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1.
Chirurg ; 87(6): 469-77, 2016 Jun.
Article in German | MEDLINE | ID: mdl-27138268

ABSTRACT

BACKGROUND: A systematic approach to the etiology and possible course of acute mediastinitis is a prerequisite for adequate diagnostics and therapy. Chronic mediastinitis represents a rarity in the clinical practice. MATERIAL AND METHOD: A selective literature search was carried out. RESULTS: An acute infection of the mediastinum occurs after perforation of mediastinal structures, such as the esophagus and trachea mostly of iatrogenic origin and as descending necrotizing mediastinitis (DNM) from oropharyngeal foci. The mortality rate of esophageal injuries, irrespective of the cause is currently given as 12 %. A DNM results from an unobstructed spread along the cervicothoracic spaces and is a severe infection which manifests as a clinical picture of sepsis. The mortality rate given in the currently available literature is 14 %. Chronic mediastinitis is a very rare condition which is characterized by the proliferation of fibrous and collagenous tissue in the mediastinum. Whereas the pathogenesis remains unclear, there are indications for a Histoplasma capsulatum infection as the causal link. The prognosis is good. CONCLUSION: After perforation of the esophagus or trachea there is always the risk of an infection of the mediastinum; therefore, the diagnosis is followed by further evaluation and early therapy. The DNM can cause unspecific symptoms of sepsis without an obvious focal point. It is important to be aware of a possible correlation between an oropharyngeal center of infection and mediastinitis in order to initiate appropriate diagnostic imaging in cases with the slightest suspicion. Chronic mediastinitis is a rare condition with varying courses and can be difficult to diagnose. An histological clarification for distinction from malignant diseases appears to be a sensible approach.


Subject(s)
Mediastinitis/etiology , Mediastinitis/therapy , Acute Disease , Chronic Disease , Humans , Mediastinitis/diagnosis , Mediastinitis/mortality , Necrosis , Risk Factors , Survival Analysis , Tomography, X-Ray Computed
2.
Acta Neurol Scand ; 131(6): 381-8, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25402869

ABSTRACT

OBJECTIVES: Presence of oligoclonal bands (OCB) in cerebrospinal fluid (CSF) is a diagnostic hallmark of multiple sclerosis (MS). However, up to 10% of patients were OCB negative in routine laboratory tests. The aim of this study was to determine whether there is at least an oligoclonal restriction of intrathecal antibody synthesis against measles, rubella and/or varicella zoster virus (MRZ-specific OCB) in CSF from oligoclonal bands-negative patients with MS. METHODS: CSF and serum samples from 17 well-defined OCB-negative patients with MS were analysed for MRZ-specific OCB. We performed isoelectric focusing (IEF) combined with affinity blotting using viral antigens, detection with a highly sensitive chemiluminescence technique and recording with X-ray films. Controls included 18 OCB-positive patients with MS and 11 patients with pseudotumor cerebri (PTC). RESULTS: Exclusive or predominant MRZ-specific OCB in CSF against at least one virus species were present in 8 of 17 patients with MS (47.1%; P = 0.0422), suggesting an oligoclonal intrathecal immune response, although OCB of total IgG were absent. Only a very weak oligoclonal reaction against varicella zoster virus in CSF from one of the PTC controls was detectable. Thirteen of 18 (72.2%; P = 0.0013) OCB-positive patients with MS showed also MRZ-specific oligoclonal bands against at least 1 neurotropic virus in CSF. CONCLUSIONS: MRZ-specific OCB argue for existence of a chronic intrathecal immune reaction also in routine laboratory-OCB-negative patients with MS. This phenomenon reflects oligoclonal restriction of the humoral immunoreaction as well as polyspecific intrathecal antibody synthesis, which are both characteristics in the chronic inflammatory process of MS.


