Specific serine proteases selectively damage KCNH2 (hERG1) potassium channels and I(Kr).
KCNH2 (hERG1) encodes the alpha-subunit proteins for the rapidly activating delayed rectifier K+ current (I(Kr)), a major K+ current for cardiac myocyte repolarization. In isolated myocytes I(Kr) frequently is small in amplitude or absent, yet KCNH2 channels and I(Kr) are targets for drug block or mutations to cause long QT syndrome. We hypothesized that KCNH2 channels and I(Kr) are uniquely sensitive to enzymatic damage. To test this hypothesis, we studied heterologously expressed K+, Na+, and L-type Ca2+ channels, and in ventricular myocytes I(Kr), slowly activating delayed rectifier K+ current (I(Ks)), and inward rectifier K+ current (I(K1)), by using electrophysiological and biochemical methods. 1) Specific exogenous serine proteases (protease XIV, XXIV, or proteinase K) selectively degraded KCNH2 current (I(KCNH2)) and its mature channel protein without damaging cell integrity and with minimal effects on the other channel currents; 2) immature KCNH2 channel protein remained intact; 3) smaller molecular mass KCNH2 degradation products appeared; 4) protease XXIV selectively abolished I(Kr); and 5) reculturing HEK-293 cells after protease exposure resulted in the gradual recovery of I(KCNH2) and its mature channel protein over several hours. Thus the channel protein for I(KCNH2) and I(Kr) is uniquely sensitive to proteolysis. Analysis of the degradation products suggests selective proteolysis within the S5-pore extracellular linker, which is structurally unique among Kv channels. These data provide 1) a new mechanism to account for low I(Kr) density in some isolated myocytes, 2) evidence that most complexly glycosylated KCNH2 channel protein is in the plasma membrane, and 3) new insight into the rate of biogenesis of KCNH2 channel protein within cells.


[The influence of obesity on early results and quality of life of patients after chosen operations performed by laparoscopic or open methods].
Obesity usually depreciates early results of abdominal operations performed in an open way, but not always after laparoscopic operations. Aim of the study is to compare early results and quality of life after most common abdominal operations (cholecystectomy and appendectomy) performed on obese and non-obese patients. 166 patients who were operated in academic teaching hospital, were analyzed in groups: cholecystectomy - laparoscopic (74) and open (30), appendectomy - laparoscopic (30) and open (32). Every group was divided into subgroups with normal body mass, obesity I obesity II and was assessed retrospectively and investigated by Gastrointestinal Quality of Life Index (GQLI) up to 5 months after intervention by registered nurse specialized in surgery. Number and gravity of complications were assessed as well as hospital stay and convalescence time. There were 7/19 and 4/13 complications after laparoscopic vs 8/12 and 7/19 after open cholecystectomies and appendectomies respectively in groups with normal body weight (BMI 18.5-24.9kg/m2), and 28/56 and 8/20 complications after laparoscopic vs 14/18 and 9/12 after open cholecystectomies and appendectomies respectively in obese groups (above 25 kg/m2). One conversion to open surgery occured in patient with obesity I for laparoscopic cholecystectomy and one in normal body mass patient for laparoscopic appendectomy. There was one case of common bile duct lesion in obese patient during laparoscopic intervention and two cases of intestinal obstruction after open cholecystectomy in obese patients. Mean hospital stay time was 3.2 and 3.8 days after laparoscopic operations vs 5.4 and 5.4 days after open ones respectively. Mean convalescence time was 14 and 14 days after laparoscopic operations vs 21 and 14 days after open cholecystectomy and appendectomy respectively. Quality of life was significantly better after laparoscopic than open cholecystectomy in obese patients (128.4 +/- 12.7 vs 120.6 +/- 12.2 respectively) but significantly worse after laparoscopic than open appendectomy (121 +/- 15.2 vs 133.8 +/- 12.9 respectively). In groups with normal body mass differences were not significant. There is the influence of obesity on number and burden of postoperative complications after laparoscopic and open operations, however laparoscopic access seems more effective. Although hospital stay time was shorter after both types of laparoscopic operations, convalescence time was shorter only after laparoscopic cholecystectomy. The choice of type of operation (open or laparoscopic) influences the quality of life of obese patients but not on those with normal body mass. Although these conclusions seem rather clear, verification of them on larger group of patients is mandatory.


Disruption and maintenance of skilled visual search as a function of degree of consistency.
The present experiment was conducted to investigate the effects of varying degrees of task consistency on the performance and maintenance of skill in a semantic-category visual search task. Four groups of participants first received 6000 trials of consistent mapping (CM) training on two different categories. The participants then performed 4000 trials in which one of the previously trained categories remained 100% consistent, whereas the other previously trained category became either 100%, 67%, 50%, or 33% consistent. This second phase of the experiment allowed for the examination of disruption of the search skill as a function of degree of consistency. Subsequent to the degree of consistency manipulation, 100% consistency was restored and participants performed another 4200 CM trials. Results indicate that performance was disrupted by inconsistency and that disruption increased as consistency decreased. On the return of task consistency, performance improved rapidly to predisruption levels, though some performance disruption was evident. Theoretical and practical implications are discussed.


Endoscopic Management of Iatrogenic Colon Perforation.
Colon perforations are difficult to resolve because they occur unexpectedly and infrequently. If the clinician is unprepared or lacks training in dealing with perforations, the clinical prognosis will be affected, which can lead to legal issues. We describe here the proper approach to the management of perforations, including deciding on endoscopic or surgical treatment, selection of endoscopic devices, endoscopic closure procedures, and general management of perforations that occur during diagnostic or therapeutic colonoscopy.


Error reduction with routine checklist use during deep brain stimulation surgery.
The use of checklists to reduce error rates in procedural literature has led our group to employ this strategy during deep brain stimulation (DBS) surgery. We sought to examine the improvement in the number of errors made during DBS surgery after long-term use of a checklist. Our checklist has been used for all DBS cases at our institution since the beginning of this study's enrollment in 2008. The number of cases in which errors were detected after 1 year of routine use (group B, n = 11) was compared in one cohort of DBS subjects to that of an earlier cohort of patients (group A, n = 17), which underwent DBS exactly 1 year prior. Eleven of the 14 cases where major errors were detected occurred in group A; 6 of the 9 cases where only minor errors were detected were also in group A; of the patients without any error, all 5 were in group B. We found a significant difference in these proportions between group A and group B [χ(2)(2) = 9.73; p < 0.008]. After 1 year of checklist use, the total number of major and minor errors made was reduced, indicating an improvement in error rate after long-term routine incorporation of this checklist.


Inverse correlation between testosterone and ventricle ejection fraction, hemodynamics and exercise capacity in heart failure patients with erectile dysfunction.
Neurohormonal activation and abnormalities in growth hormone and testosterone concentrations have been reported in heart failure (HF). Erectile dysfunction(ED) is common in these patients and contributes to a low quality of life. No data are known regarding the correlation between testosterone and hemodynamics, exercise capacity and cardiac function in HF patients with ED, a marker of endothelial dysfunction. The aim of this study was to correlate testosterone levels with cardiac function, hemodynamic and exercise capacity in HF patients with ED. Fifteen HF patients underwent a six-minute treadmill cardiopulmonary walking test (6'CWT) and, ten minutes later, a maximum cardiopulmonary exercise test. Also, testosterone and other hormones were determined at rest. Among hemodynamic variables only diastolic blood pressure on 6'CWT was correlated with testosterone levels(r=-0.66, p=0.007). The variables on exercise tests, VE/VCO2 slope and oxygen consumption did not show any correlation, except the distance at 6'CWT (r=0.50, p=0,047). Right and left ventricle ejection fraction showed inverse correlation with testosterone (r=-0.55, p=0.03 and r=-0.69, p=0.004 respectively). Testosterone levels correlated directly with distance at six-minute cardiopulmonary walk test and inversely with diastolic blood pressure, right and left ventricle ejection fraction in heart failure patients with erectile dysfunction. Further elucidation of mechanisms as regards testosterone action in these patients is warranted.


Management of infantile hemangiomas : current and potential pharmacotherapeutic approaches.
Infantile hemangiomas (IH), benign vascular neoplasms, are the most common tumors of infancy and childhood. Most IH are medically insignificant; however, a proportion will require treatment because of interference with vital structures, threat of significant disfigurement, ulceration, or bleeding. This article reviews current and potential pharmacotherapeutic approaches to the treatment of IH. While corticosteroids have long been considered the mainstay of medical therapy for IH, several new treatments have recently emerged, the most promising of which is oral propranolol. Topical timolol and imiquimod are additional new therapies that may also prove to be effective, particularly for the treatment of superficial IH.


Classification of Pulmonary Nodular Findings based on Characterization of Change using Radiology Reports.
Radiology reports contain descriptions of radiological observations followed by diagnosis and follow up recommendations, transcribed by radiologists while reading medical images. One of the most challenging tasks in a radiology workflow is to extract, characterize and structure such content to be able to pair each observation with an appropriate action. This requires classification of the findings based on the provided characterization. In most clinical setups, this is done manually, which is tedious, time-consuming and prone to human error yet of great importance as various types of findings in the reports require different follow-up decision supports and draw different levels of attention. In this work, we present a framework for detection and classification of change characteristics of pulmonary nodular findings in radiology reports. We combine a pre-trained word embedding model with a deep learning based sentence encoder. To overcome the challenge of access to limited labeled data for training, we apply Siamese network with pairwise inputs, which enforces the similarities between findings under the same category. The proposed multitask neural network classifier was evaluated and compared against state-of-the-art approaches and demonstrated promising performance.


CD2 Immunobiology.
The glycoprotein CD2 is a costimulatory receptor expressed mainly on T and NK cells that binds to LFA3, a cell surface protein expressed on e.g., antigen-presenting cells. CD2 has an important role in the formation and organization of the immunological synapse that is formed between T cells and antigen-presenting cells upon cell-cell conjugation and associated intracellular signaling. CD2 expression is upregulated on memory T cells as well as activated T cells and plays an important role in activation of memory T cells despite the coexistence of several other costimulatory pathways. Anti-CD2 monoclonal antibodies have been shown to induce immune modulatory effects in vitro and clinical studies have proven the safety and efficacy of CD2-targeting biologics. Investigators have highlighted that the lack of attention to the CD2/LFA3 costimulatory pathway is a missed opportunity. Overall, CD2 is an attractive target for monoclonal antibodies intended for treatment of pathologies characterized by undesired T cell activation and offers an avenue to more selectively target memory T cells while favoring immune regulation.


