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Biomedical subjects

R Köhler

Publications and source records attributed to R Köhler.

At least 19 recordsLinked to original sources
PubMed1992

Chimeric prion protein expression in cultured cells and transgenic mice.

The efficient expression of exogenous prion protein (PrP) molecules in mouse neuroblastoma cells that are chronically infected with murine scrapie prions (ScN2a cells; Butler, D.A., et al., 1988, J. Virol. 62, 1558-1564) and in transgenic mice is described. This technology allows investigation of the PrP molecule for structural regions involved in determining species specificity, as well as ablation experiments designed to address the functionality of particular regions of the PrP molecule. Previous reports demonstrated that the PrP gene specifies the host range for susceptibility of transgenic animals to prions (Scott, M., et al., 1989, Cell 59, 847-857; Prusiner, S.B., et al., 1990, Cell 63, 673-686). Consistent with these results, we showed that Syrian hamster (SHa) PrP is ineligible for efficient conversion to PrPSc in ScN2a cells. By constructing a series of chimeric mouse (Mo)/SHaPrP genes, we developed an epitopically tagged functional variant of the MoPrP gene, which can efficiently form protease-resistant PrP molecules upon expression in ScN2a cells. The presence of a defined epitope for an SHa-specific monoclonal antibody allows the products of this chimeric gene to be discriminated from endogenous MoPrP and creates a useful reagent for exploring structure/function relationships via targeted mutagenesis. In addition, we developed a transgenic mouse expression vector by manipulation of an SHaPrP cosmid clone. This vector permits the efficient expression of foreign PrP genes in the brains of transgenic animals, enabling pathological consequences of in vitro mutagenesis to be studied.

Animals↗
PubMed1991

Exogenous and endogenous contributions to nitrogen fluxes in the digestive tract of pigs fed a casein diet. II. Ileal and faecal digestibilities and absorption of amino acids.

The present work aimed at quantifying nitrogen (N) and amino acid (AA) fluxes in the digestive tract of growing pigs fed a casein diet. In this paper we report on digesta passage at the terminal ileum, on apparent balances at the ileal and faecal levels, and on nutrients appearance in the portal vein. Digesta flow-rate at the terminal ileum was maximum between 6 and 12 h after the meal. About 10% of N and 5% of total AA ingested were recovered within 24 h. AA absorption started 30 min after the meal, and was measurable until 13 to 14 h. The total AA absorbed in 24 h accounted for 128% of the AA ingested. The AA composition of ileal digesta was very different from that of casein, closely resembling that of endogenous proteins. The AA composition of faeces was very close to that of bacterial proteins. The ileal digestibilities of AA, though lower than their faecal values, were very high. This was confirmed by AA absorption balances greater than 100%. These data suggest that casein was almost totally digested by the terminal ileum, and that endogenous AA were substantially reabsorbed. These findings are supported by data on endogenous N recycling (15N), reported in a following paper.

Amino Acids↗
PubMed1991

A rabies-specific human monoclonal antibody that protects mice against lethal rabies.

According to a recommendation from WHO (World Health Organisation) for prevention of a possible rabies infection, active vaccination has to be combined with application of immunoglobulin to get a fast protective effect. At present, preparations of purified human or equine rabies-specific immunoglobulin are used. We have generated a human rabies-specific monoclonal antibody (huMAb) by immortalization of human B-cells with Epstein Barr Virus (EBV), followed by fusion with a mouse myeloma cell. The resulting clone TW-1 secrets an IgG1 lambda huMAb which specifically reacts in ELISA with 5 laboratory rabies virus strains of serotype 1 and DUV3 (Duvenhage, serotype 4). Western Blot analysis revealed fine specificity for the G glycoprotein (gp67) of rabies virus. HuMAb TW-1 neutralizes rabies virus in vitro (RFFIT) as well as in vivo and protects rabies infected mice. Compared to polyclonal human rabies immunoglobulins, huMAb TW-1 is advantageous, because of its defined specificity and the very low amounts of total protein needed for therapeutic effects.

