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Biomedical subjects
Publications and source records attributed to C Bühler.
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BACKGROUND: The aim of this study was to determine the efficacy of three viscoelastics of different viscosities in preventing early postoperative complications after trabeculectomy (TE) and phacotrabeculectomy (PT). METHODS: One hundred seventy-one eyes of 149 consecutive patients underwent TE or PT. In all cases, Healon, Healon GV, or Healon 5 was instilled in the anterior chamber at the beginning of surgery and left there at the end. RESULTS: Four hours postoperatively, the mean intraocular pressure (IOP) in all groups was below 13 mmHg (27.4 mmHg preoperatively) and did not rise above 13 mmHg in the first postoperative week. Postoperative IOP spikes did not occur in any group. Healon remained in the anterior chamber up to 4 days after surgery, Healon GV up to 6 days, and Healon 5 up to 9 days. Characteristic early postoperative complications of filtering procedures, such as choroidal detachment, shallow anterior chamber, iridocorneal contact, hyphema, and bleb failure, were not observed in our patients. CONCLUSION: All three viscoelastics reduced early postoperative complications with comparable efficacy and without IOP spikes.
UNLABELLED: Xenogeneic transplantation of porcine islets of Langerhans is regarded as a potential future treatment for diabetes mellitus. Despite considerable biotechnological progress, however, it is still very difficult and often unreliable to isolate sufficient numbers of highly purified, intact islets from the porcine pancreas with good in vitro function. OBJECTIVE: Of this study was to describe an efficient and reliable method to isolate sufficient numbers of highly purified islets of Langerhans with good in vitro function from adult as well as from young hybrid pigs. METHODS: Islets were isolated from the pancreas of young (4-6 months) hybrid pigs and old (2-3 years) retired breeders using Liberase PI and digestion-filtration. Average islet size was detected by dithizone staining of tissue sections prior to isolation; only organs with an average islet size > or = 200 microns were used. Density gradient purification with OptiPrep was performed in a COBE 2991 cell processor. Viability was investigated using fluorescence staining. Perifusion studies were carried out to asses in vitro function of isolated islets. RESULTS: Islets were successfully isolated from young hybrid pigs (3,671 +/- 598 IEQ/g) and old retired breeders (5,182 +/- 545 IEQ/g). After purification islet purity was 92% for retired breeders and 87% for young hybrid pigs. Yield after purification was still not satisfactory: 64% for retired breeders (3,209 +/- 444 IEQ/g) and 44% for young hybrid pigs (1,669 +/- 386 IEQ/g). Viability of isolated islets was 80-95%. Perifusion studies of porcine islets showed sufficient insulin release upon glucose challenge; however, the level of insulin release depended on the density of islets within the perifusion chamber. Low temperature culture (24 degrees C) prior to perifusion studies had no detrimental effect on insulin release. Long-term culture over 11 days was followed by a dramatic loss of islet function. CONCLUSIONS: If xenograft rejection can be overcome and the risk of xenosis can be minimised, sufficient numbers of purified porcine islets with good in vitro function can be isolated to serve as a potential source for islet transplantation in diabetic patients.
Transplantation of isolated pancreatic islets provides an interesting alternative to the present cure for diabetes: insulin injections and pumps. These are characterized by an insufficient glucose haemostasis and in the long run can induce kidney failure, blindness, heart failure, and amputations. Up to now more than 293 allogeneic islet transplantations have been performed in diabetics with chronical kidney failure. Despite some success, no real breakthrough has been yet achieved, though great efforts are being made to improve the various methodological steps on the way to clinical transplantation. The use of animal (xenogeneic) organs could be a solution to overcome the shortage of allogeneic donors. The current experimental and clinical research focuses on the use of pigs as organ donors, which have a number of advantages over the immunologically more compatible primates. This article reports on success and open questions concerning the efforts to isolate porcine islets for future clinical transplantation: the search for a suitable pig breed, the various isolation steps, purification and in vitro culture, transplantation models using-small and large animals, first clinical trials, and immunological reactions against the xenogeneic tissue. In addition, strategies to circumvent tissue rejection and future perspectives are discussed.
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The effects of feeding different amounts of colostrum or only milk replacer and the effects of Long-R3-IGF-I (administered s.c. or orally; 50 microg/[kg BW x d] for 7 d), and of s.c. injected recombinant bovine GH (rbGH; 1 mg/[kg BW x d] for 7 d) on small intestinal mucosal morphology in newborn calves were studied by histomorphometry. Neonatal calves fed colostrum six times exhibited greater (P < .01) villus circumferences, areas, and heights in total small intestine and especially in the duodenum than calves fed only milk replacer. Furthermore, villus circumferences and areas in total small intestine were greater (P < .05) in calves fed colostrum once than in calves fed no colostrum. Villus size in total small intestine was smaller (P < .05) in rbGH-treated than in control calves; jejunum villus circumferences and heights were especially reduced (P < .05). Crypt depths in ileum were greater (P < .05) in rbGH-treated calves. In conclusion, prolonged colostrum supply significantly enhanced small intestinal villus size in neonatal calves. In contrast, Long-R3-IGF-I had no significant influence on small intestinal morphology, and rbGH in supraphysiological amounts even reduced small intestinal mucosal variables after 1 wk of treatment. The study demonstrated enhanced postnatal development of the gastrointestinal tract by prolonged colostrum feeding, but not by Long-R3-IGF-I or GH.
