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Background Cognitive dysfunction after surgery is a major issue in older adults. Here, we determined the effect of APOE4 on perioperative neurocognitive function in older patients. Methods We enrolled 140 English-speaking patients ≥60 yr old scheduled for noncardiac surgery under general anaesthesia in an observational cohort study, of whom 52 underwent neuroimaging. We measured cognition; Aβ, tau, p-tau levels in CSF; and resting-state intrinsic functional connectivity in six Alzheimer's disease-risk regions before and 6 weeks after surgery. Results There were no significant APOE4-related differences in cognition or CSF biomarkers, except APOE4 carriers had lower CSF Aβ levels than non-carriers (preoperative median CSF Aβ [median absolute deviation], APOE4 305 pg ml−1 [65] vs 378 pg ml−1 [38], respectively; P=0.001). Controlling for age, APOE4 carriers had significantly greater preoperative functional connectivity than non-carriers between several brain regions implicated in Alzheimer's disease, including between the left posterior cingulate cortex and left angular gyrus (β [95% confidence interval, CI], 0.218 [0.137–0.230]; PFWE=0.016). APOE4 carriers, but not non-carriers, experienced significant connectivity decreases from before to 6 weeks after surgery between several brain regions including between the left posterior cingulate cortex and left angular gyrus (β [95% CI], –0.196 [–0.256 to –0.136]; PFWE=0.001). Most preoperative and postoperative functional connectivity differences did not change after controlling for preoperative CSF Aβ levels. Conclusions Postoperative change trajectories for cognition and CSF Aβ, tau or p-tau levels did not differ between community dwelling older APOE4 carriers and non-carriers. APOE4 carriers showed greater preoperative functional connectivity and greater postoperative decreases in functional connectivity in key Alzheimer's disease-risk regions, which occur via Aβ-independent mechanisms.

Pharmacological ascorbate (i.e., intravenous infusions of vitamin C reaching ~ 20 mM in plasma) is under active investigation as an adjuvant to standard of care anti-cancer treatments due to its dual redox roles as an antioxidant in normal tissues and as a prooxidant in malignant tissues. Immune checkpoint inhibitors (ICIs) are highly promising therapies for many cancer patients but face several challenges including low response rates, primary or acquired resistance, and toxicity. Ascorbate modulates both innate and adaptive immune functions and plays a key role in maintaining the balance between pro and anti-inflammatory states. Furthermore, the success of pharmacological ascorbate as a radiosensitizer and a chemosensitizer in pre-clinical studies and early phase clinical trials suggests that it may also enhance the efficacy and expand the benefits of ICIs.

Platinum-based chemotherapy with or without immunotherapy is the mainstay of treatment for advanced stage non-small cell lung cancer (NSCLC) lacking a molecular driver alteration. Pre-clinical studies have reported that pharmacological ascorbate (P-AscH-) enhances NSCLC response to platinum-based therapy. We conducted a phase II clinical trial combining P-AscH- with carboplatin-paclitaxel chemotherapy.Chemotherapy naïve advanced stage NSCLC patients received 75 g ascorbate twice per week intravenously with carboplatin and paclitaxel every three weeks for four cycles. The primary endpoint was to improve tumor response per Response Evaluation Criteria in Solid Tumors (RECIST) v1.1 compared to the historical control of 20%. The trial was conducted as an optimal Simon's two-stage design. Blood samples were collected for exploratory analyses.The study enrolled 38 patients and met its primary endpoint with an objective response rate of 34.2% (p = 0.03). All were confirmed partial responses (cPR). The disease control rate was 84.2% (stable disease + cPR). Median progression-free and overall survival were 5.7 months and 12.8 months, respectively. Treatment-related adverse events (TRAE) included one grade 5 (neutropenic fever) and five grade 4 events (cytopenias). Cytokine and chemokine data suggest that the combination elicits an immune response. Immunophenotyping of peripheral blood mononuclear cells demonstrated an increase in effector CD8 T-cells in patients with a progression-free survival (PFS) ≥ 6 months.The addition of P-AscH- to platinum-based chemotherapy improved tumor response in advanced stage NSCLC. P-AscH- appears to alter the host immune response and needs further investigation as a potential adjuvant to immunotherapy.

Individuals with autism spectrum disorder (ASD) may exhibit chronic autonomic nervous system (ANS) hyperarousal (e.g., lower respiratory sinus arrhythmia and higher heart rate) compared to their typically developing peers, reflecting a chronic biological threat response. The sustained nature of this cardiac threat suggests tonic nervous system perception of threat in safe contexts. Herein, the cardiac literature in adult and child populations with ASD is reviewed and placed within a continuum of functioning in order to inform the relationship between cardiac response and functioning in ASD. Findings from this review support the relationship between chronic autonomic hyperarousal and emotional and behavioral difficulties observed in individuals with ASD.