Animals↗
PubMed1990

Escherichia coli-derived envelope protein gD but not gC antigens of herpes simplex virus protect mice against a lethal challenge with HSV-1 and HSV-2.

Immunization studies with HSV-1 and HSV-2 envelope proteins expressed in Escherichia coli were performed. After active immunization of mice with a gD-1 antigen (Leu53-Ala312) expressed as a fusion protein, the animals were protected from a lethal challenge with HSV-1 and HSV-2. In addition, antisera from rabbits immunized with the same gD-1 antigen also conferred passive immunity to mice against a challenge infection with either HSV-1 or HSV-2. In contrast to these successful gD-1 protection experiments, various gC-1 and gC-2 fusion proteins from E. coli failed to induce protective immunity. Moreover, the mice sera from immunized animals were not able to react with the authentic, glycosylated gC-1 and gC-2 envelope proteins, whereas sera raised against authentic gC-1 and gC-2 glycoproteins do recognize the gC fusion proteins from E. coli. These results indicate, that E. coli might represent an ideal system for expressing gD antigens as a possible component of a HSV vaccine, whereas gC antigen cannot be produced in an immunocompetent form in E. coli.

Animals↗
PubMed1990

[A modified Sarafoff suture for closure of the uterine wound in Cesarean section].

Using a retrospective investigation the authors demonstrate the advantage of the modified Sarafoff-suture (a one layer suture of the myometrium) in comparison with the two-layer suture. The results shows a lower rate of febrile courses, less postoperative complications and a shorter hospital stay. The first clinical experiences showed less scar complications in the courses of vaginal delivery in patients with a previous caesarean section. Therefore, the authors advise the modified Sarafoff-suture for wound closure in caesarean section.

Cesarean Section↗
PubMed1989

Blood levels of glucometabolic hormones and urinary saturation with stone forming phases after an oral test meal in male patients with recurrent idiopathic calcium urolithiasis and in healthy controls.

Male patients with recurrent calcium (Ca) urolithiasis (RCU) with idiopathic hypercalciuria (I-HC, n = 12) or normocalciuria (NC, n = 12), and age, sex, and weight-matched controls (C, n = 12) were evaluated before and after a carbohydrate-rich synthetic meal for blood glucose, free fatty acids (FFA), alpha-amino-nitrogen, several glucometabolic hormones and parathyroid hormone (PTH), and urine Ca, phosphate, oxalate, and cyclic adenosine monophosphate (cAMP) levels as well as saturation. Fasting serum Ca was significantly higher and PTH significantly lower in I-HC than in controls, whereas in fasting urine cAMP and phosphate were unchanged. There were only minor differences between fasting blood glucose levels and postprandial glucose tolerance of RCU patients and controls. However, serum insulin was significantly elevated in I-HC versus C, but serum C-peptide, plasma glucagon, and somatostatin levels were comparable in RCU and C. FFA were significantly lower in RCU than C. Postprandial phosphaturia and urinary saturation with Ca-phosphates were significantly higher in RCU versus C, whereas urinary cAMP, pH, and oxalate were similar. We conclude that: (1) in RCU patients some postabsorptive steps in glucose metabolism may be abnormal; (2) those with I-HC have enhanced postprandial Ca and phosphate excretion concomitantly with disordered insulin metabolism; and (3) RCU patients may suffer from a postprandial renal phosphate leak, which may make their urine more lithogenic.

Adult↗
PubMed1988

A sex cord stromal tumour in a woman with XO/XX/XXX-mosaicism.

We describe a postmenopausal woman with 45,XO/46,XX/47,XXX mosaicism who presented with a Sertoli-Leydig cell tumour and a past history of habitual abortion. The cytogenetic finding is discussed in relation to the etiology of the tumour and of the recurrent abortions.

Female↗
PubMed1988

Urinary pyrophosphate in patients with recurrent calcium urolithiasis and in healthy controls: a re-evaluation.

