Research Note: An overview on distribution of fowl adenoviruses.

Fowl adenoviruses (FAdV), detected during routine diagnostic investigations from 38 countries (5 continents) over a decade, were partially sequenced and grouped by phylogenetic analysis. The partial polymerase gene nucleotide sequences of the 365 fowl adenovirus isolates resulted in the following species distribution: 11% FAdV-A; 3% FAdV-B; 2% FAdV-C; 34% FAdV-D; and 50% FAdV-E. Noticeably, only 79 of the detected strains could be associated with adenovirus-specific pathologic conditions: 62 (79%) with inclusion body hepatitis; 9 (11%) with gizzard erosion; and 8 (10%) with hepatitis hydropericardium syndrome. The remainder of the FAdV strains was detected as concomitant infection from other disease conditions almost exclusively in boilers of 27 to 42 d of age: the majority of them was FAdV-E followed by FAdV-D, and to a lesser extent of FAdV-A, B, and C, the latter ones have not been associated with any of the established adenovirus-caused syndromes in our collection. The highest ratio of coinfections was observed for FAdV-B (62%), while it was about 30% for the rest of the FAdV species. The most frequent coinfection, in connection with all FAdV species, was with the avian infectious bronchitis virus. The presented database will serve as the basis for comparative whole genome and cross-neutralization analysis of selected FAdV isolates.

Advancement in vaccination against Newcastle disease: recombinant HVT NDV provides high clinical protection and reduces challenge virus shedding with the absence of vaccine reactions.

Newcastle disease (ND) is a highly contagious disease of chickens causing significant economic losses worldwide. Due to the limitation in their efficacy, current vaccination strategies against ND need improvements. This study aimed to evaluate a new-generation ND vaccine for its efficacy in providing clinical protection and reducing virus shedding after challenge. Broiler chickens were vaccinated in ovo or subcutaneously at hatch with a turkey herpesvirus-based recombinant vaccine (rHVT) expressing a key protective antigen (F glycoprotein) of Newcastle disease virus (NDV). Groups of birds were challenged at 20, 27, and 40 days of age with a genotype V viscerotropic velogenic NDV strain. Protection was 57% and 81%, 100% and 95%, and 100% and 100% after the subsequent challenges in the in ovo and subcutaneously vaccinated chickens, respectively. Humoral immune response to vaccination could be detected from 3-4 wk of age. Challenge virus shedding was lower and gradually decreased over time in the vaccinated birds compared to the unvaccinated control chickens. In spite of the phylogenetic distance between the NDV F gene inserted into the vector vaccine and the challenge virus (genotype I and V, respectively), the rHVT NDV vaccine provided good clinical protection and significantly reduced challenge virus shedding.

Bioplastic production using wood mill effluents as feedstock.

Fibreboard production is one of the most important industrial activities in Galicia (Spain). Great amounts of wastewater are generated, with properties depending on the type of wood, treatment process, final product and water reusing, among others. These effluents are characterized by a high chemical oxygen demand, low pH and nutrients limitation. Although anaerobic digestion is one of the most suitable processes for the treatment, lately bioplastics production (mainly polyhydroxyalkanoates) from wastewaters with mixed cultures is being evaluated. Substrate requirements for these processes consist of high organic matter content and low nutrient concentration. Therefore, wood mill effluents could be a suitable feedstock. In this work, the possibility of producing bioplastics from to wood mill effluents is evaluated. First, wood mill effluent was converted to volatile fatty acids in an acidogenic reactor operated at two different hydraulic retention times of 1 and 1.5 d. The acidification percentage obtained was 37% and 42%, respectively. Then, aerobic batch assays were performed using fermented wood mill effluents obtained at different hydraulic retention times. Assays were developed using different cultures as inoculums. The maximum storage yield of 0.57 Cmmol/Cmmol was obtained when when the culture was enriched on a synthetic media.

Valuable product production from wood mill effluents.

