Initial Infection Research Articles

Monophasic Variant of Salmonella Typhimurium Infection Affects the Serum Metabolome in Swine.

Salmonella is the most relevant foodborne zoonotic agent found in swine, and its presence in French herds is significant. Its carriage is asymptomatic, which makes it difficult to detect during rearing, thus increasing the risk of its presence on pork meat. Studies have shown that enteric infection in animals could be associated with changes in the serum metabolome composition, through the immune response or changes in the digestive microbiota composition. We hypothesized that these changes in the serum metabolome composition could be used as markers for the detection of asymptomatic animals infected by Salmonella. Using untargeted analysis by liquid chromatography coupled with mass spectrometry, we showed that significant differences in the composition of the serum metabolome could be detected between infected or noninfected animals both 1 and 21 days after experimental infection. This serum metabolome composition significantly changed during the 21 days postinfection in the infected animal groups, suggesting an evolution of the impact of infection with time. Despite this evolution, differences in the serum metabolome composition persisted between infected and noninfected animals 21 days after the initial infection. We also showed a possible difference between high-shedding and low-shedding animals 21 days postinfection. Finally, some of the variations in the metabolome were found to be significantly associated with variations of specific members of the fecal microbiota. Thus, excreting and asymptomatic animals, but also high-shedding animals, could be identified on the basis of their serum metabolome composition.

Sexually Transmitted Infections and the Risk of Reinfection Within 12 Months: A Population-Based Cohort.

Chlamydia, gonorrhea, and syphilis are common sexually transmitted infections that disproportionately affect specific groups in New Zealand (NZ). Predictors of reinfection are not well studied in NZ but could inform public health strategies to decrease sexually transmitted infection (STI) incidence. New Zealand-wide chlamydia, gonorrhea, and syphilis cases during 2019 were identified using nationally collected data. Cases were followed-up to identify reinfection with the same STI within 12 months of initial infections. Logistic regression models were used to identify predictors for each STI reinfection. Determinants identified for increased odds of chlamydia reinfection were age groups 16-19 and 20-24 years, females, Māori and Pacific peoples, cases in the Northern region, and cases with at least one test before the initial infection. Age 40 years and older was associated with lower odds of gonorrhea reinfection, as was being of Asian ethnicity, living in Midland or Southern regions, and reporting heterosexual behavior. Region was the only statistically significant predictor for syphilis reinfection, with higher odds of reinfection for people living in the Central region. Our findings reflect disproportionate STI rates for some groups in NZ, with younger age groups, Māori and Pacific peoples, men who have sex with men, and people living in the Northern region experiencing higher odds of reinfection. Groups identified with higher odds for reinfection require increased access to culturally responsive health services to treat, understand, and prevent possible reinfection. Changes to current public health strategies could include culturally specific behavioral counseling, and improvements to and adherence to effective contract tracing.

Improving the Predictive Value of Preclinical Mouse Models of Pseudomonas aeruginosa Respiratory Infection to Evaluate Antibiotic Efficacy.

Disease-specific animal models and treatment regimens are essential to optimize anti-Pseudomonas drug testing. Mouse models of acute and chronic P. aeruginosa infections provide valuable resources that mimic the initial and progressive bronchopulmonary infection typically observed in a wide range of patients - in hospital settings, with cystic fibrosis or chronic obstructive pulmonary disease. In this chapter, we will explain how mice can be treated using different administration routes in disease-specific models of P. aeruginosa respiratory infection. We will also describe methods used to evaluate multiple endpoints, including profiling of bacterial and host responses. The application of these procedures in disease-specific models is essential to optimize the efficacy of anti-P. aeruginosa treatments and provide an enhanced link between preclinical testing and clinical trials.

Cytometry profiling of ex vivo recall responses to Coxiella burnetii in previously naturally exposed individuals reveals long-term changes in both adaptive and innate immune cellular compartments.

