Correlation between antifungal susceptibility testing ofCandida isolates from patients with HIV infection and clinical results after treatment with fluconazole

Summary In an open-label controlled study 23 HIV-infected patients (CDC IV A–E) with documented oropharyngeal candidosis were treated with 100 mg fluconazole orally over 5 days (53 episodes; 1–6 treatments/patient). Efficacy data were compared with a control group of 21 patients who received treatment for 10–21 days with 100 mg fluconazole for candidosis. Candida isolates were repeatedly recovered from patients before and after treatment with fluconazole and antifungal susceptibility testing (microbroth-dilution) was done. Inoculum size, medium pH, incubation time and temperature were standardized. Up to 85% of patients responded to therapy clinically and mycologically.Candida albicans was the most important yeast (86%) isolated from cultures of oral washings. In 90% ofC. albicans isolates MIC to fluconazole were low (1.56 mg/l). Primary resistance to fluconazole was not seen, but secondary resistance occurred in two cases clinically andin vitro (MIC25 mg/l). Short treatment for 5 days was as successful as for 10 to 21 days without leading to significantly more recurrences of oral candidosis in these patients. Selection ofCandida spp. other thanC. albicans (e. g.Candida krusei, Torulopsis glabrata) under repeated fluconazole treatment occurred rarely. One patient developed clinical signs of chronic recurrent candidiasis, where onlyC. krusei could be cultured repeatedly.

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Korrelation zwischen MHK-Bestimmung bei Candida-Isolaten von Patienten mit HIV-Infektion und Therapieverlauf nach Behandlung mit Fluconazol

Zusammenfassung In einer offenen kontrollierten Studie wurde bei 23 HIV-infizierten Patienten (CDC IV A–E) eine dokumentierte oropharyngeale Candidose mit 100 mg Fluconazol oral über 5 Tage behandelt (53 Episoden; 1–6 Episoden/Patient). Die Ergebnisse wurden mit einer Kontrollgruppe verglichen, in der 21 HIV-infizierte Patienten über 10–21 Tage mit 100 mg Fluconazol behandelt wurden. Von den Patienten wurden vor und nach jeder Therapie mit Fluconazol Candida-Isolate gewonnen, differenziert und einer Resistenztestung unterzogen (Mikro-Dilutionstechnik). Für die Resistenztestung wurden Inokulumgröße, pH des Mediums, Inkubationszeit und -temperatur standardisiert. Bis zu 85% der Patienten zeigten klinisch und mykologisch eine Heilung/Besserung.Candida albicans war der am häufigsten isolierte Hefepilz (86%) aus sämtlichen Proben, die mit Mundspülungen gewonnen wurden. 90% allerC. albicans-Isolate warenin vitro Fluconazol-empfindlich (MHK 1,56 mg/l). Eine Primärresistenz gegenüber Fluconazol wurde nicht beobachtet, aber in zwei Fällen trat sowohl klinisch als auchin vitro eine Fluconazol-Resistenz gegenüberC. albicans auf (MHK 25 mg/l). Eine Behandlung der Candidose mit 100 mg Fluconazol über 5 Tage führte zu einer vergleichbaren Heilungs-/Besserungsrate wie über 10–21 Tage, ohne daß die Rezidivrate wesentlich erhöht war. Eine therapiebedingte Selektion von Nicht-albicans-Arten(Candida krusei, Torulopsis glabrata) nach Fluconazol-Behandlung wurde selten beobachtet. Allerdings bestand bei einem Patienten der (klinische) Verdacht auf eine durchC. krusei unterhaltende orale Candidose, da nurC. krusei wiederholt nachgewiesen werden konnte.

