. Over the last eight years, 15 patients with SCD and four patients with thalassaemia developed alloantibody (P <0.001). Antibody specificity of their alloantibodies was against Rhc, RhE, Kell, Fya and Fyb only. Sickle cell disease patients with and without alloantibody required on the average 11.8 and 8.6 units of red cell concentrate, respectively (P <0.05). INTERPRETATION & CONCLUSIONS: : About 11 per cent of the transfused sickle cells patients developed alloantibodies. The antibody specificity was restricted to Rh, Kell and Duffy blood group systems. Extended antigen matching involving Rh, Kell and Duffy antigens may prevent alloantibody in such patients.

Sensitive and selective fluorometric determination of DNA by using layered hexagonal nanosheets of a covalent organic framework prepared from p-phenylenediamine and benzene-1,3,5-tricarboxaldehyde.

A modified method is described for the preparation of amino-functionalized covalent organic framework nanosheets (COF-NSs). These consist of hexagonal layered sheets and were prepared from commercially available starting materials (p-phenylenediamine and benzene-1,3,5-tricarboxaldehyde). The interlayer stacking interactions between the ultra-thin COF-NSs became weak because the π stacking is destroyed by sonication. This result in the exfoliation of COF-NSs. As an application, the COF-NSs used for sensitive and selective fluorometric determination of DNA. To reach this goal, H1 and H2 hairpin-like DNA probes were chosen; H1 used Texas Red-labeled dye as a fluorescent probe. The addition of the COF-NSs, the hairpin probes was adsorbed onto the porous surface of the COFNSs. The π stacking and hydrogen-bond interactions between COFNSs and nucleic acid quench the fluorescence of the Texas red-labeled probe. The target DNA enables the recovery of the quenched fluorescence of the Texas red-labelled probe by triggering an inter-chain hybridization within hairpin probes. This results in a weaker interaction of double-stranded DNA (dsDNA) with the COFNSs. Consequently, the dsDNA detaches from the COFNSs, thereby recovering the dye's fluorescence (excitation/emission maxima at 590/612 nm) with increasing target DNA concentration. The findings were applied to design a method for the determination of DNA that has a 2 pM detection limit. This is significantly lower than the limit of detection reported previously for 2D nanomaterial-based fluorometric DNA assays. Graphical abstractSchematic representation of 2D-covalent organic framework nanosheets (COF-NSs) probe act as a quencher allowing the highly sensitive and selective fluorescence turn-on detection for biomolecules. Here the H1 H2 are hairpin DNAs. H1 is associated with the fluorescent tag (red circle), while the "fluorescence off" state it denoted as a black circle.

Pattern of distribution of 35 red cell antigens in regular voluntary blood donors of South Gujarat, India.

BACKGROUND: Extended phenotyping is one of the important method of reducing red cell alloimmunisation. Extended phenotyping of red cells from voluntary donors have many uses in addition to its application in population genetics. As there was very little data extended phenotyping on a cohort of Indian Voluntary blood donors this project was undertaken. STUDY DESIGN & METHODOLOGY: 200 regular voluntary blood donors having 'O' blood group were included for red cell antigen typing of Rh (D,C,E,c,e), Kell (K, k, Kpa, Kpb), Duffy (Fya, Fyb), Kidd (Jka, Jkb), Lewis(Lea, Leb), P(P1), MNS (M, N,S,s), and Lutheran (Lua, Lub), Colton (Coa, Cob), Diago (Diaa, Wra), Vw and Xga antigens using conventional antisera provided by DIAGAST. Calculations of antigen and phenotypes frequencies were expressed as percentages. RESULTS: Out of 200 'O' group blood donors, 96.5% were Rh D and 2.5% were K positive. Amongst Rh antigens, e was the most common (100%) followed by D, C (91.0%), c (50.5%) and E (16.5%) with DCe/DCe (R1R1, 48.0%) being the most common phenotype. In Kell blood group system, we found k antigen to be 100% and a rare phenotype Kp (a + b+) was found in 1% of the donors. For Kidd and Duffy blood group systems, Jk (a + b+) and Fy (a + b-) were the most common phenotypes (39.0% and 64.0%, respectively). In the MNS blood group system, M + N+ (67.5%) and S + s+(43.5%) were the most common phenotypes. There were antigens like Cw(3.5%), K(2.3%), Kpa(1.2%), Ina(1.0%), Vw(1.2%), Coa(4.5%), Cob(1%), Lua(1.75%), Dia+(1.2%), and Wra+(0.6%) with frequency < 5% in the donor population. CONCLUSION: Extensively antigen phenotypes group 'O' red cells showed significant variation with other population from India as well as with Caucasian and black population. Extensive phenotyping 'O' group regular blood donors of red cell antigens is very useful to prepare in-house red cell panels for identification of alloantibodies.

A multiplex ARMS PCR approach to detection of common ß-globin gene mutations.

