Methods for the determination of the sensitivity of Candida strains to fluconazole: focus of a micromethod and correlation with 2 other techniques

107 Strains of different species of Candida have been tested by three methods of susceptibility to fluconazole. In addition, one reference strain has been used to verify three methods' reliability: one diffusion method on Shadomy's medium and two MIC liquid methods, on tube and on microtiter plate. Method on Shadomy's medium seems less sensitive for yeast susceptibility. Comparison of two liquid method of MIC show discordances on resistant strains (14% for method on tube and 26% for micromethod) By simplicity of execution and reliability in detection of natural resistances, micromethod of MIC seems the most convenient for yeasts determination in vitro to fluconazole.     

Time-of-flight MR angiography of the portal venous system: value compared with other imaging procedures

A simple method for determination of mimosine and 3,4-dihydroxypyridine (3,4-DHP) in plasma and milk was developed. Milk and plasma, with tyrosine as internal standard, were deproteinized using 9% trichloracetic acid and extracted with diethyl ether. Metabolites were separated by isocratic high-performance liquid chromatography, with 0.02 M orthophosphoric acid (pH 2.5) at 0.5 ml/min and a Hypersil ODS microbore column. Mimosine, 3,4-DHP and tyrosine were detected at 275 nm. The recovery of the mimosine added to the plasma samples was 101.6 +/- 2.3% and 103.3 +/- 1.0% for milk samples. 3,4-DHP recovery for plasma samples was 101.2 +/- 0.9% and for milk samples 100.8 +/- 1.4%. The reproducibility of the method was evaluated by analyzing six plasma samples and six goat milk samples. The analyses yielded relative standard deviations of 2.65 and 2.82%, respectively.     

Interaction of the renin-angiotensin system, bradykinin and sympathetic nerves with cholinergic transmission in the rat isolated trachea

1. The present study was undertaken to investigate the interaction of the renin-angiotensin system (RAS), bradykinin and the sympathetic nervous system with cholinergic transmission in the rat airways. Experiments were performed on epithelium-intact and epithelium-denuded preparations of rat isolated trachea which had been incubated with [3H]-choline to incorporate [3H]-acetylcholine into the cholinergic transmitter stores. Tracheal preparations were subjected to electrical field stimulation (trains of 1 ms pulses, 5 Hz, 15 V) and the stimulation-induced (S-I) efflux taken as an index of transmitter acetylcholine release. 2. In both epithelium-intact and epithelium-denuded tracheal preparations, the alpha 2-adrenoceptor agonist UK14304 (0.1 and 1 microM) inhibited the S-I efflux, in a concentration-dependent manner. The inhibition of S-I efflux produced by UK14304 (1 microM) was antagonized by the selective alpha 2-adrenoceptor antagonist idazoxan (0.3 microM). Idazoxan (0.3 microM) alone had no effect on the S-I efflux. 3. Angiotensin II (0.1 and 1 microM) was without effect on the S-I efflux in either epithelium-intact or epithelium-denuded tracheal preparations. When angiotensin-converting enzyme was inhibited by perindoprilat (10 microM), angiotensin II (1 microM) was also without effect on the S-I efflux. Similarly, in the presence of idazoxan (0.3 microM), to block prejunctional alpha 2-adrenoceptors, angiotensin II (0.1 and 1 microM) did not alter the S-I efflux. When added alone, perindoprilat (10 microM) did not alter the S-I efflux. 4. In epithelium-denuded preparations, bradykinin (0.01-1 microM) inhibited the S-I efflux. In epithelium-intact preparations, there was also a tendency for bradykinin (0.1 and 1 microM) to inhibit the S-I efflux but this was not statistically significant. However, when angiotensin-converting enzyme and neutral endopeptidase were inhibited by perindoprilat (10 microM) and phosphoramidon (1 microM), respectively, bradykinin (1 microM) significantly inhibited the S-I efflux in epithelium-intact preparations as well as in epithelium-denuded preparations. The inhibition of the S-I efflux produced by bradykinin, in the combined presence of perindoprilat (10 microM) and phosphoramidon (1 microM), was unaffected by the additional presence of the cyclo-oxygenase inhibitor indomethacin (10 microM) and/or the nitric oxide synthase inhibitor NG-nitro-L-arginine (100 microM), in either epithelium-intact or epithelium-denuded preparations. 5. In conclusion, the findings of the present study suggest that airway parasympathetic nerves are endowed with alpha 2-adrenoceptors which subserve inhibition of transmitter acetylcholine release. Under the present conditions, however, transmitter acetylcholine release is not subject to transneuronal modulation by noradrenaline released from adjacent sympathetic nerves in the airways. Moreover, angiotensin II and perindoprilat do not appear to modulate acetylcholine release from parasympathetic nerves of the airways. In contrast, bradykinin inhibits acetylcholine release from airway parasympathetic nerves but this action of bradykinin is limited by the activity of epithelial angiotensin-converting enzyme and/or neutral endopeptidase. The inhibitory action of bradykinin on cholinergic transmission in the airways does not appear to involve the liberation of prostaglandins or nitric oxide.     