Summary Obesity prevalence rises fastest during young adulthood when weight, diet, and physical activity may be influenced by life events, including becoming a parent, but the impact is uncertain. We searched six electronic databases to July 2019 for longitudinal studies (both sexes) aged 15 to 35 years with a prospective pre‐pregnancy/parenthood and post‐delivery outcome. Of 11 studies (across 15 papers), six studies (women only) were eligible for meta‐analysis of the difference in change in body mass index (BMI; kg/m2) between remaining without children and becoming a parent. Mean (±SD) BMI gain for non‐mothers was 2.8 ± 1.3 kg/m2 (~7.5 kg for 164‐cm woman) over 5.6 ± 3.1 years; 12.3% of baseline BMI (22.8 ± 2.5 kg/m2). Becoming a mother was associated with an additional BMI increase of 0.47 ± 0.26 kg/m2 (~1.3 kg), 4.3% of baseline BMI (22.8 ± 5.6 kg/m2); the one study including men reported no difference in change. Physical activity results were equivocal; 2/4 studies (women) and 2/2 (men) showed a greater decline in parents versus non‐parents; diet (three studies) varied by dietary measure, mostly indicating no difference. Becoming a mother is associated with 17% greater absolute BMI gain than remaining childless. Motherhood BMI gain is additional to an alarming BMI increase among young women, highlighting the need for obesity prevention among all young women, including mothers.

BACKGROUND: The Lung Volume Reduction Coil Treatment in Patients With Emphysema (RENEW) trial reported improvements in quality of life, pulmonary function, and exercise performance following endobronchial coil treatment.OBJECTIVES: The purpose of this post hoc analysis was to identify baseline predictors, including quantitative CT measures, that identify patients most likely to significantly benefit from endobronchial coil therapy.METHODS: Quantitative CT analysis by an independent radiology laboratory and a qualitative evaluation by five blinded experts of the baseline thoracic CT imaging were performed. Univariate and multivariate logistic regression analyses were performed to elucidate characteristics associated with clinical response.RESULTS: In total, 125 patients underwent coil treatment and had evaluable 12-month follow-up results. Of these, 78 patients received treatment of lobes with the highest emphysematous destruction determined by quantitative CT analysis (quantitative visual match [QVM]+), and 47 received treatment in at least one lobe that was not the most destroyed (QVM-). From the 78 patients with QVM+ treatment, a subgroup of 50 patients (64%) was identified with baseline residual volume > 200% predicted, emphysema score > 20% low attenuation area, and absence of airway disease. In this subgroup, greater lobar residual volume reduction in the treated lobes was achieved, which was associated with significant mean +/- SE improvement in FEV1 (15.2 +/- 3.1%), St. George's Respiratory Questionnaire (-12 +/- 2 points), and residual volume (-0.57 +/- 0.13 L).DISCUSSION: This post hoc analysis found that both significant hyperinflation (residual volume >= 200% predicted) and CT analysis are critical for patient selection and treatment planning for endobronchial coil therapy. Quantitative CT analysis is important to identify optimal lobar treatment and to exclude patients with insufficient emphysema (

Background As there has been a demonstrated link between sleep disturbance and suicide, the present study investigated the relationship between self-reported sleep disturbance during inpatient psychiatric treatment and clinical outcomes at discharge and six months post-discharge. Methods Participants were 2,970 adults receiving care in a long-term inpatient psychiatric hospital. Greater than 90% of inpatients reported at least mild sleep disturbance throughout treatment. Group-based trajectory modeling was used to determine patterns of sleep disturbance changes over time. Results Participants fit into distinct categories based on their trajectories of sleep problems across treatment: No Sleep Problems (10.3% of participants, no sleep problems endorsed over the course of inpatient psychiatric treatment), Resolvers (sleep problems completely resolved over the course of inpatient psychiatric treatment; 10.6%), Non-Responders (did not respond to inpatient psychiatric treatment; 35.7%) and Responders (sleep problems decreased but did not fully resolve over the course of inpatient psychiatric treatment; 43.4%). Individuals with comorbid major depressive disorder and generalized anxiety disorder were significantly more likely to demonstrate higher rates of sleep disturbance throughout inpatient psychiatric treatment and their sleep problems did not respond to treatment as usual (Non-Responders). Further, patients in the Non-Responder group had significantly more suicidal ideation and worse clinical outcomes (higher anxiety, more disability, and lower well-being) at discharge and six months post-discharge, as well as were on more medication including hypnotics throughout treatment. Conclusions Findings indicate the urgent need to design and implement inpatient psychiatry sleep protocols to not only improve sleep and clinical outcomes, but also reduce the risk for suicide post-discharge.

Respiratory syncytial virus (RSV) and the common commensal and opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) both serve as a frequent cause of respiratory infection in children. Although it is well established that some respiratory viruses can increase host susceptibility to secondary bacterial infections, few studies have examined how commensal bacteria could influence a secondary viral response. Here, we examined the impact of NTHi exposure on a subsequent RSV infection of human bronchial epithelial cells (16HBE14o-). Co-culture of 16HBE14o- cells with NTHi 2019 resulted in inhibition of viral gene expression following RSV infection. 16HBE14o- cells co-cultured with heat-killed NTHi 2019 failed to protect against an RSV infection, indicating that protection requires live bacteria. However, NTHi did not inhibit influenza A virus replication, indicating that NTHi-mediated protection was RSV-specific. Our data demonstrates that prior exposure to a commensal bacterium such as NTHi can elicit protection against a subsequent RSV infection.

