重型乙型肝炎患者原位肝移植围手术期全身氧代谢的变化

目的 探讨重型乙型肝炎（乙肝）与其他肝病患者原位肝移植围术期全身氧代谢变化的特点。方法 12例重型乙肝患者为试验组。10例其他肝病患者为对照组。以咪唑安定、异丙酚、芬太尼、维库溴铵诱导全麻，术中吸入异氟醚维持麻醉。维库溴铵维持肌松，行改良背驼式原位肝移植术。左桡动脉穿刺测有创动脉压，右颈内静脉穿刺置入漂浮导管。分别于术前、无肝前10min、无肝期25min、新肝期30min和术毕监测动脉和混合静脉血氧分压（PaO2和Pv^-O2）、动脉和混合静脉血氧含量（CaO2和Pv^-O2）及动-静脉血氧含量差（CA-vO2）、氧供（DO2）、氧供指数（DO2I）、氧消耗（VO2）、氧耗指数（VO2I）、氧摄取指数（O2EI）和氧摄取率（O2ER）。结果 ①试验组：与术前相比，无肝前期Pv^-O2上升，Ca-vO2、O2EI、O2ER下降，DO2和VO2无明显变化；无肝期DO2、DO2I、VO2和VO2I均明显下降，DO2、VO2分别下降43％和21％，O2EI和O2ER均明显上升；新肝期PvO2上升，DO2和DO2I明显上升。VO2和VO2I回升至术前水平；术毕时DO2和DO2I依然高于术前水平。②对照组：无肝前期PvO2上升。DO2和VO2无明显变化，O2EI和O2ER下降；无肝期DO2、DO2I、VO2和VO2I均明显下降，DO2下降25％，VO2则下降12％；新肝期PvO2上升，Ca-vO2下降，DO2、DO2I明显上升，VO2和VO2I回升至术前水平；术毕时DO2和DO2I依然高于术前水平。结论肝移植围术期中，全身DO2变化大于VO2变化；重型乙肝患者的全身DO2和VO2变化较其他肝病患者剧烈。     

Role of endothelium in adenosine receptor-mediated vasorelaxation in hypertensive rats

The present study was designed to investigate the role of endothelium derived relaxing factor nitric oxide (NO) in adenosine A2 receptor mediated vasorelaxation in normotensive (WKY) and hypertensive (SHR) rat aortic ring preparations. Adenosine analogues, 2-chloroadenosine (CAD) and 5-ethylcarboxamidoadenosine (NECA) produced concentration-dependent (10(-9)-10(-4) M) relaxation in phenylephrine (1 x 10(-6) M) precontracted vascular rings, which was significantly shifted to the right in SHR compared to WKY rats. Endothelium removal attenuated CAD and NECA relaxation responses in both SHR and WKY and abolished the difference in relaxation between SHR and WKY vascular tissues. The relaxation response to CAD was antagonised by adenosine A2 receptor antagonist, 8-sulfophenyltheophylline (8-SPT, 50 x 10(-6) M). The antagonism by 8-SPT was lower in SHR as compared to WKY tissues. L-monomethylarginine (L-LMMA) (30 x 10(-6) M) significantly shifted the CAD relaxation to the right, which was reversed by the addition of L-arginine (100 x 10(-6) M) in both SHR and WKY rats. However, the rightward shift by L-NMMA was smaller in SHR compared to WKY vascular tissues. Vasorelaxation response to acetylcholine (1 x 10(-6) M) was significantly inhibited (50%) in SHR rings compared to WKY. The relaxation produced by sodium nitroprusside (10(-9)-10(-5) M) in endothelium-intact and -denuded aortic rings showed no difference between SHR and WKY. Isoproterenol produced concentration-dependent (10-9-10-5 M) relaxation, which was shifted to the right in SHR compared to WKY rings with an intact endothelium, while the removal of endothelium abolished the difference in the response between SHR and WKY. The results suggest: (i) adenosine A2 receptors mediate vasorelaxation in part through endothelium possibly by releasing nitric oxide (NO); (ii) the impairment of endothelium may be one of the factors for the attenuation of adenosine receptor and receptor-mediated responses in SHR.     

