Relationship between chicken proventricular necrosis virus prevalence and transmissible viral proventriculitis in broiler chickens in Poland.

Transmissible Viral Proventriculitis (TVP) is a disease of chickens which contributes to significant production losses. Recent reports indicate the role of chicken proventricular necrosis virus (CPNV) in the development of TVP. However, the relationship between CPNV and TVP is inconclusive and it has been addressed in just a few reports. Given the above, a study was conducted to identify the relationship between TVP and CPNV prevalence in broiler chickens in Poland. The study was carried out on 35 proventriculi samples sent for histopathological (HP) examination to the Faculty of Veterinary Medicine in Olsztyn between 2017 and 2019. After HP examination, TVP positive samples were processed for CPNV identification by RT-PCR. TVP was the most common pathological condition of proventriculi (23 cases). CPNV was identified in 10 out of those 23 cases. The average HP score, and the average necrosis and infiltration score for CPNV-positive samples was significantly higher than in CPNV-negative ones. The average age of the CPNV-positive chickens was significantly lower than in CPNV-negative birds. Our study confirms the role of CPNV in TVP pathogenesis and it seems that preservation of the proventriculi in the early stages of the disease, when the lesions are more pronounced, should result in a greater probability of CPNV detection.

An evaluation of the impact of aloe vera and licorice extracts on the course of experimental pigeon paramyxovirus type 1 infection in pigeons.

The progressive decrease in the efficiency of synthetic drugs has prompted research into phytogenic feed additives with potentially immunomodulatory and anti-infective properties. Complex diseases with a mixed etiology, including viral, pose a growing problem in domestic pigeons. The aim of this study was to determine the effectiveness of various doses of aloe vera and licorice extracts on the course of experimental PPMV-1 infection in pigeons. The experiment was performed on pigeons divided into 5 groups, including one control group and 4 experimental groups, which were orally administered aloe vera or licorice extracts at 300 or 500 mg/kg BW for 7 d after experimental inoculation with PPMV-1. On d 4, 7, and 14 after inoculation, cloacal swabs and samples of organs were collected from 4 birds in each group. The samples were analyzed to determine the copy number of PPMV-1 RNA by TaqMan qPCR. The results indicate that licorice and aloe vera extracts inhibited PPMV-1 replication by decreasing viral RNA copy numbers in the examined organs. The most inhibitory effect was observed in pigeons receiving aloe vera extract at 300 mg/kg BW, for which PPMV-1 RNA copy numbers were approximately 7-fold lower (brain), 9-fold lower (kidneys), and 14-fold lower (liver) than in the control group. The results of this study point to the potentially antiviral effects of aloe vera and licorice extracts in pigeons infected with PPMV-1. To the best of our knowledge, this is the first study to investigate the antiviral properties of aloe vera and licorice extracts in domestic pigeons.

Differentiation of infectious bronchitis virus vaccine strains Ma5 and 4/91 by TaqMan real-time PCR.

The aim of this study was to develop rapid molecular assays for differentiating vaccine strains Ma5 and 4/91 of the infectious bronchitis virus (IBV). Specific primers and probes for S1 and N genes were designed based on the nucleotide sequences of both vaccine strains. Cross-reactivity was not observed. Assay sensitivity was 2.373 × 103 copies of the Ma5 strain, and 3.852 x 103 copies of the 4/91 strain. Samples belonging to a known genotype demonstrated that the designed assays supported rapid and sensitive detection of Ma5 and 4/91 vaccine strains of IBV.

Is the increased expression of ubiquitin in CADASIL syndrome a manifestation of aberrant endocytosis in the vascular smooth muscle cells?

Ubiquitin is a highly conserved protein involved in many important cellular processes, such as cell surface receptor signaling, endocytosis and protein degradation. Since ubiquitin plays a key role in the pathomechanisms of many neurodegenerative diseases, we immunohistochemically analyzed its expression in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). CADASIL is a heritable vascular dementia characterized by degeneration of the vascular smooth muscle cells (VSMC) caused by mutations in the notch 3 gene. In CADASIL, there is abnormal accumulation of the Notch 3 extracellular domain on blood vessels, but the molecular pathways linking notch 3 mutations to degeneration of the VSMC are, as yet, poorly understood. We studied human brain and skin biopsy specimens, and observed increased ubiquitin expression on structures primarily affected by the pathological process in CADASIL: the VSMC and vascular lamina media, and also large ballooned macrophages. Ultrastructurally, we noted that pathognomonic CADASIL deposits of granular osmiophilic material were often located inside indentations in the VSMC membrane that resembled endocytic vesicles. We suggest that in CADASIL, damage to the VSMC may be associated with aberrant ubiquitin-dependent endocytosis of the Notch 3 ligand, and increased accumulation of ubiquitin on the vessel wall may be a manifestation of this aberration.

