EVALUATING THE IMPACT OF SOLID MICRONEEDLES ON THE TRANSDERMAL DRUG DELIVERY SYSTEM FOR Ɣ-ORYZANOL

Objective: This study's goals were to develop a minimally invasive array of biocompatible polymeric solid microneedles and formulate a transdermal patch of drug Ɣ-Oryzanol as per poke and patch technology. Methods: Scanning electron microscopy was used to analyse the morphology of the solid microneedle arrays, which were created using a stereolithography (SLA) printer with high-resolution capabilities (25 and 140 microns for the z and x axes, respectively). Transdermal Patches of Ɣ-Oryzanol were formulated and evaluated for various characterization parameters. Further, the produced microneedle-transdermal drug delivery system of Ɣ-Oryzanol was examined for microneedle insertion skin and permeation of the drug across the porcine skin. Results: Solid microneedle arrays were manufactured using biocompatible Class I Dental SG resin having dimensions of 600 µm height and 300 µm width with tip diameters of 30 µm and 1.85 mm interspacing (Distance from tip to tip) and they were strong enough to penetrate porcine skin to a depth of 381.356 µm crossing the stratum corneum layer without causing any structural changes. Transdermal patches containing Ɣ-Oryzanol were formulated using different ratios of HPMC: Eudragit E-100. Good, consistent, and transparent films were formulated when the thickness of the film ranges between 0.516±0.25-0.628±0.21 mm, average weights ranged from 168.23±2.61to171.22±1.25(10/cm2), folding endurance ranged in between 10 folds to 12 folds for all the formulations with tensile strength lie between the 0.365 kg/mm2 to 0.465 kg/mm2. All the formulations showed good drug content between 99.3±0.06%-90.4±1.64% with 100% flat surfaces. Moisture content was found in the range of 2.012±0.013 to 4.213±0.031. Drug permeation studies reveal that compound Ɣ-Oryzanol transdermal patches didn’t show significant permeation across porcine skin (4.802.25 g/cm2) without piercing with microneedles while after poking skin using microneedles (74.502.35 g/cm2) drug showed good penetration properties. It was found that the amount of drug delivered increased to 44.251.57 g/cm2 at 2 min, which was 14.502.35 g/cm2 at 1 min to 4 min 74.502.35 g/cm2. Conclusion: Successful preparation of the Microneedle-Transdermal drug delivery system of Ɣ-Oryzanol and their evaluation indicated that the quality and consistency of the formulated preparation were excellent. With advantages in terms of lowered dose frequency, better patient compliance, and bioavailability, this may find use in the therapeutic field

IN VITRO ABSORPTION STUDY OF CARBAMAZEPINE SOLID DISPERSION USING EVERTED GUT SAC METHOD

The oral Bioavailability of BCS (Bio Pharmaceutical Classification System) class II drug with poor solubility and reasonable permeability is limited by drug dissolution. In order to improve the aqueous solubility of the drug and dissolution of thedrug, the solid dispersion was prepared and evaluated for its absorption in intestine using modified everted gut sac method. The solid dispersion of carbamezepine (CBZ) was prepared using polaxomer and guargum by kneading method. The CBZ and CBZSD (Solid Disposisi) shows 2.329% and 3.948% drug absorption, respectively. The data show that solid dispersion increase the absorption of the CBZ in CBZ-SD is more than 70% in comparison to pure CBZ. The increase in CBZ solubility of the SD could be attributed to several factors such as improved wettability, local solubilization, drug particle size reduction and crystalline or, interstitial solid solution reduction. Key words: Everted gut sac method, solid dispersion, absorption, solubilit

Study on the Phytochemical, Antioxidant and In Vitro Anticancer activity on root extract of Simarouba glauca DC

Plant made medicine plays main role in drug formulation and synthesis with moderate or no side effects. Simarouba glauca is exotic to India and known for its phytomedicine property belonging to the Simaroubaceae family prominent for its leaves arrangement and is called “Lakshmi taru” or “Paradise tree”. The goal of this study was to present the preliminary phytochemicals, antioxidants, and cytotoxicity mechanism of S. glauca root extracts in terms of apoptosis in cell cycle. Phytochemical, qualitative, and antioxidant assay analyses were performed with the standard protocol using HPTLC and cytotoxicity studies were analysed using a flow-cytometer. The Phytochemical screening revealed the presence of saponins, flavonoids, steroids, and triterpenoids. Furthermore, the extracts demonstrated potent antioxidant activity and could serve as a lead source of natural antioxidants. In a concentration-dependent manner, anticancer study demonstrated cytotoxicity against the A549 cell line by stopping the cells at the S-phase. The main investigation reveals the antiproliferative properties of S. glauca methanolic root extract. Identification of cytotoxic compounds and their mode of action require further in-depth research. The outcome of our investigation offers compelling evidence that S. glauca methanolic root extract can be used as an effective ethnomedicinal agent with the ability to treat human disorders