Signaling in H2O2-induced increase in cell proliferation in Barrett's esophageal adenocarcinoma cells

Mechanisms whereby acid reflux may accelerate the progression from Barrett's esophagus (BE) to esophageal adenocarcinoma (EA) are not fully understood. We have previously shown that NADPH oxidase NOX5-S generates reactive oxygen species (ROS) when Barrett's metaplastic cells are exposed to acid. Besides metaplastic cells, other H(2)O(2)-producing cells (e.g., inflammatory cells) present in BE mucosa may produce additional ROS, which may also affect metaplastic cells contributing to esophageal tumorigenesis. In this study, we investigate whether exogenous H(2)O(2) stimulates cell proliferation by increasing NOX5-S expression. Low dose (10(-13) M) of H(2)O(2) significantly increased thymidine incorporation, NOX5-S mRNA, and protein expression in a Barrett's EA cell line FLO. H(2)O(2)-induced increase in NOX5-S expression was significantly inhibited by knockdown of nuclear factor (NF)-κB1 p50 with p50 small interfering RNA (siRNA) in EA cell lines FLO and OE33. H(2)O(2) significantly increased p65 phosphorylation and the luciferase activity in FLO cells transfected with a NF-κB activation reporter plasmid pNF-κB-Luc. H(2)O(2)-induced increase in luciferase activity in FLO cells was significantly decreased by knockdown of extracellular signal-regulated kinase 2 (ERK2) mitogen-activated protein kinase (MAPK). Overexpression of p50 and p65 remarkably increased the luciferase activity in FLO cells transfected with a NOX5-S reporter plasmid NOX5-LP. In addition, H(2)O(2)-induced thymidine incorporation in FLO cells was significantly decreased by the MAPK kinase 1/2 inhibitor 2'-amino-3'methoxyflavone (PD98059) and ERK2 siRNA but not by ERK1 siRNA. Likewise, H(2)O(2)-induced increase in NOX5-S expression was significantly decreased by ERK2 siRNA in FLO and OE33 cells. We conclude that a low dose of H(2)O(2) increases cell proliferation. H(2)O(2)-induced increase in cell proliferation may depend on sequential activation of ERK2 MAPK, NF-κB1 p50, and NOX5-S.     

Evolution of erythromycin resistance in Streptococcus pneumoniae in Italy

OBJECTIVES: To evaluate erythromycin resistance in recent invasive isolates of Streptococcus pneumoniae in Italy, to study the phenotypic and genotypic characteristics of the isolates, and to compare data with those obtained in a previous survey. METHODS: Invasive pneumococcal isolates were obtained from 56 laboratories throughout the country, in 2001-2003. Isolates were serotyped and antimicrobial susceptibilities determined by Sensititre panels and Etest. A new PCR was performed to detect erythromycin resistance genes. Typing methods for selected erythromycin-resistant isolates included PFGE and multilocus sequence typing (MLST). RESULTS: One hundred and fifty-five isolates out of 444 (34.9%) were resistant to erythromycin: 95 isolates (21.4%) carried erm(B), 56 (12.6%) carried mef(A) and three carried both genes. One isolate, carrying neither erm(B) nor mef(A), showed a point mutation in domain V of the 23S rRNA genes. The mef(A)-positive isolates carried subtype mef(A) (47 isolates), subtype mef(E) (nine isolates), and both subtype mef(E) and erm(B) (three isolates). All subtype mef(A) strains, except two, belonged to serotype 14, appeared to be clonally related by PFGE and related to the England14-9 clone by MLST. The two isolates belonging to other serotypes showed different genetic backgrounds. CONCLUSIONS: Erythromycin resistance in S. pneumoniae has increased in the last few years in Italy. erm(B) is still the predominant resistance determinant; however, the increase in erythromycin resistance (34.9% versus 28.8% of the previous years) is mainly due to an increase in the proportion of isolates carrying the efflux pump mef(A), whereas the proportion of isolates carrying erm(B) has not changed.     

Sexual dimorphism in phencyclidine in vitro metabolism and pharmacokinetics in rats.

Studies were conducted to determine the differences in phencyclidine (PCP) in vitro metabolism and pharmacokinetics in female and male Sprague-Dawley (SD) rats. Formation rates of five major PCP metabolites in liver microsomes were significantly higher (p <.05) in males compared with females in three different rat strains (SD, Fischer 344, and Dark Agouti). In addition, the formation rate for an irreversibly bound PCP metabolite in males was the second highest of the six metabolites measured in these studies. However, the liver microsomes from the females produced essentially no metabolite binding in any strain. To determine the in vivo consequences of these in vitro metabolism results, we determined PCP's pharmacokinetic profile in female SD rats after a pharmacologically active i.v. dose of PCP (1 mg/kg) and then compared these data with the pharmacokinetic profile in male SD rats. The value for PCP systemic (and nonrenal) clearance was more than 45% lower (p <.05) in female rats. In addition, the terminal elimination T(1/2) was significantly longer (p <.05) in the female rats (5.5 versus 3.4 h, respectively). Because the initial serum concentration, volume of distribution at steady state, and renal clearance were not significantly different between the sexes, the longer half-life was attributed directly to a decreased ability of females to metabolize the drug. Consequently, these pharmacokinetic data suggest pharmacological differences in PCP effects between female and male rats are due primarily to a decreased ability of female rats to metabolize the drug.     

