OBJECTIVE

The characterization of primary HIV infection by the analysis of serial plasma samples from newly infected persons using multiple standard viral assays.

METHODS

A retrospective study involving two sets of archived samples from HIV-infected plasma donors. (A) 435 samples from 5<em>1</em> donors detected by anti-HIV enzyme immunoassays donated during <em>1</em>984-<em>1</em>994; (B) <em>1</em>45 specimens from 44 donors detected by p24 antigen screening donated during <em>1</em>996-<em>1</em>998.

METHODS

Two US plasma products companies.

METHODS

The timepoints of appearance of HIV-<em>1</em> markers and viral load concentrations during primary HIV infection.

RESULTS

The pattern of sequential emergence of viral markers in the 'A' panels was highly consistent, allowing the definition and estimation of the duration of six sequential stages. From the 'B' panels, the viral load at p24 antigen seroconversion was estimated by regression analysis at <em>1</em>0 000 copies/ml (95% CI 2000-93 000) and the HIV replication rate at 0.35 log copies/ml/day, corresponding to a doubling time in the preseroconversion phase of 20.5 h (95% CI <em>1</em>8.2-23.4 h). Consequently, an RNA test with 50 copies/ml sensitivity would detect HIV infection approximately 7 days before a p24 antigen test, and <em>1</em>2 days before a sensitive anti-HIV test.

CONCLUSIONS

The sequential emergence of assay reactivity allows the classification of primary HIV-<em>1</em> infection into distinct laboratory stages, which may facilitate the diagnosis of recent infection and stratification of patients enrolled in clinical trials. Quantitative analysis of preseroconversion replication rates of HIV is useful for projecting the yield and predictive value of assays targeting primary HIV infection.

The precise staging of hepatocellular carcinoma (HCC) based on the size and number of lesions that predict recurrence after orthotopic liver transplantation (OLT) has not been clearly established. We therefore analyzed the outcome of 70 consecutive patients with cirrhosis and HCC who underwent OLT over a <em>1</em>2-year period at our institution. Pathologic tumor staging of the explanted liver was based on the American Tumor Study Group modified Tumor-Node-Metastases (TNM) Staging Classification. Tumor recurrence occurred in <em>1</em><em>1</em>.4% of patients after OLT. The Kaplan-Meier survival rates at <em>1</em> and 5 years were 9<em>1</em>.3% and 72.4%, respectively, for patients with pT<em>1</em> or pT2 HCC; and 82.4% and 74.<em>1</em>%, respectively, for pT3 tumors (P =.87). Patients with pT4 tumors, however, had a significantly worse <em>1</em>-year survival of 33.3% (P =.000<em>1</em>). An alpha-fetoprotein (AFP) level>> <em>1</em>,000 ng/<em>mL</em>, total tumor diameter>> 8 cm, age>> or = 55 years and poorly differentiated histologic grade were also significant predictors for reduced survival in univariate analysis. Only pT4 stage and total tumor diameter remained statistically significant in multivariate analysis. Patients with HCC meeting the following criteria: solitary tumor < or = 6.5 cm, or < or = 3 nodules with the largest lesion < or = 4.5 cm and total tumor diameter < or = 8 cm, had survival rates of 90% and 75.2%, at <em>1</em> and 5 years, respectively, after OLT versus a 50% <em>1</em>-year survival for patients with tumors exceeding these limits (P =.0005). We conclude that the current criteria for OLT based on tumor size may be modestly expanded while still preserving excellent survival after OLT.

Ischemic preconditioning (Pre-con) is an adaptive response triggered by a brief ischemia applied before a prolonged coronary occlusion. We tested the hypothesis that repetitive ischemia applied during early reperfusion, i.e., postconditioning (Post-con), is cardio-protective by attenuating reperfusion injury. In anesthetized open-chest dogs, the left anterior descending artery (LAD) was occluded for 60 min and reperfused for 3 h. In controls (n = <em>1</em>0), there was no intervention. In Pre-con (n = 9), the LAD was occluded for 5 min and reperfused for <em>1</em>0 min before the prolonged occlusion. In Post-con (n = <em>1</em>0), at the start of reperfusion, three cycles of 30-s reperfusion and 30-s LAD reocclusion preceded the 3 h of reperfusion. Infarct size was significantly less in the Pre-con (<em>1</em>5 +/- 2%, P < 0.05) and Post-con (<em>1</em>4 +/- 2%, P < 0.05) groups compared with controls (25 +/- 3%). Tissue edema (% water content) in the area at risk was comparably reduced in Pre-con (78.3 +/- <em>1</em>.2, P < 0.05) and Post-con (79.7 +/- 0.6, P < 0.05) versus controls (8<em>1</em>.5 +/- 0.4). Polymorphonuclear neutrophil (PMN) accumulation (myeloperoxidase activity, Deltaabsorbance.min-<em>1</em>.g tissue-<em>1</em>) in the area at risk myocardium was comparably reduced in Post-con (<em>1</em>0.8 +/- 5.5, P < 0.05) and Pre-con (<em>1</em>3.4 +/- 3.4, P < 0.05) versus controls (47.4 +/- <em>1</em>5.3). Basal endothelial function measured by PMN adherence to postischemic LAD endothelium (PMNs/mm2) was comparably attenuated by Post-con and Pre-con (<em>1</em>5 +/- 0.6 and <em>1</em>2 +/- 0.6, P < 0.05) versus controls (37 +/- <em>1</em>.5), consistent with reduced expression of P-selectin on coronary vascular endothelium in Post-con and Pre-con. Endothelial function assessed by the maximal vasodilator response of postischemic LAD to acetylcholine was significantly greater in Post-con (<em>1</em>04 +/- 6%, P < 0.05) and Pre-con (<em>1</em>09 +/- 5%, P < 0.05) versus controls (7<em>1</em> +/- 8%). Plasma malondialdehyde (microM/<em>ml</em>), a product of lipid peroxidation, was significantly less at <em>1</em> h of reperfusion in Post-con (2.2 +/- 0.2, P < 0.05) versus controls (3.2 +/- 0.3) associated with a decrease in superoxide levels revealed by dihydroethidium staining in the myocardial area at risk. These data suggest that Post-con is as effective as Pre-con in reducing infarct size and preserving endothelial function. Post-con may be clinically applicable in coronary interventions, coronary artery bypass surgery, organ transplantation, and peripheral revascularization where reperfusion injury is expressed.

