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2.
J Virol ; 75(18): 8538-46, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11507199

ABSTRACT

The helper component of Cauliflower mosaic virus is encoded by viral gene II. This protein (P2) is dispensable for virus replication but required for aphid transmission. The purification of P2 has never been reported, and hence its biochemical properties are largely unknown. We produced the P2 protein via a recombinant baculovirus with a His tag fused at the N terminus. The fusion protein was purified by affinity chromatography in a soluble and biologically active form. Matrix-assisted laser desorption time-of-flight mass spectrometry demonstrated that P2 is not posttranslationally modified. UV circular dichroism revealed the secondary structure of P2 to be 23% alpha-helical. Most alpha-helices are suggested to be located in the C-terminal domain. Using size exclusion chromatography and aphid transmission testing, we established that the active form of P2 assembles as a huge soluble oligomer containing 200 to 300 subunits. We further showed that P2 can also polymerize as long paracrystalline filaments. We mapped P2 domains involved in P2 self-interaction, presumably through coiled-coil structures, one of which is proposed to form a parallel trimer. These regions have previously been reported to also interact with viral P3, another protein involved in aphid transmission. Possible interference between the two types of interaction is discussed with regard to the biological activity of P2.


Subject(s)
Caulimovirus/chemistry , Viral Proteins/chemistry , Amino Acid Sequence , Animals , Binding Sites , Cell Line , Molecular Sequence Data , Oligopeptides/chemistry , Oligopeptides/genetics , Oligopeptides/metabolism , Polymers , Protein Processing, Post-Translational , Protein Structure, Secondary , Spodoptera , Viral Proteins/genetics , Viral Proteins/metabolism
3.
Am J Med ; 111(2): 120-5, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11498065

ABSTRACT

PURPOSE: We examined the outcomes of bloodstream infection in men and in women and whether any sex-related differences were explained by underlying disorders, severity of disease, or clinical management. SUBJECTS AND METHODS: Using a prospectively collected database, we compared in-hospital mortality in men and women. We used multivariable logistic regression analysis to test whether sex-related differences could be due to potential confounders. RESULTS: Of 4250 patients with bloodstream infections, 1750 (41%) had hospital-acquired infections. The overall case fatality was 31% (625 of 2032) in women and 29% (631 of 2218, P = 0.1) in men. However, 43% (325/758) of the women with hospital-acquired infections died, compared with 33% (327/992) of the men (P = 0.0001). In a multivariate analysis, female sex was associated with greater mortality in patients with hospital-acquired infections (odds ratio = 1.7; 95% confidence interval: 1.1 to 2.6). The excess mortality in women was mainly seen in patients with major underlying disorders (fatality rate of 45% [234 of 525] in women vs. 32% in men [234 of 743, P = 0.0001). CONCLUSIONS: Mortality in women with hospital-acquired bloodstream infections is substantially greater than in men. The excess mortality was concentrated in women with severe underlying disorders, suggesting that sepsis might have accentuated differences in the outcome of underlying disorders in women.


Subject(s)
Cross Infection/mortality , Sepsis/mortality , Adult , Aged , Analysis of Variance , Confounding Factors, Epidemiologic , Cross Infection/etiology , Databases, Factual , Female , Hospital Mortality , Humans , Logistic Models , Middle Aged , Odds Ratio , Prospective Studies , Risk Factors , Sepsis/etiology , Severity of Illness Index , Sex Distribution
4.
Ren Fail ; 22(1): 99-108, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10718286

ABSTRACT

Pre-existing renal insufficiency serves as a common risk factor in the development of acute renal failure. Acute renal failure is a common finding in patients with bacteremia and is associated with poor prognosis. A total of 2722 consecutive patients 18 years old or older, fulfilling strike criteria of bacteremia or fungemia were prospectively evaluated to establish the prognostic importance of pre-existing renal insufficiency in bacteremic patients. They were classified according to serum creatinine levels upon admission into three groups. 915 patients had normal creatinine levels (< or = 1.0 mg/dL), 1528 had mild to moderate renal failure (creatinine 1.1-3 mg/dL) and 279 patients had severe renal failure upon admission (creatinine > 3.0 mg/dL). Mild to severe renal failure upon admission was associated with old age, male gender, diabetes mellitus, ischemic heat disease, hypertension and congestive heart failure. The serum albumin in patients with severe renal failure was significantly low, with a mean of 2-9 mg/dL. Urinary tract infections were more prevalent in patients with mild to severe renal failure, while intravenous line infections, bacterial endocarditis and soft and skin tissue infections were more common in patients with normal renal function. In the 279 patients with severe renal failure the mortality rate was significantly higher (50%) compared to patents with mild to moderate renal failure and patients with normal renal function (21% and 26% respectively, p = 0.0001). Multiple regression analysis revealed that pre-existing serum creatinine > 3 mg/dL was significantly associated with death attributable to bacteremia (OR = 1.7, 95% CI 1.0-2.7). In conclusion, adult bacteremic patients with pre-existing serum creatinine above 3 mg/dL upon admission are at increased risk of mortality due to bacteremia than patients with normal or mild to moderate renal failure.