Exploring variation in pressure ulcer prevalence in Sweden and the USA: benchmarking in action

Aim To compare overall unit‐level pressure ulcer (PU) prevalence, hospital‐acquired pressure ulcer (HAPU) prevalence and prevention strategies, as well as nurse staffing and workload in two hospitals in Sweden with data from the USA. Methods Medical and surgical units in a university hospital and a general hospital in Sweden were compared with 207 hospitals in the USA participating in the Collaborative Alliance for Nursing Outcomes (CALNOC) benchmarking registry. All adult inpatients in university hospital (n = 630), general hospital (n = 253) and CALNOC hospitals (n = 3506) were included in the study. Outcome indicators were pressure ulcer prevalence for all types (PU) and HAPU prevalence, specifically. Process indicators were risk assessment and PU prevention strategies. Structure indicators were nurse staffing (hours of care, and skill mix) and workload (admissions, discharges and transfers). Results The prevalence of PU (categories 1–4) was 17.6% (university hospital) and 9.5% (general hospital) compared with 6.3–6.7% in the CALNOC sample. The prevalence of full thickness HAPU (categories 3 and 4) was 2.7% (university hospital) and 2.0% (general hospital) compared with 0–0.5% in the CALNOC sample. Risk and skin assessment varied between 6% and 60% in the Swedish hospitals compared with 100% in the CALNOC sample. Total hours per patient day were 8.4 in both Swedish hospitals and 9.5 to 9.8 in the CALNOC hospitals Conclusions The findings suggest a link between processes of care and outcomes that is exciting to observe internationally and suggest the opportunity to expedite performance improvement through global benchmarking. Using HAPU as a complement to point prevalence of PU in Sweden has revealed this indicator as a more valid measure for patient care quality.     

Phosphodiesterase isozymes involved in regulation of formula secretion in isolated mouse stomach in vitro

(+/-)-(E)-4-Ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide] (NOR-3), a nitric-oxide (NO) donor, is known to increase HCO(3)(-) secretion in rat stomachs, intracellularly mediated by cGMP; yet, there is no information about the phosphodiesterase (PDE) isozyme involved in this process. We examined the effects of various isozyme-selective PDE inhibitors on the secretion of HCO(3)(-) in the mouse stomach in vitro and the type(s) of PDE isozymes involved in the response to NO. The gastric mucosa of DDY mice was stripped of the muscle layer and mounted on an Ussing chamber. HCO(3)(-) secretion was measured at pH 7.0 using a pH-stat method and by adding 2 mM HCl. NOR-3, 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP), and various PDE inhibitors were added to the serosal side. Vinpocetine (PDE1 inhibitor) or zaprinast (PDE5 inhibitor) was also added serosally 30 min before NOR-3 or 8-Br-cGMP. Both NOR-3 and 8-Br-cGMP stimulated HCO(3)(-) secretion in a dose-dependent manner, and the response to NOR-3 was significantly inhibited by methylene blue. Likewise, the secretion induced by NOR-3 or 8-Br-cGMP was significantly attenuated by 6-((2S,3S)-3-(4-chloro-2-methylphenylsulfonylaminomethyl)-bicyclo(2.2.2)octan-2-yl)-5Z-hexenoic acid (ONO-8711), the PGE receptor (EP)1 antagonist, as well as indomethacin and potentiated by both vinpocetine and zaprinast at doses that had no effect by themselves on the basal secretion, whereas other subtype-selective PDE inhibitors had no effect. NOR-3 increased the mucosal PGE(2) content in a methylene blue-inhibitable manner. These results suggest that NO stimulates gastric HCO(3)(-) secretion mediated intracellularly by cGMP and modified by both PDE1 and PDE5, and this response is finally mediated by endogenous PGE(2) via the activation of EP1 receptors.     