A unique cohort of HIV-<em>1</em>-infected long term nonprogressors (LTNP) with normal CD4(+) T cell counts and <50 copies/<em>ml</em> of plasma were prospectively recruited for study. HLA typing revealed a dramatic association between the HLA B*570<em>1</em> class I allele and nonprogressive infection [85% (<em>1</em><em>1</em> of <em>1</em>3) vs. 9.5% (<em>1</em>9 of 200) in progressors; P < 0. 00<em>1</em>]. Antigen-specific CD8(+) T cells were enumerated by flow cytometric detection of intracellular IFN-gamma in response to HIV antigens and HLA B*57-gag tetramer staining. No quantitative differences in the total HIV-specific CD8(+) T cell responses were observed between B*57(+) LTNP and five B*57(+) progressors (P = 0.4). Although similar frequencies of peptide specific CD8(+) T cells were also found, the gag-specific CD8(+) T cell response in the LTNP group was highly focused on peptides previously shown to be B*57-restricted. These findings indicate that, within this phenotypically and genotypically distinct cohort, a host immune factor is highly associated with restriction of virus replication and nonprogressive disease. They also strongly suggest a mechanism of virus specific immunity that directly operates through the B*570<em>1</em> molecule. Further characterization of qualitative differences in the virus-specific responses that distinguish HLA B*57(+) LTNP from progressors may ultimately define mechanisms of effective immune mediated restriction of virus replication.

A Klebsiella pneumoniae isolate showing moderate to high-level imipenem and meropenem resistance was investigated. The MICs of both drugs were <em>1</em>6 microg/<em>ml</em>. The beta-lactamase activity against imipenem and meropenem was inhibited in the presence of clavulanic acid. The strain was also resistant to extended-spectrum cephalosporins and aztreonam. Isoelectric focusing studies demonstrated three beta-lactamases, with pIs of 7.2 (SHV-29), 6.7 (KPC-<em>1</em>), and 5.4 (TEM-<em>1</em>). The presence of bla(SHV) and bla(TEM) genes was confirmed by specific PCRs and DNA sequence analysis. Transformation and conjugation studies with Escherichia coli showed that the beta-lactamase with a pI of 6.7, KPC-<em>1</em> (K. pneumoniae carbapenemase-<em>1</em>), was encoded on an approximately 50-kb nonconjugative plasmid. The gene, bla(KPC-<em>1</em>), was cloned in E. coli and shown to confer resistance to imipenem, meropenem, extended-spectrum cephalosporins, and aztreonam. The amino acid sequence of the novel carbapenem-hydrolyzing beta-lactamase, KPC-<em>1</em>, showed 45% identity to the pI 9.7 carbapenem-hydrolyzing beta-lactamase, Sme-<em>1</em>, from Serratia marcescens S6. Hydrolysis studies showed that purified KPC-<em>1</em> hydrolyzed not only carbapenems but also penicillins, cephalosporins, and monobactams. KPC-<em>1</em> had the highest affinity for meropenem. The kinetic studies also revealed that clavulanic acid and tazobactam inhibited KPC-<em>1</em>. An examination of the outer membrane proteins of the parent K. pneumoniae strain demonstrated that the strain does not express detectable levels of OmpK35 and OmpK37, although OmpK36 is present. We concluded that carbapenem resistance in K. pneumoniae strain <em>1</em>534 is mainly due to production of a novel Bush group 2f, class A, carbapenem-hydrolyzing beta-lactamase, KPC-<em>1</em>, although alterations in porin expression may also play a role.

Arachidonic acid is metabolised either by cyclooxygenases to produce prostaglandins and thromboxanes or by lipoxygenases to produce mono-, di- and trihydroxyeicosatetraenoic acids (HETEs). Polymorphonuclear leukocytes (PMNs) release HETEs, including mono- and dihydroxy fatty acids, when exposed to stimuli such as the calcium ionophore A23<em>1</em>87 (refs <em>1</em>, 2). The mono-HETEs are assumed to be of particular importance with respect to effects on leukocyte function because they have been shown to possess both chemotactic and chemokinetic activities towards PMNs and eosinophils. However, we have now shown that the chemokinetic and aggregating activities released from rat and human PMNs exposed to ionophore A23<em>1</em>87 (ref. 5) are not due to the release of mono-HETEs but to that of 5, <em>1</em>2-di-HETE (leukotriene B). This compound is active over the concentration range <em>1</em>0 pg <em>ml</em>-<em>1</em> to 5 ng <em>ml</em>-<em>1</em>.

BACKGROUND

Hypertension is a leading cause of end-stage renal disease (ESRD) in the United States, with no known treatment to prevent progressive declines leading to ESRD.

OBJECTIVE

To compare the effects of 2 levels of blood pressure (BP) control and 3 antihypertensive drug classes on glomerular filtration rate (GFR) decline in hypertension.

METHODS

Randomized 3 x 2 factorial trial with enrollment from February 1995 to September 1998.

METHODS

A total of 1094 African Americans aged 18 to 70 years with hypertensive renal disease (GFR, 20-65 mL/min per 1.73 m(2)) were recruited from 21 clinical centers throughout the United States and followed up for 3 to 6.4 years.

METHODS

Participants were randomly assigned to 1 of 2 mean arterial pressure goals, 102 to 107 mm Hg (usual; n = 554) or 92 mm Hg or less (lower; n = 540), and to initial treatment with either a beta-blocker (metoprolol 50-200 mg/d; n = 441), an angiotensin-converting enzyme inhibitor (ramipril 2.5-10 mg/d; n = 436) or a dihydropyridine calcium channel blocker, (amlodipine 5-10 mg/d; n = 217). Open-label agents were added to achieve the assigned BP goals.

METHODS

Rate of change in GFR (GFR slope); clinical composite outcome of reduction in GFR by 50% or more (or>> or =25 mL/min per 1.73 m2) from baseline, ESRD, or death. Three primary treatment comparisons were specified: lower vs usual BP goal; ramipril vs metoprolol; and amlodipine vs metoprolol.

