Impact of adding Saccharomyces strains on fermentation, aerobic stability, nutritive value, and select lactobacilli populations in corn silage.

Bacterial inoculants can improve the conservation and nutritional quality of silages. Inclusion of the yeast Saccharomyces in the diet of dairy cattle has also been reported to be beneficial. The present study assessed the ability of silage to be used as a means of delivering Saccharomyces strains to ruminants. Two strains of Saccharomyces cerevisiae (strain 1 and 3)and 1 strain of Saccharomyces paradoxus (strain 2) were inoculated (10(3) cfu/g) individually onto corn forage that was ensiled in mini silos for 90 d. Fermentation characteristics, aerobic stability, and nutritive value of silages were determined and real-time quantitative PCR (RT-qPCR) was used to quantify S. cerevisiae, S.paradoxus, total Saccharomyces, fungal, and bacterial populations. Fermentation characteristics of silage inoculated with S1 were similar to control silage. Although strain 3 inoculation increased ash and decreased OM contents of silage (P = 0.017), no differences were observed in nutrient composition or fermentation profiles after 90 d of ensiling. Inoculation with Saccharomyces had no detrimental effect on the aerobic stability of silage. In vitro DM disappearance, gas production, and microbial protein synthesis were not affected by yeast inoculation.Saccharomyces strain 1 was quantified throughout ensiling, whereas strain 2 was detected only immediately after inoculation. Saccharomyces cerevisiae strain 3 was quantified until d 7 and detectable 90 d after ensiling. All inoculants were detected and quantified during aerobic exposure. Inoculation with Saccharomyces did not alter lactobacilli populations. Saccharomycetales were detected by RT-qPCR throughout ensiling in all silages. Both S. cerevisiae and S. paradoxus populations increased during aerobic exposure, demonstrating that the density of these yeast strains would increase between the time that silage was removed from storage and the time it was fed.

Mechanics and spiral formation in the rat cornea.

During the maturation of some mammals such as mice and rats, corneal epithelial cells tend to develop into patterns such as spirals over time. A better understanding of these patterns can help to understand how the organ develops and may give insight into some of the diseases affecting corneal development. In this paper, a framework for explaining the development of the epithelial cells forming spiral patterns due to the effect of tensile and shear strains is proposed. Using chimeric animals, made by combining embryonic cells from genetically distinguishable strains, we can observe the development of patterns in the cornea. Aggregates of cell progeny from one strain or the other called patches form as organs and tissue develop. The boundaries of these patches are fitted with logarithmic spirals on confocal images of adult rat corneas. To compare with observed patterns, we develop a three-dimensional large strain finite element model for the rat cornea under intraocular pressure to examine the strain distribution on the cornea surface. The model includes the effects of oriented and dispersed fibrils families throughout the cornea and a nearly incompressible matrix. Tracing the directions of critical strain vectors on the cornea surface leads to spiral-like curves that are compared to the observed logarithmic spirals. Good agreement between the observed and numerical curves supports the proposed assumption that shear and tensile strains facilitate sliding of epithelial cells to develop spiral patterns.

Reduced cellular susceptibility to in vitro HIV infection is associated with CD4+ T cell quiescence.

BACKGROUND: HIV preferentially establishes productive infection in activated CD4+ T cells. Since proportions of activated CD4+ T cells vary between individuals, this study aimed to determine if individuals with a greater proportion of activated CD4+ T cells would be more susceptible to in vitro HIV infection. METHODOLOGY/PRINCIPAL FINDINGS: Unstimulated peripheral blood mononuclear cells (PBMC) from various donors were inoculated with HIV(ML1956)in vitro. HIV replication was evaluated by HIV p24 ELISA of culture supernatants and intracellular staining for HIV p24, which was detected by flow cytometry. Baseline T cell phenotypes and infected cell phenotypes were also evaluated by flow cytometry. Ex vivo phenotyping at the time of blood draw showed that elevated T cell activation and reduced Tregs were associated with increased cellular susceptibility to in vitro infection. Furthermore, the infected CD4+ T cell population was enriched for activated cells. CONCLUSION/SIGNIFICANCE: These data suggest that CD4+ T cell quiescence provides an environment less conducive to the establishment of HIV infection by limiting the pool of activated target cells.