Memory CD8 T cells can provide protection from re-infection by respiratory viruses such as influenza and SARS. However, the relative contribution of memory CD8 T cells in providing protection against respiratory syncytial virus (RSV) infection is currently unclear. To address this knowledge gap, we utilized a prime-boost immunization approach to induce robust memory CD8 T cell responses in the absence of RSV-specific CD4 T cells and antibodies. Unexpectedly, RSV infection of mice with pre-existing CD8 T cell memory led to exacerbated weight loss, pulmonary disease, and lethal immunopathology. The exacerbated disease in immunized mice was not epitope-dependent and occurred despite a significant reduction in RSV viral titers. In addition, the lethal immunopathology was unique to the context of an RSV infection as mice were protected from a normally lethal challenge with a recombinant influenza virus expressing an RSV epitope. Memory CD8 T cells rapidly produced IFN-γ following RSV infection resulting in elevated protein levels in the lung and periphery. Neutralization of IFN-γ in the respiratory tract reduced morbidity and prevented mortality. These results demonstrate that in contrast to other respiratory viruses, RSV-specific memory CD8 T cells can induce lethal immunopathology despite mediating enhanced viral clearance., Author summary Memory CD8 T cells have been shown to provide protection against many respiratory viruses. However, the ability of memory CD8 T cells to provide protection against RSV has not been extensively examined. Unexpectedly, mice with pre-existing CD8 T cell memory, in the absence of memory CD4 T cells and antibodies, exhibited exacerbated morbidity and mortality following RSV infection. We demonstrate that the immunopathology is the result of early and excessive production of IFN-γ by memory CD8 T cells in the lung. Our research provides important new insight into the mechanisms of how memory T cells induce immunopathology. In addition, our findings serve as an important cautionary tale against the use of epitope-based T cell vaccines against RSV.

Background Inhibitors of mTOR, such as sirolimus, have been shown to induce thymus involution and inflammatory lung disease in mice. The latter effect supports the role of this serine/threonine kinase in ameliorating lung inflammation. Other studies have shown sirolimus reduces/delays lung disease associated with various strains of influenza A virus (IAV). Thus, the effects of mTOR inhibitors on influenza infection deserve further studies. Methods Here, we examined the changes in lung viral copies, pathology and pulmonary function associated with IAV (A/PR/8/34) infection in mice treated with sirolimus. Results Body weight loss peaked between days 6–11 post-infection and was more severe in IAV-infected mice that were administered sirolimus as compared to mice that received IAV alone (p = 0.030). Natural log viral gene copies, mean ± SD per mg lung tissue, in IAV-infected mice that were administered sirolimus were 17.31 ± 1.27 on day 4, 19.31 ± 7.46 on day 10, and 0 on day 25. The corresponding number of copies in mice that received IAV alone were 18.56 ± 0.95 on day 4 (p = 0.132), 1.52 ± 1.39 on day 10 (p = 0.008), and 0 on day 25. Lung pathology was evident on days 4, 10, and 25 post infection, with mean ± SD inflammatory score of 9.0 ± 4.5 in IAV-infected mice that were administered sirolimus, as compared to 11.5 ± 4.5 (p = 0.335) in mice received IAV alone (maximum score, 26.0). Impaired lung function was evident in IAV-infected mice on days 4 and 10, as demonstrated by increased airway resistance and decreased compliance. Conclusions In this model, the effects of sirolimus on influenza infection included severe weight loss and modified viral replication, respiratory function and lung inflammation. The adverse events associated with sirolimus treatment are consistent with its potent immunosuppressive activity and, thus, preclude its use in IAV infection.

Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis and pneumonia in children under 1 y of age in the USA. The host immune response is believed to contribute to RSV-induced disease. We hypothesize that severe RSV infection in infants is mediated by insufficient regulation of the host immune response of regulatory T cells (Tregs) resulting in immunopathology. Blood and nasal aspirates from 23 RSV-infected and 17 control infants under 1 y of age were collected. Treg frequencies were determined by flow cytometry from peripheral blood mononuclear cells. Analysis of 24 cytokines was measured by multiplex assay on nasal aspirates. We demonstrate that the frequency of activated Tregs is significantly reduced in the peripheral blood of RSV-infected infants compared with age-matched controls. Surprisingly, T helper (Th)17 related cytokines including interleukin (IL)-1β, IL-17A, and IL-23 were associated with a reduction in clinical symptoms of respiratory distress. In addition, the amount of IL-33 protein in nasal washes, a cytokine important in maintaining Treg homeostasis in mucosal tissues, was decreased in RSV-infected children. These results suggest that decreased Treg numbers and an inability to properly control the host inflammatory response results in severe RSV infection.