RESULTS

Achieved BP averaged (SD) 128/78 (12/8) mm Hg in the lower BP group and 141/85 (12/7) mm Hg in the usual BP group. The mean (SE) GFR slope from baseline through 4 years did not differ significantly between the lower BP group (-2.21 [0.17] mL/min per 1.73 m2 per year) and the usual BP group (-1.95 [0.17] mL/min per 1.73 m2 per year; P =.24), and the lower BP goal did not significantly reduce the rate of the clinical composite outcome (risk reduction for lower BP group = 2%; 95% confidence interval [CI], -22% to 21%; P =.85). None of the drug group comparisons showed consistent significant differences in the GFR slope. However, compared with the metoprolol and amlodipine groups, the ramipril group manifested risk reductions in the clinical composite outcome of 22% (95% CI, 1%-38%; P =.04) and 38% (95% CI, 14%-56%; P =.004), respectively. There was no significant difference in the clinical composite outcome between the amlodipine and metoprolol groups.

CONCLUSIONS

No additional benefit of slowing progression of hypertensive nephrosclerosis was observed with the lower BP goal. Angiotensin-converting enzyme inhibitors appear to be more effective than beta-blockers or dihydropyridine calcium channel blockers in slowing GFR decline.

BACKGROUND

Whether correction of anemia in patients with stage 3 or 4 chronic kidney disease improves cardiovascular outcomes is not established.

METHODS

We rando<em>ml</em>y assigned 603 patients with an estimated glomerular filtration rate (GFR) of <em>1</em>5.0 to 35.0 <em>ml</em> per minute per <em>1</em>.73 m2 of body-surface area and mild-to-moderate anemia (hemoglobin level, <em>1</em><em>1</em>.0 to <em>1</em>2.5 g per deciliter) to a target hemoglobin value in the normal range (<em>1</em>3.0 to <em>1</em>5.0 g per deciliter, group <em>1</em>) or the subnormal range (<em>1</em>0.5 to <em>1</em><em>1</em>.5 g per deciliter, group 2). Subcutaneous erythropoietin (epoetin beta) was initiated at randomization (group <em>1</em>) or only after the hemoglobin level fell below <em>1</em>0.5 g per deciliter (group 2). The primary end point was a composite of eight cardiovascular events; secondary end points included left ventricular mass index, quality-of-life scores, and the progression of chronic kidney disease.

RESULTS

During the 3-year study, complete correction of anemia did not affect the likelihood of a first cardiovascular event (58 events in group <em>1</em> vs. 47 events in group 2; hazard ratio, 0.78; 95% confidence interval, 0.53 to <em>1</em>.<em>1</em>4; P=0.20). Left ventricular mass index remained stable in both groups. The mean estimated GFR was 24.9 <em>ml</em> per minute in group <em>1</em> and 24.2 <em>ml</em> per minute in group 2 at baseline and decreased by 3.6 and 3.<em>1</em> <em>ml</em> per minute per year, respectively (P=0.40). Dialysis was required in more patients in group <em>1</em> than in group 2 (<em>1</em>27 vs. <em>1</em><em>1</em><em>1</em>, P=0.03). General health and physical function improved significantly (P=0.003 and P<0.00<em>1</em>, respectively, in group <em>1</em>, as compared with group 2). There was no significant difference in the combined incidence of adverse events between the two groups, but hypertensive episodes and headaches were more prevalent in group <em>1</em>.

CONCLUSIONS

In patients with chronic kidney disease, early complete correction of anemia does not reduce the risk of cardiovascular events. (ClinicalTrials.gov number, NCT0032<em>1</em>9<em>1</em>9 [ClinicalTrials.gov].).

Before <em>1</em>985 at the Pitié-Salpêtrière Hospital in Paris (2,400 beds), resistance to cefotaxime in clinical isolates of Enterobacteriaceae involved only species producing inducible class <em>1</em> beta-lactamase. Between November <em>1</em>985 and April <em>1</em>987, however, 62 isolates (57 of Klebsiella pneumoniae and five of Escherichia coli) showed decreased susceptibility to cefotaxime (mean MIC, 8-<em>1</em>6 micrograms/<em>mL</em>). The transferability of cefotaxime resistance in E. coli K<em>1</em>2 was demonstrated for <em>1</em>5 of <em>1</em>6 selected isolates. By isoelectric focusing using iodometric detection with 20 mg of ceftriaxone/<em>1</em>00 <em>mL</em> and determination of substrate and inhibition profiles, three beta-lactamases mediating cefotaxime resistance were identified as SHV-2 (isoelectric point [pI] 7.6), CTX-<em>1</em> (pI 6.3), and "SHV-2-type" or SHV-3 (pI 6.98). The three beta-lactamases hydrolyzed penicillins and cephalosporins (including cefotaxime and ceftriaxone) and were therefore designated "extended broad-spectrum beta-lactamases" (EBS-Bla). The enzymes conferred to derivatives a high level of resistance to amoxicillin, ticarcillin, piperacillin, and cephalothin and a decreased degree of susceptibility (i.e., MICs increased by <em>1</em>0- to 800-fold) to cefotaxime, ceftriaxone, ceftazidime, and aztreonam. These beta-lactamases did not affect the activity of cephamycins (cefoxitin, cefotetan, moxalactam) or imipenem. Synergy between clavulanate or sulbactam (2 micrograms/<em>mL</em>) and amoxicillin was greater against derivatives producing EBS-Bla than against those producing TEM-<em>1</em>, TEM-2, or SHV-<em>1</em>; this synergy was greater with clavulanate than with sulbactam against derivatives producing SHV-2 and the SHV-2-type enzyme but was similar with clavulanate and sulbactam against those producing CTX-<em>1</em>. A double-disk synergy test performed with cefotaxime and Augmentin disks (placed 30 mm apart, center to center) seemed a useful and specific test for the detection of strains producing EBS-Bla.

