Presence of Newcastle disease viruses of sub-genotypes Vc and VIn in backyard chickens and in apparently healthy wild birds from Mexico in 2017.

Virulent Newcastle disease viruses (NDV) have been present in Mexico since 1946, and recently, multiple outbreaks have been reported in the country. Here, we characterized eleven NDV isolated from apparently healthy wild birds and backyard chickens in three different locations of Jalisco, Mexico in 2017. Total RNA from NDV was reverse-transcribed, and 1285 nucleotides, which includes 3/4 of the fusion gene, was amplified and sequenced using a long-read MinION sequencing method. The sequences were 99.99-100% identical to the corresponding region obtained using the Illumina MiSeq. Phylogenetic analysis using MinION sequences demonstrated that nine virulent NDV from wild birds belonged to sub-genotypes Vc and VIn, and two backyard chicken isolates were of sub-genotype Vc. The sub-genotype Vc viruses had nucleotide sequence identity that ranged from 97.7 to 98% to a virus of the same sub-genotype isolated from a chicken in Mexico in 2010. Three viruses from pigeons had 96.3-98.7% nucleotide identity to sub-genotype VIn pigeon viruses, commonly referred to as pigeon paramyxovirus, isolated in the USA during 2000-2016. This study demonstrates that viruses of sub-genotype Vc are still present in Mexico, and the detection of this sub-genotype in both chickens and wild birds suggests that transmission among these species may represent a biosecurity risk. This is the first detection and complete genome sequencing of genotype VI NDV from Mexico. In addition, the utilization of an optimized long-read sequencing method for rapid virulence and genotype identification using the Oxford nanopore MinION system is demonstrated.

An Epizootiological Report of the Re-emergence and Spread of a Lineage of Virulent Newcastle Disease Virus into Eastern Europe.

A number of contemporary outbreaks of Newcastle disease (ND) in Israel, Turkey, Georgia and Bulgaria have all been caused by a very similar viruses related to lineage 5a (genotype VIIa). Comparison with published ND virus (NDV) sequences suggests that this virus strain originated in South-East Asia and on introduction has circulated widely in backyard poultry in the Middle East and into Eastern Europe. An intracerebral pathogenicity index of 1.9 was obtained for a representative isolate from Bulgaria. In addition, the International Reference Laboratory for ND has characterized a molecular epidemiologically linked virus that has been reported to have caused disease in well-vaccinated broiler chickens in Pakistan. In the 1990s, another strain from the 5a lineage NDV was introduced into Europe and spread across the continent causing numerous outbreaks up to 1999. Despite improved controls, including good diagnostic tests and widespread vaccination, in commercial poultry, the novel circulating NDV strains described here have been established widely in the region and represent an increased risk for similar disease outbreak events to reoccur within the EU.

Valorization of cruor slaughterhouse by-product by enzymatic hydrolysis for the production of antibacterial peptides: focus on α 1-32 family peptides mechanism and kinetics modeling.

Bovine hemoglobin is the major component of the cruor (slaughterhouse by-product) and can be considered as an important source of active peptides that could be obtained by pepsic hydrolysis. The kinetics of appearance and disappearance of several antibacterial peptides from α 1-32 family during hydrolysis of synthesized α 1-32 peptide, of purified bovine hemoglobin and of cruor was studied, and reaction scheme for the hydrolysis of α 1-32 family peptides from these three sources was determined. On this basis, a mathematical model was proposed to predict the concentration of each peptide of interest of this family depending on hydrolysis time, and also on temperature (in the range 15-37 °C), pH (in the range 3.5-5.5) and enzyme to substrate ratio (in the range 1/50-1/200 for the synthesized peptide and 1/5-1/20 for purified bovine hemoglobin and cruor). Apparent rate constants of reactions were determined by applying the model on a set of experimental data and it was shown that they depended on the temperature according to Arrhenius's law, that their dependence on the pH was linear, and that enzyme to substrate ratio influence was limited (in the studied range).

Mechanism and kinetics modeling of the enzymatic hydrolysis of α1-32 antibacterial peptide.