Bone remodeling markers in the detection of bone metastases in prostate cancer

BACKGROUND: Early detection of bone metastases in prostatic carcinoma is very useful in treatment and prognosis of the disease. The aim of this work was to evaluate the sensitivity and specificity of a group of bone markers in order to discriminate between prostate carcinoma patients without (M(0)) and with (M(1)) bone metastases. METHODS: Sixty-seven non-treated patients with: benign prostate hyperplasia (n=21), prostatic carcinoma in several stages without bone metastases (T(X)M(0)) (n=31) and with bone metastases (T(X)M(1)) (n=15) were studied. The following markers were studied: (A) bone formation: (1) serum bone alkaline phosphatase, IRMA (Tandem Ostase, Beckman); (2) serum procollagen I amino-terminal propeptide (PINP), RIA (Orion Diagnostica); (B) bone resorption: (1) urinary collagen I amino-terminal telopeptide (NTX), ELISA (Ostex); (2) collagen I carboxy terminal telopeptide (CTX): (2A) urinary alpha-CTX, RIA (Osteometer), (2B) serum beta-CTX, Elecsys (Roche); (3) collagen I cross-linked carboxy terminal telopeptide (ICTP), RIA (Orion Diagnostica). RESULTS: Levels of all bone markers were significantly higher in group M(1) than in group M(0). A complete separation of groups M(0) and M(1) was achieved with PINP and beta-CTX (100% sensitivity and specificity). CONCLUSIONS: These results support the use of PINP or beta-CTX as a tool to confirm the presence or absence of bone metastases in the first staging of prostatic carcinoma patients.     

Improvement in glycemic control over 11 years in patients monitored for diabetes in one county.

BACKGROUND: Hemoglobin A(1c) (HbA(1c)) has been used in controlled trials for the last 10 years but has never been evaluated in clinical practice as an effective parameter for clinical outcome. We investigated the trend for glycemic control over 11 years in one county of 350,000 citizens. METHODS: We studied 226,382 HbA(1c-DCCT-aligned) from 39,455 patients in whom routine monitoring for diabetes was initiated in 1993, 1996, or 2001. The trend in glycemic control was investigated in groups by probit plots, and in individual patients by target plots. RESULTS: From 1993 to 2001, the number of HbA(1c) measurements increased three-fold. The number of new monitoring series increased from 0.22% to 0.27% of the county population, and the number of patients monitored using HbA(1c) as a proxy for diabetes increased from 0.5% to 1.5%. A proportional reduction in high HbA(1c) concentrations of 5% was identified in the 1993 group, compared to 15% in the 1996 group, and 20% in the 2001 group. The percentage of patients with diabetic first HbA(1c) experiencing normalization increased from 8% to 30% for males and from 9% to 24% for females (1993-2001). The percentage of HbA(1c) concentrations that were not normalized decreased from 78% to 53% for males and from 83% to 59% for females (1993-2001). The median HbA(1c) at initiation of monitoring decreased from 8.7% in 1993 to 7.5% in 2001 (p < 10(-5)). The number of normal first HbA(1c) results in monitoring series increased from 7% to 17% for males and from 8% to 22% for females. Up to 10% of subjects developed diabetic concentrations during monitoring. CONCLUSION: On average, patients with diabetic first HbA(1c) concentrations (> or =6.62%) showed an improvement in glycemic control from 5% in 1993 to 20% in 2001. High concentrations were easiest to reduce. In patients with originally diabetic HbA(1c) levels, 66% on average showed improved glycemic control in the 2001 series compared to 50% in the 1993 series. An average of 6% (1993) vs. 9% (2001) with originally normal HbA(1c) levels showed an upward trend inHbA(1c) levels. Median HbA(1c) at initiation of monitoring decreased from 8.7% in 1993 to 7.5% in 2001 (p < 10(-5)). The incidence of new cases was constant.     

Mechanisms of Hydrogen Peroxide-Induced Increase in Intracellular Calcium in Cardiomyocytes