The treatment of a child with a relapsed state acute leukemia after allogeneic stem cell transplantation (allo-SCT) is a challenge. The authors report about a child with an acute myelogenous leukemia (AML), which relapsed after allo-SCT despite immunological intervention. It was further treated with a second line chemotherapy followed by an infusion of stem cells and donor lymphocytes. Because of an immense risk for a further relapse, an immunological maintenance therapy was also performed, consisting of repetitive infusions of low doses of donor lymphocytes combined with low-dose chemotherapy. Presently, the child is in continuous complete remission and has a good quality of life.

Due to their ability to rapidly proliferate and produce effector cytokines, memory CD8+ T cells increase protection following reexposure to a pathogen. However, low inflammatory immunizations do not provide memory CD8+ T cells with a proliferation advantage over naive CD8+ T cells, suggesting that cell-extrinsic factors enhance memory CD8+ T cell proliferation in vivo. Herein, we demonstrate that inflammatory signals are critical for the rapid proliferation of memory CD8+ T cells following infection. Using murine models of viral infection and antigen exposure, we found that type I IFN-driven expression of IL-15 in response to viral infection prepares memory CD8+ T cells for rapid division independently of antigen reexposure by transiently inducing cell-cycle progression via a pathway dependent on mTOR complex-1 (mTORC1). Moreover, exposure to IL-15 allowed more rapid division of memory CD8+ T cells following antigen encounter and enhanced their protective capacity against viral infection. Together, these data reveal that inflammatory IL-15 promotes optimal responses by memory CD8+ T cells.

To identify risk factors for venous thromboembolism (VTE) during the rehabilitation phase of spinal cord injury.A comprehensive review of the charts of all patients admitted between 1992 and 1995 for rehabilitation after spinal cord injury. Only records including evidence for objective testing for thromboembolism were included. The analysis variables included type and location of spinal cord injury, American Spinal Injury Association classification, concomitant injuries, surgical procedures, complications, preexisting illnesses, and use of antithrombotic prophylaxis. Hierarchically optimal classification tree analysis was employed to develop a nonlinear model for predicting deep vein thrombosis.The analysis sample consisted of these 243 persons with acute spinal cord injury, 51 of whom had venous thromboembolism, and eight of whom died. A three-variable model emerged that identified patient groups differing in relative likelihood of experiencing deep vein thrombosis. The highest likelihood group were patients with cancer over the age of 35 yrs. Women between the ages of 36 and 58 yrs without cancer were also at increased risk, as were cancer-free men with flaccid paralysis.Venous thromboembolism is more likely to develop in spinal cord injury patients who are older, obese, and have flaccid paralysis or cancer. These patients should receive vigorous prophylaxis against venous thromboembolism.

Effective CD8 T cell responses are vital for the control of chronic viral infections. Many factors of the host immune response contribute to the maintenance of effector CD8 T cell responses versus CD8 T cell exhaustion during chronic infection. Specific MHC alleles and the degree of MHC heterogeneity are associated with enhanced CD8 T cell function and viral control during human chronic infection. However, it is currently unclear to what extent host genetics influences the establishment of chronic viral infection. In order to examine the impact of MHC heterogeneity versus non-MHC host genetics on the development of chronic viral infection, an F1 cross of B10.D2 x B6 (D2B6F1) and BALB.B x BALB/c (BCF1) mice were infected with the clone-13 (Cl-13) strain of lymphocytic choriomeningitis virus (LCMV). Following chronic Cl-13 infection both H-2bxd D2B6F1 and BCF1 mice demonstrated increased viral control compared to homozygous mice. Strikingly, H-2bxd D2B6F1 mice on a C57BL genetic background exhibited mortality following Cl-13 infection. CD8 T cell depletion prevented mortality in Cl-13-infected D2B6F1 mice indicating that mortality was CD8 T-cell-dependent. D2B6F1 mice maintained more CD8 T cell effector cytokine production and exhibited reduced expression of the T cell exhaustion marker PD-1. In addition, D2B6F1 mice also induced a larger Th1 response than BCF1 mice and Cl-13-induced mortality in D2B6F1 mice was also dependent on CD4 T-cell-mediated IFN-γ production. Thus, following a chronic viral infection, increased functionality of the CD8 T cell response allowed for more rapid viral clearance at the cost of enhanced immunopathology dependent on both MHC diversity and the genetic background of the host., Author summary Chronic viral infections pose a serious healthcare concern resulting in substantial mortality worldwide. Chronic viral infections result from the inability of the immune system to eliminate the virus from the infected individual. The immune system’s inability to eradicate the invading pathogen is partially due to excessive regulation of the T cell response. However, host genetics have been associated with enhanced T cell function and viral control during chronic infection. Therefore, we sought to investigate the role of host genetic diversity on the T cell response during chronic viral infection in a murine model. We found that increasing MHC heterogeneity resulted in an increased T cell response and enhanced viral control. In addition, host genetic background differences allowed for induction of a distinct CD4 T cell subset, which was associated with reduced suppression of the CD8 T cell response and enhanced viral control. Thus, specific features of the host genetic background contribute to the size and quality of the T cell response and resulting viral control. This study identifies components of the T cell response that may provide a therapeutic target to enhance T-cell-mediated viral control during chronic infection.