Subject(s)
Antibodies, Viral/cerebrospinal fluid , Multiple Sclerosis/immunology , Oligoclonal Bands/cerebrospinal fluid , Adult , Female , Humans , Immunoglobulin G/cerebrospinal fluid , Middle Aged , Multiple Sclerosis/cerebrospinal fluid , Rubella/immunology
3.
J Biotechnol ; 169: 51-62, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24216341

ABSTRACT

Acremonium chrysogenum is the major producer of the ß-lactam antibiotic cephalosporin C and therefore of great importance for the pharmaceutical industry. However, this filamentous fungus is known to reproduce solely by asexual means, shows only sporadic conidiospore production, and has gradual fragmentation of the vegetative mycelium into arthrospores. Due to these peculiar growth characteristics and life style, strain improvement by recombinant technologies is much more challenging than for other biotechnologically relevant fungi. Here, we describe several molecular tools for genetic engineering of A. chrysogenum, including a ΔAcku70 deletion strain for homologous recombination. No physiological or morphological changes occurred due to deletion of the ku70 gene or integration of the nat1 cassette in this recipient strain. We also used a xylose-inducible promoter from Sordaria macrospora (Smxyl) to demonstrate induction of the gfp reporter gene in A. chrysogenum. The Smxyl promoter was used for construction of a vector molecule to develop a one-step FLP/FRT recombination system in A. chrysogenum. This system was then used in the ΔAcku70 deletion strain to construct a marker-free recipient strain for targeted DNA insertion into genomic DNA. The applicability of our tools was demonstrated by construction of a marker-free transgenic strain, lacking any foreign genes.


Subject(s)
Acremonium/genetics , Anti-Bacterial Agents/metabolism , Genetic Engineering , beta-Lactams/metabolism , Acremonium/metabolism , Gene Deletion , Gene Expression Regulation , Gene Targeting , Promoter Regions, Genetic
4.
Dtsch Med Wochenschr ; 138(22): 1159-62, 2013 May.
Article in German | MEDLINE | ID: mdl-23700302

ABSTRACT

HISTORY: A 32-year-old woman was admitted to the emergency department because of acute dyspnea and syncope. A few minutes before the onset of symptoms, she had self-administered an intravenous injection of one gram of heroin combined with grinded flunitrazepam tablets. INVESTIGATIONS: Signs of acute cor pulmonale were detected on transthoracic echocardiography despite lack of pulmonary embolism in computed tomography. It was assumed that microembolisms were the cause of acute pulmonary hypertension after intravenous injection of heroin and flunitrazepam. TREATMENT AND COURSE: Because of lack of thrombus in CT scan therapeutic anticoagulation with unfractionated heparin and oxygen insufflation was initiated resulting in rapid improvement of oxygen saturation and blood pressure. On the following day pulmonary pressure in transthoracic echocardiography was already decreased significantly. Without signs of deep venous thrombosis in duplex scan and only a marginal sub segmental perfusion deficit in ventilation-perfusion-scintigraphy therapeutic anticoagulation was recommended for three months. CONCLUSION: The most likely cause of micro embolisms in this case are particles of talc, which are often used to cut heroin, or the microcrystalline cellulose used in tablets. There have been reports of tissue necrosis due to arterial embolism/vasospasm by crystalloid or oily substances (embolia cutis medicamentosa) in the extremities after intraarterial injection of grinded flunitrazepam tablets. Therefore it seems plausible that intravenous application may cause a serve but transient deficit of perfusion in pulmonary circulation.


Subject(s)
Flunitrazepam/toxicity , Heroin/poisoning , Pulmonary Heart Disease/chemically induced , Substance Abuse, Intravenous/complications , Substance Abuse, Intravenous/diagnosis , Ventricular Dysfunction, Right/chemically induced , Adult , Diagnosis, Differential , Female , Flunitrazepam/administration & dosage , Heroin/administration & dosage , Humans , Pulmonary Heart Disease/diagnosis , Pulmonary Heart Disease/therapy , Ventricular Dysfunction, Right/diagnosis , Ventricular Dysfunction, Right/therapy
5.
Ultraschall Med ; 34(5): 446-53, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23371907