Q fever, caused by the intracellular bacterium Coxiella burnetii, is considered an occupational and biodefense hazard and can result in debilitating long-term complications. While natural infection and vaccination induce humoral and cellular immune responses, the exact nature of cellular immune responses to C. burnetii is incompletely understood. The current study seeks to investigate more deeply the nature of long-term cellular recall responses in naturally exposed individuals by both cytokine release assessment and cytometry profiling. Individuals exposed during the 2007-2010 Dutch Q fever outbreak were grouped in 2015, based on a C. burnetii-specific IFNγ release assay (IGRA), serological status, and self-reported clinical symptoms during initial infection, into asymptomatic IGRA-negative/seronegative controls, and three IGRA-positive groups (seronegative/asymptomatic; seropositive/asymptomatic and seropositive/symptomatic). Recall responses following in vitro re-stimulation with heat-inactivated C. burnetii in whole blood, were assessed in 2016/2017 by cytokine release assays (n=55) and flow cytometry (n=36), and in blood mononuclear cells by mass cytometry (n=36). Cytokine release analysis showed significantly elevated IL-2 responses in all seropositive individuals and elevated IL-1β responses in those recovered from symptomatic infection. Comparative flow cytometry analysis revealed significantly increased IFNγ, TNFα and IL-2 recall responses by CD4 T cells and higher IL-6 production by monocytes from symptomatic, IGRA-positive/seropositive individuals compared to controls. Mass cytometry profiling and unsupervised clustering analysis confirmed recall responses in seropositive individuals by two activated CD4 T cell subsets, one characterized by a strong Th1 cytokine profile (IFNγ+IL-2+TNFα+), and identified C. burnetii-specific activation of CD8 T cells in all IGRA-positive groups. Remarkably, increased C. burnetii-specific responses in IGRA-positive individuals were also observed in three innate cell subpopulations: one characterized by an IFNγ+IL-2+TNFα+ Th1 cytokine profile and lack of canonical marker expression, and two IL-1β-, IL-6- and IL-8-producing CD14+ monocyte subsets that could be the drivers of elevated secretion of innate cytokines in pre-exposed individuals. These data highlight that there are long-term increased responses to C. burnetii in both adaptive and innate cellular compartments, the latter being indicative of trained immunity. These findings warrant future studies into the protective role of these innate responses and may inform future Q fever vaccine design.

Immunomodulatory effect of Ti-Cu alloy by surface nanostructure synergistic with Cu2+ release

The inflammatory response induced by implant/macrophage interaction has been considered to be one of the vital factors in determining the success of implantation. In this study, TiCuNxOy coating with an immunomodulatory strategy was proposed for the first time, using nanostructured TiCuNxOy coating synthesized on Ti-Cu alloy by oxygen and nitrogen plasma-based surface modification. It was found that TiCuNxOy coating inhibited macrophage proliferation but stimulated macrophage preferential activation and presented an elongated morphology due to the surface nanostructure. The most encouraging discovery was that TiCuNxOy coating promoted the initial pro-inflammatory response of macrophages and then accelerated the M1-to-M2 transition of macrophages via a synergistic effect of fast-to-slow Cu2+ release and surface nanostructure, which was considered to contribute to initial infection elimination and tissue healing. As expected, TiCuNxOy coating released desirable Cu2+ and generated a favorable immune response that facilitated HUVEC recruitment to the coating, and accelerated proliferation, VEGF secretion and NO production of HUVECs. On the other hand, it is satisfying that TiCuNxOy coating maintained perfect long-term antibacterial activity (≥99.9%), mainly relying on Cu2O/CuO contact sterilization. These results indicated that TiCuNxOy coating might offer novel insights into the creation of a surface with immunomodulatory effects and long-term bactericidal potential for cardiovascular applications.

Comprehensive Risk Assessment of Infection Induced by SARS-CoV-2.

The pandemic COVID-19 (coronavirus disease 2019) is caused by the severe acute respiratory syndrome corona virus 2 (SARS-CoV-2), which devastated the global economy and healthcare system. The infection caused an unforeseen rise in COVID-19 patients and increased the mortality rate globally. This study gives an overall idea about host-pathogen interaction, immune responses to COVID-19, recovery status of infection, targeted organs and complications associated, and comparison of post-infection immunity in convalescent subjects and non-infected individuals. The emergence of the variants and episodes of COVID-19 infections made the situation worsen. The timely introduction of vaccines and precautionary measures helped control the infection's severity. Later, the population that recovered from COVID-19 grew significantly. However, understanding the impact of healthcare issues resulting after infection is paramount for improving an individual's health status. It is now recognised that COVID-19 infection affects multiple organs and exhibits a broad range of clinical manifestations. So, post COVID-19 infection creates a high risk in individuals with already prevailing health complications. The identification of post-COVID-19-related health issues and their appropriate management is of greater importance to improving patient's quality of life. The persistence, sequelae and other medical complications that normally last from weeks to months after the recovery of the initial infection are involved with COVID-19. A multi-disciplinary approach is necessary for the development of preventive measures, techniques for rehabilitation and strategies for clinical management when it comes to long-term care.

