Prospective study in critically ill non-neutropenic patients: diagnostic potential of (1,3)-β-D-glucan assay and circulating galactomannan for the diagnosis of invasive fungal disease

Diagnosis of invasive fungal disease (IFD) in patients under intensive care is challenging. Circulating biomarkers, (1,3)-β-D-glucan (BG) and galactomannan (GM), were prospectively assessed in 98 critically ill patients at risk of IFD. There were 11 cases of invasive aspergillosis (IA; 4 proven and 7 probable), 9 cases of proven invasive candidiasis (IC), 1 case of mixed proven IC and probable IA, 1 case of proven zygomycosis, and 1 case of mixed mycelial proven IFD. In all IA cases there was no significant difference when the area under the receiver operating characteristic curve (AUC) of GM (0.873 [95%CI, 0.75–0.99]) and BG (0.856 [95% CI, 0.71–0.99]) were compared (p = 0.871). The AUC for BG in IC and for the rest of the IFD cases was 0.605 (95% CI, 0.39–0.82) and 0.768 (95% CI, 0.63–0.90) respectively. Positive BG (40%) predated blood culture (n = 3) and abdominal pus (n = 1) a mean of 3.25 days before Candida was grown. In patients with IFD caused by molds, BG appeared a mean of 5.65 days before culture results. For the diagnosis of patients at risk of IC, BG has shown a high NPV (94.5%), with positive results also predating blood cultures in 30% of patients. In conclusion, early BG results permit a timely initiation of antifungal therapy in patients at risk of IFD.     

Rules for identifying patients at increased risk for candidal infections in the surgical intensive care unit: approach to developing practical criteria for systematic use in antifungal prophylaxis trials.

The high rates of invasive candidiasis among intensive care unit (ICU) patients suggest that antifungal prophylaxis might be of value, but rules identifying patients who would best benefit are not established. Based on a retrospective study of 327 patients who stayed in a surgical ICU for > or = 4 days and had an 11.0% rate of invasive candidiasis, we sought to identify useful predictive rules. As prior work suggests that prompt initiation of prophylaxis is of value, we required our rules to be based on patient data routinely available during the week prior to ICU admission through the third day of the ICU stay. Patients with any combination of diabetes mellitus, new onset hemodialysis, use of total parenteral nutrition, or receipt of broad-spectrum antibiotics had an invasive candidiasis rate of 16.6% versus a 5.1% rate for patients lacking these characteristics (P = 0.001). Fifty-two percent of patients staying > or = 4 days in the ICU met this rule and the rule captured 78% of the patients who eventually developed invasive candidiasis. Risk-stratified antifungal prophylaxis in the ICU is possible. Validation of these results in other types of ICU is now needed.     

Diagnosis of invasive candidiasis by a dot immunobinding assay for Candida antigen detection.

A dot immunobinding assay which uses a polyclonal rabbit anti-Candida immunoglobulin G as the primary antibody and colloidal gold coated with goat anti-rabbit immunoglobulin G as the secondary antibody for the detection of Candida cytoplasmic antigens is described. It was able to detect as little as 1 ng of total Candida protein per ml when a cytoplasmic extract of Candida albicans was seeded into buffer and 10 ng/ml when the same extract was seeded into pooled human serum. Serial serum samples from four groups of patients were assayed for Candida antigen: (i) 22 patients with candidemia, (ii) 16 patients at high risk for invasive candidiasis, (iii) 3 patients with other deep mycoses, and (iv) 50 hospitalized patients at low risk for serious Candida infection. Of the 22 candidemic patients, 19 had invasive candidiasis and 3 had transient candidemia. Antigenemia was detected in 16 of the 19 patients with invasive candidiasis (including patients with C. albicans, Candida tropicalis, Candida glabrata, Candida krusei, and Candida parapsilosis) and in 4 of 16 patients at high risk for invasive candidiasis. There was no detectable antigen in 12 high-risk control patients, 3 patients with transient candidemia, 3 patients with other deep mycoses, and 50 relatively low-risk patients. The sensitivity for detecting invasive disease in candidemic patients and specificity for all patients studied were 84.2 and 94.4%, respectively. The positive predictive value was 80%; the negative predictive value was 95.7%. The sensitivity for neutropenic patients with invasive disease was 85.7%. This assay is rapid and accurate and appears to be useful in identifying candidemic patients with invasive candidiasis.     