The determination and characterization of a cannabinoid receptor from brain are reported. A biologically active bicyclic cannabinoid analgetic CP-55,940 was tritium-labeled to high specific activity. Conditions for binding to rat brain P2 membranes and synaptosomes were established. The pH optimum was between 7 and 8, and specific binding could be eliminated by heating the membranes to 60 degrees. Binding to the P2 membranes was linear within the range of <em>1</em>0 to 50 micrograms of protein/<em>ml</em>. Specific binding (defined as total binding displaced by <em>1</em> microM delta 9-tetrahydrocannabinol (delta 9-THC) or <em>1</em>00 nM desacetyllevonantradol) was saturable. The Kd determined from Scatchard analysis was <em>1</em>33 pM, and the Bmax for rat cortical P2 membranes was <em>1</em>.85 pmol/mg of protein. The Hill coefficient for [3H]CP-55,940 approximated <em>1</em>, indicating that, under the conditions of assay, a single class of binding sites was determined that did not exhibit cooperativity. The binding was rapid (kon approximately 2.6 x <em>1</em>0(-4) pM-<em>1</em> min-<em>1</em>) and reversible (Koff approximately 0.0<em>1</em>6 min-<em>1</em>) and (koff' greater than 0.06 min-<em>1</em>). The two Kd values estimated from the kinetic constants approximately 55 pM and exceeded 200 pM, respectively. The binding of the agonist ligand [3H]CP-55,940 was decreased by the nonhydrolyzable GTP analog guanylylimidodiphosphate. The guanine nucleotide induced a more rapid dissociation of the ligand from the binding site, consistent with an allosteric regulation of the putative receptor by a G protein. The binding was also sensitive to MgCl2 and CaCl2. Binding of [3H]CP-55,940 was displaced by cannabinoid drugs in the following order of potency: CP-55,940 greater than or equal to desacetyllevonantradol greater than <em>1</em><em>1</em>-OH-delta 9-THC = delta 9-THC greater than cannabinol. Cannabidiol and cannabigerol displaced [3H]CP-55,940 by less than 50% at <em>1</em> microM concentrations. The (-)-isomer of CP-55,940 displaced with 50-fold greater potency than the (+)-isomer. This pharmacology is comparable to both the inhibition of adenylate cyclase in vitro and the analgetic activity of these compounds in vivo. The criteria for a high affinity, stereoselective, pharmacologically distinct cannabinoid receptor in brain tissue have been fulfilled.

BACKGROUND

Mobile (cell) phone communication has been suggested as a method to improve delivery of health services. However, data on the effects of mobile health technology on patient outcomes in resource-limited settings are limited. We aimed to assess whether mobile phone communication between health-care workers and patients starting antiretroviral therapy in Kenya improved drug adherence and suppression of plasma HIV-<em>1</em> RNA load.

METHODS

WelTel Kenya<em>1</em> was a multisite randomised clinical trial of HIV-infected adults initiating antiretroviral therapy (ART) in three clinics in Kenya. Patients were randomised (<em>1</em>:<em>1</em>) by simple randomisation with a random number generating program to a mobile phone short message service (SMS) intervention or standard care. Patients in the intervention group received weekly SMS messages from a clinic nurse and were required to respond within 48 h. Randomisation, laboratory assays, and analyses were done by investigators masked to treatment allocation; however, study participants and clinic staff were not masked to treatment. Primary outcomes were self-reported ART adherence (>95% of prescribed doses in the past 30 days at both 6 and <em>1</em>2 month follow-up visits) and plasma HIV-<em>1</em> viral RNA load suppression (<400 copies per mL) at <em>1</em>2 months. The primary analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, NCT00830622.

RESULTS

Between May, 2007, and October, 2008, we randomly assigned 538 participants to the SMS intervention (n=273) or to standard care (n=265). Adherence to ART was reported in <em>1</em>68 of 273 patients receiving the SMS intervention compared with <em>1</em>32 of 265 in the control group (relative risk [RR] for non-adherence 0·8<em>1</em>, 95% CI 0·69-0·94; p=0·006). Suppressed viral loads were reported in <em>1</em>56 of 273 patients in the SMS group and <em>1</em>28 of 265 in the control group, (RR for virologic failure 0·84, 95% CI 0·7<em>1</em>-0·99; p=0·04). The number needed to treat (NNT) to achieve greater than 95% adherence was nine (95% CI 5·0-29·5) and the NNT to achieve viral load suppression was <em>1</em><em>1</em> (5·8-227·3).

CONCLUSIONS

Patients who received SMS support had significantly improved ART adherence and rates of viral suppression compared with the control individuals. Mobile phones might be effective tools to improve patient outcome in resource-limited settings.

BACKGROUND

US President's Emergency Plan for AIDS Relief.

Despite extensive laboratory investigations in patients with respiratory tract infections, no microbiological cause can be identified in a significant proportion of patients. In the past 3 years, several novel respiratory viruses, including human metapneumovirus, severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV), and human coronavirus NL63, were discovered. Here we report the discovery of another novel coronavirus, coronavirus HKU<em>1</em> (CoV-HKU<em>1</em>), from a 7<em>1</em>-year-old man with pneumonia who had just returned from Shenzhen, China. Quantitative reverse transcription-PCR showed that the amount of CoV-HKU<em>1</em> RNA was 8.5 to 9.6 x <em>1</em>0(6) copies per <em>ml</em> in his nasopharyngeal aspirates (NPAs) during the first week of the illness and dropped progressively to undetectable levels in subsequent weeks. He developed increasing serum levels of specific antibodies against the recombinant nucleocapsid protein of CoV-HKU<em>1</em>, with immunoglobulin M (IgM) titers of <em>1</em>:20, <em>1</em>:40, and <em>1</em>:80 and IgG titers of (<em>1</em>:<em>1</em>,000, <em>1</em>:2,000, and <em>1</em>:8,000 in the first, second and fourth weeks of the illness, respectively. Isolation of the virus by using various cell lines, mixed neuron-glia culture, and intracerebral inoculation of suckling mice was unsuccessful. The complete genome sequence of CoV-HKU<em>1</em> is a 29,926-nucleotide, polyadenylated RNA, with G+C content of 32%, the lowest among all known coronaviruses with available genome sequence. Phylogenetic analysis reveals that CoV-HKU<em>1</em> is a new group 2 coronavirus. Screening of 400 NPAs, negative for SARS-CoV, from patients with respiratory illness during the SARS period identified the presence of CoV-HKU<em>1</em> RNA in an additional specimen, with a viral load of <em>1</em>.<em>1</em>3 x <em>1</em>0(6) copies per <em>ml</em>, from a 35-year-old woman with pneumonia. Our data support the existence of a novel group 2 coronavirus associated with pneumonia in humans.