     

Der Wert der Duplexsonographie nach der orthotopen Lebertransplantation Erfahrungen an 44 Patienten

Zusammenfassung In einer prospektiven Untersuchung wurde an 44 Patienten (33 Männer, 11 Frauen) riach einer elektiven orthotopen Lebertransplantation die Duplexsonographie 196ma1 angewendet. Ziel der Untersuchung war es festzustellen, inwieweit and in welchem zeitlichen Zusammenhang sich die Parameter pulsatiler FluBindex (PFI) und Dämpfungsindex (DI) bei Komplikationen wie Abstoung und Cholangitis verändern. Der Duplex-Doppler-Ultraschall wurde im Mittel 5mal pro Patient durchgeführt. Die letzte Sonographie erfolgte am Entlassungstag. Bei den Messungen fiber der Leberarterie, der Pfortader und den Lebervenen wurden jeweils der PFI und der DI bestimmt. Die Ergebnisse der Indexuntersuchungen wurden mit dem klinischen Verlauf (Abstoung, Cholangitis) sowie mit den Resultaten von 88 Biopsiepräparaten der Lebertransplantate in Beziehung gesetzt. Bezüglich einer histologisch gesicherten AbstoBung ergab der PFI (über der Leberarterie gemessen) eine Sensitivität von 69,4 % and eine Spezifitdt von 72,2 %, der DI (über den Lebervenen gemessen) eine Sensitivität von 89,4% und eine Spezifitdt von 89,1 %. Wenn auch die einfach und beliebig oft anwendbare Untersuchungstechnik die bioptische Sicherung einer Rejektion nicht ersetzen kann, so ist sie in hohem Mae in der Lage, Indikationen zur Biopsie frühzeitig stellen zu lassen und ein Ansprechen auf eine Therapie kurzfristig und mit holier Genauigkeit anzuzeigen.

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Duplex sonography after orthotopic liver transplantation: findings in 44 patients

In a prospective study, 44 patients (11 women, 33 men) who had received orthotopic liver transplants underwent a total of 196 consecutive duplex Doppler ultrasound examinations. The aim of the study was to evaluate the correlation between the pulsatile flow index (PFI) and the damping index (DI) as far as complications as rejection or cholangitis were concerned. The patients were examined five times each on average. The PFI and DI were measured in the hepatic artery, the portal vein and the hepatic veins. The findings were compared with the clinical course (cholangitis, rejection) and the histomorphological diagnosis as determined in biopsy specimens. In biopsy-proven rejection episodes, the sensitivity of the PFI in the hepatic artery was 69.4%, the specificity 72.2%. The sensitivity of the DI in the hepatic vein was 89.4%, the specificity 89.1 %. Combining the two, specificity was more than 90%. PFI and DI in the portal vein bore no apparent relation to clinical course or histomorphological diagnosis. We found duplex Doppler ultrasound extremely beneficial in determining the timing and indication for liver biopsy. In addition, this simple examination, which can be performed as often as desired, accurately shows the transplanted liver's response to measures taken to counter rejection.

     

Response of multicellular tumor spheroids to liposomes containing TNF-α

Summary This publication describes a new model to investigate the influence of tumor necrosis factor- (TNF-) on a three-dimensional glial cell aggregate under defined, standardized, reproducible conditions using the glioma cell line A 172.The cells are initially grown as normal monolayer culture until they reach a cell density of up to 1×10⁶. Subsequently they are grown as spheroids by the liquid overlay technique. Spheroids grown in this way were divided into ten groups of more than 50 cell aggregates. Three groups were coincubated with free TNF- in increasing dosages (100 ng/ml, 200 ng/ml and 1000 ng/ml); three groups were incubated with empty liposomes (0.2 mg/ml, 0.4 mg/ml and 2 mg/ml); three groups received liposomes which had been loaded with TNF-, and one group, which received no treatment, served as control.The diameter of the spheroids ranged from 80 m to 350 m. There was no significant difference in growth between the 3 groups treated with free TNF-. Comparing spheroids treated with TNF- with those which had been coincubated with empty liposomes, there was a significant difference (p<0.001) in growth, which correlated with the amount of liposomes. Similarly, free TNF- had a significantly (P<0.001) stronger growth-inhibiting effect as compared to liposomes loaded with TNF-. Comparing the groups treated with liposomes only to those treated with liposomes loaded with TNF-, the latter exhibited a more marked (although not significantly) growth-inhibiting effect.The preliminary conclusion is that the major growth-inhibiting effect seems to be mediated by the liposomes. This phenomenon is in agreement with results obtained in monolayer cultures.     