ABSTRACT

PURPOSE: The conventional parameter of systolic function is global left ventricular (LV) ejection fraction (EF), but this parameter will be replaced by global strain because it seems to be more robust. However, regional strain differences can have a significant impact on global strain. Thus, the aim of the present study was to evaluate the effect of non-standardized scanning on regional strain values determined by 2D speckle tracking and tissue velocity imaging (TVI). Regional longitudinal peak systolic strain (PSS) was measured in standardized data sets of the apical 4-chamber view (ChV) and in a standardized oblique foreshortened view in patients with normal wall motion patterns. MATERIALS AND METHODS: A standardized 4ChV and a foreshortened 4ChV - defined by distinct cardiac structures - were acquired using a Vivid E9 system in 54 patients. All regional PSS values measured in monoplane 2D loops in lateral and septal regions were analyzed to detect the differences between regional strain measured in the standard and the foreshortened view. RESULTS: Significant PSS differences due to FS were detected in patients using 2D speckle tracking for the basal septal regions (p < 0.001). No significant differences due to FS were detected in patients during the analysis of TVI-based strain values (p > 0.05, paired sample T-test). CONCLUSION: To our knowledge this is the first study focusing on methodological aspects - especially standardization - using speckle tracking and TVI. Due to the lower accuracy of strain calculation based on TVI in basal regions, foreshortening has no significant impact on quantitative parameters of TVI-derived strain values in normal contracting hearts. Using speckle tracking, however, foreshortening induces significant differences of basal septal strain in normal contracting hearts. In the presence of regional wall motion defects, a lack of standardization of the views will cause inhomogeneous patterns of regional strain depending on the scan planes through the center of the infarction or its penumbra. Thus, non-standardization will have a significant impact on deformation parameters in 2D echocardiography.


Subject(s)
Echocardiography, Doppler/methods , Elasticity Imaging Techniques/methods , Image Interpretation, Computer-Assisted/methods , Systole/physiology , Ventricular Dysfunction, Left/diagnostic imaging , Ventricular Dysfunction, Left/physiopathology , Adult , Aged , Aged, 80 and over , Cardiac Volume/physiology , Diastole/physiology , Echocardiography, Doppler, Color/methods , Female , Humans , Hypertension/complications , Hypertension/diagnostic imaging , Male , Middle Aged , Reference Values
6.
Acta Biomater ; 8(4): 1530-42, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22202909

ABSTRACT

An important challenge in the biomaterials field is to mimic the structure of functional tissues via cell and extracellular matrix (ECM) alignment and anisotropy. Toward this goal, silk-based scaffolds resembling bone lamellar structure were developed using a freeze-drying technique. The structure could be controlled directly by solute concentration and freezing parameters, resulting in lamellar scaffolds with regular morphology. Different post-treatments, such as methanol, water annealing and steam sterilization, were investigated to induce water stability. The resulting structures exhibited significant differences in terms of morphological integrity, structure and mechanical properties. The lamellar thicknesses were ∼2.6 µm for the methanol-treated scaffolds and ∼5.8 µm for water-annealed. These values are in the range of those reported for human lamellar bone. Human bone marrow-derived mesenchymal stem cells (hMSC) were seeded on these silk fibroin lamellar scaffolds and grown under osteogenic conditions to assess the effect of the microstructure on cell behavior. Collagen in the newly deposited ECM was found aligned along the lamellar architectures. In the case of methanol-treated lamellar structures, the hMSC were able to migrate into the interior of the scaffolds, producing a multilamellar hybrid construct. The present morphology constitutes a useful pattern onto which hMSC cells attach and proliferate for guided formation of a highly oriented extracellular matrix.


Subject(s)
Guided Tissue Regeneration/methods , Silk/chemistry , Tissue Engineering/methods , Absorption , Alkaline Phosphatase/metabolism , Amides/chemistry , Animals , Cell Shape , Cells, Cultured , Crystallization , DNA/metabolism , Fibroins/chemistry , Fibroins/ultrastructure , Freeze Drying , Humans , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/enzymology , Mesenchymal Stem Cells/ultrastructure , Microscopy, Confocal , Microscopy, Electron, Scanning , Silk/ultrastructure , Spectroscopy, Fourier Transform Infrared , Tissue Scaffolds , Vibration , Water/chemistry
7.
J Cardiovasc Dis Res ; 2(1): 68-70, 2011 Jan.
Article in English | MEDLINE | ID: mdl-21716756

ABSTRACT

The sticky platelet syndrome (SPS) is a congenital disorder characterized by platelet hyperaggregability to epinephrine and/or adenosine diphosphate; this predisposes affected individuals to acute myocardial infarction, ischemic optic neuropathy, recurrent venous thromboembolism, and transient ischemic cerebral attacks and strokes. Here, we describe an unusual case with recurrent cerebrovascular accidents due to SPS, in the presence of a patent foramen ovale (PFO). We report an unusual case of a 56-year-old female patient with a PFO, who suffered from recurrent strokes despite long-term medication with clopidogrel for SPS. The patient underwent successful transcatheter closure of the PFO, and, in addition, she has been placed on low-dose acetylsalicylic acid. After 18-month follow-up, she demonstrated an intact atrial septum without any vegetations on the percutaneous device until today. She has had no further thromboembolic events.

