Redefining venereology practice in Tamil Nadu, South India - Nakshatra Health - A networking model.

Background: Accessing care for sexual health has always been a challenge in our Indian context. The primary reason is a lack of awareness of modes of transmission of sexually transmitted diseases (STD), appropriate testing, and treatment options. The second is taboo associated with the morality of the individual. The third is the accessibility and availability of Quality STD care by allopathic specialists in comparison to the demand. This has paved way for lots of myths and misconceptions among the general public regarding STDs and HIV disease. Compounding it is the mushrooming of nonqualified practitioners and healers who claim to cure all STDs and HIV has led to poor treatment outcomes. Several methods of partnership with qualified allopathic practitioners have been tried for the provision of quality STD care by various donor-funded, HIV and STD prevention programs in the country. The key reasons for the nonsustainability of these clinics were the lack of technical capability to handle the sexual health and STD clinical cases and the huge cost involved in the advertisement and maintenance of the clinics. Methodology: Seven clinicians from different geographical locations in Tamil Nadu, who were exclusively qualified in Venereology, conceived the idea of provision of comprehensive clinical care cum counseling and testing services through a networking model from December 01, 2020. The model comprised the following: (1) Dedicated YouTube channel (Nakshatra Health) to provide scientific information on STD and HIV, modes of transmission, clinical symptoms, lab testing, interpretation of results, clinical treatment options, and counseling on prevention, (2) Dedicated telephone helpline was established with WhatsApp to answer the queries of clients by the network venereologists, (3) Dedicated website (www.nakshatra.health) was developed to provide information on STD and HIV and options to clarify doubts and fix appointments online, (4) Clinical consultations were done in a hybrid mode with an option for direct clinic and online consultation. Prescriptions were provided using dedicated software instantly, (5) Networking was done with NABL-accredited labs and collection centers for performing STD and HIV tests with e-test request forms, (6) Networking was done with pharma companies to provide pre- and postexposure prophylaxis (PEP) services to clients through e-prescriptions. Cross referrals were made across the network members to facilitate easy access to clinical services by clients from different parts of Tamil Nadu. The entire concept was branded as "Nakshatra health" with a tagline - "Your sparkling solution for safe sexual health care." Quality STD care and ethical practice were the underlying motos of this concept. Results: During the 20 months (December 2020 to July 2022), 6442 phone calls and 9328 WhatsApp messages were received. 82.3% of the calls and messages were queries from clients related to their sexual exposures, and 16.4% were general information seekers. During this period, the Nakshatra Health YouTube channel had 1590 subscribers and nearly 2.4 lakh views of all its 24 videos. Among the viewers, 92.4% were male. 52.29% of viewers were between the ages of 25 and 34 years, 28.25% were between the ages of 18 and 24 years, and 17.25% viewers were between the ages of 35 and 44 years. 86% of the viewers were from India and 13% were Tamil-speaking viewers from Middle East, Southeast Asian countries and 1% were from European, African, and American countries. The most commonly watched videos were related to HIV symptoms and lab tests for STD and HIV. 16% of the viewers repeatedly watched the various videos in the series. The network laboratories provided testing services for 1082 clients with 2423 various STD/HIV tests. Totally 3328 clients availed of online consultation and 924 clients accessed clinic-based services across the network members. Among these cases, 18 cases of Syphilis (primary and secondary) and 12 cases of acute gonococcal urethritis, and 10 cases of genital warts were diagnosed and treated. 12 cases of phimosis and 4 cases of anal warts were surgically treated in collaboration with a surgeon. Through this initiative, PEP and PreP services were provided to 228 and 8 individuals. Conclusion: Designing a comprehensive sexual health service package is crucial to ensure the availability and ease of access to services to the general public. Provision of correct scientific information, networking and cross-referral of cases with like-minded dermatovenereologists/clinicians interested in venereology sexually transmitted infections, easily accessible clinical, laboratory, and treatment services including PreP and PEP medications, and ethical practice are the key factors for scaling this concept.

A Unique Metro Choice Behaviour of Suburban Rail Passengers in India.