The Effect of Environmental Performance on Employment: Evidence from China's Manufacturing Industries.
This research aims to explore the interaction between environmental performance and employment China's manufacturing industries. Based on the environmental performance of 32 industries in China's manufacturing industry during 2006-2015, a panel vector autoregressive model was constructed to study the interaction between industry output and employment in clean industries and dirty industries. The dynamic impact and internal transmission mechanism between environmental performance is analyzed. The study found that in the early stage, due to the reduction of production scale, there was a weak and short-term negative correlation effect on employment, and the mutual promotion relationship between economic benefits and employment was unsustainable. In return, employment affects environmental performance, but the effect differs due to the different forms of environmental performance. For dirty industries, the impact of environmental performance on employment through technical effects is more significant and, thus, a win-win situation of ecological environment and employment stability will be achieved. This research has practical significance regarding how to scientifically and effectively carry out environmental regulation and green management.


Changes of The Uterine Tissue in Rats with Polycystic Ovary Syndrome Induced by Estradiol Valerate.
Polycystic ovary syndrome (PCOS) is one of the most common hormonal disorders that can lead to irregular menstrual cycles and hyperandrogenism. Reduced levels of progesterone and increased estrogen in these women can perpetually stimulate the endometrial tissue of the uterus. In this study, we assess the effect of PCOS induction by estradiol valerate (EV) in a rat model. In this experimental study, adult female Wistar rats that weighed approximately 200 g were divided into control, sham, and experimental groups (n=6 per group). The experimental group received subcutaneous injections of 2 mg EV for induction of PCOS. We confirmed the presence of PCOS in the experimental group rats. Rats from all groups were subsequently killed, after which their uteri were removed and fixed for histological and cytological analyses. The uterine tissue sections were stained with hematoxylin and eosin (H&E) and iron hematoxylin (iron-H). We examined epithelium height, thickness of the uterus wall, and frequency of the mitotic cells. The data were assessed at α=0.05. Uterine tissue findings from the experimental group showed significant increases in the height of the uterus luminal epithelium, the thickness of the uterus wall, and the frequency of eosinophils in the endometrial stroma. We observed an increased frequency of mitotic cells in the experimental group in both luminal and glandular epithelia of the uterus. An increased rate of the glandular epithelium region was noticeable and significant. Induction of PCOS by EV could change the proliferation rate in the endo- metrial tissue of the uterus.


Thermal treatment decreases resistance to osimertinib in non-small cell lung cancer through the EGFR/PI3K/AKT pathway.
Osimertinib (OSI) resistance commonly occurs during the treatment of non-small-cell lung cancer (NSCLC). This study aims to investigate whether the thermal effects of radiofrequency ablation (RFA) can increase the sensitivity of OSI-resistant NSCLC to OSI treatment and whether OSI effectively inhibits the recurrence of OSI-resistant NSCLC following RFA treatment and improve survival of NSCLC patients. In vitro, OSI-resistant NCI-H1975 (NCI-H1975/OSIR) cells and thermotolerant NCI-H1975/OSIR (NCI-H1975/OSIR-a-h) cells were established using human NSCLC cell line NCI-H1975. Cell viability, apoptosis, sensitivity to OSI, threonine-methionine amino acid substitution at position 790 (T790M) mutation levels, and protein expression of epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), hypoxia-inducible factor-1 alpha (HIF-1a) were detected using different methods. In vivo, a nude mouse model of metastatic human lung cancer was developed and subjected to RFA treatment. The tumor growth, apoptosis, sensitivity to OSI, expression of EGFR/PI3K/AKT/HIF-1a, and CD34 levels were detected in the micrometastases of the transition zone (TZ) around the central ablation zone, and the reference zone (RZ) far from central ablation zone. NCI-H1975/OSIR and thermotolerant NCI-H1975/OSIR cell models were successfully established. Thermotolerant NCI-H1975/OSIR cells show higher sensitivity to OSI than NCI-H1975/OSIR cells and NCI-H1975 cells. OSI treatment can inhibit the EGFR/PI3K/AKT pathway and induce apoptosis in both NCI-H1975 cells and thermotolerant NCI-H1975/OSIR cells, but not in NCI-H1975/OSIR cells. In vivo, RFA treatment increases sensitivity to OSI in NCI-H1975/OSIR cell micrometastases in the TZ but not in the RZ. OSI intervention effectively inhibits the over-proliferation of micrometastases and activation of the EGFR/PI3K/AKT pathway, and induces apoptosis of micrometastases in the TZ, but shows little effects on the micrometastases in the RZ. The thermal effects can increase the sensitivity of OSI-resistant NSCLC cells to OSI through the EGFR/PI3K/AKT/HIF-1a signaling pathway, indicating that RFA combined with OSI might be a clinically effective and comprehensive therapy for the treatment of OSI-resistant NSCLC.


Polycistronic effects of catabolite repression on the lac operon.
The catabolite repression caused by glucose and glucose-6-phosphate has been studied for both beta-galactosidase and thiogalactoside transacetylase, the products of the operator proximal and distal cistrons of the lac operon, respectively. We find that both cistrons are affected coordinately by this form of repression. We also find that a single alteration at the lac promoter region is sufficient to abolish sensitivity to repression of both cistrons. From this, we conclude that there is only one target site for catabolite repression in the lac operon.


Efficacy of goserelin plus anastrozole in premenopausal women with advanced or recurrent breast cancer refractory to an LH-RH analogue with tamoxifen: results of the JMTO BC08-01 phase II trial.
The aim of the present study was to assess the efficacy and tolerability of a luteinizing hormone-releasing hormone (LH-RH) analogue plus an aromatase inhibitor following failure to respond to standard LH-RH analogue plus tamoxifen (TAM) in premenopausal patients. Premenopausal women with estrogen receptor (ER)-positive and/or progesterone-receptor positive, advanced or recurrent breast cancer refractory to an LH-RH analogue plus TAM received goserelin (GOS) in conjunction with anastrozole (ANA). The primary endpoint was the objective response rate (ORR). Secondary endpoints included progression-free survival (PFS), overall survival (OS), clinical benefit rate (CBR) and safety. Between September 2008 and November 2010, 37 patients were enrolled. Thirty-five patients (94.6%) had ER-positive tumors, and 36 (97.3%) had human epidermal growth factor receptor (HER) 2-negative tumors. Thirty-six (97.3%) had measurable lesions and 1 (2.7%) had only bone metastasis. The ORR was 18.9% [95% confidence interval (CI), 8.0-35.2%], the CBR was 62.2% (95% CI, 44.8-77.5%) and the median PFS was 7.3 months. Eight patients had adverse drug reactions but none resulted in discontinuation of treatment. GOS plus ANA is a safe effective treatment for premenopausal women with hormone receptor-positive, recurrent or advanced breast cancer. The treatment may become viable treatment in the future, particularly when TAM is ineffective or contraindicated. Further studies and discussion are warranted.


Diagnosis and treatment of prostate cancer.
In the United States, prostate cancer is the most common solid tumor malignancy in men and second to lung cancer as the leading cause of cancer deaths in this group. Even though prostate cancer is responsible for 40,000 deaths per year, screening programs are a matter of controversy because scientific evidence is lacking that early detection decreases morbidity and mortality. Furthermore, treatment decisions are difficult to make because of the generally indolent nature of prostate cancer and because it tends to occur in older men who often have multiple, competing medical illnesses. Depending on the specific situation, radical prostatectomy, radiotherapy or watchful waiting (observation) will be the most appropriate management option. In general, localized cancer is best treated with surgical removal of the prostate gland or radiotherapy. Hormone deprivation therapy is the primary method of controlling metastatic prostate cancer. At present, chemotherapy cannot cure disseminated prostate cancer. Watchful waiting is a reasonable management alternative for prostate cancer in an older patient or a patient with other serious illnesses.


Airway closure, atelectasis and gas exchange during anaesthesia.
Pulmonary gas exchange is regularly impaired during general anaesthesia with mechanical ventilation. This results in decreased oxygenation of blood. Major causes are collapse of lung tissue (atelectasis) and airway closure. Collapsed lung tissue is present in 90% of all subjects, both during spontaneous breathing and after muscle paralysis, and whether intravenous or inhalational anaesthetics are used. Airway closure is also common and increases in magnitude with increasing age of the patient. There are correlation between the amount of atelectasis and pulmonary shunt and between airway closure and perfusion of poorly ventilated lung regions (low VA/Q). Atelectasis and airway closure explain as much as 74% of gas exchange impairment in routine anaesthesia. A major cause of atelectasis is the pre-oxygenation during induction of anaesthesia. Lowering the inspired O2 concentration to 80% suffices to avoid almost all atelectasis. Airway closure and low VA/Q can only be prevented by raising the FRC level by PEEP or by other means.


Biodemography of a long-lived tephritid: reproduction and longevity in a large cohort of female Mexican fruit flies, Anastrepha ludens.
Age of sexual maturity, daily and lifetime reproductive rates, and life span were recorded in a laboratory cohort of Mexican fruit flies consisting of over 1100 females maintained individually. The results revealed that, relative to the medfly, the Mexfly is slower maturing (14 vs 17 days), more fecund (1400 vs 650-1100 eggs/female), and longer lived (50 vs 35 days). The results reinforced the generality of several earlier findings on the medfly including the deceleration of mortality at older ages and the weakness of the correlation between the rate of egg laying at early ages and both subsequent reproduction and remaining longevity. Discussion includes perspectives on the role of artificial selection in shaping the demographic traits of the mass-reared strain of Mexfly used in this study, as well as the overall significance of large scale biodemographic studies in understanding aging and longevity.


