A fast-and-robust profiler for improving polymerase chain reaction diagnostics.
Polymerase chain reaction (PCR) is an in vitro technology in molecular genetics that progressively amplifies minimal copies of short DNA sequences in a fast and inexpensive manner. However, PCR performance is sensitive to suboptimal processing conditions. Compromised PCR conditions lead to artifacts and bias that downgrade the discriminatory power and reproducibility of the results. Promising attempts to resolve the PCR performance optimization issue have been guided by quality improvement tactics adopted in the past for industrial trials. Thus, orthogonal arrays (OAs) have been employed to program quick-and-easy structured experiments. Profiling of influences facilitates the quantification of effects that may counteract the detectability of amplified DNA fragments. Nevertheless, the attractive feature of reducing greatly the amount of work and expenditures by planning trials with saturated-unreplicated OA schemes is known to be relinquished in the subsequent analysis phase. This is because of an inherent incompatibility of ordinary multi-factorial comparison techniques to convert small yet dense datasets. Treating unreplicated-saturated data with either the analysis of variance (ANOVA) or regression models destroys the information extraction process. Both of those mentioned approaches are rendered blind to error since the examined effects absorb all available degrees of freedom. Therefore, in lack of approximating an experimental uncertainty, any outcome interpretation is rendered subjective. We propose a profiling method that permits the non-linear maximization of amplicon resolution by eliminating the necessity for direct error estimation. Our approach is distribution-free, calibration-free, simulation-free and sparsity-free with well-known power properties. It is also user-friendly by promoting rudimentary analytics. Testing our method on published amplicon count data, we found that the preponderant effect is the concentration of MgCl2 (p<0.05) followed by the primer content (p<0.1) whilst the effects due to either the content of the deoxynucleotide (dNTP) or DNA remained dormant (p>0.1). Comparison of the proposed method with other stochastic approaches is also discussed. Our technique is expected to have extensive applications in genetics and biotechnology where there is a demand for cheap, expedient, and robust information.


Separate developmental programs for HLA-A and -B cell surface expression during differentiation from embryonic stem cells to lymphocytes, adipocytes and osteoblasts.
A major problem of allogeneic stem cell therapy is immunologically mediated graft rejection. HLA class I A, B, and Cw antigens are crucial factors, but little is known of their respective expression on stem cells and their progenies. We have recently shown that locus-specific expression (HLA-A, but not -B) is seen on some multipotent stem cells, and this raises the question how this is in other stem cells and how it changes during differentiation. In this study, we have used flow cytometry to investigate the cell surface expression of HLA-A and -B on human embryonic stem cells (hESC), human hematopoietic stem cells (hHSC), human mesenchymal stem cells (hMSC) and their fully-differentiated progenies such as lymphocytes, adipocytes and osteoblasts. hESC showed extremely low levels of HLA-A and no -B. In contrast, multipotent hMSC and hHSC generally expressed higher levels of HLA-A and clearly HLA-B though at lower levels. IFNγ induced HLA-A to very high levels on both hESC and hMSC and HLA-B on hMSC. Even on hESC, a low expression of HLA-B was achieved. Differentiation of hMSC to osteoblasts downregulated HLA-A expression (P = 0.017). Interestingly HLA class I on T lymphocytes differed between different compartments. Mature bone marrow CD4(+) and CD8(+) T cells expressed similar HLA-A and -B levels as hHSC, while in the peripheral blood they expressed significantly more HLA-B7 (P = 0.0007 and P = 0.004 for CD4(+) and CD8(+) T cells, respectively). Thus different HLA loci are differentially regulated during differentiation of stem cells.


Trends in use and cost of initial cancer treatment in Ontario: a population-based descriptive study.
Cancer incidence and treatment-related costs are rising in Canada. We estimated health care use and costs in the first year after diagnosis for patients with 7 common types of cancer in Ontario to examine temporal trends in patterns of care and costs. We selected patients aged 19-44 years who had received a diagnosis of melanoma, breast cancer (female only), testicular cancer or thyroid cancer, in addition to patients aged 45 years and older who had received a diagnosis of breast (female only), prostate, lung or colorectal cancer, between 1997 and 2007. Patients were identified from the Ontario Cancer Registry. Using linked administrative databases, we determined use and costs of chemotherapy, radiotherapy, cancer-related surgery, other admissions to hospital and home care. We adjusted all costs to 2009 Canadian dollars. We identified 20 821 patients aged 19-44 years and 178 797 patients aged 45 years and older. The greatest increases in costs during the study period were for melanoma, breast cancer, colorectal cancer, lung cancer and prostate cancer (p < 0.05). For prostate and lung cancers, mean costs increased 50% (from $11 490 and $22 037 to $15 170 and $34 473, respectively). Mean costs doubled for breast (from $15 460 and $12 909 to $35 977 and $29 362 for younger and older patients, respectively) and colorectal cancers (from $24 769 to $43 964), and nearly tripled for melanoma (from $3581 to $8934). Costs related to hospital admissions accounted for the largest portion of total costs. The use of chemotherapy, radiotherapy and home care generally increased for all cancers. The significant increase in mean costs of initial cancer treatment among the patients included in this study was primarily due to more patients receiving adjuvant therapy and home care, and to the increasing expenditures for these services and cancer-related surgeries. Understanding trends in health care use and costs can help policy-makers to take the necessary measures to achieve a more accountable, high-performing health care system.


[The process of deinstitutionalization in German State Mental Hospitals. Critical overview of empirical research studies].
The process of discharging previously long-term hospitalized patients from German state mental hospitals, and of transferring these patients to other forms of supported housing has meanwhile come to its end. Therefore, this paper presents an overview of the scientific evaluation of this process. By using clearly defined research methodological criteria, eight empirical studies from several German federal states are assessed focusing on the meaningfulness of their results. Because of the heterogeneity of the aims regarding contents of this transformation process, of the research methodological approaches, and of the characteristics of the assessed patient groups, only few general results could be identified. Chronically mentally ill patients (mostly suffering from schizophrenic disorders) who had been successfully discharged to the community were younger, and showed shorter periods of illness and hospitalization than control patients. After discharge they might have experienced positive changes of their objective and subjective quality of life and of their perceived needs of care. Referred to several other outcome domains, no homogeneous tendencies concerning clinically relevant improvement or deterioration were found. In particular, changes related to psychopathological symptoms and social competencies were generally small. However, mortality and re-hospitalization rates of de-hospitalized patients were considerably high, and ranged between 3-24 %, and 4-46 %, respectively. The assessed studies could not identify replicated predictors of successful de-hospitalization. Female sex and long-term last hospitalization seemed to be important for stable placement in forms of supported housing requiring a higher level of autonomy. From the generally unclear procedural quality of the German deinstitutionalization process, the authors deduce implications for the scientific evaluation of future projects of restructuring mental health care services. Besides the clear definition of empirically assessable aims which should be based on theoretical considerations, it is of utmost priority to guarantee high performance quality of research methodological standards.


CAF hierarchy driven by pancreatic cancer cell p53-status creates a pro-metastatic and chemoresistant environment via perlecan.
Heterogeneous subtypes of cancer-associated fibroblasts (CAFs) coexist within pancreatic cancer tissues and can both promote and restrain disease progression. Here, we interrogate how cancer cells harboring distinct alterations in p53 manipulate CAFs. We reveal the existence of a p53-driven hierarchy, where cancer cells with a gain-of-function (GOF) mutant p53 educate a dominant population of CAFs that establish a pro-metastatic environment for GOF and null p53 cancer cells alike. We also demonstrate that CAFs educated by null p53 cancer cells may be reprogrammed by either GOF mutant p53 cells or their CAFs. We identify perlecan as a key component of this pro-metastatic environment. Using intravital imaging, we observe that these dominant CAFs delay cancer cell response to chemotherapy. Lastly, we reveal that depleting perlecan in the stroma combined with chemotherapy prolongs mouse survival, supporting it as a potential target for anti-stromal therapies in pancreatic cancer.