This study aims to analyse the inter-rail modal shift behaviour of suburban rail passengers to examine ridership for the proposed metro extension corridor in Chennai, India. This investigation was conducted in 2019 as part of a feasibility study for the extension of the metro line spanning between Chennai Airport and Kilambakkam, a southern suburb of Chennai. The same origin-destination pair is also served by the suburban rail system. It is an extension of the operating line from Washermenpet to the airport of the Phase I metro project. For this inter-rail competition study, a sample of 272 suburban rail passengers covering work, education and other trip purposes were interviewed using a stated preference questionnaire. Six stated scenarios were considered for analyses which included travel time saving by using the metro along with the fare difference between metro and bus. The study revealed that suburban rail passengers were less concerned about travel time saving and gave priority to fare difference irrespective of trip purpose. This shows the unique metro choice behaviour of suburban rail travellers in the Indian context.

Clinicomycological study of vulvovaginal candidiasis.

BACKGROUND: Vaginal candidiasis is one of the most common vaginal infections in women of reproductive age group caused by Candida species of fungi, a normal inhabitant of vagina. AIMS: The aim is to study the culture characteristics, species prevalence, and drug sensitivity of vulvovaginal candidiasis (VVC) among females with genital pruritus and discharge per vaginum who are proved to have Candida infection by a vaginal smear study. MATERIALS AND METHODS: A prospective study was done on female patients aged 18 years attending the Sexually Transmitted Diseases Clinic of the Department of Dermato-Venereo-Leprology (DVL), Tirunelveli Medical College, with complaints of genital pruritus and vaginal discharge over a period of 1½ years from 2013 to 2015. After clinical examination, vaginal discharge was smeared for wet mount, potassium hydroxide mount, and Gram stain. Those with pseudohyphae were chosen for culture, speciation, and drug sensitivity. RESULTS: The most common age group affected was 18-35 years. The most common predisposing factor was HIV infection-induced immunosuppression (48.7%), followed by antibiotic usage and diabetes mellitus. Clinical presentation of VVC was similar in HIV-positive and HIV-negative patients. Recurrent VVC makes up 12% of total cases with most common species being Candida albicans whereas Candida glabrata among the non-C. albicans. Drug sensitivity pattern of C. albicans showed the highest sensitivity to nystatin 85.7%, followed by triazoles (75%-85%) with ketoconazole being more resistant (40%-60%), and was similar in C. albicans and non-C. albicans. LIMITATIONS: Exclusion of unmarried and pregnant women and lack of follow-up were limitations in our study. CONCLUSIONS: According to our study, elimination of predisposing factors, speciation of fungus, and treatment based on drug sensitivity pattern will reduce the incidence of VVC.

Coumarin polycaprolactone polymeric nanoparticles: light and tumor microenvironment activated cocktail drug delivery.

Highly sensitive hypoxia (H2O2)-activated photoresponsive polymeric nanoparticles for cocktail delivery of anticancer drugs doxorubicin (Dox) and chlorambucil (Cbl) were developed. The photoresponsive polymer conjugate was constructed by ring-opening polymerization (ROP) of caprolactone (as the tail) with 7-hydroxycoumarin chlorambucil (as the head). During nanoprecipitation, the polycaprolactone chain wrapped around the hydrophobic core (coumarin chlorambucil) to form a "shell". Interestingly, the polycaprolactone-tagged coumarin-chlorambucil (PCL-CC) NPs provided sufficient space for co-encapsulation of another hydrophobic anticancer drug Dox with a loading efficiency of 13 wt%. The controlled release of Dox and Cbl from Dox-PCL-CC NPs was investigated under three different conditions: (i) in the presence of H2O2 (tumor microenvironment), (ii) photoirradiation using UV light of ≥365 nm for 60 min, and (iii) photoirradiation using UV light of ≥365 nm for 15 min in the presence of H2O2. Results showed that photoirradiation in the presence of H2O2 results in generation of reactive oxygen species (HOO-, OH-), which assist hydrolysis of the ester group in the polymeric backbone of Dox-PCL-CC NPs and UV irradiation leads to cleavage of the coumarin-chlorambucil ester linkage, leading to burst release of Dox and Cbl. The drug release profile from the NPs under three different conditions was monitored by different instrumental techniques, e.g. emission spectroscopy, MALDI-Tof mass spectrometry, DLS and HPLC analysis. In vitro biological studies revealed that the present system can efficiently deliver the cocktail anticancer drugs with full control over release into the tumor cells by means of H2O2 and light activation.