Subject(s)
Bacteremia/etiology , Bacteremia/mortality , Renal Insufficiency/complications , Aged , Female , Humans , Male , Prognosis , Prospective Studies
5.
Ann Plast Surg ; 44(1): 14-7, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10651360

ABSTRACT

The high number of reconstructive dilemmas brought about by more aggressive treatment of congenital malformations and burns has created the need for large quantities of donor skin for local coverage. Tissue expansion with external ports has become part of the authors' surgical armamentarium. From January 1996 to November 1998 the authors placed 34 expanders in 28 patients to correct congenital malformations or burn sequelae. The average patient age at the time of operation was 6.3 years, the average time of expansion was 49 days, and minimal and maximal expansion volume was 60 and 600 cc respectively. Serial injection was carried out every 2 days until total expansion was achieved, and the expanded area was two to three times as wide as the recipient defect. No major complications occurred. The most serious complications of infection and erosion of tissue overlying the device occurred in 17.6% of patients. There are several advantages to this technique: less tissue dissection, painless injections, shorter operating time, and early detection of leaks. Lozano ST, Drucker MZ. Use of tissue expanders with external ports.


Subject(s)
Burns/surgery , Craniofacial Abnormalities/surgery , Tissue Expansion Devices , Tissue Expansion/methods , Adolescent , Child , Child, Preschool , Female , Humans , Male , Postoperative Complications , Treatment Outcome
6.
AJR Am J Roentgenol ; 173(6): 1651-5, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10584815

ABSTRACT

OBJECTIVE: The objective of this study was to examine the effect of breast density and age on screening mammograms with false-positive findings. MATERIALS AND METHODS: The study sample was taken from the Washington State Mammography Tumor Registry, which links data from participating radiologists with the Puget Sound Cancer Surveillance System and the Washington State Cancer Registry. Participants (n = 73,247) were women 35 years old and older who underwent screening mammography for which an assessment and a four-category density rating were coded. A total of 46,340 mammograms were sampled to avoid interpreter bias. In this study of false-positive mammograms, only women with no diagnosis of breast cancer within 12 months of the index mammogram were included. Logistic regression was used to estimate the odds ratios of a false-positive mammogram being associated with each category of breast density or age, adjusting for the other factor as a covariate. RESULTS: After controlling for breast density, we found that the risk of a false-positive mammogram was not affected by age (p = 27). However, the trend of increasing risk of a false-positive mammogram with increasing breast density was highly significant (p < .001). Women with extremely dense breast tissue were almost two times more likely to have a false-positive mammogram than were women with fatty breast tissue. This effect persisted after controlling for age. CONCLUSION: Breast density, not age, is an important factor when predicting risk of a false-positive mammogram. Breast density should be considered when educating individual women on the risks and benefits of screening mammography.


Subject(s)
Breast Neoplasms/diagnostic imaging , Mammography , Mass Screening , Adult , Age Factors , Aged , False Positive Reactions , Female , Humans , Middle Aged
7.
Cancer ; 85(11): 2410-23, 1999 Jun 01.
Article in English | MEDLINE | ID: mdl-10357412

ABSTRACT

BACKGROUND: Mammographic and physical examination assessments of the response of locally advanced breast carcinoma (LABC) to neoadjuvant therapy have been shown to be inaccurate. The authors studied the feasibility and accuracy of [technetium 99m]-sestamibi (MIBI) for monitoring the response of patients with LABC to neoadjuvant chemotherapy. METHODS: Patients receiving neoadjuvant chemotherapy for LABC underwent prone lateral scintimammography before therapy, after 2 months of therapy, and close to the completion of chemotherapy (presurgery) if chemotherapy continued for >3 months. Images were analyzed both qualitatively and quantitatively using the lesion-to-normal breast MIBI uptake ratio (L:N). Imaging results were compared with the clinical response and the pathologic response as determined from the posttherapy surgical specimen. RESULTS: A total of 32 patients (29 who were assessable for primary tumor response and 28 who were assessable for lymph node response) were included in the study. The mean change in the primary tumor L:N MIBI uptake ratio after 2 months of chemotherapy was -35% for clinical responders and +17% for nonresponders (P<0.001). Patients achieving a pathologic primary tumor macroscopic complete response (CR) had a mean change in uptake on the presurgical scan of -58% versus -18% for patients with a partial response (P<0.005). A decrease of > or =40% in the MIBI uptake ratio identified CRs with 100% sensitivity and 89% specificity. Pretherapy imaging predicted axillary lymph node metastases in 85% of patients ultimately found to have > or =1 positive lymph nodes at surgery, but was less accurate in identifying residual lymph node disease after therapy (55% sensitivity and 75% specificity). CONCLUSIONS: MIBI imaging accurately assessed the response to neoadjuvant chemotherapy in patients with LABC. Further studies are needed to determine the role of MIBI in this group of patients.