There is no currently licensed vaccine for respiratory syncytial virus (RSV) despite being the leading cause of lower respiratory tract infections in children. Children previously immunized with a formalin-inactivated RSV (FI-RSV) vaccine exhibited enhanced respiratory disease following natural RSV infection. Subsequent studies in animal models have implicated roles for CD4 T cells, eosinophils and non-neutralizing antibodies in mediating enhanced respiratory disease. However, the underlying immunological mechanisms responsible for the enhanced respiratory disease and other disease manifestations associated with FI-RSV vaccine-enhanced disease remain unclear. We demonstrate for the first time that while CD4 T cells mediate all aspects of vaccine-enhanced disease, distinct CD4 T cell subsets orchestrate discrete and specific disease parameters. A Th2-biased immune response, but not eosinophils specifically, was required for airway hyperreactivity and mucus hypersecretion. In contrast, the Th1-associated cytokine TNF-α was necessary to mediate airway obstruction and weight loss. Our data demonstrate that individual disease manifestations associated with FI-RSV vaccine-enhanced disease are mediated by distinct subsets of CD4 T cells., Author Summary RSV is a significant healthcare burden and is the leading cause of bronchiolitis and pneumonia during childhood. The failure of the 1960's FI-RSV vaccine trial to not only elicit protection against RSV infection, but also provoke enhanced morbidity and mortality in vaccinees has significantly hampered development of new RSV vaccines for fear of disease potentiation. Therefore we sought to determine the specific immunological mechanisms that mediate FI-RSV VED to provide a framework to evaluate factors associated with disease exacerbation. Work presented herein demonstrate for the first time that individual disease manifestations associated with FI-RSV-immunization are mediated by distinct CD4 T cell subsets and not by eosinophils. Our results stress the need to evaluate multiple disease parameters for future RSV vaccine candidates. Failure to thoroughly assess the immune response and disease manifestations associated with new candidate vaccines may lead to undesired results in vaccine trials and further hinder future vaccine development.

Background Cellular bioenergetics (cellular respiration and accompanying ATP synthesis) is a highly sensitive biomarker of tissue injury and may be altered following infection. The status of cellular mitochondrial O2 consumption of the lung in pulmonary RSV infection is unknown. Methods In this study, lung fragments from RSV-infected BALB/c mice were evaluated for cellular O2 consumption, ATP content and caspase activity. The disease was induced by intranasal inoculation with the RSV strain A2 and lung specimens were analyzed on days 2–15 after inoculation. A phosphorescence O2 analyzer that measured dissolved O2 concentration as a function of time was used to monitor respiration. The caspase-3 substrate analogue N-acetyl-asp-glu-val-asp-7-amino-4-methylcoumarin (Ac-DEVD-AMC) was used to monitor intracellular caspases. Results O2 concentration declined linearly with time when measured in a sealed vial containing lung fragment and glucose as a respiratory substrate, revealing its zero-order kinetics. O2 consumption was inhibited by cyanide, confirming the oxidation occurred in the respiratory chain. Cellular respiration increased by 1.6-fold (p Conclusions Lung tissue bioenergetics is transiently enhanced in RSV infection. This energy burst, triggered by the virus or virus-induced inflammation, is an early biomarker of the disease and may be targeted for therapy.

The age-associated chronic thymus involution is interpreted to occur due to cytolytic depletion of thymic stromal tissue whose cells present altered self-peptides. Using simplified assumptions and based on morphometric data on thymic involution in man, the chance for a single protein to be altered is estimated to be in the range of 2-4 x 10(-6) per year. The corresponding mutation rate is compatible with that derived from both evolutionary and direct studies, thus supporting the proposed model.

Background Frozen section analysis has traditionally been used to confirm the identity of parathyroid tissue intraoperatively; however, it is time-consuming and costly and requires the excision of a significant portion of tissue. An intraoperative biopsy and analysis with a parathyroid hormone (PTH) assay is a possible alternative; however, this technique has not been perfected. Methods Two hundred twenty-three tissue specimens were collected prospectively from patients undergoing neck exploration. Each specimen was sampled intraoperatively using three different biopsy techniques: a fine-needle aspiration (FNA) with 10 passes of a needle (FNA10), a FNA with 20 passes of a needle (FNA20), and a tissue biopsy of approximately 1.0 mm 3 (BIOPSY). The PTH concentration of each sample was determined via the Elecsys® 1010 PTH immunoassay. The final tissue diagnosis was determined by histology or operative data. Results Parathyroid samples from all techniques had higher median PTH concentrations than nonparathyroid samples. However, the accuracies for the detection of parathyroid tissue varied markedly (PTH cutoff of 1000 pg/ml): the accuracies of the FNA10 and FNA20 were 71 and 80%, respectively, while the BIOPSY was 99% accurate. Conclusions This is the first prospective study evaluating multiple methods to diagnose parathyroid tissue intraoperatively using a rapid PTH assay. We conclusively show that the BIOPSY technique is 99% accurate for the diagnosis of parathyroid tissue, and therefore, should be the method of choice when the intraoperative confirmation of parathyroid tissue is needed.

