West Nile virus surveillance in mosquitoes, April to October 2013, Vojvodina province, Serbia: implications for the 2014 season.

After the West Nile virus (WNV) outbreak in 2012, we collected mosquito samples from Vojvodina province, Serbia, in 2013. We found high WNV infection rate in two species, Culex pipiens and Anopheles maculipennis. Phylogenetic analysis showed that Serbian WNV strains from 2013 were most closely related to Italian and Greek strains isolated in 2012 and 2010, respectively. Public health authorities should be aware of a potentially increased risk of WNV activity during the 2014 season.

Novel European lineages of bat astroviruses identified in Hungary.

KEYWORDS: bat, astroviruses, coronaviruses, snRT-PCR, Hungary.

Comparison of immune activation of the COVID vaccines: ChAdOx1, BNT162b2, mRNA-1273, BBIBP-CorV, and Gam-COVID-Vac from serological human samples in Hungary showed higher protection after mRNA-based immunization.

OBJECTIVE: To gain insight into the different protective mechanisms of approved vaccines, this study focuses on the comparison of humoral and cellular immune responses of five widely used vaccines including ChAdOx1 (AZD1222, AstraZeneca), BNT162b2 (Pfizer), mRNA-1273 (Moderna), BBIBP-CorV (Sinopharm), and Gam-COVID-Vac (Sputnik V). MATERIALS AND METHODS: Isolated plasma from 95 volunteers' blood samples was used to measure anti-SARS-CoV-2 humoral and cellular immune responses. Positive controls were recovered patients from COVID-19 (unvaccinated). Specific quantification kits for anti-nucleocapsid IgG, anti-Spike protein IgG, neutralizing antibodies as well as specific SARS-CoV-2 antigens for T-cell activation were used and Spearman correlation and matrix analyses were performed to compare overall immune responses. RESULTS: Nucleocapsid antibodies were significantly higher for the BBIBP-CorV and convalescent group when compared to other vaccines. In contrast, subjects vaccinated with BNT162b2 and mRNA-1273 presented significantly higher anti-spike IgG. In fact, 9.1% of convalescent, 4.5% of Gam-COVID-Vac, 28.6% of ChAdOx1, and 12.5% of BBIBP-CorV volunteers did not generate anti-spike IgG. Similarly, a positive correlation was observed after the neutralization assay. T-cell activation studies showed that mRNA-based vaccines induced a T-cell driven immune response in all cases, while 55% of convalescents,  8% of BNT162b1,  12,5% of mRNA-1273, 9% of Gam-COVID-Vac,  57% of ChAdOx1,  and  56% of BBIBP-CorV subjects presented no cellular response. Further correlation matrix analyses indicated that anti-spike IgG and neutralizing antibodies production, and T-cell activation follow the same trend after immunization. CONCLUSIONS: RNA-based vaccines induced the most robust adaptive immune activation against SARS-CoV-2 by promoting a significantly higher T-cell response, anti-spike IgG and neutralization levels. Vector-based vaccines protected against the virus at a comparable level to convalescent patients.

The phylogenetic relationships among some Randia (Rubiaceae) taxa.

Phylogenetic relationships among some Randia (Rubiaceae, Gardenieae) taxa were estimated based on sequence variation in the nuclear ribosomal internal transcribed spacers (ITS) and rps 16 intron (cpDNA). During the investigation of rpsl6 intron of 9 studied Central American Randia species, two well supported subclades were separated. Analysis of ITS data of 16 Randia species shows 3 major clades. A group of mainly lowland, South American Randia species is moderate supported (75%). Species from Mexico form a strongly supported (97%) clade, but the Central American and Mexican Randia species are low supported (58%). However the last two groups are well supported together (95%). The molecular delimination is well in line with the size of leaves combined with the texture of exocarp.

Prevalence and molecular characterization of human group C rotaviruses in Hungary.