Cervical HIV-specific IgA in a population of commercial sex workers correlates with repeated exposure but not resistance to HIV.

We conducted a comprehensive cross-sectional analysis of total and HIV-specific cervical antibody levels in HIV-1-resistant, uninfected, and infected women in order to examine the role of HIV-specific antibody responses in the female genital tract and examine the effect on antibody levels of various epidemiologic factors in this population. Cervical lavages were collected from 272 subjects of the Pumwani commercial sex worker cohort. Total and HIV-specific genital tract IgA and IgG levels were measured using an ELISA and correlated with behavioral and demographic factors. No significant difference was seen between cervical HIV-specific IgA levels in infected, uninfected, and resistant individuals, nor were any correlations between cervical HIV-specific IgA and neutralization capacity or viral shedding seen. We did, however, note increased HIV-specific IgA in HIV-negative women with four or more clients per day, and decreased HIV-specific IgA in both long-term nonprogressors and long-term survivors. These results show that there is not a strong cohort-wide correlation between HIV-specific cervical IgA levels and resistance to infection by HIV-1 as previously believed, but there is a correlation between exposure to HIV and HIV-specific cervical IgA. Our findings do not preclude the possibility that functional differences in the cervical IgA of HEPS women may play a role in resistance, but argue that HIV-specific responses may not be a universal protective factor. They also indicate that resistance to HIV is a complex condition related to more factors than exposure. Further studies of correlates of immune protection in these individuals would be beneficial to the field.

BMI-referenced cut points for pedometer-determined steps per day in adults.

BACKGROUND: The goal of this study was to establish preliminary criterion-referenced cut points for adult pedometer-determined physical activity (PA) related to weight status defined by body mass index (BMI). METHODS: Researchers contributed directly measured BMI and pedometer data that had been collected (1) using a Yamax-manufactured pedometer, (2) for a minimum of 3 days, (3) on ostensibly healthy adults. The contrasting groups method was used to identify age- and gender-specific cut points for steps/d related to BMI cut points for normal weight and overweight/obesity (defined as BMI <25 and >or=25 kg/m2, respectively). RESULTS: Data included 3127 individuals age 18 to 94 years (976 men, age = 46.8 +/- 15.4 years, BMI = 27.3 +/- 4.9; 2151 women, age = 47.4 +/- 14.9 years, BMI = 27.6 +/- 6.4; all gender differences NS). Best estimated cut points for normal versus overweight/obesity ranged from 11,000 to 12,000 steps/d for men and 8000 to 12,000 steps/d for women (consistently higher for younger age groups). CONCLUSIONS: These steps/d cut points can be used to identify individuals at risk, or the proportion of adults achieving or falling short of set cut points can be reported and compared between populations. Cut points can also be used to set intervention goals, and they can be referred to when evaluating program impact, as well as environmental and policy changes.

Development of a novel allo-independent HIV-1 virus preparation for use in immunoassays.

The study of the immunologic response to whole human immunodeficiency virus type 1 (HIV-1) antigen is limited by the presence of highly immunogenic human leukocyte antigen (HLA) alloantigens on the envelope of wild type virus. This paper outlines the production of HIV-1 infectious virions free of HLA for use as whole viral antigens in immunoassays. An infectious molecular clone of HIV-1 was transfected into the K-562 cell line, which does not express HLA on the cell surface. After a 30-day selection period, to ensure stable transfection, cells and culture supernatants were analyzed for productive HIV-1 infection and virion infectivity. An enzyme-linked immunosorbent assay (ELISA) confirmed the presence of p24 in the culture supernatants. Molecular confirmation of HIV-1 transfection was achieved by gene amplification. Flow cytometric analysis was used to identify gp120 on the surface of the infected cells. Viral supernatants were tested for HIV infectivity in peripheral blood mononuclear cells (PBMCs). The usefulness of this viral preparation as whole virus antigens was validated using PBMCs from HIV-infected individuals. These results indicate the successful production of HIV-1 infectious virions, which do not have HLA molecules on their viral envelope, and demonstrate their utility for immunoassays.