BACKGROUND: ß-thalassaemia is a group of inherited single-gene disorders worldwide. Each ethnic population has its own common mutations. Heterogeneity of ß-thalassaemia mutations in multi-ethnic population of Surat, makes molecular diagnosis expensive and time consuming. METHODS: Specific primers were used to differentiate four common mutations, IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G), by a simple PCR involving a multiplex amplification refractory mutation system. RESULTS: Several high prevalence ß-Thalassemia trait groups constituted by Muslims, Patels, Sindhis, ModhBanias, and Mahayavanshi. Four most common mutations detected in them are IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G). We identified each of these ß-thalassemia mutations in multiplexed ARMS from positive control samples. Our multiplex-ARMS-PCR system was first standardized on positive DNA samples with above known four most common ß-thalassemia mutations, and these positive samples had been diagnosed with ß-thalassemia and also all these samples belonged to Surat ethnic groups. The system was subsequently tested on 110 blood samples from different ethnic backgrounds with unknown ß-thalassemia mutations which were in all specimens. CONCLUSION: The ARMS multiplex system was found reliable, cost effective, fast and most applicable for mutation screening of Thalassemia in Surat populations.

NAT positivity in seronegative voluntary blood donors from western India.

BACKGROUND AND OBJECTIVES: Prevalence and composition of Hepatitis B, Hepatitis C and HIV-1, NAT positive but seronegative voluntary blood donors from western part of India is yet to be documented. MATERIAL AND METHODS: Over last 2 1/2 years all the seronegative voluntary blood donors were tested using 10 minipools on a semiautomated NAT testing platform. The positively tested donors were followed up for at least five months for development of seropositivity. RESULTS: 79532 seronegative donations were tested by 10 minipool (MP) NAT leading to 51 positive sample (44 Hep B, 5 HIV 1 and Hep C positive). All the HIV and Hep C NAT positive donors eventually developed seropositivity and out of 44 Hep B NAT positive donors, 31 developed seropositivity within six months of follow up, following counseling of the donors. This data translate into NAT yield of 1:1559 donors for all virus taken together. NAT yield for Hep B 1:1807 donors were much higher than HIV 1 in 1:15906 and HCV yield of 1:39761. Semiautomated minipool NAT testing system was found to be cost effective way for improving blood safety. INTERPRETATION AND CONCLUSION: Seronegative NAT yield in voluntary blood donors are quiet high in western part of India and in line with rest of the country is mainly due to Hepatitis B infection. Implementation of strict donor screening, Hep B vaccination of the population and sample mutation of NAT testing should be under taken on war footing.

Assessment of semi-automated nucleic acid testing programme in a Regional Blood Transfusion Centre.

BACKGROUND: Detection of human immunodeficiency virus type-1 (HIV-1), hepatitis-C (HCV) and hepatitis-B virus (HBV) in the blood donors is crucial. An efficient form of detection is nucleic acid testing (NAT) in blood screening. We assessed the suitability of commercial NAT testing in a developing country, focusing on the Altona RealStar assay and the method of Sacace Biotechnologies. METHODS: We have standardised and validated commercially available NAT kits with a semi-automated system for detection of HBV, HCV and HIV-1 in blood donations. The MP-NAT (mini-pool) assay consists of pooling of sample, virus extraction, amplification and detection with commercially available NAT kits. An internal control (IC) is incorporated in the assay to monitor the extraction, target amplification and detection process. RESULTS: The sensitivity of the Altona RealStar assay at 10-MP for each viral target was evaluated, HBV showed amplification in all diluted positive samples of 100, 50, 25, 10 and 5 IU/ml. HIV and HCV infected samples showed amplification in all diluted positive samples of 500, 100, 50 and 30 IU/ml. For HIV, out of six diluted samples of 30 IU/ml, five were amplified. A total of 14,170 seronegative blood samples were tested by RealStar PCR kit in 10-MP and 6 (0.042%) samples/pools were positive. A total of 65,362 seronegative blood donations were also tested by kits of Sacace Biotechnologies, in 10-MP and 45 (0.075%) pools were positive. The prevalence of combined NAT yield cases among routine donors was 1 in 1559 donations tested for all the 3 viruses. CONCLUSION: The semi-automated combined system for NAT screening assays is robust, sensitive, reproducible, and this gives an additional layer of safety with affordable cost.

SOCS3 induces neurite differentiation and promotes neuronal cell survival.

Cytokines and growth factors play an important role in neuronal survival as well as cell death. The family of suppressors of cytokine signalling (SOCS) proteins, which includes SOCS1-7 and cytokine-induced suppressor (CIS), has been shown to act as negative regulators of cytokine-induced signalling. In this report, we highlight the role of SOCS3 in regulating neuronal differentiation and survival. We observed increased SOCS3 expression upon differentiation of PC12 cells as well as neural stem cells. SOCS3 overexpression upregulated differentiation of both neural stem cells and PC12 cells even in the absence of NGF, as evidenced by enhanced neurite outgrowth and upregulation of GAP43, marker associated with neurite outgrowth. siRNA-mediated silencing of SOCS3 confirmed the potential role of SOCS3 in neuritogenesis. We observed that, SOCS3-induced neurite differentiation was mediated via the PI3 kinase pathway. Another interesting observation was that SOCS3 overexpression promoted neuronal cell survival under H2 O2 -mediated stress indicating its fundamental role in cell survival. In conclusion, our results indicate that SOCS3 promotes differentiation and survival of neural cells and could be potentially useful in future therapy for treatment of neurodegenerative disorders. © 2016 IUBMB Life, 68(6):468-476, 2016.