8.
Internist (Berl) ; 51(12): 1567-70, 2010 Dec.
Article in German | MEDLINE | ID: mdl-21069275

ABSTRACT

We report a case of a 35 year old male with severe deep vein thrombosis of the lower limb on both sides and pulmonary embolism. A Klinefelter's mosaic (47,XXY [81%]/48,XXXY [19%]) was diagnosed. Because no other cause for this thromboembolism was found, we assume that in part, it was caused by the Klinefelter's mosaic. In all male patients presenting with thromboembolism, especially those with an unusual habitus, a Klinefelter's syndrome should be considered as differential diagnosis. Testosterone substitution therapy should be started in all patients with Klinefelter's syndrome to prevent further disease.


Subject(s)
Klinefelter Syndrome/diagnosis , Pulmonary Embolism/etiology , Venous Thrombosis/etiology , Adult , Anticoagulants/therapeutic use , Blood Coagulation Tests , Combined Modality Therapy , Diagnosis, Differential , Humans , In Situ Hybridization, Fluorescence , Karyotyping , Klinefelter Syndrome/genetics , Male , Mosaicism , Phenprocoumon/therapeutic use , Pulmonary Embolism/drug therapy , Pulmonary Embolism/genetics , Stockings, Compression , Tomography, X-Ray Computed , Ultrasonography, Doppler, Duplex , Venous Thrombosis/drug therapy , Venous Thrombosis/genetics
9.
Plant Biol (Stuttg) ; 12(5): 788-93, 2010 Sep 01.
Article in English | MEDLINE | ID: mdl-20701702

ABSTRACT

The colonisation of land by plants may not have been possible without mycorrhizae, which supply the majority of land plants with nutrients, water and other benefits. In this sense, the mycorrhization of basal groups of land plants such as ferns and lycophytes is of particular interest, yet only about 9% of fern and lycophyte species have been sampled for their mycorrhization status, and no community-level analyses exist for tropical fern communities. In the present study, we screened 170 specimens of ferns and lycophytes from Malaysia and Sulawesi (Indonesia), representing 126 species, and report the mycorrhization status for 109 species and 19 genera for the first time. Mycorrhizal colonisations were detected in 96 (56.5%) of the specimens, 85 of which corresponded to arbuscular mycorrhizae (AMF), three to dark-septate endophytes (DSE) and four to mixed colonisations (AMF + DSE). DSE colonisations were lower than in comparable samples of ferns from the Andes, suggesting a geographical or taxonomic pattern in this type of colonisation. Epiphytes had significantly lower levels of colonisation (26.1%) than terrestrial plants (70.7%), probably due to the difficulty of establishment of mycorrhizal fungi in the canopy habitat.


Subject(s)
Ecosystem , Ferns/microbiology , Mycorrhizae/classification , Indonesia , Malaysia , Plant Roots/microbiology
11.
Zentralbl Chir ; 127(6): 533-7, 2002 Jun.
Article in German | MEDLINE | ID: mdl-12094281

ABSTRACT

The importance of laparoscopy in the management of blunt abdominal trauma should be evaluated. Therefore we retrospectively analysed all patients with blunt abdominal trauma treated in the Department of Surgery at the Carl-Thiem-Hospital Cottbus between 1998 and 2000. Within this period a total number of 53 patients with blunt abdominal trauma underwent operative treatment, 20 (37.7 %) of them had primary laparoscopy. Of the 11 cases where laparoscopic operation could be completed without conversion to exploratory laparotomy, 8 patients had intra-abdominal injuries and underwent sufficient laparoscopic treatment. The percentage of so called "negative" exploratory laparotomies within this study was 13.2 %. Our analysis suggests that laparoscopy should become firmly established in the diagnostic management and, if indicated, in the treatment of blunt abdominal trauma as well.