Several antibacterial peptides can be obtained by enzymatic hydrolysis of the α chain of bovine hemoglobin. The kinetics of α1-32 peptide hydrolysis by pepsin was studied at several temperatures (15, 23 and 37 °C). Intermediate and final peptides were identified, and their antibacterial activity was assessed against four bacterial species. Evolution of generated peptides concentration enabled to propose a reaction pathway describing the parallel and consecutive reactions taking place during the hydrolysis. A mathematical model, based on the proposed mechanism, was developed to describe the kinetics of generated peptides during α1-32 hydrolysis. The constants of the main reactions were identified based on the experimental data, and their dependence on temperature was established using Arrhenius-type equations. Validation of the proposed model was performed by predicting kinetics of α1-32 peptide hydrolysis at 30 °C (all other experimental conditions being unchanged) with a good accuracy. This mathematical model could allow defining the optimal conditions for the production of various intermediate peptides with antibacterial activity from peptic hydrolysis of α1-32 peptide.

Diagnosis of liver hemangioma.

BACKGROUND: Benign liver tumors are now being diagnosed frequently with the advent of the greater use of imaging investigations. The most common lesion is the liver hemangioma 0.4-7.3%, with an incidence rate at autopsy ranging from 3% to 20%. MATERIAL AND METHODS: One hundred and one patients underwent operative treatment in the Department of General and Liver-pancreatic Surgery, UH "Alexandrovska" Sofia from 1995 to April 2013. There were 74 (73.3%) females and 27 (26.7%) males. Mean age was 50.7 years (range 25 to 77). Seventy-four patients (73.3%) had a solitary tumor. Methods for diagnosis included history of the disease, routine haematological and liver function tests, ultrasonography, CT, MRI or a combination of more than one technique. RESULTS: The diagnostic sensitivity of the imaging procedures was U/S 96.9%, CT scanning 98.3% and MRI 100% .The diagnostic specificity--U/S 60.3%, CT scanning 55.0%, MRI 85.7%. CONCLUSION: Hepatic hemangioma is diagnosed in most patients using non-invasive studies such as US, CT and MRI or a combination of them.

Neurocutaneous metacarpal flaps.

The article describes the possibility of covering large skin defects of the fingers by the use of fasciocutaneous flap from the back of the hand. It is pedicled on a sensory nerve branch and its paraneural tissue. The flap relies on blood supply from the constant para- and intraneural vascular network which gives perforator branches towards the overlying skin as well as on the constant anastomoses between the dorsal and volar arterial pool. We present a patient with a skin defect along the whole radial side of the index who underwent such reconstruction.

Perforator-based forearm island flap.

Reconstruction of soft tissue defects of the hand with exposed tendons, nerves, joints and bones could be quite a challenge for the surgeon. Only adequate coverage can save their functions. In our case we present a patient with adduction contracture of the right thumb after II degree burns, treated with perforator-based forearm flap. This flap relies only on perforating branches (5 to 11 in number) arising from the radial artery and located proximal to the radial styloid (2 to 7 cm). The main advantages of this method are that it is a single-stage operative technique, it is applicable in emergency conditions, and the main artery is spared. The cosmetic effect of the donor site could be considered a disadvantage of this approach.

Climatic adaptation of Drosophila buzzatii populations in southeast Australia.

Variation in 19 traits possibly relevant for thermal adaptation was studied in 11 populations of Drosophila buzzatii collected in southeast Australia. Using stepwise multiple regression, the variation was compared to variation in geographic coordinates and to a set of climatic variables estimated for each collection site. For 13 of the traits, a significant part of the variation was explained by climatic variables and/or geographic coordinates, suggesting directional selection for adaptation to the environment in the majority of traits studied. In 10 of the traits, both geographic coordinates and climatic variables explained significant proportions of the variation, with R2 ranging from 0.075 to 0.58. Although larvae, pupae and adults of D. buzzatii share a common habitat, the measured traits were not correlated across life stages and gender. Also, there seemed to be special conditions in marginal populations near species borders, giving rise to nonlinear relations with latitude. Climate apparently does influence the adaptive evolution of the traits studied, but they also are affected by other factors that vary with latitude, longitude and distance to coast. These results highlight the complex challenges imposed by the environment on the adaptive process.

[Postoperative entero-cutaneus fistulas--a study of 24 cases].