The excretion of inorganic pyrophosphate was studied in daily, fasting and postprandial urine specimens of normocalciuric and hypercalciuric patients with recurrent renal calcium stone disease (40 men and 40 women), and healthy controls (20 men and 20 women). Both populations were subdivided into younger (20 to 40 years old) and older (more than 40 years old) individuals. In general, there was a tendency towards higher urinary pyrophosphate excretion with increasing age (both sexes and all groups studied), and lower excretion in women than in men. The urinary pyrophosphate excretion rate was unchanged in daily and fasting urine specimens of the younger male normocalciuric and idiopathic hypercalciuric stone patients, whereas in the daily and postprandial urine of younger women the median excretion rate was reduced (controls versus normocalciuric plus idiopathic hypercalciuric subjects, 3 versus 1 mumol., p less than 0.05). In contrast, in older men urinary pyrophosphate was reduced in daily specimens (controls versus normocalciuric plus idiopathic hypercalciuric subjects, 55 versus 33 mumol., p less than 0.05) but it was unchanged in fasting urine specimens. In older women no change was detectable in any of the 3 urine portions. Factorization of urinary pyrophosphate for the associated urinary creatinine did not alter these results substantially, and the presence of renal stones did not modify pyrophosphate excretion significantly. Urinary pyrophosphate was correlated significantly with urinary volume, citrate and phosphorus. We conclude that only subclassification of stone patients with respect to sex, age and type of calciuria, and consideration of additional urine portions besides the daily urine may help to uncover states of urinary pyrophosphate deficit. On the basis of the data, we recommend that clinically relevant studies on inhibitory effects of urinary pyrophosphate on the nucleation and growth of crystals and stones should be done preferentially in urine portions with a proved pyrophosphate deficit.

Adult↗
PubMed1987

Fasting gastrinemia and elevated supersaturation with hydroxyapatite of fasting urine--observations in renal calcium stone patients and controls.

We evaluated serum gastrin, acid-base status, variables of mineral metabolism in fasting blood, as well as pH, relative supersaturation of stone forming constituents, and crystalluria in the associated fasting urine, of control subjects (n = 12), and in age- and weight-matched male normocalciuric (n = 12) and hypercalciuric (n = 12) patients with idiopathic recurrent calcium urolithiasis (RCU). In RCU, mineral metabolism and acid-base data are unchanged, whereas mean serum gastrin is only insignificantly higher as compared to controls. Subclassification of all participants into categories with either high-normal or low-normal gastrin reveals that in RCU with low-normal gastrin there is a higher-than-normal urinary pH and significantly elevated supersaturation of urine with hydroxyapatite. Crystalluria and stone analysis support the assumption that the physico-chemical environment accompanied by low gastrin levels predisposes to urinary precipitation of calcium phosphate with subsequent formation of a stone nidus. pH in fasting urine and integrated fasting serum gastrin correlate significantly, suggesting that low fasting serum gastrin in RCU patients may be considered a risk factor for calcium phosphate stone formation.

Acid-Base Equilibrium↗
PubMed1987

Combined antiviral effects of acyclovir or bromovinyldeoxyuridine and human immunoglobulin in herpes simplex virus-infected mice.

Suboptimal dosage regimens of antivirals (acyclovir or bromovinyldeoxyuridine) and human immunoglobulin have been combined in the treatment of hairless mice infected with herpes simplex virus type 1. The aim of this study was to establish how late after infection human immunoglobulin could be given to still be effective and for how long would the protective effect last. The combination of acyclovir or bromovinyldeoxyuridine with passive immunization proved additive or even synergistic depending on the time of immunoglobulin administration and the observation period after infection. When the survival rate of the mice was followed for up to 20 days postinfection, synergistic action seemed to occur with immunoglobulin given as late as 2 or 3 days after infection. When the mice were followed for up to 100 days after infection, however, it turned out that only the immunoglobulin given 4 h after infection led to a synergistic effect with the antivirals. Most of the mice subjected to combined treatment, in contrast to mice treated with the antivirals only, did not develop anti-HSV antibodies. This lack of a specific humoral immune response possibly reflects the rapid inhibition of virus replication early after challenge by the combined treatment, thus preventing the production of a sufficient amount of viral antigen in the body needed for a measurable antibody induction.