Subject(s)
Breast Neoplasms/drug therapy , Mammography/methods , Monitoring, Physiologic , Neoadjuvant Therapy , Technetium Tc 99m Sestamibi , Adult , Aged , Aged, 80 and over , Breast Neoplasms/pathology , Feasibility Studies , Female , Humans , Lymphatic Metastasis , Middle Aged , Reproducibility of Results , Treatment Outcome
8.
Clin Infect Dis ; 28(4): 822-7, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10825045

ABSTRACT

In a retrospective study, 80 episodes of nontyphoid salmonella (NTS) bacteremia in children were compared with 55 episodes in adults over a 10-year period. The study disclosed major differences in the predisposition, clinical presentation, and outcome as well as the microbiology of NTS bacteremia in relation to age. Adults were more likely than children to have predisposing diseases (95% vs. 15%, respectively; P < .0001) and to receive prior medications (95% vs. 23%, respectively; P < .0001), particularly immunosuppressive agents (58% vs. 5%, respectively; P < .0001). In most adults (67%), NTS infection presented as a primary bacteremia and was associated with a high incidence of extraintestinal organ involvement (34%) and a high mortality rate (33%). In children, NTS bacteremia was usually secondary to gastroenteritis (75%) and caused no fatalities. Although group D Salmonella (78%) and the serovar Salmonella enteritidis were the predominant isolates from adults, the emergence of infections due to group C Salmonella (46%) and the serovar Salmonella virchow in children was noted.


Subject(s)
Aging , Bacteremia/epidemiology , Salmonella Infections/epidemiology , Salmonella/isolation & purification , Adolescent , Adult , Age Distribution , Aged , Aged, 80 and over , Bacteremia/microbiology , Bacteremia/pathology , Case-Control Studies , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Prognosis , Retrospective Studies , Salmonella Infections/microbiology , Salmonella Infections/pathology
9.
J Intern Med ; 244(5): 379-86, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9845853

ABSTRACT

OBJECTIVES: To test whether empirical antibiotic treatment that matches the in vitro susceptibility of the pathogen (appropriate treatment) improves survival in patients with bloodstream infections; and to measure the improvement. DESIGN: Observational, prospective cohort study. SETTING: University hospital in Israel. SUBJECTS: All patients with bloodstream infections detected during 1988-94. INTERVENTIONS: None. MAIN OUTCOME MEASURES: In-hospital fatality rate and length of hospitalization. RESULTS: Out of 2158 patients given appropriate empirical antibiotic treatment, 436 (20%) died, compared with 432 of 1255 patients (34%) given inappropriate treatment (P = 0.0001). The median durations of hospital stay for patients who survived were 9 days for patients given appropriate treatment and 11 days for patients given inappropriate treatment. For patients who died, the median durations were 5 and 4 days, respectively (P < 0.05), for both comparisons. In a stratified analysis, fatality was higher in patients given inappropriate treatment than in those given appropriate treatment in all strata but two: patients with infections caused by streptococci other than Streptococcus gr. A and Streptoccocus pneumoniae (odds ratio (OR) of 1.0, 95% confidence interval (95% CI) 0.4-2.5); and hypothermic patients (OR = 0.9, 95% CI = 0.3-2.4). Even in patients with septic shock, inappropriate empirical treatment was associated with higher fatality rate (OR = 1.6, 95% CI = 1.0-2.7). The highest benefit associated with appropriate treatment was observed in paediatric patients (OR = 5.1, 95% CI = 2.4-10.7); intra-abdominal infections (OR = 3.8, 95% CI = 2.0-7.1); infections of the skin and soft tissues (OR = 3.1, 95% CI = 1.8-5.6); and infections caused by Klebsiella pneumoniae (OR = 3.0, 95% CI = 1.7-5.1) and S. pneumoniae (OR = 2.6, 95% C = 1.1-5.9). On a multivariable logistic regression analysis, the contribution of inappropriate empirical treatment to fatality was independent of other risk factors (multivariable adjusted OR = 1.6, 95% CI = 1.3-1.9). CONCLUSION: Appropriate empirical antibiotic treatment was associated with a significant reduction in fatality in patients with bloodstream infection.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Sepsis/drug therapy , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Logistic Models , Male , Middle Aged , Prospective Studies , Risk Factors , Sepsis/mortality , Treatment Outcome
10.
World J Surg ; 22(4): 394-7; discussion 397-8, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9523522