AIM: To perform a retrospective study on the experience of the Clinic of General, Liver, Biliary and Pancreatic Surgery, Alexandrovska Hospital in the diagnosis and treatment of patients with acquired entero-cutaneus fistulas and to analyse the early postoperative results. MATERIAL AND METHODS: We treated 24 patients with postoperative entero-cutaneous fistulas in the period Jan., 2000 - Aug., 2004. The male sex and the age above 50 were dominent. The primary disease was of oncological origin in 22 cases (91.66%). Anemia, hypoprotenemia and hypoalbuminemia were predisposing and risk factors in 19 cases (79.17%). Fistulography was routinely used for establishing the diagnosis. Surgical treatment was received by all the 6 patients (100%) with small bowel fistulas and by 15 from the 18 patients with large bowel fistulas (83.33%). EARLY POSTOPERATIVE RESULTS: The mortality rate was 4.54% (1 fatal outcome from 22 operated patients). The morbidity rate was 36,36% (8 cases) but none of the complications needed a reoperation. CONCLUSIONS: 1. The most frequent causal factor for entero-cutaneus fistulas in our material was an anastomotic leakige after bowel resection (95.83%); 2. The risk for such a coplication is higher in cancer patients with other predisposing pathological factors; 3. From the pahtological and clinical point of view the entero-cutaneus fistulas caused significant disturbances in base-alkaline and electrolyte balance, malnutrition and cahexia; 4. The clinical signs and the fistulography prooved to be reliable diagnostic methods; 5. The preoperative substitution and nutritional therapy had a significant benefit on the postoperative results, especially in cases of small bowel fistulas; 6. The open surgery was the main therapeutical method in the complex treatment of those patients.

[Acquierd entero-cutaneous fistulas--diagnostic and therapeutic approach].

The acquired entero-cutaneous fistulas are a current problem in the field of abdominal surgery. Most of them are postoperative--after an intestinal resection and/or anastomosis. Crohn's disease and coplicated colonic diverticulosis rank second as causal factors. The risk factors for the development of an entero-cutaneous fistula and for the poor prognosis at the same time are the next: (1) Most of the patients suffer of severe main and co-exhisting diseases; (2) The presence of previous laparotomies, radion and chemotherapy, significant disturbances in the base-acid and water-electrolyte balance; (3) The fistula itself worsens these disturbances and may doom to fail the substitutional and nutritional therapy. The diagnostics of a fistula, including its location is comparatively easy. The surgical treatment plays an important role in the therapeutic scheme, especially in cases of high-output fistulas of the small intestin. At the same time the adequate total parenteral nutrition and correction of the base-acid and water-electrolyte disbalance is also very important. Recently there are some new diagnostic methods and alternatives of the basic surgical procedures, some of them quite contraversial.

[Anterior rectal resection--early postoperative results]. / Predna rezektsiia na rektuma--ranni sledoperativni rezultati.

BACKGROUND: The development of the mechanic staplers gave the opportunity to perform sphincter-saving operations in patients with middle and low rectal carcinoma. It has been gained experience in performing this operation as the anastomosis was sewed in both ways--manually and mechanically. PURPOSE: The purpose of this retrospective research is to analyse the early postoperative results after anterior rectal resection with manually and mechanically made anastomosis. MATERIAL AND METHODS: For the period 01.01.1995-01.012000 in the DEPARTMENT OF GENERAL, LIVER AND PANCREATIC SURGERY, 69 patients with rectal carcinoma underwent the operation anterior rectal resection. In 60 cases (89.96%) the anastomosis was made manually and in 9 cases (13.04)--with mechanical staplers. Both patient groups are studied and classified by sex, age, tumor localisation, type of anastomosis, early postoperative results. RESULTS: From the group with manually sewn anastomosis 6/10%/ of the patients had complications in the early postoperative period, one of them terminated (1.7%). From the second group 3 (33.3%) of the patients had complications in the early postoperative period. The average postoperative hospitalisation was 13.5 vs. 19.2 days. CONCLUSION: The results achieved in anterior rectal resection depend on a complex of factors among which more important is the early diagnosis of the disease and the skills of the surgeon in coloproctology.

Stage-specific activity of the Leishmania major CRK3 kinase and functional rescue of a Schizosaccharomyces pombe cdc2 mutant.

