Evidences of HEV genotype 3 persistence and reactivity in liver parenchyma from experimentally infected cynomolgus monkeys (Macaca fascicularis).

Hepatitis E virus genotype 3 (HEV-3) is an emerging zoonotic pathogen, responsible for sporadic cases of acute hepatitis E worldwide. Primate models have proven to be an essential tool for the study of HEV pathogenesis. Here we describe the outcomes of HEV infection in Macaca fascicularis (cynomolgus) inoculated experimentally with genotype 3. Eight adult cynomolgus macaques were inoculated intravenously with HEV-3 viral particles isolated from swine and human samples. Liver, spleen, duodenum, gallbladder and bile were sequential assessed up to the end-point of this study, 67 days post-inoculation (dpi). Our previously published findings showed that biochemical parameters return gradually to baseline levels at 55 dpi, whereas anti-HEV IgM and HEV RNA become undetectable in the serum and feces of all animals, indicating a non-viremic phase of recovery. Nevertheless, at a later stage during convalescence (67 dpi), the presence of HEV-3 RNA and antigen persist in central organs, even after peripheral viral clearance. Our results show that two cynomolgus inoculated with swine HEV-3 (animals I3 and O1) presented persistence of HEV RNA low titers in liver, gallbladder and bile. At this same stage of infection, HEV antigen (HEV Ag) could be detected in all infected animals, predominantly in non-reactive Kupffer cells (CD68+iNOS-) and sinusoidal lining cells. Simultaneously, CD4+, CD3+CD4+, and CD3+CD8+ immune cells were identified in hepatic sinusoids and small inflammatory clusters of lobular mononuclear cells, at the end-point of this study. Inability of HEV clearance in humans can result in chronic hepatitis, liver cirrhosis, with subsequent liver failure requiring transplantation. The results of our study support the persistence of HEV-3 during convalescence at 67 dpi, with active immune response in NHP. We alert to the inherent risk of viral transmission through liver transplantation, even in the absence of clinical and biochemical signs of acute infection. Thus, besides checking conventional serological markers of HEV infection, we strongly recommend HEV-3 RNA and antigen detection in liver explants as public health measure to prevent donor-recipient transmission and spread of hepatitis E.

Acute acalculous cholecystitis in children.

Acute acalculous cholecystitis (AAC) is the inflammatory disease of the gallbladder in the absence of gallstones. AAC is estimated to represent at least 50% to 70% of all cases of acute cholecystitis during childhood. Although this pathology was originally described in critically ill or post-surgical patients, most pediatric cases have been observed during several infectious diseases. In addition to cases caused by bacterial and parasitic infections, most pediatric reports after 2000 described children developing AAC during viral illnesses (such as Epstein-Barr virus and hepatitis A virus infections). Moreover, some pediatric cases have been associated with several underlying chronic diseases and, in particular, with immune-mediated disorders. Here, we review the epidemiological aspects of pediatric AAC, and we discuss etiology, pathophysiology and clinical management, according to the cases reported in the medical literature.

[European outbreak of Hepatitis A in Iceland in 2017. Common radiological changes of the gallbladder].

IAim The incidence of hepatitis A (HAV) in Iceland is low with about one case per year in the last decades. Since 2016, there has been an ongoing outbreak of HAV in men who have sex with men (MSM). The aim of this study was to inves-tigate whether cases diagnosed in Iceland during 2017 had any link to the HAV outbreak in Europe. Methods All cases of HAV in Iceland during 2017 were reviewed retrospectively. Results Four of five cases diagnosed during 2017 were MSM and one was a female. Three cases presented the same week in the summer 2017. The age of the patients was between 25 and 39 years. All the male patients had had sex with men from Europe and/or had travelled to Europe prior to admission. All cases had typical signs and symptoms of HAV infection and in all cases recent infection was confirmed by positive serology and exclusion of other causes of acute hepatitis. Four of five patients had radiological signs of changes in the gallbladder with thickening of the wall and oedema and one underwent later an elective cholecystectomy. Conclusion The outbreak of HAV in MSM Europe reached Iceland in the summer 2017, emphasizing the importance of vaccination in this risk group as recommended by the Icelandic Health Authorities. The review of these cases indicate that changes such as thickening of the gallbladder wall without gallstones in patients with HAV are common. It is important to discrimi-nate patients with these changes associated with HAV from patients with acute acalculus cholecystitis.

Roles of VP4 and NSP1 in determining the distinctive replication capacities of simian rotavirus RRV and bovine rotavirus UK in the mouse biliary tract.