Just say 'I don't know': Understanding information stagnation during a highly ambiguous visual search task.
Visual search experiments typically involve participants searching simple displays with two potential response options: 'present' or 'absent'. Here we examined search behavior and decision-making when participants were tasked with searching ambiguous displays whilst also being given a third response option: 'I don't know'. Participants searched for a simple target (the letter 'o') amongst other letters in the displays. We made the target difficult to detect by increasing the degree to which letters overlapped in the displays. The results showed that as overlap increased, participants were more likely to respond 'I don't know', as expected. RT analyses demonstrated that 'I don't know' responses occurred at a later time than 'present' responses (but before 'absent' responses) when the overlap was low. By contrast, when the overlap was high, 'I don't know' responses occurred very rapidly. We discuss the implications of our findings for current models and theories in terms of what we refer to as 'information stagnation' during visual search.


Validation of a Commercial Assay and Decision Support Tool for Routine Paclitaxel Therapeutic Drug Monitoring (TDM).
The value of therapeutic drug monitoring (TDM) for paclitaxel (PTX) was recently demonstrated in the largest TDM trial ever conducted in oncology. The trial demonstrated significant reduction in neuropathy when using TDM. Dose adjustment for PTX was based on time above a threshold concentration (Tc>0.05). Tc>0.05 must be calculated with a pharmacokinetic model and complex nonlinear mixed-effects software. The use of the software and chromatographic methods to measure PTX requires specialized expertise. User-friendly methods to quantitate PTX and calculate Tc>0.05 could simplify the introduction of TDM into routine clinical practice. The immunoassay (MyPaclitaxel) was used to quantitate PTX in samples from the clinical trial; the results were used to calculate Tc>0.05 using a stand-alone computer program with a simple, friendly graphical user interface for nonlinear mixed-effects pharmacokinetic calculations (MyCare Drug Exposure Calculator). The resulting dose recommendations from the calculated Tc>0.05 were compared with those using liquid chromatography-ultraviolet detection and NONMEM to examine the efficacy of the simpler tools for TDM. There was a good agreement between the immunoassay and liquid chromatography-ultraviolet detection: Passing-Bablok regression slope was 1.045 and intercept was -6.00, R was 0.9757, and mean bias was -1.77 ng/mL (-2.07 nmol/L). Dosing recommendations were identical for 70% of the cycles and within 10% for 89% of the samples. All Tc>0.05 values were at the same or adjacent medical decision points. MyPaclitaxel assay and MyCare Drug Exposure Calculator are convenient, user-friendly tools that may be suitable for routine TDM of PTX in clinical care.


Larviciding to prevent malaria transmission.
Larviciding refers to the regular application of chemical or microbial insecticides to water bodies or water containers to kill the aquatic immature forms of the mosquito (the larvae and pupae). To summarize research evidence evaluating whether larviciding with chemical or microbial insecticides prevents malaria transmission. We searched the Cochrane Infectious Diseases Group Specialized Register; the Cochrane Central Register of Controlled Trials (CENTRAL), published in the Cochrane Library; MEDLINE; Embase; CAB Abstracts; LILACS; the World Health Organization International Clinical Trials Registry Platform (WHO ICTRP); ClinicalTrials.gov; and the ISRCTN registry up to 6 June 2019. We included cluster-randomized controlled trials (cRCTs), interrupted time series (ITS), randomized cross-over studies, non-randomized cross-over studies, and controlled before-and-after studies (CBAs) that compared larviciding with no larviciding. We independently assessed trials for eligibility and risk of bias, and extracted data. We assessed the certainty of evidence using the GRADE approach. Four studies (one cRCT, two CBAs, and one non-randomized cross-over design) met the inclusion criteria. All used ground application of larvicides (people hand-delivering larvicides); one evaluated chemical and three evaluated microbial agents. Studies were carried out in The Gambia, Tanzania, Kenya, and Sri Lanka. Three studies were conducted in areas where mosquito aquatic habitats were less extensive (< 1 km²), and one where habitats were more extensive (> 1 km²; a cross-over study from The Gambia).For aquatic habitats of less than 1 km², one cRCT randomized eight villages in Sri Lanka to evaluate chemical larviciding using insect growth regulator; and two CBA studies undertaken in Kenya and Tanzania evaluated microbial larvicides. In the cRCT, larviciding across all villages was associated with lower malaria incidence (rate ratio 0.24, 4649 participants, low-certainty evidence) and parasite prevalence (risk ratio (RR) 0.26, 5897 participants, low-certainty evidence) compared to no larviciding. The two CBA studies reported lower malaria prevalence during the intervention period (parasite prevalence RR 0.79, 95% confidence interval (CI) 0.71 to 0.89; 70,902 participants; low-certainty evidence). The Kenyan study also reported a reduction in the incidence of new malaria cases (RR 0.62, 95% CI 0.38 to 1.01; 720 participants; very low-certainty evidence).For aquatic habitats of more than 1 km², the non-randomized cross-over trial using microbial larvicides did not detect an effect for malaria incidence (RR 1.58, 95% CI 0.94 to 2.65; 4226 participants), or parasite prevalence (RR 1.15, 95% CI 0.41 to 3.20; 3547 participants); both were very low-certainty evidence. The Gambia trial also reported the mean haemoglobin level, and there was no difference across the four comparisons (mean difference -0.13, 95% CI -0.40 to 0.13; 3586 participants).We were unable to summarize or pool entomological outcomes due to unreported and missing data. Most controlled studies on larviciding have been performed with microbial agents. Ground larviciding for non-extensive larval habitats may have an effect on malaria transmission, and we do not know if there is an effect in large-scale aquatic habitats. We found no studies using larviciding application techniques that could cover large aquatic habitats, such as aerial spraying using aircraft.


German Medical Data Sciences in Studies in Health Technology and Informatics - Reflections on the Fifth Volume.
Since 2017, the German Society for Medical Informatics, Biometry and Epidemiology e.V. (GMDS) offers the submission of full papers to the annual meetings, optional in Studies in Health Technologies and Informatics (Stud HIT) or in GMS Medical Informatics, Biometrics, and Epidemiology (MIBE). GMDS' aim is to increase the attractiveness of the conference and paper submission process in particular for young scientists and to increase the visibility of the conference. A standardized peer review process was established. Since 2017, a 25-35% of the contributions have been submitted as full papers. A total of 177 papers were published in Stud HTI. With an unofficial journal impact factor of 1.088 (2019) and 0.540 (2020), the papers were cited with a frequency similarly to national medical journals or full paper contributions of International medical informatics conferences.


Expression levels of SOX2, KLF4 and brachyury transcription factors are associated with metastasis and poor prognosis in oral squamous cell carcinoma.
The prognosis of oral squamous cell carcinoma (OSCC) patients is affected by tumor recurrence and metastasis, and cancer stem cells are hypothesized to be involved in these processes. Thus, the aim of the present study was to determine whether the expression levels of five stem cell-related transcription factors, sex determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (Oct4), avian myelocytomatosis viral oncogene homolog (c-Myc), Krüppel-like factor 4 (KLF4) and brachyury, are associated with metastasis and survival in OSCC. Immunohistochemistry was performed to analyze the expression of these proteins in biopsy specimens obtained from 108 OSCC patients. The results revealed that the expression of SOX2, Oct4, KLF4 and brachyury were significantly associated with lymph node metastasis (P=0.002, P=0.031, P=0.003 and P=0.007, respectively). In addition, the expression of KLF4 and brachyury were significantly associated with distant metastasis (P=0.014 and P=0.012, respectively). Furthermore, multivariate analysis revealed that SOX2 and KLF4 are predictive factors for lymph node metastasis [odds ratios (ORs), 4.526 and 4.851, respectively], and KLF4 is also a predictive factor for distant metastasis (OR, 9.607). In addition, OSCC patients with low co-expression of SOX2, KLF4 and brachyury exhibited a significantly lower disease-specific survival rate (78.6 vs. 100%; P=0.025; χ2=5.033) and disease-free survival rate (60.7 vs. 90.9%; P=0.015; χ2=5.897) when compared with OSCC patients with high co-expression of these factors. The results indicate that SOX2, KLF4 and brachyury serve important roles in tumor progression, and these transcription factors may thus represent clinically useful prognostic markers for OSCC.