Cell cycle control by cdc2-related kinases (CRKs) is essential to the regulation of cell proliferation and developmental processes in many organisms. Alternating phases of growth, arrest, and differentiation are characteristics of the infectious cycle of many trypanosomatid parasites, raising the possibility that members of the trypanosomatid CRK gene family participate in the regulation of these essential processes. Here we describe properties of the CRK3 gene from Leishmania major, which encodes a 36 kDa protein kinase showing 60% amino acid sequence identity with human CDK2, including several conserved sites implicated in regulation of kinase activity. CRK3 mRNA was constitutively expressed throughout the parasite life cycle, but histone H1 kinase activity of an epitope tagged CRK3 protein was greater in log-phase than in stationary-phase promastigotes. When integrated into the genome and expressed at the optimal level, CRK3 was able to rescue the growth defect of a Schizosaccharomyces pombe cdc2 mutant (cdc2-33(ts)), indicating that CRK3 is a functional homolog of cdc2. Mutants of CRK3 at several key regulatory residues showed the expected dominant negative effects on the S. pombe mutant. This is the first example of functional expression of a trypanosomatid CRK in yeast, opening the way for further genetic studies within this amenable organism.

The role of fnx1, a fission yeast multidrug resistance protein, in the transition of cells to a quiescent G0 state.

Most microorganisms live in conditions of nutrient limitation in their natural habitats. When exposed to these conditions they respond with physiological and morphological changes that enable them to survive. To obtain insights into the molecular mechanisms of this response a systematic genetic screen was performed to identify genes that when overexpressed can induce a starvation-like response in the yeast species Schizosaccharomyces pombe. One gene that meets these criteria, fnx1(+), induces, transcriptionally correlates with, and is required for the entry into the quiescent G0 state that is normally induced by nitrogen starvation. fnx1(+) encodes a protein with sequence similarity to the proton-driven plasma membrane transporters from the multidrug resistance group of the major facilitator superfamily of proteins. We propose that fnx1(+) plays a role in the entry into G0, possibly by facilitating the release of a signaling substance into the environment as a means of cell-to-cell communication.

Perturbations in the spi1p GTPase cycle of Schizosaccharomyces pombe through its GTPase-activating protein and guanine nucleotide exchange factor components result in similar phenotypic consequences.

spi1p of Schizosaccharomyces pombe is a structural homolog of the mammalian GTPase Ran. The distribution between the GTP- and GDP-bound forms of the protein is regulated by evolutionarily conserved gene products, rna1p and pim1p, functioning as GTPase-activating protein (GAP) and guanine nucleotide exchange factor (GEF), respectively. Antibodies to spi1p, pim1p, and rna1p were generated and used to demonstrate that pim1p is exclusively nuclear, while rna1p is cytoplasmic. A loss of pim1p GEF activity or an increase in the rna1p GAP activity correlates with a change in the localization of the GTPase from predominantly nuclear to uniformly distributed, suggesting that the two forms are topologically segregated and that the nucleotide-bound state of spi1p may dictate its intracellular localization. We demonstrate that the phenotype of cells overproducing the GAP resembles the previously reported phenotype of mutants with alterations in the GEF: the cells are arrested in the cell cycle as septated, binucleated cells with highly condensed chromatin, fragmented nuclear envelopes, and abnormally wide septa. Consistent with the expectation that either an increased dosage of the GAP or a mutation in the GEF would lead to an increase of the spi1p-GDP/spi1p-GTP ratio relative to that of wild-type cells, overexpression of the GAP together with a mutation in the GEF is synthetically lethal. The similar phenotypic consequences of altering the functioning of the nuclear GEF or the cytoplasmic GAP suggest that there is a single pool of the spi1p GTPase that shuttles between the nucleus and the cytoplasm. Phenotypically, rna1 null mutants, in which spi1p-GTP would be expected to accumulate, resemble pim1(ts) and rna1p-overproducing cells, in which spi1p-GDP would be expected to accumulate. Taken together, these results support the hypothesis that the balance between the GDP- and GTP-bound forms of spi1p mediates the host of nuclear processes that are adversely affected when the functioning of different components of this system is perturbed in various organisms.

Comparison of the fatty acids of the tunicate Botryllus schlosseri from the Black Sea with two associated bacterial strains.