Factors related to survival and treatment success in invasive candidiasis or candidemia: a pooled analysis of two large, prospective, micafungin trials

Crude and attributable mortality rates in patients with candidemia and invasive candidiasis remain unacceptably high. It is important to reach a more complete understanding of the risk factors underlying poor outcomes in patients with invasive Candida infections. Micafungin therapy has been assessed in two phase 3 trials compared to either liposomal amphotericin B or caspofungin. The availability of this large dataset allows the analyses of non-drug factors associated with survival and treatment success. A multivariate regression analysis was performed on data from the two trials separately and as a pooled analysis (N = 1,070). Analysis outcomes were survival at 42 days post-initiation of therapy and treatment success. For the pooled analysis, treatment success was significantly more likely for candidemia than invasive candidiasis. Both survival and treatment success were significantly less likely for the non-removal of catheter versus removal, Asian-Indians versus Caucasians, APACHE II score >20 to ≤30 and >30 versus ≤20, age ≥70 years versus <50 years, baseline corticosteroids, and persistent neutropenia. Survival was also significantly less likely for treatment in other regions versus North America and for patients with renal failure at baseline. These findings help to define non-antifungal drug factors that may impact survival and treatment success in invasive candidiasis or candidemia.     

Role of polymorphic variants of gene of inducible NO synthase <Emphasis Type="Italic">NOS2</Emphasis> in brain infarction in patients with acute ischemic stroke

Cerebrovascular diseases, including stroke, are an important problem in public health. Stroke development depends on external factors and the individual genetic specificity of the patient. Excessive NO production by inducible NO synthase (iNOS) damages brain tissue at various stages of the disease. The goal of this work was to study the role of four polymorphic variants of gene of inducible NO synthase iNOS (−2447C/G, −1659C/T, −0.7(TTTA)n I/D, S608L (150C/T)) in brain infarction in patients with acute ischemic stroke. A statistically significant correlation between S608L (150C/T) polymorphism and infarction dynamics was observed during days 1–3 and 7–21 after infarction. These parameters correlate to the neurological status, which is estimated using the Orgogozo scale during days 1–7 of disease development. It was demonstrated that the C150C genotype was associated with ischemic focus propagation, regardless of its volume and neurological status by Orgogozo scale in patients with acute stroke. In the case of low initial volume, it was observed that the C150C genotype had an effect on ischemic damage during days 1–3.     

Creatine supplementation improves muscular performance in older women

Muscle power and strength decrease with age leading to reduced independence and increased health risk from falls. Creatine supplementation can increase muscle power and strength. The purpose of this study was to examine the effects of 7 days of creatine supplementation on body composition, muscular strength, and lower-body motor functional performance in older women. Thirty 58–71 year old women performed three test sessions (T1–T3) each separated by one week. Each session consisted of one repetition maximum tests for bench press and leg press, and isometric hand-grip, tandem gait, upper-body ergometer, and lower-body ergometer tests. Following T2, subjects were assigned to a creatine monohydrate (0.3 g kg body mass⁻¹ for 7 days) (CR: 63.31 ± 1.22 year, 160.00 ± 1.58 cm, 67.11 ± 4.38 kg) or a placebo (PL: 62.98 ± 1.11 year, 162.25 ± 2.09 cm, 67.84 ± 3.90 kg) supplementation group. CR significantly (P < 0.05) increased bench press (1.7 ± 0.4 kg), leg press (5.2 ± 1.8 kg), body mass (0.49 ± 0.04 kg) and fat free mass (0.52 ± 0.05) and decreased completion time on the functional tandem gait tests from T2–T3. No significant changes were found for PL on any of the measured variables. No adverse side-effects were reported by either group. Short-term creatine supplementation resulted in an increase in strength, power, and lower-body motor functional performance in older women without any adverse side effects     

