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Cyprinid herpesvirus 3 (Cy HV-3) is an alloherpesvirus, and it is the aetiological agent of koi herpesvirus disease. Although the complex morphogenic stages of the replication cycle of Cy HV-3 were shown to resemble that of other members of the Herpesvirales, detailed analysis of the sequence and timing of these events was not definitively determined. This study describes these features through a time course using cyprinid cell cultures ( KF-1 and CCB) infected with Cy HV-3 ( KHV isolate, H361) and analysed by transmission electron microscopy. Rapid viral entry was noted, with high levels of intracellular virus within 1-4 h post-infection (hpi). Intranuclear capsid assembly, paracrystalline array formation and primary envelopment of capsids occurred within 4 hpi. Between 1 and 3 days post-infection (dpi), intracytoplasmic secondary envelopment occurred, as well as budding of infectious virions at the plasma membrane. At 5-7 dpi, the cytoplasm contained cytopathic vacuoles, enveloped virions within vesicles, and abundant non-enveloped capsids; also there was frequent nuclear deformation. Several morphological features are suggestive of inefficient viral assembly, with production of non-infectious particles, particularly in KF-1 cells. The timing of this alloherpesvirus morphogenesis is similar to other members of the Herpesvirales, but there may be possible implications of using different cell lines for Cy HV-3 propagation. [ABSTRACT FROM AUTHOR]

An abstract of the paper "Heterogeneous LPS of Porphyromonas gingivalis differentially modulate the innate immune response of human gingiva," by T. D. K. Herath and colleagues is presented.

Objective: Healthy lifestyle centres (HLCs), a state service that screens for major non-communicable disease (NCD) risk factors and promotes lifestyle modifications in Sri Lanka, report underutilisation. The study aimed to assess the effectiveness of a participatory intervention to empower communities in improving HLC utilisation., Design: A quasi-experimental study based on the principles of community-based participatory research SETTING: Six rural communities each as the intervention (IG) (Gampaha district) and comparison (CG) groups (Kalutara district) from the capital province of Sri Lanka., Participants: Study population was healthy individuals aged 35-65 years, the target group of HLCs in Sri Lanka. A random sample of 498 individuals was selected from each group for evaluation., Interventions: Community support groups (CSGs) were established and empowered using health promotion approach from August 2019 to February 2020. Group discussions and participatory mapping were conducted to identify determinants of underutilisation of HLCs, design activities to address prioritised determinants and develop indicators to monitor the progress of CSGs., Primary and Secondary Outcome Measures: The primary outcome was improvement of HLC utilisation and the secondary outcome was initiation of lifestyle modifications., Results: Significant improvements were seen in the IG, compared with the CG in the seven determinants that contribute to HLC utilisation. The largest differences were seen in reducing negative perceptions of susceptibility for NCDs (pre=64.7%; post=33.3%; p<0.001) and usefulness of screening (pre=66.6%; post=17.3%; p<0.001). The HLC utilisation in IG increased by 29.5% (pre=5.85%; 95% CI 3.74 to 7.95, post=35.3%; 95% CI 30.9 to 39.8, p<0.001), while the utilisation of the CG showed no difference. Furthermore, there was an improvement in the proportion of users who initiated lifestyle modification (pre=64.3%; post=89.9%; p=0.039) in IG, which was not observed in CG., Conclusion: HLC utilisation and initiation of lifestyle modification can be improved by a community-based health promotion intervention through empowering CSGs., Trial Registration Number: SLCTR/2019/028., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Competing Interests: Competing interests: None declared.

The high prevalence of psychological problems observed among healthcare workers (HCWs) during the COVID-19 pandemic called for interventions to safeguard their mental health. We assessed the effectiveness of a 6-week online mindfulness-based intervention in improving well-being and reducing stress among HCWs in Sri Lanka. Eighty HCWs were recruited and randomised into two groups: waitlist-control (WLC) and intervention groups. In the intervention, 1-hour online sessions were conducted at weekly intervals and participants were encouraged to do daily home practice. Stress and well-being were measured pre- and post-intervention using the Perceived Stress Scale and WHO-5 Well-being Index, respectively. One-way analysis of covariance was used to evaluate the effectiveness, in both intention-to-treat (ITT) and complete-case (CC) analyses. A significantly greater improvement in well-being occurred in the intervention arm compared to WLC on both ITT (p = .002) and CC analyses (p < .001), with medium-to-large effect sizes (partial η 2  = .117-.278). However, the reduction in stress following the intervention was not significant compared to the WLC group on both ITT (p = .636) and CC analyses (p = .262). In the intervention arm, the median number of sessions attended by participants was 3. Low adherence to the intervention may have contributed to the apparent non-significant effect on stress., (© 2024 International Union of Psychological Science.)

