Biomedical subjects
H Lutz
Publications and source records attributed to H Lutz.
Tumor necrosis factor alpha levels in cats experimentally infected with feline immunodeficiency virus: effects of immunization and feline leukemia virus infection.
Tumor necrosis factor alpha (TNF alpha) levels were determined by enzyme-linked immunosorbent assay (ELISA) and by cell culture bioassay in supernatants of lipopolysaccharide-stimulated feline monocyte cultures and in cat serum samples. There was a good correlation between the results obtained by the two methods. From the fact that TNF alpha was neutralized quantitatively by antibodies to human TNF alpha in feline monocyte supernatants and in feline sera, it was concluded that feline TNF alpha immunologically cross-reacts with human TNF alpha and that the human TNF alpha ELISA can be used to quantitate feline TNF alpha. During the first 6 months after experimental feline immunodeficiency virus (FIV) infection no differences in serum TNF alpha values were observed between infected and non-infected cats. TNF alpha levels increased significantly after primary vaccination with a feline leukemia virus (FeLV) vaccine in FIV infected cats over those in the non-infected controls. During secondary immune response TNF alpha levels rose transiently for a period of a few days in both the FIV positive and the FIV negative cats. After FeLV challenge, TNF alpha levels increased in all animals challenged with virulent FeLV for a period of 3 weeks. This period corresponded to the time necessary to develop persistent FeLV viremia in the control cats. It was concluded from these experiments that in the asymptomatic phase of FIV infection no increased levels of TNF alpha are present, similar to the situation in asymptomatic HIV infected humans. Activation of monocytes/macrophages in FIV infected cats by stimuli such as vaccination or FeLV challenge readily leads to increased levels of TNF alpha.
Use of two virustatica (AZT, PMEA) in the treatment of FIV and of FeLV seropositive cats with clinical symptoms.
In the present study the therapeutic efficacy and the side effects of two antiretroviral compounds used in human acquired immunodeficiency syndrome (AIDS) research, 3'-azido-2',3'-dideoxythymidine (AZT, zidovudine, Retrovir) and 9-(2-phosphonylmethoxyethyl)adenine (PMEA), were investigated in the treatment of cats naturally infected with feline immunodeficiency virus (FIV) and cats naturally infected with feline leukemia virus (FeLV). AZT was administered subcutaneously at a dose of 5 mg kg-1 body weight every 12 h and PMEA was administered subcutaneously at a dose of 2.5 mg kg-1 body weight every 12 h during a 3 week hospitalization. The therapeutic efficacy of both compounds was investigated. There was a stronger potency of PMEA than of AZT on the regression of stomatitis in FIV and in FeLV infected cats. In addition, in FIV infection PMEA had a stronger effect on the improvement of the general clinical status. Both antiretroviral compounds were potent agents to improve the immunologic status of FIV infected cats by raising the CD4/CD8 ratio. In FeLV infection PMEA and AZT appeared to reduce antigenemia. The hematological side effects caused by PMEA were severe and stronger than those of AZT. Therefore the advantage of PMEA in clinical and immunologic improvement was diminished by the hematologic disorders, which do not allow long term treatment with this drug in the dose used.
Immunization-induced decrease of the CD4+:CD8+ ratio in cats experimentally infected with feline immunodeficiency virus.