BACKGROUND: Group C rotaviruses are recognized enteric pathogens of humans and animals. Human group C rotaviruses have been associated with sporadic episodes and large outbreaks of gastroenteritis in children and adults but their epidemiology and ecology are still unexplored. OBJECTIVES: To collect epidemiological data on group C rotavirus infections among children with gastroenteritis in Hungary and perform molecular characterization on the identified strains. STUDY DESIGN: Fecal samples were collected during the 2003 surveillance in Baranya County, Hungary. The presence of group C rotavirus RNA was investigated by polyacrylamide gel electrophoresis and by reverse transcription-nested polymerase chain reaction for the VP6 gene. The identified strains were further characterized by sequencing and phylogenetic analysis of the VP7, VP6, VP4, and NSP4 genes. RESULTS: Three of 472 samples (0.6%) tested positive for group C rotavirus. Two samples were selected for molecular analysis. Strains BaC 6104/03 and BaC 11549/03 displayed an overall identity of >99.8% and 99.3% at the nucleotide and amino acid level, respectively. The VP7 of the strain BaC 6104/03 was most closely related (99.5% aa) to the Nigerian strain Jajeri, while the VP4s of strains BaC 6104/03 and BaC 11549/03 were more similar (98.1% aa) to strains Belem and 208, detected in Brazil and China, respectively. CONCLUSIONS: Based on this 1-year study, we conclude that group C rotaviruses are not of epidemiological relevance in the etiology of childhood acute gastroenteritis in Hungary. The low sequence divergence between the Hungarian strains suggested that a single group C rotavirus strain circulated in this period in the study area.

Pathologic assessment of response to chemotherapy in colorectal cancer liver metastases after hepatic resection: which method to use?

BACKGROUND: Patients with metastatic colorectal cancer receive chemotherapy prior liver resection more and more frequently. Histopathologic assessment methods of the resected specimen could evaluate the response to chemotherapy. In this study it is analyzed if these histopathologic changes are specific to preoperative chemotherapy and if these methods have correlation with survival. METHODS: Sixty three patients with available pathology slides, resected for colorectal cancer liver metastases were enrolled in this study. 46 patients (73%) received neoadjuvant chemotherapy. Five pathological evaluation methods were compared according to the literature: [1] residual tumor cell ratio, [2] tumor regression grade (TRG) scoring system, [3] modified tumor regression grade (mTRG) scoring system with the type of necrosis, [4] pattern of tumor regression and [5] the tumor thickness at the tumor-normal interface (TNI). RESULTS: Analyzing the pathological methods between the chemotherapy (CTX) and the non-chemotherapy group (NC), we found that that four evaluation methods showed significant and one showed strong correlation with the use of chemotherapy. (Residual tumor cell ratio: p = 0.08; TRG: p <0.01; mTRG: p = 0.03; pattern of tumor regression: p <0.01; TNI: p = 0.02). In the chemotherapy group none of the analyzed pathological methods showed significant correlation with progression free survival (PFS) or with overall survival (OS). Residual tumor cell ratio, TRG and the pattern of tumor cells showed positive but not significant correlation with OS and PFS and a slight difference in the group of patients with TNI <2 mm could be documented. CONCLUSIONS: Tumor regression grade (TRG) and tumor thickness at the tumor-normal interface (TNI) were the most useful methods for pathological response evaluation and these methods had some correlation with survival. According to these data, authors concluded, that a reproducible and well defined scoring system, based on different histopathological evaluation methods should be developed to predict more accurately the effect of neoadjuvant chemotherapy in CRCLM patients.

Mucosa of the Heister valve in cholelithiasis: transmission and scanning electron microscopic study.

Light, transmission, and scanning electron microscopic studies were performed on 24 gallbladders with thin walls, without inflammation, extirpated by cholecystectomy because of gallstones and on five, free from biliary diseases, obtained by autopsies. Examination of gallbladders and epithelium covering the valvula spiralis (Heister) of the ductus cysticus showed the surface structure of the epithelium covering the Heister valve to be mulberry-like and characteristically pleated, similar to the epithelial lining of the gallbladder. In cases of cholelithiasis, several denuded areas were detectable on the valves. The epithelial lining of the gallbladder was intact. As a new observation, we describe the appearance of worm-like processes on the lateral surface of detached epithelial cells, seen during the course of scanning electron microscopic study. These probably correspond to interdigitating cell junctions observed by transmission electron microscopy.