NOSTRIN functions as a homotrimeric adaptor protein facilitating internalization of eNOS.
Intracellular trafficking of endothelial nitric oxide synthase (eNOS) between different compartments is incompletely understood. Recently, we described a novel eNOS-interacting protein, NOSTRIN, which upon overexpression drives eNOS away from the plasma membrane towards intracellular compartments. Sequence similarity of NOSTRIN and pacsins/syndapins suggested a role for NOSTRIN in endocytosis. Accordingly, we show here that NOSTRIN interacts with the large GTPase dynamin and the actin nucleation promoting factor N-WASP by means of its SH3 domain, which also represents the docking site for eNOS. Via a coiled-coil region in the C-terminal portion of the protein, NOSTRIN oligomerizes, mainly forming trimers, which would allow simultaneous interaction with multiple binding partners of the SH3 domain. Consistent with this notion, expression of dynamin-2-GFP in CHO cells stably expressing eNOS (CHO-eNOS) results in recruitment of eNOS to dynamin-positive structures, only when NOSTRIN is present as well. Similarly, when N-WASP-GFP and NOSTRIN are co-expressed in CHO-eNOS cells, both proteins strongly co-localize with eNOS and are recruited to structures running along actin filaments. If, however, the actin cytoskeleton is depolymerized by cytochalasin D, NOSTRIN and eNOS are associated with extended structures in the cell periphery, possibly being unable to leave the plasma membrane. Together, these results indicate that NOSTRIN may facilitate endocytosis of eNOS by coordinating the function of dynamin and N-WASP.


Endovascular versus conventional open surgical repair for thoracoabdominal aortic aneurysms.
Thoracoabdominal aortic aneurysms (TAAAs) are a life-threatening condition which remain difficult to treat. Endovascular and open surgical repair (OSR) provide treatment options for patients, however, due to the lack of clinical trials comparing these, the optimum treatment option is unknown. To assess the effectiveness and safety of endovascular repair versus conventional OSR for the treatment of TAAAs. The Cochrane Vascular Information Specialist searched the Cochrane Vascular Specialised Register, CENTRAL, MEDLINE, Embase, CINAHL and AMED databases and World Health Organization International Clinical Trials Registry Platform and ClinicalTrials.gov trials registers to 26 April 2021. We also searched references of relevant articles retrieved from the electronic search for additional citations. We considered all published and unpublished randomised controlled trials (RCTs) and controlled clinical trials (CCTs) comparing endovascular repair to OSR for TAAAs for inclusion in the review. The main outcomes of interest were prevention of aneurysm rupture (participants without aneurysm rupture up to 5 years from intervention), aneurysm-related mortality (30 days and 12 months), all-cause mortality, spinal cord ischaemia (paraplegia, paraparesis), visceral arterial branch compromise causing mesenteric ischaemia or renal failure, and rate of reintervention. Two review authors independently screened all titles and abstracts identified from the searches to identify those that met the inclusion criteria. We planned to undertake data collection, risk of bias assessment, and analysis in accordance with Cochrane recommendations. We planned to assess the certainty of the evidence using GRADE. No RCTs or CCTs met the inclusion criteria for this review. Due to the lack of RCTs or CCTs, we were unable to determine the safety and effectiveness of endovascular compared to OSR in patients with TAAAs and are unable to provide any evidence on the optimal surgical intervention for this cohort of patients. High-quality RCTs or CCTs addressing this objective are necessary, however conducting such studies will be logistically and ethically challenging for this life-threatening disease.


Association between magnesium intake and cognition in US older adults: National Health and Nutrition Examination Survey (NHANES) 2011 to 2014.
Identifying nutrition- and modifiable lifestyle-based risk factors for cognitive decline and dementia may contribute future primary prevention strategies. This study aimed to evaluate the associations between magnesium intake and cognition in older adults in the United States. Based on the National Health and Nutrition Survey (NHANES) between 2011 and 2014, the study included 2508 participants aged 60 years and older. Linear regression models were used to examine the association of total magnesium intake with cognition. After adjusted demographic and other confounding factors, intakes of energy and total calcium, and serum vitamin D level, higher intake of total magnesium was independently associated with 0.15 higher global cognitive z-score (95% confidence interval, 0.02 to 0.28 for highest vs. lowest quartile, P trend = .037). The positive association of total magnesium intake with global cognition was primarily presented among women, non-Hispanic Whites, and those with sufficient serum vitamin D levels (≥50 nmol/L), although interactions were not significant. There were no clear linear associations for global cognition with serum vitamin D level. Our findings suggest that high magnesium intake alone may improve cognition in older adults, particularly among non-Hispanic Whites and subjects with sufficient levels of serum vitamin D. Further studies are needed to confirm the findings.


IL-8 release of HL-60 cells treated with electric currents of different wave forms.
Human promyelocytic leukaemia HL-60 cells which have been differentiated by DMSO to granulocytes were used to investigate the effect of different waveforms on the release of interleukine-8 (IL-8). The cells were prestimulated with 100 pM fMLP and subsequently treated for 15 min with different electrical fields and currents. Three hours later the release of IL-8 into the medium was determined by ELISA. Varying the frequency of the sinusoidal electrical current between 0 and 20 Hz resulted in 2 maxima of IL-8 release at 5 and 13 Hz. Prestimulated cells were treated with sine-, triangular-, and rectangular-waveforms at 5 Hz in the current intensity range of 0-3 mA/cm(2). For the three waveforms tested, the IL-8 release was enhanced 1.5 fold. Treatment of the cells with capacitively coupled electric fields of 5 Hz using field strengths between 0 and 10 V(eff)/cm had no effect on the release of IL-8. In comparison to the positive results after sine wave exposure alone, an exposure with sine wave current to which noise had been superimposed had no effect on the HL-60 cells. From these investigations it can be concluded that for electrical current treatment of prestimulated HL-60 cells the release of IL-8 does not depend on the waveform if the waveform information is not destroyed by superimposed noise.


Audiology in an ecological perspective--development of a conceptual framework.
The conceptual framework for describing an audiological communication handicap and its underlying processes is built in analogy to a biological, ecological system and uses the following primary concepts: interaction, internal milieu, preferendum (preferred communication features) and optimal resource utilization. From these primary concepts secondary concepts and processes are formulated: signal, message/dialogue and behavior are three levels of interaction; adjustment and release of mental resources are specifications for preferendum (influenced by contextual and social conditions); specificity and facilitation underlie optimal resource utilization. The internal environment contains afferent, central and efferent speech and language processing, cognitive and emotional processes. The main components of audiology and rehabilitation (impairment, disability, handicap) are deduced from these concepts and processes. Examples of studies based on this conceptual framework are presented briefly. These include an environmental test chamber where realistic acoustic environments and communication situations can be directly studied, and a rehabilitation programme based on tutorial education for structured interaction at the behaviour level between hearing-impaired and normal-hearing persons. A vibratory-based event localization system for deaf-blind is described which improves specificity and thereby mental resource utilization.


Effect of spatial separation and noise type on sentence recognition by Mandarin-speaking cochlear implant users.
To investigate the effects of spatial separation and noise type on sentence recognition by unilateral Mandarin-speaking cochlear implant (CI) users and normal-hearing (NH) listeners. Twenty-two unilateral Mandarin-speaking CI users and six NH listeners participated in this study. Speech reception thresholds were measured for three noise types (steady state noise, speech babble, and music). Sentences from the Mandarin Speech Perception test were presented directly in front of the listener (0°). Noise was presented from one of the five speaker locations: -90°, -45°, 0°, +45°, and +90°. Overall, CI performance was significantly poorer than NH performance for all spatial separation and noise type conditions. NH listeners performed best with music and poorest with steady noise. CI users performed best with steady noise, and poorest with babble. Performance was significantly affected by noise location and noise type. There was no significant difference in head shadow effects among the different noise types for CI users. Performance was much poorer in CI than in NH listeners for all noise types and spatial separations. Noise type differently affected unilateral CI users and NH listeners. The limited spectral resolution in CI users did not appear to affect head shadow.


Charge heterogeneity of monoclonal antibodies by multiplexed imaged capillary isoelectric focusing immunoassay with chemiluminescence detection.
Characterization of charge heterogeneity of recombinant monoclonal antibodies (mAbs) requires high throughput analytical methods to support clone selection and formulation screens. We applied the NanoPro technology to rapidly measure relative charge distribution of mAbs in early stage process development. The NanoPro is a multiplexed capillary-based isoelectric immunoassay with whole-column imaging detection. This assay offers specificity, speed and sensitivity advantages over conventional capillary isoelectric focusing (CIEF) platforms. After CIEF, charge variants are photochemically immobilized to the wall of a short coated capillary. Once immobilized, mAbs are probed using a secondary anti-IgG conjugated with horseradish peroxidase. After flushing away excess reagents, secondary antibodies bound to their targets are then detected by chemiluminescence upon incubation with peroxidase reactive substrates. Charge heterogeneity as determined by chemiluminescence was similar to that measured by conventional CIEF technology with absorbance detection for purified mAbs and contaminated mAbs derived directly from host cellular extract. Upon method optimization, the automated CIEF immunoassay was applied to several mAbs of varying isoelectric points, demonstrating the suitability of NanoPro as a rugged high-throughput product characterization tool. Furthermore, qualification of detection sensitivity, precision, and dynamic range are reported with discussion of its advantages as an alternative approach to rapidly characterize charge variants during process development of mAbs.


Thymopentin loaded microsphere preparation by w/o/w emulsion technique: in vitro/ex vivo evaluation.
Poly-D,L-lactide (PDLLA) and polylactide-co-glycolide (PLGA) microspheres containing thymopentin have been prepared by a water-in-oil-in-water-emulsion/solvent evaporation technique. The goal is to stabilize the active compound thymopentin, and to prolong its therapeutic activity, by embedding the drug in a polymeric matrix. The microspheres obtained have been characterized for their morphology and drug content. In-vitro dissolution tests have been performed on the microspheres. Results show that the type of polymer employed (PDLLA or PLGA) does not seem to affect microsphere morphology, while in-vitro dissolution profiles are greatly influenced by the composition of polymer matrix. Ex-vivo evaluation of PLGA microspheres performed on mouse thymocites shows that biological activity of Thymopentin is maintained after loading into PLGA microspheres.


Is virtual bronchoscopy useful for physicians practising in a district general hospital?
Virtual bronchoscopy software is now available to district general hospitals (DGHs). There is limited information on the clinical utility of virtual bronchoscopy and whether it offers any additional information over conventional axial computed tomography in the setting of a busy DGH chest unit. Virtual bronchoscopy and computed tomography findings were compared in all patients who had a virtual bronchoscopy study over a 12 month period. Eighteen consecutive patients had virtual bronchoscopy for a specific clinical indication over the study period. Additional information was conveyed by virtual bronchoscopy in five patients (in four patients the airways distal to an obstruction were better visualised thereby influencing decisions about airway stenting and in one patient the virtual bronchoscopy study showed an endobronchial lesion missed on computed tomography). In nine patients who were unfit for fibreoptic bronchoscopy (FOB) the radiologist was more confident in excluding an obstructive airway lesion. The main indication for performing a virtual bronchoscopy study was to rule out an obstructive airway lesion in patients who were unfit for FOB (n = 11). Virtual bronchoscopy is feasible and useful in the management of a few selected patients in a DGH chest unit. Virtual bronchoscopy may convey additional information over computed tomography when the distal airways need to be visualised and for discrete endoluminal lesions.