In a previous experiment a group of 15 specified pathogen free (SPF) cats were experimentally infected with a Swiss isolate of feline immunodeficiency virus (FIV). A group of 15 SPF cats served as FIV negative controls. Nine cats of each group were vaccinated with a recombinant feline leukemia virus (FeLV) vaccine, six cats in each group with a placebo vaccine. All vaccinated cats developed high antibody titers to FeLV and were protected against subsequent FeLV challenge infection. In both control groups five of six cats became persistently infected with FeLV. Unexpectedly, the primary immune response to the vaccine antigen was significantly higher in the FIV positive group than in the FIV negative. The secondary response was stronger in the FIV negative cats. The goal of the present investigation was to further study the immune response in these 30 cats. They were immunized twice with the synthetic peptide L-tyrosine-L-glutamic acid-poly(DL-alanine)-poly(L-lysine) (TGAL) 21 days apart. Blood samples were collected on four occasions during the immunization process. They were tested for antibodies to TGAL, complete blood cell counts and CD4+, CD8+ and pan-T-lymphocyte counts. The following observations were made: (1) in contrast to the FeLV vaccine experiment, the primary immune response to TGAL was not significantly stronger in the FIV positive cats when tested by enzyme-linked immunosorbent assay (2). The absolute size of the CD4+ lymphocyte population was distinctly smaller in the FIV positive than in the FIV negative cats. The lowest CD4+ values were found in the dually FIV/FeLV infected cats. (3) A population of CD8+ lymphocytes was identified that was characterized by a distinctly weaker fluorescence. The size of this population increased in FIV positive and decreased in FIV negative cats during the TGAL immunization experiment. (4) The CD4+:CD8+ ratio increased in FIV negative cats during TGAL immunization from 1.9 to 2.3. In contrast, in FIV positive animals the CD4+:CD8+ ratio decreased significantly from 1.9 to 1.3 during the same period. From these and earlier data it was concluded that in short-term FIV infection the immune response to T-cell dependent antigens may be increased over that of the controls. Immune suppression develops gradually with duration of the infection. The significant drop of the CD4+:CD8+ ratio over a 5 week immunization period suggests that antigenic stimulation may accelerate the development of immune suppression in FIV positive cats. If this is a general feature, FIV infection may provide a particularly interesting model for studying the pathogenesis of AIDS.
Retrovirus infections in non-domestic felids: serological studies and attempts to isolate a lentivirus.
An African lioness from the Zoo of Zurich had to be euthanized because of an inoperable tumor. The serum tested negative for feline leukemia virus (FeLV) p27 antigen by enzyme-linked immunosorbent assay (ELISA) but was strongly positive for feline immunodeficiency virus (FIV) antibodies by ELISA and Western blot. When her only offspring and mate were tested for FIV, high antibody titers to FIV were also found in their serum. Lymphocytes were prepared from these two lions on different occasions and co-cultivated with specific pathogen free (SPF) cat lymphocytes in the presence of concanavalin A and recombinant human interleukin-2 (IL-2) for 6 weeks. The cell culture supernatants tested negative for Mg(2+)-dependent reverse transcriptase and FIV p24 by a double antibody sandwich ELISA throughout the culture period. Whole blood and buffy coat cells collected from these two lions were transmitted by intraperitoneal injection into two SPF cats. The two cats did not seroconvert for a period of 11 months nor could reverse transcriptase activity and FIV p24 antigen be demonstrated in the supernatant of several lymphocyte cultures. To determine the importance of lentivirus infections in zoo-kept wild felids, 124 serum samples were obtained from African lions, Indian and Siberian tigers, snow leopards, panthers, cheetahs and other wild cats from nine European zoos. In addition, serum samples collected from 12 Asiatic lions originating from Gir forest in the Indian State of Gujarat were included in this study. The sera were tested for antibodies to FIV, FeLV and feline syncytium-forming virus (FeSFV) by ELISA and Western blot using the respective viruses after gradient purification. In addition, some of the sera were also tested for antibodies to equine infectious anemia virus (EIAV) and Visna-Maedi virus (VMV). Antibodies to FIV were found in 30/53 (57%) of African lions, one of 18 tigers and one of four panthers. All other sera including those collected from the 12 Asiatic lions were negative for FIV antibodies. Some of the FIV positive lion sera had high antibody titers producing strong bands on Western blot strips even in dilutions of >> 1:1000. The Western blot pattern of the lion sera differed from that of domestic cats in that primarily p24 and to a lesser degree p17 was recognized. Antibodies to FeSFV were found in 14 animals (seven with strong, seven with intermediate, reaction). No correlation was found between FIV and FeSFV infection. Antibodies to FeLV were found in two cheetahs which later turned out to have been vaccinated with Leukocell, a FeLV vaccine.(ABSTRACT TRUNCATED AT 400 WORDS)
Prevalence of feline leukemia virus and antibodies to feline immunodeficiency virus in cats in Norway.