Microscopy is traditionally a tool for determining biological structures. Many recent advances in optical microscopy involves the incorporation of spectroscopy techniques to monitor biochemical states of microscopic structures in living cells and tissues. By minimizing tissue photodamage, two-photon excitation microscopy provides a new opportunity to study the dynamics of biological systems on time scales from nanoseconds to hours. This review will focus on a number of these new methods: two-photon time-lapse microscopy, two-photon photoactivation, two-photon correlated spectroscopy, two-photon single particle tracking and two-photon lifetime microscopy.
To determine the prevalence of bacteremia caused by Bartonella henselae in domestic cats in the region of Freiburg, Germany, we investigated culture of blood from 100 cats from 89 different households over a 12-month period. B. henselae could be isolated from 13% (13 of 100) of these cats. In eight households with two cats each and in one household with three cats, B. henselae bacteremia was found either in all of the animals or in none of the animals. Positive cultures were more likely to be found for female, young (24 months of age or younger) cats than for male or older cats. Identification of the Bartonella isolates was made by colony morphology, by Gram staining, biochemically by RapID ANA II or Rapid ID 32 A systems, and by whole-cell fatty acid analysis. Differentiation between B. henselae and Bartonella quintana was only possible by 16S rRNA sequencing, enterobacterial repetitive intergenic consensus (ERIC)-PCR and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Genomic fingerprinting of the B. henselae isolates by ERIC-PCR yielded two different patterns based on three distinct bands.
In order to select those patients with colorectal liver metastases who may have benefit from an operation, preoperative detection and localization of tumors in the liver is necessary. In this study, the specificity and sensitivity of a CT arterial portography and magnetic resonance imaging with contrast media was compared with the results of operative findings and intraoperative ultrasonography. The combination of both preoperative methods led to an accuracy rate of more than 90%, and all patients who underwent laparotomy were resected or treated with an arterial infusion port system. Negative laparotomies were excluded completely.
Bartonella quintana was grown from a lysis-centrifugation blood culture of an HIV-positive man who presented with lesions of bacillary angiomatosis in skin and bones. The isolate was identified by sodium dodecylsulfate-polyacrylamide gel electrophoresis, 16S rRNA gene sequencing, and amplification of the enterobacterial repetitive intergenic consensus sequences.
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The hypoglycaemic sulphonylurea gliquidone and glibenclamide exerted a partial uncoupling effect on mitochondrial respiration of liver under in vitro conditions using various citrate cycle intermediates as substrates. Besides the uncoupling effect, gliquidone and glibenclamide caused a direct inhibition of ATP-as well as DNP-stimulated oxygen consumption. Both phenomena proved to be dose dependent. Respiratory control ratio decreased progressively with increasing concentrations of sulphonylureas mainly through the inhibition of ADP-stimulated respiration. Basal and DNP-stimulated ATP-ase activity of isolated mitochondria changed similarly to the respiratory parameters. Changes in membrane permeability of mitochondria and the inhibition of substrate uptake further support the assumption of structural and functional alteration of mitochondria by the hypoglycaemic compounds tested.
Alterations in circadian blood pressure rhythm have been described in patients with sleep apnea syndrome or arterial hypertension. Epidemiologic data indicate an association of circadian blood pressure rhythm and obstructive snoring; however, only little is known about effects of apneic phases during sleep on arterial hypertension and their relevance for management of hypertension. Effects of arterial desaturation phases caused by apnea or hypopnea during sleep on arterial blood pressure have been investigated in altogether 16 probands. Thereby, a significant reduction of circadian blood pressure rhythm (p < 0.003) and of the difference between daily and nocturnal diastolic blood pressure values (p < 0.002) was found with increasing apnea index. Our results show that the sleep apnea syndrome or obstructive snoring, respectively, should be integrated more often in diagnostic and therapeutic management of arterial hypertension, especially when the circadian blood pressure rhythm appears to be levelled by ambulatory 24-hour monitoring of blood pressure values.
The involvement of the mitochondrial bound hexokinase in aerobic glycolysis was investigated in two subpopulations of the HT 29 human colon cancer cell line: a poorly differentiated one with high aerobic lactate production (referred as undifferentiated or standard cells), and an enterocyte-like differentiated one with lower lactate production (referred as differentiated or Glc- cells). After mild digitonin treatment, 85% of the total cellular hexokinase activity remained in the particulate fraction in both cell types. In both cases mitochondria appeared to be tightly coupled but the Glc- cells exhibited a significantly higher oxidation rate in the presence of glucose. Electron microscopy of freeze-fractured cells revealed the absence of contacts between the two limiting mitochondrial membranes in the highly glycolytic standard cells, whereas the contacts were present in the Glc- cells. Furthermore, we investigated the functional relationship between bound hexokinase (as hexokinase-porin complex) and the inner compartment of mitochondria isolated from standard and Glc- HT 29 cells. In contrast to the differentiated cells the hexokinase in undifferentiated standard cells was not functionally coupled to the oxidative phosphorylation. This suggests that the high rate of lactate formation in neoplastic cells is not caused by an increase of particulate hexokinase activity but rather by a disregulation of the hexokinase-porin complex caused by the absence of contact sites between the two mitochondrial membranes. In agreement with this interpretation, the hexokinase-porin complex could be completely removed by digitonin treatment in standard HT 29 cells, while this was not possible in mitochondria from Glc- cells.
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