The fatty acid composition of the tunicate Botryllus schlosseri and of two bacterial strains found within the tunicate, namely Vibrio parahaemolyticus and of an associated but previously unreported gram positive cocci were studied. The polyunsaturated fatty acids 6,9,12-octadecatrienoic acid, 5,8,11,14,17-eicosapentaenoic acid, and 4,7,10,13,16,19-docosahexaenoic acid were particularly abundant in B. schlosseri and were not detected in the two bacterial strains found in the tunicate. The iso/anteiso pair, 13-methyltetradecanoic acid and 12-methyltetradecanoic acid, were the principal fatty acids in the gram positive cocci, and the 9- and 11-hexadecenoic acids were particularly abundant in V. parahaemolyticus. The diunsaturated fatty acid 9,12-octadecadienoic acid was also shown to be present in V. parahaemolyticus. The fatty acid composition of a third bacterial strain, characterized as either a Pseudomonas or an Alteromonas species, and shown to be present only in the sea water from the Black Sea and not in B. schlosseri, is also reported. This is the first investigation on fatty acids from Black Sea bacteria.

[Chlamydia infection in Reiter's syndrome]. / Untersuchungen über die Chlamydieninfektion beim Reitersyndrom (RS).

We report on 129 patients (109 females, 21 males) clinically suspect of Reiter's syndrome (RS).52 (48%) of the patients were positive for C. trachomatis. In 39.8% of the patients, we found antibodies against chlamydial complement with the titers 1:4 to 1:16. The titers 1:16 and higher were present in 33.3% of the men and in 22.2% of the women. Direct electron microscopy of urethral biopsies and synovial fluids revealed chlamydial elementary and initial bodies in 54.1% of the cases. Cytological investigations showed chlamydial inclusions in 38% of the patients. One strain of C. trachomatis could be isolated from urethral samples of two men in chicken embryos. HLA-B27 antigen was found in 86% of the patients positive for Chlamydia and and in 50% of those negative for Chlamydia.

[Associated vaccination of pigs against swine fever, erysipelas and Aujeszky's disease]. / Asotsiirano vaksinirane na praseta sreshtu chuma, cervenka i bolestta na Aueski.

Tested was the simultaneous vaccination of pigs against swine fever, swine erysipelas, and Aujeszky's disease. Parallel to this experiment a second group of test pigs were treated with each of the three vaccines at twenty-day intervals. The effect of vaccination was assessed through clinical observations with the test groups and by challenging part of the animals with virulent strains of the swine fever and erysipelas agents. It was found that the successive application of the three vaccines at 20-day intervals (at the age of 50, 70, and 90 days) could induce immunity that lasted for a longer period than the immunity resulting from the simultaneous treatment at the age of 90 days. In order to protect the pigs for sure up to the end of the fattening period a revaccination against swine fever with the simultaneous use of the vaccine against swine erysipelas in both cases was necessary.

[Commercial method of aerogenic immunization of pigs against swine fever]. / Promishlen metod ze aerogenno imunizirane na praseta protiv chuma.

An industrial method is presented through a model with a chamber for aerogenic immunization of about 250 pigs, aged 2.5 months. The experiments were carried out with more than 1,000 pigs of the Camborough hybrid under industrial conditions of raising. The process of immunization was feasibly carried out when the pigs passed through the chamber using aerosol at mixing them upon transference from the growing section to the fattening one, according to the provisions of the raising technology. A lapinized K-vaccine against fever, obtained from rabbits or tissue cultures was used in a dispersed state. Essential proved the high titer of the vaccine. Used were the aerosol devices DAG-2 or Microjet Fogmaster mounted at 1.5 m above the chamber floor. They guarantee the production of highly dispersed aerosol of an average arithmetic size of 7-10 microns, its concentration reaching 3,000,000 particles per 1 cm3. The animals were exposed for 30 min with Fogmaster and 45 min with DAG-2. It was shown that they tolerated perfectly this group immunization. A single procedure with the vaccine conferred immunity at 2.10(5) LT50 and protected the pigs from fever over the entire fattening period. The method was evaluated as a biotechnology.

[Clinical etiological findings in chlamydial urethritis in men]. / Klinisch ätiologische Befunde bei Chlamydienurethritiden bei Männern.

120 patients with abacterial urethritis were studied for chlamydia. Clinical, cytological, serologic and electronenmicroscopic tests were performed. 66.60% of the tested persons were seropositive for C. trachomatis, 32.50% of all patients and 48.80% of the seropositive patients showed positive cytological tests. Direct electron microscopy of biopsy material of the urethra of 30 patients revealed chlamydial bodies in high concentrations. The incubation period was about 1-2 weeks. Oxytetracycline was used for 8 days, but had to be one- to four times repeated in resistent cases.