Thrombosis of the deep cerebral veins: CT and MRI findings with pathologic correlation

Deep cerebral vein thrombosis can present with acute, severe neurological symptoms and may be rapidly fatal as in the 20-year-old woman reported here. Although MRI is superior for establishing the diagnosis, CT is usually the first examination performed in the clinical setting. It is therefore important to recognise certain indicators such as extensive bithalamic low density. These and certain other less specific signs are correlated with the MRI and autopsy findings.     

Orbital venous approach to the cavernous sinus: an analysis of the facial and orbital venous system

Carotid-cavernous fistulas are abnormal communications between the internal carotid artery and the cavernous sinus produced by a rupture of the wall of the carotid artery or one of its branches into the sinus. Extradural branches of the internal or external carotid arteries may communicate with the cavernous sinus, producing proptosis, progressive glaucoma, and ocular vascular engorgement. Various approaches to obliterate these fistulas have evolved, many of which carry high morbidity or are precluded by anatomical considerations. Analysis of the venous anatomy of the orbit and face, including human cadaver dissections, reveals a new and safe approach to the cavernous sinus, requiring microsurgical isolation and cannulation of the superior ophthalmic vein through an anterior orbital approach. Selective embolization of a carotid-cavernous fistula can be performed successfully through this route. We present pertinent anatomy and technical considerations and the successful clinical application of these principles. Surgeons familiar with craniofacial anatomy and microvascular techniques can apply these principles and play an active role in the treatment of these complex problems.     

Functional expression of Fas (CD95) in acute myeloid leukemia cells in the context of CD34 and CD38 expression: possible correlation with sensitivity to chemotherapy

Clinical studies of bone marrow transplantation (BMT) suggest that the immune system contributes to the eradication of acute myeloid leukemia (AML). A recent study also showed that the Fas (CD95/APO1) mediates apoptotic signal from cytotoxic T lymphocytes. Sixty-four patients with AML were studied for the expression of Fas in the context of CD34 and CD38 coexpression. The clinical relevance of Fas expression and function on AML was also investigated. Fas was expressed on 2% to 98% of AML cells (2% to 20% in 11 patients, 20% to 50% in 20 patients, 50% to 80% in 24 patients, and 80% to 98% in nine patients). Only 44.4% of patients with AML M1 (French-American-British [FAB] classification) were Fas+ (>/=20% of leukemia cells expressed Fas), whereas 89.1% of patients with AML M2, M3, M4, M5 were Fas+ (P < .01). Among 43 CD34+ patients (>/=20% leukemia cells were CD34+), 34 were Fas+, and 19 of 21 CD34- patients were Fas+ (P = NS). Thirteen cases were studied for their expression of Fas in the context of CD34 and CD38 using three-color analysis. Fas is expressed at a high level in the gated CD34+CD38+/- and CD34+CD38+ population. In 10 AML samples, Fas was expressed at a higher level in CD34+/CD38+ population than in CD34+/CD38+/- or CD34- cell populations. Fas-induced apoptosis by anti-Fas monoclonal antibody (MoAb) was determined by morphologic features and colorimetric DNA fragmentation assay. Induction of apoptosis was found in 14 of 24 cases. However, no statistically significant correlation was observed between Fas expression and induction of apoptosis. Leukemia colony-forming unit assays suggested that in some cases, Fas-induced apoptosis occurred in the clonogenic cell populations. Parameters such as laboratory and clinical data at initial diagnosis were correlated with Fas expression and only response to initial induction chemotherapy showed significant correlation with Fas expression (P < .05). We conclude that the majority of AML cells exhibit variable expression of Fas, and apoptosis could be induced by anti-Fas MoAb in some cases. Our results suggest the Fas-mediated apoptosis may be clinically relevant, whereas the issue of clonogenic leukemia cells and Fas expression needs further studies.     