Subject(s)
Abdominal Injuries/surgery , Laparoscopy , Wounds, Nonpenetrating/surgery , Abdominal Injuries/diagnosis , Abdominal Injuries/mortality , Adolescent , Adult , Child , Female , Hemoperitoneum/etiology , Hemoperitoneum/mortality , Hemoperitoneum/surgery , Humans , Liver/injuries , Liver/surgery , Male , Mesentery/injuries , Mesentery/surgery , Middle Aged , Multiple Trauma/mortality , Multiple Trauma/surgery , Retrospective Studies , Splenic Rupture/diagnosis , Splenic Rupture/mortality , Splenic Rupture/surgery , Survival Rate , Wounds, Nonpenetrating/diagnosis , Wounds, Nonpenetrating/mortality
12.
J Am Vet Med Assoc ; 215(4): 511-4, 483, 1999 Aug 15.
Article in English | MEDLINE | ID: mdl-10461636

ABSTRACT

A 7-month-old foal was admitted to the hospital with a history of lethargy, weight loss, mild diarrhea, and anorexia. A diagnosis of proliferative enteritis caused by Lawsonia intracellularis-like organisms was made after necropsy and histologic examination of the small intestine. Although infection with L intracellularis-like organisms is a rare cause of enteritis in foals, it should be considered in the differential diagnosis, especially if the foal was housed in the proximity of pigs or pig feces. Antemortem diagnosis remains challenging because isolation of the organism in fecal material requires cell culture, and histologic evaluation of intestinal biopsy specimens may be unrewarding because of the lack of information regarding the frequency and distribution of lesions in horses. Alternatively, use of immunochemical stain, dot-blot technique, and polymerase chain reaction provide specific diagnostic tests that can be performed on fecal material. Postmortem diagnosis relies on histologic examination of infected tissues and use of immunofluorescence and polymerase chain reaction.


Subject(s)
Gastroenteritis/veterinary , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacterial Infections/veterinary , Horse Diseases/physiopathology , Animals , Antibodies, Monoclonal , Diagnosis, Differential , Fatal Outcome , Female , Gastroenteritis/diagnosis , Gastroenteritis/microbiology , Gastroenteritis/physiopathology , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/physiopathology , Horse Diseases/diagnosis , Horse Diseases/microbiology , Horses , Immunohistochemistry , Intestine, Small/pathology , Lung/pathology , Microscopy, Electron/veterinary , Microscopy, Fluorescence/veterinary
13.
Zentralbl Chir ; 124(1): 63-7, 1999.
Article in German | MEDLINE | ID: mdl-10091302

ABSTRACT

Based on animal trials the presented study describes two versions of thoracoscopic oesophageal anastomosis within the scope of abdomino-thoracoscopic oesophagectomy. This experimental approach is considered to provide a solution for the oncological problem to salvage the tumor bearing oesophagus. We describe the procedure of intrathoracic stapler anastomosis under thoracoscopic vision. By using a laparoscopic purse string suture clamp we were enabled to prepare the proximal oesophageal stump for anastomosis. A circular stapling instrument turned out to be very suitable for the thoracoscopic use. The final evaluation of the importance of minimally invasive surgery for the resection of oesophageal carcinomas should not be given until multicenter studies are performed.


Subject(s)
Anastomosis, Surgical/instrumentation , Endoscopes , Esophagectomy/instrumentation , Thoracoscopes , Animals , Male , Minimally Invasive Surgical Procedures/instrumentation , Surgical Instruments , Surgical Staplers , Suture Techniques/instrumentation , Swine
14.
Zentralbl Chir ; 123(8): 944-9, 1998.
Article in German | MEDLINE | ID: mdl-9757540

ABSTRACT

Based on animal trials the presented study results describe a laparoscopic assisted removal of an extra-long free jejunal graft to reconstruct voice and swallowing after a total laryngo-pharyngectomy in cases with advanced malignancies of the larynx including invasion in the upper oesophagus. With a microvascular anastomosed small bowel segment, we have been able to reconstruct parts of the pharynx and to create a speaking syphon as devised by Ehrenberger in an one-stage procedure. The concept of avoidance of an extensive laparotomy may result in an decreased morbidity of this surgical procedure. Therefore it represents a challenging single indication of minimally invasive surgery of the oesophagus. A well established interdisciplinary teamwork is the key for success in this extensive procedure.