Fibreboard production is one of the most important industrial activities in Galicia (Spain). Great amounts of wastewater are generated, with properties depending on the type of wood, treatment process, final product and water reusing, among others. These effluents are characterized by a high chemical oxygen demand (COD), low pH and nutrients limitation. Aerobic and anaerobic processes have been used for their treatment. Presently, bioplastics production (mainly polyhydroxyalkanoates or PHA) from wastewaters with mixed cultures is being studied. Substrate requirements for these processes are a high organic matter content and low nutrient concentration. Therefore, wood mill effluents could be a suitable feedstock. PHA production from wastewaters is carried out in three steps. First, complex organic matter is converted into volatile fatty acids (VFA) through acidogenic fermentation. Then, VFA are used as substrate in an aerobic sequencing batch reactor (SBR), in which the enrichement of PHA producing bacteria from a mixed culture is favoured. Finally, the sludge from the SBR is fed with a pulse containing high VFA concentrations, resulting in PHA accumulation inside the cells. In this work, the possibility of applying this process to wood mill effluents is proposed. An acidification percentage of 37% and a storage yield (Y(STO)) of 0.23 Cmmol/Cmmol were obtained.

Comparative histopathology and immunohistochemistry of QX-like, Massachusetts and 793/B serotypes of infectious bronchitis virus infection in chickens.

The aim of this study was to compare experimentally the pathogenicity and tissue distribution of the recently emerged QX-like strain of infectious bronchitis virus (IBV) with the widespread M41 and 793/B serotypes of the virus. Histopathological and immunohistochemical methods were employed to define the main sites of virus replication. One-day-old specific pathogen free chickens were inoculated with five different QX-like strains, or with the M41 and 793/B IBV strains and monitored for 42 days post-infection. Tracheal lesions developed in all infected birds, confirming the ability of all of the tested strains to induce respiratory disease. Replication of the isolates in the alimentary tract was detected, but the infection did not cause significant gut lesions. Four of the five QX-like IBV strains induced severe kidney lesions. Dilation of the oviduct with accumulation of serum-like fluid in the lumen of this structure, reported previously from field cases of QX-like IBV infection, was observed following experimental infection with all of the five QX-like strains. Microscopical and immunohistochemical examination of the affected oviducts did not help to elucidate the pathogenesis of this lesion.

Bedside red cell volumetry by low-dose carboxyhaemoglobin dilution using expiratory gas analysis.

BACKGROUND: We developed a non-invasive, continuous, high-resolution method of measuring carboxyhaemoglobin fraction (COHb%) using expiratory gas analysis (EGA). We assessed whether application of EGA to carboxyhaemoglobin dilution provides red cell volume (RCV) measurement with accuracy equivalent to that of CO-haemoximetry, with a smaller infusion volume of carbon-monoxide-saturated autologous blood (COB). Method. We assessed the agreement between repeated COHb% measurements by EGA and simultaneous measurement by CO-haemoximetry, using Bland and Altman plot, in healthy subjects and patients with artificially controlled ventilation and no radiological evidence of pulmonary oedema or atelectasis. We assessed the agreement between RCV measurements by EGA with infusion of 20 ml of COB (RCVEGA) and RCV measurements by CO-haemoximetry with infusion of 100 ml of COB (RCVHEM), in healthy subjects. RESULTS: The 'limits of agreement' between COHb% measurement by EGA (1 min average) and CO-haemoximetry were -0.09 and 0.08% in healthy subjects, and -0.11 and 0.09% in patients. Given the resolution of CO-haemoximetry (0.1%), the accuracy of EGA was equivalent to or greater than that of CO-haemoximetry. The 'limits of agreement' between RCVEGA and RCVHEM were -0.14 and 0.15 litre. Given the average resolution of RCVHEM (0.14 litre), the accuracy of RCVEGA was equivalent to that of RCVHEM. CONCLUSION: EGA provided non-invasive, accurate, continuous, high-resolution COHb% measurements. Applying EGA to carboxyhaemoglobin dilution, we achieved RCV measurements with accuracy equivalent to that of CO-haemoximetry, with one-fifth of the COB infusion volume. However, clinical application of the method is limited to patients with no radiological evidence of pulmonary oedema or atelectasis.