A comparison of the effects of bradykinin, 5-hydroxytryptamine and histamine on the hepatic arterial and portal venous vascular beds of the dog: histamine H1 and H2-receptor populations

1 The hepatic arterial and hepatic portal venous vascular beds of anaesthetized dogs were separately perfused in different experiments.2 From measurements of perfusion pressures and blood flows in the two series of experiments, hepatic arterial vascular resistance (HAVR) and hepatic portal venous vascular resistance (HPVR) respectively were calculated.3 Bradykinin, 5-hydroxytryptamine (5-HT) and histamine were injected intra-arterially and intra-portally and dose-response curves constructed from these data.4 Bradykinin injected intra-arterially caused dose-dependent hepatic arterial vasodilatation, and with an ED(50) of 2.66 x 10(-13) mol was more potent than any other vasodilator agent yet examined on this vascular bed.5 Bradykinin injected intraportally at doses up to 10 times those which were maximal on the arterial circuit did not alter the calculated HPVR.6 5-HT injected intra-arterially caused weak and variable rises in HAVR, indicating vasoconstriction. The maximum rise in HAVR was much less than that attained with noradrenaline in the same preparations.7 5-HT injected intraportally caused dose-dependent rises in HPVR indicating portal constriction at doses above 15-100 mug: in some experiments small doses of 5-HT resulted in reductions in calculated HPVR.8 Histamine has previously been shown to cause hepatic arterial vasodilatation: by intraportal injection, it caused dose-dependent rises in HPVR.9 In order to examine the receptors responsible for the effects of histamine, dose-response curves were constructed before and after mepyramine and metiamide.10 On the hepatic arterial vascular bed, metiamide did not antagonize the vasodilator effects of intra-arterial histamine, but these effects were antagonized by mepyramine.11 Similarly on the hepatic portal bed, the rises in HPVR due to histamine were antagonized by mepyramine but not by metiamide.12 The effects of histamine on both the hepatic arterial and portal venous vascular beds of the dog are therefore mediated predominantly by histamine H(1)-receptors.     

Abnormalities of Doppler waveform of the hepatic veins in patients with chronic liver disease: correlation with histologic findings

OBJECTIVE: Changes in the Doppler waveform of the hepatic veins are associated with chronic liver disease, particularly cirrhosis. We correlated abnormalities in Doppler waveforms of hepatic veins with histologic findings in the liver to determine the accuracy of Doppler imaging in the detection of cirrhosis. SUBJECTS AND METHODS: Fifty-two patients with chronic hepatitis C were examined prospectively and blindly by two sonographers. In the same session, a liver biopsy specimen was obtained from each patient and submitted to three independent pathologists for conventional interpretation and for grading of severity according to a predetermined scoring system. Duplex sonography of the hepatic veins was also performed in 50 control subjects. RESULTS: Abnormal hepatic vein waveforms were detected in 12 of 16 patients with cirrhosis and in eight of 36 patients without cirrhosis. However, histologic examination of the biopsy specimens showed that only two of the eight patients without cirrhosis had no significant abnormalities, other than mild portal inflammation. Abnormal waveforms were seen in no control subjects. We found a correlation between fibrosis and steatosis and abnormalities in the Doppler waveform of the hepatic veins (r = .50, p < .001). Portal inflammation, intralobular degeneration, and necrosis did not correlate with an abnormal waveform. CONCLUSION: Duplex sonography of the hepatic veins may be useful for studying liver disease associated with fibrosis and steatosis. In patients with well-compensated liver disease, flattening of the Doppler waveform of the hepatic vein suggests the presence of cirrhosis.     

CaBPs and other immunohistochemical markers of the human vomeronasal system: a comparison with other mammals

After more than two centuries of almost sporadic inquiry as to the existence and function of the human vomeronasal system (VNS), the last decade has seen a resurgent interest in it. The principal question vexing many laboratories is whether adult humans retain the VNS that clearly develops during fetal growth. Additional questions are whether the structurally defined fetal VNS has any function role, and if this structure and function extend into postnatal life. One research tool that has been successfully used to identify key components of the mammalian VNS has been immunohistochemistry (IHC). This technique has clearly defined the vomeronasal receptor neurons in the vomeronasal organ, the vomeronasal nerve that projects into the central nervous system, and the target of this nerve, the accessory olfactory bulb. This review will discuss immunohistochemical studies that have identified these features in the mammalian VNS, and relate them to structural and IHC studies of the fetal and adult human VNS. Suggestions as to future studies to clarify the status of the human VNO also are offered.     