Wolbachia invasion to wild mosquito population in stochastic environment

Releasing Wolbachia-infected mosquitoes to invade the wild mosquito population is a method of mosquito control. In this paper, a stochastic mosquito population model with Wolbachia invasion perturbed by environmental fluctuation is studied. Firstly, the well-posedness, positivity, and Markov-Feller property of the solution for this model are proved. Then a group of sharp threshold-type conditions is provided to characterize the long-term behavior of the model, which pinpoints the almost necessary and sufficient conditions for the persistence and extinction of Wolbachia-infected and uninfected mosquito populations. Our results indicate that even for a low initial Wolbachia infection frequency, a successful Wolbachia invasion into the wild mosquito population can be driven by stochastic environmental fluctuations. Finally, some numerical experiments are carried out to support our theoretical results.

Low growth hormone secretion associated with post-acute sequelae SARS-CoV-2 infection (PASC) neurologic symptoms: A case-control pilot study

ObjectiveTo determine if patients that develop lingering neurologic symptoms of fatigue and “brain fog” after initial recovery from coronavirus disease 2019 (COVID-19) have persistent low growth hormone (GH) secretion as seen in other conditions with similar symptom etiology. DesignIn this case-control observational pilot study, patients reporting lingering neurologic post-acute sequelae of SARS-CoV-2 (PASC, n = 10) symptoms at least 6 months after initial infection were compared to patients that recovered from COVID-19 without lingering symptoms (non-PASC, n = 13). We compared basic blood chemistry and select metabolites, lipids, hormones, inflammatory markers, and vitamins between groups. PASC and non-PASC subjects were tested for neurocognition and GH secretion, and given questionnaires to assess symptom severity. PASC subjects were also tested for glucose tolerance and adrenal function. ResultsPASC subjects reported significantly worse fatigue, sleep quality, depression, quality of life, and gastrointestinal discomfort compared to non-PASC. Although PASC subjects self-reported poor mental resilience, cognitive testing did not reveal significant differences between groups. Neurologic PASC symptoms were not linked to inflammatory markers or adrenal insufficiency, but were associated with reduced growth hormone secretion. ConclusionsNeurologic PASC symptoms are associated with gastrointestinal discomfort and persistent disruption of GH secretion following recovery from acute COVID-19. (www.clinicaltrials.gov; NCT04860869)

Over-expression of IL-33 enhances rabies virus early antigen presentations and cellular immune responses in mice

Human inactivated rabies virus (RABV) vaccines have been widely used worldwide over 30 years. The mechanisms of humoral immunity elicited by previously reported rabies candidate vaccines have been fully investigated, but little is known about the cellular immunity profiles. Herein, the recombinant RABV rLBNSE-IL-33 overexpressing the mouse interleukin-33 (IL-33) proliferated well in Neuro-2a cells and had no effects with the parent virus on growth kinetic in vitro and viral pathogenicity in mice. The rLBNSE-IL-33 experienced more antigen presentations by MHC-II on DCs and activated more CD4+ T cells which helped recruit more CD19+CD40+ B cells in blood and promote rapid and robust IgG1 antibodies responses at initial infection stage compared with the parent rLBNSE strain. Simultaneously, the rLBNSE-IL-33 were also presented by MHC-I to CD8+ T cells which contributed to produce high levels of IgG2a. The rLBNSE-IL-33 elicited significantly high levels of RABV-specific IFN-γ secreting memory CD4+ T cells, more RABV-specific IL-4 and IFN-γ secreting memory CD8+ T cells in spleens at early infection stage in mice. Altogether, overexpression of IL-33 in rLBNSE-IL-33 enhanced early antigen presentation, markedly promote CD4+, memory CD4+ and CD8+ T cells-mediated responses and provided a 100 % protection from lethal RABV challenge in mice. These findings provided an alternative novel therapy and vaccine strategy in future.