Extensive mixed vascular malformation clinically imitating multiple sclerosis--case report.

Vascular malformations usually develop as a result of influence of teratogenic factor(s) acting in the defined embryonic/fetal period. However, in the case examined by us, various types of vascular malformations formed in different periods of the ontogenic development were found. They were seen in all parts of the central nervous system and clinically mimicked multiple sclerosis. On the background of generalized ischemic lesions of the CNS, certain kinds of vascular malformations were seen: cavernous or fetallike vessels within meninges, superficially located capillary angioma penetrating into the brain and spinal cord white matter, and arterio-venous pathological conglomerates forming meningeal angiomatosis. In pathological vessels, immunocytochemical assessment of vascular endothelium with antibodies against antigens CD31, CD34, von Willebrand factor and lectin Ulex europaeus was normal but examination of the vascular basal membrane compounds revealed poor immunoreactivity to laminin and fibronectin. There were no disturbances in expression of angiopoietin, platelet-derived growth factor, transforming growth factor beta and vascular endothelial growth factor receptors Tie-1/2, PDGFR-alpha/beta, endoglin and Flk-1, respectively. The presence of various types of pathological vessels originating from different ontogenic periods indicates remittent or prolonged influence of teratogenic factor(s) in all periods of fetal vessel development.

CADASIL: new cases and new questions.

We described the first two unrelated Polish families with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). In the morphological examination with light microscopy, two kinds of changes were observed: (1). panarteritis nodosa-like changes with eosinophilic fibrinoid necrosis of the vessel wall and perivascular inflammatory infiltrates and (2). basophilic granular material in the tunica media characteristic of CADASIL. At electron microscopy, we found deposits of granular osmophilic material (GOM) within the wall of arteries, veins and capillary vessels. Our findings imply two questions requiring further investigation: Why in the genetically determined vascular disorder are the features of systemic inflammatory vascular disease present? Why in capillary walls deprived of smooth muscle cells are deposits of GOM present?

Cellular expression of tumor necrosis factor a and its receptors in human ischemic stroke.

We evaluated by immunocytochemistry cellular localization and time-dependent expression of tumor necrosis factor a (TNF-alpha) and its receptors p55 (TNF-RI) and p75 (TNF-R2) in human ischemic brains. We observed them in microglia, neurons, astrocytes, macrophages and blood vessels. Since TNF-alpha expression was very intense and prolonged in microglia, it probably constitutes the main cellular source of the cytokine following cerebral ischemia in humans. Constitutive expression of TNF-alpha receptors was observed in neurons and blood vessels while in other cells it was induced by ischemia. In macrophages, dominant immunolabeling for TNF-R2 was seen. In other cells, immunoreactions for both types of TNF-alpha receptors were similar but the pattern of immunostaining was different: homogenous for TNF-R1 and granular for TNF-R2. Beneficial and detrimental role of TNF-alpha in cerebral ischemia and supposed mechanisms of action are discussed.

Early ontogenic disturbances in cell migration in mentally disabled adult.

Disturbances in cell migration are heterogenic disorders of brain development commonly associated with epilepsy and mental retardation. We report a 45-year-old oligophrenic man with defect of lower limbs and family history of mental retardation who died because of brainstem hemorrhage. At post-mortem and histopathological examination, complex brain malformation characterized by bilateral periventricular heterotopia, cortical dysgenesia, partial agenesia of corpus callosum and thin-walled blood vessels was found. Immunohistochemical examination revealed the presence of fibronectin, collagen IV and laminin in wall of pathological vessels. Cyclooxygenase-2 (COX-2) expression in neurons within heterotopias and dysgenic cortex was negative. It may indicate their maturity and indirectly, normal activity of postsynaptic NMDA receptors that explains absence of epileptic attacks in our patient. The presence of COX-2-negative neurons and compounds of basal lamina in fetal-like vessels within heterotopias and dysgenic cortex, suggests that these probably initially immature structures became mature during 45 years of patient's life.

Paraneoplastic syndrome in the course of lung adenocarcinoma: morphological picture and immunohistochemical analysis of the inflammatory infiltrates and PECAM-1 expression.