BACKGROUND: Hydrogen peroxide (H(2)O(2)) in high concentrations has been implicated in heart dysfunction attributable to ischemia-reperfusion. Although H(2)O(2) is also known to increase the intracellular concentration of Ca(2+) ([Ca(2+)](i)) in cardiomyocytes, the mechanisms for such a change are not clear. In this study, the sources and mechanisms of increase in [Ca(2+)](i) caused by high concentrations of H(2)O(2) in cardiomyocytes were explored. METHODS AND RESULTS: Cardiomyocytes were isolated from adult male Sprague-Dawley rats. Cell viability was examined by trypan blue exclusion test. [Ca(2+)](i) was measured by employing cell suspension at room temperature and Fura-2 fluorescence technique. Incubation of cells with 0.25-l mmol/L H(2)O(2) increased [Ca(2+)](i) in a time- and concentration-dependent manner. Catalase attenuated the H(2)O(2)-induced increase in [Ca(2+)](i) significantly, whereas mannitol showed no effect. Neither the presence of verapamil, a sarcolemmal Ca(2+) channel blocker, nor the removal of Ca(2+) from the medium produced any significant reduction in the H(2)O(2)-induced increase in [Ca(2+)](i). Conversely, treatment of cardiomyoctes with staurosporin, a protein kinase C inhibitor, thapsigargin, a sarcoplasmic reticulum Ca(2+)-pump adenosine triphosphatase inhibitor, as well as ryanodine, a sarcoplasmic reticulum Ca(2+)-release channel blocker, markedly prevented the 0.5-mmol/L H(2)O(2)-induced increase in [Ca(2+)](i). The responses of cardiomyoctes to H(2)O(2) and other Ca(2+)-mobilizing agents, such as KCl or adenosine triphosphate, were additive. No changes in cardiomyocyte viability were seen on incubation with 0.5 and 1 mmol/L H(2)O(2). Perfusion of the isolated heart with H(2)O(2) (0.1-0.5 mmol/L) depressed the left ventricular developed pressure, rate of contraction, and rate of relaxation, whereas the left ventricular end-diastolic pressure was increased. CONCLUSIONS: These results indicate that formation of H(2)O(2) under pathophysiological conditions such as ischemic heart disease may induce changes in Ca(2+) homeostasis in cardiomyocytes and may induce contractile dysfunction. Furthermore, the sarcoplasmic reticulum involving a protein kinase C-mediated mechanism appears to be the main site of action of H(2)O(2) in cardiomyocytes.     

Causes of death in patients with rheumatic diseases in Moscow

AIM: To study causes of death of rheumatic patients in Moscow. MATERIAL AND METHODS: Autopsy protocols for 1999-2002 were analysed for two pathoanatomic departments of Moscow. RESULTS: Rheumatic diseases were detected in 165 cases (2.0% of overall number of autopsies in these departments). Rheumatic heart disease (RHD) was stated in 99 (60.0%) cases, rheumatic fever relapse (RFR) in 4 (2.4%), rheumatoid arthritis (RA) in 28 (17.0%), systemic lupus erythematosus (SLE) in 8 (4.8%), systemic sclerosis (SS) in 3 (1.8%), ankylosing spondilarthritis (AS) in 2 (1.2%), systemic vasculitis (SV) in 2 (1.2%), osteoarthrosis in 11 (7.3%), gout in 3 (1.8%), polymyositis in 1 (0.6%). RHD patients died of decompensated circulation (DC) (54%), acute heart failure (AHF) (14%), thromboembolism (TE) (6%), other causes (26%). RF patients died of TE (n = 2), DC (n = 1), AHF (n = 1). Out of 28 RA patients, 5 patients died of secondary amyloidosis, 3 of DC, 7 of AHF, 1 of TE, 5 of infectious complications, 7 of other causes. SLE patients died of uremia (n = 2), acute adrenal failure (n = 1), infectious complications (n = 2), AHF (n = 2), brain edema (n = 1). CONCLUSION: Among rheumatic diseases, rheumatic heart valve disease was most severe as it caused the highest mortality. Cardiovascular pathology caused death in most of the rheumatic patients.     

Myocardial injury in meningococcus-induced purpura fulminans in children

OBJECTIVES: To assess the incidence of myocardial ischemia in meningococcus-induced purpura fulminans in pediatric patients, to compare troponin I (cTnI) levels with changes in electrocardiogram (ECG) and to evaluate whether cTnI is related to myocardial function and contractility, to severe acquired anticoagulant deficiency and to the severity of disease. METHODS: Twenty-two patients with acute meningococcemia, supported with inotropes or vasoactive agents, were studied, Blood samples for the determination of serum cTnI and conventional myocardial ischemia and coagulopathy markers were drawn daily. Measurements of cardiac index (CI), ejection (EF) and shortening fractions (SF) and ECGs were performed daily. RESULTS: The Leclerc score, the Neisseria sepsis index (NESI) and the pediatric risk of mortality (PRISM) score predicted a mean mortality rate of 34%, 27% and 23%, respectively. Four patients died (18%). Five patients (23 %) presented with myocardial ischemia. Their ECG ischemic changes were associated with pathologically high cTnI levels (1.93 +/- 0.13 vs 0.18 +/- 0.08 ng/ml, p < 0.001 for patients with or without ischemic changes) and depressed myocardial contractility (mean difference +/- SE -14 +/- 5%, p = 0.01, for the EF and -7.4 +/- 3, p < 0.02, for the SF). High cTnI values were significantly correlated to low protein C (PC) (p < 0.0001), factor VIII (p < 0.04) and antithrombin III (AIII, p = 0.01) levels, but not to the PRISM, Leclerc or the NESI scores. Means of AIII, VII, and especially of VIII, and PC, were significantly lower in ischemic than in non-ischemic patients, although severity scoring systems and inotropic support did not differ between the two groups. Survivors tended to significantly higher PC (p < 0.01) and factor VIII levels (p = 0.001) than non-survivors and, also, to lower levels of cTnI (p = 0.05) and CPK-MB (p < 0.05), while in meningococcal shock. CONCLUSIONS: The incidence of myocardial ischemia is increased in acute meningococcemia in pediatric patients and correlates with myocardial dysfunction. High cTnI is associated with severe coagulopathy, but not with clinical prognostic scores or inotropic support. Early recognition of myocardial injury, myocardial support and early replacement therapy with PC, AIII, factor VIII or fibrinogen might improve outcome in acute meningococcemia in children.     