First Report of Leaf Spot Caused by Phyllosticta styracicola on Nageia nagi in China.

Nageia nagi (Thunb.) Kuntze belongs to the family Podocarpaceae with shiny green branches and leaves, which is widely distributed in East Asia and the Southern Hemisphere. The leaves, roots and fruits of N. nagi have been used as herbal medicine to treat rheumatism, arthritis and venereal diseases (Abdillahi et al. 2011). In September 2022, leaf spot symptoms were found on approximately 30% of the leaves of N. nagi trees in a community located at the Economic and Technological Development Zone, Nanchang City, Jiangxi Province, China. Following the initial infection, the leaf lesions extended outwards from the top in a circular pattern, appearing as a dark brick color, and later changed to yellow and became dry, with a darker brown margin surrounding them. Ten symptomatic leaves were randomly selected, and a small piece of leaf tissue (5mm ×5mm) located between the health and infected tissues was cut and surface-desinfected with 70% ethanol for 30 s and 1% sodium hypochlorite (NaClO) for 30 s sequentially. After rinsing three times in sterile distilled water, all the small pieces of leaves were placed on potato dextrose agar (PDA) plates, followed by incubation at 28℃ for 3 days. Ten isolates, cultured on each PDA plate, appeared olive green with a granular surface, and an uneven white edge, and finally turned greenish black. The conidia were hyaline, with ellipsoidal to subglobose shapes and spore sizes of 5.5-8.3 × 7.2-12.0 μm (width × length) (=7.2±0.71 × 9.9±1.3 μm, n=40). These morphological characteristics are consistent with those of Phyllosticta species. To confirm the species, three representative isolates, JFRL 03-768, JFRL 03-769 and JFRL 03-770 were selected for further identification. The internal transcribed spacer (ITS) region, actin (ACT), translation elongation factor 1-alpha (TEF1-a), and glyceradehyde-3-phosphate dehydrogenase (GPD) genes of the three isolates were amplified and sequenced with the primers V9G/ITS4 (Carbone and Kohn 1999), ACT-512F/ACT-783R (Carbone and Kohn 1999), EF-728F/EF-2 (O´Donnell et al. 1998) and Gpd1-LM/Gpd2-LM (Myllys et al. 2002; Guerber et al. 2003), respectively. All sequences had been deposited into GenBank (ITS: OQ195332, OQ195333 and OQ195334; ACT: OQ207621, OQ207622 and OQ207623; TEF1-a: OQ207624, OQ207625 and OQ207626; GPD: OQ207627, OQ207628 and OQ207629). A maximum likelihood phylogenetic tree was constructed using the IQtree V1.5.6 (Ngugen et al. 2015) based on the concatenation of multiple sequences (ITS, ACT, TEF1-a and GPD). In the cluster analysis, the representative isolates (JFRL 03-768, JFRL 03-769 and JFRL 03-770) were positioned within a clade comprising of Phyllosticta styracicola. Subsequently, the pathogenicity of P. styracicola was determined by wound inoculation of healthy 2 year-old N. nagi plants, and this experiment was repeated for three times. Briefly, for each isolates, six disinfected leaves were wounded with a sterile scalpel, and then inoculated with 10-μl drop of the conidial suspension (1 × 106 conidia/ml). Another six disinfected leaves were inoculated with 10-μl drop of sterile water as a control group, and all plants were incubated at 28°C with 80% humidity. After 15 days, a similar spot lesion appeared on the leaves of the experimental group. P. styracicola was successfully re-isolated, and then subjected to morphological identification and molecular sequencing (ITS, ACT, TEF1-a and GPD genes). Whilst, the control leaves showed no symptoms. Previous studies have reported that P. styracicola could result in the development of lesions on Styrax grandiflorus leaves in China (Zhang et al. 2013). To our knowledge, this is the first report that P. styracicola can cause leaf spot on N. nagi in China.

