Cell proliferation and apoptosis in canine oral papillomatosis.

This study was aimed at the evaluation of cell proliferation, p53 level and apoptotic index by immunohistochemical methods in canine oral papillomatosis. The study material comprised of tumor tissue samples taken from six dogs being admitted to the Pathology Department of Faculty of Veterinary Medicine, Kafkas University, Kars, Türkiye. Choice of immunohistochemical staining was avidin-biotin peroxidase method. Cases of canine oral papillomatosis, determined to have been caused by canine papillomavirus-1, were found to have a rather high cell proliferation index. Furthermore, all cases were immunohisto-chemically demonstrated to carry a mutant p53 gene. Despite the mutation of p53 gene, the shift in the Bax/Bcl-2 ratio of dogs diagnosed with tumor was in favor of the pro-apoptotic Bax gene. The apoptotic mechanism was determined to occur through both the caspase-dependent and caspase-independent pathways. While the lesions occupied the entire oral cavity in some cases, histopathologically, malignant transformation was not detected in any of the six cases.

Identification and molecular characterization of two papillomavirus genotypes in teat papillomatosis cases in hair goats, in Türkiye.

Papillomaviruses can cause benign or malignant proliferations on the host's skin and mucous membranes. Recent genetic studies have identified many new papillomavirus types. In this study, molecular detection and typing was performed on papilloma samples from eight hair goats with teat papillomatosis. The papilloma samples were tested with degenerate (FAP59/FAP64,MY09/MY11) and type-specific primers. One sample was positive with degenerate (MY09/MY11) and two samples were positive with Caprahircus papillomavirus-1 type specific primers. The MY09/MY11 primer sequence indicated identity to the closest Ovine papillomavirus type-2 (77.9%). The ChPV-1 primer sequence was similar to the closest ChPV-1 (99.4%). Investigating papillomavirus types in different animal species is important for determining the evolution, prevalence, host range, and interspecies transmission potential of papillomaviruses, and to indicate suitable therapeutics for later development.

Loop diuretics improve conditions of dialysis inception in advanced CKD: an observational cohort study.

BACKGROUND: Diuretics can reduce fluid overload but their effects on conditions of dialysis start remain elusive. We aimed to determine whether loop diuretics exposure in the year before inception can delay the need for dialysis, affect the conditions of dialysis start, and cause early mortality three months after initiation in pre-dialysis patients. METHODS: All adult patients starting dialysis from 2009 to 2015 in the REIN registry were included. Three subgroups were defined according to diuretics exposure: "continuous", "stopped", or "no diuretics" over the year before inception and compared for pre-dialysis hospitalization rates, and 3-month mortality after dialysis. RESULTS: Among 59,302 patients, we found fewer emergency initiations of dialysis in the continuous diuretics group than in the stopped diuretics and no diuretics groups: 9492 (27.5%) vs 1905 (32.3%) and 5226 (35.0%), respectively; p < 0.0001. In the continuous diuretics group, there were fewer starts on central venous catheters than in the stopped diuretics and no diuretics groups: 16,677 (49.4%) vs. 3246 (56.0%) vs. 8,639 (58.4%); p < 0.0001. Patients with continuous diuretic exposure had a lower hospitalization rate than the stopped diuretics group in the year prior to dialysis, except for heart failure. The unadjusted 3-month hazard ratio of mortality after dialysis inception was significantly higher in the "no diuretics" or "stopped diuretics" groups compared with "continuous diuretics", but the excess of risk was blunted after adjustment for emergency start and pre-dialysis visits to a nephrologist. CONCLUSION: Continuous loop diuretics exposure in the year before dialysis was associated with better conditions of dialysis inception, and possibly lower mortality rates in the three months after inception.

Serum and Milk Levels of Antibodies to Bovine Viral Diarrhoea Virus, Bovine Herpesvirus-1 and -4, and Circulation of Different Bovine Herpesvirus-4 Genotypes in Dairy Cattle with Clinical Mastitis.

Introduction: Bovine viral diarrhoea virus (BVDV) and bovine herpesvirus (BoHV)-1 and -4 are important causes of respiratory diseases and reproductive disorders of dairy cattle worldwide. Material and Methods: Investigation of BVDV and BoHV-1 and -4 antibody levels in the serum and milk of dairy cattle in a group with clinical mastitis and a healthy group was undertaken using an indirect ELISA, and identification of the BoHV-4 genotypes in clinical mastitis cases was attempted by PCR and sequencing. Results: Antibodies specific to BVDV, BoHV-1 and BoHV-4 were detected in the serum and milk of all dairy cattle with clinical mastitis. The cut-off values for BVDV and BoHV-1 in the sera and milk were extremely high in both healthy and mastitic animals. However, BoHV-4 antibodies were detected only in the clinically mastitic cattle, and BoHV-4 levels were higher in milk than in sera among these animals. Genotypes I and II of BoHV-4 were detected in the milk samples of four seropositive cows with clinical mastitis from the same herd. Conclusion: The results of this investigation demonstrate that clinical mastitis cases in the same herd may have aetiology in different BoHV-4 genotypes.