Acyclovir↗
PubMed1987

Fasting and post-calcium load serum calcium, parathyroid hormone, calcitonin, in male idiopathic calcium urolithiasis--evidence for a basic disturbance in calcium metabolism.

In male healthy controls (n = 12), male renal calcium stone patients with either normocalciuria (NC; n = 12) or idiopathic hypercalciuria (I-HC; n = 12), all ideally matched for age and body weight, the response of variables of mineral metabolism to a calcium-rich oral test meal was evaluated. In all groups the postprandial urinary cyclic AMP is decreased as compared with fasting urine, indicating that the parathyroid glands are suppressible by oral calcium. However, in I-HC serum total calcium was significantly higher than in controls both basally and in the postprandial period; the associated mid-regional (bioinactive) serum parathyroid hormone was also elevated, but serum parathyroid hormone recognizing the amino terminal (bioactive) region of the molecule was significantly decreased. Also in I-HC, serum alkaline phosphatase is elevated, whereas urinary hydroxyproline in both fasting and postprandial urine is unchanged. The NC group holds an intermediate position between I-HC and controls. It can be concluded that I-HC subjects may suffer from a more basic disturbance of calcium metabolism which may help explain the nature of their hypercalciuria.

Adolescent↗
PubMed1987

[Methodologic aspects of the determination of N-exchange parameters from 15N-tracer studies in swine on the basis of models of N-metabolism. 3. Calculation of protein synthesis and decomposition rate on the basis of lysine flux, ascertained from the amount of 15N in the urine].

The final product method introduced here is based on the use of 15N L-Lysine as tracer amino acid for the determination of the protein synthesis and decomposition quotas of the total body with the flux of lysine. For this purpose it is necessary to complete the N flow and N compartments of the 3-pool model by the values of lysine content. The ascertained values of protein synthesis--8.4 g/d/kg--and protein decomposition--6.9 g/d/kg--agree very well with those determined with 15N glycine as tracer amino acid.

Animals↗
PubMed1987

Effects of acute oral sodium potassium citrate load in healthy males--outlook for treatment of patients with calcium-containing renal stones.

The acute effects of a single (5 g) oral load of sodium potassium citrate (SPC), given together with a liquid test meal, were studied in 6 healthy male volunteers with respect to changes in serum citrate, blood acid base status, urine pH, citrate, calcium and minerals, and oxalate, as well as the calculated relative supersaturation of urine with several stone-forming phases, and the associated crystalluria. It was found that, apart from making the urine more alkaline, SPC induces mild compensated metabolic alkalosis, increases serum and urinary citrate, and reduces fractional urinary calcium excretion, but leaves urinary oxalate and the accompanying crystalluria unchanged. Except for the increase in urinary supersaturation with hydroxyapatite, the supersaturation of other important stone-forming constituents is statistically unchanged. In addition, there are indications that SPC reduces postprandial intestinal calcium absorption without affecting serum parathyroid hormone and 1,25-dihydroxyvitamin D. It is concluded that there is a spectrum of acute effects of oral SPC that may warrant long-term trials on this medication in the metaphylaxis of calcium-containing urinary stones.

Adult↗
PubMed1986

[Determination of N metabolism parameters in an N increase trial in the rat using 15N tracers].

3 groups of 8 Wistar rats each of a live weight of ca. 215 g received graded N supplements. A number of N metabolism parameters were ascertained and discussed. Among other things, the protein synthesis and degradation quotas were ascertained by means of the 15N tracer technique. For the assessment a 3-pool model was used, the content of which was in so far extended as growth and metabolic faecal nitrogen are concretely taken into consideration. A quasi-continuous method of supplementing the tracer was chosen; it was discontinued in approximately the middle of the experiment. Thus the labelling and abatement phases in urine could at the same time be included in the calculation. The protein synthesis quota increases with N intake. It can be concluded from the course of the graph that it gravitates towards a saturation value.

Animals↗