Influenza enhances host susceptibility to non-pulmonary invasive Streptococcus pyogenes infections

ABSTRACT Streptococcus pyogenes (group A streptococcus; GAS) causes a variety of invasive diseases (iGAS) such as bacteremia, toxic shock syndrome, and pneumonia, which are associated with high mortality despite the susceptibility of the bacteria to penicillin ex vivo. Epidemiologic studies indicate that respiratory influenza virus infection is associated with an increase in the frequency of iGAS diseases, including those not directly involving the lung. We modified a murine model of influenza A (IAV)-GAS superinfection to determine if viral pneumonia increased the susceptibility of mice subsequently infected with GAS in the peritoneum. The results showed that respiratory IAV infection increased the morbidity (weight loss) of mice infected intraperitoneally (i.p.) with GAS 3, 5, and 10 d after the initial viral infection. Mortality was also significantly increased when mice were infected with GAS 3 and 5 d after pulmonary IAV infection. Increased mortality among mice infected with virus 5 d prior to bacterial infection correlated with increased dissemination of GAS from the peritoneum to the blood, spleen, and lungs. The interval was also associated with a significant increase in the pro-inflammatory cytokines IFN-γ, IL-12, TNF-α, MCP-1 and IL-27 in sera. We conclude, using a murine model, that respiratory influenza virus infection increases the likelihood and severity of systemic iGAS disease, even when GAS infection does not originate in the respiratory tract.

Secondary infections in critically ill patients with COVID-19 receiving steroid therapy.

Secondary infections can occur during or after the treatment of an initial infection. Glucocorticoids may decrease mortality in patients with severe COVID-19; however, risk of secondary infection is not well described. Our primary objective was to investigate the risk of secondary infection among critically ill patients with COVID-19 treated with glucocorticoids. We examined patients with COVID-19 being treated in the intensive care unit at two academic medical centers from 1 to 7/2020. One hundred-seven patients were included. Of these, 31 received steroids and 76 patients did not. Analysis of the larger cohort was performed followed by a matched pairs analysis of 22 steroid and 22 non-steroid patients. Secondary infection was seen in 14 patients (45.2%) receiving steroids compared to 35(46.1%) not receiving steroids (p = 0.968). Secondary infections were most frequently encountered in the respiratory tract. Escherichia coli and Staphylococcus aureus were the most frequently identified organisms. Mortality was 16.1% in the steroid-treated group compared to 23.7% in the control group (p = 0.388). After performing matched pairs analysis and multivariable logistic regression there was no significant difference between secondary infection or mortality and steroid receipt. Secondary infections were common among critically ill patients with COVID-19, but the incidence of secondary infection was not significantly impacted by steroid treatment.

Periprosthetic Joint Infections of the Knee Lastingly Impact the Bone Homeostasis.

After periprosthetic joint infection (PJI)-dependent revision surgery, a significantly elevated number of patients suffer from prosthesis failure due to aseptic loosening and require additional revision surgery despite clearance of the initial infection. The mechanisms underlying this pathology are not well understood, as it has been assumed that the bone stock recovers after revision surgery. Despite clinical evidence suggesting decreased osteogenic potential in PJI, understanding of the underlying biology remains limited. In this study, we investigated the impact of PJI on bone homeostasis in a two-stage exchange approach at explantation and reimplantation. Sixty-four human tibial and femoral specimens (20 control, 20 PJI septic explantation, and 24 PJI prosthesis reimplantation samples) were analyzed for their bone microstructure, cellular composition, and expression of relevant genetic markers. Samples were analyzed using X-ray microtomography, Alcian blue and tartrate-resistant acid phosphatase staining, and RT-qPCR. In patients with PJI, bone volume (BV/TV; 0.173 ± 0.026; p < 0.001), trabecular thickness (164.262 ± 18.841 μm; p < 0.001), and bone mineral density (0.824 ± 0.017 g/cm2 ; p = 0.049) were reduced; trabecular separation (1833.939 ± 178.501 μm; p = 0.005) was increased. While prevalence of osteoclasts was elevated (N.Oc/BS: 0.663 ± 0.102, p < 0.001), osteoblast cell numbers were lower at explantation (N.Ob/BS: 0.149 ± 0.021; p = 0.047). Mean expression of bone homeostasis markers osteocalcin, osteopontin, Runx2, TSG-6, and FGF-2 was significantly reduced at prosthesis explantation. Despite partial recovery, all analyzed parameters were still significantly impacted at reimplantation. In contrast, mean expression of osteoclastogenesis-stimulating cytokine IL-17a was significantly increased at both explantation and reimplantation. In this study, we found a strong and lasting impact of PJI on the bone homeostasis on a molecular, cellular, and microstructural level. These changes may be responsible for the increased risk of prosthesis failure due to aseptic loosening. Our data suggest there is significant potential in modulating bone homeostasis to improve prosthesis fixation and long-term clinical outcome in affected patients. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