BACKGROUND

The prevalence of chronic kidney disease is high in developing countries. However, no national survey of chronic kidney disease has been done incorporating both estimated glomerular filtration rate (eGFR) and albuminuria in a developing country with the economic diversity of China. We aimed to measure the prevalence of chronic kidney disease in China with such a survey.

METHODS

We did a cross-sectional survey of a nationally representative sample of Chinese adults. Chronic kidney disease was defined as eGFR less than 60 <em>mL</em>/min per <em>1</em>·73 m(2) or the presence of albuminuria. Participants completed a lifestyle and medical history questionnaire and had their blood pressure measured, and blood and urine samples taken. Serum creatinine was measured and used to estimate glomerular filtration rate. Urinary albumin and creatinine were tested to assess albuminuria. The crude and adjusted prevalence of indicators of kidney damage were calculated and factors associated with the presence of chronic kidney disease analysed by logistic regression.

RESULTS

50,550 people were invited to participate, of whom 47,204 agreed. The adjusted prevalence of eGFR less than 60 <em>mL</em>/min per <em>1</em>·73 m(2) was <em>1</em>·7% (95% CI <em>1</em>·5-<em>1</em>·9) and of albuminuria was 9·4% (8·9-<em>1</em>0·0). The overall prevalence of chronic kidney disease was <em>1</em>0·8% (<em>1</em>0·2-<em>1</em><em>1</em>·3); therefore the number of patients with chronic kidney disease in China is estimated to be about <em>1</em><em>1</em>9·5 million (<em>1</em><em>1</em>2·9-<em>1</em>25·0 million). In rural areas, economic development was independently associated with the presence of albuminuria. The prevalence of chronic kidney disease was high in north (<em>1</em>6·9% [<em>1</em>5·<em>1</em>-<em>1</em>8·7]) and southwest (<em>1</em>8·3% [<em>1</em>6·4-20·4]) regions compared with other regions. Other factors independently associated with kidney damage were age, sex, hypertension, diabetes, history of cardiovascular disease, hyperuricaemia, area of residence, and economic status.

CONCLUSIONS

Chronic kidney disease has become an important public health problem in China. Special attention should be paid to residents in economically improving rural areas and specific geographical regions in China.

BACKGROUND

The Ministry of Science and Technology (China); the Science and Technology Commission of Shanghai; the National Natural Science Foundation of China; the Department of Health, Jiangsu Province; the Sichuan Science and Technology Department; the Ministry of Education (China); the International Society of Nephrology Research Committee; and the China Health and Medical Development Foundation.

This large-volume, single-institution review examines factors influencing long-term survival after resection in patients with adenocarcinoma of the head, neck, uncinate process, body, or tail of the pancreas. Between January <em>1</em>984 and July <em>1</em>999 inclusive, 6<em>1</em>6 patients with adenocarcinoma of the pancreas underwent surgical resection. A retrospective analysis of a prospectively collected database was performed. Both univariate and multivariate models were used to determine the factors influencing survival. Of the 6<em>1</em>6 patients, 526 (85%) underwent pancreaticoduodenectomy for adenocarcinoma of the head, neck, or uncinate process of the pancreas, 52 (9%) underwent distal pancreatectomy for adenocarcinoma of the body or tail, and 38 (6%) underwent total pancreatectomy for adenocarcinoma extensively involving the gland. The mean age of the patients was 64.3 years, with 54% being male and 9<em>1</em>% being white. The overall perioperative mortality rate was 2.3%, whereas the incidence of postoperative complications was 30%. The median postoperative length of stay was <em>1</em><em>1</em> days. The mean tumor diameter was 3.2 cm, with 72% of patients having positive lymph nodes, 30% having positive resection margins, and 36% having poorly differentiated tumors. Patients undergoing distal pancreatectomy for left-sided lesions had larger tumors (4.7 vs. 3.<em>1</em> cm, P < 0.000<em>1</em>), but fewer node-positive resections (59% vs. 73%, P = 0.03) and fewer poorly differentiated tumors (29% vs. 36%, P < 0.00<em>1</em>), as compared to those undergoing pancreaticoduodenectomy for right-sided lesions. The overall survival of the entire cohort was 63% at <em>1</em> year and <em>1</em>7% at 5 years, with a median survival of <em>1</em>7 months. For right-sided lesions the <em>1</em>- and 5-year survival rates were 64% and <em>1</em>7%, respectively, compared to 50% and <em>1</em>5% for left-sided lesions. Factors shown to have favorable independent prognostic significance by multivariate analysis were negative resection margins (hazard ratio [HR] = 0.64, confidence interval [CI] = 0.50 to 0.82, P = 0.0004), tumor diameter less than 3 cm (HR = 0.72, CI = 0.57 to 0.90, P = 0.004), estimated blood loss less than 750 <em>ml</em> (HR = 0.75, CI = 0.58 to 0.96, P = 0.02), well/moderate tumor differentiation (HR = 0.7<em>1</em>, CI = 0.56 to 0.90, P = 0.005), and postoperative chemoradiation (HR = 0.50, CI = 0.39 to 0.64, P < 0.000<em>1</em>). Tumor location in head, neck, or uncinate process approached significance in the final multivariate model (HR = 0.60, CI = 0.35 to <em>1</em>.0, P = 0.06). Pancreatic resection remains the only hope for long-term survival in patients with adenocarcinoma of the pancreas. Completeness of resection and tumor characteristics including tumor size and degree of differentiation are important independent prognostic indicators. Adjuvant chemoradiation is a strong predictor of outcome and likely decreases the independent significance of tumor location and nodal status.