Rotavirus replication and virulence are strongly influenced by virus strain and host species. The rotavirus proteins VP3, VP4, VP7, NSP1, and NSP4 have all been implicated in strain and species restriction of replication; however, the mechanisms have not been fully determined. Simian (RRV) and bovine (UK) rotaviruses have distinctive replication capacities in mouse extraintestinal organs such as the biliary tract. Using reassortants between UK and RRV, we previously demonstrated that the differential replication of these viruses in mouse embryonic fibroblasts is determined by the respective NSP1 proteins, which differ substantially in their abilities to degrade interferon (IFN) regulatory factor 3 (IRF3) and suppress the type I IFN response. In this study, we used an in vivo model of rotavirus infection of mouse gallbladder with UK × RRV reassortants to study the genetic and mechanistic basis of systemic rotavirus replication. We found that the low-replication phenotype of UK in biliary tissues was conferred by UK VP4 and that the high-replication phenotype of RRV was conferred by RRV VP4 and NSP1. Viruses with RRV VP4 entered cultured mouse cholangiocytes more efficiently than did those with UK VP4. Reassortants with RRV VP4 and UK NSP1 genes induced high levels of expression of IRF3-dependent p54 in biliary tissues, and their replication was increased 3-fold in IFN-α/ß and -γ receptor or STAT1 knockout (KO) mice compared to wild-type mice. Our data indicate that systemic rotavirus strain-specific replication in the murine biliary tract is determined by both viral entry mediated by VP4 and viral antagonism of the host innate immune response mediated by NSP1.

Pathological studies on postvaccinal reactions of Rift Valley fever in goats.

RVF live attenuated vaccine (Smithburn strain) was evaluated by using goats as experimental animal. The results indicate that this vaccine cause severe deleterious pathological changes in liver especially in kids and causing abortion in pregnant does. The virus was seen to be propagated inside hepatic cells forming intranuclear inclusions which was also seen by E.M. Viral antigens were detected in hepatic cells, gall bladder, endothelial lining of blood vessels, leukocytes, kidneys and heart by using immunoflourescent technique. It could be concluded that the use of live attenuated vaccine of RVF (Smithburn strain) for immunization of live stock is not safe in Egypt as it considered an endemic area.

Acute cytomegalovirus cholecystitis following renal transplantation.

Solid organ transplant recipients are at risk of infection from cytomegalovirus (CMV). A wide range of disease is associated with CMV infection and we report two cases of CMV cholecystitis in patients following renal transplantation. Both patients presented with severe hemorrhagic cholecystitis, which required immediate resuscitation and emergency cholecystectomy. The diagnosis of CMV infection was confirmed in both cases using CMV-specific staining of the gallbladder. The diagnosis of CMV cholecystitis must be considered in all patients with upper abdominal pain after renal transplantation.

Detection of bovine viral diarrhoea virus (BVDV) nucleic acid and antigen in different organs of water buffaloes (Bubalus bubalis).

Bovine viral diarrhoea virus (BVDV) is a pestivirus that infects mainly bovine cattle. Nevertheless, there are several reports about infections in other members of the Artiodactyla order including serological studies, that indicate infection of BVDV in buffaloes. The aim of this article is to study the presence of BVDV in three young water buffaloes, displaying nonspecific clinical signs, compatible with the BVDV infection. Both immunohistochemistry and RT-PCR confirmed the presence of BVDV in the animals. The sequence analysis on RT-PCR amplicons revealed high identity with reference strains of genotypes 1a and 1b. Although BVDV was unequivocally identified in the sick animals, it has not been proved it is responsible for the clinical signs. Further studies are needed to clarify the pathogenic role of BVDV infection in this animal species, and the role of buffaloes in the epidemiology of BVDV infection.

[Changes of pH and kinematic viscosity of the contents of gall-bladder at the pre-gallstone stage of cholelithiasis].

The aim of the work was to study the indicators of pH and kinematic viscosity of the contents of the gall-bladder at receiving B-portion and their changes after a 7-hour period according to the biliary sediment of patients and to establish their link to the infection. Total 31 patients aged 25-55 with biliary sediment in lumens of their gall-bladders (a general group) and 5 healthy patients (a control group) were studied by us. In all cases the markers of the virus of Hepatitis B in blood serum were determined and ultrasonic examination of gall-bladder was carried out by us prior to the duodenal intubation. B-portion was subjected to bacteriological research for aerobes and anaerobs, and pH and kinematic viscosity (eta=mm(2)/sec) were determined. The patients from the control as well as from the general group after a 7-hour period were redetermined pH and Kinematic viscosity of B-portion. Kinematic viscosity was determined with a capillary viscosimeter, and pH was fixed by a method of potentiometer. Statistical treatment was undertaken by the method of ANOLA p<0,05. During the duodenal intubation the mean value of pH within the control group was fixed at 6.74, and the mean value of the kinematic viscosity was 1.34, and after a 7-hour period the same indicators constituted 6.87 and 1.35 mm(2)/sec, respectively. The same indicators within the general group during duodenal intubation were the following: 7.26 and 1.99 mm(2)/sec, and after a 7-hour period the same indicators were 7.78 and 2.19 mm(2)/sec, respectively. There was a significant statistical difference between the mean values of pH and kinematic viscosity of the contents of gall-bladder of the patients from the control and general groups. During a 7-hour period, there was also a significant statistical difference between the changes of the same indicators. The difference was even greater in case of infection. Following the above-mentioned we can conclude that in case of sediment in the lumen of gall-bladder, inclination of pH to alkalinity and increase in kinematic viscosity of the contents is one of the most important criteria of lithogenicity of gall-bladder contents, and one of the reasons for such changes is chronic infection of a gall-bladder.