Reduced fertility in men with spinal cord injury results from the inability to ejaculate and poor semen quality. Vibratory penile stimulation can induce ejaculation in many men with spinal cord injuries, but few studies have reported the effects of repeated vibratory stimulation on quantitative semen analysis tests. Fourteen spinal cord-injured males were enrolled in a prospective study to develop a quantitative profile of antegrade and retrograde ejaculate specimens collected during repeated vibratory stimulation. Antegrade specimens were obtained in 51 (84%) and retrograde specimens were obtained in all of the 61 trials (100%). Sperm were present in 70% and 64% of the antegrade and retrograde samples, respectively. Of the patients who underwent repeated vibratory stimulation attempts, three showed marked improvement in sperm counts and characteristics with repeated vibration and five showed normal fluctuation or no change in semen quality. Overall, there was a trend toward higher sperm counts in the antegrade samples (mean, 74.1 million) than in the retrograde samples (mean, 40 million), but no statistical significance was found. Mean sperm motility, sperm progressive motility, and percent normal sperm morphology were not significantly different between antegrade and retrograde specimens. Fructose and zinc, semen markers that reflect the presence of a true ejaculate, were present in all antegrade and retrograde specimens. In conclusion, (1) both antegrade and retrograde ejaculation occur with vibratory penile stimulation, with antegrade samples containing greater numbers of sperm but retrograde samples also having significant sperm counts, (2) repeated vibration may increase sperm concentration or may result in only minimal changes reflective of normal fluctuations, and (3) vibratory stimulation results in true ejaculation, as evidenced by semen markers.

Quality of life (QoL) is generally found to improve for renal transplant recipients, although some patients continue to experience health-related problems. It was within this context that we undertook our investigation which focused on identifying the factors predictive of QoL following kidney transplantation.The sample included 91 non-diabetic patients of which 69 provided 6-month data and 68 provided 12-month data. Three QoL questionnaires were administered to capture as many QoL dimensions as possible. Repeated measure analyses of variance with multiple post hoc comparisons of LS means was conducted to determine how QoL outcomes differed over time. Correlational analyses were performed on the 12-month dataset to determine which variables to include in the modeling process. Multiple stepwise regression with forward and backward entry were used in the prediction modeling.Essentially all patients experienced a significant improvement in QoL and the improvement occurred early and appeared to be sustained. Five separate prediction models were constructed, each including number of hospital days in first 6 months, employment, and social support.The similarity of the five models is of note. It is not necessarily these specific variables per se that predict QoL outcomes, but rather what they conceptually represent. These findings provide direction for interventions designed to enhance post-transplant QoL.

We urgently need to find a model for the aging processes and age-related diseases (cardiovascular; tumors), in order to be able to make a detailed appraisal of the many results obtained. An "evolutionary" theory of aging is proposed, in which the diminishing function of the thymus gland plays an important part.

As compared with quiescent lymphocytes, human thymocytes were found in the nucleoid sedimentation assay to be deficient in UV-induced DNA repair. Therefore, the age-related repair deficiency of human lymphocytes can result from the increased proportion of immature T cells occurring with age.

Acquisition of immunologic self-tolerance by clonal deletion in the thymus is concluded to be a critical process in immune aging and is found to be transiently protected from genetic instability by means of a feedback surveillance which functions at the expense of chronic thymic involution. In the immune control of mammalian aging, principles of Darwinian natural selection are suggested to work at the cellular level. During life, an immune-mediated stabilizing cell selection is displaced by a destabilizing one, thus changing physiological to pathological aging. In consequence, mammalian aging may be approached from an evolutionary point of view by both genetic cell variation and immune selection and be revealed correspondingly both as a stochastic and programmed process. Age-dependent diseases may be characterized as the consequence of localized pathological aging.

Human quiescent lymphocytes were lysed onto neutral sucrose gradients in order to sediment subsequently the nuclear DNA released within nucleoids. The position of nucleoids in the centrifuge tubes was detected fluorometrically by using the dye, ethidium bromide, and the height of the fluorescence peak was taken as a measure of DNA content. X-irradiation of lymphocytes, before their lysis, altered the DNA content of nucleoids and their sedimentation rate in accord with the view that (1) nuclear DNA is attached along its length at distances corresponding to 1.7 x 10(10) g/mol, and that (2) X-ray-induced double-strand breakage releases DNA fragments at random. Incubation at 37 degrees C of irradiated lymphocytes restored the amount of attached DNA as it would be expected from an intracellular repair process for DNA double-strand breaks.