Monoclonal antibody GK<em>1</em>.5 recognizes a previously undescribed murine T cell surface molecule, designated L3T4, which migrates on SDS-PAGE under reducing conditions as a single band with an apparent m.w. of 52,000. L3T4 is expressed by approximately 80% of thymocytes and by approximately 20% of spleen cells. There appears to be poor correlation between expression of L3T4 by functional T cell clones and expression of Lyt-2, expression of the cytolytic phenotype, and class I MHC antigen reactivity. On the other hand, both a class II MHC antigen-reactive HTL clone and an Lyt-<em>1</em>- <em>Mls</em>-reactive HTL clone express L3T4. Analysis of the effect of mAb GK<em>1</em>.5 on PFC responses in adoptive transfer suggests that L3T4 is expressed by the helper/inducer subset of murine T cells. Expression of L3T4 by murine T cells, however, may correlate primarily with class II MHC antigen reactivity rather than with functional phenotype; mAb GK<em>1</em>.5 profoundly blocks antigen-specific cytolysis by the cloned class II MHC antigen-reactive CTL line A<em>1</em>5-<em>1</em>.<em>1</em>7. Antigen-specific cytolysis by A<em>1</em>5-<em>1</em>.<em>1</em>7 is blocked by mAb GK<em>1</em>.5 at a step before the lethal hit. Collectively, the flow cytometric, functional, and biochemical data indicate that L3T4 is similar to the human Leu-3/T4 molecule.

We have previously examined the specificities of 28 commercially available compounds, reported to be relatively selective inhibitors of particular serine/threonine-specific protein kinases [Davies, Reddy, Caivano and Cohen (2000) Biochem. J. 35<em>1</em>, 95-<em>1</em>05]. In the present study, we have extended this analysis to a further <em>1</em>4 compounds. Of these, indirubin-3'-monoxime, SP 600<em>1</em>25, KT 5823 and <em>ML</em>-9 were found to inhibit a number of protein kinases and conclusions drawn from their use in cell-based assays are likely to be erroneous. Kenpaullone, Alsterpaullone, Purvalanol, Roscovitine, pyrazolopyrimidine <em>1</em> (PP<em>1</em>), PP2 and <em>ML</em>-7 were more specific, but still inhibited two or more protein kinases with similar potency. Our results suggest that the combined use of Roscovitine and Kenpaullone may be useful for identifying substrates and physiological roles of cyclin-dependent protein kinases, whereas the combined use of Kenpaullone and LiCl may be useful for identifying substrates and physiological roles of glycogen synthase kinase 3. The combined use of SU 6656 and either PP<em>1</em> or PP2 may be useful for identifying substrates of Src family members. Epigallocatechin 3-gallate, one of the main polyphenolic constituents of tea, inhibited two of the 28 protein kinases in the panel, dual-specificity, tyrosine-phosphorylated and regulated kinase <em>1</em>A (DYRK<em>1</em>A; IC(50)=0.33 microM) and p38-regulated/activated kinase (PRAK; IC(50)=<em>1</em>.0 microM).

BACKGROUND

Severe sepsis is typically characterized by initial cytokine-mediated hyperinflammation. Whether this hyperinflammatory phase is followed by immunosuppression is controversial. Animal studies suggest that multiple immune defects occur in sepsis, but data from humans remain conflicting.

OBJECTIVE

To determine the association of sepsis with changes in host innate and adaptive immunity and to examine potential mechanisms for putative immunosuppression.

METHODS

Rapid postmortem spleen and lung tissue harvest was performed at the bedsides of 40 patients who died in intensive care units (ICUs) of academic medical centers with active severe sepsis to characterize their immune status at the time of death (2009-20<em>1</em><em>1</em>). Control spleens (n = 29) were obtained from patients who were declared brain-dead or had emergent splenectomy due to trauma; control lungs (n = 20) were obtained from transplant donors or from lung cancer resections.

METHODS

Cytokine secretion assays and immunophenotyping of cell surface receptor-ligand expression profiles were performed to identify potential mechanisms of immune dysfunction. Immunohistochemical staining was performed to evaluate the loss of immune effector cells.

RESULTS

The mean ages of patients with sepsis and controls were 7<em>1</em>.7 (SD, <em>1</em>5.9) and 52.7 (SD, <em>1</em>5.0) years, respectively. The median number of ICU days for patients with sepsis was 8 (range, <em>1</em>-<em>1</em>95 days), while control patients were in ICUs for 4 or fewer days. The median duration of sepsis was 4 days (range, <em>1</em>-40 days). Compared with controls, anti-CD3/anti-CD28-stimulated splenocytes from sepsis patients had significant reductions in cytokine secretion at 5 hours: tumor necrosis factor, 536<em>1</em> (95% CI, 3327-7485) pg/<em>mL</em> vs 4<em>1</em>8 (95% CI, 98-738) pg/<em>mL</em>; interferon γ, <em>1</em>374 (95% CI, 550-2<em>1</em>97) pg/<em>mL</em> vs 37.5 (95% CI, -5 to 80) pg/<em>mL</em>; interleukin 6, 369<em>1</em> (95% CI, 23<em>1</em>3-5070) vs 365 (95% CI, 87-642) pg/<em>mL</em>; and interleukin <em>1</em>0, 633 (95% CI, -269 to <em>1</em>534) vs 58 (95% CI, -39 to <em>1</em>56) pg/<em>mL</em>; (P < .00<em>1</em> for all). There were similar reductions in 5-hour lipopolysaccharide-stimulated cytokine secretion. Cytokine secretion in sepsis patients was generally less than <em>1</em>0% that in controls, independent of age, duration of sepsis, corticosteroid use, and nutritional status. Although differences existed between spleen and lung, flow cytometric analysis showed increased expression of selected inhibitory receptors and ligands and expansion of suppressor cell populations in both organs. Unique differences in cellular inhibitory molecule expression existed in immune cells isolated from lungs of sepsis patients vs cancer patients and vs transplant donors. Immunohistochemical staining showed extensive depletion of splenic CD4, CD8, and HLA-DR cells and expression of ligands for inhibitory receptors on lung epithelial cells.

CONCLUSIONS

Patients who die in the ICU following sepsis compared with patients who die of nonsepsis etiologies have biochemical, flow cytometric, and immunohistochemical findings consistent with immunosuppression. Targeted immune-enhancing therapy may be a valid approach in selected patients with sepsis.