HIVsirDB: A Database of HIV Inhibiting siRNAs

Human immunodeficiency virus (HIV) is responsible for millions of deaths every year. The current treatment involves the use of multiple antiretroviral agents that may harm patients due to their toxic nature. RNA interference (RNAi) is a potent candidate for the future treatment of HIV, uses short interfering RNA (siRNA/shRNA) for silencing HIV genes. In this study, attempts have been made to create a database HIVsirDB of siRNAs responsible for silencing HIV genes.HIVsirDB is a manually curated database of HIV inhibiting siRNAs that provides comprehensive information about each siRNA or shRNA. Information was collected and compiled from literature and public resources. This database contains around 750 siRNAs that includes 75 partially complementary siRNAs differing by one or more bases with the target sites and over 100 escape mutant sequences. HIVsirDB structure contains sixteen fields including siRNA sequence, HIV strain, targeted genome region, efficacy and conservation of target sequences. In order to facilitate user, many tools have been integrated in this database that includes; i) siRNAmap for mapping siRNAs on target sequence, ii) HIVsirblast for BLAST search against database, iii) siRNAalign for aligning siRNAs.

Przewlekła poreumatyczna artropatia — zespół Jaccouda u 21-letniego chorego

Jaccoud’s arthropathy (JA) is a condition characterised clinically by ‘reversible’ joint deformities resulting from soft tissue abnormalities, rather than the destruction of bone joints. Although rare, it has been described in patients with chronic rheumatic heart disease. It typically involves metacarpophalangeal and metatarsophalangeal joints. JA has also been described in systemic lupus erythematosus (SLE), infections and neoplasia. Here, we report the case of a 21 year-old male who had presented with a 16-year history of polyarthralgia, deformities of the hands for the last four years, and a three-year history of exertional palpitation and dyspnoea. Physical examination revealed peripheral signs of aortic run off, with hyperdynamic left ventricle and chronic severe aortic and mitral regurgitation. On examination, gross deformities were noted in both hands in the form of ulnar deviation, swan neck, and ‘z’-thumb, involving interphalangeal joints, metacarpophalangeal and first carpometacarpal joints which were partially correctable. Laboratory investigations ruled out rheumatoid arthritis, SLE, and acute rheumatic fever. Chest X-ray, electrocardiogram and two-dimensional echocardiography and Doppler study were consistent with chronic severe aortic regurgitation and severe mitral regurgitation. Radiographs of both hands showed posterior subluxation of the carpometacarpal joints, which were completely correctable. JA involving a carpometacarpal joint is exceedingly rare.Artropatia (zespół) Jaccouda jest schorzeniem charakteryzującym się klinicznie „odwracalnymi” deformacjami stawówspowodowanymi zaburzeniami tkanek miękkich, a nie niszczeniem kostnych elementów stawów. Chociaż artropatia tawystępuje rzadko, to opisywano przypadki JA u chorych z przewlekłą chorobą reumatyczną serca. Zazwyczaj obejmujestawy śródręczno-paliczkowe i śródstopno-paliczkowe. Zespół ten obserwowano również w toczniu rumieniowatymukładowym (SLE), zakażeniach i chorobach nowotworowych. W niniejszej pracy przedstawiono przypadek 21-letniegochorego, u którego od 16 lat występowały bóle wielu stawów, od 4 lat — deformacje rąk i od 3 lat — wysiłkowe kołatanieserca i duszność. W badaniu przedmiotowym stwierdzono obwodowe objawy wstecznego przepływu krwi w aorcie orazhiperdynamiczną czynność lewej komory i przewlekłą ciężką niedomykalność aortalną i mitralną. W badaniu wzrokowymwyraźnie widoczne były deformacje obu rąk w postaci odchylenia łokciowego, deformacji typu „łabędzia szyja” i kciukaw kształcie litery „z”. Na podstawie wyników badań laboratoryjnych wykluczono reumatoidalne zapalenie stawów, SLEi ostrą gorączkę reumatyczną. Wyniki badań obrazowych — badania radiologicznego (RTG) klatki piersiowej, elektrokardiografii,echokardiografii dwuwymiarowej i badania doplerowskiego — wskazywały na przewlekłą ciężką niedomykalnośćaortalną i mitralną. W obrazie RTG obu rąk uwidoczniono podwichnięcia ku tyłowi w stawach nadgarstkowo-śródręcznych,które można było całkowicie skorygować. Artropatia Jaccouda obejmująca staw nadgarstkowo-śródręczny jestobserwowana wyjątkowo rzadko