Introduction. Leptospirosis is a zoonotic disease that is prevalent worldwide. Leptospiral 3-hydroxyacyl-CoA dehydrogenase (3-HADH) is excreted in the urine of infected individuals. However, the potential use of 3-HADH as a biomarker for the diagnosis of leptospirosis using enzyme-linked immunosorbent assay (ELISA) has not been investigated. A technique that identifies Leptospira in a patient in urine sample will be valuable in regular diagnostics and epidemic scenarios, as opposed to existing serological approaches. This study aimed to develop and evaluate an ELISA that can detect 3-HADH in the urine of patients with confirmed acute leptospirosis and to assess its potential as a screening test for leptospirosis. Methods. Laboratory confirmation of acute leptospirosis was done by flaB -nested polymerase chain reaction (PCR) of plasma samples from suspected patients. ELISA-based determination of the presence of 3-HADH in the urine of PCR-positive patients versus PCR-negative patients matched for fever date was performed by coating ELISA plates with urine supernatants and using rabbit anti-3-HADH as the primary antibody. Receiver operating characteristic curve analysis was used to determine the cutoff values for the ELISA. The diagnostic measures between the PCR-positive and PCR-negative patients were compared using the Mann-Whitney U test. Results. In total, 158 febrile patients were assessed, of whom 121 (76.6 %) were male. Of the 15 flaB- nested PCR-positive patients, 12 were in the acute phase of the febrile illness. The best cutoff was an average optical density (OD av ) value of 0.2200 for febrile patients. Sensitivity and specificity were 83.33% [95 % confidence interval (CI), 51.59-97.91 %) and 83.33 % (95 % CI, 76.05-89.13 %), respectively. The OD av values for PCR-positive patients in the acute phase of the disease (≤7 days of fever) were significantly higher than those for PCR-negative patients ( P <0.001, U =114.0, z =-4.946). Conclusion. Detection of 3-HADH in urine by ELISA appears to be promising for the screening of acute leptospirosis in suspected patients., Competing Interests: The University of the Ryukyus currently holds a Japanese patent for the use of a 3-HADH antibody (patent no. 6979200) with C. Toma as the inventor., (Copyright © 2024 The Authors.)