[Morphological characteristic of erythrocytes in experimental hypervitaminosis A].
This investigation was aimed at the analysis of the shape and morpho-densitometric parameters of the erythrocytes in rats with experimental hypervitaminosis A. Male Wistar rats received 0.64 mg/g (1167 IU/g) of retinol palmitate (RP) in oil solution orally for 11 consecutive days. Rats fed oil alone and intact animals were used as control groups. At days 5 and 6 of the experiment, the first manifestations of hypervitaminosis A were observed (body mass loss, localized erythema and hemorrhages). In contrast to control groups, in rats with hypervitaminosis A, the area of erythrocyte cytoplasm decreased gradually in response to RP administration. Discocyte/spherocyte/stomatocyte ratio also changed dynamically: the proportion of discocytes progressively decreased, while the amount of spherocytes and stomatocytes increased. These results show that excess of the vitamin A alters the erythrocyte membrane structure. Integral optical density of erythrocyte cytoplasm in RP-treated rats as well as in oil-fed rats was lower than in intact animals. This may be an indirect evidence of the fall in erythrocyte hemoglobin content. The changes observed in erythrocytes of RP-treated rats may serve as an additional criterion for evaluation of hypervitaminosis A severity.


Oligocene mammals from Ethiopia and faunal exchange between Afro-Arabia and Eurasia.
Afro-Arabian mammalian communities underwent a marked transition near the Oligocene/Miocene boundary at approximately 24 million years (Myr) ago. Although it is well documented that the endemic paenungulate taxa were replaced by migrants from the Northern Hemisphere, the timing and evolutionary dynamics of this transition have long been a mystery because faunas from about 32 to 24 Myr ago are largely unknown. Here we report a late Oligocene fossil assemblage from Ethiopia, which constrains the migration to postdate 27 Myr ago, and yields new insight into the indigenous faunal dynamics that preceded this event. The fauna is composed of large paenungulate herbivores and reveals not only which earlier taxa persisted into the late Oligocene epoch but also demonstrates that one group, the Proboscidea, underwent a marked diversification. When Eurasian immigrants entered Afro-Arabia, a pattern of winners and losers among the endemics emerged: less diverse taxa such as arsinoitheres became extinct, moderately species-rich groups such as hyracoids continued into the Miocene with reduced diversity, whereas the proboscideans successfully carried their adaptive radiation out of Afro-Arabia and across the world.


Relative adrenal insufficiency amongst hospitalized mild to moderate acute ischemic stroke patients.
Acute ischemic stroke is a stressful condition where there is marked increase in the production of cortisol. In the past, adrenal insufficiency in critically ill patients had been shown to be associated with significant morbidity and mortality. To date, there have been no studies performed to determine its prevalence among patients with acute ischemic stroke. The aim of this study was to determine the prevalence of relative adrenal insufficiency in acute ischemic stroke by utilizing low dose (LD) and standard dose (SD) synacthen tests and to correlate it with inpatient hospital morbidity and mortality. Fifty-eight patients who fulfilled the diagnosis of acute ischemic stroke within 72 hours from the onset of a stroke were subjected to LD (1µg) synacthen test (LDST) and two hours later to SD (250 µg) synacthen test (SDST). Based on an increment of less than 250 nmol/L after LDST, 38 (65.5%) patients had relative adrenal insufficiency. However, using similar criteria with the SDST, only 18 (31.0%) patients had relative adrenal insufficiency. Three patients died during the study period and they had a tendency to have high baseline cortisol levels. The diagnosis of relative adrenal insufficiency in general was not associated with any other significant clinical outcomes. This is the first study demonstrating the prevalence of relative adrenal insufficiency amongst acute ischemic stroke patients. Utilizing the LDST, relative adrenal insufficiency was found more sensitive in detecting relative adrenal insufficiency in patients with acute ischemic stroke as compared to SDST.


Xenobiotic-induced transcriptional regulation of xenobiotic metabolizing enzymes of the cytochrome P450 superfamily in human extrahepatic tissues.
Numerous members of the cytochrome P450 (CYP) superfamily are induced after exposure to a variety of xenobiotics in human liver. We have gained considerable mechanistic insights into these processes in hepatocytes and multiple ligand-activated transcription factors have been identified over the past two decades. Families CYP1, CYP2 and CYP3 involved in xenobiotic metabolism are also expressed in a range of extrahepatic tissues (e.g. intestine, brain, kidney, placenta, lung, adrenal gland, pancreas, skin, mammary gland, uterus, ovary, testes and prostate). Since the expression of the majority of the isoforms appears to be very low in the extrahepatic tissues in comparison with predominant expression in adult liver, the role of the enzymes in overall biotransformation and total body clearance is minor. However, basal expression and up-regulation of extrahepatic CYP enzymes can significantly affect local disposition of xenobiotics or endogenous compounds in peripheral tissues and thus modify their pharmacological/toxicological effects or affect absorption of xenobiotics into systemic circulation. The goal of this review is to critically examine our current understanding of molecular mechanisms involved in induction of xenobiotic metabolizing CYP genes of human families CYP1, CYP2 and CYP3 by exogenous chemicals in extrahepatic tissues. We concentrate on organs such as the intestine, kidney, lung, placenta and skin, which are involved in drug distribution and clearance or are in direct contact with environmental xenobiotics. We also discuss single nucleotide polymorphisms (SNPs) of key CYPs, which at the level of transcription affect expression of the genes in the extrahepatic tissues or are associated with some pathophysiological stages or disorders in the organs.


Conjugated estrogens for the management of bleeding associated with renal failure.
Bleeding is a major complication of uremia. Both cryoprecipitate and desmopressin effectively shorten the prolonged bleeding time and favorably influence clinical bleeding, but the former carries the risk of transmitting blood-borne infectious diseases, and both cryoprecipitate and desmopressin have a short duration of action. Preliminary evidence has suggested that estrogens may be useful, and we therefore performed a randomized, double-blind, crossover trial comparing the effect of conjugated estrogens with that of placebo on hemorrhagic tendencies and the bleeding time in six patients with uremia who were on maintenance hemodialysis. Five daily infusions of placebo or conjugated estrogens were administered at the beginning of one-month trial periods. Estrogen shortened the bleeding time in all six patients. The effect was detectable six hours after the first infusion, reached its maximum in all patients between days 5 and 7, and lasted for 14 days. By day 16 after the last infusion, the bleeding time had returned to base line in four of the six patients. No side effects were noted during or after estrogen infusion. Estrogens did not influence the circulating level of von Willebrand factor or change its multimeric structure. Moreover, the defective platelet aggregation and thromboxane formation observed in the patients were not corrected by estrogens. We conclude that conjugated estrogens are an adequate alternative to cryoprecipitate or desmopressin for the treatment of bleeding associated with renal failure, especially when a longer duration of action is needed and immediate onset of the effect is not essential. The mechanism of action of estrogens remains to be clarified.


Forcing term in single-phase and Shan-Chen-type multiphase lattice Boltzmann models.
Numerous schemes have been proposed to incorporate a bulk forcing term into the lattice Boltzmann equation. In this paper we present a simple and straightforward comparative analysis of five popular schemes [Shan and Chen, Phys. Rev. E 47, 1815 (1993); Phys Rev Lett. 81, 1618 (1998); He et al., Phys. Rev. E 57, R13 (1998); Guo et al., Phys. Rev. E 65, 046308 (2002); Kupershtokh et al., Comput. Math. Appl. 58, 965 (2009)] in which their differences and similarities are identified. From the analysis we classify the schemes into two groups; the behaviors of the schemes in each group are proven to be identical up to second order. Numerical test simulating the two-dimensional unsteady Taylor-Green vortex flow problem demonstrate that all five schemes are of comparable accuracy for single-phase flow. However, for two-phase flow the situation is different, which is demonstrated by incorporating these schemes into different Shan-Chen-type multiphase models. The forcing scheme in the original Shan-Chen (SC) multiphase model turns out to be inaccurate in terms of the resulting surface tension for different density ratios and relaxation times. In the numerical tests, a typical equation of state and interparticle interactions including next-nearest neighbors were incorporated into the SC model. Our results confirm that the surface-tension values obtained from the original SC lattice Boltzmann method (LBM) simulation depend on the value of the relaxation time τ. For τ<0.7Δt, the surface tension agree well with the analytical solutions. However, when τ>0.7Δt, the surface tension turns out to be systematically larger than the analytical one, exceeding it by more than a factor of 2 for τ=2Δt. In contrast, with the application of the scheme proposed by He et al., the SC LBM produces very accurate surface tensions independent of the value of τ. We also found that the densities of the coexisting liquid and gas can be adjusted to match those at thermodynamic equilibrium if the particle interaction term includes next-nearest-neighbor contributions. The obtained results will be useful for further studies of two-phase flow with high density ratios using the SC LBM approach.