The Royal District Nursing Service in Melbourne has been providing domiciliary nursing care to persons with HIV/AIDS since 1985. This paper outlines the development of the RDNS AIDS Nursing Program and gives an overview of the role of RDNS in management of HIV/AIDS in the community.

Highly purified human granulocytes isolated from continuous flow centrifugation leukapheresis concentrates by counterflow centrifugation-elutriation were stored at 4 °C in concentrations of 6 × 106 to 1 × 107 granulocytes per milliliter for up to 14 days. The in vitro physiological function assays of phagocytosis, oxygen consumption associated with phagocytosis, bacterial growth inhibition, chemotaxis, and five enzyme analyses indicated good storage survival for up to 4 days. Stored granulocytes separated from other blood cells have greater storage stability than granulocytes stored as leukapheresis concentrates. After 14 days of storage a small percentage of granulocytes still maintained all physiological functions, with the exception of chemotaxis. Of the five enzymes assayed, only the enzyme activity of leucine aminopeptidase decreased significantly by the 14th day of storage. The storage stability of each physiological function assayed decreased as follows: bacterial growth inhibition (most stable), phagocytosis, oxygen consumption, and chemotaxis (least stable).

Human unfractionated mononuclear leukocyte (UFMNL) preparations and T-cell-depleted (TCD) mononuclear leukocyte preparations, obtained from 400 ml of whole peripheral blood (PB) or from continuous-flow centrifugation leukapheresis (CFCL) concentrates, were cleanly separated by counterflow centrifugation-elutriation (CCE) into highly purified lymphocyte and monocyte populations, each with excellent recovery and function. Lymphocytes isolated from UFMNL preparations ranged in number from 1.88 × 10 8 to 2.22 × 10 9 with mean purities in excess of 93%. Lymphocytes isolated from TCD preparations yielded up to 1.90 × 10 9 lymphocytes devoid of all detectable sheep RBC receptor-bearing cells with mean purity and recovery in excess of 93 and 90%, respectively. Monocytes negatively selected from UFMNL preparations obtained from whole PB resulted in yields of 1.08 × 10 8 monocytes with 92% purity and 94% recovery; UFMNL obtained from CFCL concentrates resulted in yields of 4.22 × 10 8 monocytes with 92% purity. TCD preparations obtained from PB yielded 7.19 × 10 7 monocytes in 95% purity and 94% recovery, and from CFCL concentrates, 6.21 × 10 8 monocytes in 94% purity and 79% recovery. Recovered lymphocytes and monocytes have excellent overall viability, and the purified monocytes have normal phagocytic and chemotactic functional capabilities. Counterflow centrifugation-elutriation (CCE) is a valuable technique for obtaining monocyte and monocyte-depleted (i.e., lymphocyte) cell preparations that are negatively selected in high yield, with excellent purity and viability.

A long-time culture regime is tested for the production of lymphokine-activated killer cells from human lymphocytes, which combines the advantage of cell expansion with an increased cytotoxicity. The concentrations of IL-2 used are relatively low.

The superhelical properties of nuclear DNA from malignant cells of human origin (leukemic cell lines Reh and D-562, mesothelioma cell line, and peripheral blood monocytes from acute monocytic leukemia) were investigated in neutral sucrose gradients with ethidium bromide. As compared with the universal superhelical density of nuclear DNA from normal cells, the DNA released from malignant cells showed a substantially higher negative superhelix content, equivalent to an increased deficiency in right-handed DNA duplex turns. In regard to analogous results obtained from other malignant cell systems, the extremely underwound structural state of nuclear DNA appears to be a feature common to malignant cells.

Superhelical density and incision capacity for UV light-induced damage have been studied in nuclear DNA of human peripheral lymphocytes as a function of donor age. With advancing age between 20 and 90 years the content of negative superhelical turns increases, whereas the ability to incise UV lesions is impaired. These data may be suggestive of an immature lymphoid cell state arising in aging.

Beyond a donor age (a) of 22 years, the DNA incision ability in human peripheral lymphocytes was found to decrease according to 1/a2.5. Other lymphocyte functions linked to the differentiation program may follow this age-dependency. First results show that interleukin-2 is able to compensate, at least partly, the age-dependent deficiency of immune response.

In contrast to earlier studies, the presence of a stimulating capacity of cultured leukemic REH cells in a mixed lymphocyte culture is demonstrated. Supernatants of REH cells in a 1:10 final dilution were found to inhibit lymphocyte proliferation by about 30%. Mycoplasma contamination may introduce serious artefacts in all experimental systems which detect lymphocyte proliferation by means of incorporation of 3H-thymidine.

Characteristic changes of DNA winding and repair are detected in human peripheral lymphocytes with advancing donor age between 20 and 90 years, suggesting a progressively undifferentiated cell state. From a model approach it appears likely that the existence of immature lymphocytes favors aging as a general dysdifferentiation of somatic cells.