Serum samples from 224 Norwegian cats were analyzed for the presence of feline leukemia virus (FeLV) p27 common core antigen, and for antibodies to feline immunodeficiency virus (FIV). Ninety specimens originated from the serum bank at the central referral clinic at the Norwegian College of Veterinary Medicine, which had been collected during the years 1983-1989; 67 sera were submitted from veterinarian practitioners; while 67 sera originated from cats presented for euthanasia. The cats were classified into one "healthy" and one "sick" group. Only 2.2% of sick cats and 1.2% of healthy cats showed FeLV antigenemia, a finding which is lower than which has been reported from many other countries. The prevalence of FIV antibodies was 10.1% in sick cats and 5.9% in healthy cats. Antibodies to FIV was most prevalent in male cats (14.7%) than in female cats (2.1%), and more prevalent among domestic cats (12.0%) compared to pedigree cats (2.4%). Antibodies to FIV in the cats demonstrated increasing prevalence with increasing age. It may be concluded that FeLV causes minor problems in Norwegian cats, while FIV is present in a similar prevalence to what is reported from other countries.
[Clinico-chemical blood parameters in foals in the first two months of life].
Eighteen healthy foals were studied from birth until 2 months of age. Blood samples were obtained at the following times: presuckle, 30 hours, 1, 3, 5, 7 and 9 weeks of age. Changes in serum P, Mg, Ca, Na, K, Cl, iron, AP, ASAT, ALAT, GGT, GLDH, CK, lipase, urea, creatinine, cholesterol, triglyceride, uric acid, protein and fibrinogen and in plasma total solids were examined and the values compared to reference values of adult horses. There were characteristic age related changes in several parameters. Single measurements should be interpreted cautiously to allow for individual variations.
[Spontaneous bacterial peritonitis in a patient with decompensated alcoholic cirrhosis of the liver].
Spontaneous bacterial peritonitis (SBP) constitute a rare complication of cirrhosis of the liver with a high mortality rate. Unspecific sings of decompensation like increasing elaboration of ascites or the development of hepatic encephalopathy are often prevailing. Fever and peripheral blood leukocytosis as well as abdominal sings of peritonitis are not rarely completely missing. There is a strong suspicion of SBP with an ascitic polymorphonuclear count of greater than 500/mm3. Determination of lactate and pH in the ascitic fluid can increase diagnostic sensitivity and specificity if the leukocyte count is dubious. Gram negative rods predominate with over 70% whereas anaerobic organisms appear to be very rare. A 49 year old patient is presented and diagnostic and therapeutic strategies reviewed. In view of the high mortality every clinical deterioration of patients with cirrhosis should alert the physician of the presence of SBP.
Use of western blot and radioimmunoprecipitation for diagnosis of feline leukemia and feline immunodeficiency virus infections.
The uses and limitations of the western blot (WB) and radioimmunoprecipitation assay (RIPA) techniques for study of feline immunodeficiency virus (FIV) and FeLV were evaluated. Western blot analysis was used to detect antigenic relatedness between the 2 lentiviruses. Using a rabbit serum directed against p26 of the equine infectious anemia virus (EIAV) and anti-EIAV horse serum obtained from an infected horse, cross-reactivity with p24 of FIV was revealed. Cat sera obtained late after experimentally induced FIV infection recognized p26 of EIAV, which indicates reciprocal cross-reactivity. For RIPA, FIV was metabolically labeled, and virus-coded proteins were identified, using immunoprecipitation. Polypeptides with apparent molecular mass of about 15, 24, 43, 50, 120, and 160 kilodaltons were detected. An additional polypeptide of 10 kilodaltons was found only by use of WB analysis.