Comparative pathological studies on domestic geese (Anser anser domestica) and Muscovy ducks (Cairina moschata) experimentally infected with parvovirus strains of goose and Muscovy duck origin.

Parvovirus infection of Muscovy ducks caused by a genetically and antigenically distinct virus has been reported from Germany, France, Israel, Hungary, some Asian countries and the USA. The pathological changes include those of degenerative skeletal muscle myopathy and myocarditis, hepatitis, sciatic neuritis and polioencephalomyelitis. In the study presented here, day-old and 3-week-old goslings and Muscovy ducks were infected experimentally with three different parvovirus strains (isolates of D-216/4 from the classical form of Derzsy's disease, D-190/3 from the enteric form of Derzsy's disease, and strain FM from the parvovirus disease of Muscovy ducks). All three parvovirus strains caused severe disease in both day-old and 3-week-old Muscovy ducks but in the goslings only the two strains of goose origin (D-216/4 and D-190/3) caused disease with high (90-100%) mortality when infection was performed at day old. Strain FM (of Muscovy duck origin) did not cause any clinical signs or pathological lesions in the goslings. In the day-old goslings and Muscovy ducks the principal pathological lesions were severe enteritis with necrosis of the epithelial cells (enterocytes) of the mucous membrane and the crypts of Lieberkühn, and the formation of intranuclear inclusion bodies. Other prominent lesions included hepatitis and atrophy (lymphocyte depletion) of the lymphoid organs (bursa of Fabricius, thymus, spleen). In goslings infected with the strain originating from the classical form of Derzsy's disease mild myocarditis was also detected. After infection at three weeks of age, growth retardation, feathering disorders, myocardial lesions (degeneration of cardiac muscle cells, lympho-histiocytic infiltration) and hepatitis were the most prominent lesions in both geese and Muscovy ducks. In addition to the lesions observed in the geese, muscle fibre degeneration, mild sciatic neuritis and polioencephalomyelitis were also observed in the Muscovy ducks infected with any of the three parvovirus strains.

Antigenic and genetic diversity of early European isolates of Infectious bursal disease virus prior to the emergence of the very virulent viruses: early European epidemiology of Infectious bursal disease virus revisited?

Eleven Polish and Hungarian isolates of Infectious bursal disease virus (IBDVs) obtained in the 70/80s (early IBDV) and in the 90s (recent IBDV) were characterized in an Antigen-Capture-ELISA with a panel of neutralizing monoclonal antibodies (Mabs), and by nucleotide sequencing of the VP2 variable domain (vVP2). The viruses were compared with reference IBDV strains, among others with Faragher 52/70 (F52/70, classical, isolated 1970), 89163 (typical very virulent-vvIBDV, isolated 1989) and 91168 (antigenically modified vvIBDV, isolated 1991). Only one of the early isolates (Hungarian strain P1) proved antigenically and genetically similar to F52/70. Other early isolates exhibited no reactivity versus Mabs 3, 4, 5 and/or 8 and had a common previously unrecognized combination of amino acid changes in vVP2. The recent isolates all proved antigenically and genetically related to typical vvIBDV strain 89163, except the Polish isolate 93/35 which proved related to the 91168 strain although no epidemiological relationship had been documented between these viruses in the field. Phylogenetic analysis confirmed that the non-P1 early IBDVs represent a previously unrecognized group among serotype 1 IBDVs. It is discussed whether these early isolates are derivatives of the F52/70-like viruses that might still be present in the field, or whether they represent early IBDV strains that might have been present prior to and progressively replaced by the F52/70-like viruses, as the latter have been replaced by vvIBDVs in the late eighties.

Guillain-Barré syndrome following herpes zoster in a patient with systemic sclerosis.

Abstract We report the case of a patient with systemic sclerosis (SSc) who developed Guillain-Barré syndrome (GBS) 6 weeks after herpes zoster. Muscle weakness developed first, and thereafter severely in the muscles in the same segment as the zoster. Serum anti-GM1 and -GD1b IgM autoantibodies were detected in the acute phase. The clinical course and the findings of nerve conduction studies and a sural nerve biopsy were compatible with GBS accompanied by underlying chronic polyneuropathy. SSc might have affected the neurological manifestation via the development of underlying neuropathy and a possible contribution to the autoimmune basis in GBS.