We examined sections of brain, spinal cord, spinal roots, and peripheral nerves from a patient with paraneoplastic syndrome in the course of lung adenocarcinoma. Morphological examination showed marked loss of myelin fibers in peripheral nerves, severe brain tissue edema, and paraneoplastic degeneration involving cerebrum and cerebellum with inflammatory components. Inflammatory infiltrates examined immunohistochemically using antibodies against antigens CD 3, CD 4, CD 8, and CD 20 turned out to be composed of cytotoxic T lymphocytes. The expression of platelet-endothelial cell adhesion molecule-1 (PECAM-1) in blood vessels was increased in comparison with control material, which may facilitate transendothelial lymphocyte migration triggering a cascade of biochemical and morphological reactions observed in paraneoplastic syndrome.

Astrogliosis and blood vessel development during human spinal cord myelination.

The aim of our study was to investigate a relationship between vessel development, astrogliosis and myelination in humans. We examined spinal cords of human fetuses and infants using immunohistochemical methods with antibodies against Myelin Basic Protein (MBP), Glial Fibrillary Acidic Protein (GFAP) and lectin Ulex Europaeus Type 1 (UEA-1). Our investigation showed that in parallel to the increase of MBP reactivity in the spinal cord white matter more GFAP-immunoreactive astrocytes appeared. Reaction of vessel endothelium with lectin UEA-1 revealed the temporal increase of vascularization in the course of the spinal cord tracts myelination in fetuses. In the postnatal period when the myelination is nearly morphologically completed, the spinal cord white matter was poorly vascularized although GFAP-immunoreactive cells were still relatively numerous. We suggested that the increase of vascularization and astrogliosis observed during spinal cord tracts myelination may be connected with their participation in process of myelin sheath formation via, among others, local secretion of growth factors necessary for normal myelin development.

Correlative ultrastructural and immunohistochemical study of developing vascular basement membrane in postnatal rat spinal cord.

We investigated the development of vascular basement membrane in immature spinal cord vessels during rat spinal cord myelination. Correlative ultrastructural and immunohistochemical study indicated that fibronectin was the first component of extracellular matrix. Then, on the 9th postnatal day, laminin appeared. At that time, lamina lucida of vascular basement membrane was not detectable. On the 15th day, when collagen IV was visible, lamina densa and lamina lucida were occasionally observed. All components of basement membrane--fibronectin, laminin, collagen IV, alpha-2 laminin (merosin)--and ultrastructural division into two layers were detected on the 25th postnatal day. The results of this study indicates that a gradual development of endothelium in immature rat spinal cord blood vessels leads to a gradual increase of synthesis of extracellular matrix components.

Do astrocytes participate in rat spinal cord myelination?

Astrocytes play an important role in CNS development phenomena, such as neuron migration and blood-brain barrier formation, but only a little is known of their role in the process of myelination. The aim of our investigation was to examine the relationship between astrocytes and myelin formation. We evaluated rat spinal cords using hematoxylin-eosin and Klüver-Barrera staining methods as well as immunohistochemical methods with antibodies against myelin basic protein (MBP), glial fibrillary acidic protein (GFAP) and lamins A/C and B2. Our investigation revealed that myelination in the rat spinal cord tracts began between the 6th and 9th postnatal day involving the anterior funiculi, then the lateral funiculi and later the posterior ones. The process of myelination finished about the 25th postnatal day. More GFAP immunoreactive astrocytes were detected in parallel to the increase of MBP reactivity. We suggest that the temporary increase of GFAP positive cells accompanying the process of myelination is necessary for normal myelin development and may be connected with local secretion of growth factors by astrocytes.

Vascular changes in tuberculous meningoencephalitis.

Our report refers two cases of tuberculous encephalomeningitis which differ in the course and pathological changes. In case 1 blood vessels showed features of peri, endo-, or panvasculites. In some vessels endothelium proliferation leading to the stenosis or obliteration of the vascular lumen was observed. Necrosis was an effect of vessels occlusion. In case 2 many fewer vessel were involved in onflammation process. Vascular changes were also less extensive and were observed more rarely. Tuberculous infection often caused less tissue lesions than vascular changes. Different pathological changes probably depend on the type and virulence of Myobacterium tuberculosis and on the host immune response to the infection.

Age-dependent changes in astroglial reactivity in human ischemic stroke. Immunohistochemical study.

Clinical investigations revealed that a course of human ischemic stroke may be different in various patient's age. For the purpose of finding a reflection of this phenomenon in morphological picture we examined astroglial reactions in human brains with ischemic necrosis. Material was divided into two groups: senile (SG) (80-101 years, n = 34) and the middle age (MG) (40-65 years, n = 33). Astrocyte reactivity was evaluated using immunohistochemical methods with antibodies against: protein S-100, GFAP, Vimentin and recombinant parts of the Ki-67 molecule (MIB-1 antibody) during 32 days after the disease onset. Investigations revealed that astroglial reactivity appeared later (5th day) in SG then in MD (3rd day), was less pronounced-especially in the third (p < 0.05) and fourth week (p < 0.01), was more diffused and of shorter duration in SG than in MG. Astrocyte proliferation turned out surprisingly minimal in both groups and was observed more often in MG then in SG. We suggest that astroglial reactivity depends on the patient's age. Difference in the astrocyte reactions may be caused by a decrease of mitogenic and transforming factors activity, a decrease of astrocyte susceptibility for stimulating factors and/or ischemic tolerance phenomenon.