Subchronic and chronic toxicity of the new antibiotic cefpirome in rats.
Subchronic and chronic toxicity studies in rats were performed with 3-[(2,3-Cyclopenteno-1-pyridinium)-methyl]- 7-[2-syn-methoximino-2-(2-aminothiazol-4-yl)-acetamido] -ceph-3-em-4- carboxylate (cefpirome, HR 810), a new cephalosporin derivative. In subchronic (14 day) studies cefpirome was intravenously administered in dose levels up to 1500 mg/kg/d with good kidney tolerance. Signs of renal functional impairment were observed (800 and 1500 mg/kg) but histologically no morphological changes could be detected. The chronic intraperitoneal administration (90 day) of cefpirome at dose levels of 400 or 1600 mg/kg/d resulted in some reversible changes in hematology (slight anemia), serum-chemistry parameters (liver), urinalysis (proteinuria), and histopathology (increased numbers and enlargement of lysosomes in proximal tubular epithelia of the kidneys and pigment deposits in follicle epithelia of the thyroids), predominantly in high-dose animals. The "no effect level" is considered to be 100 mg/kg/d in this study.


Evaluation of the analgesic effect of intracameral lidocaine hydrochloride injection on intraoperative and postoperative pain in healthy dogs undergoing phacoemulsification.
To evaluate the intraoperative and postoperative analgesic effects of intracameral lidocaine hydrochloride injection in dogs undergoing phacoemulsification. 12 healthy Beagles with healthy eyes. Dogs were randomly assigned to receive 1 of 2 intracameral injections: 2% lidocaine hydrochloride solution (0.3 mL) or an equivalent amount of balanced salt solution (BSS). All dogs were treated with acepromazine (0.05 mg/kg, i.v.) and cefazolin (30 mg/kg, i.v.), and tropicamide drops were topically applied to the eyes. Anesthesia was induced with propofol and maintained with isoflurane. The initial end-tidal isoflurane concentration was maintained at 1.2%. Heart rate, respiratory rate, arterial blood pressure, esophageal temperature, inspired and end-tidal isoflurane concentrations, and oxygen saturation were recorded every 5 minutes. The allocated agent was injected intracamerally after aspiration of the same volume of aqueous humor. Ten minutes after injection, phacoemulsification was performed. After surgery began, the isoflurane concentration was adjusted according to heart rate and mean arterial blood pressure. Pain scores were recorded before surgery and at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 8, 16, and 24 hours after extubation. Isoflurane requirements were significantly higher in the BSS group than in the lidocaine group. Mean +/- SD time to administration of supplementary analgesia was significantly shorter in the BSS group (1.4 +/- 1.2 hours) than in the lidocaine group (4.9 +/- 1.2 hours). Intracameral lidocaine injection had significant analgesic effects in dogs undergoing cataract surgery. Results of this study suggest the value of intracameral lidocaine injection as an analgesic for intraocular surgery in dogs.


Electroacupuncture promotes synaptic plasticity in rats with chronic inflammatory pain-related depression by upregulating BDNF/TrkB/CREB signaling pathway.
Chronic inflammatory pain (CIP) frequently coincides with depression among patients. The onset and development of pain and depression are associated with altered neural synaptic plasticity. Electroacupuncture (EA) can effectively relieve CIP and depression. However, the underlying mechanisms have not been fully illustrated. To explore whether EA can relieve CIP and depression by regulating hippocampal synaptic plasticity, and the present study offers foundational evidence for the efficacy of EA in treating CIP-related depression (CIPD). Rats were divided into four groups: 0.9% normal saline group, complete Freund's adjuvant (CFA) group, CFA + duloxetine group, and CFA + EA group. Pain hypersensitivity was detected by mechanical withdrawal threshold and thermal paw withdrawal latency, and the depression level was gauged using the open field test, the sucrose preference test, and the forced swimming test. The morphology of the hippocampal neurons was observed using Nissl staining. The protein expression levels of synuclein (Syn), postsynaptic density protein-95 (PSD-95), brain-derived neurotrophic factors (BDNFs), tyrosine-protein kinase B (TrKB), p-TrkB, cAMP response element binding protein (CREB), and p-CREB were measured by western blotting and immunofluorescence staining. BDNF and TrkB mRNA expression were detected using quantitative real-time polymerase chain reaction (PCR) (qRT-PCR). The content of 5-hydroxytryptamine (5-HT) and γ-aminobutyric acid (GABA) was detected using enzyme-linked immunosorbent assay, and the glutamic acid (Glu) content was determined using the ultraviolet colorimetry method. The hippocampal neuron ultrastructure was observed using transmission electron microscopy. EA could alleviate CIP and related depressive behaviors as well as protect the hippocampal neuronal structure from damage and regulate 5-HT/GABA/Glu levels in the hippocampus. Additionally, EA could significantly increase the expression of synapse-associated proteins such as PSD-95 and Syn by activating the BDNF/TrKB/CREB signaling pathway. EA improves pain and depressive behaviors in CIPD rats, and the mechanism may be related to synaptic plasticity mediated by the BDNF/TrKB/CREB signaling pathway.


Nongated Cardiac Computed Tomographic Angiograms for Detection of Embolic Sources in Acute Ischemic Stroke.
We assessed the feasibility of obtaining diagnostic quality images of the heart and thoracic aorta by extending the z axis coverage of a non-ECG-gated computed tomographic angiogram performed in the primary evaluation of acute stroke without increasing the contrast dose. Twenty consecutive patients with acute ischemic stroke within the 4.5 hours of symptom onset were prospectively recruited. We increased the longitudinal coverage to the domes of the diaphragm to include the heart. Contrast administration (Omnipaque 350) remained unchanged (injected at 3-4 mL/s; total 60-80 mL, triggered by bolus tracking). Images of the heart and aorta, reconstructed at 5 mm slice thickness in 3 orthogonal planes, were read by a radiologist and cardiologist, findings conveyed to the treating neurologist, and correlated with the transthoracic or transesophageal echocardiogram performed within the next 24 hours. Of 20 patients studied, 3 (15%) had abnormal findings: a left ventricular thrombus, a Stanford type A aortic dissection, and a thrombus of the left atrial appendage. Both thrombi were confirmed by transesophageal echocardiography, and anticoagulation was started urgently the following day. None of the patients developed contrast-induced nephropathy on follow-up. The radiation dose was slightly increased from a mean of 4.26 mSV (range, 3.88-4.70 mSV) to 5.17 (range, 3.95 to 6.25 mSV). Including the heart and ascending aorta in a routine non-ECG-gated computed tomographic angiogram enhances an existing imaging modality, with no increased incidence of contrast-induced nephropathy and minimal increase in radiation dose. This may help in the detection of high-risk cardiac and aortic sources of embolism in acute stroke patients.