ABSTRACT

Local anesthetics have several effects on wound healing. In experimental studies, procaine at high concentrations has been proved to retard healing in surgical wounds by diminishing the synthesis of mucopolysaccharides and hence probably collagen. Other studies have shown that lidocaine and bupivacaine inhibit collagen synthesis in fibroblast tissue cultures in rats. This study was designed to evaluate the effect of lidocaine on wound healing. An experimental, prospective, comparative, crossover and double-blind study was designed. Forty male guinea pigs, weighing 300 to 600 g, were randomly assigned to two groups. In control group A (20 animals), skin and subcutaneous tissue in a clean wound were incised and infiltrated with regular saline solution; in group B 20 animals were infiltrated with 1% lidocaine. All animals were sacrificed on day 8 and evaluated for breaking strength, number of collagen fibers by morphometry, and histologic examination of collagenization, edema, vascularity, and presence of acute and chronic inflammatory cells. The histopathologic appearance of tissues infiltrated with lidocaine did not vary consistently in relation to collagenization, edema, or acute and chronic inflammatory processes. The mean breaking strength between both groups was not statistically significant (p = 0.120). Important statistical differences were observed in vascularity (p < 0.003) and morphometric results (p < 0.001), where collagen was found in small amounts in the lidocaine group. The results of this study suggest that local infiltration of lidocaine produces significant histopathologic changes, but it does not substantially alter wound healing as there were no differences in the breaking strength of the wounds.


Subject(s)
Anesthetics, Local/pharmacology , Lidocaine/pharmacology , Skin/drug effects , Wound Healing/drug effects , Animals , Collagen/biosynthesis , Collagen/drug effects , Cross-Over Studies , Double-Blind Method , Guinea Pigs , Male , Prospective Studies , Random Allocation , Skin/injuries , Skin/metabolism
11.
Med Pediatr Oncol ; 30(2): 95-100, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9403017

ABSTRACT

The purpose of this work was to assess the feasibility of home intravenous antibiotic treatment (HIAT) for febrile episodes in immune-compromised (neutropenic, splenectomized), low-risk pediatric patients. Thirty hematology-oncology patients who presented to our emergency room from January 1993 to January 1995 and who suffered from a febrile episode and were considered at low risk for septic complications were immediately discharged on HIAT. Patients were followed for at least 3 weeks after recovery. Patients and parents were retrospectively questioned about adverse effects and about their degree of satisfaction with home treatment. Patients who required hospitalization during this period were considered unresponsive to HIAT and were analyzed for causes and adverse effects. Thirteen out of 60 (22%) febrile episodes, or eight out of 42 (19%) episodes of fever and neutropenia eventually led to hospitalization. Pseudomonas species infections were associated with the highest rate of unresponsiveness (88%). A central venous catheter infection developed in two cases following HIAT (two cases out of 640 days of therapy). No other complications were identified. No infection-related morbidity was observed. Patients and parents were highly satisfied with HIAT and wanted to use it again, if necessary. Immediate discharge on HIAT for low-risk pediatric immune-compromised patients suffering from a febrile episode is feasible, safe, and well accepted by patients and families. Patients who are found to have Pseudomonas infections should probably be hospitalized. Our results are preliminary and must be confirmed by a prospective, randomized trial before definite recommendations can be made.


Subject(s)
Anti-Bacterial Agents/administration & dosage , Fever/drug therapy , Home Infusion Therapy , Neutropenia/drug therapy , Adolescent , Adult , Anti-Bacterial Agents/adverse effects , Child , Child, Preschool , Female , Humans , Immunocompromised Host , Infant , Infusions, Intravenous , Male , Patient Satisfaction , Treatment Outcome
12.
Eur J Clin Microbiol Infect Dis ; 16(8): 563-7, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9323466

ABSTRACT

In order to determine the epidemiology, microbiology, and outcome of bacteraemia originating in the urinary tract in hospitalised patients, a prospective study was conducted in a large general hospital in Israel. Data from all patients with bacteraemia were collected prospectively, and a subgroup of patients with bacteraemia secondary to urinary tract infection was analysed. There were 702 episodes of bacteraemia secondary to urinary tract infection during a five-year period (33.9% of all episodes of bacteraemia). The mean age of the patients was 76 years, and the male:female ratio was 0.9:1.0. The most common pathogens were Escherichia coli (52%), Klebsiella spp. (14%), and Proteus spp. (9%). Pseudomonas spp. were isolated from 8% of all patients, from 19% of those who had received antibiotics, and from 15% of males. Enterococcus spp. were isolated from 4% of males but from no females. Five percent of the episodes were polymicrobial, and 16% of the infections were hospital acquired. On logistic multivariate regression analysis, predictors of mortality were: hospitalisation in a medical department, hospital-acquired infection, inappropriate empiric antibiotic treatment, presence of decubitus ulcer(s), respiratory or renal failure, and elevated urea and decreased albumin levels.