Vaccination of cats experimentally infected with feline immunodeficiency virus, using a recombinant feline leukemia virus vaccine.
A group of 15 cats experimentally infected with a Swiss isolate of feline immunodeficiency virus (FIV) and a group of 15 FIV-negative control cats were inoculated with an FeLV vaccine containing recombinant FeLV-envelope. High ELISA antibody titer developed after vaccination in FIV-positive and FIV-negative cats. Vaccinated and nonvaccinated controls were later challenge exposed by intraperitoneal administration of virulent FeLV subtype A (Glasgow). Although 12 of 12 nonvaccinated controls became infected with FeLV (10 persistently, 2 transiently), only 1 of 18 vaccinated (9 FIV positive, 9 FIV negative) cats had persistent and 2 of 18 had transient viremia. From these data and other observations, 2 conclusions were drawn: In the early phase of FIV infection, the immune system is not depressed appreciably, and therefore, cats may be successfully immunized; a recombinant FeLV vaccine was efficacious in protecting cats against intraperitoneal challenge exposure with FeLV.
HP 0.35, a cephalosporin degradation product is a specific inhibitor of lentiviral RNAses H.
Penicillins, cephalosporins and other betalactam antibiotics are widely used antibacterial drugs. Recently it was found that some of them also have effects on proliferating eukaryotic cells (Neftel, K.A. and Hübscher, U. (1987) Antimicrob. Agents Chemother. 31, 1657-1661), and one such effect was shown to be the inhibition of DNA polymerase alpha (Huynh Do,U., Neftel, K.A., Spadari, S. and Hübscher, U. (1987) Nucl. Acids Res. 15, 10495-10506). The data suggested that degradation products of betalactam antibiotics were responsible for the inhibitory effect on DNA polymerase alpha. There is some confirmation at the structural level, since we found that penicillin binding proteins, the natural target of the cephalosporins, share amino-acid homologies to DNA polymerases and also to reverse transcriptase from HIV1 (Hafkemeyer, P., Neftel, K.A. and Hübscher, U. Meth. Find. Exp. Clin. Pharmacol. 12, 43-46, 1990). We have purified and determined the structure of one product from the cephalosporin Ceftazidim and found one molecule (HP 0.35) that did not interfere with eukaryotic cell proliferation but rather had a specific inhibitory effect on the RNase H activity of human immunodeficiency virus 1 (HIV1) and feline immunodeficiency virus (FIV) reverse transcriptases, while the DNA polymerising activity of these enzymes was not affected. RNases H from HeLa cells, calf thymus and Escherichia coli on the other hand were much less affected by HP 0.35. The inhibitory concentration of 50% (IC50) was more than 10 times lower compared to those of all cellular RNases H. We therefore tested the effect of HP 0.35 on in vitro lentivirus infection as exemplified by FIV-infection of CD(4+)-cat lymphocytes in cell culture. Under conditions where cell proliferation was absolutely unaffected, HP 0.35 was able to inhibit FIV-infection in CD(4+)-cat lymphocytes. Moreover, preincubation of these lymphocytes with HP 0.35 rendered the cells completely unsusceptible to FIV-infection. These data suggest that a degradation product of a clinically used betalactam antibiotic might represent an effective inhibitor class for lentiviral RNase H.
Selenium and vitamin E in blood sera of cows from farms with increased incidence of disease.
Selenium and vitamin E concentrations were measured in the blood sera of 287 cows in 91 herds. Herds with chronic mastitis and muscle diseases had significantly lower selenium concentrations (10.4 +/- 5.26 and 11.7 +/- 8.63 micrograms/litre serum, respectively) than healthy control herds (17.7 +/- 15.00 micrograms/litre). The selenium concentrations in herds with low fertility did not differ from control herds. The vitamin E concentrations were similar in all the herds. Supplementation of the mineral mixture with 20 mg sodium selenite and 2 g vitamin E/kg on 15 farms with a deficient selenium supply increased the selenium concentrations in the blood within two months in 12 of the herds. The vitamin E concentrations were hardly influenced by the supplementation.