[Incidence of infected surgical wound and prophylaxis with cefotaxime in cesarean section].
Surgical wound infection after cesarean section varies from 2.5 to 16.1%, thus the utilization of antibiotic prophylaxis has increased routinely and irrationally. Despite this, we can still see cases of infections. To determine if the antibiotic prophylaxis with cefotaxime is associated with the decreased incidence of wound infection in patients submitted to cesarean section without risk factors. This study was carried out as a randomized clinical trial in patients submitted to cesarean section. Two groups were formed: in the first group we administered cefotaxime and the other one did not receive prophylaxis. The follow-up lasted 30 days to evaluate clinical data of infection. We performed 3,300 cesarean in the studied period; 1,000 patients had the inclusion criteria to participate in the study. A surgical wound infection was observed in 31 (0.96%) patients without risk factors. In 14 of these patients we administered cefotaxime, and in 17 patients we did not use prophylaxis. The highest frequency of infection was observed in the group of 24 to 30 years old, with 16 patients (51.6%). The clinical data of infection were: dehiscence in 29 patients (93.5%), pus secretion in 23 (74.2%), and fever in only 3 (9.7%) of them. The hospital stay after the infection was of five days in 75% of the cases. When the use of cefotaxime as prophylaxis was analyzed in both groups we had an odds ratio of 0.82, which was not significant. The use of cefotaxime in patients operated of cesarean does not have great transcendence since it does not reduce the infection incidence. The use of antibiotics in an irrational way implies a high cost, since the majority of the post-operation infections are not complicated, involving exclusively the skin and cellular subcutaneous tissue. Then, the cases with risk factors should be analyzed carefully for the cefotaxime administration.


Hepatoprotective immune response during Trichinella spiralis infection in mice.
Infections with gastrointestinal nematodes provoke immune and inflammatory responses mediated by cytokines released from T-helper type-2 (Th2) cells. Infections with Trichinella species have been reported to differ by the host species. Previously, in rats, we observed acute liver inflammation in response to infection with Trichinella spiralis, and the rat hosts showed a series of biochemical changes characterized by a decrease in serum paraoxonase (PON) 1 activity associated with the down-regulation of hepatic PON1 synthesis. In the present study, we investigated the effect(s) of species differences on the immune response against T. spiralis infection by analyzing serum PON1 activity and the associated inflammatory/anti-inflammatory mediators in mice. There were inconsistent changes in the serum PON1 activity of mice infected with T. spiralis, and these changes were associated with significant increases in the serum levels of interleukin (IL)-2, IL-4, IL-10, IL-12 (p70), granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor α during the enteric phase of the infection, while the levels of IL-5 and interferon γ were significantly increased throughout the entire experimental period. Moreover, T. spiralis infection in mice was associated with little inflammatory cell infiltration in hepatic tissues. Given the zoonotic prevalence of T. spiralis, further mechanistic research in this area is warranted.


SARS CoV-2 mRNA vaccination exposes latent HIV to Nef-specific CD8+ T-cells.
Efforts to cure HIV have focused on reactivating latent proviruses to enable elimination by CD8+ cytotoxic T-cells. Clinical studies of latency reversing agents (LRA) in antiretroviral therapy (ART)-treated individuals have shown increases in HIV transcription, but without reductions in virologic measures, or evidence that HIV-specific CD8+ T-cells were productively engaged. Here, we show that the SARS-CoV-2 mRNA vaccine BNT162b2 activates the RIG-I/TLR - TNF - NFκb axis, resulting in transcription of HIV proviruses with minimal perturbations of T-cell activation and host transcription. T-cells specific for the early gene-product HIV-Nef uniquely increased in frequency and acquired effector function (granzyme-B) in ART-treated individuals following SARS-CoV-2 mRNA vaccination. These parameters of CD8+ T-cell induction correlated with significant decreases in cell-associated HIV mRNA, suggesting killing or suppression of cells transcribing HIV. Thus, we report the observation of an intervention-induced reduction in a measure of HIV persistence, accompanied by precise immune correlates, in ART-suppressed individuals. However, we did not observe significant depletions of intact proviruses, underscoring challenges to achieving (or measuring) HIV reservoir reductions. Overall, our results support prioritizing the measurement of granzyme-B-producing Nef-specific responses in latency reversal studies and add impetus to developing HIV-targeted mRNA therapeutic vaccines that leverage built-in LRA activity.


[Protein and energy value of spiruline blue algae supplemented by amino acids: digestive and metabolic utilization by the growing rat].
Protein and energy value of 6 samples of "Spirulina" was studied on growing rats in 1972 and 1973. Sample RL 1(Spirulina platensis, originating from Tchad) was grown in artifical conditions in a laboratory. Others samples (Spirulina maxima) were grown in the solar evaporator near Mexico, washed and dried either on heated rollers (MR8, MR13) or by spraying (MA 7, MA10). Sample MA10 D corresponds to sample MA10, bleached by ethanol plus acetone (Baron, 1975). Each Spirulina sample was the only protein source of balanced, starch diets. The diets were supplemented in essential amino acids (E.A.A.) according to the requirements of growing rats (table 1). The ratios [(digestible nitrogen/metabolisable energy (EM] of the Spirulina diets were similar to that of the control diets containing herring meal. The diets were fed to groups of 15 to 17 growing rats. Energy and nitrogen balances were established by the comparative slaughter technique. Blood and muscle samples were taken at slaughter for the determination of free amino acids levels.


Detection of MDM2/CDK4 amplification in lipomatous soft tissue tumors from formalin-fixed, paraffin-embedded tissue: comparison of multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH).
In this study, the detection of MDM2 and CDK4 amplification was evaluated in lipomatous soft tissue tumors using multiplex ligation-dependent probe amplification (MLPA), a PCR-based technique, in comparison with fluorescence in situ hybridization (FISH). These 2 techniques were evaluated in a series of 77 formalin-fixed, paraffin-embedded lipomatous tumors (27 benign adipose tumors, 28 atypical lipomatous tumors/well-differentiated liposarcomas, 18 dedifferentiated liposarcomas, and 4 pleomorphic liposarcomas). Using MLPA, with a cut-off ratio of >2, 36/71 samples (22 atypical lipomatous tumors/well-differentiated liposarcomas, and 14 dedifferentiated liposarcomas) showed MDM2 and CDK4 amplification. Using FISH as gold standard, MLPA showed a sensitivity of 90% (36/40) and a specificity of 100% (31/31) in detecting amplification of MDM2 and CDK4 in lipomatous soft tissue tumors. In case of high-level amplification (MDM2-CDK4/CEP12 ratio >5), concordance was 100%. Four cases of atypical lipomatous tumor/well-differentiated liposarcoma (4/26, 15%) with a low MDM2 and CDK4 amplification level (MDM2-CDK4/CEP12 ratio ranging between 2 and 2.5) detected by FISH showed no amplification by MLPA, although gain of MDM2 and CDK4 (ratios ranging between 1.6 and 1.9) was seen with MLPA. No amplification was detected in benign lipomatous tumors and pleomorphic liposarcomas. Furthermore, there was a very high concordance between the ratios obtained by FISH and MLPA. In conclusion, MLPA proves to be an appropriate and straightforward technique for screening MDM2/CDK4 amplification in lipomatous tumors, especially when a correct cut-off value and reference samples are chosen, and could be considered a good alternative to FISH to determine MDM2 and CDK4 amplification in liposarcomas. Moreover, because MLPA, as a multiplex technique, allows simultaneous detection of multiple chromosomal changes of interest, it could be in the future a very reliable and fast molecular analysis on paraffin-embedded material to test for other diagnostically, prognostically, or therapeutically relevant genomic mutations in lipomatous tumors.


Bariatric surgery and gastroesophageal reflux disease.
With the rapidly increasing prevalence of obesity globally, the practice of bariatric surgery is being adopted routinely to prevent the development of chronic conditions as well as some forms of cancers associated with obesity. Gastroesophageal reflux disease (GERD) is one of those chronic conditions. Furthermore, there is accumulating data that obesity is associated with complications related to longstanding GERD such as erosive esophagitis (EE), Barrett's esophagus (BE), and esophageal adenocarcinoma (EAC). Central obesity, rather than body mass index (BMI), appears to be more closely associated with these complications. It should be expected, therefore, that weight loss procedures should result in improvement in GERD symptoms and its associated complications. However, in reality the different bariatric surgical procedures have unpredictable effects on an established GERD and may even produce GERD symptoms for the very first time (de novo). In this review, we explore the literature studying the effects of bariatric surgical operations on GERD. Roux-en-Y gastric bypass appears to have the most beneficial effect on GERD. On the other hand, laparoscopic sleeve gastrectomy and laparoscopic adjustable gastric banding (LAGB) are linked with long-term increased prevalence of GERD. We argue that GERD is an extremely important preoperative consideration for any patient undergoing bariatric surgery and therefore should be thoroughly investigated objectively (with 24-hour pH study and high-resolution manometry) to select the most suitable bariatric procedure for patients for their long-term success.