Indices of proteolysis in evaluation of resorption in bone tumors

Activities of acid and alkaline phosphatases, collagenase, cathepsin D, trypsin-like proteinases, alpha(1)-proteinase inhibitor (alpha(1)-PI), alpha(2)-macroglobulin (alpha(2)-MG) were measured in blood plasma and tumor tissue of patients with giant-cell tumor of the bone (GCTB) and bone chondrosarcoma. These tumors differed by enzymatic activities. GCTB is characterized by increased activity of alkaline phosphatase, while in chondrosarcoma tissue the activities of collagenase and cathepsin D were the highest. Activities of acid phosphatase, collagenase, trypsin-like proteinases were increased in the plasma of patients with both tumors; alpha(1)-PI/alpha(2)-MG ratio was increased. Bone resorption parameters correlated with proteolysis inhibitors. Activities of collagenase and acid phosphatase were increased in tumor tissue and plasma in the presence of low activities of alpha(2)-MG and increased alpha(1)-PI/alpha(2)-MG index, which seems to require special attention during the postoperative period.     

Chymase-dependent conversion of Big endothelin-1 in the mouse in vivo

The aim of this study was to identify the role of chymase in the conversion of exogenously administered Big endothelin-1 in the mouse in vivo. Real-time polymerase chain reaction analysis detected mRNA of mucosal mast cell chymases 4 and 5, endothelin-converting enzyme 1a, and neutral endopeptidase 24.11 in pulmonary, cardiac, and aorta homogenates derived from C57BL/6J mice, with the latter tissue expressing the highest levels of both chymase isoforms. Furthermore, hydrolysis of a fluorogenic peptide substrate, Suc-Leu-Leu-Val-Tyr-7-amino-4-methylcoumarin, was sensitive to the chymase inhibitors Suc-Val-Pro-Phe(P)(OPh)(2) (200 microM) and chymostatin [(S)-1-carboxy-2-phenylethyl]-carbamoyl-alpha-[2-iminohexahydro-4(S)-pyrimidyl]-(S)-Gly-X-Phe-al, where X can be the amino acid Leu, Val, or Ile) (100 microM) in supernatants extracted from the same tissue homogenates. In anesthetized mice, Big endothelin-1, endothelin-1 (1-31), and endothelin-1 triggered pressor responses (ED(50)s, 0.67, 0.89, and 0.16 nmol/kg) that were all reduced or potentiated by selective endothelin ET(A) or ET(B) receptor antagonists, respectively, BQ-123 (cyclo[D-Asp-Pro-D-Val-Leu-D-Trp]) or BQ-788 (N-[N-[N-[(2,6-dimethyl-1-piperidinyl)carbonyl]-4-methyl-l-leucyl]-1-(methoxycarbonyl)-D-tryptophyl]-d-norleucine sodium salt), each at 1 mg/kg. The pressor responses to big endothelin-1 were significantly reduced by the neutral endopeptidase inhibitor thiorphan (dl-3-mercapto-2-benzylpropanoylglycine) (1 mg/kg) or the endothelin-converting enzyme inhibitor CGS 35066 [alpha-[(S)-(phosphonomethyl)amino]-3-dibenzofuranopropanoic acid] (0.1 mg/kg). In contrast, the responses to endothelin-1 (1-31) were abolished by thiorphan but unaffected by CGS 35066. In addition, Suc-Val-Pro-Phe(P)(OPh)(2) (20-40 mg/kg) reduced, by more than 60%, the hemodynamic response to big endothelin-1 but not to endothelin-1 (1-31) and endothelin-1. Finally, intravenous administration of big endothelin-1 induced Suc-Val-Pro-Phe(P)-(OPh)(2)-sensitive increases in plasma-immunoreactive levels of endothelin-1 (1-31) and endothelin-1. The present study suggests that chymase plays a pivotal role in the conversion and cardiovascular properties of big endothelin-1 in vivo.     