Protective Role of Ashwagandha Leaf Extract and Its Component Withanone on Scopolamine-Induced Changes in the Brain and Brain-Derived Cells

BACKGROUND:Scopolamine is a well-known cholinergic antagonist that causes amnesia in human and animal models. Scopolamine-induced amnesia in rodent models has been widely used to understand the molecular, biochemical, behavioral changes, and to delineate therapeutic targets of memory impairment. Although this has been linked to the decrease in central cholinergic neuronal activity following the blockade of muscarinic receptors, the underlying molecular and cellular mechanism(s) particularly the effect on neuroplasticity remains elusive. In the present study, we have investigated (i) the effects of scopolamine on the molecules involved in neuronal and glial plasticity both in vivo and in vitro and (ii) their recovery by alcoholic extract of Ashwagandha leaves (i-Extract). METHODOLOGY/PRINCIPAL FINDINGS:As a drug model, scopolamine hydrobromide was administered intraperitoneally to mice and its effect on the brain function was determined by molecular analyses. The results showed that the scopolamine caused downregulation of the expression of BDNF and GFAP in dose and time dependent manner, and these effects were markedly attenuated in response to i-Extract treatment. Similar to our observations in animal model system, we found that the scopolamine induced cytotoxicity in IMR32 neuronal and C6 glioma cells. It was associated with downregulation of neuronal cell markers NF-H, MAP2, PSD-95, GAP-43 and glial cell marker GFAP and with upregulation of DNA damage--γH2AX and oxidative stress--ROS markers. Furthermore, these molecules showed recovery when cells were treated with i-Extract or its purified component, withanone. CONCLUSION:Our study suggested that besides cholinergic blockade, scopolamine-induced memory loss may be associated with oxidative stress and Ashwagandha i-Extract, and withanone may serve as potential preventive and therapeutic agents for neurodegenerative disorders and hence warrant further molecular analyses

IBEX:A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues

The diverse composition of mammalian tissues poses challenges for understanding the cell–cell interactions required for organ homeostasis and how spatial relationships are perturbed during disease. Existing methods such as single-cell genomics, lacking a spatial context, and traditional immunofluorescence, capturing only two to six molecular features, cannot resolve these issues. Imaging technologies have been developed to address these problems, but each possesses limitations that constrain widespread use. Here we report a method that overcomes major impediments to highly multiplex tissue imaging. “Iterative bleaching extends multiplexity” (IBEX) uses an iterative staining and chemical bleaching method to enable high-resolution imaging of >65 parameters in the same tissue section without physical degradation. IBEX can be employed with various types of conventional microscopes and permits use of both commercially available and user-generated antibodies in an “open” system to allow easy adjustment of staining panels based on ongoing marker discovery efforts. We show how IBEX can also be used with amplified staining methods for imaging strongly fixed tissues with limited epitope retention and with oligonucleotide-based staining, allowing potential cross-referencing between flow cytometry, cellular indexing of transcriptomes and epitopes by sequencing, and IBEX analysis of the same tissue. To facilitate data processing, we provide an open-source platform for automated registration of iterative images. IBEX thus represents a technology that can be rapidly integrated into most current laboratory workflows to achieve high-content imaging to reveal the complex cellular landscape of diverse organs and tissues

Chlorhexidine for facility-based umbilical cord care: EN-BIRTH multi-country validation study.