Identification of conserved and variable regions in the envelope glycoprotein sequences of two feline immunodeficiency viruses isolated in Zurich, Switzerland.
The nucleotide sequences of the envelope (env) coding regions of two strains of the feline immunodeficiency virus isolated in Zurich, Switzerland (FIVZ1, FIVZ2) have been analysed. In addition, the complete sequence of the FIVZ1 isolate has been determined. Comparisons have been made with the previously published sequences of three North American isolates (PPR and the Petaluma strains FIV34TF10 and FIV14). The isolate FIVZ1 was very similar to the Petaluma strains of FIV and may represent a clonal derivative acquired by 'contamination'. Overall there are between 2.6% and 15.1% amino acid changes in the env gene products of the five isolates. Of the Zurich isolates, FIVZ2 exhibited the greatest divergence to the other viruses and based on its genotype, phenotype and origins probably represents a new isolate of FIV. Possibly the viruses diverged only recently from a common ancestor. Some 31 of the 33 cysteine residues and 17 of the 21 potential N-linked glycosylation sites of the FIV34TF10 env gene product were conserved among all five isolates. The open reading frame 3 (ORF3, or D) which overlaps the env gene (but is encoded in a different frame) has an ATG codon downstream of a potential splice acceptor site in all five isolates, supporting the view that it encodes a viral gene product. In ORF3 of FIVZ1 a stop codon was located 16 amino acids upstream of the stop codon of ORF3 of the other isolates. The ORF4 (or G) of isolate FIVZ2, thought to be the second coding exon of an FIV rev-like gene, contained a nucleotide deletion in amino acid 45 of ORF4, resulting in a--1 frameshift at this position. Comparison of the LTR sequences of the five isolates identified conserved promoter/enhancer elements. A potential stem-loop structure was identified in the R region of the LTRs of all the isolates, despite the heterogeneity of nucleotide sequences in that region. Such structures (TAR) are present in analogous regions of other lentiviruses and are responsible for tat-mediated trans-activation.
[Diagnostic value of sialochemistry. Results of an albumin study].
In 196 patients suffering from disease of the cephalic salivary glands and in 57 healthy controls the concentration of albumin in parotid and submandibular saliva was measured by the immunodiffusion technique. To summarize the results, albumin seems to be a reliable marker of inflammations of the salivary glands even in the early stages and is therefore recommended for the primary diagnostic and the follow-up of acute and chronic inflammatory conditions of the major cephalic salivary glands. In combination with the evaluation of the flow rate, other sialochemic parameters like phosphohexose isomerase and kallikrein activity, B-mode sonography and fine needle aspiration biopsy the frequency of x-ray examination of the radiosensitive salivary glands may be reduced.
[Endoscopic B-mode ultrasonography in the diagnosis of head-and-neck tumors].
Examination of 40 patients with malignant laryngeal and/or pharyngeal tumours shows the possibility of utilising endoscopic B-mode-sonography in ENT. The currently available devices enhance diagnostic certainty of the established radiographic methods only in cases involving oropharyngeal or certain recurrent tumours and blood vessel infiltration. The pros and cons of endoscopic ultrasonography as well as case studies are discussed.
Progressive immune dysfunction in cats experimentally infected with feline immunodeficiency virus.