[Nocturnal paroxysmal hemoglobinuria--case report]. / Nocna napadowa hemoglobinuria--opis dwóch przypadków.

Paroxysmal nocturnal hemoglobinuria (PNH)-a case report. We present clinical and postmortem findings in patient with PNH who developed cerebral thrombosis. The pathogenesis of the PNH is discussed.

White matter injury in amyotrophic lateral sclerosis (ALS).

Microscopic examination of 6 brains and spinal cords of patients deceased of amyotrophic lateral sclerosis reveals as follows: 1) White matter changes considerably exceeding those found in motor neurons structures. 2) They involve white matter of frontal, temporal and parietal lobes including long association fibers. 3) Pyramidal tracts are extremely damaged within spinal cord and medulla. 4) Within spinal cord besides pyramidal tracts majority of ascending and descending pathways are damaged. 5) Pathomechanism of white matter changes seems to be complex. Within brain hemispheres predominate nonspecific changes, 6) Imaging methods in ALS bring a diagnostic progress but at present the conclusions on specificity of changes encountered in various imaging methods in different diseases seem to be precautions.

[Cerebral thrombosis in paroxysmal nocturnal hemoglobinuria: case report]. / Przypadek zakrzepicy naczyn mózgowych w przebiegu nocnej napadowej hemoglobinurii.

Clinical, radiological and postmortem findings in a 25-year old woman with paroxysmal nocturnal hemoglobinuria (PNH) who developed cerebral thrombosis are presented. The pathogenesis of the PNH is discussed.

Coexistence of various vascular malformations within the brain.

Authors present two cases of basilar artery aneurysm accompanied by different development failures of blood vessels. In both cases anomaly in formation of brain base vessels, angioma consisted of different size thin-walled vessels and arterio-venous angioma within brain stem were stated. Besides, conglomerates of abnormal vessels, angiosis within pia matter, diffused lacunar and fetal as well as thin-walled venous vessels were found. Pathological vessels, their conglomerates were present in brain stem, cerebellum and cerebral hemispheres. The variability of vascular malformations seems to point at long-lasting action pathogenic factor during ontogenesis. Authors try to refer particular developmental anomalies to proper stage of ontogenesis.

Morphological picture in paroxysmal nocturnal hemoglobinuria. Case report.

25-year-old woman with paroxysmal nocturnal hemoglobinuria was admitted to the hospital because of headache, progressing right hemiparesis and speech disorders. Several days later patient lost consciousness. Cerebrospinal fluid was xanthochromic with increased pleocytosis and protein level. CT-scan revealed ischemic area with hemorrhagic focus within left cerebral hemisphere. Patient died 3 weeks after the admission. Brain section revealed hemorrhagic infarct in the cortex of the left parietal lobe, thrombosis of the superior sagittal sinus and "respiratory brain" changes. Microscopic examination revealed meningeal venous thrombosis, hemorrhagic infarct, vasculitis, abundant accumulation of bacteria within blood vessels, and other pathological changes such as petechiae, perivascular exudates and small, round areas composed of acellular fibrillary network. There were no macrophages and GFAP-positive astrocytes in any of these areas. Authors suggest that weak cell reactivity may be connected with alterations in cell membranes, mainly low phosphatidylinositol (GPI) content.

Ammon's horn changes in focal brain ischemia in humans.

Neuronal changes in Ammon's horn were examined immunocytochemically in 20 patients aged from 51 to 101 years, deceased in the course of ischemic lesions localized within the area supplied by vessels derived from other then Ammon's horn vascularization (middle cerebral artery). Numerous neurons within various sector of the pyramidal layer, in the dentate gyrus, subiculum and entorhinal cortex were immunopositive in reaction with antibodies to serum proteins (albumin, IgG, alpha-1-antitrypsin), indicating their damage. The distribution of damaged Ammon's horn pyramidal cells differed from the location of injured Ammon's horn neurons in experimental investigations of brain ischemia and did not indicate a selective vulnerability of pyramidal cells in the human h1 area, corresponding to the CA1 sector in animals. Contrary to experimental material, changes in the human Ammon's horn are caused by numerous overlapping factors.