Efficacy of hydroxocobalamin for the treatment of acute cyanide poisoning in adult beagle dogs.
The efficacy of hydroxocobalamin for acute cyanide poisoning was compared with that of saline vehicle in dogs. Anesthetized adult beagle dogs were administered potassium cyanide (0.4 mg/kg/min, IV) until 3 min after the onset of apnea. Hydroxocobalamin (75 mg/kg [n = 19] or 150 mg/kg [n = 18], IV) or saline vehicle [n = 17] was then infused over 7.5 min while animals were ventilated with 100% oxygen, which was stopped after 15 min. In vehicle-treated animals cyanide produced deterioration that culminated in a moribund state requiring euthanasia within 4 h in 10 of 17 animals and in neurological deficits necessitating euthanasia within 2-4 d in an additional 4 animals (mortality rate 82%). Survival through 14 d was observed in 15 of 19 animals administered hydroxocobalamin 75 mg/kg (mortality rate 21%), and 18 of 18 administered hydroxocobalamin 150 mg/kg (mortality rate 0%). Hydroxocobalamin reversed cyanide toxicity and reduced mortality in a canine model.


Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov.
Previous DNA relatedness and enzyme electrophoretic mobility studies indicated heterogeneity among strains of Legionella pneumophila serogroups 1, 4, 5, and Lansing 3 (a new, as yet unnumbered serogroup). In this study 60 L. pneumophila strains were studied by DNA hybridization (hydroxyapatite method) to assess their genomic relatedness. These strains were also studied biochemically and serologically to determine whether they formed one or more phenotypic groups. DNA relatedness studies identified three groups. DNA group 1 contained the type strain Philadelphia 1 and strains from serogroups 1 through 14 of L. pneumophila. The average relatedness of DNA group 1 strains was 88% at 60 degrees C with 1.1% divergence in related sequences and 85% at 75 degrees C. DNA group 2 contained strain Los Angeles 1, the reference strain of serogroup 4, and strains of serogroups 1, 4, 5, and Lansing 3, an unnumbered serogroup. Average relatedness of DNA group 2 strains was 84% at 60 degrees C with 0.7% divergence and 87% at 75 degrees C. Reciprocal relatedness of DNA groups 1 and 2 was approximately 67% at 60 degrees C with 6.0% divergence and 48% at 75 degrees C. DNA group 3 strains were in serogroup 5. They were 98% related at 60 degrees C with 0.5% divergence and 97% related at 75 degrees C. Reciprocal relatedness of DNA group 3 and DNA group 1 was approximately 74% at 60 degrees C with 5.3% divergence and 43% at 75 degrees C, and reciprocal relatedness of DNA groups 3 and 2 was 66% at 60 degrees C with 5.7% divergence and 55% at 75 degrees C. The DNA groups could not be separated biochemically or serologically or by cell wall fatty acid and isoprenoid quinone composition. Three subspecies of L. pneumophila are proposed to accommodate the three DNA groups: L. pneumophila subsp. pneumophila subsp. nov. for DNA group 1, L. pneumophila subsp. fraseri subsp. nov. for DNA group 2, and pneumophila subsp. pascullei subsp. nov. for DNA group 3.


[Measures, motivation and influence of Yan Xishan's support for traditional Chinese medicine in the Republic of China].
In Shanxi Province, governed by Yan Xishan during the Republic of China, traditional Chinese medicine (TCM) is in a climate of relative tolerance on the whole, under its supporting policy. A TCM Improving Society was organized by Yan Xishan himself as president. Whereafter, a series of activities are carried out by the Society, such as the establishments of an Affiliated Hospital, a Learning School of TCM, and creation of Medical Magazineetc., all of which was supported by Yan through providing financial resources and manpower. The reasons for Yan's such supports, in addition to his deeply influenced by traditional Chinese culture and his recognition on the therapeutic effects of TCM, might lie in the account of the practical reality of economy and people's livelihood in Shanxi Province. All his such measures not only provided a guarantee for local TCM development, but also exerted an important impact on local provincial medical structure of the day.


[Chemical constituents of Pogostemon cablin (Blanco) Benth].
Nine compounds were isolated from the herb of Pogostemon cablin. They were identified as patchouli alcohol (1), pogostone (2), friedelin (3), epifriedelinol (4), pachypodol (5), retusine (6), oleanolic acid (7), beta-sitosterol (8) and daucosterol (9) on the basis of spectral data (1H-NMR, IR, MS and UV). Compound 3, 4, 6, 7, 8 and 9 were obtained from the species for the first time.


A new biochemical engineering approach to the fractional precipitation of proteins.
A biochemical engineering framework for optimizing the design and operation of fractional protein precipitation has been developed. The method utilizes a fractionation diagram to represent the purification of a product protein relative to total contaminating protein. The purification factor for a single or double-cut fractional precipitation is obtained as the gradient of an appropriate operating tie-line. A computer algorithm has been devised to maximize the tie-line gradient for a given yield enabling a plot of optimum purification factor versus yield to be constructed. The recovery of the enzyme alcohol dehydrogenase from clarified bakers homogenate using saturated ammonium sulphate has been examined. Fractionation and purification versus yield diagrams were used to investigate the effects of such process parameters as pH, temperature, and initial total protein concentration on fractionation efficiency. The results are discussed in terms of the underlying solubility and mixing phenomena and the industrial application of fractional precipitation.


Trans-catheter aortic valve implantation: Contemporary practice and the future.
With increasing life expectancy, the epidemic of valvular heart disease, especially aortic stenosis (AS), is becoming more prevalent. Transcatheter aortic valve implantation (TAVI) has emerged as an alternative therapy for patients with significant aortic valve disease. It offers a less invasive procedure in comparison to surgical aortic valve replacement (sAVR) and an attractive substitute from the patient's perspective. The evidence for TAVI in inoperable and high risk surgical patients is now established and in the intermediate risk group has been accumulating rapidly and is looking favourable for TAVI. However, the full 'TAVI story' is still unfolding. Technological advances in devices and delivery systems are evolving with the aim to improve the function and durability of TAVI and to simplify the procedure while enhancing safety. The incidence of vascular injury and pacemaker requirement post TAVI remains an issue and further development in this regard is therefore of utmost importance, particularly as lower risk and potentially younger patients are treated. Moreover, the evidence concerning long-term durability of the TAVI prostheses continues to accumulate. Whilst TAVI is proving to be an invaluable tool for inoperable and high risk patients, more trial evidence is needed before it encompases lower risk populations and moreover, its use as a first line treatment worldwide in most healthcare systems is limited by the costs associated with the prosthesis.


A hollow-fiber reactor design for NMR studies of microbial cells.
A hollow-fiber membrane reactor was designed and constructed to allow perfusion of entrapped, dense Escherichia coli cells with nutrient medium during examination of cell metabolism using nuclear magnetic resonance (NMR) spectroscopy. Phosphorus-31 NMR spectra of the perfused cells included peaks for nucleoside di- and triphosphates, sugar phosphates, and pH-sensitive peaks for inorganic phosphate. The observed intensity of the lumenal inorganic phosphate peak was found to depend on flow rate, ruling out the use of this peak as a concentration reference. Absolute intracellular pH values obtained from NMR measurements were found to be accurate to 0.2 pH units due to uncertainties in intracellular ionic concentrations. Relative pH values, however, were found to be sensitive to cell energetic status. The response of E. coli intracellular pH following a shift to carbon starvation medium was monitored with a resolution of 3 min. Use of a hollow-fiber reactor for cell containment and perfusion during NMR spectroscopy enables metabolic experiments of longer duration and of greater variety than is possible using standard, nonperfused sample tubes.


Hospitalization risk associated with typical and atypical antipsychotic use in community-dwelling elderly patients.
Due to age-related changes in drug disposition and response, elderly patients are more susceptible to the adverse effects of antipsychotic medications than younger adults. However, few studies have examined the impact of typical and atypical antipsychotic use on all-cause hospitalization in the elderly population. This study compared the short-term effects of incident use of typical and atypical antipsychotic agents on the risk for hospitalization in a community-dwelling elderly population. This retrospective data analysis involved a longitudinal cohort of typical and atypical antipsychotic users and was based on data from the 1996-2004 Medical Expenditure Panel Survey. Typical antipsychotic agents included chlorpromazine, fluphenazine, haloperidol, levomepromazine, loxapine, mesoridazine, molindone, perphenazine, promazine, thioridazine, thiothixene, and trifluoperazine. Atypical antipsychotic agents included aripiprazole, clozapine, olanzapine, quetiapine, risperidone, and ziprasidone. Incident cases of antipsychotic use in community-dwelling elderly (aged > or =60 years) persons were selected for the assessment of risk for all-cause hospitalization within 60 days of exposure to antipsychotics. Bivariate analyses were used to compare baseline characteristics; multivariate logistic regression was used to compare hospitalization risk among users of typicals and atypicals after controlling for age, sex, race, income, insurance coverage, perceived general health, perceived mental health, and other concurrent psychotropic use. The analytical sample consisted of 124 community-dwelling elderly patients (atypicals, 75 patients; typicals, 49). A majority of the elderly study sample were women (63%), white (79%), and of middle/high income (57%). The mean (SD) age of the study sample was 74.37 (8.65) years. There were no significant differences in baseline characteristics between typical and atypical users, with the exception of perceived mental health status. After controlling for other factors, the risk for hospitalization was nearly 4-fold higher with typical antipsychotic use than atypical use (odds ratio, 3.81; 95% CI, 1.12-12.99). In this population of community-dwelling elderly, use of typical agents was associated with an increased risk for hospitalization compared with atypical agents.


Orthodontic post-adjustment pain control with acupuncture.
This study aimed to evaluate the analgesic efficacy of systemic acupuncture therapy on the pain caused after orthodontic adjustments. An initial sample of 30 orthodontic patients with fixed appliances monthly adjusted was selected; however, only 11 participants completed the study. For this reason, final sample comprised these patients' data only. Initially, average pain levels were assessed at different periods by means of an analogue visual scale (VAS) for three months without acupuncture. In the following three months, the volunteers were submitted to systemic acupuncture sessions on Hegu (LI4) and Jiache (St6) points, before orthodontic adjustments were carried out. Results revealed statistically significant reduction in pain level indexes both for men (P = 0.030) and women (P = 0.028) when acupuncture therapy was performed prior to orthodontic adjustment. Patients did not present any side effects. Acupuncture is a safe and effective method in reducing orthodontic post-adjustment pain.