Components of human split ejaculates. II. Enzymes and proteinase inhibitors.
Lysozyme, alpha-amylase, neutral proteinase and plasminogen activator were most concentrated in the initial portion of the ejaculate that consists mostly of Cowper's gland and prostate gland fluids as well as spermatozoa. The concentration of the high molecular weight proteinase inhibitors, alpha1-antitrypsin and alpha1X-antichymotrypsin, was essentially unaltered throughout the ejaculate fractions, although their absolute amounts showed an increase towards the final fraction. By contrast, the total inhibitory activity towards pancreatic trypsin was highest both in concentration and amount in the last fraction, thus indicating that the seminal vesicles are its primary source. Plasminogen, prothrombin, Factor XIII, and the proteinase inhibitors antithrombin III, alpha2-macroglobulin, inter-alpha-trypsin inhibitor and C1S-inactivator could not be detected immunochemically in whole ejaculates, and indicates the dissimilarity between the coagulation/liquefaction processes of semen and blood.


BodiMojo: effective Internet-based promotion of positive body image in adolescent girls.
This study tested the efficacy of an Internet-based health promotion program, BodiMojo, designed to promote positive body image in adolescents. Participants were 178 students (mean age 15.2 years, 67.6% ethnic minority) in three public high schools. Intervention groups used BodiMojo for four weekly health class periods, while controls participated in their usual health curriculum. Body image measures were given at baseline, post-intervention, and 3 months. Girls reported decreased body dissatisfaction (p<.05), decreased physical appearance comparison (p<.05), and increased appearance satisfaction (p<.05), relative to controls. Effects were not maintained at 3-month follow-up. No significant differences were found between the intervention and control groups with boys. Moderation analyses suggested positive effects for diverse adolescents as well as those who were overweight or indicated baseline high body dissatisfaction. BodiMojo appears to be modestly effective in decreasing body image concerns among adolescent girls in the short term.


Conformational coupling between receptor and kinase binding sites through a conserved salt bridge in a signaling complex scaffold protein.
Bacterial chemotaxis is one of the best studied signal transduction pathways. CheW is a scaffold protein that mediates the association of the chemoreceptors and the CheA kinase in a ternary signaling complex. The effects of replacing conserved Arg62 of CheW with other residues suggested that the scaffold protein plays a more complex role than simply binding its partner proteins. Although R62A CheW had essentially the same affinity for chemoreceptors and CheA, cells expressing the mutant protein are impaired in chemotaxis. Using a combination of molecular dynamics simulations (MD), NMR spectroscopy, and circular dichroism (CD), we addressed the role of Arg62. Here we show that Arg62 forms a salt bridge with another highly conserved residue, Glu38. Although this interaction is unimportant for overall protein stability, it is essential to maintain the correct alignment of the chemoreceptor and kinase binding sites of CheW. Computational and experimental data suggest that the role of the salt bridge in maintaining the alignment of the two partner binding sites is fundamental to the function of the signaling complex but not to its assembly. We conclude that a key feature of CheW is to maintain the specific geometry between the two interaction sites required for its function as a scaffold.


Objective: Background regarding a recent debate between Cuijpers et al. (2019a, b) and the authors (Munder et al. 2019) about the efficacy of psychotherapy for depression is given. Method: A main reason for the discrepancy in Cuijpers et al.'s and our conclusions is discussed. Results: In our view the discrepancy is due, among other things, to a blurred distinction between questions of relative and absolute efficacy of psychotherapy. Although the efficacy of psychotherapy vis-à-vis alternative treatments may be ambiguous, there can be little doubt about the benefits of psychotherapy relative to no treatment. Conclusion: We do not think that raising fundamental concerns about the value of psychotherapy is a service to the field. We argue that moving the field forward requires a focus on how psychotherapy works and how the access to psychotherapy can be increased.


Neutrophil extracellular traps (NETs) and the role of platelets in infection.
In addition to playing a central role in normal haemostasis, platelets make important contributions to host inflammatory and immune responses to injury or infection. Under pathophysiological conditions where platelet function is not tightly controlled, platelets also play critical roles in pathogenic processes underlying cardiovascular disease, uncontrolled inflammation, coagulopathy and in tumour metastasis. Neutrophil extracellular traps (NETs) are webs of histone-modified nuclear material extruded from activated neutrophils during inflammatory responses and these degranulation events can be directly triggered by platelet/neutrophil engagement. Emerging research describes how NETs influence platelet function, particularly in the setting of infection and inflammation. Especially intriguing is the potential for platelet-driven coagulation to be modulated by NETs in plasma and interstitial spaces. These findings also reveal new perspectives related to improved therapy for venous thrombosis.


The Affordable Rental Housing Crisis and Population Health Equity: a Multidimensional and Multilevel Framework.
The US is facing a severe affordable rental housing crisis that contributes to multiple forms of housing insecurity including homelessness, crowded and poor quality housing conditions, unstable housing arrangements, and cost burdens. A considerable body of evidence finds that housing insecurity is an important determinant of health. However, the existing literature may fall short of conceptualizing and measuring the full impact of housing insecurity on population health and on racial health equity. In this paper, we seek to expand the conceptualization of housing as a determinant of population health equity by considering housing insecurity as a manifestation of structural racism that intersects with other manifestations and impacts of structural racism to affect, not only the health of housing insecure individuals, but also the health of the networks and communities in which these individuals live. First, we situate the current housing crisis within larger systems of structural racism. We extend prior work documenting the confluence of ways that racist policies and practices have created unequal burdens of housing insecurity to also discuss the ways that the meanings and impacts of housing insecurity may be shaped by racism. Next, we consider how the health impacts of this unequal burden of housing insecurity can extend beyond individual households to affect networks and communities. Ultimately, we provide a multilevel framework that can inform research, policy, and practice to address housing and health equity.


Capture and imaging of a prehairpin fusion intermediate of the paramyxovirus PIV5.
During cell entry, enveloped viruses fuse their viral membrane with a cellular membrane in a process driven by energetically favorable, large-scale conformational rearrangements of their fusion proteins. Structures of the pre- and postfusion states of the fusion proteins including paramyxovirus PIV5 F and influenza virus hemagglutinin suggest that this occurs via two intermediates. Following formation of an initial complex, the proteins structurally elongate, driving a hydrophobic N-terminal "fusion peptide" away from the protein surface into the target membrane. Paradoxically, this first conformation change moves the viral and cellular bilayers further apart. Next, the fusion proteins form a hairpin that drives the two membranes into close opposition. While the pre- and postfusion hairpin forms have been characterized crystallographically, the transiently extended prehairpin intermediate has not been visualized. To provide evidence for this extended intermediate we measured the interbilayer spacing of a paramyxovirus trapped in the process of fusing with solid-supported bilayers. A gold-labeled peptide that binds the prehairpin intermediate was used to stabilize and specifically image F-proteins in the prehairpin intermediate. The interbilayer spacing is precisely that predicted from a computational model of the prehairpin, providing strong evidence for its structure and functional role. Moreover, the F-proteins in the prehairpin conformation preferentially localize to a patch between the target and viral membranes, consistent with the fact that the formation of the prehairpin is triggered by local contacts between F- and neighboring viral receptor-binding proteins (HN) only when HN binds lipids in its target membrane.