Subject(s)
Bacteremia/epidemiology , Cross Infection/epidemiology , Urinary Tract Infections/epidemiology , Adolescent , Adult , Age Distribution , Aged , Aged, 80 and over , Analysis of Variance , Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/microbiology , Child , Child, Preschool , Cross Infection/microbiology , Female , Hospitals, University/statistics & numerical data , Humans , Incidence , Infant , Israel/epidemiology , Male , Middle Aged , Predictive Value of Tests , Prognosis , Prospective Studies , Risk Factors , Sex Distribution , Urinary Tract Infections/drug therapy , Urinary Tract Infections/microbiology
13.
Antimicrob Agents Chemother ; 41(5): 1127-33, 1997 May.
Article in English | MEDLINE | ID: mdl-9145881

ABSTRACT

The aim of the present study was to test whether the combination of a beta-lactam drug plus an aminoglycoside has advantage over monotherapy for severe gram-negative infections. Of 2,124 patients with gram-negative bacteremia surveyed prospectively, 670 were given inappropriate empirical antibiotic treatment and the mortality rate in this group was 34%, whereas the mortality rate was 18% for 1,454 patients given appropriate empirical antibiotic treatment (P = 0.0001). The mortality rates for patients given appropriate empirical antibiotic treatment were 17% for 789 patients given a single beta-lactam drug, 19% for 327 patients given combination treatment, 24% for 249 patients given a single aminoglycoside, and 29% for 89 patients given other antibiotics (P = 0.0001). When patients were stratified according to risk factors for mortality other than antibiotic treatment, combination therapy showed no advantage over treatment with a single beta-lactam drug except for neutropenic patients (odds ratio [OR] for mortality, 0.5; 95% confidence interval [95% CI], 0.2 to 1.3) and patients with Pseudomonas aeruginosa bacteremia (OR, 0.7; 95% CI, 0.3 to 1.8). On multivariable logistic regression analysis including all risk factors for mortality, combination therapy had no advantage over therapy with a single beta-lactam drug. The mortality rate for patients treated with a single appropriate aminoglycoside was higher than that for patients given a beta-lactam drug in all strata except for patients with urinary tract infections. When the results of blood cultures were known, 1,878 patients were available for follow-up. Of these, 816 patients were given a single beta-lactam drug, 442 were given combination treatment, and 193 were given a single aminoglycoside. The mortality rates were 13, 15, and 23%, respectively (P = 0.0001). Both on stratified and on multivariable logistic regression analyses, combination treatment showed a benefit over treatment with a single beta-lactam drug only for neutropenic patients (OR, 0.2; 95% CI, 0.05 to 0.7). In summary, combination treatment showed no advantage over treatment with an appropriate beta-lactam drug in nonneutropenic patients with gram-negative bacteremia.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Drug Therapy, Combination/therapeutic use , Gram-Negative Bacterial Infections/drug therapy , Adolescent , Adult , Aged , Aged, 80 and over , Aminoglycosides , Anti-Bacterial Agents/administration & dosage , Child , Child, Preschool , Female , Gram-Negative Bacterial Infections/mortality , Humans , Infant , Infant, Newborn , Logistic Models , Male , Middle Aged , Prospective Studies , Risk Factors , Treatment Outcome , beta-Lactams
14.
Scand J Infect Dis ; 29(1): 71-5, 1997.
Article in English | MEDLINE | ID: mdl-9112302

ABSTRACT

Of 4,289 episodes of bacteremia detected in 3,631 patients, septic shock was diagnosed in 453 episodes (10.5%). In 56% of shock episodes, septic shock developed more than 24 h after the first positive blood culture was taken. In a logistic regression analysis, variables predictive of septic shock were: advanced age [odds ratio (OR) of 1.015 for an increment of 1 year]; renal failure as an underlying disorder (OR = 1.47); neutropenia (OR of 2.26); curtailed functional capacity (OR of 1.54 for an increment of 1 category); unknown source of infection (OR = 1.66); anaerobic (OR = 2.86), polymicrobial bacteremia (OR = 1.54), or pathogens other than Streptococcus viridans (OR = 0.08 for Streptococcus viridans). The in-hospital mortality associated with septic shock was 80% vs 21% in episodes of bacteremia without shock, and shock episodes accounted for 31% of all deaths. The fatality rate in shock patients given appropriate empiric antibiotic treatment was 74.9% vs 84.7% in patients given inappropriate treatment (p = 0.01). Judging by the present results, host factors are more important determinants for development of septic shock in bacteremic patients than the type of pathogen. Even in patients with shock, appropriate empiric antibiotic treatment was associated with an improved chance of survival.