HIV-1-specific cellular immune responses among HIV-1-resistant sex workers.

The goal of the present study was to determine whether there were HIV-1 specific cellular immune responses among a subgroup of women within a cohort of Nairobi prostitutes (n = 1800) who, despite their intense sexual exposure to HIV-1, are epidemiologically resistant to HIV-1 infection. Of the 80 women defined to be resistant, 24 were recruited for immunological evaluation. The HIV-1-specific T-helper responses were determined by IL-2 production following stimulation with HIV-1 envelope peptides and soluble gp120. Cytotoxic T lymphocyte responses were determined by lysis of autologous EBV-transformed B cell lines infected with control vaccinia virus or recombinant vaccinia viruses containing the HIV-1 structural genes env, gag and pol. Resistant women had significantly increased HIV-1 specific T-helper responses, as determined by in vitro IL-2 production to HIV-1 envelope peptides and soluble glycoprotein 120, compared with low-risk seronegative and HIV-1-infected controls (P < or = 0.01, Student's t-test). Seven of the 17 (41%) resistant women showed IL-2 stimulation indices > or = 2.0. HIV-1-specific CTL responses were detected among 15/22 (68.2%) resistant women compared with 0/12 low-risk controls (Chi-squared test, P < 0.001). In the two resistant individuals tested, the CTL activity was mediated by CD8+ effectors. Many HIV-1-resistant women show evidence of HIV-1-specific T-helper and cytotoxic responses. These data support the suggestion that HIV-1-specific T-cell responses contribute to protection against HIV-1 infection.

HIV type 1 resistance in Kenyan sex workers is not associated with altered cellular susceptibility to HIV type 1 infection or enhanced beta-chemokine production.

A small group of women (n = 80) within the Nairobi-based Pumwani Sex Workers Cohort demonstrates epidemiologic resistance to HIV-1 infection. Chemokine receptor polymorphisms and beta-chemokine overproduction have been among the mechanisms suggested to be responsible for resistance to HIV-1 infection. This study attempts to determine if any of those mechanisms are protecting the HIV-1-resistant women. Genetic analysis of CCR5 and CCR3 from the resistant women demonstrated no polymorphisms associated with resistance. Expression levels of CCR5 among the resistant women were shown to be equivalent to that found in low-risk seronegative (negative) controls, while CXCR4 expression was greater among some of the resistant women. In vitro infection experiments showed that phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs) from resistant women were as susceptible to infection to T cell- and macrophage-tropic North American and Kenyan HIV-1 isolates as were the PBMCs from negative controls. No significant difference in circulating plasma levels of MIP-1alpha and MIP-1beta were found between the resistant women and negative or HIV-1-infected controls. In vitro cultures of media and PHA-stimulated PBMCs indicated that the resistant women produced significantly less MIP-1alpha and MIP-1beta than did negative controls and no significant difference in RANTES levels were observed. In contrast to studies in Caucasian cohorts, these data indicate that CCR5 polymorphisms, altered CCR5 and CXCR4 expression levels, cellular resistance to in vitro HIV-1 infection, and increased levels of beta-chemokine production do not account for the resistance to HIV-1 infection observed among the women of the Pumwani Sex Workers Cohort.

A single mutation in the M-subunit of Rhodospirillum rubrum confers herbicide resistance.

Cells of the photosynthetic bacterium Rhodospirillum rubrum were rendered resistant against the inhibitor 2-(1-phenyl)ethylamino-3-propionylamino-4-cyano-thiazole (PPCTH). Electron transport in reaction centers prepared from one of the mutants (M6) was neither inhibited by PPCTH and other NH-thiazoles nor terbutryn. These inhibitors are known to bind at the Q(B) site of the L-subunit. Compared to the wild type, chromatophores from M6 exhibited strongly altered Q(B)- Fe2+ and Q(A)- Fe2+ EPR signals. Inhibitor resistance is due to a mutation in the bacterial reaction center M-subunit, where Glu234 is exchanged against Lys. This is the first example of an inhibitor resistance in the Q(B) site caused by a mutation in the M-subunit.

End-stage renal disease: a proving ground for quality improvement in health care.

