Experimental infection and pathology of two highly pathogenic avian influenza H5N1 viruses isolated from crow and chicken in house crows (Corvus splendens).

We investigated the experimental infection of two highly pathogenic avian influenza H5N1 viruses isolated from crow (A/crow/Assam/142119/2008) and chicken (A/chicken/Sikkim/151466/2009) in house crows (Corvus splendens). Both viruses caused infection in crows, where four out of six and three out of six crows succumbed to H5N1 infection within 11 days post challenge by crow and chicken viruses, respectively. The major clinical signs in crows were wing paralysis, circling and torticollis. The virus shedding detected from swabs was not persistent in both crow nor chicken viruses. Both viruses were isolated more frequently from oral swabs than from cloacal swabs. Both virus strains were isolated from brain, lungs, heart, liver, pancreas, spleen, large intestines of crows that succumbed to H5N1 infection. The surviving birds seroconverted in response to H5N1 virus infection. Microscopically, both viruses caused coagulative necrosis in pancreas and kidneys. Brain showed gliosis and neuronal degeneration. This experimental study highlights that crows could be infected with H5N1 viruses from different hosts with minor differences in pathogenicity. Therefore, it is imperative to carry out surveillance of highly pathogenic avian influenza H5N1 virus in synanthropic birds along with biosecurity measures to mitigate the H5N1 spread in poultry population. Keywords: chicken virus; crow virus; highly pathogenic avian influenza; house crows.

Norfloxacin salts of carboxylic acids curtail planktonic and biofilm mode of growth in ESKAPE pathogens.

AIMS: To enhance the antimicrobial and antibiofilm activity of norfloxacin against the planktonic and biofilm mode of growth in ESKAPE pathogens using chemically modified norfloxacin salts. METHODS AND RESULTS: Antimicrobial testing, synergy testing and time-kill curve analysis were performed to evaluate antibacterial effect of norfloxacin carboxylic acid salts against ESKAPE pathogens. In vivo efficacy to reduce bacterial bioburden was evaluated in zebrafish infection model. Crystal violet assay and live-dead staining were performed to discern antibiofilm effect. Membrane permeability, integrity and molecular docking studies were carried out to ascertain the mechanism of action. The carboxylic acid salts, relative to parent molecule norfloxacin, displayed two- to fourfold reduction in minimum inhibitory concentration against Staphylococcus aureus and Pseudomonas aeruginosa, in addition to displaying potent bacteriostatic effect against certain members of ESKAPE pathogens. In vivo treatments revealed that norfloxacin tartrate (SRIN2) reduced MRSA bioburden by greater than 1 log fold relative to parent molecule in the muscle tissue. In silico docking with gyrA of S. aureus showed increased affinity of SRIN2 towards DNA gyrase. The enhanced antibacterial effect of norfloxacin salts could be partially accounted by altered membrane permeability in S. aureus and perturbed membrane integrity in P. aeruginosa. Antibiofilm studies revealed that SRIN2 (norfloxacin tartrate) and SRIN3 (norfloxacin benzoate) exerted potent antibiofilm effect particularly against Gram-negative ESKAPE pathogens. The impaired colonization of both S. aureus and P. aeruginosa due to improved norfloxacin salts was further supported by live-dead imaging. CONCLUSION: Norfloxacin carboxylic acid salts can act as potential alternatives in terms of drug resensitization and reuse. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study shows that carboxylic acid salts of norfloxacin could be effectively employed to treat both planktonic- and biofilm-based infections caused by select members of ESKAPE pathogens.

Amantadine resistance among highly pathogenic avian influenza viruses (H5N1) isolated from India.

Emergence of antiviral resistance among H5N1 avian influenza viruses is the major challenge in the control of pandemic influenza. Matrix 2 (M2) inhibitors (amantadine and rimantadine) and neuraminidase inhibitors (oseltamivir and zanamivir) are the two classes of antiviral agents that are specifically active against influenza viruses and are used for both treatment and prophylaxis of influenza infections. Amantadine targets the M2 ion channel of influenza A virus and interrupts virus life cycle through blockade of hydrogen ion influx. This prevents uncoating of the virus in infected host cells which impedes the release of ribonucleoprotein required for transcription and replication of virion in the nucleus. The present study was carried out to review the status of amantadine resistance in H5N1 viruses isolated from India and to study their replicative capability. Results of the study revealed resistance to amantadine in antiviral assay among four H5N1 viruses out of which two viruses had Serine 31 Asparagine (AGT-AAT i.e., S31N) mutation and two had Valine 27 Alanine (GTT-GCT i.e., V27A) mutation. The four resistant viruses not only exhibited significant difference in effective concentration 50% (EC50) values of amantadine hydrochloride from that of susceptible viruses (P < 0.0001) but also showed significant difference between two different types (S31N and V27A) of mutant viruses (P < 0.05). Resistance to amantadine could also be demonstrated in a simple HA test after replication of the viruses in MDCK cells in presence of amantadine. The study identifies the correlation between in vitro antiviral assay and presence of established molecular markers of resistance, the retention of replicative capacity in the presence of amantadine hydrochloride by the resistant viruses and the emergence of resistant mutations against amantadine among avian influenza viruses (H5N1) without selective drug pressure.