The synthesis and evaluation of sesamol and benzodioxane derivatives as inhibitors of monoamine oxidase.
In the present study, series of eight sesamol (1,3-benzodioxol-5-ol) and eight benzodioxane (2,3-dihydro-1,4-benzodioxine) derivatives were synthesised and evaluated as inhibitors of recombinant human monoamine oxidase (MAO) A and B. The sesamol and benzodioxane derivatives are structurally related to series of phthalide derivatives, which have previously been found to act as potent reversible MAO inhibitors. The results document that the benzodioxane derivatives, in particular, are potent MAO-B inhibitors with IC50 values ranging from 0.045 to 0.947 μM. IC50 values for the inhibition of MAO-B by the homologous series of sesamol derivatives ranged from 0.164 to 7.29 μM. All compounds evaluated are selective for the MAO-B isoform, with IC50 values for the inhibition of MAO-A ranging from 13.2 to >100 μM. It is further shown that for the most potent MAO-B inhibitor, 6-[(3-bromophenyl)methoxy]-2,3-dihydro-1,4-benzodioxine, inhibition is almost completely reversed by dialysis of enzyme-inhibitor mixtures. It may be concluded that benzodioxane derivatives are promising leads for the design of selective MAO-B inhibitors for the treatment of Parkinson's disease.


[Characteristic of one-paired pea virus].
The new virus isolated from Vicia unijuga A.Br. with filament particles with size 1000-1200 x 10-12 nm is revealed. A thermal inactivation point is 55 degrees C; dilution end point - 10(-5)-10(-6) longevity in vitro in broad bean sap--less than one day. It is transferred by aphids and by pea, bean and broad bean seeds. The plants of Fabaceae, Solanaceae and Chenopodiaceae fam. were affected by this virus isolate. The virus yield was 40-50 mg per 100 g of leaves. The ratio of absorption E260/E280 corresponded to 1.4-1.5. The molecular mass of a core protein of the virus was 34 kD. The virus has a high immunogenic properties--titer is 1:256000 (indirect method of ELISA). It is presumably identified as a member of Closteroviridae.


Aerobic Exercise Recommendations to Optimize Best Practices in Care After Stroke: AEROBICS 2019 Update.
Most stroke survivors have very low levels of cardiovascular fitness, which limits mobility and leads to further physical deconditioning, increased sedentary behavior, and heightened risk of recurrent stroke. Although clinical guidelines recommend that aerobic exercise be a part of routine stroke rehabilitation, clinical uptake has been suboptimal. In 2013, an international group of stroke rehabilitation experts developed a user-friendly set of recommendations to guide screening and prescription-the Aerobic Exercise Recommendations to Optimize Best Practices in Care after Stroke (AEROBICS 2013). The objective of this project was to update AEROBICS 2013 using the highest quality of evidence currently available. The first step was to conduct a comprehensive review of literature from 2012 to 2018 related to aerobic exercise poststroke. A working group of the original consensus panel members drafted revisions based on synthesis. An iterative process was used to achieve agreement among all panel members. Final revisions included: (1) addition of 115 new references to replace or augment those in the original AEROBICS document, (2) rewording of the original recommendations and supporting material, and (3) addition of 2 new recommendations regarding prescription. The quality of evidence from which these recommendations were derived ranged from low to high. The AEROBICS 2019 Update should make it easier for clinicians to screen for, and prescribe, aerobic exercise in stroke rehabilitation. Clinical implementation will not only help to narrow the gap between evidence and practice but also reduce current variability and uncertainty regarding the role of aerobic exercise in recovery after stroke.


Validating a Minimally Invasive Tissue Sampling (MITS) Method in Determining Cause of Death in Stillbirths and Neonates.
Complete diagnostic autopsy (CDA) remains the gold standard and a valuable technique for determining cause of death. It is a source of health statistics that can be used to measure health care services' quality, unraveling important information on disease processes, particularly in emerging and unknown diseases. It can also be a vital tool for medical education and biomedical research. However, autopsy rates have been declining globally. There is an urgent need to develop and validate alternative methods in different settings to provide reliable information on cause of death. In this study, we aimed to determine cause of death (KazCoDe) in neonates and infants using minimally invasive tissue sampling (MITS), and to compare these results with those of CDA. We conducted MITS and CDA sequentially on 24 deceased children at the Pathological Bureau of the Akimat of the city of Nur-Sultan. Clinical data of the study subjects were extracted from their clinical records. During both procedures, brain, liver and lung tissues were collected for pathological diagnosis. Fifteen (62.5%) and nine (37.5%) were stillbirths and neonates, respectively. Eight (33.3%) were females and 16 (66.7%) were males. MITS diagnosis of cause of death was concordant with CDA diagnosis in 83.3% out of the 24 cases when considering the immediate and underlying causes of death and reviewing all the clinical and laboratory test results as part of the diagnostic evaluation to arrive at a cause of death (ICD-PM). We concluded that MITS is a valuable and reliable method for cause of death diagnosis in stillbirths and neonates, which can contribute vital mortality statistics in children in the absence of CDA.


Robust and efficient feature tracking for indoor navigation.
Robust feature tracking is a requirement for many computer vision tasks such as indoor robot navigation. However, indoor scenes are characterized by poorly localizable features. As a result, indoor feature tracking without artificial markers is challenging and remains an attractive problem. We propose to solve this problem by constraining the locations of a large number of nondistinctive features by several planar homographies which are strategically computed using distinctive features. We experimentally show the need for multiple homographies and propose an illumination-invariant local-optimization scheme for motion refinement. The use of a large number of nondistinctive features within the constraints imposed by planar homographies allows us to gain robustness. Also, the lesser computation cost in estimating these nondistinctive features helps to maintain the efficiency of the proposed method. Our local-optimization scheme produces subpixel accurate feature motion. As a result, we are able to achieve robust and accurate feature tracking.


Regulation of nitrogenase biosynthesis in Klebsiella pneumoniae: effect of nitrate.
The rate of biosynthesis of nitrogenase polypeptides in Klebsiella pneumoniae was determined in a medium containing NaNO3 or NaNO2. Nitrogenase biosynthesis was completely repressed by NO3- in a mutant strain, strain SK-25, that is derepressed for nitrogenase biosynthesis in the presence of NH4+. Chlorate-resistant mutants, derived from strain SK-25, that are defective in NO3- respiration produced nitrogenase in the presence of NO3-. Strain SK-56), a chlorate-resistant derivative capable of NO3- respiration, produced no nitrogenase in the presence of NO3- or NO2-. Klebsiella pneumoniae respired under anaerobic conditions utilizing either NO3- or NO2- as terminal electron acceptor. A mechanism for the control of nitrogenase biosynthesis is discussed involving the redox control of anaerobic enzyme systems.


The aberrant coexpression of several receptor tyrosine kinases is largely restricted to EBV-negative cases of classical Hodgkin's lymphoma.
The Hodgkin-Reed/Sternberg (HRS) cells of classical Hodgkin's lymphoma (HL) aberrantly express up to 7 different receptor tyrosine kinases (RTK) with extensive heterogeneity regarding the number and combinations of expressed RTKs in individual cases and a more prominent coexpression in nodular-sclerosis (ns) than mixed-cellularity (mc) HL. To investigate whether RTK expression patterns are related to other pathogenetic mechanisms and clinical behaviour, we analysed a large collection of EBV(+) and EBV(-) cases of ns and mc subtype and cases with relapses for expression of the 7 RTKs. No specific relation of any RTK to a specific group of cases was observed. The analysis of average numbers of expressed RTKs per case as a measure for strength of overall RTK signalling revealed a relation with the histological subtype and the EBV-status. RTK coexpression was significantly higher in EBV(-) nsHL cases compared to both EBV(-) and EBV(+) mcHL cases. Among mcHL cases RTK coexpression was significantly higher in EBV(-) compared to EBV(+) cases. Coexpression of 3 and more RTKs was largely restricted to EBV(-) cases. The inverse correlation between strong RTK signalling and presence of EBV may indicate that RTK signalling can at least partially replace the role of EBV in HRS cell pathogenesis. For cases with aberrant coexpression of several RTKs inclusion of RTK inhibitors in therapy regimens may be a novel option.