This article chronicles the health care quality improvement efforts that relate to patients with end stage renal disease (ESRD). The emphasis is on quality improvement as a management system as opposed to the quality improvements that resulted from strictly technical dialysis-related issues. The government has exercised considerable oversight on the ESRD program because of its growth and cost. History has shown that quality assurance (QA) has had little effect on improving quality or decreasing cost. The philosophy, methods, and tools of continuous quality improvement (CQI) have been shown to work in health care. CQI is a management system that offers hope for higher quality affordable health care. Computer technology is at last sophisticated enough to permit the collection of large amounts of clinical data at the point of care. This will permit CQI methods and tools to be applied generally at reasonable costs. Physicians in general and nephrologists in particular are beginning to understand the managed care environment. They are beginning to understand the paradigm shift that is required to effect the changes necessary for physicians to assume their leadership role in health care. This article reviews the quality efforts of the past and present. It discusses the strengths and weaknesses of efforts to improve quality and lastly presents a vision for the future.

High-dose versus standard-dose epinephrine treatment of cardiac arrest after failure of standard therapy.

STUDY OBJECTIVE: To assess the efficacy of high-dose epinephrine (HDE) compared with standard-dose epinephrine (SDE) in emergency department patients in cardiac arrest after SDE failed to improve asystole or ventricular fibrillation. DESIGN: Prospective, multicenter, blinded, controlled trial. SETTING: Eight academic center emergency departments. PATIENTS: One hundred forty patients treated for cardiac arrest. MEASUREMENTS AND MAIN RESULTS: Primary outcomes were either improvement in cardiac rhythm or return of spontaneous circulation (ROSC). Of the 140 patients enrolled, 78 received HDE and 62 received SDE. Of the 34 patients with ventricular fibrillation, 3 were resuscitated with HDE and 2 with SDE (p = 0.60). Of those with asystole, ROSC occurred in 12 of HDE and 5 of SDE recipients (p = 0.11). No patient had return of significant neurologic function and none survived to hospital discharge. Overall, there was no advantage to HDE after failure of SDE. CONCLUSION: Our results are similar to those of controlled clinical trials comparing HDE with SDE in cardiac arrest.

Improving clinical processes: one dialysis facility's experiences.

Northeast Louisiana Dialysis Center implemented continuous quality improvement (CQI) to improve the quality of care delivered to end stage renal disease (ESRD) patients treated by hemodialysis in their facility. The unit chose to address normalization of calcium and phosphorus and parathyroid hormone (PTH), anemia, nutrition, adequacy of dialysis and dialyzer reuse as well as performance benchmarks by the Health Care Financing Administration (HCFA) core indicators. This article presents the results obtained and the methodology used in this improvement effort. The article also presents nine principles the authors believe necessary for a successful CQI program.

Normal human cord blood B cells can produce high affinity IgG antibodies to dsDNA that are recognized by cord blood-derived anti-idiotypic antibodies.

It is possible to identify, in Epstein-Barr virus-transformed normal human cord blood B cell populations, cells present at a low frequency that produce IgG antibodies specific for dsDNA. By cloning out these B cells as immortalized monoclonal cell lines, it could be shown that the antibodies were the products of CD5 positive B cells. Two monoclonal anti-dsDNA antibodies were derived from cell lines T52 and A7 and these were further characterized as anionic (pI approximately 6.4) IgG4 kappa antibodies that bound with affinities of 7.18 x 10(9) l/mol and 3.28 x 10(9) l/mol, respectively, to dsDNA but did not bind to ssDNA. These affinities were similar to those of polyclonal IgG anti-dsDNA antibodies from lupus patients, which ranged from 1 x 10(9) -8.9 x 10(10) l/mol. Both T52 and A7 monoclonal anti-dsDNA antibodies were recognized by cord blood-derived IgM antibodies. These IgM antibodies were not rheumatoid factors but bound to the F(ab')2 of A7 and T52 while failing to recognize T50, which is an autologous IgG4 kappa monoclonal antibody without specificity for dsDNA. A cloned B cell line A24 generated from the same cord blood sample as A7 produced an IgM monoclonal antibody that bound to the heavy chains of T52 and A7, but not T50 on Western blot and inhibited the binding of these antibodies to dsDNA. A7 and T52 competitively inhibited each other in their binding to the anti-idiotype A24, and A24 inhibited the binding to dsDNA of some polyclonal IgG anti-dsDNA antibodies purified from sera of lupus patients. The level of inhibition of binding of these antibodies to dsDNA was directly proportional to the levels of expression of the idiotype recognized by A24 on these antibodies. The normal human cord blood, therefore, may contain cells that form an idiotype/anti-idiotype network in which the idiotype is expressed on IgG antibodies with specificity for dsDNA and the anti-idiotype is an IgM antibody that binds to a heavy chain idiotope in such a way as to interfere with its interaction with dsDNA. The presence of a similar idiotype on some polyclonal anti-dsDNA antibodies in lupus that are similarly inhibitable by the cord blood-derived anti-idiotype raises the possibility that this network may persist in later life and perhaps become dysfunctional in systemic lupus erythematosus.