A descriptive study of post-COVID sequelae in mild to severe cases in western India

Abstract Background: COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a multisystem illness that has affected millions worldwide in the form of a pandemic. Numerous studies have shown that individuals infected with SARS-CoV-2 experienced extended periods of health issues, varying from fatigue to cardiovascular and respiratory complications, lasting from 12 weeks to 1 year. The prevalence of these issues ranged between 42% and 55% in different countries, correlating with the severity of the initial infection. Therefore, this study aimed to evaluate the impact of SARS-CoV-2 infection consequences and identify the factors linked to these long-term effects. Materials and Methods: A cross-sectional study was carried out at a tertiary care hospital over 3 months, from Jun 14, 2021 to Sept 30, 2021. This study focused on three groups of patients—those with SARS-CoV-2 infection treated in outpatient departments (OPD) and hospitalized individuals—observed for follow-up at 3, 6, and 9 months postinfection. With verbal consent obtained after informed discussion, information regarding demographic details, pre-existing medical conditions, habits, and vaccination status was gathered via telephonic interviews using a semistructured questionnaire. The collected data underwent analysis utilizing MS Excel and Open EPI info software. Results: In the follow-up studies at 3, 6, and 9 months, there were 1902 patients (with a response rate of 54.06%), 876 patients (with a response rate of 64.75%), and 1464 patients (with a response rate of 58.04%), respectively. The prevalence of post-COVID sequelae across these periods was recorded at 8.20%, 5.59%, and 8.81%. The mortality rate was notably 7.46% within the 3-month follow-up cohort. Diabetes emerged as a prevalent sequela across all three follow-up cohorts, peaking at 2.86% in the 9-month cohort. Conclusion: It is necessary to conduct 1-year post-COVID-19 infection patient follow-ups.

Reproductive strategies of the parasitic flatworm Thaparocleidus vistulensis (Siwak, 1932) (Platyhelminthes, Monogenea) infecting the European catfish Silurus glanis Linnaeus, 1758

The life cycle of Thaparocleidus vistulensis (Siwak, 1932), a host-specific monogenean parasite of European catfish (Silurus glanis Linnaeus, 1758), was investigated by detailed observation of infection dynamics, egg development, hatching rate and in vitro survival rates of the parasite at different life stages at 23 °C. A total of 30 naive fingerlings were infected in three exposure trials by co-habitation with donor fish carrying adult parasites. Two fish were dissected every two days during the 10-day experimental period to explore the development of larvae and juvenile parasites on the host gills. Freshly laid eggs by adult monogeneans were collected and observed daily under a light microscope until hatching. A total of 445 eggs were collected and distributed into wells of 96-well microtiter plates containing filtered fish tank water to determine their hatching rates. A similar method was used to investigate the survival rates of isolated parasites at different developmental stages (larvae, juveniles, and adults). T. vistulensis populations on the European catfish in fish tanks increased markedly within ten days, dependent on the severity of the initial infection levels of the donor fish. The first eggs hatched three to four days after oviposition, and the hatching rate peaked on the fifth day (89.7%). The survival rate for freely swimming oncomiracidia without host was 7.4% after five days, whereas isolated juvenile and adult parasites showed a higher dependence of host contact (survival rates three days post-isolation of 0.9% and 1.6%, respectively). The data allows prediction of parasite-host dynamics and may improve control of gill-disease in cultured European catfish stocks in fish farms.

Focal Adhesion Kinase Binds to the HPV E2 Protein to Regulate Initial Replication after Infection.