Riparian Collembola (Hexapoda) communities of northern Moldova, Eastern Europe.
Collembola were studied in a well-preserved riverine section of the Prut River in the Republic of Moldova. The study was focused on species diversity and habitat preferences of the particular species at two localities. Riparian habitats of the Prut River near Branişte included open river bank, forest belt and meadow, and the shore of Lake Costeşti-Stânca included meadow, pasture and shrub vegetation. In total 77 collembolan species were recorded, of which Neanura moldavica and Arrhopalites prutensis were endemic to Moldova. Comparative analyses showed a specific community composition at Branişte, with Anurida ellipsoides and Mesaphorura macrochaeta being abundant on the river bank and Hemisotoma thermophila in the meadow. In contrast, the forest plantation at the same locality was similar to the shrub-land in Costeşti, with the common species Mesaphorura critica, M. yosii, Deutonura albella and Isotomiella minor. Hygrophilous species preferred the habitats of the river section in Branişte, with quiet backwaters, to the artificial shoreline of the large lake. Species diversity was relatively high in the natural meadow and forest in Branişte and also in shrub-land on the lake shore. The present study documented relatively high collembolan species diversity at the shoreline and running water sections in the upper catchment area of the Prut River in Moldova that involve naturally valuable inundated habitats of Eastern Europe.


Bronchial measurement with three-dimensional quantitative thin-section CT in patients with cystic fibrosis.
To prospectively compare bronchial measurements obtained with three-dimensional quantitative thin-section computed tomography (CT) with those obtained with thin-section CT scores in the assessment of the severity of pulmonary cystic fibrosis (CF). Ethics committee approval was obtained. Sixteen patients with CF (mean age, 26.6 years; range, 18-42 years) and five healthy volunteers (mean age, 27.4 years; range, 21-44 years) gave written informed consent, underwent multi-detector row CT and a pulmonary function test (PFT), and were divided into three groups: group A, healthy volunteers; group B, patients with mild CF (forced expiratory volume in 1 second [FEV(1)] > 80%); and group C, patients with severe CF (FEV(1) < 80%). Two observers obtained thin-section CT scores with eight scoring systems. Bronchial cross-sectional wall area (WA), lumen area (LA), airway area, and wall thickness (WT) were measured with customized software and were normalized on the basis of subject body surface. Morphologic characteristics, PFT results, thin-section CT scores, and quantitative measurements were compared among the three groups with analysis of variance. Correlations among bronchial measurements, PFT results, and CT scores were calculated with the Spearman correlation coefficient. Thin-section CT scores were different between group C and either group A or group B (P < .05). WA and WT were significantly different among all groups (P < .05). Interscore correlations and correlations between bronchial parameters and scores were high (r > 0.89, P < .0001). Scores, WA, and WT were significantly correlated with PFT obstructive indexes (P < .047). WA and WT assessed with dedicated software on multi-detector row CT images allow evaluation of the severity of pulmonary CF.


Gene expression profile in liver of differing ages of rats after single oral administration of acetaminophen.
In order to verify the influence of the rat age on hepatotoxicity, male Sprague-Dawley rats of 6 (young) and 12 (adult) weeks of age were orally administered acetaminophen (APAP), isoniazid (INH), or carbon tetrachloride (CCl4). Liver samples were obtained in a time-course manner, and changes in gene expression examined by an Affymetrix GeneChip. APAP caused more prominent hepatic injury with respect to pathology and blood biochemistry in adults than in young rats, whereas no obvious age-related differences were observed in INH- or CCl4-treated rats. Comparing gene expression in control rats, CYP3A13 was higher and GSTY2c was lower in adults, suggesting that production of the active metabolite of APAP is higher and its detoxification is lower in adults. The total amount of glutathione and total SH in rat liver was found to be higher in adult rats whereas the extent of its reduction by APAP was larger in adults. A detailed analysis of genes showing age-related differences revealed that some of them were different not in their extent but in their time course, i.e., the stress responses occurred earlier in the young than in the adult, resulting in a difference at 24 hr after dosing. These results suggest that the age-related difference in toxicity would be attributed to a higher expression of CYP3A13, producing the active metabolite of APAP as well as the lower expression of the detoxification enzyme, GSTY2c, in adult rats. Furthermore, these differences affect the time course of APAP toxicity. The present study clearly depicts the advantage of the multi-time, multi-dose protocol employed in our project for analyzing the mechanism of toxicity by gene expression profiling.


Density Functional Theory Assessment of the Environment Polarity Effect on Polyaniline-Water Coupling.
Crystallization water plays an important role in the self-organization of oligomer chains in conducting polyaniline. In order to quantify the interaction between emeraldine salt and such a water, models containing a tetramer in bipolaronic or polaronic form, chloride counterions, and an explicit water molecule are used. Different initial positions of water with respect to the oligomer chain-tangential and vertical-are considered. Various media are simulated by introducing an implicit solvent continuum of decreasing polarity. The DFT-D3/PCM computational approach is employed to examine the behavior of the systems in several aspects-the role of the explicit water position and the effect of the environment polarity on the spatial structure, energetics, charge distribution, and the frontier molecular orbital energies. The strength of hydrogen bonding and the patterns of charge redistribution invoked by the water molecule are discussed. The study establishes trend lines in the variation of the molecular characteristics upon change of milieu as a tool for control of the self-assembly process. The results show that chains interact more efficiently with tangentially placed water. The influence of the environment polarity is minor and is mainly expressed in slight shortening of the intermolecular distances and mild decrease of the group charges of the system components with reduction of polarity.


Health-care associated infection in the newly-opened intensive care unit.
Health-care associated infections (HAI) are common complications in ICU patients as an effect of high invasive device utilization rate. The aim of the study was to analyze the epidemiology of ventilator - associated pneumonia (VAP), blood stream infections (BSI), urinary tract infections (UTI) and surgical site infections (SSI) in newly opened medical-surgical intensive care unit. VAP, BSI, UTI and SSI were detected and registered by hospital Infection Control Team according to CDC criteria during first 12 months following opening of new ICU. During 12-month period HAI was diagnosed in 44 out of 168 patients (26%). The ventilator utilization rate was 72%. The incidence density of VAP was 15.5/1000 ventilator days. The central line utilization was 100%. The BSI rate was 5/1000 catheter days. The urinary catheter utilization was 95%. The UTI rate was 1.9/1000 catheterization days. In 8 out of 60 surgical patients SSI occurred (13%). The most common isolates were Gram negative bacteria. The incidence of VAP was higher than the mean values reported for USA, western Europe and similar to the mean value reported for developing countries. BSI and UTI rate was comparable to those given in NNIS and HELICS reports. Effective prevention strategies should be introduced in order to reduce high VAP rates


Effect of interferons on protein synthesis in human lymphocytes: enhanced synthesis of eight specific peptides in T cells and activation-dependent inhibition of overall protein synthesis.
Lymphoblastoid and fibroblast IFN inhibited PHA stimulation of overall protein synthesis in human lymphocytes by ca. 30%. Inhibition occurred within the first 6 hr of PHA treatment and was not progressive. DNA synthesis at 48 hr was inhibited to the same extent. Overall protein synthesis in resting lymphocytes was not detectably inhibited by IFN concentrations up to 1000 U/ml. Thus, inhibition of protein synthesis and subsequent reduction of cell proliferation by IFN require certain early events in mitogen activation. Resting lymphocytes were not unresponsive to IFN treatment, however. Two-dimensional electrophoretic analysis of newly synthesized proteins after IFN treatment showed enhanced synthesis of a specific set of eight peptides (I-peptides), which were shown to be synthesized in T lymphocytes. This enhancement was produced by both IFN-alpha and IFN-beta after 4 to 6 hr of exposure and was identical for all lymphocyte donors studied. After growth stimulation, IFN treatment produced no enhancements of additional peptides, although the original eight I-peptides were enhanced as usual. It is concluded that the biochemical activities of the I-peptides, which remain to be determined, cannot inhibit protein synthesis in resting lymphocytes, but may do so after mitogen activation, when the major physiologic restriction of lymphocyte protein synthesis is released. Alternatively, the I-peptides may be unrelated to regulation of protein synthesis but may be involved in viral protection or enhancement of NK activity.


The esperanza window trap reduces the human biting rate of Simulium ochraceum s.l. in formerly onchocerciasis endemic foci in Southern Mexico.
The Esperanza Window Trap (EWT) baited with CO2 and human sweat compounds is attractive to Simulium ochraceum s.l., the primary vector of Onchocerca volvulus in the historically largest endemic foci in México and Guatemala. The ability of the EWT to locally reduce numbers of questing S. ochraceum s.l. was evaluated in two formerly onchocerciasis endemic communities in Southern México. At each community, two EWTs were placed in or near a school or household and flies were collected sequentially for a total of 10 days. Black fly collections were then carried out for an additional 10 days in the absence of the EWTs. Flies were also collected outside the dwellings to control for variations in the local fly populations. When the EWTs were present, there was a significant reduction in the human biting rate at both the household and school locations at collection sites, with a greater effect observed in the schools. These results indicate that the EWTs not only have potential as a black fly monitoring tool but may be used for reducing personal exposure to fly bites in Mesoamerica.


Effects of ruthenium red intraperitoneally injected to adult rats, on the uptake and release of neurotransmitters from brain synaptosomes.
Effect of intraperitoneal (i.p.) injection of ruthenium red (RuR) to adults rats on catecholamine (CA) uptake and dopamine (DA) release was evaluated in brain synaptosomal fractions. No effect on CA uptake by synaptosomal fractions was seen when RuR was added in vitro. There was no effect of RuR injection i.p. to rats, on CA uptake by their brain synaptosomal fractions. A blocking effect on depolarizing induced released of DA from brain synaptosomal fraction obtained from rats previously injected i.p. with RuR was seen. A similar effect was observed on the basal DA efflux from synaptosomes loaded with (14C)DA obtained from rats i.p. injected with RuR. It is concluded that RuR, a non-liposoluble substance, when injected i.p. reaches the brain parenchyma through areas devoid of blood-brain barrier and interferes with Ca(++)-dependent release of neurotransmitters, inducing cerebral hyperexcitability and convulsions.