Excavating the Role of Aloe Vera Wrapped Mesoporous Hydroxyapatite Frame Ornamentation in Newly Architectured Polyurethane Scaffolds for Osteogenesis and Guided Bone Regeneration with Microbial Protection.

Guided bone regeneration (GBR) scaffolds are unsuccessful in many clinical applications due to a high incidence of postoperative infection. The objective of this work is to fabricate GBR with an anti-infective electrospun scaffold by ornamenting segmented polyurethane (SPU) with two-dimensional Aloe vera wrapped mesoporous hydroxyapatite (Al-mHA) nanorods. The antimicrobial characteristic of the scaffold has been retrieved from the prepared Al-mHA frame with high aspect ratio (∼14.2) via biosynthesis route using Aloe vera (Aloe barbadensis miller) extract. The Al-mHA frame was introduced into an unprecedented SPU matrix (solution polymerized) based on combinatorial soft segments of poly(ε-caprolactone) (PCL), poly(ethylene carbonate) (PEC), and poly(dimethylsiloxane) (PDMS), by an in situ technique followed by electrospinning to fabricate scaffolds. For comparison, pristine mHA nanorods are also ornamented into it. An enzymatic ring-opening polymerization technique was adapted to synthesize soft segment of (PCL-PEC-b-PDMS). Structure elucidation of the synthesized polymers is established by nuclear magnetic resonance spectroscopy. Sparingly, Al-mHA ornamented scaffolds exhibit tremendous improvement (175%) in the mechanical properties with promising antimicrobial activity against various human pathogens. After confirmation of high osteoconductivity, improved biodegradation, and excellent biocompatibility against osteoblast-like MG63 cells (in vitro), the scaffolds were implanted in rabbits as an animal model by subcutaneous and intraosseous (tibial) sites. Improved in vivo biocompatibilities, biodegradation, osteoconductivity, and the ability to provide an adequate biomimetic environment for biomineralization for GBR of the scaffolds (SPU and ornamented SPUs) have been found from the various histological sections. Early cartilage formation, endochondral ossification, and rapid bone healing at 4 weeks were found in the defects filled with Al-mHA ornamented scaffold compared to pristine SPU scaffold. Organ toxicity studies further confirm the absence of appreciable tissue architecture abnormalities in the renal hepatic and cardiac tissue sections. The entire results of this study manifest the feasibility of fabricating a mechanically adequate tailored nanofibrous SPU scaffold based on combinatorial soft segments of PCL, PEC, and PDMS by a biomimetic approach and the advantages of an Aloe vera wrapped mHA frame in promoting osteoblast phenotype progression with microbial protection for potential GBR applications.

On-Demand Guided Bone Regeneration with Microbial Protection of Ornamented SPU Scaffold with Bismuth-Doped Single Crystalline Hydroxyapatite: Augmentation and Cartilage Formation.

Guided bone regeneration (GBR) scaffolds are futile in many clinical applications due to infection problems. In this work, we fabricated GBR with an anti-infective scaffold by ornamenting 2D single crystalline bismuth-doped nanohydroxyapatite (Bi-nHA) rods onto segmented polyurethane (SPU). Bi-nHA with high aspect ratio was prepared without any templates. Subsequently, it was introduced into an unprecedented synthesized SPU matrix based on dual soft segments (PCL-b-PDMS) of poly(ε-caprolactone) (PCL) and poly(dimethylsiloxane) (PDMS), by an in situ technique followed by electrospinning to fabricate scaffolds. For comparison, undoped pristine nHA rods were also ornamented into it. The enzymatic ring-opening polymerization technique was adapted to synthesize soft segments of PCL-b-PDMS copolymers of SPU. Structure elucidation of the synthesized polymers is done by nuclear magnetic resonance spectroscopy. Sparingly, Bi-nHA ornamented scaffolds exhibit tremendous improvement (155%) in the mechanical properties with excellent antimicrobial activity against various human pathogens. After confirmation of high osteoconductivity, improved biodegradation, and excellent biocompatibility against osteoblast cells (in vitro), the scaffolds were implanted in rabbits by subcutaneous and intraosseous (tibial) sites. Various histological sections reveal the signatures of early cartilage formation, endochondral ossification, and rapid bone healing at 4 weeks of the critical defects filled with ornamented scaffold compared to SPU scaffold. This implies osteogenic potential and ability to provide an adequate biomimetic microenvironment for mineralization for GBR of the scaffolds. Organ toxicity studies further confirm that no tissue architecture abnormalities were observed in hepatic, cardiac, and renal tissue sections. This finding manifests the feasibility of fabricating a mechanically adequate nanofibrous SPU scaffold by a biomimetic strategy and the advantages of Bi-nHA ornamentation in promoting osteoblast phenotype progression with microbial protection (on-demand) for GBR applications.