Objective: Accurate estimation of serum thyrotropin (TSH) is crucial in the diagnosis of subclinical hypothyroidism (SCH) in pregnancy. We aimed to investigate whether there are significant differences between fasting and nonfasting thyroid function tests (TFTs) among pregnant mothers., Methods: We studied 100 pregnant mothers with previously unknown thyroid dysfunction. An equal number of participants were included in each trimester. All pregnant mothers underwent fasting and 2-hour postprandial TFTs (TSH, free T4)., Results: Postprandial TSH (mean 1.01 mIU/L, SD 0.80) was significantly lower than the fasting TSH (mean 1.47 mIU/L, SD 1.18) in pregnancy (P < .01). Postprandial free T4 (mean 10.30 pmol/L, SD 2.01) was also lowered compared with fasting free T4 (mean 10.70 pmol/L, SD 1.99) in pregnancy (P < .01). The prevalence of SCH in pregnancy estimated using fasting TSH was 9.4% (SD 3%). In contrast, the prevalence was only 3.5% (SD 2%) when postprandial TSH was used., Conclusion: Compared with the fasting state, postprandial TSH demonstrates a statistically significant reduction that greatly influences the diagnosis of SCH in pregnant mothers. Therefore, we conclude that the timing of sampling for TFTs should be standardized, especially in the pregnant population., (© The Author(s) 2023. Published by Oxford University Press on behalf of American Society for Clinical Pathology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Healthy Lifestyle Centres (HLCs) are state-owned, free-of-charge facilities that screen for major noncommunicable disease risks and promote healthy lifestyles among adults older than 35 years in Sri Lanka. The key challenge to their effectiveness is their underutilisation. This study aimed to describe the underutilisation and determine the factors associated, as a precedent of a bigger project that designed and implemented an intervention for its improvement., Methods: Data derived from a community-based cross-sectional study conducted among 1727 adults (aged 35 to 65 years) recruited using a multi-stage cluster sampling method from two districts (Gampaha and Kalutara) in Sri Lanka. A prior qualitative study was used to identify potential factors to develop the questionnaire which is published separately. Data were obtained using an interviewer-administered questionnaire and analysed using inferential statistics., Results: Forty-two percent (n = 726, 95% CI: 39.7-44.4) had a satisfactory level of awareness on HLCs even though utilisation was only 11.3% (n = 195, 95% CI: 9.80-12.8). Utilisation was significantly associated with 14 factors. The five factors with the highest Odds Ratios (OR) were perceiving screening as useful (OR = 10.2, 95% CI: 4.04-23.4), perceiving as susceptible to NCDs (OR = 6.78, 95% CI: 2.79-16.42) and the presence of peer support for screening and a healthy lifestyle (OR = 3.12, 95% CI: 1.54-6.34), belonging to the second (OR = 3.69, 95% CI: 1.53-8.89) and third lowest (OR = 2.84, 95% CI: 1.02-7.94) household income categories and a higher level of knowledge on HLCs (OR = 1.31, 95% CI: 1.24-1.38). When considering non-utilisation, being a male (OR = 0.18, 95% CI: 0.05-0.52), belonging to an extended family (OR = 0.43, 95% CI: 0.21-0.88), residing within 1-2 km (OR = 0.29, 95% CI: 0.14-0.63) or more than 3 km of the HLC (OR = 0.14, 95% CI: 0.04-0.53), having a higher self-assessed health score (OR = 0.97, 95% CI: 0.95-0.99) and low perceived accessibility to HLCs (OR = 0.12, 95% CI: 0.04-0.36) were significantly associated., Conclusion: In conclusion, underutilisation of HLCs is a result of multiple factors operating at different levels. Therefore, interventions aiming to improve HLC utilisation should be complex and multifaceted designs based on these factors rather than merely improving knowledge., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Herath et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Cryopreserved ram sperm is highly sensitive to oxidative stress by reactive oxygen species (ROS) which impair sperm function and integrity. Antioxidants such as cysteine can mitigate the effect of ROS, although the optimal concentration or timing of supplementation is unknown. This study aimed to determine the effect of concentration and timing of cysteine supplementation on the integrity and function of cryopreserved ram spermatozoa. Nine ejaculates were collected from three Texel rams then cryopreserved and supplemented with cysteine (0, 0.5, or 1.0 mg/mL) added pre-freeze (PF), post-thaw (PT) or pre-freeze and post-thaw (PF + PT) generating seven treatments: 1) control 0 mg/mL, 2) PF 0.5 mg/mL, 3) PF 1 mg/mL, 4) PT 0.5 mg/mL, 5), PT 1.0 mg/mL, 6) PF + PT 0.5 mg/mL and 7) PF + PT 1.0 mg/mL. Sperm motility, viability, acrosome integrity, ROS production and penetrability through artificial cervical mucus were assessed post-thaw. Cysteine supplementation reduced ROS production which thereby improved spermatozoa motility, viability, acrosome integrity and penetrability (p < 0.001) Sperm integrity for all parameters was greatest in spermatozoa treated PF + PT with 1.0 mg/mL cysteine, although treatment pre-freeze or post-thaw also improved integrity beyond the control. This study has identified that 1.0 mg/mL cysteine is most beneficial and has highlighted the importance of preventing oxidative stress in spermatozoa post-thaw. These finding can help to mitigate the detrimental effect of cryopreservation on spermatozoa and aid the development of cryopreservation protocols in sheep., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Competing Interests: Competing interests: None declared.

Purpose: COViK, a prospective hospital-based multicenter case-control study in Germany, aims to assess the effectiveness of COVID-19 vaccines against severe disease. Here, we report vaccine effectiveness (VE) against COVID-19-caused hospitalization and intensive care treatment during the Omicron wave., Methods: We analyzed data from 276 cases with COVID-19 and 494 control patients recruited in 13 hospitals from 1 December 2021 to 5 September 2022. We calculated crude and confounder-adjusted VE estimates., Results: 21% of cases (57/276) were not vaccinated, compared to 5% of controls (26/494; p < 0.001). Confounder-adjusted VE against COVID-19-caused hospitalization was 55.4% (95% CI: 12-78%), 81.5% (95% CI: 68-90%) and 95.6% (95%CI: 88-99%) after two, three and four vaccine doses, respectively. VE against hospitalization due to COVID-19 remained stable up to one year after three vaccine doses., Conclusion: Three vaccine doses remained highly effective in preventing severe disease and this protection was sustained; a fourth dose further increased protection., (© 2023. The Author(s).)