Regional difference of lipid distribution in brain of apolipoprotein E deficient mice.

According to recent knowledge, apolipoprotein E (apo E) plays a significant role in the homeostasis of intracellular cholesterol level in various tissues. Apo E deficient mice develop hyperlipidemia, and suffer from atherosclerosis in extracerebral blood vessels and neurodegeneration in the central nervous system. Furthermore, Walker et al. (Am. J. Path., 1997;151:1371-1377) demonstrated cerebral xanthomas of various sizes in the brain of apo E deficient mice. In the present study, it is illustrated that in the homozygous apo E deficient mice of advancing age, a great number of foamy macrophages extravasate from microvessels in thalamus and fimbria hippocampi, and scatter in the perivascular regions and migrate toward the ependyma, fimbria hippocampi, hippocampus, and thalamus. Here, it must be pointed out that under hyperlipidemia, although foamy macrophages made clusters in the perivascular region, the cerebral microvessels did not develop atherosclerosis. On the other hand, in the other cerebral regions such as cerebral cortex, caudoputamen, globus pallidus, and substantia nigra, macrophages did not appear and microvessels retained normal shapes, but the fluorescent granular perithelial (in short, FGP) cells accompanied by these vessels contained a certain amount of lipids. That is, in the cerebral cortex and caudoputamen, lipid components are detected in FGP cells and microglia, while in the globus pallidus and substantia nigra, they are mainly localized in astrocytes. The reason why the astrocytes in such defined regions contain, specifically, a high quantity of lipid components remains unsettled. Axonal degenerations are often represented in thalamus, globus pallidus, and substantia nigra. On the other hand, in the specimens of Wild-type mice, lipid components were observed only in FGP cells, and the vascular architecture took a normal profile. Any lipid laden macrophages and the axonal degenerations could not be detected through the cerebral parenchyma. Furthermore, it is also a noticeable finding that immunohistochemically, the FGP cells express a positive reaction against the antibody of apo E in the Wild-type mice, but those of homozygous apo E deficient mice are immunonegative. FGP cells are not only provided with the scavenger receptor, but also contribute to the lipid metabolism in the brain.

Correlation of clonal T cell expansion with disease activity in systemic lupus erythematosus.

It has been proposed that T cell activation may play an important role in the pathogenesis of systemic lupus erythematosus (SLE). In order to examine this hypothesis, we assessed the whole degree of clonal accumulation of T cells using RT-PCR and subsequent single-strand conformation polymorphism (SSCP) analysis. The analysis of the beta chain of the TCR revealed little clonotypic T cell expansion in the peripheral blood of lupus patients in remission, whereas in patients with active disease many dominant T cell clonal expansions without any distinct V beta bias were observed. The alteration in the number of T cell clones correlated well with disease activities, since these T cell expansions decreased as patients had improved. Furthermore, similar but more intense accumulations of T cell clones were found in pleural and pericardial effusions of patients with lupus serositis. Some of these identical expanded clonotypes were observed irrespective of the lesions and the times of sampling, and some of them were identical to those observed in the peripheral blood. These results suggest that the T cell clonal expansions correlate with the disease activities and that the expansion might contribute to the pathogenic lesion formation in SLE.

The distribution of TM-316-associated surface antigen on polymorphonuclear leucocytes: an immunoelectron microscopic study.

It has been established that MoAb TM-316 recognizes an epitope on leucocytes and specifically inhibits the chemotactic behavior of leucocytes. In the present paper, the distribution of this epitope on the cell surface and in intracellular organelles was studied by immunoelectron microscopy. Leucocytes separated from the blood of healthy men and from synovial fluid from patients suffering from rheumatoid arthritis were used. They were fixed with a mixture containing paraformaldehyde, glutaraldehyde and picric acid. As the second antibody, goat anti-mouse IgM conjugated to 10 nm gold colloids was employed. In normal specimens, the epitope was found to some extent on the cytoplasmic membrane of neutrophilic leucocytes, but it was only sparsely distributed on eosinophilic and basophilic leucocytes. On activated neutrophilic leucocytes, obtained from the synovial fluid of rheumatoid arthritis patients, the immunolabeling was markedly increased. The number of sites where the epitope occurs on the surface of leucocytes is thus associated with the cell type, and also with the level of activation of the leucocytes. In order to investigate the processing of the antigen, the intracellular localization of the epitope in the neutrophilic leucocytes was also studied. The epitope recognized by TM-316 was also detected in/on the characteristic granules and Golgi stacks.