Synthesis and Characterization of Novel Polycarbonate Based Polyurethane/Polymer Wrapped Hydroxyapatite Nanocomposites: Mechanical Properties, Osteoconductivity and Biocompatibility.

The present investigation reports the preparation of two types of 2D rod-like nano-hydroxyapatite (nHA) (unmodified and Polypropylene glycol (PPG) wrapped) of varying high-aspect ratios, by modified co-precipitation methods, without any templates. These nHA were successfully introduced into novel synthesized Thermoplastic Polyurethane (TPU) matrices based on polycarbonate soft segments, by both in-situ and ex-situ techniques. Physico-mechanical properties of the in-situ prepared TPU/nHA nanocomposites were found to be superior compared to the ex-situ counterparts, and pristine nHA reinforced TPU. Improved biocompatibility of the prepared nanocomposites was confirmed by MTT assays using osteoblast-like MG63 cells. Cell proliferation was evident over an extended period. Osteoconductivity of the nanocomposites was observed by successful formation of an apatite layer on the surface of the samples, after immersion into simulated body fluid (SBF). Prothrombin time (PT) and activated partial thromboplastin time (APTT), as calculated from coagulation assays, displayed an increase in the clotting time, particularly for the PPG-wrapped nHA nanocomposites, prepared through the in-situ technique. Only 0.3% of hemolysis was observed for the in-situ prepared nanocomposites, which establishes the antithrombotic property of the material. The key parameters for enhancing the technical properties and biocompatibility of the nanocomposites are: the interfacial adhesion parameter (B(σy)), the polymer-filler affinity, the aspect ratio of filler and non-covalent modifications, and the state of dispersion. Thus, the novel TPU/polymer wrapped nHA nanocomposites have great potential for biomedical applications, in particular for vascular prostheses, cardiovascular implants, scaffolds, and soft and hard tissues implants.

Temporal distribution and behaviour of sand flies (Diptera: Psychodidae) in a cutaneous leishmaniasis focus of the Kani Tribe settlements in the Western Ghats, India.

The temporal distribution of sand flies in relation to environmental factors was studied in the Kani tribe settlements located on the southernmost part of the Western Ghats, Kerala, India, between June 2012 and May 2013. This area is known for occurrence of cutaneous leishmaniasis (CL) cases. Employing hand-held aspirator, light trap and sticky-trap collection methods, a total of 7874 sand fly specimens, comprising 19 species was collected. Sergentomyia baghdadis was predominant species, followed by Phlebotomus argentipes. Sand fly abundance was significantly higher indoors (χ(2)=9241.8; p=0.0001) than outdoors. Mean density of P. argentipes in human dwellings, cattle sheds and outdoors was 7.2±2.9, 27.33±21.1 and 0.64±0.2 females/per man-hour (MHR), respectively. No sand fly species other than P. argentipes was obtained from cattle sheds. Although, sand fly populations were prevalent throughout the year, their abundance fluctuated with seasonal changes. Multiple regression analysis with backward elimination indicated that the increase in precipitation and relative humidity contributed to a significant positive association with the increase in sand fly abundance, while the increase in temperature showed no association. Fully engorged female sand flies tested for blood meal source showed multiple host-blood feeding. Analysis of resting populations of sand flies collected from human shelters indicated that the populations were found maximum on interior walls at 6-8 and >8 ft height, including ceiling during summer (F=83.7, df=6, p=0.001) and at the lower half of the wall at 0 and 0-2 ft height, during monsoon season (F=41.4, df=6, p=0.001). In cooler months, no preference to any height level (F=1.67, df=6, p=0.2) was observed. Proportion of females sand flies with Sella's classification of abdominal stages, namely full-fed, half-gravid and gravid females did not vary significantly (t=1.98, p=0.13827) indoors, confirming their endophilic behaviour. Risk of CL transmission in these tribal settlements is discussed.