Objective: Healthy lifestyle centres (HLCs) in Sri Lanka provide screening and lifestyle modification services targeting major non-communicable diseases (NCDs). Even though the service is highly accessible and affordable, HLCs are underused by its target population (adults >35 years). We aimed to explore the factors that influence the decision-making process of utilisation of HLCs in Sri Lanka., Setting: Two districts (Gampaha and Kalutara) from the highest populous province (Western) located adjacent to the capital district of Sri Lanka., Participants: Nine service providers, 37 HLC clients and 52 community participants were selected using judgemental, convenient and purposive sampling methods. Theoretical sampling method was used to decide the sample size for each category., Method: A qualitative study design based on constructivist grounded theory was used. Data collected using in-depth interviews and focus group discussions during January to July 2019 and were analysed using the constant comparison method., Results: The decision-making process of utilisation of HLCs was found to be a chain of outcomes with three main steps, such as: intention, readiness and utilisation. Awareness of HLCs, positive attitudes on health, intrinsic or extrinsic motivators, positive attitudes on NCDs and screening were internal factors with a positive influence on intention. Readiness was positively influenced by positive characteristics of the HLCs. It was negatively influenced by negative attitudes on staff and services of HLCs and negative past experiences related to services in state healthcare institutions and HLCs, service provider-related barriers and employment-related barriers. Family-related factors, social support and norms influenced both intention and readiness, either positively or negatively., Conclusion: The decision-making process of utilisation of HLCs links with factors originating from internal, family, service provider and societal levels. Thus, a multifactorial approach that addresses all these levels is needed to improve the utilisation of HLCs in Sri Lanka., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background and Objectives: Tinnitus is a common and disabling condition that largely remains undertreated in Sri Lanka. Currently, standardized tools that assess and monitor the treatment of tinnitus or the distress it causes are unavailable in either of the two main vernacular languages prevalent in Sri Lanka. The Tinnitus Handicap Inventory (THI) is used internationally to measure tinnitus-induced distress and to monitor treatment efficacy. In this study, we validated the Sinhala version of the THI (THI-Sin). Subjects and., Methods: The THI was translated into Sinhala and back translated into English and finalized by independent translators. The THI-Sin questionnaire and the 12-item General Health Questionnaire (GHQ-12) and Visual Analog Scale of tinnitus annoyance (VAS) were administered to 122 adults who visited the otolaryngology clinic of Colombo North Teaching Hospital, Ragama, Sri Lanka., Results: THI-Sin scores showed satisfactory internal consistency (Cronbach's α=0.902) and were significantly correlated with the GHQ-12 and VAS scores. Factor analysis of the THI-Sin confirmed a three-factorial structure, which did not correspond to the original THI subscales., Conclusions: We observed significant reliability and validity of the THI-Sin tool for evaluation of tinnitus-induced handicaps among the Sinhalese-speaking population of Sri Lanka.

Objectives: The COVID-19 vaccine is currently being administered worldwide to address the ongoing pandemic. Although these vaccines have proven effective in preventing severe disease, the level of immunity required to prevent respiratory mucosal infection remains less well understood. Therefore, it is desirable to develop a noninvasive screening strategy such as oral fluid to monitor secreted antibodies longitudinally as potential surrogates of mucosal immunity., Methods: We evaluated the anti-spike protein antibodies in gingival crevicular fluid (GCF) and saliva and compared them to immune responses in the blood of 50 healthy health care workers following 2 doses of intramuscular Pfizer/BioNTech-BNT162b2 vaccine., Results: The antibodies to SARS-CoV-2 spike and subdomain proteins (RBD, S1, S2, and NTD) were significantly higher in serum than oral fluids but showed a greater detection rate and higher median titres in GCF than saliva. For all tested SARS-CoV-2 antigens, IgG in GCF (as opposed to saliva) showed a more significant and stronger correlation with IgG in serum. Serum-neutralising antibodies (Nab) titres also displayed a significant and stronger correlation with anti-spike protein and their subdomains in GCF than saliva. Interestingly, the time post-second dose of vaccine and sex had a similar influence on IgG in serum and GCF. However, interferon (IFN)-γ-producing T-cell responses showed no association with SARS-Cov-2 IgG antibodies in serum, GCF, or saliva and neutralisation antibodies in serum. The correlation matrix of all measured parameters grouped serum and GCF IgG parameters separately from salivary IgG parameters indicating that GCF better represents the humoural response in serum than saliva., Conclusions: Within limitations, we propose that GCF could be a less invasive alternative to serum and more appropriate than saliva to detect antibody responses by current COVID-19 vaccines if the GCF collection procedure could be standardised. Further research is needed to investigate the suitability of GCF for community immune surveillance for vaccines., Competing Interests: Conflict of interest None disclosed., (Copyright © 2022. Published by Elsevier Inc.)

A 42-year-old man developed bilateral Tapia's syndrome (recurrent laryngeal and hypoglossal nerves paralysis) following prolonged ventilation for COVID-19 pneumonia. Examination showed global tongue atrophy and bilateral asymmetric vocal cord palsy. He improved rapidly without specific treatment, suggesting that neuropraxia was the likely mechanism of injury. Tapia's syndrome has been reported disproportionately more often in association with COVID-19, possibly from injury to hypoglossal and vagal nerves during ventilation in the prone position., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)