Two receptor systems for corticosterone (CORT) can be distinguished in rat brain: mineralocorticoid-like or CORT receptors (CR) and glucocorticoid receptors (GR). The microdistribution and extent of occupation of each receptor population by CORT were studied. The CR system is restricted predominantly to the lateral septum and hippocampus. Within the hippocampus, the highest density occurs in the subiculum +/- CA<em>1</em> cell field (<em>1</em>44 fmol/mg protein) and the dentate gyrus (<em>1</em>04 fmol/mg protein). Affinity of CR for CORT was very high (Kd, approximately 0.5 nM). The GR system has a more widespread distribution in the brain. The highest density for GR is in the lateral septum (<em>1</em>95 fmol/mg protein), the dentate gyrus (<em>1</em>33 fmol/mg protein), the nucleus tractus solitarii and central amygdala. Substantial amounts of GR are present in the paraventricular nucleus and locus coeruleus and low amounts in the raphe area and the subiculum + CA<em>1</em> cell field. The affinity of GR for CORT (Kd, approximately 2.5-5 nM) was 6- to <em>1</em>0-fold lower than that of CR. Occupation of CR by endogenous ligand was 89.5% during morning trough levels of pituitary-adrenal activity (plasma CORT, <em>1</em>.4 micrograms/<em>1</em>00 <em>ml</em>). Similar levels of occupation (88.7% and 97.6%) were observed at the diurnal peak (plasma CORT, 27 micrograms/<em>1</em>00 <em>ml</em>) and after <em>1</em> h of restraint stress (plasma CORT, 25 micrograms/<em>1</em>00 <em>ml</em>), respectively. Furthermore, a dose of <em>1</em> microgram CORT/<em>1</em>00 g BW, sc, resulted in 80% CORT receptor occupation, whereas GR were not occupied. For 50% occupation of GR, doses needed to be increased to 50-<em>1</em>00 micrograms/<em>1</em>00 g BW, and for 95% occupation, a dose of <em>1</em> mg CORT was required. The plasma CORT level at the time of half-maximal GR occupation was about 25 micrograms/<em>1</em>00 <em>ml</em>, which is in the range of levels attained after stress or during the diurnal peak of pituitary-adrenal activity. Thus, CR are extensively filled (greater than 90%) with endogenous CORT under most circumstances, while GR become occupied concurrent with increasing plasma CORT concentrations due to stress or diurnal rhythm. We conclude that CORT action via CR may be involved in a tonic (permissive) influence on brain function with the septohippocampal complex as a primary target. In view of the almost complete occupation of CR by endogenous hormones, the regulation of the CORT signal via CR will, most likely, be by alterations in the number of such receptors. In contrast, CORT action via GR is involved in its feedback action on stress-activated brain mechanisms, and GR occur widely in the brain.(ABSTRACT TRUNCATED AT 400 WORDS)

In December 20<em>1</em>9, a coronavirus 20<em>1</em>9 (COVID-<em>1</em>9) disease outbreak occurred in Wuhan, Hubei Province, China, and rapidly spread to other areas worldwide. Although diffuse alveolar damage and acute respiratory failure were the main features, the involvement of other organs needs to be explored. Since information on kidney disease in patients with COVID-<em>1</em>9 is limited, we determined the prevalence of acute kidney injury (AKI) in patients with COVID-<em>1</em>9. Further, we evaluated the association between markers of abnormal kidney function and death in patients with COVID-<em>1</em>9. This was a prospective cohort study of 70<em>1</em> patients with COVID-<em>1</em>9 admitted in a tertiary teaching hospital that also encompassed three affiliates following this major outbreak in Wuhan in 2020 of whom <em>1</em><em>1</em>3 (<em>1</em>6.<em>1</em>%) died in hospital. Median age of the patients was 63 years (interquartile range, 50-7<em>1</em>), including 367 men and 334 women. On admission, 43.9% of patients had proteinuria and 26.7% had hematuria. The prevalence of elevated serum creatinine, elevated blood urea nitrogen and estimated glomerular filtration under 60 <em>ml</em>/min/<em>1</em>.73m² were <em>1</em>4.4, <em>1</em>3.<em>1</em> and <em>1</em>3.<em>1</em>%, respectively. During the study period, AKI occurred in 5.<em>1</em>% patients. Kaplan-Meier analysis demonstrated that patients with kidney disease had a significantly higher risk for in-hospital death. Cox proportional hazard regression confirmed that elevated baseline serum creatinine (hazard ratio: 2.<em>1</em>0, 95% confidence interval: <em>1</em>.36-3.26), elevated baseline blood urea nitrogen (3.97, 2.57-6.<em>1</em>4), AKI stage <em>1</em> (<em>1</em>.90, 0.76-4.76), stage 2 (3.5<em>1</em>, <em>1</em>.49-8.26), stage 3 (4.38, 2.3<em>1</em>-8.3<em>1</em>), proteinuria <em>1</em>+ (<em>1</em>.80, 0.8<em>1</em>-4.00), 2+∼3+ (4.84, 2.00-<em>1</em><em>1</em>.70), and hematuria <em>1</em>+ (2.99, <em>1</em>.39-6.42), 2+∼3+ (5.56,2.58- <em>1</em>2.0<em>1</em>) were independent risk factors for in-hospital death after adjusting for age, sex, disease severity, comorbidity and leukocyte count. Thus, our findings show the prevalence of kidney disease on admission and the development of AKI during hospitalization in patients with COVID-<em>1</em>9 is high and is associated with in-hospital mortality. Hence, clinicians should increase their awareness of kidney disease in patients with severe COVID-<em>1</em>9.

A highly sensitive and selective high-performance liquid chromatographic method with short-wavelength UV detection is described for the determination of ionic compounds in biological fluids, which was applied to two basic compounds, 2-(3-[4-(4-fluorophenyl)-<em>1</em>-piperazinyl]propyl)-6,7,8,9-tetrahydro-2H-nap htho [2,3-b][<em>1</em>,4]oxazin-3(4H)-one (I) and methyl 2-(4-diphenylmethyl-<em>1</em>-piperazinyl)ethyl (+/-)-<em>1</em>,4-dihydro-2,6-dimethyl-4-(m-nitrophenyl)-3,5-pyridinedicarboxyla te (II), in human serum. The method is based on the combination of the column-switching technique and ion-pair chromatography. In the first ODS column, the analyte is pre-separated from endogenous substances in serum by ion-pair chromatography. After column switching, in the second ODS column the heart-cut fraction containing the analyte is further separated by non-ion-pair chromatography from coeluted endogenous substances from the first ODS column. The proposed method offered high sensitivity and selectivity with UV detection at 2<em>1</em>5 nm for I and 230 nm for II. The detection limits were 0.2 ng/<em>ml</em> for both I and II using <em>1</em> <em>ml</em> of serum. The principle of the proposed method would be applicable to both acidic and basic compounds in biological fluids with a suitable ion-pair reagent.

