Structural similarity of human papillomavirus E4 and polyomaviral VP4 exhibited by genomic analysis of the common kestrel (Falco tinnunculus) polyomavirus.

Polyomaviruses are widely distributed viruses of birds that may induce developmental deformities and internal organ disorders primarily in nestlings. In this study, polyomavirus sequence was detected in kidney and liver samples of a common kestrel (Falco tinnunculus) that succumbed at a rescue station in Hungary. The amplified 5025 nucleotide (nt) long genome contained the early (large and small T antigen, LTA and STA) and late (viral proteins, VP1, VP2, VP3) open reading frames (ORFs) typical for polyomaviruses. One of the additional putative ORFs (named VP4) showed identical localization with the VP4 and ORF-X of gammapolyomaviruses, but putative splicing sites could not be found in its sequence. Interestingly, the predicted 123 amino acid (aa) long protein sequence showed the highest similarity with human papillomavirus E4 early proteins in respect of the aa distribution and motif arrangement implying similar functions. The LTA of the kestrel polyomavirus shared <59.2% nt and aa pairwise identity with the LTA sequence of other polyomaviruses and formed a separated branch in the phylogenetic tree among gammapolyomaviruses. Accordingly, the kestrel polyomavirus may be the first member of a novel species within the Gammapolyomavirus genus, tentatively named Gammapolyomavirus faltin.

Recombinant Lloviu virus as a tool to study viral replication and host responses.

Next generation sequencing has revealed the presence of numerous RNA viruses in animal reservoir hosts, including many closely related to known human pathogens. Despite their zoonotic potential, most of these viruses remain understudied due to not yet being cultured. While reverse genetic systems can facilitate virus rescue, this is often hindered by missing viral genome ends. A prime example is Lloviu virus (LLOV), an uncultured filovirus that is closely related to the highly pathogenic Ebola virus. Using minigenome systems, we complemented the missing LLOV genomic ends and identified cis-acting elements required for LLOV replication that were lacking in the published sequence. We leveraged these data to generate recombinant full-length LLOV clones and rescue infectious virus. Similar to other filoviruses, recombinant LLOV (rLLOV) forms filamentous virions and induces the formation of characteristic inclusions in the cytoplasm of the infected cells, as shown by electron microscopy. Known target cells of Ebola virus, including macrophages and hepatocytes, are permissive to rLLOV infection, suggesting that humans could be potential hosts. However, inflammatory responses in human macrophages, a hallmark of Ebola virus disease, are not induced by rLLOV. Additional tropism testing identified pneumocytes as capable of robust rLLOV and Ebola virus infection. We also used rLLOV to test antivirals targeting multiple facets of the replication cycle. Rescue of uncultured viruses of pathogenic concern represents a valuable tool in our arsenal for pandemic preparedness.

Companion animals likely do not spread COVID-19 but may get infected themselves.

Coronavirus disease 2019 (COVID-19) is a highly contagious infectious disease caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). From the epidemiological data, the picture emerges that the more severe etiopathologies among COVID-19 patients are found in elderly people. The risk of death due to COVID-19 increases exponentially with age. Eight out of 10 COVID-19 related deaths occur in people older than 65 years of age. Older patients with comorbid conditions such as hypertension, heart failure, diabetes mellitus, asthma, chronic obstructive pulmonary disease, and cancer have a much higher case fatality rate. Governments and public health authorities all over the world have realized that protections of vulnerable older adults should be a priority during the COVID-19 pandemic. COVID-19 is a zoonotic disease. The SARS-CoV-2 virus was originally transmitted likely from a bat or a pangolin to humans. Recent evidence suggests that SARS-CoV-2, similar to other coronaviruses, can infect several species of animals, including companion animals such as dogs, cats, and ferrets although their viral loads remain low. While the main source of infection transmission therefore is human to human, there are a few rare cases of pets contracting the infection from a SARS-CoV-2-infected human. Although there is no evidence that pets actively transmit SARS-CoV-2 via animal-to-human transmission, senior pet ownership potentially may pose a small risk to older adults by (1) potentially enabling animal-to-human transmission of SARS-CoV-2 in the most vulnerable population and (2) by increasing the exposition risk for the elderly due to the necessity to care for the pet and, in the case of dogs, to take them outside the house several times per day. In this overview, the available evidence on SARS-CoV-2 infection in pets is considered and the potential for spread of COVID-19 from companion animals to older individuals and the importance of prevention are discussed.