The effect of multiple-dose ivermectin treatment on CD4+/CD8+ and the oxidative stress index in goats with udder viral papillomatosis.

This study aims to reveal the therapeutic effect of ivermectin against Capra hircus papillomavirus (ChPV-1) infection and on the CD4+/CD8+ (cluster of differentiation) and oxidative stress index (OSI). Twenty hair goats naturally infected with ChPV-1 were divided into two groups with equal numbers as the ivermectin group and the control groups. Ivermectin was administered subcutaneously at a dose of 0.2 mg/kg to the goats in the ivermectin group on days 0, 7, and 21. Blood samples were collected from the vena jugularis on days 0, 21, 45, and 90. The cluster of differentiation4+/CD8+ ratio was significantly higher in the ivermectin group than in the control group on the 90th day. Furthermore, the CD8+ concentration was significantly decreased in the ivermectin group on the 90th day compared with the control group. Both total oxidant status (TOS) and OSI were found to be significantly higher in the control group on the 21st and 45th days than in the ivermectin group. On the 90th day, it was determined that the lesions in the ivermectin group improved significantly compared to those in the control group. Additionally, only in the ivermectin group was there a significant difference between the 90th day and the other days in terms of healing. As a result, it can be suggested that ivermectin has positive effects on the immune response and that its oxidative actions are of therapeutic value and do not harm the systemic oxidative status, as in untreated goats.

Prevalance of Schmallenberg orthobunyavirus (SBV) infection in sampled ruminants in Turkey's Eastern Mediterranean region between 2015 and 2017.

Schmallenberg orthobunyavirus (SBV), first discovered in 2011, belongs to the Orthobunyavirus genus of the Peribunyaviridae family. SBV, which predominantly infects ruminants, can cause severe fetal malformations when pregnant animals are infected during a critical phase of gestation. In this study, 1590 blood serum samples from cattle, sheep, and goats were obtained for serological investigation and 1604 specimens for virological investigation (including 1414 whole blood with EDTA, 165 vaginal swab samples from aborting animals, and tissue samples from 25 dead and/or aborted fetuses) in private and family-type ruminant establishments in Turkey's Eastern Mediterranean region. All the blood serum samples were tested for the presence of antibodies using ELISA, which showed SBV antibodies in 29.11% (95% CI: 26.89%-31.35%). The virological samples were tested using real-time RT-PCR for SBV nucleic acid presence, which showed 3.17% (95% CI:2.32%-4.04%) were positive. Finally, 10 different Culicoides species (a total of 29,156 Culicoides, including 16,005 females and 13,151 males) were tested to identify the vectors thought to carry infections in the region. However, no SBV nucleic acid was detected in the Culicoides pools.

SARS-CoV-2 Antibody Response After a Third Dose of the BNT162b2 Vaccine in Patients Receiving Maintenance Hemodialysis or Peritoneal Dialysis.

RATIONALE & OBJECTIVE: Recent studies showed that antibody titers after vaccination against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the dialysis population are diminished as compared with the general population, suggesting the possible value of a third booster dose. We characterized the humoral response after 3 doses of the BNT162b2 vaccine in patients treated with either maintenance hemodialysis (HD) or peritoneal dialysis (PD). STUDY DESIGN: Case series. SETTING & PARTICIPANTS: 69 French patients (38 HD and 31 PD) treated at a single center who received 3 doses of the BNT162b2 vaccine. FINDINGS: Humoral response was evaluated using plasma levels of anti-SARS-CoV-2 spike protein S1 immunoglobulin measured after the second dose and at least 3 weeks after the third dose of the BNT162b2 vaccine. Patients (median age 68 years [interquartile range (IQR), 53-76 years], 65% men) had a median anti-S1 antibody level of 284 [IQR, 83-1190] AU/mL after the second dose, and 7,554 [IQR, 2,268-11,736] AU/mL after the third dose. Three patients were nonresponders (anti-S1 antibody level < 0.8 AU/mL), and 12 were weak responders (anti-S1 antibody level 0.8-50 AU/mL) after the second vaccine dose. After the third dose, 1 of the 3 initial nonresponders produced anti-spike antibody, and all the 12 initial weak responders increased their antibody levels. Patients with a greater increase in anti-S1 antibody levels after a third dose had lower antibody levels after the second dose, and a longer time interval between the second and the third dose. Adverse events did not seem to be more common or severe after a third vaccine dose. LIMITATIONS: Observational study, small sample size. Relationship between antibody levels and clinical outcomes is not well understood. CONCLUSIONS: A third dose of the BNT162b2 vaccine substantially increased antibody levels in patients receiving maintenance dialysis and appeared to be as well tolerated as a second dose.