The effect of images on item statistics in multiple choice anatomy examinations.
Although multiple choice examinations are often used to test anatomical knowledge, these often forgo the use of images in favor of text-based questions and answers. Because anatomy is reliant on visual resources, examinations using images should be used when appropriate. This study was a retrospective analysis of examination items that were text based compared to the same questions when a reference image was included with the question stem. Item difficulty and discrimination were analyzed for 15 multiple choice items given across two different examinations in two sections of an undergraduate anatomy course. Results showed that there were some differences item difficulty but these were not consistent to either text items or items with reference images. Differences in difficulty were mainly attributable to one group of students performing better overall on the examinations. There were no significant differences for item discrimination for any of the analyzed items. This implies that reference images do not significantly alter the item statistics, however this does not indicate if these images were helpful to the students when answering the questions. Care should be taken by question writers to analyze item statistics when making changes to multiple choice questions, including ones that are included for the perceived benefit of the students. Anat Sci Educ 10: 68-78. © 2016 American Association of Anatomists.


Single umbilical artery. Correlation of clinical status and umbilical cord histology.
Although numerous reports are available concerning infants who have single umbilical artery, there are no studies that correlate the incidence of congenital malformations or prognosis of such infants with the presence or absence of an atrophied artery in their associated umbilical cords. For this reason, we studied 48 infants who have single umbilical artery. No significant differences were detected in congenital malformations or neonatal mortality between infants with atrophy of one of their arteries and infants with arterial aplasia.


Endocrine disrupter bisphenol a induces orphan nuclear receptor Nur77 gene expression and steroidogenesis in mouse testicular Leydig cells.
The orphan nuclear receptor Nur77 (NR4A1) is a member of the nuclear receptor superfamily, which plays an important role in the regulation of LH-mediated steroidogenesis in testicular Leydig cells. The aim of the current study was to investigate the potential role of bisphenol A (BPA) on orphan nuclear receptor Nur77 gene expression and steroidogenesis. Northern blot analysis demonstrated that BPA transiently induced Nur77 mRNA expression, and protein kinase inhibitor H-89 and PD98059 strongly inhibited the induction of BPA-mediated Nur77 gene expression in mouse Leydig tumor cell line, K28. Moreover, BPA increased the activation of mitogen-activated protein kinase. Transient transfection assay demonstrated that BPA increased Nur77 gene promoter activity and Nur77 transactivation, whereas BPA did not significantly affect the interaction of Nur77 with its corepressor. Furthermore, BPA increased progesterone biosynthesis in K28 cells, which was suppressed by overexpression of dominant negative Nur77. Finally, BPA injection to prepubertal mice revealed that the expression of Nur77 mRNA was elevated, and this induction was correlated with increased concentration of testicular T in vivo. Taken together, these results demonstrated that BPA induces Nur77 gene expression and subsequently alters the steroidogenesis in testicular Leydig cells. These observations provide a novel mechanism by which BPA acts as an endocrine disrupting chemical.


Applications of nonlocal means algorithm in low-dose X-ray CT image processing and reconstruction: A review.
Low-dose X-ray computed tomography (LDCT) imaging is highly recommended for use in the clinic because of growing concerns over excessive radiation exposure. However, the CT images reconstructed by the conventional filtered back-projection (FBP) method from low-dose acquisitions may be severely degraded with noise and streak artifacts due to excessive X-ray quantum noise, or with view-aliasing artifacts due to insufficient angular sampling. In 2005, the nonlocal means (NLM) algorithm was introduced as a non-iterative edge-preserving filter to denoise natural images corrupted by additive Gaussian noise, and showed superior performance. It has since been adapted and applied to many other image types and various inverse problems. This paper specifically reviews the applications of the NLM algorithm in LDCT image processing and reconstruction, and explicitly demonstrates its improving effects on the reconstructed CT image quality from low-dose acquisitions. The effectiveness of these applications on LDCT and their relative performance are described in detail.


Involvement of selective epithelial cell death in the formation of feather buds on a bioengineered skin.
Selective cell death by apoptosis plays important roles in organogenesis. Apoptotic cells are observed in the developmental and homeostatic processes of several ectodermal organs, such as hairs, feathers, and mammary glands. In chick feather development, apoptotic events have been observed during feather morphogenesis, but have not been investigated during early feather bud formation. Previously, we have reported a method for generating feather buds on a bioengineered skin from dissociated skin epithelial and mesenchymal cells in three-dimensional culture. During the development of the bioengineered skin, epithelial cavity formation by apoptosis was observed in the epithelial tissue. In this study, we examined the selective epithelial cell death during the bioengineered skin development. Histological analyses suggest that the selective epithelial cell death in the bioengineered skin was induced by caspase-3-related apoptosis. The formation of feather buds of the bioengineered skin was disturbed by the treatment with a pan-caspase inhibitor. The pan-caspase inhibitor treatment suppressed the rearrangement of the epithelial layer and the formation of dermal condensation, which are thought to be essential step to form feather buds. The suppression of the formation of feather buds on the pan-caspase inhibitor-treated skin was partially compensated by the addition of a GSK-3β inhibitor, which activates Wnt/β-catenin signaling. These results suggest that the epithelial cell death is involved in the formation of feather buds of the bioengineered skin. These observations also suggest that caspase activities and Wnt/β-catenin signaling may contribute to the formation of epithelial and mesenchymal components in the bioengineered skin.


Risk evaluation and mitigation strategies: a focus on belatacept.
To review the elements and components of the risk evaluation and mitigation strategies (REMS) for the costimulation blocker belatacept and associated implications for health care providers working with transplant recipients. The MEDLINE and EMBASE databases (January 1990 to March 2012) were searched by using risk evaluation and mitigation strategies, REMS, belatacept, and organ transplant as search terms (individual organs were also searched). Retrieved articles were supplemented with analysis of information obtained from the Federal Register, the Food and Drug Administration, and the manufacturer of belatacept. REMS are risk-management strategies implemented to ensure that a product's benefits outweigh its known safety risks. Although belatacept offers a novel strategy in maintenance immunosuppression and was associated with superior renal function compared with cyclosporine in phase 2 and 3 trials, belatacept is also associated with increased risk of posttransplant lymphoproliferative disorder and central nervous system infections. The Food and Drug Administration required development of a REMS program as part of belatacept's approval process to ensure safe and appropriate use of the medication and optimization of its risk-benefit profile. Elements of the belatacept REMS include a medication guide that must be dispensed with each infusion and a communication plan. In the management of a complex population of patients, it is essential that those who care for transplant recipients, and patients, recognize the implications of potential and known risks of belatacept. The REMS program aims to facilitate careful selection and education of patients and vigilant monitoring.


The effectiveness of transcutaneous tibial nerve stimulation (TTNS) for adults with overactive bladder syndrome: A systematic review.
To evaluate effectiveness of transcutaneous tibial nerve stimulation (TTNS) for treating adults with overactive bladder (OAB) of idiopathic or neurogenic origin, using a systematic review of the literature. Systematic searches of four databases were undertaken between 1980 and 2017. Included studies investigated effects of TTNS on OAB. Study selection, data extraction, quality appraisal was performed by two independent reviewers. Narrative analysis was undertaken where meta-analysis was not possible due to study heterogeneity. Meta-analysis of RCTs was performed using a fixed effects model. Ten RCTs and three prospective cohort studies involving 629 participants were reviewed. Meta-analysis of two trials comparing TTNS with sham showed mean reduction in total ICIQ Urinary Incontinence Short Form (ICIQ-UI SF) associated with TTNS of -3.79 (95% CI -5.82, -1.76; P = 0.0003, I2 = 25%). Narrative review showed TTNS and antimuscarinic treatment were equally effective (four trials), TTNS provided greater benefit for OAB symptoms than behavioral interventions (two trials), tibial nerve, and sacral foramen stimulation were equally effective but combined stimulation was most effective (one trial). Significant improvements in OAB symptoms were reported by 48-93% participants and UI cure rates of 25-45%. No adverse events were reported. Limited evidence is provided that TTNS is an effective, safe intervention for idiopathic OAB in adults and may be of benefit in those with neurogenic OAB. Further studies are essential to confirm these results as well as to determine efficacy and associated costs for specific patient groups, most effective stimulation dosage, duration of effect, and stimulation regimes for longer-term maintenance.


Treatment with acyclovir does not affect orogenital ulcers in Behçet's syndrome: a randomized double-blind trial.
Acyclovir is a potent antiviral drug, of proven efficacy in the treatment of herpes simplex virus infection. Since this virus has been implicated in the aetiology of Behçet's syndrome, the effect of acyclovir on orogenital ulceration in this condition was determined in a randomized, double-blind, placebo-controlled, crossover trial. Eighteen of 22 patients entering the study completed the trial. Treatment with acyclovir failed to alleviate the frequency and severity of orogenital ulceration or other disease features.


Detection of polyomavirus BK and JC in children with kidney diseases and renal transplant recipients.
Reactivation of polyomavirus is a known reason for severe renal dysfunction in adult renal transplant recipients. Testing for polyomavirus DNA in plasma has been described as a sensitive and specific method to discover viral nephropathy in adult patients. We were now interested in polyomavirus status in a pediatric patient setting. Plasma and urine samples were obtained from 80 children including 38 children after renal transplantation (group 1), 7 children with different kidney diseases receiving immunosuppressive treatment (group 2) and 35 children with different kidney diseases not receiving immunosuppressive treatment (group 3). A nested polymerase chain reaction method was used for amplification of polyomavirus DNA fragments. Differentiation between JC and BK virus was done by digestion with restriction endonucleases. Polyomavirus DNA was detected in the urine sample of 19 of 38 (50%) renal transplant recipients (group 1), of 1 of 7 (14%) patients from group 2 and in none of the 35 patients of group 3. Plasma samples from 3 (8%) of group 1 patients and from 1 child each of group 2 (14%) and group 3 (3%) were tested positive for polyomavirus DNA. Urinary polyomavirus excretion seems to be more frequent in pediatric patients with kidney diseases receiving immunosuppressive treatment and after renal transplantation than in children with various kidney diseases without immunosuppressive treatment.