Crimean-Congo hemorrhagic fever virus infection triggers the upregulation of the Wnt signaling pathway inhibitor genes.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a highly pathogenic agent. Thus far, vaccines and specific antiviral therapies are not available against the threat of infection. Our knowledge regarding its pathogenesis is indeed limited, and thus, developing effective antiviral therapies is hampered. Several studies have demonstrated that the CCHFV infection has an impact on numerous signal transduction pathways. In parallel, the Wnt signaling pathway components are responsible for different important biological processes including cell fate determination, cell migration and cell polarity. Moreover, its implication among several virus infections has been proven, yet little is known in reference to which components of the Wnt pathway are being activated/inhibited as a response to the infection. Our aim was to elicit the influence of the CCHFV infection on adenocarcinomic human alveolar basal epithelial cells in vitro regarding the Wnt signaling pathway-related genes. Gene-expression changes of 92 Wnt-associated genes were examined 48 h post-infection. Furthermore, ß-catenin levels were compared in the infected and uninfected cells. Significant changes were observed in the case of 13 genes. The majority of the upregulated genes are associated with the inhibition of the Wnt/ß-catenin signaling. Additionally, infected cells expressed less ß-catenin. Our findings suggest that CCHFV blocks the Wnt/ß-catenin pathway. Our study corroborates the link between CCHFV infection and the Wnt signaling pathways. In addition, it broadens our knowledge in the CCHFV pathomechanism.

[Milestones in liver surgery]. / Mérföldkövek a máj sebészetében.

Numerous outstanding summarizing publications on the milestones of the rapid development of liver surgery in the twentieth century came to light around the year 2000, therefore in this summary only the newest principles and novelties of liver surgery after the second millennium are presented. Among the new principles, the newest indications, the bloodless surgery and the vascular exclusion of the liver, the "associating liver partition and portal vein occlusion for staged hepatectomy" (ALPPS) - as presently the fastest and most successful method for liver regeneration, and the "downsizing of tumors" treatment for the interest of resection of unresectable tumours are analyzed and evaluated. Open, laparoscopic, and robotic liver surgery are discussed by comparing blood loss, operating time, cost, and long-term results. Laparoscopic liver surgery on the basis of evidences has become "the method of choice" in our days. Robotic liver surgery needs further evidence-based data for determination of its place in the clinical practice. An intention is also composed in terms of place-determination of liver surgery between gastrointestinal, hepatopancreatobiliary, liver transplantation, and oncologic surgery. The mortality rate has decreased to 1% recently; the data regarding morbidity are variable; the modified Clavien-Dindo, the modified Accordion complication system and the comprehensive complication index are intended to move the different evaluations to a common ground. The up-to-date idea of hepatic surgeon, liver center and liver surgery are defined in close cooperation with international theoretical and practical outcome, in the limelight of multidisciplinarity and multimodality. The internationally observed inequalities in liver surgery are also discussed from the point of view of tackling with the inequalities existing in the universal healthcare systems on a local, national and global level by collecting and controlling the results systematically, and developing and implying international guidelines on the basis of evidences. Orv Hetil. 2018; 159(10): 375-383.

Down-regulation of Toll-like Receptor TLR4 Is Associated with HPV DNA Integration in Penile Carcinoma.

BACKGROUND/AIM: Development of penile cancers is attributed to HPV-related carcinogenesis. Our aim was to analyze HPV positivity and TLR4, p16ink4a and p53 expression. MATERIALS AND METHODS: HPV presence was assessed with virus-specific TaqMan PCR and HPV Genotyping Test in 31 penile cancers. Immunohistochemistry was carried out on tissue microarray. RESULTS: TLR4 expression was detected in 4 of the 16 HPV positive and 13 of the 15 HPV negative tumors. We found a significant inverse correlation between HPV positivity and TLR4 expression (p=0.0006). Ten of the 16 HPV-positive but none of the 15 HPV-negative tumors expressed p16INK4a. A significant correlation was seen between p53 expression and lack of HPV DNA (p=0.0191) as well as between TLR4 and p53 expression (p=0.0198) in penile cancers. CONCLUSION: Our findings suggest a protective role of TLR4 expression against HPV DNA integration and the viral and non-viral carcinogenesis of penile cancer.

Decreased functional activity of multidrug resistance protein in primary colorectal cancer.