Seven novel single nucleotide polymorphisms identified within river buffalo (Bubalus bubalis) lactoferrin gene.

The present study aimed at identifying single-nucleotide polymorphic (SNP) sites in different coding and non-coding regions of lactoferrin gene in Indian riverine buffaloes. A total of 102 animals from six different river buffalo breeds were screened at six bubaline lactoferrin gene loci. Single-strand conformation polymorphism (SSCP) analysis revealed monomorphic patterns at three loci LtfE2, LtfE11, and LtfE14 while a total of eight distinct patterns were observed in the other three loci viz. LtfE5, LtfE10, and LtfE16 which correspond to respective exons and their flanking regions. Sequence analysis of different SSCP variants revealed the presence of two SNP sites within the coding (exon 16) region and five SNP sites in flanking non-coding regions (intron 4 and intron 9). Both SNPs within exon 16 were found to be synonymous. The SNPs and haplotypes identified in the present study could serve as potential markers for association with susceptibility/resistance to mastitis in buffaloes.

Exonic splicing enhancers contribute to the use of both 3' and 5' splice site usage of rat beta-tropomyosin pre-mRNA.

The rat beta-tropomyosin gene encodes two tissue-specific isoforms that contain the internal, mutually exclusive exons 6 (nonmuscle/smooth muscle) and 7 (skeletal muscle). We previously demonstrated that the 3' splice site of exon 6 can be activated by introducing a 9-nt polyuridine tract at its 3' splice site, or by strengthening the 5' splice site to a U1 consensus binding site, or by joining exon 6 to the downstream common exon 8. Examination of sequences within exons 6 and 8 revealed the presence of two purine-rich motifs in exon 6 and three purine-rich motifs in exon 8 that could potentially represent exonic splicing enhancers (ESEs). In this report we carried out substitution mutagenesis of these elements and show that some of them play a critical role in the splice site usage of exon 6 in vitro and in vivo. Using UV crosslinking, we have identified SF2/ASF as one of the cellular factors that binds to these motifs. Furthermore, we show that substrates that have mutated ESEs are blocked prior to A-complex formation, supporting a role for SF2/ASF binding to the ESEs during the commitment step in splicing. Using pre-mRNA substrates containing exons 5 through 8, we show that the ESEs within exon 6 also play a role in cooperation between the 3' and 5' splice sites flanking this exon. The splicing of exon 6 to 8 (i.e., 5' splice site usage of exon 6) was enhanced with pre-mRNAs containing either the polyuridine tract in the 3' splice site or consensus sequence in the 5' splice site around exon 6. We show that the ESEs in exon 6 are required for this effect. However, the ESEs are not required when both the polyuridine and consensus splice site sequences around exon 6 were present in the same pre-mRNA. These results support and extend the exon-definition hypothesis and demonstrate that sequences at the 3' splice site can facilitate use of a downstream 5' splice site. In addition, the data support the hypothesis that ESEs can compensate for weak splice sites, such as those found in alternatively spliced exons, thereby providing a target for regulation.

Monocyte derived IL 10 and PGE2 are associated with the absence of Th 1 cells and in vitro T cell suppression in lepromatous leprosy.

Our previous studies had shown that the clinicopathological spectrum in leprosy was associated with discrete T cell subsets in circulation, with tuberculoid patients having antigen-induced Th 1, whereas lepromatous leprosy patients with antigen-specific T cell anergy possessed Th 2 cells. The present study shows that infected monocytes from lepromatous but not tuberculoid leprosy patients released soluble factors (MoF(s)) containing IL-10 and PGE2 which inhibited M. leprae induced in vitro lymphoproliferation of previously sensitised healthy or tuberculoid leprosy subjects. A strong negative correlation was observed between adherent cell derived IL-10 and IL-2 at the level of both the product and cytokine mRNA. Moreover, anti-IL-10 antibodies and indomethacin partially reversed the suppressor effects of MoF(s). Taken together these studies indicate that infected monocytes contribute to the development of T cell anergy by releasing factors that affect regulatory cytokines and T cell subset differentiation in lepromatous leprosy.