Human papillomaviruses are small DNA tumor viruses that infect cutaneous and mucosal epithelia. The viral lifecycle is linked to the differentiation status of the epithelium. During initial viral infection, the genomes replicate at a low copy number but the mechanism(s) the virus uses to control the copy number during this stage is not known. In this study, we demonstrate that the tyrosine kinase focal adhesion kinase (FAK) binds to and phosphorylates the high-risk viral E2 protein, the key regulator of HPV replication. The depletion of FAK with a specific PROTAC had no effect on viral DNA content in keratinocytes that already maintain HPV-16 and HPV-31 episomes. In contrast, the depletion of FAK significantly increased HPV-16 DNA content in keratinocytes infected with HPV-16 quasiviruses. These data imply that FAK prevents the over-replication of the HPV genome after infection through the interaction and phosphorylation of the E2 protein.

Cochleo-Vestibular Disorders in Herpes Zoster Oticus: A Literature Review and a Case of Bilateral Vestibular Hypofunction in Unilateral HZO.

The varicella-zoster virus (VZV), a member of the Herpesviridae family, causes both the initial varicella infection and subsequent zoster episodes. Disorders of the eighth cranial nerve are common in people with herpes zoster oticus (HZO). We performed a review of the literature on different databases including PubMed and SCOPUS, focusing on cochlear and vestibular symptoms; 38 studies were considered in our review. A high percentage of cases of HZO provokes cochlear and vestibular symptoms, hearing loss and vertigo, whose onset is normally preceded by vesicles on the external ear. It is still under debate if the sites of damage are the inferior/superior vestibular nerves and cochlear nerves or a direct localization of the infection in the inner ear. The involvement of other contiguous cranial nerves has also been reported in a few cases. We report the case of a patient with single-side HZO presenting clinical manifestations of cochleo-vestibular damage without neurological and meningeal signs; after 15 days, the patient developed a new episode of vertigo with clinical findings of acute contralateral vestibular loss. To our knowledge, only three other such cases have been published. An autoimmune etiology may be considered to explain these findings.

Experimental infection and co-infection with Chinese strains of Ehrlichia canis and Babesia vogeli in intact and splenectomized dogs: Insights on clinical, hematologic and treatment responses

Animal infection models are crucial for studying various aspects of Ehrlichia canis infections. To understand the pathogenesis of the first Chinese isolate of E. canis and simulate the natural progression of canine ehrlichiosis, we developed a model with 18 Beagle dogs that consisted of E. canis initial infection (days 0–17), treatment with doxycycline or rifampicin (days 18–32), recovery (days 33–66), E. canis reinfection (days 67–91), and Babesia vogeli superinfection (days 92–116). We measured body weight and rectal temperature every other day, drew blood every 4 days for routine hematology and biochemistry tests, and for quantification of E. canis and B. vogeli by quantitative PCRs. In this study, the first isolate of E. canis from China was used to experimentally infect dogs, and the infected dogs exhibited clinical signs of acute severe ehrlichiosis, including high fever, loss of appetite, dehydration, and body weight loss, confirming the similar pathogenicity of E. canis in China as compared to isolates from other regions. Infection with E. canis and B. vogeli led to reduced body weight and fever in dogs. Doxycycline treatment led to absence of E. canis DNA in infected dogs, while rifampicin treatment lowered the blood E. canis copy number up to 1.5 folds. E. canis-free infected dogs after doxycycline treatment were successfully re-infected with E. canis, indicating dogs with antibodies are still at risk of re-infection. Super-infection with B. vogeli resulted in higher fever, more severe anemia, and a reduced number of platelets. Splenectomized dogs showed significantly higher E. canis numbers during recovery and re-infection than intact dogs. The histological changes were observed in brain, lung, kidney, liver and spleen of the infected dogs. The findings in this study provide insights into clinical and hematologic responses, as well as effective treatment options, for dogs infected with the first Chinese isolate of E. canis, and may contribute to our understanding of the diagnosis and prevention of tick-borne diseases in dogs, including canine monocytic ehrlichiosis.

Characterization of integrated Marek's disease virus genomes supports a model of integration by homology-directed recombination and telomere-loop-driven excision.