BVDV, BHV-1 and BLV antibodies in dromedary camels of Turkey kept without and with ruminants.

Camels are the only animals bred to sustain the tradition of wrestling in Turkey and are reared within a limited set of geographic areas. Farmers of such animals may also be engaged in ruminant breeding. The current research was aimed at documenting bovine viral diarrhoea virus (BVDV), bovine herpesvirus-1 (BHV-1), and bovine leukaemia virus (BLV) infections in sera collected from dromedary camels in four different geographical regions of Turkey during the years 2019-2021. All samples were tested for BVDV, BHV-1 and BLV antibodies as well as BVDV antigen by ELISA. Antibodies against BVDV were found in 16.8% of the camel sera tested. However, none of the camels sampled were positive in terms of BHV-1 and BLV antibodies as well as BVDV antigen. The prevalence was observed higher in the herds in which ruminants were raised in addition to camels (OR = 4.583, 95% CI, 1.298-16.182), (p = 0.018), while the prevalence was observed lower in the herds in which only camels were raised. This study showed that BVDV infection was more prevalent than BHV-1 and BLV infections in Turkish dromedary camels. Herewith, the camels, being a susceptible species to numerous viral ruminant diseases, may also serve as an important source of BVDV infection for other ruminant animals in the same flock.

Development of a BoHV-4 viral vector expressing tgD of BoHV-1 and evaluation of its immunogenicity in mouse model.

In recent years, Bovine herpesvirus 4 (BoHV-4) has emerged as an attractive gene delivery viral vector, mainly for vaccination purposes in the veterinary field. In the present study, a new infectious clone of the BoHV-4 genome carrying a bacterial artificial chromosome vector (BoHV-4-BAC) was developed by homologous recombination in mammalian cell culture and bacterial systems, and exploited to express a truncated form of glycoprotein D (tgD) of Bovine herpesvirus 1 (BoHV-1) (BoHV-4-tgD∆TK) as a vaccine candidate. This construct's immunogenicity was compared to a DNA vector expressing the same antigen (pC-tgD) in a BALB/c mouse model. After the mice were immunized, total and specific antibody responses, cytokine responses, total splenocyte cells proliferation/cytotoxicity, and virus neutralization assays were conducted to analyze the immune response elicited by both constructs. Mice from both vaccine groups developed significant humoral and cellular immune responses after a booster dose regime was conducted on day 28 post-injection. In almost all immunological assays, BoHV-4-tgDΔTK induced as high an immune response as pC-tgD. In both vaccine constructs, neutralizing antibodies were a significant determining factor in protection against BoHV-1, even after the first injection. We conclude that a BoHV-4-based viral vector offers an effective immunization strategy as an alternative to DNA-based immunization platforms, at least to combat BoHV-1.

Genetic variability of bovine herpesvirus type 4 (BoHV-4) field strains from Turkish cattle herds.

Bovine herpesvirus type 4 (BoHV­4) is a common virus in the world that is detected in clinically ill or in apparently healthy cattle. This study provides a molecular characterization of BoHV­4 strains from 24 cattle some showing respiratory and/or reproductive problems and some without any apparent clinical sign. This study also reported the growth properties of five BoHV­4 field isolates. The 24 sampled cattle came from 13 different herds in 10 provinces collected between 2007 and 2018. Phylogenetic analysis using partially amplified nucleotide sequences of ORF8 genes coding glycoprotein B (n = 24) and ORF3 genes coding thymidine kinase (n = 9), demonstrated genetic variability among the BoHV­4 strains analysed. The partial gB gene sequences clustered in three different genotypes (genotype I, II and III) were located within the genotype I cluster, such as Movar strain. The analysis of the five BoHV­4 strains isolated from vaginal swabs (n = 2), nasal swab (n = 1), and brain samples (n = 2) revealed no significant differences in their growth properties in MDBK cell culture.

Akabane virus infection in Eastern Mediterranean Region in Turkey: Culicoides (Diptera: Ceratopogonidae) as a possible vector.