Determination of 14C-labelled ketone bodies by liquid-scintillation counting.
1. A method of assaying (14)C in ketone bodies present in blood by using liquid-scintillation counting is described. 2. d(-)-beta-Hydroxy[(14)C]butyrate is converted quantitatively into [(14)C]acetoacetate by means of a coupled oxidoreduction reaction involving NAD(+), d(-)-beta-hydroxybutyrate dehydrogenase and malic dehydrogenase in the presence of a high concentration of oxaloacetate. 3. [(14)C]Acetoacetate is decarboxylated to acetone and carbon dioxide which are trapped separately in a double-well flask and counted subsequently. 4. The method permits the determination of (14)C activity in the individual ketone bodies and allows the activity in the carboxyl carbon atoms of acetoacetate or of d(-)-beta-hydroxybutyrate to be assayed separately from the activity in the remainder of the molecule. 5. Recoveries of (14)C-labelled ketone bodies added to blood approach 100% with good reproducibility in replicate analyses.


Treatment of hypertension in the elderly: a time for caution?
The authors review recent studies of treatment of hypertension and suggest modified guidelines for the treatment of elderly hypertensive patients. Treatment is recommended for elderly persons with systolic-diastolic hypertension if the systolic pressure is consistently above 160 mm Hg or the diastolic blood pressure is consistently above 100 mm Hg. Treatment of isolated systolic hypertension should not be excessively vigorous, and systolic blood pressure should not be lowered much below 160 mm Hg.


Lack of immunosuppressive effect of low-dose oral methotrexate on lymphocytes in rheumatoid arthritis.
Whether methotrexate (MTX) is effective in rheumatoid arthritis (RA) because of immunosuppressive and/or anti-inflammatory mechanisms of action is controversial. Many lines of investigation point to the latter. We evaluated DNA synthesis in peripheral blood lymphocytes (PBL) from 33 RA patients on oral MTX (7.5-15 mg/wk) and in 30 healthy controls by flow cytometric cell cycle analysis (CCA). DNA synthesis was also evaluated with a thymidilate synthetase activity assay (TSA) (3H-deoxyuridine incorporation) in 12 patients and 21 controls (12 on MTX and NSAID, and 9 healthy subjects). The patients had taken MTX for at least 3 months and were in different stages of clinical activity. There were no significant differences in TSA or in the cell cycle phase distributions (especially the S phase) between treated RA patients and controls. These data suggest that low-dose oral MTX does not inhibit DNA synthesis and therefore does not have an immunosuppressive effect on lymphocytes from patients with RA.


Biochemical diversity among sulfur-dependent, hyperthermophilic microorganisms.
Hyperthermophiles are a recently discovered group of microorganisms that grow at and above 90 degrees C. They currently comprise over 20 different genera, and except for two novel bacteria, all are classified as Archaea. The majority of these organisms are obligately anaerobic heterotrophs that reduce elemental sulfur (S degree) to H2S. The best studied from a biochemical perspective are the archaeon, Pyrococcus furiosus, and the bacterium, Thermotoga maritima, both of which are saccharolytic. P. furiosus is thought to contain a new type of Entner-Doudoroff pathway for the conversion of carbohydrates ultimately to acetate, H2 and CO2. The pathway is independent of nicotinamide nucleotides and involves novel types of ferredoxin-linked oxidoreductases, one of which has tungsten, a rarely used element, as a prosthetic group. The only site of energy conservation is at the level of acetyl CoA, which is the presence of ADP and phosphate is converted to acetate and ATP in a single step. In contrast, T. maritima utilizes a conventional Embden-Meyerhof pathway for sugar oxidation. P. furiosus also utilizes peptides as a sole carbon and energy source. Amino acid oxidation is thought to involve glutamate dehydrogenase together with at least three types of novel ferredoxin-linked oxidoreductases which catalyze the oxidation of 2-ketoglutarate, aryl pyruvates and formaldehyde. One of these enzymes also utilizes tungsten. In P. furiosus, virtually all of the reductant that is generated during the catabolism of both carbohydrates and peptides is channeled to a cytoplasmic hydrogenase. This enzyme is now termed sulhydrogenase, as it reduces both protons to H2 and S degrees (or polysulfide) to H2S. S degrees reduction appears to lead to the conservation of energy in P. furiosus but not in T. maritima, although the mechanism by which this occurs is not known.


Comparative studies of the biological activities of human tumor necrosis factor and its derivatives.
Biological activities of human tumor necrosis factor (TNF) and its derivatives were compared. In cytotoxicity assay with L929 cells, one derivative, designated as TNF(Asn), showed significantly lower activity than any other TNF examined. In binding assay, this derivative was also shown to have lower affinity for TNF receptors on L929 cells, suggesting that the cytotoxic activity of TNFs on L929 cells correlates with their affinity for receptors. We also found that the cytotoxic activity of TNF on A673 cells and its inhibitory effect on lipoprotein lipase were parallel with the cytotoxic activity on L929 cells, but the growth-enhancing activity on FS-4 cells and the cytotoxic activity on endothelial cells were not. It was also shown that TNF(Asn) had lower affinity than any other TNF for receptors on these target cells tested. These results suggested that there might be at least two types of cellular responses to TNF; one might correlate with the receptor-binding affinity of TNFs and the other not.


Two Rab5 Homologs Are Essential for the Development and Pathogenicity of the Rice Blast Fungus Magnaporthe oryzae.
The rice blast fungus, Magnaporthe oryzae, infects many economically important cereal crops, particularly rice. It has emerged as an important model organism for studying the growth, development, and pathogenesis of filamentous fungi. RabGTPases are important molecular switches in regulation of intracellular membrane trafficking in all eukaryotes. MoRab5A and MoRab5B are Rab5 homologs in M. oryzae, but their functions in the fungal development and pathogenicity are unknown. In this study, we have employed a genetic approach and demonstrated that both MoRab5A and MoRab5B are crucial for vegetative growth and development, conidiogenesis, melanin synthesis, vacuole fusion, endocytosis, sexual reproduction, and plant pathogenesis in M. oryzae. Moreover, both MoRab5A and MoRab5B show similar localization in hyphae and conidia. To further investigate possible functional redundancy between MoRab5A and MoRab5B, we overexpressed MoRAB5A and MoRAB5B, respectively, in MoRab5B:RNAi and MoRab5A:RNAi strains, but neither could rescue each other's defects caused by the RNAi. Taken together, we conclude that both MoRab5A and MoRab5B are necessary for the development and pathogenesis of the rice blast fungus, while they may function independently.


THE IMPACT OF PROFESSIONAL STRESS ON THE MENTAL HEALTH OF LAW ENFORCEMENT OFFICERS.
The aim: To investigate the impact of professional stress on the mental health of law enforcement officers. Materials and methods: The research involved two groups of law enforcement officers (aged 25-40): the first group with up to 7 years of service activities (n = 52), the second group - 8-15 years (n = 48). Research methods: bibliosemantic, questionnaire, testing, method of system analysis and generalization, mathematical and statistical. Results: It was found that law enforcement officers of both groups identified indicators of a significant professional workload, lack of adequate rest, and the nature of communication with participants in professional relationships among the factors of professional stress. It is proved that the mental health of law enforcement officers deteriorates under the influence of professional stress with the experience of their professional activities. Law enforcement officers with 8-15 years of service experience have significantly (p < 0.05-0.001) worse indicators of anxiety, aggressiveness, motivation, personal detachment, impulsive¬ness, vigor, and work efficiency. Conclusions: It was established that the professional stress of law enforcement officers is the result of an imbalance between the internal resources of the individual and the requirements of the external environment, embodied in the peculiarities of law enforcement activities. Law enforcement officers are char¬acterized by the presence of professional stress of average and high severity, which increases with the experience of their service activities. A set of measures was proposed to provide psychological assistance to law enforcement officers to effectively counteract professional stress and improve their mental health.


Patterns of recurrent hepatitis C after liver transplantation in a recent cohort of patients.
Clinicopathologic trends of recurrent hepatitis C after liver transplantation (LT) in hepatitis C (HCV) patients seem to have changed in recent years. Our aims were to define the current post-LT patterns of HCV recurrence and identify features of diagnostic and/or prognostic significance. Detailed analysis was performed on 92 HCV patients who underwent LT from June 1999 to December 2003 and survived early post-LT period. The study patients were grouped, as follows: no histologic recurrence (n = 31), "typical" recurrent HCV (n = 52), and post-LT autoimmune-like hepatitis ("AIH-like") (n = 9). The typical and AIH-like groups had mostly common features with post-LT progressive fibrosis (stage > or =2) more frequent in the latter. Based on post-LT progressive fibrosis (stage > or =2), the 2 post-LT hepatitis categories were regrouped as progressive (n = 24) and nonprogressive (n = 37). High viral counts, HCV genotype 1, and native liver inflammation grade 2 or higher with plasmacytic periseptitis were more frequent in progressive cases than nonprogressive or nonrecurrent cases. Sex mismatch of male recipient and female donor was more common in nonrecurrent group. Overall, death rate was comparable in all groups; however, post-LT HCV-related deaths were more common in progressive cases. In conclusion (1) two thirds (66.2%) of HCV patients developed histologic hepatitis after LT with either typical or AIH-like features; (2) progressive fibrosis was seen in 39.3% of patients with post-LT hepatitis and 26% of the entire study group and was more frequent in AIH-like cases; (3) inflammation grade 2 or higher with plasmacytic periseptitis in native livers may be a predictor of post-LT progressive fibrosis; and (4) male recipient/female donor combination was more common in nonrecurrent cases.


Lipidomic Profiling for Serum Biomarkers in Mice Exposed to Ionizing Radiation.
Radiation biodosimeters are required urgently for fast and accurate evaluation of absorbed dose for irradiated individuals. Lipidomics has appeared as a credible technique for identification and quantification of lipid for researching biomarker of diseases. We performed a lipidomic profile on mice serum at time points of 6, 24, and 72 hours after 0, 2, 5.5, 7, and 8 Gy irradiation to select radiation-responsive lipids and conducted Kyoto Encyclopedia of Genes and Genome pathway enrichment analysis to recognize the pathways and network changes. Then, Pearson correlation analysis was performed to evaluate the feasibility of radiation-responsive lipids to estimate radiation dose. Seven radiation-responsive lipids including PC (18:2/18:2), PC (18:0/18:2), Lyso PC 18:1, PC (18:0/20:4), SM (D18:0/24:1), PC (16:0/18:1), and Lyso PC 18:2 were identified in which glycerophospholipid metabolism presented as the most significant pathway, and they all presented good linear correlation with the irradiated dose. This study identified 7 radiation-responsive lipids in mice serum and certificate their feasibility of dose estimation as biodosimeters.