The Use of Intracerebral Microdialysis to Determine Changes in Blood-Brain Barrier Transport Characteristics

The aim of this study was to determine whether changes in the transport of drugs into the brain could be determined by in vivo intracerebral microdialysis. Atenolol was used as a model drug to determine blood-brain barrier (BBB) transport characteristics. In rats, unilateral opening of the blood-brain barrier was achieved by infusion of hyperosmolar mannitol (25%, w/v) into the left internal carotid artery. BBB transport, expressed as the ratio of the area under the curve (AUC) of atenolol in brain extracellular fluid over plasma, was three times higher for the mannitol treated hemisphere as compared with the contralateral brain or after infusion of saline, being (mean ± SEM) 0.094 ± 0.024 (n = 16), 0.029 ± 0.007 (n = 12) and 0.030 ± 0.009 (n = 12) respectively. Further evaluation of the data indicated that for experiments performed in the morning the mannitol infusion had little effect on the extent of transport of atenolol into the brain, while in the afternoon BBB transport was about 10-fold higher than in the contralateral and saline group. The mean afternoon ratios ± SEM were 0.155 ± 0.038 (n = 8), 0.012 ± 0.003 (n = 6) and 0.018 ± 0.006 (n = 6) respectively. It is concluded that intracerebral microdialysis is capable of revealing changes in BBB transport and regional and time-dependent differences in drug levels can be demonstrated with the use of this technique.     

Intraperitoneal Versus Interpleural Morphine or Bupivacaine for Pain after Laparoscopic Cholecystectomy

Background: Opioids can produce peripheral analgesic effects by activation of opioid receptors on sensory nerves. This study was designed (1) to examine a novel route of opioid administration, the intraperitoneal injection; (2) to compare this to interpleural application, and (3) to compare opioid with local anesthetic effects under both conditions.

Methods: At the end of laparoscopic cholecystectomy, 110 patients received the following injections in a double-blind, randomized manner: Group 1 (n = 18) was given intraperitoneal morphine (1 mg in 20 ml saline) and 20 ml intravenous saline. Group 2 (n = 17) received intraperitoneal saline and 1 mg intravenous morphine. Group 3 (n = 15) received 20 ml 0.25% intraperitoneal bupivacaine and intravenous saline. Group 4 (n = 20) received interpleural morphine (1.5 mg in 30 ml saline) and 30 ml intravenous saline. Group 5 (n = 20) received interpleural saline and 1.5 mg intravenous morphine. Group 6 (n = 20) received 30 ml 0.25% interpleural bupivacaine and intravenous saline. Postoperative pain was assessed using a visual analog scale, a numeric rating scale, and the McGill pain questionnaire. Pain localization, supplemental analgesic consumption, vital signs, and side effects were recorded for 24 h.

Results: Neither intraperitoneal nor interpleural morphine produced significant analgesia after laparoscopic cholecystectomy (P > 0.05, Kruskal-Wallis test), whereas interpleural bupivacaine was effective (P < 0.05, Kruskal-Wallis test, up to 6 h postoperatively) but not intraperitoneal bupivacaine (P > 0.05, Kruskal-Wallis test). Shoulder pain was not prevalent in the majority of patients during the first 6 h. By 24 h, about half of the patients complained of shoulder pain, which was rated "low" by about one-third of all patients. No significant side effects occurred.     

Increased intracerebral excitatory amino acids and nitric oxide after hypothermic circulatory arrest

BACKGROUND: Prolonged hypothermic circulatory arrest (HCA) results in neurologic injury, but the mechanism of this injury is unknown. This study was undertaken to measure quantitatively intracerebral excitatory amino acids and citrulline, an equal coproduct of nitric oxide, during HCA. We hypothesized that HCA resulted in higher levels of glutamate, aspartate, glycine, causing increased intracellular calcium, and therefore, nitric oxide and citrulline. METHODS: Ten dogs underwent intracerebral microdialysis and 2 hours of HCA at 18 degrees C. Effluent was analyzed by high performance liquid chromatography with electrochemical detection. Five dogs each were sacrificed at 8 and 20 hours after HCA. Neuronal apoptosis was scored from 0 (no injury) to 100 (severe injury). RESULTS: Time course of HCA was divided into six periods. Peak levels of amino acids in each period were compared with those at baseline. Glutamate, coagonist glycine, and citrulline, an equal coproduct of nitric oxide, increased significantly over baseline during HCA, cardiopulmonary bypass, and 2 to 8 hours after HCA. Aspartate increased significantly during HCA and 8 to 20 hours after HCA. Apoptosis score was 65.56 +/- 5.67 at 8 hours and 30.63 +/- 14.96 at 20 hours after HCA. CONCLUSIONS: Our results provide direct evidence that HCA causes increased intracerebral glutamate and aspartate, along with coagonist glycine. We conclude that HCA causes glutamate excitotoxicity with subsequent nitric oxide production resulting in neurologic injury, which begins during arrest and continues until 20 hours after hypothermic circulation arrest. To provide effective cerebral protection, pharmacologic strategies to reduce glutamate excitotoxicity require intervention beyond the initial ischemic insult.     