Expression and purification of recombinant H5HA1 protein of H5N1 avian influenza virus in E. coli and its application in indirect ELISA.

The PCR amplified HA1 fragment of H5N1 (H5HA1) avian influenza virus (AIV) hemagglutinin gene was cloned into pET28a (+) expression vector and expressed in Rosetta Blue (DE3) pLysS cells. The recombinant H5HA1 (rH5HA1) protein purified by passive gel elution after SDS-PAGE of the inclusion bodies reacted specifically with H5N1 serum in Western blot analysis. A subtype specific indirect enzyme linked immunosorbent assay (iELISA) using the rH5HA1 protein as the coating antigen was developed for detecting antibodies to H5 subtype of AIV. The assay had 89.04% sensitivity and 95.95% specificity when compared with haemagglutination inhibition test. The Kappa value of 0.842 indicated a perfect agreement between the tests. The iELISA developed can be used for serosurveillance of avian influenza in chickens.

Eosinophilic annular erythema: successful response to ultraviolet B therapy.

Eosinophilic annular erythema (EAE) is a rare and relatively newly described eosinophil-rich dermatosis. Debate still exists as to whether it represent a subtype of Well syndrome or a separate disease entity. We report an 8-year-old boy with a 4-year history of recurrent, asymptomatic annular lesions, which were diagnosed after clincopathological correlation as EAE. This condition usually runs a relapsing and remitting course with resistance to multiple treatments. Prednisolone and hydroxychloroquine have been reported as successful but the response to these was limited in this case. Complete resolution occurred after treatment with ultraviolet B (UVB) therapy. To our knowledge, this is the first report of a favourable response of EAE to such therapy.

Pathology of a H5N1, highly pathogenic avian influenza virus, in two Indian native chicken breeds and a synthetic broiler line.

In this study, susceptibility to H5N1 virus infection was studied in two Indian native chicken breeds viz. Kadaknath and Aseel (Peela) and an Indian synthetic broiler strain (Synthetic dam line (SDL-IC). Fifty birds from each genetic group were infected intra-nasally with 1000 EID50 of a highly pathogenic avian influenza virus (HPAIV) strain A/chicken/Navapur/India/7972/ 06 (H5N1) and observed for a period of 10 days. Significant differences in severity of clinical signs, gross lesions and time for onset of symptoms were observed. The overall severity of clinical signs and gross lesions was less in SDL-IC broilers as compared to the other two genetic groups. The mortality percentages were 100, 98 and 92% with Mean Death Time (MDT) of 3.12, 5.92 and 6.96 days, respectively for the two native breeds Kadaknath and Aseel (Peela), the and SDL-IC broiler strain. Comparison of histological lesions revealed differences in disease progression among the genetic groups. Vascular lesions such as disseminated intravascular coagulopathy (DIC) were predominant on 3 days post infection (dpi) in Kadaknath, and on 5 and 6 dpi in Aseel (Peela) and SDL-IC broiler. The mean log2 HA titres of the re-isolated virus from various organs of H5N1 AIV infected birds of the three genetic groups ranged from 2.32 (lung, trachea and bursa) to 5.04 (spleen) in Kadaknath; 2.32 (lung) to 6.68 (brain) in Aseel (Peela); and 2.06 (liver) to 7.01 (lungs and kidney) in SDL-IC broiler. These results suggest that the susceptibility to H5N1 highly pathogenic avian influenza virus infection differed among the three breeds; Kadaknath being highest followed by Aseel (Peela) and synthetic SDL-IC broiler. This is possibly the first report on the differences in the susceptibility of the India native breeds to H5N1 virus infection and its severity.

Influence of thermal processing on the volatile constituents of muskmelon puree.