Cyclooxygenase-2-mediated angiogenesis in carrageenin-induced granulation tissue in rats

The possible participation of cyclooxygenase (COX)-2 in angiogenesis in granulation tissue was analyzed using an air pouch-type carrageenin-induced inflammation model in rats. Injection of carrageenin solution into an air pouch induced gradual increases in the pouch fluid volume and granulation tissue weight as well as angiogenesis in granulation tissue. NS-398 (10-100 microg) inhibited all of these parameters in a dose-dependent manner. NS-398 (100 microg), indomethacin (100 microg), and dexamethasone (10 microg) markedly reduced prostaglandin (PG) E(2) levels in the pouch fluid at day 6. NS-398 and indomethacin did not affect protein levels of COX-1 and COX-2 but dexamethasone significantly reduced the level of COX-2 in granulation tissue at day 6. Protein levels of vascular endothelial growth factor (VEGF) in granulation tissue and in the pouch fluid were higher at day 6 than at day 3, and the levels were decreased by treatment with NS-398 (10-100 microg) in a dose-dependent manner. The inhibitory effects of NS-398 (100 microg) were almost the same as those of indomethacin (100 microg). Dexamethasone (10 microg) also reduced VEGF protein levels in granulation tissue at day 6. To clarify the role of PGE(2) in VEGF production, minced granulation tissue obtained 3 days after carrageenin injection from the indomethacin-treated rats was incubated in the presence of various concentrations of PGE(2). It was shown that VEGF mRNA and protein levels in the minced granulation tissue were increased by PGE(2) in a concentration-dependent manner. These findings suggest that COX-2-derived PGE(2) plays a significant role in angiogenesis in the carrageenin-induced granulation tissue through VEGF formation.     

Role of gamma-aminobutyric acid (GABA)A and GABAB receptors in paraventricular nucleus in control of sympathetic vasomotor tone in hypertension

The paraventricular nucleus (PVN) of the hypothalamus is involved in tonic regulation of sympathetic outflow. Impaired GABAergic control of PVN neurons may contribute to the elevated sympathetic drive in hypertension. In this study, we examined the function of GABA(A) and GABA(B) receptors in the PVN in control of sympathetic nerve activity and arterial blood pressure (ABP) in normotensive and hypertensive rats. Lumbar sympathetic activity (LSNA) and ABP were recorded from anesthetized spontaneously hypertensive rats (SHRs), Sprague-Dawley (SD) rats, and Wistar-Kyoto (WKY) rats. Bilateral microinjection of bicuculline (0.01-0.15 nmol), a GABA(A) receptor antagonist, into the PVN increased LSNA and ABP in normotensive WKY and SD rats in a dose-dependent manner. This response was significantly attenuated in SHRs. Furthermore, the decrease in LSNA and ABP induced by a GABA(A) receptor agonist, muscimol (0.05-1.5 nmol), in the PVN was significantly less in SHRs than in normotensive controls. In contrast, microinjection of the GABA(B) receptor agonist baclofen (0.3-4.5 nmol) into the PVN decreased LSNA and ABP in SHRs. However, in WKY and SD rats, baclofen only decreased LSNA and ABP at the highest dose tested. In addition, blockade of GABA(B) receptors in the PVN with CGP52432 (3-[[(3,4-dichlorophenyl)methyl]amino]propyl]diethoxymethyl)phosphinic acid) (0.15-3.0 nmol) dose-dependently increased LSNA and ABP in SHRs but not in normotensive controls. Collectively, this study provides new evidence that GABA(A) receptor function is attenuated, whereas the function of GABA(B) receptors is enhanced, in the PVN of SHRs.     

Comparison of AIDS in women in rural and urban Georgia.

Through 1990, 308 cases of AIDS had been reported in female residents of Georgia (aged 13 years and older); 77 (25%) were white and 228 (75%) black. The mean age of the white women (43.8 years) was greater than that of the black women (34.5 years). One hundred sixty-six women were from Atlanta (metro Atl), the major metropolitan center in Georgia, and 142 (46%) from other regions of the state (other areas). Blacks represented 74% and 76% of all cases in women in metro Atl and other areas respectively. Of the 308 cases, 178 (58%) were related to intravenous (IV) drug use (99 metro Atl, 79 other areas), including 104 (58%) women who were IV drug users and 74 who were sexual partners of IV drug users. These proportions were similar in the two regions. Among cases related to IV drug use, 85 (86%) women in metro Atl and 69 (87%) women elsewhere in the state were black. The cumulative rate of AIDS in women in metro Atl (14.4/100,000) was twice that of women in the rest of the state (6.7/100,000). The rate for AIDS cases related to IV drug use in black women (27.8/100,000 population) was 19 times that in white women (1.5/100,000). The median survival of all women was significantly greater in metro Atl than in other areas (400 and 296 days respectively), with a difference also in those reported only with Pneumocystis carinii pneumonia (466 and 373 days respectively).     