The post-concussional state: neurophysiological aspects.

BACKGROUND: Twenty-six young men admitted to an Accident and Emergency Department for observation following a minor closed head injury (post-traumatic amnesia (PTA) less than 12 hours) were investigated within 24 hours of admission (day 0) and followed up at 10 days, 6 weeks and 1 year after the trauma. METHOD: Investigations at day 0 included physical examination, completion of post-concussional symptom and stress-arousal checklists, computerised EEG (CEEG) and auditory brainstem evoked potential (BAEP) recordings. These were repeated at ten days and six weeks. At 12 months follow-up, the Present State Examination (PSE) was carried out and a further post-concussional symptom checklist completed. RESULTS: Post-concussional symptomatology declined progressively from day 0 but half had residual symptoms at 1 year. Seventy-two per cent ran an acute course with recovery by 6 weeks, 8% chronic unremitting course and 20% initially improved but had an exacerbation of symptoms between 6 weeks and 12 months. The CEEG alpha-theta ratios decreased significantly between days 0 and 10, reaching a baseline thereafter. Measures of CEEG recovery from all channels correlated with symptom counts at six weeks; the slower the recovery the greater the symptoms. A relative delay in left temporal recovery was associated with residual psychiatric morbidity (PSE ID scores) at 12 months. Prolonged central brainstem conduction times occurred in 27% of patients at day 0. These correlated positively with PTA and degree of psychiatric morbidity (PSE ID scores) at 12 months. CONCLUSIONS: Symptom chronicity was accompanied by continuing brainstem dysfunction, while the degree of transient cortical dysfunction appeared to have a direct influence in the intensity of early organic symptom reaction to the trauma. Levels of perceived stress at the time of the injury, or afterwards, were not related to symptom formation.

Validation of criterion-referenced standards for tests of arm and shoulder girdle strength and endurance.

The purpose of this study was to establish criterion-referenced standards for selected tests of arm and shoulder girdle strength and endurance for college females. Several popular tests of arm and shoulder girdle strength and endurance were administered to equal numbers of trained and untrained college females (N = 92) to generate data for the analysis. The contrasting groups method (Berk, 1976; Safrit & Wood, 1990) yielded the following criterion cutoff scores that classified college females as trained or untrained on the basis of regular upper arm and shoulder girdle resistance training: pull-ups = .5 repetitions (reps), 90 degrees push-ups = 16 reps, flexed arm hang = 5 s, seated chest press = 24.5 kg, seated biceps curl = 16.4 kg, seated lat pull (latissimus dorsi pull-down) = 38.2 kg, absolute strength index = 86.4 kg, and relative strength index = .6 kg per kg of body weight. Cross-validation of the standards on an independent sample of college females (N = 112) suggested stability of the cutoff scores for pull-ups, flexed arm hang, and relative strength.

Fenoldopam mesylate versus sodium nitroprusside in the acute management of severe systemic hypertension.