Helicobacter pylori products upregulate neutrophil superoxide anion production

Helicobacter pylori-neutrophil interactions may play a pathogenic role in H. pylori-induced gastritis and peptic ulcer disease. To understand these interactions, we explored the effects of H. pylori-derived products on neutrophil chemotaxis and superoxide anion production. H. pylori bacteria were cultured and supernatants fractionated. Neutrophil chemotactic activity was confirmed in the crude supernatants and in one fractionated peak corresponding to a previously described neutrophil chemotactic factor. H. pylori-derived crude supernatant, sonicate and all chromatography-derived peaks failed to directly stimulate neutrophil superoxide anion production. However, after pretreatment with sonicate, neutrophils demonstrated increased superoxide anion production (priming) following subsequent exposure to the secretagogue fmet-leu-phe. These results suggest that H. pylori products may attract neutrophils to the gastric mucosa without initially stimulating superoxide anion production or tissue injury. Oxygen radical-mediated gastric mucosal injury may subsequently result when these primed neutrophils undergo additional stimulation by as yet unidentified factors.     

The influence of Ringer's lactate and gelatin infusion on the internal fluid balance of healthy volunteers measured by a non-invasive conductivity technique

Eight healthy male volunteers received in random order at an interval of 1 week 2 litres of Ringer's lactate or 0.8 litre of gelatin (Gelofusine) over half an hour, after overnight fasting. At the end of the infusion period, blood volume and mean arterial pressure had increased significantly in both groups but the increase in blood volume was more pronounced with the colloid. Extracellular fluid volume increased significantly after Ringer's lactate, while a significant decrease was noticed after gelatin. A small decrease in intracellular fluid volume was noted after infusion of Gelofusine, whereas it did not change after infusion of Ringer's lactate. During the 30 min after infusion, blood volume decreased significantly after both treatments but after the colloid it remained higher than the initial value. During the post-infusion period, no significant changes in either intra- or extracellular volume were seen after either treatments. At the end of the study, urine production was significantly more after the Ringer's lactate. It can be concluded that infusion of 0.8 litre of gelatin results in a larger and longer lasting increase in blood volume than 2 litres of Ringer's lactate, probably due to mobilization of extracellular fluid volume. It also leads to extracellular fluid accumulation. The decrease in blood volume after infusion is caused by increased urine production, since no changes were seen in intra- and extracellular fluid volume during this period.     

Pathomorphology of chronization and outcomes of experimental mycoplasma-viral pneumonia]

Pathomorphology of pneumonia induced by successive contamination of the Syrian hamsters with a pathogenic strain of Mycoplasma pneumoniae and an influenza virus A/PR8/34 as well as after contamination with an influenza virus against the background of a previous administration of the mycoplasma membrane fractions was studied. Mixed Mycoplasma-viral infection results in a tumor-like bronchiolar epithelium proliferation having a morphological similarity with bronchiolo-alveolar or acinar carcinoma, influenza virus infection against the background of a previous administration of protein fractions of mycoplasma membranes resulted in a chronization of pneumonia with development of several types of morphological changes: reversible tumour-like proliferation of the bronchiolo-alveolar epithelium, "inflammatory" pseudotumours and sarcoid-like granulomas.     

Research designs appropriate for service agencies

The intent of research is to inform co-workers in similar environments of new or varied approaches to problems assumed to be shared. However, narratives of reported studies neglect to identify variables necessary for reproducing environmental contigencies. If the proposition is accepted that service agencies occupy unique environments, then single organism research designs such as ABAB and multiple baseline designs may provide more valuable information than currently accepted research methodologies.