Metalloproteinase-9 in circulating monocytes in pulmonary hypertension

The role of matrix metalloproteinases (MMPs) in pulmonary hypertension (PH) is complex as MMPs are involved in both the vascular and cardiac remodelling associated with PH. To gain insight into this problem, monocytes were isolated from pulmonary arterial blood in patients suffering from PH, related to chronic obstructive pulmonary disease (n = 6), chronic pulmonary thromboembolism (n = 3) or pulmonary arterial hypertension (n = 8). The severity of PH was associated with decreases in cardiac index (CI) and mixed venous blood oxygen saturation (SO(2)), and an increase in right atrial pressure (). Monocyte pro-MMP-9 content (zymography) was positively correlated with SO(2) (r = 0.73, P < 0.05) and CI (r = 0.66, P < 0.05), and negatively with (r = 0.54, P < 0.05); there was no significant correlation with pulmonary vascular resistance. In conclusion, the pro-MMP-9 content of circulating monocytes was lower in the more severe forms of PH which showed heart failure suggesting that such MMP enzymatic activity reflects heart failure following pulmonary vascular and myocardial remodelling in PH.     

Five polymorphisms in gene candidates for cardiovascular disease in Afro-Brazilian individuals

Associations of polymorphisms in the angiotensin I-converting enzyme (ACE), apolipoprotein B (APOB) and apolipoprotein E (APOE) genes with hypertension and variations in lipid serum levels were evaluated in 184 Afro-Brazilians with a familial history of coronary artery disease (CAD). ACE (Ins/Del) and APOB (Ins/Del, XbaI, and EcoRI) and APOE (HhaI) polymorphisms were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses on agarose, and polyacrylamide gel electrophoresis. Serum lipids were measured by means of routine enzymatic assays. The results showed a high frequency of hypertension (44%) in Afro-Brazilians that was increased in subjects >40 years old and those with a blood mass index (BMI) higher than 25 kg/m(2) (P<0.001). The ACE Del allele was associated with hypertension in men >40 years old (P<0.05). APOE (HhaI) and APOB (XbaI and Ins/Del) polymorphisms were not associated with hypertension or variations in serum concentrations of lipids, while subjects with the APOB E- allele had higher low-density lipoprotein cholesterol (LDL-C) levels than E+ carriers (P<0.05). These results suggest that ACE Ins/Del polymorphism is associated with hypertension, and APOB EcoRI polymorphism is associated with LDL-C variation in Afro-Brazilians.     

Supine fall in lung volumes in the assessment of diaphragmatic weakness in neuromuscular disorders

OBJECTIVE: To determine whether diaphragmatic function can be determined by noninvasive respiratory indices in neuromuscular disease. DESIGN: Vital capacity (VC) and mouth pressure generated during a maximal static inspiratory effort (Pi max) were measured with patients in both sitting and supine positions. SETTING: Rehabilitation hospital. PATIENTS: Twenty-four patients with generalized neuromuscular disease. MAIN OUTCOME MEASURES: Changes in indices from sitting to supine position were compared with invasive diaphragmatic function indices consisting of transdiaphragmatic pressures during maximal sniff (Pdi sniff) and the ratio of gastric pressure (Pga) increases over transdiaphragmatic pressure (DeltaPga/DeltaPdi) during quiet breathing. RESULTS: The fall in VC in the supine position was greater in the 15 patients who had spontaneous paradoxical diaphragmatic motion (DeltaPga/DeltaPdi < 0) than in the 9 patients who did not. Specificity and sensitivity of a greater than 25% supine fall in VC for the diagnosis of diaphragmatic weakness (DeltaPga/DeltaPdi < 0 and/or Pdi sniff < 30cmH2O) were 90% and 79%, respectively. Stepwise multiple regression analysis of Pdi sniff showed that both the supine fall in VC and Pi max were associated with diaphragmatic weakness (R(2) =.66; p <.0001). These factors contributed 52% and 14% of the Pdi sniff variance, respectively. CONCLUSIONS: Simple VC measurement in the sitting and supine positions may be helpful in detecting severe or predominant diaphragmatic weakness.