Uptake of [3H]dopamine in isolated chromaffin cells of the mouse: modulation by intra- and extra-adrenal peptides and other secretagogues

The effects of intra- and extra-adrenal peptides on [3H]dopamine uptake in adrenal chromaffin cells of the mouse were examined in vitro. Dopamine uptake was inhibited by acetylcholine, high potassium, reserpine, imipramine and desmethylimipramine as was in noradrenaline uptake. Among the intra-adrenal peptides, vasoactive intestinal peptide (VIP, 100 pmol/l) and neurotensin inhibited [3H]dopamine uptake by approximately 25%. Somatostatin, enkephalin, and neuropeptide Y did not cause any significant inhibition. An extra-adrenal peptide, bradykinin, inhibited the uptake while angiotensin II showed no significant effect. Intra-adrenal peptides which cause catecholamine secretion inhibit catecholamine uptake probably to extend its effect. Extra-adrenal peptide which causes catecholamine secretion also inhibits catecholamine uptake.     

Microdissection of peripheral nerve: collagen and lipid distribution with morphological correlation

Peripheral nerve is a complex tissue composed of endoneurial fascicles surrounded by perineurium and epineurium. We separated endoneurium from peri- and epineurium in human sural nerves by "endoneurial plucking", a method of microdissection. Endoneurial contents (axons, myelin sheaths, Schwann cells, vessels, and interstitial collagen) were cleanly separated in high yield from enveloping connective tissue, by both microscopic and biochemical criteria. Most of the nerve sulfatide and unesterified sterol was found in the endoneurial fraction while most of the collagen was in the peri-epineurial fraction. This microdissection method should prove useful in biochemical investigations of peripheral nerve.     

Vector manometric study of the sphincter of Oddi in the dog: functional and morphological correlation

BACKGROUND: Clinical use of cyclosporine (CsA) is limited by its known nephrotoxicity. Parathyroid hormone (PTH)-related protein (PTHrP) increases after acute renal ischemia and stimulates proliferation of renal cells in culture. Herein, we have examined whether the renal expression of PTHrP and its PTH/PTHrP receptor is affected by chronic CsA nephrotoxicity. METHODS: Rats were randomly assigned to receive daily intramuscular injections of either CsA (25 mg/kg) or the same volume of the vehicle olive oil (control) for 3 weeks. At this time interval, under ether anesthesia, rat blood and kidneys were obtained for analytical determinations, and total RNA isolation or immunohistochemistry, respectively. RESULTS: Serum urea was 11+/-2 and 6+/-1 mmol/L (P < 0.01) in CsA-treated and control rats, respectively. We found that PTH/PTHrP receptor mRNA was unchanged, but PTHrP mRNA, and also transforming growth factor-beta1 mRNA expression as positive control, was about twofold increased in the kidney of CsA-treated rats. This was accompanied by increased PTHrP immunostaining in renal cortical tubules, associated with tubule vacuolation. CONCLUSION: This study demonstrates an up-regulation of PTHrP, associated with chronic CsA-induced nephrotoxicity. Our findings support a role for PTHrP in the CsA-injured kidney.     

Studies on the distribution of cholinesterases: activity in the human and dog heart

The distribution and postnatal variation of cholinesterase (ChE) activity were studied in 25 human and 25 dog hearts. The observed distribution pattern is remarkably constant, In dog hearts, the pattern is as follows: sinus node (SN) greater than left atrium (LA) greater than right atrium (RA) greater than right ventricle (RV) congruent to left ventricle (LV). The average acetylcholinesterase (AcChE) activities as expressed in international units per g wet tissue are: 1.66 (SN), 1.14 (LA), 0.70 (RA), 0.22 (RV), and 0.21 (LV). In human hearts, the AcChE distribution follows the pattern of RA greater than LA greater than RV congruent to LV with corresponding average activities of 1.70, 1.38, 0.51, and 0.44 IU. The postnatal variation of ChE activity is most pronounced in the RS of the heart in both species. The average AcChE activity in the RA of the newborn puppies is 0.51 IU as compared with 2.27 IU in newborn infants. In the adult heart, however, the average atrial AcChE activity is nearly identical (1.02 IU) in both species. An additional difference is the large (34-64%) contribution of butyrylcholinesterase (BuChE) to the total activity in dog hearts whereas the contribution of BuChE is small (7-15%) in human hearts.     