Identification of commonly recognized epitopes on a rare autoantigen: poly(ADP-ribose) polymerase.

To analyze mechanisms of autoantibody production, epitope mapping of a rare autoantigen, poly(ADP-ribose) polymerase, was performed. A cDNA fragment (1873 bp long), which was already confirmed to encode the autoepitopes of this protein, was subcloned into a protein expression plasmid pEX. Several deletion mutants were produced by enzymatic treatments of this construct. PCR-amplified cDNA fragments were also individually subcloned into this vector. The recombinant proteins produced in Escherichia coli by these vectors were tested for their respective antigenicities by immunoblotting. It was found that all positive sera tested (seven cases) strongly recognized common epitopes in a restricted region of the molecule. Furthermore, three out of the seven positive sera were found to recognize other parts of the molecule. The data suggest possible mechanisms for the formation of anti-poly(ADP-ribose) polymerase autoantibodies.

Effect of the location of chewing on facial and jaw muscles activity and the mandibular movement.

This paper evaluates the influence of the location of chewing on facial and jaw muscles' activity and mandibular movements. Ten subjects with complete dentition performed following chewing sequences; 1) right side chewing (unilateral location-unspecified chewing); 2) chewing on right molars; 3) chewing on right premolars; and 4) chewing on anterior teeth. Bilateral activities of upper lip, cheek, masseter (anterior, middle and posterior parts), and anterior bellies of digastrics were recorded by surface electromyography. Mandibular movements were simultaneously recorded by electrognathography. Chewing on anterior teeth showed smaller masseteric activity in posterior part, smaller envelope of motion, and lower velocity of both opening and closing than unilateral location-unspecified chewing and chewing on right molars. Chewing on premolars showed smaller envelope of motion than chewing on molars. No significant difference was found between unilateral location-unspecified chewing and chewing on premolars or molars.

B cell epitope on the U1 snRNP-C autoantigen contains a sequence similar to that of the herpes simplex virus protein.

The mechanism of autoantibody production in autoimmune diseases is not well understood. In the present study we performed the B cell epitope mapping of the U1 small nuclear ribonucleoprotein (snRNP)-C, one of the target molecules of anti-nRNP autoantibody to investigate how B cells respond to the autoantigen. After cloning and expression of a full length complementary DNA (cDNA) encoding the U1-C protein, several truncated mutants of the cDNA were constructed and expressed in E. coli. Although a few epitopes were distributed on the whole molecule, all anti-C protein antibody-positive patients' sera tested recognized the region between amino acid residues 102 and 125 of the coding sequence. This universal epitope region contains an amino acid sequence similar to that of the herpes simplex virus type 1 ICP4 protein. The peptides representing each molecule were cross-reactive to each other. In addition this region cross-reacted to the B/B' protein. These observations suggest that molecular mimicry might be involved in the initiation of autoantibody production, followed by cross-reactive events between the autoantigens and by antigen-driven mechanisms to generate more complicated autoantibody patterns against the U1 snRNP complexes.

[A case of dermatomyositis which rapidly developed to respiratory failure in the presence of pneumomediastinum].