Improved accuracy and extended flow range for a Fleisch pneumotachograph

A large linear flow range and a small instrumental dead space volume are incompatible properties for a pneumotachometer (PTM). The linearity of a Fleisch number 2 PTM is studied for flows up to 6 litre s⁻¹ (nominal range 0–2 litre s⁻¹) with various up- and downstream geometries. It is hypothesised that using an array of calibration factors (conductance; flow/pressure), instead of a single calibration factor over the entire flow range, could improve accuracy and also extend the applicable flow range. The conductance against pressure characteristics are calculated with a previously described weighted averaging technique based on multiple strokes from a precision syringe. A single conductance value gives stroke volume errors in the range of −5 to 3% (0–2 litre s⁻¹) and −6 to 11% (0–6 litre s⁻¹) for validation using the same geometry as for calibration. The pressure dependent conductance improves accuracy to within −3% and 1% independent of flow range. However, for validation using a different geometry than for calibration, errors range from −5% to +8%. The degree of non-linearity varies between the geometries (range 3–15%) and is highest when using a one-directional valve upstream of the PTM and a Y-shaped connector. In conclusion, a pressure-dependent conductance improves accuracy and can also be used to extend the applicable flow range up to at least three times the nominal flow range.     

Histopathological subclassification of triple negative breast cancer using prognostic scoring system: five variables as candidates

We attempted to subclassify triple negative breast cancer (TNBC) cases into subgroups according to clinical outcome or prognosis of TNBC patients using archival specimens. We analyzed 102 Japanese cases of invasive TNBC who underwent surgery between January 1998 and December 2007. The clinicopathological factors and clinical information were retrospectively retrieved from reviewing the charts of the patients. Immunohistochemical staining was performed for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), epidermal growth factor receptor 1 (EGFR1), CK5/6, CK14, Ki-67, and CD31 for microvessel density (MVD). Median follow-up time of the patients was 68.5 months. Multivariable analysis demonstrated that pathologic node status was the most significantly associated with relapse-free survival (RFS) and breast cancer-specific survival (BCSS) of these patients. Pathological tumor size, basal-like type, Ki-67 labeling index (LI) and MVD were also independently associated with RFS and BCSS. Based on these results, we devised the risk score system reflecting hazard ratios of these prognostic factors above. With this system, TNBC patients in this study were classified into three subgroups (low-risk group: score 0–3, intermediate-risk group: score 4–7 and high-risk group: score 8–10). The significant difference of RFS and BCSS was detected among these three different subgroups of the patients (p < 0.05). We propose the risk score system, which incorporated pathologic nodal status, size of the primary tumor, the presence or absence of basal-like features, Ki-67 LI, and MVD in order to predict postoperative clinical course of the Japanese TNBC patients.     

Effects of menstrual phase on performance and recovery in intense intermittent activity

Game sport and training require repeated high intensity bursts. This study examined differences between high intensity, intermittent work in two phases of the menstrual cycle. Six physically active young women (age 19–29) performed 10 6-s sprints on a cycle ergometer in both the mid-follicular (FP) (days 6–10) and late-luteal phases (LP) (days 20–24) of the menstrual cycle. Work, power, oxygen intake parameters, and capillarized blood lactate were measured. Data are analyzed using the Friedman and Wilcoxon matched pairs tests. There was no difference between menstrual phases in peak 6-s power (6.8(0.6) W kg⁻¹ in FP, 6.9(0.6) W kg⁻¹ in LP), the drop off in work (1.2(3.5) J kg⁻¹ in FP and 1.0(2.7) J kg⁻¹ in LP), or in the sprint (23.7 (1.5) mL kg⁻¹ min⁻¹ in LP and 24.3(2.4) mL kg⁻¹ min⁻¹ in FP). Capillarized blood lactate was also similar in both phases of the menstrual cycle both at 1 min (9.2 (2.7) mmol L⁻¹ in FP, 9.2 (3.1) mmol L⁻¹) and at 3 min (9.0 (2.2) mmol L⁻¹ in FP, 9.2 (2.2) mmol L⁻¹ in LP). However, the average 6-s work was greater in the LP (39.3 (3.4) J kg⁻¹) than during the FP (38.3 (3.1) J kg⁻¹) (P=0.023). The recovery was also greater in the LP than the FP (26.3 (2.4) mL kg⁻¹ min⁻¹ in LP, 25.0 (2.6) mL kg⁻¹ min⁻¹ in FP, P=0.023). Average work over a series of sprints and the consumed between sprints may be slightly greater during the LP than the FP of the menstrual cycle.     