BACKGROUND: The ATP-Binding Cassette (ABC)-transporter MultiDrug Resistance Protein 1 (MDR1) and Multidrug Resistance Related Protein 1 (MRP1) are expressed on the surface of enterocytes, which has led to the belief that these high capacity transporters are responsible for modulating chemosensitvity of colorectal cancer. Several immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) studies have provided controversial results in regards to the expression levels of these two ABC-transporters in colorectal cancer. Our study was designed to determine the yet uninvestigated functional activity of MDR1 and MRP1 transporters in normal human enterocytes compared to colorectal cancer cells from surgical biopsies. METHODS: 100 colorectal cancer and 28 adjacent healthy mucosa samples were obtained by intraoperative surgical sampling. Activity of MDR1 and MRP1 of viable epithelial and cancer cells were determined separately with the modified calcein-assay for multidrug resistance activity and sufficient data of 73 cancer and 11 healthy mucosa was analyzed statistically. RESULTS: Significantly decreased mean MDR1 activity was found in primary colorectal cancer samples compared to normal mucosa, while mean MRP1 activity showed no significant change. Functional activity was not affected by gender, age, stage or grade and localization of the tumor. CONCLUSION: We found lower MDR activity in cancer cells versus adjacent, apparently, healthy control tissue, thus, contrary to general belief, MDR activity seems not to play a major role in primary drug resistance, but might rather explain preferential/selective activity of Irinotecan and/or Oxaliplatin. Still, this picture might be more complex since chemotherapy by itself might alter MDR activity, and furthermore, today limited data is available about MDR activity of cancer stem cells in colorectal cancers. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1675739129145824.

Sequencing and phylogenetic analysis identifies candidate members of a new picornavirus genus in terrestrial tortoise species.

Near-complete genome sequences of seven picornavirus (PV) strains isolated from different terrestrial tortoise species were determined and characterized. The genome organization of the strains proved to be similar and displayed a typical PV layout, and the polyprotein-encoding regions showed low similarity to those of other PVs. The predicted regions of the tortoise PV genomes were related to the corresponding genome parts of viruses belonging to distinct genera, implying modular evolution of these novel viruses. Our results suggest that these tortoise PVs belong to a prototype species in a separate proposed genus in the family Picornaviridae, tentatively called Topivirus (Tortoise picornavirus).

[Surgical risk in patients with liver disease]. / Sebészi kockázat májbetegeken.

The author draws attention on the importance surgical risk analysis from patient's safety point of view. Recently the development in quality assurance affected surgical practice as well, hence determination and evaluation of surgical risk are more exactly defined. This resulted in a significant decrease in mortality during surgical interventions on the liver despite a wider indication and increased numbers, recently. Importantly, surgical risk is much higher in patients with liver disease compared to patients with normal liver. The risk of surgical interventions for liver diseases (HCC, tumor) in patients with diffuse liver diseases (cirrhosis, chronic hepatitis, ALD) can be expressed numerically. For many years the Child-Turcotte-Pugh stadium could have been determined by using actual laboratory values. Recently the "50-50 rule" or more frequently the MELD score -- originally used in the practice of liver transplantation -- mean objective expression of surgical risk. Treatment optimalisation can reduce surgical risk, selected on the basis of risk analysis in multidisciplinary settings, which focus on the need of liver surgeons.

Comparison of human hepatoma HepaRG cells with human and rat hepatocytes in uptake transport assays in order to predict a risk of drug induced hepatotoxicity.

Human hepatocytes are the gold standard for toxicological studies but they have several drawbacks, like scarce availability, high inter-individual variability, a short lifetime, which limits their applicability. The aim of our investigations was to determine, whether HepaRG cells could replace human hepatocytes in uptake experiments for toxicity studies. HepaRG is a hepatoma cell line with most hepatic functions, including a considerable expression of uptake transporters in contrast to other hepatic immortalized cell lines. We compared the effect of cholestatic drugs (bosentan, cyclosporinA, troglitazone,) and bromosulfophthalein on the uptake of taurocholate and estrone-3-sulfate in human and rat hepatocytes and HepaRG cells. The substrate uptake was significantly slower in HepaRG cells than in human hepatocytes, still, in the presence of drugs we observed a concentration dependent decrease in uptake. In all cell types, the culture time had a significant impact not only on the uptake process but on the inhibitory effect of drugs too. The most significant drug effect was measured at 4 h after seeding. Our report is among the first concerning interactions of the uptake transporters in the HepaRG, at the functional level. Results of the present study clearly show that concerning the inhibition of taurocholate uptake by cholestatic drugs, HepaRG cells are closer to human hepatocytes than rat hepatocytes. In conclusion, we demonstrated that HepaRG cells may provide a suitable tool for hepatic uptake studies.