Within 6 months of infection with the Petaluma isolate of feline immunodeficiency virus, specific-pathogen-free domestic cats exhibited a decrease in the percentage and number of circulating CD4+ lymphocytes and in the CD4+/CD8+ T-cell ratio, along with a marginally significant depression of pokeweed mitogen-induced lymphocyte proliferation in vitro. There was no loss of responsiveness to concanavalin A during this stage, and the cats were capable of mounting a satisfactory antibody response to a T-dependent, synthetic polypeptide immunogen. The pokeweed mitogen response deficit became clearly demonstrable by 11 to 12 months postinfection. A decline in the lymphocyte proliferative response to concanavalin A and a diminished ability to mount an in vivo antibody response to the T-dependent immunogen evolved by 25 to 44 months postinfection. Virus infection did not affect the ability of cats to mount an antibody response to a T-independent synthetic polypeptide immunogen. These data indicate that feline immunodeficiency virus produces a slowly progressive deterioration of T-cell function but does not affect the ability of B cells to recognize and respond to a T-independent antigenic stimulus.
Ototoxicity of vancomycin: an experimental study in guinea pigs.
Vancomycin (V)-a polypeptide antibiotic-is the drug of choice in severe infections with multiresistant staphylococcus spp. Due to the multimorbidity of the patients receiving V, the interactions with known ototoxic agents and the lack of an audiometric documentation, earlier clinical reports of V-associated hearing loss must be carefully interpreted. To elucidate this topic, a controlled animal study considering nonspecific effects was performed. 40 albino guinea pigs with positive Preyer's reflex and normal hearing were used. The hearing function was controlled by means of brainstem auditory evoked potentials before and after intraperitoneal administration of V (75, 150, 300 mg/kg b.w. for 11-17 days), gentamicin (60 mg/kg b.w. for 26 days), neomycin (100 mg/kg b.w. for 17 days) and sodium chloride solution. The cochleas were dissected for hair cell counting by the surface preparation technique. Additionally, the serum concentrations of V were measured. As a result there was no significant difference in hearing loss between the controls and the animals treated with 75 and 150 mg V; toxic doses of 300 mg V led to a certain threshold elevation after click stimuli, but not after trapezoid stimuli of 1, 4 and 8 kHz. On the other hand, the aminoglycosides as a positive control caused an obvious hearing loss after click and trapezoid stimuli, which was correlated to the amount of outer hair cell loss. Renal damage could be excluded by histological examination of the kidneys and estimation of creatinine in serum. Therefore, treatment with moderate doses of V comparable to clinical conditions does not exhibit a specific ototoxicity as compared to the aminoglycosides.(ABSTRACT TRUNCATED AT 250 WORDS)
Comparative determination of selenium in the serum of various animal species and humans by means of electrothermal atomic absorption spectrometry.
It was the goal of this paper to establish total selenium reference values for Switzerland in different animal species and in humans. To this purpose, a flameless atomic absorption method with deuterium background compensation utilizing a graphite furnace atomization system with a pyrolytic platform inside and palladium solution as matrix modifier was developed for the measurement of selenium in serum. The method was characterized by rapid performability, small sample requirement, acceptable detection limit (0.04 mumol/L) and precision and a linear range of measurement up to 4 mumol/L. The method proved to be suited for routine application in species with low selenium concentrations, such as cattle. It may be useful wherever small amounts of serum have to be analysed, e.g. in neonatology. Reference values of serum selenium concentration were determined in healthy subjects of various species, including man. They were as follows (min = 5%, max = 95% percentile values in mumol/L): humans (20-60 years): 0.78-1.48, humans (60-100 years): 0.61-1.73, horse: 0.36-1.68, dog: 1.90-4.31, cat: 3.60-10.09, cattle greater than 9 months: 0.10-0.82, calves 3-9 months: 0.19-0.65, sheep: 0.09-0.54, goat: 0.14-1.42, chicken: 1.68-4.28 and pig: 1.97-3.32. From the data it was concluded (i) that the serum selenium concentration depended on the amount of selenium in the food, and on the type of digestive tract; (ii) that carnivors had the highest and ruminants the lowest selenium values; and (iii) that up to 20% of the geriatric donors had inadequate selenium levels.