Examination of veins with magnetic resonance imaging and it's place in the diagnostic algorithm of patients with deep venous thrombophlebitis]

The aim of our study was to show the results of the Magnetic Resonance Angiography of pelvis and lower extremities. 21 patients with clinical signs of venous thrombosis were examined. The obtained results show the MRA is very useful in evaluation of venous flow.     

Alteration of laminin production in small-cell lung carcinoma: possible correlation with the absence of the basement membrane

The amount of protein recommended to minimise N loss in critically ill patients receiving total parenteral nutrition (TPN) varies in the literature. Therefore, we studied the effect of increased protein intake on the N balance, administering TPN with either 1.2 g protein/kg/day (low N diet) or 1.8 g protein/kg/day (high N diet). Fifteen mechanically ventilated critically ill patients were studied in a surgical intensive care unit. After at least two days of standard TPN, patients were randomly assigned to either the low or the high N diet. Ten patients were studied on the low N diet and 11 on the high N diet; 6 patients were studied on both diets. Nonprotein energy was supplied according to estimated energy requirements. For five consecutive days, the N balance was measured daily. Total urinary nitrogen (TUN) was analysed using the Kjeldahl method. There was no difference in N balance between the groups. On the low N diet, N balance was -0.113 +/- 0.088 and on the high N diet -0.113 +/- 0.109 g N/kg/day. In patients studied twice, N balance was -0.087 +/- 0.054 and -0.050 +/- 0.060 g N/kd/day respectively. Results of a previous pilot study showed that in 20 similar patients the N balance became 80% less negative (from -5.7 +/- 5.1 to -1.1 +/- 8.2 g N/day) when protein intake was increased from 0.9 to 1.5 g/kg/day. Since these results are consistent with other studies, we conclude that the optimal range of protein supply in this type of critically ill patients is approximately 1.1-1.5 g protein/kg/day.     

IgM anti-phenolic glycolipid-I antibody measurements from skin-smear sites: correlation with venous antibody levels and the bacterial index

Measurements of anti-phenolic glycolipid-I antibodies were made in 200 matched samples of capillary blood from the skin-smear site, venous blood collected on filter paper, and sera. A close correlation among the three samples was observed and a weaker correlation among the antibody levels and the average and skin-smear bacterial index. Capillary blood from the skin-smear site had a consistently higher level of antibodies in each sample than did the sera. The collection of capillary blood from skin-smear sites is a convenient and economical method of obtaining samples for serology and for measuring local antibody levels, and it may be more sensitive than measurements of antibodies in sera.     

Structural determinants for binding to CGRP receptors expressed by human SK-N-MC and Col 29 cells: studies with chimeric and other peptides

Structure-activity relationships for the binding of human alpha-calcitonin gene-related peptide 8-37 (halphaCGRP8-37) have been investigated at the CGRP receptors expressed by human SK-N-MC (neuroblastoma) and Col 29 (colonic epithelia) cells by radioligand binding assays and functional assays (halphaCGRP stimulation of adenylate cyclase). On SK-N-MC cells the potency order was halphaCGRP8-37 > halphaCGRP19-37 = AC187 > rat amylin8-37 > halpha[Tyr0]-CGRP28-37 (apparent pKBs of 7.49+/-0.25, 5.89+/-0.20, 6.18+/-0.19, 5.85+/-0.19 and 5.25+/-0.07). The SK-N-MC receptor appeared CGRP1-like. On Col 29 cells, only halphaCGRP8-37 of the above compounds was able to antagonize the actions of halphaCGRP (apparent pKB=6.48+/-0.28). Its receptor appeared CGRP2-like. halpha[Ala11,18]-CGRP8-37, where the amphipathic nature of the N-terminal alpha-helix has been reduced, bound to SK-N-MC cells a 100 fold less strongly than halphaCGRP8-37. On SK-N-MC cells, halphaCGRP8-18,28-37 (M433) and mastoparan-halphaCGRP28-37 (M432) had apparent pKBs of 6.64+/-0.16 and 6.42+/-0.26, suggesting that residues 19-27 play a minor role in binding. The physico-chemical properties of residues 8-18 may be more important than any specific side-chain interactions. M433 was almost as potent as halphaCGRP8-37 on Col 29 cells (apparent pKB=6.17+/-0.20). Other antagonists were inactive.