Marek's disease virus (MDV) is a ubiquitous chicken pathogen that inflicts a large economic burden on the poultry industry, despite worldwide vaccination programs. MDV is only partially controlled by available vaccines, and the virus retains the ability to replicate and spread between vaccinated birds. Following an initial infection, MDV enters a latent state and integrates into host telomeres and this may be a prerequisite for malignant transformation, which is usually fatal. To understand the mechanism that underlies the dynamic relationship between integrated-latent and reactivated MDV, we have characterized integrated MDV (iMDV) genomes and their associated telomeres. This revealed a single orientation among iMDV genomes and the loss of some terminal sequences that is consistent with integration by homology-directed recombination and excision via a telomere-loop-mediated process.

Effect of primaquine dose on the risk of recurrence in patients with uncomplicated Plasmodium vivax: a systematic review and individual patient data meta-analysis

Primaquine is used to eliminate Plasmodium vivax hypnozoites, but its optimal dosing regimen remains unclear. We undertook a systematic review and individual patient data meta-analysis to investigate the efficacy and tolerability of different primaquine dosing regimens to prevent P vivax recurrence. For this systematic review and individual patient data meta-analysis, we searched MEDLINE, Web of Science, Embase, and Cochrane Central for prospective clinical studies of uncomplicated P vivax from endemic countries published between Jan 1, 2000, and June 8, 2023. We included studies if they had active follow-up of at least 28 days, and if they included a treatment group with daily primaquine given over multiple days, where primaquine was commenced within 7 days of schizontocidal treatment and was given alone or coadministered with chloroquine or one of four artemisinin-based combination therapies (ie, artemether-lumefantrine, artesunate-mefloquine, artesunate-amodiaquine, or dihydroartemisinin-piperaquine). We excluded studies if they were on prevention, prophylaxis, or patients with severe malaria, or if data were extracted retrospectively from medical records outside of a planned trial. For the meta-analysis, we contacted the investigators of eligible trials to request individual patient data and we then pooled data that were made available by Aug 23, 2021. We assessed the effects of total dose and duration of primaquine regimens on the rate of first P vivax recurrence between day 7 and day 180 by Cox's proportional hazards regression (efficacy analysis). The effect of primaquine daily dose on gastrointestinal symptoms on days 5-7 was assessed by modified Poisson regression (tolerability analysis). The study was registered with PROSPERO, CRD42019154470. Of 226 identified studies, 23 studies with patient-level data from 6879 patients from 16 countries were included in the efficacy analysis. At day 180, the risk of recurrence was 51·0% (95% CI 48·2-53·9) in 1470 patients treated without primaquine, 19·3% (16·9-21·9) in 2569 patients treated with a low total dose of primaquine (approximately 3·5 mg/kg), and 8·1% (7·0-9·4) in 2811 patients treated with a high total dose of primaquine (approximately 7 mg/kg), regardless of primaquine treatment duration. Compared with treatment without primaquine, the rate of P vivax recurrence was lower after treatment with low-dose primaquine (adjusted hazard ratio 0·21, 95% CI 0·17-0·27; p<0·0001) and high-dose primaquine (0·10, 0·08-0·12; p<0·0001). High-dose primaquine had greater efficacy than low-dose primaquine in regions with high and low relapse periodicity (ie, the time from initial infection to vivax relapse). 16 studies with patient-level data from 5609 patients from ten countries were included in the tolerability analysis. Gastrointestinal symptoms on days 5-7 were reported by 4·0% (95% CI 0·0-8·7) of 893 patients treated without primaquine, 6·2% (0·5-12·0) of 737 patients treated with a low daily dose of primaquine (approximately 0·25 mg/kg per day), 5·9% (1·8-10·1) of 1123 patients treated with an intermediate daily dose (approximately 0·5 mg/kg per day) and 10·9% (5·7-16·1) of 1178 patients treated with a high daily dose (approximately 1 mg/kg per day). 20 of 23 studies included in the efficacy analysis and 15 of 16 in the tolerability analysis had a low or unclear risk of bias. Increasing the total dose of primaquine from 3·5 mg/kg to 7 mg/kg can reduce P vivax recurrences by more than 50% in most endemic regions, with a small associated increase in gastrointestinal symptoms. Australian National Health and Medical Research Council, Bill & Melinda Gates Foundation, and Medicines for Malaria Venture.