BACKGROUND: Umbilical cord hygiene prevents sepsis, a leading cause of neonatal mortality. The World Health Organization recommends 7.1% chlorhexidine digluconate (CHX) application to the umbilicus after home birth in high mortality contexts. In Bangladesh and Nepal, national policies recommend CHX use for all facility births. Population-based household surveys include optional questions on CHX use, but indicator validation studies are lacking. The Every Newborn Birth Indicators Research Tracking in Hospitals (EN-BIRTH) was an observational study assessing measurement validity for maternal and newborn indicators. This paper reports results regarding CHX. METHODS: The EN-BIRTH study (July 2017-July 2018) included three public hospitals in Bangladesh and Nepal where CHX cord application is routine. Clinical-observers collected tablet-based, time-stamped data regarding cord care during admission to labour and delivery wards as the gold standard to assess accuracy of women's report at exit survey, and of routine-register data. We calculated validity ratios and individual-level validation metrics; analysed coverage, quality and measurement gaps. We conducted qualitative interviews to assess barriers and enablers to routine register-recording. RESULTS: Umbilical cord care was observed for 12,379 live births. Observer-assessed CHX coverage was very high at 89.3-99.4% in all 3 hospitals, although slightly lower after caesarean births in Azimpur (86.8%), Bangladesh. Exit survey-reported coverage (0.4-45.9%) underestimated the observed coverage with substantial "don't know" responses (55.5-79.4%). Survey-reported validity ratios were all poor (0.01 to 0.38). Register-recorded coverage in the specific column in Bangladesh was underestimated by 0.2% in Kushtia but overestimated by 9.0% in Azimpur. Register-recorded validity ratios were good (0.9 to 1.1) in Bangladesh, and poor (0.8) in Nepal. The non-specific register column in Pokhara, Nepal substantially underestimated coverage (20.7%). CONCLUSIONS: Exit survey-report highly underestimated observed CHX coverage in all three hospitals. Routine register-recorded coverage was closer to observer-assessed coverage than survey reports in all hospitals, including for caesarean births, and was more accurately captured in hospitals with a specific register column. Inclusion of CHX cord care into registers, and tallied into health management information system platforms, is justified in countries with national policies for facility-based use, but requires implementation research to assess register design and data flow within health information systems

Réseaux de signalisation intégrés dans l'immunité innée chez C. elegans

C. elegans est infecté par divers agents pathogènes ; bactéries, champignons et virus. Lors d'une infection fongique, C. elegans surexprime de nombreux gènes codant pour des peptides antimicrobiens (AMP). Le principal objectif de ma thèse était de construire un réseau de régulation génique intégré représentant l'induction de ces gènes AMP pendant l'infection. Pour trouver les principales composantes du réseau de régulation, via un criblage ARNi du génome entier (Zugasti et al 2016), nous avons identifié 278 clones Nipi (pour "Absence d'induction de peptides antimicrobiens après infection") qui abrogent l’induction d’AMP. En utilisant "CloneMapper" (Thakur et al. 2014), nous avons identifié 338 gènes cibles pour ces clones. Nous avons montré que les voies de MAPK sont au cœur de l'induction des AMP. Parmi les 50 gènes arbitrairement sélectionnés et surexprimant les AMP, nous en avons validé 48 en utilisant Fluidigm. Pour attribuer des fonctions aux gènes identifiés dans ces études à haut débit, nous avons développé un outil d'enrichissement fonctionnel pour la communauté C.elegans (MS en préparation). Nous avons utilisé cet outil pour analyser les cibles du criblage ARNi sur le génome entier et sur d'autres bases de données concernant divers agents pathogènes. Nous avons fait une analyse de l'enrichissement fonctionnel des cibles ChIPseq de CEBP-1, un facteur de transcription lié à la régulation de la réponse immunitaire innée (Kim et al, Soumis). Enfin, pour mieux comprendre l'interaction entre l'hôte et le pathogène, nous avons séquencé, assemblé, annoté et analysé le génome de D. coniospora. Nous avons identifié plusieurs facteurs de virulence potentiels dans ce génome.C. elegans is infected by diverse pathogens, including bacteria, fungi and viruses. Upon fungal infection, C. elegans up-regulates the expression of many antimicrobial peptide (AMP) genes. The main aim of my thesis was to build an integrated gene regulatory network representing the induction of these AMP genes upon infection. To find the main/backbone components of the regulatory network, through a genome-wide RNAi screen (Zugasti et al. 2016), we identified 278 Nipi (for “no induction of antimicrobial peptides after infection”) clones that abrogate AMP induction. Using “CloneMapper” (Thakur et al. 2014), we identified 338 target genes for these clones. We showed that MAPK pathways are central to the induction of AMPs. We also characterized the transcriptional changes provoked by infection using RNA-sequencing and identified more than 300 genes that are dynamically up-regulated after infection, including 13 AMPs. We validated 48 (96%) of 50 arbitrary selected up-regulated genes using Fluidigm. To assign functions to genes identified in these high-throughput studies, we developed a functional enrichment tool for C.elegans community (MS in preparation). We used this tool to analyse the genome-wide RNAi screen targets and other pathogen-related datasets. We did functional enrichment analysis of ChIPseq targets of CEBP-1, TF linked to the regulation of the innate immune response (Kim et al., submitted). Finally, to understand better the interaction between host and pathogen, we sequenced, assembled, annotated and analysed the D. coniospora genome (Lebrigand et al. 2016). We identified various potential virulence factors in the fungal genome