[Latest indications for hanging manoeuvre in liver surgery]. / A hanging manover újabb indikációi a májsebészetben.

By definition the liver hanging manoeuvre (LHM) means that a slip is passed between the liver parenchyma and the inferior vena cava. It was first published by Belghiti in 200l, and several changes in the indication as well as in the method have been published since then. In parallel, the anatomical and histological basis has been clarified for LHM, too. According to general consensus LHM increases safety and radicality of liver surgery. Initially LHM was applied for removal of huge tumours infiltrating the diaphragm. Authors worked out two modifications for LHM. Tumours / primary or secondary / in segment IVA are sometimes located in close proximity to the median hepatic vein and inferior vena cava , and the resectability of these tumors can determined by the hanging manoeuvre. Tumors in segment VII can be removed by partial resection of vena cava facilitated by LHM. Four patients with LHM are discussed, and based on this limited experience as well as the latest observations from relevant literature the authors claim that LHM increases the safety of resections from segment IVA and VII. Vascular infiltration of the vena cava is always a technical challenge, which can be suspected on preoperative imaging modalities, but in borderline cases only the intraoperative ultrasound and surgical judgment together with LHM would lead to the exact diagnosis and makes the resection possible.

Binding of trastuzumab to ErbB2 is inhibited by a high pericellular density of hyaluronan.

Although trastuzumab is an efficient drug, primary and acquired resistance is a challenging problem. The authors have previously shown in mouse xenograft experiments that masking ErbB2 by hyaluronan leads to diminished binding of the antibody and consequent resistance. In the current work, they correlated trastuzumab binding with the pericellular density of hyaluronan in ErbB2-overexpressing human breast cancer samples. A method for quantifying the relative binding of trastuzumab was developed involving constant and low-frequency background subtraction, segmenting the image to membrane and background pixels followed by evaluation of trastuzumab fluorescence, normalized with the expression level of ErbB2, only in the membrane. The normalized binding of trastuzumab showed a negative correlation with the pericellular density of hyaluronan (r = -0.52) with the effect being the most pronounced in the extreme cases (i.e., low and high hyaluronan densities predicted strong and weak binding of trastuzumab, respectively). Removal of hyaluronan by hyaluronidase digestion unmasked the trastuzumab binding epitope of ErbB2 demonstrated by a significantly increased normalized binding of the antibody. The results show that the accumulation of pericellular hyaluronan plays a crucial role in masking ErbB2.

Transverse hepatectomy: a 14-years experience.

BACKGROUND/AIMS: Transverse hepatectomy, removal of hepatic segments IVB, V, VI, (III) in continuity with the gallbladder through the transverse portal plane, was first introduced by Paul Sugarbaker in 1990. In 1995 the first transverse hepatectomy in Hungary was carried out by our workgroup. This article summarizes our experience with transverse hepatectomies during the period from 1995 to 2008. METHODOLOGY: During this time 72 trisegmentectomies were performed 22 out of these operations were transverse hepatectomies. RESULTS: The average resection time for transverse hepatectomy was 1 hour and 20 minutes, there was an average 0.8 unit blood transfusion requirement per patient, the average exclusion time was 20.6 minutes, the average time spent in hospital postoperatively was 10.1 days and there were 2 perioperative deaths due to sepsis and hemorrhagic shock. CONCLUSIONS: In our experience transverse hepatectomy proved to be a bloodless, relatively easy procedure, which enables safe removal of the antero-lateral segments. This new type of major hepatic resection is very useful in metastasis surgery and has minimal associated morbidity and mortality.

[Surgical prevention of radiation enteritis: case report and review of literature]. / Irradiatiós enteritis sebészeti prevenciója: esetismertetés és az irodalom áttekintése.

Due to high morbidity and mortality rates, radiation enteritis is one of the most feared complications of abdominal and pelvic radiation therapy. Advances in radiation technology and radiation planning contributed to recent significant achievements. Surgical prevention provides further opportunities in decreasing the risk of radiation enteritis. A 75 year old male underwent transurethral resection for urothelial carcinoma of the bladder. Prior to initiation of radiation therapy, we performed Shouldice repair for bilateral inguinal hernias to prevent radiation injury to the fixed small intestines. Later our patient received 55 Gy of radiation therapy and two series of Carboplatin chemotherapy. Following radio-chemotherapy our patient did not developed radiation enteritis. In our report we discuss hernioplasty as an important method for prevention of radiation enteritis. We also review other options of surgical prevention.