Subject(s)
Bacteremia/complications , Shock, Septic/etiology , Adult , Aged , Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/epidemiology , Chi-Square Distribution , Female , Hospital Mortality , Humans , Israel/epidemiology , Logistic Models , Male , Middle Aged , Prognosis , Risk Factors , Shock, Septic/drug therapy , Shock, Septic/epidemiology , Statistics, Nonparametric
15.
Eur J Biochem ; 240(3): 570-5, 1996 Sep 15.
Article in English | MEDLINE | ID: mdl-8856056

ABSTRACT

Five major polypeptides of 70, 50, 47, 19 and 17 kDa and four minor polypeptides (100, 65, 45 and 39 kDa) become phosphorylated when clathrin-coated vesicles (CCV) from zucchini hypocotyls are incubated in [gamma 32P]Mg-ATP. After dissociation with 0.5 M Tris/HCl the CCV coat polypeptides were subjected to gel filtration in order to separate clathrin triskelions from beta-adaptin-containing fractions. Only the latter bore kinase activities, with phosphorylated polypeptides of 39 kDa in addition to the 50, 19-kDa and 17-kDa polypeptides just mentioned. Heparin, an inhibitor of casein kinase II, permitted the phosphorylation of only the 19-kDa and 17-kDa polypeptides. Staurosporine, an inhibitor of protein kinase c-like activities, prevented the phosporylation of the 70-kDa polypeptide. When recombined with the triskelions the beta-adaptin fractions achieved the phosphorylation of the 45-kDa and 70-kDa polypeptides. Because of its heat stability and calcium-binding properties we interpret the 45-kDa polypeptide as being a clathrin light chain. Antibodies raised against the 70-kDa group of heat-shock proteins (Hsp70) recognize a 70-kDa polypeptide in the beta-adaptin-containing fractions. Because this polypeptide only phosphorylates in the presence of triskelions we consider it to be the uncoating ATPase, which is known to aggregate upon dissociation of the CCV coat. Our results therefore indicate that zucchini CCV contain a number of phosphorylable polypeptides equivalent to the beta, mu and sigma adaptins of bovine brain. Just as in bovine brain CCV a casein-kinase-II-like activity is associated with the zucchini CCV 50/47-kDa polypeptides, further pointing to their identity as plant mu2/mu1 adaptin equivalents.


Subject(s)
Clathrin/metabolism , Coated Vesicles/enzymology , Phosphotransferases/metabolism , Vegetables/enzymology , Animals , Brain/metabolism , Cattle , Coated Vesicles/metabolism , Kinetics , Membrane Proteins/metabolism , Molecular Weight , Phosphorylation , Phosphotransferases/chemistry , Plant Proteins/chemistry , Plant Proteins/metabolism , Species Specificity , Vegetables/metabolism
16.
Eur J Pediatr ; 155(7): 545-50, 1996 Jul.
Article in English | MEDLINE | ID: mdl-8831075

ABSTRACT

UNLABELLED: To identify bacteraemic children who are at increased risk of inappropriate empiric antibiotic therapy, we performed univariate and multivariate analyses of prospectively-studied bacteraemic episodes. Appropriateness of therapy was defined according to the in vitro susceptibility of the isolate. Inappropriate empiric therapy was found in 38% of 516 bacteraemic episodes and was associated with higher mortality. The rate of inappropriate treatment was lower in neonates and infants (28% and 33%, respectively) but higher in children 1- to 5-years old (51%, P = 0.0029). The rate was dependent on the source of bacteraemia (range, 18%-70%, P = 0.0092), underlying conditions (range, 26%-53%, P = 0.0001), the specific paediatric section in which the child was hospitalized (range, 24%-70%, P = 0.0002), and the causative micro-organism (range, 15%-75%, P < 0.0001). Four clinical variables that independently and significantly affected the rate of inappropriate antibiotic treatment were identified by multivariate stepwise logistic regression analysis (odds ratios in parentheses): hospital-acquired bacteraemia (2.3), age of 1- to 5-years (2.1), cytotoxic therapy (1.8) and presence of central i.v. line (1.6). CONCLUSION: We defined bacteraemic children who are at risk of inappropriate empiric antibiotic therapy. Special efforts are needed to improve their treatment and consequently their outcome.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Bacteremia/drug therapy , Bacteremia/epidemiology , Medication Errors , Adolescent , Age Factors , Analysis of Variance , Bacteremia/microbiology , Child , Child, Preschool , Cross Infection/drug therapy , Drug Utilization Review , Humans , Infant , Infant, Newborn , Israel/epidemiology , Logistic Models , Odds Ratio , Prospective Studies , Recurrence , Risk Factors
17.
Pediatr Infect Dis J ; 15(2): 117-22, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8822283