Granulocyte chemiluminescence response to serum opsonized zymosan particlesex vivo during long-term strenuous exercise,energy and sleep deprivation in humans

The chemiluminescence response of granulocytes to serum opsonized zymosan particles (SOZ) ex vivo was investigated during two ranger training courses lasting 7 days with continuous moderate physical activities corresponding to about 32% of maximal oxygen uptake or 35 000 kJ · 24 h⁻¹, with energy deficiency (energy supply 0-4000 kJ · 24 h⁻¹), and less than 3-h sleep during the 7 days. Significant granulocytosis in combination with a lymphopenia in peripheral blood was observed during the whole course. A priming of the granulocytes for accentuated chemiluminescence response to SOZ was observed during the first days of the course with a maximal increase on day 3 in course A (+35% of control response) and on day 1 in course B (+ 12%). Thereafter, reduced responses to SOZ compared to control values (−28% and −21% in course A and B) were observed. In course A, a group (n = 8) receiving 5000 kJ · 24 h⁻¹ of additional energy, showed a more pronounced priming (maximum +57% versus +21 % of control response) during the first days. In course B, all the cadets had 3 h of organised rest/sleep on day 5, and a second priming of the chemiluminescence response was observed on the subsequent 2 days. These data indicated that moderate, continuous, predominantly aerobic physical activities for 1–3 days around the clock primed the production of reactive oxygen species in granulocytes. This priming may be beneficial for, for example, host defence against micro-organisms, but may also contribute to inflammatory damage to normal tissues such as muscle, tendons and joints during exercise. However, when the moderate exercise continued for several more days, a down-modulation of the granulocyte response was observed. The findings of this study further support the possibility that moderate physical activity stimulates immunity, while more extreme duration of the same activities may result in a down-modulation of nonspecific (and specific) immunity.     

The effects of training intensity on muscle buffer capacity in females

We examined changes in muscle buffer capacity (βm_{in vitro}), and the lactate threshold (LT) after 5 weeks of high-intensity interval training (INT) above the LT or moderate-intensity continuous training (CON) just below the LT. Prior to and immediately after training, 16 female subjects performed a graded exercise test to determine and the LT, followed 2 days later by a resting muscle biopsy from the vastus lateralis muscle to determine βm_{in vitro}. Following baseline testing, the subjects were randomly placed into the INT (n=8) or CON training group (n=8). Subjects then performed 5 weeks of cycle training (3 days per week), performing either high-intensity INT (6–10×2 min at 120–140% LT with 1 min rest) or moderate-intensity CON (80–95% LT) training. Total training volume was matched between the two groups. After the training period, both groups had significant improvements in (12–14%; P<0.05) and the LT (7–10%; P<0.05), with no significant differences between groups. The INT group, however, had significantly greater improvements in βm_{in vitro} (25%; 123±5–153±7 μmol H⁺·g muscle dm⁻¹·pH⁻¹; P<0.05) than the CON group (2%; 130±12–133±7 μmol H⁺·g muscle dm⁻¹·pH⁻¹, P>0.05). Our results show that when matched for training volume, high-intensity interval training above the LT results in similar improvements in and the LT, but greater improvements in βm_{in vitro} than moderate-intensity continuous training below the LT. This suggests that training intensity is an important determinant of changes to βm_{in vitro}.     