Immunohistochemical studies of productive rhesus cytomegalovirus infection in rhesus monkeys (Macaca mulatta) infected with simian immunodeficiency virus.

In humans infected with the human immunodeficiency virus (HIV), clinical disease due to human cytomegalovirus (HCMV) infection is one of the AIDS-defining diseases; HCMV is the most common opportunistic infection found postmortem. Histologically, the typical lesions are characterized by "owl's eye cells." In rhesus monkeys infected with simian immunodeficiency virus (SIV), comparable lesions are caused by an infection with the rhesus CMV (RhCMV). The aim of this study was to investigate the incidence of productive and latent RhCMV infection in monkeys infected with SIV macaques (SIVmac). Eleven SIVmac-infected rhesus monkeys, which were euthanatized after developing AIDS-like disease, and 11 clinically healthy and uninfected animals comprised the study. The monkeys were screened serologically for RhCMV by western-blot analysis. Immunohistochemistry was performed by an indirect immunoperoxidase technique with a polyclonal rabbit RhCMV-antiserum. Lesions characteristic of RhCMV-associated diseases were detected histologically. All animals were latently RhCMV-infected. Seven of eleven (63.6%) SIV-infected macaques were productively RhCMV infected according to immunohistochemistry. RhCMV antigen was identified in the gastrointestinal tract, the hepatobiliary system, the lungs, and the testicles. Two of these seven animals showed characteristic inflammatory lesions associated with productive infection. Consequently, the CMV prevalence in SIVmac-infected rhesus monkeys and human AIDS patients is comparable.

PCR studies on the potential sites for latency of BHV-4 in calves.

The aim of the study was to examine various tissues of experimentally infected calves for the BHV-4 genome so as to detect in which cells the virus persists during the latent phase of the infection. The presence of the bovine herpesvirus type 4 genome was detected by a nested PCR in a variety of tissues collected from two susceptible calves experimentally infected 62 days earlier. Mild clinical signs of bronchitis, an elevated body temperature for 2-3 days, and a slightly increased number of blood leukocytes were observed in both inoculated calves. BHV-4 was demonstrated in seven samples from the 12 different parts of the nervous system tested from each calf (29.1%), from the cornea, from lymph nodes near to the inoculation site, from the gallbladder and from the bone marrow. Thus a member of the predominantly lymphotropic Gammaherpesvirinae subfamily was detected in neural tissue and other organs that have never been associated with persistence.

[PCR in detecting the correlation between infection of HBV and cholelithiasis].

OBJECTIVE: To study the correlation between cholelithiasis and the infection of HBV. METHOD: 32 formalin-fixed and paraffin-embedded gallbladder samples of cholelithiasis patients and 20 gallbladder samples of non-cholelithiasis patients were investigated using the polymerase chain reaction-Ethidium bromide (PCR-EB) assay. The 52 patients were positive to HBV serologic markers. RESULT: The results showed that HBV-DNA was found in 13 gallbladder samples of 32 cholelithiasis patients (40.63%), significantly higher than that in 3 gallbladder samples of 20 non-cholelithiasis patients (15%). CONCLUSION: The infection of HBV and the formation of cholelithiasis are correlated.

Expression of human leukocyte antigens class I and class II on cultured biliary epithelial cells after cytomegalovirus infection.

The influence of cytomegalovirus (CMV) infection on the HLA expression on cultured biliary epithelial cells (BEC) was investigated. CMV-infection augmented expression of HLA class I but not of HLA class II. CMV reduced the IFN-gamma-mediated induction of the de novo expression of HLA class II while the stimulated expression of HLA class I was not impaired. Autologous but not allogeneic PBL responded to CMV-infected BEC. This response resulted in upregulation of HLA class I on BEC which was significantly higher compared with the expression on infected BEC alone or on uninfected BEC cocultured with autologous PBL. The results suggest that CMV modulates the immunogenic potential of BEC, which is important for the HLA and CMV-mediated pathomechanisms in vivo.