First clinical data of a natural immunomodulator in colorectal cancer.

BACKGROUND/AIMS: MSC (trade-name AVEMAR) is a per os applicable complex of multiple, biologically active molecules obtained from fermented wheat-germ extract. Preclinical studies suggest potent anti-metastatic activity and it has a favorable toxicity profile. It has been aimed in a pilot-scale, phase II clinical study to document whether or not MSC as a support to surgery or plus chemotherapy adds any therapeutic benefit compared to the same combination without MSC in colorectal cancer. METHODOLOGY: From 1998 to June 1999, 18 control patients and 12 consecutive colorectal cancer patients respectively, were enrolled into this study. All patients underwent curative surgery. The control group (18 patients) received no other therapy or adjuvant chemotherapy alone. The MSC group (12 patients) received MSC alone or plus adjuvant chemotherapy. Until now, the median follow-up has been 9 months. RESULTS: Interim data of the study document that in the MSC group no new metastases, neither hepatic nor other, have occurred, so far. On the contrary, several new metastases have developed in the control group. CONCLUSIONS: Orally administered MSC is a potent candidate to be regarded as a supportive therapy to surgery or plus chemotherapy for colorectal cancer patients.

The afferent circulation of the liver in patients with primary hepatocellular carcinoma.

BACKGROUND/AIMS: The data on the afferent circulation of the liver, in patients with primary hepatocellular carcinoma, are controversial or non-existent. MATERIALS AND METHODS: The authors measured hepatic arterial and portal venous flow intra-operatively by transit time ultrasonic volume flowmetry. RESULTS: In patients with primary hepatocellular carcinoma, the hepatic artery flow increased to 0.55 +/- 0.21 l, as compared with the control value of 0.37 +/- 102 l/min (p < 0.01). The portal venous flow decreased from 0.61 +/- 0.212 l/min to 0.47 +/- 0.203 l/min p < 0.01). Owing to the opposite changes in the afferent circulation, the total hepatic blood flow did not change significantly. The ratio of hepatic arterial flow to portal vein flow increased to 1.239 +/- 0.246 in patients with hepatocellular carcinoma, which is twice the basic control value (0.66 +/- 0.259 l/min). After resection, this ratio showed virtually no change. The surgical intervention, that is resection of the liver, did not significantly alter hepatic artery and portal venous flow, although total hepatic blood flow decreased significantly (p < 0.01). The pronounced increase in the ratio of hepatic arterial flow may be attributed to the decrease in portal venous flow caused by the primary hepatocellular carcinoma. The decrease in venous flow can most probably be explained by compression and infiltration of the intrahepatic branches of the portal vein. As we pointed out, the decrease in portal venous circulation leads to an increase in hepatic arterial flow. CONCLUSIONS: On the basis of our initial results, it seems probable that the ratio of the two circulations represents a diagnostic tool for the altered circulation in patients with hepatocellular carcinoma. The significance of this phenomena is not yet quite clear, but a review of the literature shows that similar observations have not been reported. In practical terms this phenomenon may be useful from the point of view of US and Doppler US diagnosis, e.g. in the case of a hypo-echoic or hyperechoir, mass in the liver, increased blood flow in the hepatic, artery, and decreased portal venous flow, a malignant liver tumor is virtually certain. The authors hypothesise that any pathology in the liver may lead to a primary decrease in PVF and a subsequent increase in HAF.

The interaction between hepatic arterial and portal venous blood flows; simultaneous measurement by transit time ultrasonic volume flowmetry.

Intra-operative measurement of the afferent circulation of the liver, namely hepatic artery and portal venous flow was carried out in 14 anesthetized patients with carcinoma of the splanchnic area, mainly in the head of the pancreas, by means of transit time ultrasonic volume flowmetry. The hepatic artery flow, portal venous flow and total hepatic flow were 0.377 +/- 0.10; 0.614 +/- 0.21; 0.992 +/- 0.2761/min, respectively. The ratio of hepatic arterial flow to portal venous flow was 0.66 +/- 0.259. There was a sharp, significant increase in hepatic arterial flow (29.8 +/- 6.1%, p < 0.01) after the temporary occlusion of portal vein, while the temporary occlusion of the hepatic artery did not have any significant effect on portal venous circulation. The interaction between hepatic arterial flow and portal venous flow is much in dispute, but, as the data presented here show, there is no doubt that the decrease in portal venous flow immediately gives rise to a significant increase in hepatic artery circulation.