Subject(s)
Jejunum/transplantation , Laparoscopes , Laryngectomy/instrumentation , Pharyngectomy/instrumentation , Speech, Alaryngeal , Surgical Flaps , Animals , Male , Swine
15.
J Head Trauma Rehabil ; 13(2): 27-35, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9575254

ABSTRACT

OBJECTIVE: This study investigated the clinical utility of the Standardized Assessment of Concussion (SAC) in detecting concussion in athletes. METHOD: Athletic trainers administered the SAC to 568 nonconcussed high school and college football players prior to the 1995 and 1996 football seasons. Thirty-three of these players experienced concussion and were tested immediately following injury; 28 of the 33 underwent additional follow-up testing 48 hours after the injury. RESULTS: Concussed players scored significantly below nonconcussed controls on all SAC measures and significantly below their own pre-injury baseline performance. Follow-up testing documented return to preinjury baseline. CONCLUSION: These findings support the SAC's effectiveness in detecting concussion and tracking recovery in order to determine a player's fitness to return to play.


Subject(s)
Athletic Injuries/diagnosis , Brain Concussion/diagnosis , Neuropsychological Tests , Adolescent , Adult , Humans , Male , Reproducibility of Results
16.
Zentralbl Chir ; 123(2): 188-92, 1998.
Article in German | MEDLINE | ID: mdl-9556894

ABSTRACT

Based on animal trials the presented study results describe a method of abdomino-thoracoscopic oesophagectomy for the treatment of oesophageal carcinoma of the middle and lower third. Technical details and the advantages of the surgical procedure, including the avoidance of thoracotomy and the practicability of the thoracoscopic dissection of the oesophagus are shown. Problems exist in the salvage of the specimen, especially in the transcervical passage of the tumorbearing oesophageal part. A final evaluation of the importance of the minimally invasive techniques in oesophageal surgery should not be given until the results of clinical studies are available.


Subject(s)
Endoscopes , Esophagectomy/instrumentation , Laparoscopes , Thoracoscopes , Anastomosis, Surgical/instrumentation , Animals , Male , Surgical Instruments , Suture Techniques/instrumentation , Swine
17.
Arch Virol ; 143(12): 2471-85, 1998.
Article in English | MEDLINE | ID: mdl-9930203

ABSTRACT

Transmissible gastroenteritis virus (TGEV) is a coronavirus which causes severe gastroenteritis and atrophy of intestinal villous epithelial cells in piglets. However, the mechanism of cell death caused by TGEV is not known. In this study, we report that TGEV induces cell death by apoptosis. TGEV-induced apoptosis was demonstrated by agarose gel electrophoresis, electron microscopy, and terminal deoxytransferase digoxigenin-dUTP nick end labeling (TUNEL). Double labeling experiment confirmed the result from electron microscopy and showed that most of the apoptotic cells were bystander cells as they were negative for TGEV nucleic acids. Results of this study indicate that TGEV induces apoptosis in vitro and that most of the cells undergoing apoptosis are bystander cells, thus amplifying the cytopathic effect of TGEV.


Subject(s)
Apoptosis , Transmissible gastroenteritis virus/pathogenicity , Animals , Cells, Cultured , Cytopathogenic Effect, Viral , DNA Fragmentation , In Situ Hybridization , In Situ Nick-End Labeling , Male , Microscopy, Electron , Swine , Testis/cytology , Transmissible gastroenteritis virus/genetics , Transmissible gastroenteritis virus/ultrastructure
18.
J Gen Virol ; 79 ( Pt 12): 2989-95, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9880013

ABSTRACT

Evidence of apoptosis was detected for the United States porcine reproductive and respiratory syndrome virus (PRRSV) in ATCC CRL11171 cells inoculated with strain ATCC VR2385 and in the tissues of pigs infected with the same strain. Apoptosis was detected by agarose gel electrophoresis, transmission electron microscopy and terminal deoxytransferase dUTP nick end labelling (TUNEL) techniques. By electron microscopy and double-labelling techniques, apoptosis was detected primarily in uninfected bystander cells in the continuous cell line rather than the PRRSV-infected cells. In the lungs, the apoptotic cells were predominantly alveolar and pulmonary intravascular macrophages, and mononuclear cells in the alveolar septa. In the lymph nodes, the apoptotic cells were predominantly tingible body macrophages and mononuclear cells. The induction of apoptosis in a large number of mononuclear cells in the lungs and lymph nodes appears to be a mechanism of PRRSV pathogenesis and might be an explanation for a dramatic reduction in the number of alveolar macrophages and circulating lymphocytes and monocytes in PRRSV-infected pigs.