Thirty-three patients with severe systemic hypertension defined as a diastolic blood pressure (DBP) > or = 120 mm Hg were randomized in a single-blind fashion to be treated with either intravenous fenoldopam mesylate (FNP) or sodium nitroprusside (NTP). Fenoldopam mesylate and NTP infusion rates began at 0.1 microgram/kg/minute and 0.5 microgram/kg/minute, respectively and were titrated to achieve a goal DBP of between 95 and 110 mm Hg; or a reduction of at least 40 mm Hg if the baseline DBP was > 150 mm Hg. Fenoldopam mesylate (n = 15) reduced blood pressure from 217/145 +/- 6/5 to 187/112 +/- 6/3 mm Hg (P < .001) at an average infusion rate of 0.5 +/- 0.1 microgram/kg/minute. The average time to achieve goal DBP with FNP was 1.5 +/- 1.4 hours. Nitroprusside (n = 18) reduced blood pressure from 210/136 +/- 5/2 to 172/103 +/- 6/2 mm Hg (P < .001) at an average infusion rate of 1.2 +/- .24 micrograms/kg/minute. Nitroprusside response time averaged 2 +/- 2.5 hours. There was no significant difference between the magnitude of effect seen with either FNP or NTP; nor was there any difference observed in the adverse effect rates of the two agents. Fenoldopam mesylate and NTP demonstrate similar overall efficacy in the treatment of severe systemic hypertension.

The postconcussional syndrome: social antecedents and psychological sequelae.

The study consisted of a prospective investigation of 45 consecutively admitted patients who had sustained a mild head injury. In all cases the duration of post-traumatic amnesia was less than 24 hours. Head injury patients had an average of three adverse life events in the year preceding injury compared with 1.5 for controls. Using the PSE, 39% of the group were diagnosed psychiatric cases at six weeks after the injury. For cases the mean level of chronic social difficulties (3.3) was four times that for non-cases (0.8). Six months after injury, 28% of the head injury group had three or more symptoms. These chronic cases were on average ten years older than those whose symptoms had remitted. Chronic cases had, on average, three social difficulties, twice as many as found among those whose symptoms had remitted. The emergence and persistence of the postconcussional syndrome are associated with social adversity before the accident. While young men are most at risk of minor head injury, older women are most at risk of chronic sequelae.

Spontaneous pneumomediastinum: clinical and natural history.

STUDY OBJECTIVE: To evaluate the clinical characteristics and natural history of patients presenting with spontaneous pneumomediastinum. DESIGN: A retrospective case series was conducted to identify patients diagnosed with spontaneous pneumomediastinum. ICD-9 discharge codes were used for 1984 to 1990 at two institutions, and emergency department records of a third hospital were reviewed for 1981 to 1986. Clinical features, interventions, complications, setting, etiology, symptoms, and length of hospital stay were recorded. SETTING: Three university tertiary care hospitals. PARTICIPANTS: All ED patients more than 12 years old with a diagnosis of spontaneous pneumomediastinum. INTERVENTIONS: None. RESULTS: Seventeen cases were identified. Age range was 15 to 41 years (mean, 25 years). Presenting symptoms were chest pain in eight (47%), dyspnea in three (18%), both symptoms in three (18%), and neither in three (18%). Three patients complained only of throat discomfort. Nine (52%) had a Hamman's crunch, 11 (65%) had subcutaneous emphysema, and two (11%) had a small pneumothorax. Five (29%) were smokers, and five (29%) had normal esophograms. Thirteen of 17 (76%) cases were associated with illicit inhalation drug use. Twelve cases (70%) had history of a "Valsalva-type" maneuver. All but three were admitted to a hospital, with a mean stay of 2.5 days (range, one to six). No patient suffered complications or required interventions for spontaneous pneumomediastinum. Specifically, no patient developed a subsequent pneumothorax or airway compromise. The three patients not admitted were followed up by telephone contact. All did well with rapid resolution of their symptoms. CONCLUSION: Most spontaneous pneumomediastinum cases occur in the setting of inhalational drug use. One hundred percent of patients will have a symptom directly related to the spontaneous pneumomediastinum, with 82% presenting with either dyspnea or chest pain. Most (88%) will present with either subcutaneous emphysema or a Hamman's crunch on examination. Simple spontaneous pneumomediastinum has a very benign course and does not require hospitalization. Serial radiographs, likewise, did not change the medical management of spontaneous pneumomediastinum.