Use of X-ray Tomography to Map Crystalline and Amorphous Phases in Frozen Biomaterials

The outcome of both cryopreservation and cryosurgical freezing applications is influenced by the concentration and type of the cryoprotective agent (CPA) or the cryodestructive agent (i.e., the chemical adjuvants referred to here as CDA) added prior to freezing. It also depends on the amount and type of crystalline, amorphous and/or eutectic phases formed during freezing which can differentially affect viability. This work describes the use of X-ray computer tomography (CT) for non-invasive, indirect determination of the phase, solute concentration and temperature within biomaterials (CPA, CDA loaded solutions and tissues) by X-ray attenuation before and after freezing. Specifically, this work focuses on establishing the feasibility of CT (100–420 kV acceleration voltage) to accurately measure the concentration of glycerol or salt as model CPA and CDAs in unfrozen solutions and tissues at 20°C, or the phase in frozen solutions and tissue systems at −78.5 and −196°C. The solutions are composed of water with physiological concentrations of NaCl (0.88% wt/wt) and DMEM (Dulbecco’s Modified Eagle’s Medium) with added glycerol (0–8 M). The tissue system is chosen as 3 mm thick porcine liver slices as well as 2 cm diameter cores which were either imaged fresh (3–4 h cold ischemia) or after loading with DMEM based glycerol solutions (0–8 M) for times ranging from hours to 7 days at 4°C. The X-ray attenuation is reported in Hounsfield units (HU), a clinical measurement which normalizes X-ray attenuation values by the difference between those of water and air. NaCl solutions from 0 to 23.3% wt/wt (i.e. water to eutectic concentration) were found to linearly correspond to HU in a range from 0 to 155. At −196°C the variation was from −80 to 95 HU while at −78.5°C all readings were roughly 10 HU lower. At 20°C NaCl and DMEM solutions with 0–8 M glycerol loading show a linear variation from 0 to 145 HU. After freezing to −78.5°C the variation of the NaCl and DMEM solutions is more than twice as large between −90 and +190 HU and was distinctly non-linear above 6 M. After freezing to −196°C the variation of the NaCl and DMEM solutions increased even further to −80 to +225 HU and was distinctly non-linear above 4 M, which after modeling the phase change and crystallization process is shown to correlate with an amorphous phase. In all tissue systems the HU readings were similar to solutions but higher by roughly 30 HU, as well as showing some deviations at 0 M after storage, probably due to tissue swelling. The standard deviations in all measurements were roughly 5 HU or below in all samples. In addition, two practical examples for CT use were demonstrated including: (1) glycerol loading and freezing of tissue cores and, (2) a mock cryosurgical procedure. In the loading experiment CT was able to measure the permeation of the glycerol into the sample at 20°C, as well as the evolution of distinct amorphous vs. crystalline phases after freezing to −196°C. In the mock cryosurgery example, the iceball edge was clearly visualized, and attempts to determine the temperature within the iceball are discussed. An added benefit of this work is that the density of these frozen samples, an essential property in measurement and modeling of thermal processes, was obtained in comparison to ice.     

Detection of antibodies to Candida albicans germ tubes for diagnosis and therapeutic monitoring of invasive candidiasis in patients with hematologic malignancies.

We prospectively investigated the ability of detection of antibodies to Candida albicans germ tubes (CAGT) to diagnose invasive candidiasis in 95 consecutive admissions of 73 patients with hematologic disorders undergoing intensive chemotherapy. The episodes were divided into three groups according to clinical and microbiological diagnosis. Group 1 comprised eight admissions of eight patients with invasive candidiasis. Group 2 comprised 42 admissions of 34 patients without evidence of invasive candidiasis. Group 3 comprised the remaining 45 admissions of 37 patients with febrile episodes which were not diagnosed by microbiological culture. Antibodies to CAGT were detected in 87.5% of group 1 patients. Detection of antibodies to CAGT in patients with Candida fungemia was delayed somewhat relative to the time the blood culture was positive, but antibodies to CAGT were detected earlier than a diagnosis was made in patients with deep-tissue candidiasis. Sera from 2 admissions in group 2 and 12 admissions in group 3 revealed antibodies to CAGT. At a titer of > or = 1:20, detection of antibodies to CAGT had a sensitivity of 87.5%, specificity of 95.2%, positive predictive value of 77.8%, and negative predictive value of 97.6%. Antibodies to CAGT were usually detected before beginning of empiric antifungal therapy. Titers of antibodies to CAGT were maintained in most patients who died but declined and eventually disappeared in the patients who survived. Since antibodies to CAGT were detected in all patients with tissue-proven invasive candidiasis but negative by blood culture, detection of antibodies to CAGT complemented blood cultures for diagnosis and therapeutic monitoring of patients with hematologic malignancies and invasive candidiasis.     