Akabane virus (AKAV), which causes Akabane disease, is an arthropod-borne virus (arbovirus) transmitted by Culicoides biting midges and mosquitoes. AKAV is an important pathogen that causes abortion and congenital anomalies in ruminants. In this study, we determined the prevalence of AKAV infection and identified possible viral vectors in Turkey's Eastern Mediterranean region. The presence and prevalence of AKAV infection were assessed using serological and virological methods. Serologically, the prevalence of AKAV antibodies in cattle, sheep and goats were 44.74% (400/894), 22.90% (60/262) and 14.52% (63/434), respectively, while the total prevalence was 32.89% (523/1590). AKAV-specific nucleic acid amplicons were obtained by real-time RT-PCR from 1.13% (9/799) and 1.74% (5/288) of the cattle and sheep tested, respectively. No goats were positive for AKAV RNA. Overall, AKAV-specific nucleic acid amplicons were detected in 0.87% (14/1604) of the sampled ruminants. In addition, specimens of the assumed vector, Culicoides, were caught using light traps and identified. Ten Culicoides species were detected in the area, of which Culicoides schultzei complex was the dominant species although 32 specimens could not be identified at the species level. These were defined as Culicoides spp. AKAV nucleic acid was detected in C. schultzei, Culicoides longipennis and Culicoides circumscriptus. Phylogenetic analysis indicated two different AKAV genogroups (genogroups Ib and genogroups II) while potential AKAV vectors in this region are C. schultzei complex, C. longipennis and C. circumscriptus.

Biomarkers of Cytokine Release Syndrome Predict Disease Severity and Mortality From COVID-19 in Kidney Transplant Recipients.

BACKGROUND: Data on coronavirus disease 2019 (COVID-19) in immunocompromised kidney transplant recipients (KTR) remain scanty. Although markers of inflammation, cardiac injury, and coagulopathy have been previously associated with mortality in the general population of patients with COVID-19, their prognostic impact amongst KTR with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has not been specifically investigated. METHODS: We conducted a cohort study of 49 KTR who presented with COVID-19. Clinical and laboratory risk factors for severe disease and mortality were prospectively collected and analyzed with respect to outcomes. The study participants were divided into 3 groups: (1) mild disease manageable in an outpatient setting (n = 8), (2) nonsevere disease requiring hospitalization (n = 21), and (3) severe disease (n = 20). RESULTS: Gastrointestinal manifestations were common at diagnosis. The 30-day mortality rate in hospitalized patients was 19.5%. Early elevations of C-reactive protein (>100 mg/L) and interleukin-6 (>65 ng/L) followed by increases in high-sensitivity troponin I (>30 ng/L) and D-dimer (>960 ng/mL) were significantly associated with severe disease and mortality. Viral load did not have prognostic significance in our sample, suggesting that outcomes were chiefly driven by a cytokine release syndrome (CRS). CONCLUSIONS: Regular monitoring of CRS biomarkers in KTR with COVID-19 is paramount to improve clinical outcomes.

Assessment of replication of bovine herpesvirus type 4 in human glioblastoma and breast cancer cells as a potential oncolytic virus.

Oncolytic viruses have been extensively used in cancer treatment due to their tropism, selective replication only in tumor cells, and possible synergic interaction with other therapeutics. Different researchers have demonstrated that bovine herpesvirus 4 (BoHV-4), a member of the gammaherpesviridae family, has oncolytic potential in some human-origin cancer cell lines like glioma through the selective replication strategy. Using four apoptosis detection methods, namely MTT, LDH, TUNEL, and Annexin V assays, we evaluated the apoptotic effect of BoHV-4 Movar33/63 reference strain along with a recombinant BoHV-4 expressing EGFP in U87 MG cells (human glioblastoma cell line), MDA MB-231 (human breast cancer cell line), and MCF10a (non-tumorigenic human mammary epithelial cell line). Our findings indicate that this virus can replicate and induce apoptosis in these cell lines and hinder in vitro proliferation in a dose-dependent manner. In conclusion, BoHV-4 has in vitro potential as a novel oncolytic virus in human cancer therapy. However, its replication potential in the MCF10a cells as a non-tumorigenic human mammary epithelial cell line is a concern in using this virus in cancer therapy, at least against human mammary tumors. Further studies must therefore be conducted to examine the specific apoptotic pathways induced by this virus to move on to further experiments.

Genomic and phylogenetic characterization of ChPV2, a novel goat PV closely related to the Xi-PV1 species infecting bovines.