Normal human monocytes were negatively selected from leucapheresis cell suspensions by countercurrent centrifugation–elutriation in high yield with a mean purity of 93.5%. The combination of the novel methods of negative cell selection and suspension cell culture has provided the opportunity to study serially over several days the morphologic and functional changes of monocytes from a single donor as they matured in culture to typical macrophages. Human monocytes nearly double in size during the first week of culture, experiencing near daily increases in cell volume. This was associated with changes in the ultrastructure of these cells, including the development of numerous small knob-like projections on the cell membrane and the proliferation of microtubules and filamentous Structures within the cell cytoplasm during the first 6 days of culture. Peroxidase activity declined during the first 4 days of culture, whereas 5′-nucleotidase activity was acquired during the first 48 h of culture. Lysozyme activity in the cultures increased from day 2 to day 6 of culture. The phagocytic capacity of monocytes for IgG-coated erythrocytes increased dramatically during the first week of culture, but the cytotoxic capability of monocytes against similar targets in an antibody-dependent cytotoxicity assay declined to nearly hall of base-line levels by day 2 of culture and remained at this diminished level during subsequent days in culture.

From the X-irradiation doses needed to release supercoiling from nuclear DNA the mean size of its looped domains was calculated as being only about a half that in leukemic lymphoblasts as in normal or phytohemagglutinin-stimulated lymphocytes.

Human visually evoked cortical potentials (VEPs) were elicited by the onset of movement of grating patterns (test stimuli). The most prominent VEP waves N2 and P2 increased with accelerating test stimulus velocity (within a range of 0.2-4 deg/s). Two kinds of rums were presented. Reference runs involved only test stimuli and stationary pattern periods between them. In test runs moving gratings were additionally presented between the test stimuli effecting a stable level of movement adaptation. The additional movement stimulation reduced the VEP amplitudes throughout. The relative amplitude reduction was nearly constant for all test stimulus velocities if a fixed velocity of the adaptation stimulus was used. When the adaptation stimulus velocity was raised (within a range of 0.1-4 deg/s), the relative VEP amplitude became smaller. The results support the additive model of velocity coding in human occipital cortex rather than the substitutive model.

Knowledge referring to the kind of velocity coding in human cortex can be obtained either by psychophysical methods or by recording of visually evoked cortical potentials (VEPs). With the help of adaptation experiments it is possible to distinguish between the additive and substitutive coding principle. VEPs were evoked by the onset of movement of a grating pattern (test stimulus). The most prominent waves N2 and P2 increased with accelerating test stimulus velocity within a range of 0.2-4 deg/s (see strong curve in Fig. 6a and 7a). Moving gratings (adaptation stimuli) were additionally presented in corresponding runs during the intervals between the test stimuli to attain a stable level of movement adaptation. The additional movement stimulation reduced the VEP amplitudes in general (see weak lines in Figs. 6a, 7a and relative amplitudes in Figs. 6b, 7b). When the adaptation stimulus velocity was raised within a range of 0.1-4 deg/s (see values of the abscissas in Figs. 6 and 7 indicated by symbols with arrow) the relative VEP amplitude became smaller (decrease from curve to x). This result points to the validity of the additive coding principle. The different behavior of the delta-curve supports the hypothesis of a second channel for velocities faster than 1 deg/s.

An increased sedimentation rate of nucleoids can be monitored already 3 hours after mitotic stimulation of human peripheral lymphocytes. Additional folding of DNA onto the nuclear matrix is shown to be causally involved in this effect. The temporal attachments are likely associated with initiation of transcription.

A molecular mechanism is proposed by which physical, chemical and viral cancerogens together with DNA topoisomerases of type I effect an increase in the topological winding deficiency of nuclear DNA inside living cells. The resulting conformational changes of DNA, as monitored by an abnormally high negative superhelical density, could be of general relevance in the expression of the malignant cell phenotype.

24 hour blood-sugar profiles were obtained in 25 diabetic out-patients. The samples were obtained by the patients themselves even when at work, the glucose-level measurements in the laboratory. The relative differences between duplicate values was less than 10%. The results obtained made it possible to alter the treatment regimen so that, on average, there was a marked reduction in blood-sugar variations during the day.

Human granulocytes were isolated from leukapheresis blood by counterflow centrifugation-elutriation using the Beckman J21B centrifuge and JE6 rotor. Attempts to increase the absolute number of human granulocytes recovered by a single- or double-chamber procedure revealed the physical limitations of the chamber capacity in granulocyte isolation. To improve the absolute number of granulocytes from leukapheresis blood, an enlarged separation chamber and accompanying rotor were fabricated. The enlarged chamger has three times the capacity of the standard 4.5-ml Beckman chamber. Aside from increased yield of granulocytes, the 13.2-ml chamber affords a 25% saving in isolation time compared to the Beckman chamber for isolation of a comparable number of cells. In vitro analysis of the isolated granulocytes for viability, latex bead ingestion, and chemotactic response indicate no apparent loss of granulocyte function as a result of the isolation procedure.