Fast and slowly inactivating components of Ca-channel current and their sensitivities to nicardipine in isolated smooth muscle cells from rat vas deferens

(1) Fast and slowly inactivating components of Ca-channel current were compared to clarify whether more than one type of Ca-channel exists in smooth muscle cells from rat vas deferens using the whole cell variant of the patch clamp technique. The pipette was filled with 150 mM Cs solution to eliminate outward current and Ba was used as the charge carrier for Ca-channel current. (2) When activated by a 5 s test pulse to 0 mV from a holding potential of −60 mV, the inactivation process of Ba-current was well fitted by the sum of two exponentials. The time constant of the faster inactivating component was 100–300 ms and that of the slower inactivating component was 1.5–3 s. Steadystate inactivation curves of the fast- and slow-components were very similar. (3) The inward current activated at 0 mV from −80 mV was inactivated faster than that from −30 mV. The voltage-dependencies of the peak current from holding potentials of −30 mV and −80 mV were similar. Both had voltage threshold at −30 mV and were maximal at +10 mV. (4) Low concentrations of nicardipine (10⁻⁹ to 10⁻⁷ M) preferentially inhibited the slow component while higher concentration (10⁻⁶ to 10⁻⁵ M) were required to block the fast component. The current activated from a holding potential of −30 mV was almost fully suppressed by 10⁻⁷ M nicardipine whereas that from −80 mV was blocked only slightly. The voltage dependencies of the peak currents before and during the superfusion with nicardipine (10⁻⁷ M) were similar although the peak amplitude was suppressed in the presence of the drug. (5) These results suggest that the existence of either (a) two populations of Ca channels that differ in the time course of inactivation and the sensitivity to nicardipine, but have nearly identical dependence on membrane potential or (b) one population of Ca channel having two different states of inactivation and the sensitivity of nicardipine, in rat vas deferens.     

Seven days’ around the clock exhaustive physical exertion combined with energy depletion and sleep deprivation primes circulating leukocytes

Both exhaustive physical exertion and starvation have been reported to induce depression of immune function. The aim of the present study was to investigate the inflammatory environment and state of activation and mediator-producing potential of circulating leukocytes during prolonged physical activity with concomitant energy and sleep deprivation. Eight well-trained males were studied during 7 days of semi-continuous physical activity. Sleep was restricted to about 1 h/24 h, energy intake to 1.5– 3.0 MJ/24 h. Blood was drawn at 07.00 a.m. on days 0, 2, 4, and 7. Plasma levels of inflammation markers were measured. The response of circulating leukocytes to lipopolysaccharide (LPS; 1 μg mL⁻¹), and the effect of added hydrocortisone (10 and 100 nmol L⁻¹), were measured in the supernatant after 3 h of incubation in an ex vivo whole blood model. Activation of leukocytes steadily increased as measured by plasma matrix metalloproteinase-9, tumour necrosis factor-α, interleukin-1β, and interleukin-6. Inhibitors of systemic inflammation were either unaltered (tissue inhibitor of matrix metalloproteinase-1) or elevated (plasma interleukin-1 receptor antagonist). Cortisol levels increased on days 2 and 4, but thereafter reverted to baseline values. The leukocytes responded to LPS activation with increasing release of inflammatory cytokines throughout the study period. The anti-inflammatory potency of hydrocortisone decreased. Prolonged multifactorial stress thus activated circulating immune cells and primed them for an increased response to a subsequent microbial challenge.     