ABSTRACT

BACKGROUND: Hospital- and community-acquired Gram-negative bacteremia is a significant cause of mortality and morbidity in pediatric medical centers. Gram-negative organisms are isolated in > 50% of pediatric patients with bacteremia. OBJECTIVES: To analyze clinical and epidemiologic variables associated with Gram-negative bacteremia in a tertiary children's medical center. METHODS: A 6-year prospective study of children with Gram-negative bacteremia in a tertiary care children's medical center in Israel. RESULTS: Three hundred seventy-four episodes of Gram-negative bacteremia were studied during 6 years. The predominant isolates were Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli, which accounted for 109, 81 and 79 episodes (26, 20 and 19%), respectively. Of all episodes 43% occurred in neonates and infants younger than 2 years and 47% were hospital-acquired. Underlying conditions mainly acute leukemia and lymphoma, were present in 55% of the patients. Urinary tract infection followed by lower respiratory tract infection were the most common identified sources of bacteremia. Central intravenous catheters were associated with 53% of the episodes. The crude mortality was 11.4%. Increased mortality was significantly associated with acute leukemia, neutropenia, hospital-acquired infections and previous corticosteroid therapy (P = 0.03, 0.003, 0.006 and 0.01, respectively). Increased antibiotic resistance of hospital-acquired vs community-acquired isolates was noted; 44 to 77% resistance of nosocomial Klebsiella and Enterobacter sp. to second and third generation cephalosporins and 18% were resistant to amikacin. CONCLUSIONS: Klebsiella pneumoniae is currently the most common organism causing Gram-negative bacteremia in children. Because of the relatively high resistance of Gram-negative organisms to second and third generation cephalosporins, we suggest that empiric antibiotic therapy for Gram-negative bacteremia include a combination of an aminoglycoside and an anti-Pseudomonas beta-lactam.


Subject(s)
Bacteremia/epidemiology , Cross Infection/epidemiology , Adolescent , Bacteremia/diagnosis , Bacteremia/drug therapy , Child , Child, Preschool , Cross Infection/diagnosis , Female , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/epidemiology , Humans , Incidence , Infant , Male , Prospective Studies , Risk Factors
18.
Scand J Infect Dis ; 28(2): 139-42, 1996.
Article in English | MEDLINE | ID: mdl-8792479

ABSTRACT

To determine recent trends in the incidence and severity, group A streptococcal (GAS) bacteremia was studied over the last 14 years (1981-1994). There were 116 events of GAS bacteremia, representing 1.7% of all bacteremic episodes, without an increase in recent years. A total of 108 patients were available for study. Underlying conditions were found in 95 patients (88%), including mainly malignant diseases, chronic obstructive pulmonary disease, congestive heart failure and diabetes mellitus. A source of the bacteremia was noted in 71 patients (66%), with skin and soft tissue infection being the major portal of entry. All isolates were susceptible to penicillin. Overall mortality was 21%. Mortality had not increased in recent years, but depended significantly on several clinical factors: increased age; admission temperature; source of bacteremia (highest for GAS bacteremia without an identified source); and underlying conditions (highest for diabetes mellitus and chronic pulmonary disease, absent for patients with no underlying disease). This study shows that neither the incidence nor the severity of GAS bacteremia has increased in recent years. Severity is significantly affected by the source of bacteremia and the presence of underlying conditions.


Subject(s)
Bacteremia/epidemiology , Streptococcal Infections/epidemiology , Streptococcus pyogenes/isolation & purification , Adolescent , Adult , Age Distribution , Aged , Aged, 80 and over , Bacteremia/mortality , Child , Child, Preschool , Female , Hospitals, General , Humans , Incidence , Infant , Infant, Newborn , Israel/epidemiology , Male , Middle Aged , Prospective Studies , Risk Factors , Severity of Illness Index , Streptococcal Infections/mortality , Survival Rate
19.
J Antimicrob Chemother ; 36(4): 681-95, 1995 Oct.
Article in English | MEDLINE | ID: mdl-8591943