BACKGROUND: Papillomaviruses (PVs) infecting artiodactyls are very diverse, and only second in number to PVs infecting primates. PVs associated to lesions in economically important ruminant species have been isolated from cattle and sheep. METHODS: Potential PV DNA from teat lesions of a Damascus goat was isolated, cloned and sequenced. The PV genome was analyzed using bioinformatics approaches to detect open reading frames and to predict potential features of encoded proteins as well as putative regulatory elements. Sequence comparison and phylogenetic analyses using the concatenated E1E2L2L1 nucleotide and amino acid alignments was used to reveal the relationship of the new PV to the known PV diversity and its closest relevants. RESULTS: We isolated and characterized the full-genome of novel Capra hircus papillomavirus. We identified the E6, E7, E1, E2, L2, L1 open reading frames with protein coding potential and putative active elements in the ChPV2 proteins and putative regulatory genome elements. Sequence similarities of L1 and phylogenetic analyses using concatenated E1E2L2L1 nucleotide and amino acid alignments suggest the classification as a new PV type designated ChPV2 with a phylogenetic position within the XiPV genus, basal to the XiPV1 species. ChPV2 is not closely related to ChPV1, the other known goat PV isolated from healthy skin, although both of them belong confidently into a clade composed of PVs infecting cervids and bovids. Interestingly, ChPV2 contains an E6 open reading frame whereas all closely related PVs do not CONCLUSION: ChPV2 is a novel goat PV closely related to the Xi-PV1 species infecting bovines. Phylogenetic relationships and genome architecture of ChPV2 and closely related PV types suggest at least two independent E6 losses within the XiPV clade.

In-depth virological assessment of kidney transplant recipients with COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread widely, causing coronavirus disease 2019 (COVID-19) and significant mortality. However, data on viral loads and antibody kinetics in immunocompromised populations are lacking. We aimed to determine nasopharyngeal and plasma viral loads via reverse transcription-polymerase chain reaction and SARS-CoV-2 serology via enzyme-linked immunosorbent assay and study their association with severe forms of COVID-19 and death in kidney transplant recipients. In this study, we examined hospitalized kidney transplant recipients with nonsevere (n = 21) and severe (n = 19) COVID-19. SARS-CoV-2 nasopharyngeal and plasma viral load and serological response were evaluated based on outcomes and disease severity. Ten recipients (25%) displayed persistent viral shedding 30 days after symptom onset. The SARS-CoV-2 viral load of the upper respiratory tract was not associated with severe COVID-19, whereas the plasma viral load was associated with COVID-19 severity (P = .010) and mortality (P = .010). All patients harbored antibodies during the second week after symptom onset that persisted for 2 months. We conclude that plasma viral load is associated with COVID-19 morbidity and mortality, whereas nasopharyngeal viral load is not. SARS-CoV-2 shedding is prolonged in kidney transplant recipients and the humoral response to SARS-CoV-2 does not show significant impairment in this series of transplant recipients.

Detection of genotype 1 bovine leukemia virus from a C.schultzei pool: Do Culicoides spp. have a role on the transmission of bovine leukemia virus?

Bovine leukemia virus (BLV) is known as the etiological agent of Enzootic bovine leukosis (EBL), which is the most common neoplastic disease of cattle. While the major route of virus transmission is believed to be iatrogenic, BLV proviral DNA has been identified in biological materials, including nasal secretions, saliva, milk, colostrum, and semen, and in several insect species, including horses flies. However, insects' role in the natural transmission of BLV has not been clearly demonstrated. This study assessed the possible role of midges - Culicoides spp. - in BLV transmission. BLVs were genetically characterized and BLV infection seroprevelance was determined in 224 cattle sampled from 27 different small family herds in five different districts in Hatay province, southern Turkey. Out of the 25 Culicoides spp. pools, one (4.0%; 1/25) was a C.schultzei pool while 2.67% (6/224) of the sampled cattle were positive for BLV nucleic acid. The seroprevalance rates for the sampled herds and all sampled cattle were 7.40% (2/27) and 1.33% (3/224), respectively. According to the phylogenetic analysis, the sequences of the BLVs from the cattle (n = 6) and the one BLV-positive C.schultzei pool clustered on genotype 1 (G1) BLVs. Although these results do not reveal the exact role of Culicoides spp. or other midges flies in BLV transmission, the simultaneous presence of same substitions in BLVs from both cattle and a C.schultzei pool is noteworthy. Further studies on the env gene and other BLV gene regions detected from cattle and C.schultzei pools are ongoing to understand the possible epidemiological relationship between cattle and flies.