Diurnal normobaric moderate hypoxia raises serum erythropoietin concentration but does not stimulate accelerated erythrocyte production

This study was performed to examine the effect of diurnal normobaric hypoxia on hematological parameters. Eleven healthy male volunteers were randomly selected to be in either the hypoxic group (n=6) or the control group (n=5). The hypoxic group was exposed to 8 h of normobaric hypoxia in hypoxic tent systems that elicited a target peripheral O₂ saturation of 81±2% on three consecutive days. The control group spent three consecutive 8-h days in modified tent systems that delivered normoxic air into the tent. Venous blood samples were collected before the exposure (days –5, 0), after each day of the exposure (days 1, 2, 3), and for 3 weeks after the exposure (days 7, 10, 13, 17, 24). Serum erythropoietin concentration significantly increased from 9.1±3.3 U·L⁻¹ to 30.7±8.6 U·L⁻¹ in the hypoxic group. Although there were significant increases in hematocrit (4%), hemoglobin concentration (5%), red blood cell count (4%) on day 7 in the hypoxic group, these observations were likely due to dehydration or biological variation over time. There was no significant change in early erythropoietic markers (reticulocyte counts or serum ferritin concentration), which provided inconclusive evidence of accelerated erythroid differentiation and proliferation. The results suggest that the degree of hypoxia was sufficient to stimulate increased erythropoietin production and release. However, the duration of hypoxic exposure was insufficient to propagate the erythropoietic cascade.     

Reduced bacterial dissemination and liver injury in CD14-deficient mice following a chronic abscess-forming peritonitis induced by Bacteroides fragilis

The CD14 myelomonocytic differentiation antigen plays a major role in acute Gram-negative infections with Escherichia coli; however, its role in chronic infections has not yet been analyzed. To address this question, we studied the role of CD14 in a chronic abscess-forming peritonitis, induced by Bacteroides fragilis. B. fragilis (3×10⁸ CFU/ml) were resuspended in a liquid nutrient agar and injected into the peritoneal cavity of CD14-deficient (CD14–/–) and normal C57BL/6J (CD14+/+) mice, respectively. After 3 days there was a severe phlegmonous intra-abdominal inflammation in both groups. After 7 days an abscess-forming peritonitis developed and by 14 days the infectious foci were compartimentalized. These obser-vations were indistinguishable between CD14–/– and CD14+/+ mice. Although no differences were seen in abscess formation, CD14–/– mice were able to clear B. fra-gilis more efficiently from the blood than CD14+/+ mice. After 3, 7, and 14 days blood cultures were B. fragilis positive in 11% (1/9), 20% (2/10), and 0% (0/9) in CD14–/– compared with 90% (9/10), 78% (7/9), and 20% (2/10) in CD14+/+ mice, respectively (P<0.05). Furthermore, although the infection resulted in hepatocellular necrosis and severe hepatitis in both groups, at day 14 the liver cell damage was more severe in CD14+/+ than in CD14–/– mice (P<0.05). These results show that the chronic abscess formation induced by B. fragilis capsular polysaccharides is CD14 independent; however, bacterial clearance and/or dissemination and liver cell damage are at least partially influenced by CD14-dependent mechanisms. Received: 15 September 1998     

Dissociation of force production from MHC and actin contents in muscles injured by eccentric contractions