ABSTRACT

Four hundred and forty-one and 1048 episodes of bacteraemia were prospectively surveyed over a period of 18 months in two hospitals, a 450 bed community hospital and a 900 bed tertiary care urban university hospital. Incidence of bacteraemia was 2.18 per 1000 hospitalization days (10.1 per 1000 admissions) in the community hospital and 2.64 per 1000 hospitalization days (12.0 per 1000 admissions (P < 0.004)) in the university hospital. Sixty six and 62% of episodes of bacteraemia were community acquired. The majority of bacteraemic episodes originated on the internal medicine wards of both hospital--46.7% and 58.7% respectively; the incidence of bacteraemia in the medical divisions of both hospitals was 23.1 and 17.5 per 1000 admissions respectively (P < 0.01). Overall mortality rates were 22% and 26.7% respectively. 39.9% and 44% of all isolates were Gram-positive pathogens. Escherichia coli was the commonest Gram-negative pathogen in both hospitals, particularly the community hospital--47.5% vs 32.8% (P < 0.005) of all Gram-negative pathogens, while Pseudomonas spp. were significantly more common in the university hospital--18.5% vs 11.8% (P < 0.02). Non-enterococcal streptococci were more common in the community hospital while enterococci were far more common at the university hospital--15.1% vs 1% of all Gram-positive pathogens (P < 0.05). Staphylococcus epidermidis was more common among the community hospital Gram-positive bacteraemias--31.1% vs 18.6% (P < 0.005). For almost all genera and species, antibiotic resistance was higher at the university hospital. Twenty nine point four per cent of Staphylococcus aureus isolates from the university hospital were methicillin resistant compared to 2.4% at the community hospital (P < 0.005). 29.4% of all Streptococcus pneumoniae isolates at the university hospital were penicillin resistant while no resistance was found at the community hospital. A high resistance rate to ofloxacin was found at the university hospital among S. aureus and Pseudomonas sp. Sources of bacteraemia did not differ significantly between the two hospitals. In conclusion, although outcome did not differ significantly for the two hospitals, there were significant differences between blood culture isolates in these two different settings. These differences may influence clinical decision-making about antibiotic therapy for patients in these hospitals.


Subject(s)
Bacteremia/epidemiology , Hospitals, Community , Hospitals, University , Adult , Bacteremia/drug therapy , Bacteremia/microbiology , Bacteremia/mortality , Community-Acquired Infections/drug therapy , Community-Acquired Infections/epidemiology , Community-Acquired Infections/microbiology , Community-Acquired Infections/mortality , Cross Infection/drug therapy , Cross Infection/epidemiology , Cross Infection/microbiology , Cross Infection/mortality , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/epidemiology , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/mortality , Gram-Positive Bacterial Infections/drug therapy , Gram-Positive Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/microbiology , Gram-Positive Bacterial Infections/mortality , Hospitals, Rural , Hospitals, Urban , Humans , Israel/epidemiology , Prospective Studies , Treatment Outcome
20.
JAMA ; 274(10): 807-12, 1995 Sep 13.
Article in English | MEDLINE | ID: mdl-7650804

ABSTRACT

OBJECTIVE: To delineate long-term survival after an episode of bacteremia or fungemia and risk factors for mortality. DESIGN: Cohort study. SETTING: A 900-bed university hospital in Israel. PATIENTS: Study group comprising 1991 patients 18 years of age or older in whom bacteremia or fungemia were detected between March 1988 and October 1992, and a control group comprising 1991 inpatients without any infectious diseases, matched for age, sex, department, date of admission, and underlying disorders. INTERVENTIONS: None. MEASUREMENTS: Interval from the date of the first positive blood culture (study group) or from date of the identical hospital day (in the matched control patient) to the date of death as recorded in the Israeli National Population registry or, if alive, to June 1, 1994. RESULTS: The median age of patients was 72 years. In the study group, the mortality rate was 26% at 1 month, 43% at 6 months, 48% at 1 year, and 63% at 4 years, and the median survival was 16.2 months. In the control group, the mortality rate was 7% at 1 month, 27% at 1 year, and 42% at 4 years, and the median survival was greater than 75 months (P < .001). Factors significantly and independently associated with mortality in bacteremic patients were functional class (median survival, 0.5 month in bedridden patients), septic shock (median survival, 0.2 month), serum albumin (median survival, 1.1 months in the lowest quartile), serum creatinine (median survival, 2.9 months in the highest quartile), age (median survival, 2.9 months in the highest quartile [age > 80 years]), inappropriate empirical antibiotic treatment (median survival, 4.9 months), nosocomial infection (median survival, 9.6 months), and malignancy (median survival, 2.4 months). CONCLUSIONS: Bacteremia is associated with high short-term mortality, but also a sign of severely curtailed long-term prognosis, especially in patients with low functional capacity, low serum albumin, high serum creatinine, nosocomial infections, malignancy, inappropriate antimicrobial treatment, and septic shock and in elderly patients.


Subject(s)
Bacteremia/mortality , Fungemia/mortality , Adult , Age Distribution , Aged , Cohort Studies , Female , Humans , Israel , Male , Matched-Pair Analysis , Middle Aged , Multivariate Analysis , Survival Analysis , Survivors
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