Evaluation of an enzyme immunoassay kit for estrogen receptor measurement--report II.
We present evidence to show that monoclonal antibodies to estrogen receptors (ER) in solid phase recognize the secondary estrogen binding sites with moderate to low affinity for estradiol (E2). An excellent quantitative agreement was found in five cytosols between the ER values obtained by the enzyme immunoassay (ER-EIA) and the amount of secondary estrogen binding sites measured by the assay involving dextran-coated charcoal (Clin Chem 1986;32:1496). The immunoreactive protein recognized by the antibody-coated beads, when allowed to react with ER(+) cytosols, is shown to bind [3H]estradiol only when the ligand concentration exceeds 8 nmol/L. Further biochemical and functional characterization of the immunoreactive protein is required to establish similarities/dissimilarities between this protein, high-affinity Type I ER sites, and the secondary sites such as Type II sites.


A smartphone readout system for gold nanoparticle-based lateral flow assays: application to monitoring of digoxigenin.
For modern approaches in precision medicine, fast and easy-to-use point-of-care diagnostics (POCs) are essential. Digoxin was chosen as an example of a drug requiring close monitoring. Digoxin is a cardiac glycoside used for the treatment of tachycardia with a narrow therapeutic window of 0.5-2.0 ng·mL-1, and toxic effects are common for concentrations above 2.5 ng·mL-1. For monitoring of blood concentration levels and treatment of intoxication, highly selective antibodies for digoxin and its hapten, digoxigenin, are available. A smartphone readout system is described for measuring digoxigenin in human serum using a common gold nanoparticle lateral flow assay (LFA). The R-package GNSplex, which also includes a Shiny app for quantitative test interpretation based on linear models, is used for image analysis. Images of lateral flow strips were taken with an iPhone camera and a simple darkbox made from black cardboard. Sensitivity and accuracy of the quantitative smartphone system as well as analytical parameters such as limit of detection (LOD) were determined and compared to data obtained with a high resolution BioImager. The data show that the smartphone based digoxin assay yields reliable quantitative results within the clinically relevant concentration range. Graphical abstract For therapeutic drug monitoring and point of care diagnostics we introduce the open source R-package GNSplex for smartphone readout and interpretation of lateral flow assays. The cardiac glycoside dixogin was used as target for this quantitative smartphone reader.


Phenotype and Function of Myeloid-Derived Suppressor Cells Induced by Porphyromonas gingivalis Infection.
Porphyromonas gingivalis, a major etiologic agent of periodontitis, has been reported to induce the expansion of myeloid-derived suppressor cells (MDSC); however, little is known regarding the subpopulations of MDSC expanded by P. gingivalis infection. Flow cytometry was used to evaluate bone marrow and spleen cells from mice infected with P. gingivalis and controls for surface expression of CD11b, Ly6G, and Ly6C. To characterize the phenotype of MDSC subpopulations induced by infection, cells were sorted based on the differential expression of Ly6G and Ly6C. Moreover, since MDSC are suppressors of T cell immune activity, we determined the effect of the induced subpopulations of MDSC on the proliferative response of OVA-specific CD4+ T cells. Lastly, the plasticity of MDSC to differentiate into osteoclasts was assessed by staining for tartrate-resistant acid phosphatase activity. P. gingivalis infection induced the expansion of three subpopulations of MDSC (Ly6G++ Ly6C+, Ly6G+ Ly6C++, and Ly6G+ Ly6C+); however, only CD11b+ Ly6G+ Ly6C++-expressing cells exerted a significant suppressive effect on T cell proliferation. Inhibition of proliferative responses required T cell-MDSC contact and was mediated by inducible nitric oxide synthase and cationic amino acid transporter 2 via gamma interferon. Furthermore, only the CD11b+ Ly6G+ Ly6C++ subpopulation of MDSC induced by P. gingivalis infection was able to differentiate into osteoclasts. Thus, the inflammatory response induced by P. gingivalis infection promotes the expansion of immune-suppressive cells and consequently the development of regulatory inhibitors that curtail the host response. Moreover, monocytic MDSC have the plasticity to differentiate into OC, thus perhaps contributing to the OC pool in states of periodontal disease.


Cryogenic high-resolution X-ray spectrometers for SR-XRF and microanalysis.
Experimental results are presented obtained with a cryogenically cooled high-resolution X-ray spectrometer based on a 141 x 141 micro m Nb-Al-Al(2)O(3)-Al-Nb superconducting tunnel junction (STJ) detector in an SR-XRF demonstration experiment. STJ detectors can operate at count rates approaching those of semiconductor detectors while still providing a significantly better energy resolution for soft X-rays. By measuring fluorescence X-rays from samples containing transition metals and low-Z elements, an FWHM energy resolution of 6-15 eV for X-rays in the energy range 180-1100 eV has been obtained. The results show that, in the near future, STJ detectors may prove very useful in XRF and microanalysis applications.


Comparative analysis of coronavirus genomic RNA structure reveals conservation in SARS-like coronaviruses.
Coronaviruses, including SARS-CoV-2 the etiological agent of COVID-19 disease, have caused multiple epidemic and pandemic outbreaks in the past 20 years1-3. With no vaccines, and only recently developed antiviral therapeutics, we are ill equipped to handle coronavirus outbreaks4. A better understanding of the molecular mechanisms that regulate coronavirus replication and pathogenesis is needed to guide the development of new antiviral therapeutics and vaccines. RNA secondary structures play critical roles in multiple aspects of coronavirus replication, but the extent and conservation of RNA secondary structure across coronavirus genomes is unknown5. Here, we define highly structured RNA regions throughout the MERS-CoV, SARS-CoV, and SARS-CoV-2 genomes. We find that highly stable RNA structures are pervasive throughout coronavirus genomes, and are conserved between the SARS-like CoV. Our data suggests that selective pressure helps preserve RNA secondary structure in coronavirus genomes, suggesting that these structures may play important roles in virus replication and pathogenesis. Thus, disruption of conserved RNA secondary structures could be a novel strategy for the generation of attenuated SARS-CoV-2 vaccines for use against the current COVID-19 pandemic.


A Deterministic Model for Understanding Nonlinear Viral Dynamics in Oysters.
Contamination of oysters with a variety of viruses is one key pathway to trigger outbreaks of massive oyster mortality as well as human illnesses, including gastroenteritis and hepatitis. Much effort has gone into examining the fate of viruses in contaminated oysters, yet the current state of knowledge of nonlinear virus-oyster interactions is not comprehensive because most studies have focused on a limited number of processes under a narrow range of experimental conditions. A framework is needed for describing the complex nonlinear virus-oyster interactions. Here, we introduce a mathematical model that includes key processes for viral dynamics in oysters, such as oyster filtration, viral replication, the antiviral immune response, apoptosis, autophagy, and selective accumulation. We evaluate the model performance for two groups of viruses, those that replicate in oysters (e.g., ostreid herpesvirus) and those that do not (e.g., norovirus), and show that this model simulates well the viral dynamics in oysters for both groups. The model analytically explains experimental findings and predicts how changes in different physiological processes and environmental conditions nonlinearly affect in-host viral dynamics, for example, that oysters at higher temperatures may be more resistant to infection by ostreid herpesvirus. It also provides new insight into food treatment for controlling outbreaks, for example, that depuration for reducing norovirus levels is more effective in environments where oyster filtration rates are higher. This study provides the foundation of a modeling framework to guide future experiments and numerical modeling for better prediction and management of outbreaks. IMPORTANCE The fate of viruses in contaminated oysters has received a significant amount of attention in the fields of oyster aquaculture, food quality control, and public health. However, intensive studies through laboratory experiments and in situ observations are often conducted under a narrow range of experimental conditions and for a specific purpose in their respective fields. Given the complex interactions of various processes and nonlinear viral responses to changes in physiological and environmental conditions, a theoretical framework fully describing the viral dynamics in oysters is warranted to guide future studies from a top-down design. Here, we developed a process-based, in-host modeling framework that builds a bridge for better communications between different disciplines studying virus-oyster interactions.


Chemokine Receptor Ccr7 Restricts Fatal West Nile Virus Encephalitis.
West Nile virus (WNV) is a mosquito-transmitted flavivirus that can cause debilitating encephalitis. To delineate the mechanisms behind this pathology, we studied Ccr7-deficient mice, which afforded us the capacity to study infection in mice with disrupted peripheral cellular trafficking events. The loss of Ccr7 resulted in an immediate pan-leukocytosis that remained elevated throughout the infection. This leukocytosis resulted in a significant enhancement of leukocyte accumulation within the central nervous system (CNS). Despite an excess of virus-specific T cells in the CNS, Ccr7-deficient mice had significantly higher CNS viral loads and mortality rates than wild-type animals. Mechanistically, the elevated trafficking of infected myeloid cells into the brain in Ccr7-deficient mice resulted in increased levels of WNV in the CNS, thereby effectively contributing to neuroinflammation and lowering viral clearance. Combined, our experiments suggest that during WNV infection, Ccr7 is a gatekeeper for nonspecific viral transference to the brain.IMPORTANCE In this study, we show that Ccr7 is required for the sufficient migration of dendritic cells and T cells into the draining lymph node immediately following infection and for the restriction of leukocyte migration into the brain. Further, the severe loss of dendritic cells in the draining lymph node had no impact on viral replication in this organ, suggesting that WNV may migrate from the skin into the lymph node through another mechanism. Most importantly, we found that the loss of Ccr7 results in a significant leukocytosis, leading to hypercellularity within the CNS, where monocytes/macrophages contribute to CNS viremia, neuroinflammation, and increased mortality. Together, our data point to Ccr7 as a critical host defense restriction factor limiting neuroinflammation during acute viral infection.


Assumptions about Decision-Making Capacity and Aphasia: Ethical Implications and Impact.
This article explores the issue of aphasia and decision-making within the context of clinical ethics and patient rights. The cases described illustrate the danger of making assumptions about the inherent competence of people with aphasia and the life-altering consequences if no attempt is made to "accommodate" or support communication when competence may be masked by aphasia. Speech-language pathologists have a moral obligation and a key role to play in providing communication support that may serve to reveal a person's intact capacity to make specific decisions, as well as in supporting the steps involved in the decision-making process. This role also extends to providing guidance, education, and training for others involved in evaluating the decision-making capacity of people with aphasia. Communication support strategies useful at each stage of the decision-making process are detailed.