Confirmatory Assay for Laboratory Diagnosis of Malaria Using Molecular Approach.

BACKGROUND: Prompt malarial treatment and surveillance is crucial for accurate diagnosis of Plasmodium Sp. Gold standard microscopic examination has been widely applied for diagnosis of malaria in most part of the endemic areas. But in case of submicroscopic and asymptomatic microscopic diagnosis is questioned. The study aims to develop a simple, cost effective & robust nucleic acid amplification technique for the detection of malaria parasite. METHODS: Study population included 50 clinically diagnosed positive malaria patient samples from various pathological laboratories. Microscopy by preparing thick film was carried out of every sample for primary screening in the available facility of Surat Raktadan Kendra & Research Centre- Blood Bank. The conventional PCR (Polymerase Chain Reaction) was applied for genus-specific amplification targeting the 18 S rRNA gene of Plasmodium. Agarose gel electrophoresis was used to separate and analyze the amplified PCR product using 2% Agarose gel. RESULTS AND CONCLUSION: The study shows that nested PCR not only detected all microscopic positive samples, but also detected submicroscopic infections that were missed or misread by microscopy. Hence, the sensitivity of molecular based detection technique is proved to be more compared to microscopic examination.

Nyctanthes arbor-tristis alkaloids activates p53 independent cell death receptor and necroptosis pathways in HepG2 cells.

Nyctanthes arbor-tristis is a traditional medicinal plant with potential anti-cancer properties. In this study, crude and alkaloid extracts were prepared from different parts of the plant, and their cytotoxicity was evaluated on four different cancer cell lines. The alkaloid extracts from the leaf and fruit showed promising results, with the HepG2 cell line exhibiting significant cytotoxicity. The promising extracts were further studied for their apoptotic potential using various methods, including DNA fragmentation, TUNEL, Caspase-3 activity, Giemsa, and Hoechst staining. Our results indicated that the fruit extract had the highest apoptotic potential, with clear nuclear condensation, fragmentation, and apoptotic bodies observed. We also investigated the alteration of the Bax/Bcl-2 ratio both at the mRNA and protein levels. Our results showed a significant upregulation of the Bax gene and downregulation of the Bcl-2 gene for the fruit alkaloid extract. This indicates that the phenomenon of cell death expression might be following a p53-independent extrinsic pathway and Bax-activated caspase-independent AIF-mediated necroptosis in the HepG2 cancer cell line. Overall, our findings suggest that Nyctanthes arbor-tristis has potential as a therapeutic option for cancer treatment. The alkaloid extracts from the leaf and fruit may hold promise as a source of bioactive compounds for further development into anti-cancer agents. Further studies are needed to explore the underlying mechanisms of their cytotoxic and apoptotic effects and to evaluate their safety and efficacy in animal models and clinical trials.

Seroprevalence of HBV, HCV and HIV-1 and Correlation with Molecular Markers among Multi-Transfused Thalassemia Patients in Western India.

BACKGROUND: Multitransfused ß-thalassemia major patients are always at high risk of having Transfusion Transmitted Infections (TTIs). This study was aimed to determine the seroprevalence of HBsAg, Anti-HIV-1/2, and Anti-HCV among these patients and to correlate the same with NAT testing. METHODS: A total of 196 patients with ß-thalassemia were included in the study. Patients were screened for the presence of viral markers by third-generation ELISA test as well as for viral DNA/RNA by NAT test. RESULTS: Among 196 multi-transfused Beta-thalassemia patients, the seroprevalence of anti-HCV was very high 100 (51.1%), however, anti-HIV1/2 was 6 (3.1%), and HBsAg were 3 (1.5%). Surprisingly similar patterns were observed in the prevalence of molecular markers, as HCV-RNA were 66 (33.7%) of the patients along with HIV-1 RNA were 8 (4.1%), and HBV-DNA were 5 (2.5%) patients. Overall eight (4.1%) patients were found to have coinfections, where two were positive for HBsAg/anti-HCV by ELISA along with 3 (1.5%) were positive for HBV-DNA/ HCV-RNA, 1 (0.5%) was positive for HIV-RNA/HBV-DNA, and 2 (1%) had coinfection of HIV-RNA/ HCV RNA by NAT testing. CONCLUSION: The prevalence of HCV infection among multi-transfused ß-thalassemia patients is significantly higher than that of the HBV and HIV infections. This scenario should be controlled and monitored by doing regular follow-up testing schedules of such patients and also the administration of the booster dose of the HBV vaccine along with HCV treatment with antiviral DAAs.