Subject(s)
Apoptosis , Porcine respiratory and reproductive syndrome virus/physiology , Animals , Porcine Reproductive and Respiratory Syndrome/pathology , Porcine Reproductive and Respiratory Syndrome/virology , Swine , United States , Virulence
19.
Vet Microbiol ; 57(2-3): 283-9, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9355262

ABSTRACT

The pathology of bovine herpesvirus type 4 (BHV-4) infection was studied in cats, rabbits and guinea pigs. Twenty kittens, twenty-two rabbits and ten guinea pigs, some treated with glucocorticoid-were inoculated with a BHV-4 strain of feline origin, via various routes of inoculation (conjunctival, intranasal, peritoneal). Clinical signs were recorded. After euthanizing at different post inoculation days macro- and microscopic changes were observed by necropsy and in hematoxylin-eosin stained histological sections. The presence of the virus in organs was detected by immunohistochemistry and a nested PCR assay. Inclusion bodies and monoclonal antibody-stained cells were found in the conjunctiva, trachea, lungs, spleen and lymph nodes. Most of the lesions were localized to the respiratory and the immune system. The macro- and microscopic lesions and clinical signs were more severe in kittens and guinea pigs. The histological data indicated that cats, especially kittens, were susceptible for BHV-4 and the infection was not confined to the urinary bladder.


Subject(s)
Gammaherpesvirinae , Herpesviridae Infections/veterinary , Animals , Cat Diseases , Cats , Cattle , Glucocorticoids/pharmacology , Guinea Pigs , Herpesviridae Infections/mortality , Herpesviridae Infections/pathology , Polymerase Chain Reaction , Prednisolone/pharmacology , Rabbits , Species Specificity
20.
Mol Cell Probes ; 11(4): 273-80, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9281413

ABSTRACT

In situ hybridization (ISH) is a useful diagnostic and research tool, but is also time consuming. This study was conducted to determine if a rate enhancement hybridization (REH) buffer, developed for membrane hybridization, could be used to decrease hybridization time for ISH. Tissue from swine with an enteric disease produced by a swine coronavirus, transmissible gastroenteritis virus (TGEV), was used as a model to standardize hybridization conditions for a rapid ISH technique. Small intestinal sections from pigs experimentally and naturally infected with TGEV were hybridized for various times at 52 degrees C and 70 degrees C with a radiolabelled or a fluorescein-labelled RNA probe in a standard hybridization or a REH buffer. Viral RNA was detected in intestines from as early as 30 min of hybridization by using both buffers with the radiolabelled probe; however, the signal was stronger with the REH buffer. With the fluorescein-labelled probe, viral RNA was detected in virus-infected cells of the intestines after 30 min of hybridization by using the REH buffer. Signal intensity was greater with the REH buffer than with the standard hybridization buffer when compared at each hybridization time and hybridization temperature using both radiolabelled and fluorescein-labelled probes. With the REH buffer, hybridization signal intensity was greater at 70 degrees C than at 52 degrees C for both probes. The best results were obtained when small intestinal sections were hybridized at 70 degrees C for 2 h using a radiolabelled or a fluorescein-labelled probe diluted in the REH buffer. The fluorescein-labelled RNA probe with REH buffer resulted in a minimal non-specific signal when compared with the radiolabelled probe. These studies demonstrated that the REH buffer can be used to decrease the time of ISH for the detection of viral RNA. This rapid ISH technique should have broad applications in the utilization of probe technology in diagnostics and research for the detection of target ribonucleic acids in situ


Subject(s)
Fluorescein/chemistry , In Situ Hybridization/methods , Ribonucleotides/analysis , Animals , Buffers , Gastroenteritis/virology , Intestines/pathology , Intestines/virology , RNA/chemistry , Swine , Temperature , Transmissible gastroenteritis virus/genetics , Transmissible gastroenteritis virus/isolation & purification
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