Muskmelon (Cucumis melo L) is an important tropical fruit cultivated widely in different parts of India. Fresh muskmelon has a delicate but characteristic flavor rendering the fruit with highly acceptable flavor. Processing and preservation of muskmelon puree requires thermal processing, which affects the volatile constituents. It is imperative to understand the flavor changes during thermal processing which would affect the quality of the processed and packed muskmelon puree. Muskmelon puree was subjected to different methods of thermal processing viz., heating, canning and packing in retort pouches and the volatile constituents were analyzed. Gas chromatography-mass spectrometry (GC-MS) indicated the presence of more than 49 volatile components in the muskmelon puree samples. Major volatile components identified using GC-MS analysis showed the presence of esters (27.29 %), aldehydes (18.57 %), Heterocyclic compounds (16.63 %), aliphatic alcohols (11.72 %), phenolic compounds (6.03 %) and sesquiterpenes (0.25 %) in the fresh samples. Aldehydes decreased and ester content increased in thermally processed muskmelon puree packed in cans and retort pouches. Aliphatic alcohols, Heterocyclic compounds and phenolic compounds decreased in puree processed in tin containers and retort pouches.

2,5-Di-methyl-phenyl quinoline-2-carboxy-l-ate.

In the title compound, C18H15NO2, the dihedral angle between the mean planes of the quinoline ring system and the phenyl ring is 78.8 (1)°. The mean plane of the carboxyl-ate group is twisted from the mean planes of the quinoline ring system and phenyl ring by 1.5 (9) and 77.6 (4)°, respectively. In the crystal, mol-ecules are linked by weak C-H⋯O inter-actions, generating C(8) chains along [001]. Weak π-π stacking inter-actions are also observed [centroid-centroid separation = 3.6238 (12) Å].

2-Isopropyl-5-methyl-cyclo-hexyl quinoline-2-carboxyl-ate.

In the title compound, C20H25NO2, the cyclo-hexyl ring adopts a slightly disordered chair conformation. The dihedral angle between the mean planes of the quinoline ring and the carboxyl-ate group is 22.2 (6)°. In the crystal, weak C-H⋯N inter-actions make chains along [010].

Incidence and Management of Pharyngo Cutaneous Fistula Following Total Laryngectomy - A Single Institutional 10 Years Experience.

Pharyngocutaneous fistula (PCF) is a common postoperative complication in patients undergoing laryngectomy. Many studies have focused on the predisposing factors and their treatment, but there is still controversy on how to identify high-risk patients.PCF is associated with delay in adjuvant treatment, prolonged hospital stay, and the requirement for additional surgical procedures. The increased incidence of post-laryngectomy PCF in the modern era of organ preservation therapy has driven considerable efforts to develop techniques to avoid and treat this complication. The study aims to analyze the incidence, predisposing factors, management, and outcome of PCF in post laryngectomy patients. We reviewed retrospectively the medical records of 72 patients who underwent total laryngectomy during the period from 2013 to 2022 at our institute. Details regarding age, gender, tumor site, comorbidities, radiotherapy, chemotherapy, duration of nasogastric tube feeding, the incidence of PCF and its management, and length of hospital stay. In our analysis of 72 patients, with 38 undergoing upfront laryngectomy and 34 salvage laryngectomy post-chemoradiation, we observed a 23% incidence of Pharyngocutaneous Fistula (PCF). Our univariate analysis revealed that low pre-op albumin levels and advanced stage significantly increased the risk of PCF. Specifically, PCF occurred in 16% of upfront laryngectomy patients and increased to 32% in salvage laryngectomy patients, though there was no significant association with pre-operative radiotherapy (RT).Among PCF cases, those from upfront laryngectomy had a mean onset time of 12 days, while salvage laryngectomy cases had a mean onset time of 8 days. The majority of PCF cases (64%) were managed conservatively, with 11% resolving through secondary suturing and 25% requiring surgical repair using local flaps. Notably, 80% of upfront laryngectomy PCF cases were managed conservatively, compared to 54% in salvage laryngectomy cases.The average hospitalization time for PCF patients was significantly longer at 35.6 days, compared to 12 days for non-PCF patients. Importantly, there were no fatal complications related to PCF in either group. Our study found that Pharyngocutaneous Fistula (PCF) occurs in both irradiated and non-irradiated patients, with T staging and pre-op albumin levels being significant risk factors. While PCF in non-irradiated patients can be treated conservatively, those in chemoradiation patients often require surgical intervention. Postoperative nutritional support like Feeding jejunostomy or Peg tube insertion aids in patient recovery from PCF.