The primary purpose of this study was to determine the relationship between myosin heavy chain (MHC) and actin contents and maximum isometric tetanic force (Po) in mouse extensor digitorum longus (EDL) muscles following eccentric contraction-induced injury. Po and protein contents were measured in injured (n=80) and contralateral control (n = 80) EDL muscles at the following time points after in vivo injury: sham, 0, 0.25, 1, 3, 5, 14, and 28 days. P_o was reduced by 37 ± 2.3% to 49 ± 3.8% (p ≤ 0.05), while MHC and actin contents were unaltered from 0 to 3 days after injury. Whereas Po partially recovered between 3 and 5 days (from −49 ± 3.8% to −35 ± 3.6%), MHC and actin contents in the injured muscles declined by 19 ± 4.9% and 20 ± 5.3%, respectively, by 5 days compared with control muscles. Decrements in Po were similar to the reductions in MHC and actin contents at 14 (∼24%) and 28 (∼11%) days. Evaluation of myofibrillar and soluble protein fractions indicated significant reductions in the content of major proteins at 5 and 14 days. Immunoblots of heat shock protein 72 revealed elevations starting at 0.25 days, peaking during 1–3 days, and declining after 5days. These findings indicate that decreased contractile protein content is not related to the initial decrease in P_o. However, decreased MHC and actin contents could account for 58% of the P_o reduction at 5 days, and for nearly all the decrements in P_o from 14 to 28 days. This revised version was published online in August 2006 with corrections to the Cover Date.     

Invasive <Emphasis Type="Italic">Streptococcus pneumoniae</Emphasis> infections in children and older adults in the north of Spain before and after the introduction of the heptavalent pneumococcal conjugate vaccine

In the last two decades, an increasing trend in the incidence of pneumococcal disease in Europe has been reported. We investigated the effect of the use of the heptavalent pneumococcal conjugate vaccine (PCV7) in an area of northern Spain, where all recorded cases of invasive pneumococcal diseases (IPD) were included (n = 450; 91 between 1996–2007 in children aged <5 years and 359 between 1998–2007 in adults aged >64 years). All isolates were serotyped. In children, the overall IPD incidence did not significantly decrease after the introduction, in late 2001, of PCV7. However, the incidence of PCV7 serotypes significantly decreased by 137.2% from 31.59 cases/100,000 population in 1996–2001 to 13.42 in 2002–2007 (95% confidence interval [CI] −27.2 to −342.4%), as did the overall rates of penicillin resistance (from 45.6 to 18.6%) and multiresistance (from 30.3 to 11%). In older adults, the overall IPD incidence showed a non-significant increase due to non-PCV7 serotypes, which seemed to continue a previous trend in our region.     

Methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> (MRSA) in hospitalized children: correlation of molecular analysis with clinical presentation and antibiotic susceptibility testing (ABST) results

The molecular analysis of methicillin-resistant Staphylococcus aureus (MRSA) from 98 children admitted to the Children’s Hospital of Michigan, Detroit, MI, with serious MRSA infections during 2006–2007 was correlated with risk factors, clinical features, and antibiotic susceptibility testing (ABST) results. Isolates were characterized by staphylococcal cassette chromosome (SCC) mec type, the presence of Panton-Valentine leukocidin (PVL) genes, repetitive sequence (rep) polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE), requirement for surgical intervention, antibiograms, and response to therapy. rep-PCR was more rapid than PFGE typing and correlated well. SCCmec type IV-containing isolates caused 92.8% of all infections, but the demographics and diseases associated with subtypes IVa and IVd differed. Subtype IVa (all PFGE type USA300 and PVL-positive) was identified in 81/93 (87.1%) of patients with community-onset (CO) MRSA, including 21/35 of those with risk factors for health care-associated (HA) infection. All other clones were PVL-negative. Subtype IVd (10 isolates; 9 USA800 and 1 eMRSA15) caused mainly HA-MRSA and no skin and soft tissue infections (SSTI). Seven classic HA-MRSA strains (SCCmec types II [6; 3 USA100 and 3 USA600] and III [1; USA200]) caused HA and hospital-onset (HO) infections. Surgical intervention was required in 68/81 patients infected with USA300 and 8/17 of the others. Most USA300 were susceptible (S) to clindamycin (CD) and patients were treated with CD alone or in combination. The other isolates were generally treated with vancomycin (VA) alone or in combination. This work was presented in part at the Infectious Diseases Society of America Annual Meeting, San Diego, CA, USA, 4–7 October 2007.