<b>Background:</b> The outbreak of coronavirus disease 20<em>1</em>9 (COVID-<em>1</em>9) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has posed great threat to human health. T cells play a critical role in antiviral immunity but their numbers and functional state in COVID-<em>1</em>9 patients remain largely unclear. <b>Methods:</b> We retrospectively reviewed the counts of T cells and serum cytokine concentration from data of 522 patients with laboratory-confirmed COVID-<em>1</em>9 and 40 healthy controls. In addition, the expression of T cell exhaustion markers were measured in <em>1</em>4 COVID-<em>1</em>9 cases. <b>Results:</b> The number of total T cells, CD4⁺ and CD8⁺ T cells were dramatically reduced in COVID-<em>1</em>9 patients, especially in patients requiring Intensive Care Unit (ICU) care. Counts of total T cells, CD8⁺ T cells or CD4⁺ T cells lower than 800, 300, or 400/<em>μL</em>, respectively, were negatively correlated with patient survival. T cell numbers were negatively correlated to serum IL-6, IL-<em>1</em>0, and TNF-α concentration, with patients in the disease resolution period showing reduced IL-6, IL-<em>1</em>0, and TNF-α concentrations and restored T cell counts. T cells from COVID-<em>1</em>9 patients had significantly higher levels of the exhausted marker PD-<em>1</em>. Increasing PD-<em>1</em> and Tim-3 expression on T cells was seen as patients progressed from prodromal to overtly symptomatic stages. <b>Conclusions:</b> T cell counts are reduced significantly in COVID-<em>1</em>9 patients, and the surviving T cells appear functionally exhausted. Non-ICU patients with total T cells counts lower than 800/<em>μL</em> may still require urgent intervention, even in the immediate absence of more severe symptoms due to a high risk for further deterioration in condition.

<strong class="sub-title">Keywords:</strong> COVID-<em>1</em>9; SARS- CoV-2; T cell exhaustion; T cell reduction; cytokine strom.

A growth factor for vascular endothelial cells was identified in the media conditioned by bovine pituitary follicular cells and purified to homogeneity by a combination of ammonium sulfate precipitation, heparin-sepharose affinity chromatography and two reversed phase HPLC steps. The growth factor was a cationic, heat stable and relatively acid stable protein and had a molecular weight, as assessed by silver-stained SDS-PAGE gel, of approximately 45,000 under non reducing conditions and approximately 23,000 under reducing conditions. The purified growth factor had a maximal mitogenic effect on adrenal cortex-derived capillary endothelial cells at the concentration of <em>1</em>-<em>1</em>.2 ng/<em>ml</em> (22-26 pM). Further characterization of the bioactivity of the growth factor reveals that it exerts mitogenic effects also on vascular endothelial cells isolated from several districts but not on adrenal cortex cells, lens epithelial cells, corneal endothelial cells, keratynocytes or BHK-2<em>1</em> fibroblasts, indicating that its target cells specificity is unlike that of any previously characterized growth factor. Microsequencing reveals a unique N-terminal amino acid sequence. On the basis of its apparent target cell selectivity, we propose to name this factor vascular endothelial growth factor (VEGF).

BACKGROUND

The probability of HIV-<em>1</em> transmission per coital act in representative African populations is unknown. We aimed to calculate this probability overall, and to estimate how it is affected by various factors thought to influence infectivity.

METHODS

<em>1</em>74 monogamous couples, in which one partner was HIV-<em>1</em> positive, were retrospectively identified from a population cohort in Rakai, Uganda. Frequency of intercourse and reliability of reporting within couples was assessed prospectively. HIV-<em>1</em> seroconversion was determined in the uninfected partners, and HIV-<em>1</em> viral load was measured in the infected partners. Adjusted rate ratios of transmission per coital act were estimated by Poisson regression. Probabilities of transmission per act were estimated by log-log binomial regression for quartiles of age and HIV-<em>1</em> viral load, and for symptoms or diagnoses of sexually transmitted diseases (STDs) in the HIV-<em>1</em>-infected partners.

RESULTS

The mean frequency of intercourse was 8.9 per month, which declined with age and HIV-<em>1</em> viral load. Members of couples reported similar frequencies of intercourse. The overall unadjusted probability of HIV-<em>1</em> transmission per coital act was 0.00<em>1</em><em>1</em> (95% CI 0.0008-0.00<em>1</em>5). Transmission probabilities increased from 0.000<em>1</em> per act at viral loads of less than <em>1</em>700 copies/mL to 0.0023 per act at 38 500 copies/mL or more (p=0.002), and were 0.004<em>1</em> with genital ulceration versus 0.00<em>1</em><em>1</em> without (p=0.02). Transmission probabilities per act did not differ significantly by HIV-<em>1</em> subtypes A and D, sex, STDs, or symptoms of discharge or dysuria in the HIV-<em>1</em>-positive partner.

CONCLUSIONS

Higher viral load and genital ulceration are the main determinants of HIV-<em>1</em> transmission per coital act in this Ugandan population.

Hepatocyte growth factor (HGF) is the most potent mitogen for mature parenchymal hepatocytes in primary culture, and seems to be a hepatotrophic factor that acts as a trigger for liver regeneration after partial hepatectomy and liver injury. The partial purification and characterization of HGF have been reported. We have demonstrated that pure HGF from rat platelets is a new growth factor effective at concentrations as low as <em>1</em> ng <em>ml</em>-<em>1</em>. The effects of HGF and epidermal growth factor (EGF) are additive. The activity of HGF is not species-specific, although it does not stimulate growth in Swiss 3T3 fibroblasts. HGF has a relative molecular mass (Mr) of 82,000 and is a heterodimer composed of a large alpha-subunit of Mr 69,000 and a small beta-subunit of Mr 34,000. Here we report the amino-acid sequence of human HGF determined by complementary DNA cloning and the expression of biologically active human HGF from COS-<em>1</em> cells transfected with cloned cDNA. The nucleotide sequence of the human HGF cDNA reveals that both alpha- and beta-chains are contained in a single open reading frame coding for a pre-pro precursor protein of 728 amino acids.