[From project-based sample collection to biobank]. / Projektalapú mintagyujtéstol a biobankig.

The research group takes samples for molecular genetical examinations from tumors removed during operations within ischemic time interval. Samples are stored in liquid nitrogen. Clinical data of these patients are recorded in an informatics system developed by the group. Patients are followed in an out-patient clinic set up for this purpose not financed by the National Health Insurance Fund. Tissue samples and follow up data are used to cooperate with molecular genetical laboratories.

[Multiple hepatic abscesses due to actinomycosis: difficulties of diagnosis]. / Multiplex májtályog képében jelentkezo actinomycosis esete: a kórismézés nehézségei.

Authors present the case of a 65 year-old man without any remarkable previous illness, hospitalized because of having prolonged fever caused probably by an infection due to a tick bite. Tularaemia and bartonellosis based on serological results as well as neoplasm of the sigma with multiple metastases to the liver raised by ultrasonography and abdominal CT could be excluded with detailed investigations. Multiple abscesses of the liver were diagnosed, followed by a percutaneuos ultrasonography directed aspiration of one of the abscesses which resulted in 150 ml buffy pus. Microbiological examination evaluated the presence of Actinomyces meyeri. Instead of neoplasm, diverticulosis and diverticulitis of the sigma could be clarified with perforation of one of the diverticuli causing peridiverticulitis, retroperitoneal abscess and probably abscesses in the liver by haematogenous dissemination. Surgical intervention, resection of the liver and the sigma resulted in recovery of the patient. Difficulties of diagnosis are discussed in the article pointing out the importance of interdisciplinary collaboration, involving also infectologist. Diverticulosis of the sigma as presumably origin of hepatic actinomycosis is a literary curiosity. This was the cause of the case presentation.

Inflammation of ectopic pancreatic tissue in a Meckel's diverticulum causing acute abdominal symptoms: a case report and review of the literature.

An unusual case of acute abdomen was caused by the inflammation of ectopic pancreatic tissue in a Meckel's diverticulum. A 49-year-old man presented with acute abdominal pain, and the clinical diagnosis of acute appendicitis was established. During laparotomy, a normal appendix of unusual localization near the gallbladder and a Meckel's diverticulum with an inflamed tip were found. Histological examination showed acute inflammation of heterotopic pancreatic tissue along with normal ectopic gastric and duodenal mucosa within the wall of the diverticulum. Fat necrosis was also ascertained. The authors believe that this is the first report of acute inflammation of ectopic pancreatic tissue and the presence of normal ectopic gastric and duodenal tissue in the same Meckel's diverticulum.

[Choice of surgical strategy for colorectal liver metastases depending on clinical and pathological response to neoadjuvant chemo- and targeted therapy]. / Colorectalis májáttétek sebészeti kezelési stratégiájának megválasztása a neoadjuváns kemo- és célzott terápiára adott klinikai és patológiai válasz függvényében.

UNLABELLED: Surgical strategy of colorectal liver metastases depends on clinical and pathological response to neoadjuvant chemo- and target therapy. Recently, surgical treatment of advanced colorectal cancer appears as an everyday challenge for surgeons and oncologists. The new oncologic procedures invented last years led to significant therapeutic improvement. Combination of neoadjuvant chemotherapy with biological answer modifiers increased greatly the clinical response rate given to cytoreductive therapy. Due to these facts the complete disappearance of liver metastases can be observed more and more frequently after 2000. Literature of metastasis surgery clarified the exact difference between clinical and pathological response, at the same time exact criteria of the connections between two responses have been set. In complex treatment of colorectal liver metastases complete pathological response became the end point of treatment. AIMS AND METHODS: Between January, 2009 and August, 2010, 39 patients with originally non resectable liver metastases (colorectal cancer origin) have been studied. All patients were treated by neoadjuvant chemo- and targeted therapy before the resection of liver. RESULTS: Complete pathological response has been observed in 3 patients, major pathological response (necrosis: >50%) in 11 patients, minor pathological answer (necrosis <50%) in 22 patients and finally no necrosis at all in 3 patients. CONCLUSIONS: Complete pathological response can be regarded as the final goal of neoadjuvant targeted therapy. Pathological response seems to be the most important prognostic factor which reflects long-term survival after the R0 resection of liver metastases. Complete disappearance of liver metastases is an undesirable side effect of oncological therapy, which causes difficulties during surgical intervention. In this term the overtreatment of patients resulting in a disappearance of metastases should be avoided. Multidisciplinary team is responsible for the indication of resection of liver metastases in time before their disappearance.