Prevalence and risk factor for H9N2 avian influenza virus in poultry retail shops of Madhya Pradesh.

H9N2 avian Influenza virus subtype is highly neglected but have the potential to emerge as a next pandemic influenza virus, by either itself evolution or through the donation of genes to other subtype. So to understand the extent of H9N2 virus prevalence and associated risk factors in poultry of retail shops and their surrounding environment a cross sectional study was carried out. A total of 500 poultry tissue and 700 environmental samples were collected from 20 district of Madhya Pradesh. Virus isolation was carried out in egg inoculation and harvested allantoic fluid was tested for HA and further molecular confirmation of subtypes by RT-PCR using H9 specific primers. Prevalence was calculated and positive samples were statistically associated with observed risk factors using univariate and multivariate logistic regression analysis. A total of 9.4% and 9.7% prevalence in tissue samples and environmental samples has been reported respectively and out of 20 districts 10 (50%) were found positive for the virus. Out of 21 studied risk factors only two risk factors named as "keeping total number birds slaughtered per day" and "procuring birds from wholesaler" were found significantly associated with the H9N2 positivity in multivariate logistic regression analysis. This high level of H9N2 positivity in birds with no clinical manifestations providing a great opportunity for avian influenza virus for amplification, co-infection in other animals like dogs, cats, pigs and in human through genetic re-assortment that may lead to emergence of a novel influenza virus with high zoonotic potential. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-024-00865-y.

IL-17 induces expression of vascular cell adhesion molecule through signalling pathway of NF-κB, but not Akt1 and TAK1 in vascular smooth muscle cells.

Interleukin-17 (IL-17) plays an important role in several autoimmune diseases. IL-17 can induce the expression of vascular cell adhesion molecule (VCAM-1) in aortic vascular smooth muscle cells (SMCs), which is important for the development of atherosclerosis. However, the signalling pathway of IL-17-induced VCAM-1 expression remains unclear. In this study, we reported that IL-17-induced expression of VCAM-1 in SMCs is dependent on NF-κB, but independent of Akt1 and TAK1. This is because knocking down Akt1 or TAK1 by siRNA did not reduce IL-17-induced activation of NF-κB and expression of VCAM-1, whereas knocking down NF-κB by siRNA markedly inhibited IL-17-mediated upregulation of VCAM-1 expression. In addition, IL-17-induced expression of VCAM-1 is partially dependent on activation of ERK1/2. Therefore, these signalling pathways of IL-17-mediated upregulation of VCAM-1 expression might be therapeutic targets for treatment of IL-17-mediated inflammation.

STM imaging, spectroscopy and manipulation of a self-assembled PTCDI monolayer on epitaxial graphene.

Scanning Tunneling Microscopy (STM), Scanning Tunneling Spectroscopy (STS), and manipulation studies were performed on an ordered self-assembled monolayer (SAM) of N,N'-bis(1-hexylheptyl)perylene-3,4:9,10-bis(dicarboximide) molecules on epitaxial graphene on hexagonal silicon carbide - SiC(0001). Four novel aspects of the molecular SAM on graphene are presented. Molecules adsorb in both armchair and zig-zag configurations, giving rise to six orientations of the molecular layer with respect to the underlying substrate. The interaction between the molecules and the graphene surface shifts the LUMO towards the Fermi level, inducing a charge transfer and the opening of a band gap in the graphene, with the LUMO inside. This decouples the LUMO from the surface rendering it invisible in the dI/dV spectroscopy. The HOMO only becomes visible at short tip-surface distances, as its energy lies within the band gap of the SiC substrate. Finally, the observed molecular defects are very particular, being composed exclusively of molecular dimers. These molecular dimers have a stronger interaction with the graphene than other molecules.

4-Chloro-phenyl quinoline-2-carboxyl-ate.

In the title compound, C16H10ClNO2, the dihedral angle between the quinoline ring system and the benzene ring is 14.7 (5)°. The carboxyl-ate group is twisted from the mean planes of the quinoline ring system and the benzene ring by 17.7 (5) and 32.1 (4)°, respectively. In the crystal, inversion dimers are formed with the molecules linked by pairs of weak C-H⋯O inter-actions arising from an activated aromatic C atom adjacent to the C-Cl bond, generating R 2 (2)(14) loops.

4-Chloro-3-methyl-phenyl quinoline-2-carboxyl-ate.