Complications following major abdominal surgery in cirrhotic patients.

The morbidity and mortality associated with major abdominal surgical interventions in 34 histologically proven cirrhotic patients are analyzed by the authors. The surgical interventions were carried out as urgent, absolute and elective indications. Thirty-seven general and surgical complications were observed following major abdominal surgery in 34 cirrhotics. Seven out of 34 patients died, giving a mortality rate of 21%. Suture-line insufficiency, peritonitis, sepsis and other inflammatory processes turned out to be the most common complications. Statistical analysis showed that the Child criteria, prothrombin level and white blood cell count were useful prognostic factors.

The first histological demonstration of pancreatic oxidative stress in human acute pancreatitis.

Necrotizing acute pancreatitis is associated with an inflammatory explosion involving numerous pro-inflammatory mediator cascades and oxidative stress. Acinar oxygen free radical production aggravates pancreatic tissue damage, and promotes cellular adhesion molecule upregulation resulting in leukocyte adherence and activation. The cerium capture oxygen free radical histochemistry combined with reflectance confocal laser scanning microscopy allows the "in situ" histological demonstration of oxygen free radical formation in live tissues. Here we present a case report, where oxidative stress is demonstrated on a histological level for the first time in human acute pancreatitis. A 44-year-old male patient suffering from acute exacerbation of his chronic pancreatitis developed a pancreato-pleural fistula with amylase-rich left pleural exudate causing respiratory compromise. Subsequent to an urgent thoracic decompression a distal pancreatectomy and splenectomy was performed with the closure of abdomino-thoracic fistula. The postoperative course was uneventful, except for a transient pancreatico-cutaneous fistula, which healed after conservative treatment. To carry out cerium capture oxygen free radical histochemistry the resected pancreas specimen was readily perfused with cerium-chloride solution through the arteries on the resection surface. Frozen sections were cut, E-, P-selectin, ICAM and VCAM were labeled by immunofluorescence. The tumor-free margin of an identically treated pancreas carcinoma specimen served as a control. Intrapancreatic oxidative stress and cellular adhesion molecule expression were detected by confocal laser scanning microscopy. Numerous pancreatic acini and neighboring capillaries showed oxygen free radical-derived cerium-perhy-droxide depositions corresponding to strong local oxidative stress. Acinar cytoplasmic reflectance signals suggested xanthine-oxidase as a source of oxygen free radicals. These areas presented considerably increased endothelial P-selectin expression with adherent, oxygen free radical-producing polymorphonuclear leukocytes displaying pericellular cerium-reflectance. Modest ICAM upregulation was noted, E-selectin and VCAM expression was negligible. The control pancreas specimen showed minimal oxidative stress with weak, focal P-selectin expression. The development of deleterious pancreatic oxidative stress was based on indirect evidence in human acute pancreatitis. To the best of our knowledge this is the first report demonstrating persistent intrapancreatic oxidative stress histologically in human acute pancreatitis. We have noted P-selectin overexpression with a preponderance in the areas of acinar oxidative stress.

Cannulation of the cervical thoracic duct in the rat.

A method is described for cannulation of the cervical part of the thoracic duct in the rat. The volume or lymph flowing from the cervical cannula is approximately 80% higher and its protein concentration by about 18% lower than that of the lymph collected from a cannula introduced into the abdominal part of the thoracic duct.

Bile constituents in blood and lymph during biliary obstruction. I. The dynamics of absorption and transport of ions and colloid molecules.