A 49-year-old woman was admitted in February 1987, with a six-month history of joint pain, maculopapular and erythematous rash, proximal muscle weakness and a two-month history of skin ulceration and dyspnea on exertion. Physical examination showed Gottron's papules on her fingers and a faint heliotrope rash. Biopsy of erythematous skin lesions on the shoulder and the back of the hand revealed perivascular inflammatory cell infiltration and tiny ulcerative lesions of the cutaneous tissue. Biopsy of the right quadriceps muscle showed type II fiber atrophy and slight perivascular lymphocytic infiltrate, whereas serum CPK level was within normal range. Chest X-ray film showed granular infiltrates in both lower lung fields. Based on the current findings the case was diagnosed as dermatomyositis associated with interstitial pneumonia. The administration of prednisolone, 30 mg/day resulted in the improvement of the skin lesions and muscle weakness, while the intensity of lung infiltrates was little affected. Three months after steroid therapy, the patient was readmitted because of increasing dyspnea on exertion and multiple skin ulcers. Chest X-ray revealed a small amount of gas in the mediastinum, in addition to slight deterioration of interstitial lung disease. In spite of various treatments for pneumomediastinum, including bed rest, administration of analgesics and oxygen inhalation, it developed rapidly, complicated severe subcutaneous emphysema and right-sided pneumothorax. Although high-dose prednisolone therapy and mediastinal drainage were performed, the pneumomediastinum was not resolved and she died from respiratory failure. At autopsy, predominant histological features of the lungs were acute interstitial pneumonia with hyaline membrane and edematous granulation formation in the alveoli.(ABSTRACT TRUNCATED AT 250 WORDS)

Tridimensional observation of fluorescent granular perithelial (FGP) cells in rat cerebral blood vessels.

The tridimensional appearance and distribution of FGP (fluorescent granular perithelial) cells was studied by means of light and scanning electron microscopy. In young rats they first appeared as hexagonal cells in that were closely associated; later they transformed into slender forms and were loosely arranged. Scanning electron microscope observation gave a general view of FGP cells, their globular vacuolated inclusions, and their hypertrophied protrusion into the luminal surface of blood vessels. The nodular protrusions may be related to the limitation of blood flow in small cerebral blood vessels.

An attempt to differentiate further between microglia and fluorescent granular perithelial (FGP) cells by their capacity to incorporate exogenous protein.

It seems established that under pathological conditions, microglia and blood monocytes (invading the cerebral parenchyma) behave as histiocytic cells in the central nervous system. However, it has not been clear whether or not phagocytic cells are present in normal cerebral tissue. Recently, we found a new type of cell having an uptake capacity for exogenous substance at the bifurcations of small cerebral vessels except for capillaries. According to Imamoto et al. (1982), ameboid microglia, a kind of precursor of microglia, appear at a perinatal stage and can incorporate exogenous material. In the present paper, the developmental sequences of ameboid microglia and the unique cells laden with fluorescent granules are compared at a light and electron-microscopic level. From this study, it is clear that ameboid microglia are already present in the corpus callosum at 5 days after birth and are potent in their uptake capacity for horseradish peroxidase (HRP). However, at 2 weeks, they transform into star cells and the capacity for incorporation diminishes markedly. The finding is also supported by the quantitative analysis of transformation of ameboid microglia. At 3 months, glial cells do not take the administered HRP under the present conditions. On the other hand, fluorescent granular perithelial (FGP) cells arise from a leptomeningeal tissue (pia mater) and become situated in the perivascular spaces. They are not clearly defined at 5 days, and their uptake capacity for HRP has not yet developed. At 2 weeks, the FGP cells take definite forms with several inclusion bodies, and their uptake capacity for HRP attains a certain degree. Often, they are located at bifurcations of small blood vessels. At 3 months, the FGP cells differentiate completely in appearance, and their pinocytotic capacity reaches a high level. Consequently, the FGP cells belong to a different type of cell from that of ameboid microglia in their developmental sequences and assume a principal role of scavenging waste products in normal cerebral tissue.

Distribution and number of fluorescent granular perithelial cells in coronal sections of rats cerebrum.

The fluorescent granular perithelial cells (F.G.P.) of rats aged 1 week to 2 years were observed under a light microscope to investigate intracellular granules and localization. This study showed that a marked proliferation of F.G.P. occurs within 3 weeks after birth and the total number remains constant for 2 years. The F.G.P. are mainly distributed in the gray matter, and are scarce in the white matter. The number and distribution of F.G.P. seems to reflect a difference of vascularization and function in different cerebral regions.