In the title compound, C17H12ClNO2, the dihedral angle between the mean planes of the quinoline ring system and the benzene ring is 68.7 (7)°. The mean plane of the carboxyl-ate group is twisted from the latter planes by 14.0 (1) and 80.2 (4)°, respectively. In the crystal, weak C-H⋯O inter-actions are observed, forming chains along [001]. In addition, π-π stacking inter-actions [centroid-centroid distances = 3.8343 (13) and 3.7372 (13)Å] occur. No classical hydrogen bonds were observed.

3,4-Di-methyl-phenyl quinoline-2-carboxyl-ate.

In the title compound, C18H15NO2, the dihedral angle between the mean planes of the quinoline ring system and the phenyl ring is 48.1 (5)°. The mean plane of the carboxyl-ate group is twisted from the mean planes of the latter by 19.8 (8) and 64.9 (5)°, respectively. The crystal packing features weak C-H⋯O inter-actions, which form chains along [010].

The paradox of an insulating contact between a chemisorbed molecule and a wide band gap semiconductor surface.

Controlling the intrinsic optical and electronic properties of a single molecule adsorbed on a surface requires electronic decoupling of some molecular orbitals from the surface states. Scanning tunneling microscopy experiments and density functional theory calculations are used to study a perylene molecule derivative (DHH-PTCDI), adsorbed on the clean 3 × 3 reconstructed wide band gap silicon carbide surface (SiC(0001)-3 × 3). We find that the LUMO of the adsorbed molecule is invisible in I(V) spectra due to the absence of any surface or bulk states and that the HOMO has a very low saturation current in I(z) spectra. These results present a paradox that the molecular orbitals are electronically isolated from the surface of the wide band gap semiconductor even though strong chemical bonds are formed.

Successful demonstration of an efficient I(-)/(SeCN)2 redox mediator for dye-sensitized solar cells.

A new I(-)/(SeCN)(2) redox mediator has favorable properties for dye-sensitized solar cells (DSCs) such as less visible light absorption, higher ionic conductivity, and downward shift of redox potential than I(-)/I(3)(-). It was then applied for DSCs towards increasing energy conversion efficiency, giving a new potential for improving performance.

Perception of oral health related quality of life (OHQoL-UK) among periodontal risk patients before and after periodontal therapy.

OBJECTIVE: To assess the oral health related quality of life among periodontal risk patients before and after periodontal therapy. METHOD: The study population consisted of 183 patients reporting to the outpatient department of periodontics, who were grouped into low, moderate and high risk patients based on the periodontal risk assessment model. The patients were asked to complete the OHQoL-UK 16 questionnaire before and after periodontal therapy. The change in the patient perception of quality of life before and after treatment was assessed. RESULTS: Periodontitis had a considerable negative impact on the quality of life of patients in the high risk group in comparison to low and moderate risk groups (p < 0.001). Treatment brought about an improvement in the OHQoL scores in the moderate and high risk group. CONCLUSION: This study shows that risk of periodontal disease is significantly related to oral health related quality of life and periodontal therapy improves the quality of life of patients.

Is there need for a transformational change to overcome the current problems with postgraduate medical education in India?

In spite of the existence of a dual system of postgraduation, one under the Medical Council of India (MCI) and the other on a parallel track under the National Board of Examinations, postgraduate medical education in India is beset with several problems. For example, the curriculum has not been revised comprehensively for several decades. The diploma course under the MCI has become unpopular and is largely a temporary refuge for those who do not get admission to degree courses. The level of skills of the outgoing graduate is falling and the increase in the number of seats is taking place in a haphazard manner, without reference to the needs. In spite of increase in seats, there is a shortage of specialists at the secondary and tertiary care levels, especially in medical colleges, to share teaching responsibilities. Further, the distribution of specialists is skewed, with some states having far more than others. To remedy these ills and fulfil the requirements of the country over the next two decades, a working group appointed by the erstwhile governors of the MCI was asked to suggest suitable modifications to the existing postgraduate system. After an extensive review of the lacunae in the present system, the needs at various levels and the pattern of postgraduate education in other countries, it was felt that a competency-based model of a 2-year postgraduate course across all specialties, the use of offsite facilities for training and a criterion-based evaluation system entailing continuous monitoring would go a long way to correct some of the deficiencies of the existing system. The details of the proposal and its merits are outlined for wider discussion and to serve as a feedback to the regulatory agencies engaged in the task of improving the medical education system in India. We feel that the adoption of the proposed system would go a long way in improving career options, increasing the availability of teachers and dissemination of specialists to the secondary and primary levels, and improving the quality of outgoing postgraduates.