Na125I and 131I-labeled albumin was infused in dogs into the common bile duct at pressures of 20 to 25 and 40 mmHg. At 40 mmHg, the amounts of the iodide ion and labeled albumin in circulating plasma were, after correction for the secondary loss from the circulation, nearly identical. At 20-25 mmHg more iodid than labeled albumin was found in the circulation. In thoracic duct lymph the same fraction of the infused amount of albumin was recovered at both pressures. Lymphatic concentrations of albumin were in both types of experiments substantially higher than plasma concentrations. It is concluded, that at increased pressure fluid leaks first from the small biliary ducts into the Mall's spaces. In consequence of water absorption and the diffusion of ions and small molecules into the blood capillaries the concentrations of protein or protein bound molecules in this part of the hepatic interstitial fluid increases. This is reflected in their high concentration in the lymph. If bile pressure rises further, fluid leaks also into the Disse's spaces. This leads to a bulk flow of solvent and solutes into the sinusoids and to the near disappearance of the differences in the venous transport of ions and colloids.

The effect of lymphostasis on bile flow and composition in the rat.

The ligation of the thoracic duct and of the hepatic lymph vessels leads in the rat to a decease of bile flow and bile salt output. Bilirublin excretion was temporarily reduced both in lymphostasis and in sham operated animals, but bilirubin concentration in the bile rose progressively and after 3 hours bilirubin output returned in both groups to the control level.

Bile constituents in blood and lymph during biliary obstruction. II. The absorption and transport of bile acids and bilirubin.

The lymphatic and venous transport of bilirubin and total bile acid was examined in dogs after the occlusion of the common bile duct. Lymphatic concentrations of both substances attained maximum levels between the 4 th and 6 th hours, but remained during the entire time of observation (24 hours) above plasma concentrations. The concentrations in blood plasma rose more slowly, but continuously. The amounts of both substances transported by the lymphatics rose steadly for 6 or 8 hours respectively and exceeded after 2 hours of occlusion the amounts transported by the veins. The results are explained by the changes in bilirubin and bile acid formation and secretion during biliary obstruction and on the basis of observations made in experiments with electrolyte and colloid infusions into the biliary passages.

Biliary decompression in the treatment of acute pancreatitis.

The etiology and treatment of acute pancreatitis are discussed. The two main principles are conservative and surgical treatment. Biliary decompression was chosen as an elective method in the surgical treatment of acute pancreatitis, if signs of bile stasis and tension of the common bile duct could be seen. Peritoneal or peripancreatic dialysis were also applied. Five patients were treated in this way; according to literature and our experience it is a good, short and curative intervention for the treatment of acute pancreatitis.

Genetic variation within and relationships among five subpopulations of Slovak Thoroughbred.

Genetic variation at six microsatellite loci was analysed for five Thoroughbred subpopulations to determine the magnitude of genetic differentiation and the genetic relationships among the subpopulations. Significant deviations from Hardy-Weinberg equilibrium were shown for a number of locus-population combinations, with all subpopulations. The genetic diversities and relationships of five Thoroughbred subpopulations were evaluated using six microsatellites recommended by the International Society of Animal Genetics (ISAG). The allele frequencies, the effective numbers of alleles, and the observed and expected heterozygosities were calculated. POPGENE v. 1.31 (Yeh et al., 1997) was used to test for deviations from the Hardy-Weinberg (H-W) equilibrium and to assign F(IS) estimates (Weir, 1990). The utility of microsatellites for evaluating genetic diversity of horses is discussed.

[Organizational difficulties in liver transplantation]. / A májtranszplantáció szervezési nehézségei.

Author overviews the evolution of liver transplantation from the point of view of organization in North America, in Europe and in Hungary partly on the basis of his own experience. It can be stated, that in North America the liver transplantation has become the universal method for the treatment of end-stage liver diseases, only after 20 year long period since the beginning. This first 20 years could be characterized by the monopoly of few giant transplant centers. This period was different and shorter in Europe due to integration run by Eurotransplant and Council of Europe. In Hungary the need and hope for a well organized, functioning liver transplant programme has become reality after difficult long period. Individual efforts, organizations, scientific-, clinical-, experimental work are being regarded valuable experience, because due to this effort some liver patients can live with good quality of life, the results are included to the present programme, on the other hand this accumulated knowledge can serve as a basis for further transplant activity.