The relative effect of ascorbic acid on iron absorption from soy-based and milk-based infant formulas.
The effect of varying concentrations of ascorbic acid on the absorption of iron from a soy-based infant milk formula containing 6 mg iron/100 g was examined in 64 adult Indian females using the extrinsic radioactive tag method. The corrected geometric mean absorption from the basic soy formula was only 1.8%. Addition of ascorbic acid in a concentration of 40 mg/100 g, did not significantly increase absorption (3.3%; t = 1.8, p greater than 0.07) but raising the concentration to 80 mg/100 g did so (6.9%; t = 2.4, p less than 0.02). No further significant increase was noted when the concentration of ascorbic acid was increased to 160 mg/100 g (7.7%; t = 0.4, p greater than 0.7). The inhibitory effect of soy on iron absorption was further demonstrated by a direct comparison between the soy-based formula and a similar product based on cows' milk. The comparison was made at two concentrations of ascorbic acid. At 40 mg/100 g the geometric mean iron absorption from the soy formula was 2.4% compared with 5.3% from the milk formula (t = 2.8, p less than 0.02), while the corresponding values at 80 mg ascorbic acid/100 g were 7.2 and 19.5%, respectively (t = 3.4, p less than 0.02). The present results confirm the marked inhibitory effect of soy protein on iron absorption and calculations from the absorption figures suggest that such formulas should contain at least 12 mg/100 g iron together with ascorbic acid in a molar ratio of approximately 4:1 if they are to be adequate in terms of iron nutrition.


Tryparedoxin peroxidases and superoxide dismutases expression as well as ROS release are related to Trypanosoma cruzi epimastigotes growth phases.
Trypanosoma cruzi's antioxidant system is unique and relevant to the parasite. In this study, quantitative assays were performed to determine cytosolic and mitochondrial tryparedoxin peroxidases and superoxide dismutases expression (TcCPx, TcMPx, SODB and SODA) in correlation to H(2)O(2) release and O(2)(-) production. Differences were observed regarding H(2)O(2) release and O(2)(-) production between strains and along the growth curve. All of the enzymes studied exhibited varied expression as a function of time in culture. Although at lower levels, the Y strain exhibited the same pattern of Tulahuen 2 enzyme expression for all of the proteins studied, except SODA. In the stationary phase, the degree of expression of all of the enzymes in the Y strain returned to similar levels as those detected in the log phase with the exception of TcCPx and SODA. In Tulahuen 2, a higher expression of TcMPx, SODA and SODB was detected in the early stationary phase, and a slight decrease was observed in the late stationary phase for each enzyme, excluding TcMPx, which exhibited a marked decrease, and TcCPx, which increased its level. Because of the significance of ROS in redox signaling, these differences in enzyme expression underscore the importance of these parameters for epimastigote proliferation.


Food system dynamics structuring nutrition equity in racialized urban neighborhoods.
The food system is a social determinant of health and a leverage point for reducing diet-related racial inequities. Yet, food system interventions have not resulted in sustained improvement in dietary outcomes for underrepresented minorities living in neighborhoods with a history of disinvestment. Research is needed to illuminate the dynamics structuring food systems in racialized neighborhoods to inform intervention development. To conduct participatory research examining the complexity and inequity of food systems in historically redlined neighborhoods to identify feedback mechanisms to leverage in efforts to transform system outcomes for racial equity. We conducted a mixed-methods study in Cleveland, Ohio, USA from 2018 to 2021 using participatory system dynamic modeling with 30 academic and community partners, in-depth qualitative interviews with 22 key stakeholders, and public convenings with 250 local food policy council affiliates. Data were synthesized into causal loop diagrams depicting feedback mechanisms reinforcing or balancing neighborhood-level food system dynamics. We identified 10 feedback mechanisms structuring nutrition equity, which was identified as a meta-goal for food systems in racialized neighborhoods. Feedback mechanisms were organized in 3 domains: 1) meeting basic food needs with dignity (i.e., side hustle, government benefits, emergency food assistance, stigma, and stereotypes); 2) local food supply and demand dynamics (i.e., healthy food retail, job security, food culture, and norms); and 3) community empowerment and food sovereignty (i.e., community power, urban agriculture, risk of gentrification). Five exogenous factors moderate feedback dynamics: neighborhood crisis, neighborhood investments, household costs, government benefit funding, and voter participation. We identified nutrition equity as an overarching goal for local food systems, which reflects a state of having freedom, agency, and dignity in food traditions resulting in people and communities healthy in body, mind, and spirit. It is a transformative goal designed to spur system-level interventions that further racial equity through improved local food system dynamics.


Follow-up after intensive care: a single center study.
To study health problems, quality of life, functional status, and memory after intensive care. Adult patients ( n=346) discharged from a university hospital ICU. Prospective cohort study. Follow-up patients were found using the ICU database and the Peoples Registry. Quality of life (QOL) was measured with the Short Form 36 (SF-36) 6 months after ICU discharge. Semi-structured interviews, questionnaires, Glasgow Outcome Score (recovery), and Karnofsky Index (functional status) were used at consultations 7-8 months after ICU discharge. The SF-36 response rate was 64.5%, with scores significantly lower than population scores. Consultation patients ( n=136) did not differ from the rest ( n=210) regarding age, SAPS II scores, length of stay (LOS), and reasons for ICU admission. At follow-up 67.6% of consultation patients continued most activities, 75% looked after themselves, and 64.7% were non-workers, compared to 40.4% before the ICU admission. During and after the ICU stay, 40% lost more than 10 kg body weight. Fifty-eight (43%) could not remember anything from their ICU stay. At follow-up only 22 (16%) could remember having received information during their ICU stay. Three patients needed referral to other specialties. We should focus more on optimizing symptom management and giving repeated information after ICU discharge. Nutritional status and weight loss is another area of concern. More research is needed to find out how the broad range of psychosocial and physical problems following an ICU stay relates to the stay.


Changes in spores of Clostridium bifermentans caused by treatment with hydrogen peroxide and cations.
Spores of Clostridium bifermentans were treated with hydrogen peroxide until their peripheries had lost refractility. The centres of such spores only retained refractility at acid pH. Adding monovalent cations or increasing the pH caused the treated spores to lose their remaining refractility and decreased the turbidity of spore suspensions. Divalent cations prevented or reversed this loss of central refractility and decreased the fall in turbidity. Calcium ions also prevented but did not reverse the loss of central refractility which occurred on drying or applying pressure. Electron micrographs of spores treated with hydrogen peroxide showed that the cortex was depleted or absent and that the loss of central refractility was accompanied by protoplast swelling. It is suggested that divalent cations make spores resistant to drying and pressure by cross-linking negatively charged groups within the protoplast, and that together with hydrogen ions they neutralize the negatively charged groups, thus preventing the swelling of the protoplast, loss of refractility and fall in extinction which occur when divalent cations are replaced by monovalent cations.


Cytokines and oxygen radicals after hyperoxia in preterm and term alveolar macrophages.
To determine if the alveolar macrophage inflammatory cytokine response to oxygen differs in premature cells, macrophages were obtained from litters of premature (27 days) and term (31 days) rabbits. The majority of these cells were nonspecific esterase positive and actively phagocytosed latex particles. The cells that expressed cytokines also reacted with a monoclonal antibody against rabbit macrophages. After incubation overnight in 5 or 95% oxygen, the amount of interleukin (IL)-1beta and IL-8 mRNA was assessed by RT-PCR and the amount of cytokine protein by quantitative immunofluorescence microscopy. The preterm macrophage showed a significant increase in cytokine mRNA and protein after overnight incubation in 95% oxygen. This response was not seen in the term cells. Only premature macrophages had a significant increase in intracellular oxygen radical content, measured by 2',7'-dichlorofluorescin analysis, after incubation in 95% oxygen. This enhanced inflammatory cytokine response to oxygen may be one mechanism involved in the early development of chronic lung disease in premature infants.


A Controlled, Randomized, Double-blind Trial to Evaluate the Effect of Vegetables and Whole Grain Powder That Is Rich in Dietary Fibers on Bowel Functions and Defecation in Constipated Young Adults.
This study evaluated the effect of vege-powder (VP), mainly consisted of chicory, broccoli, and whole grains, on bowel habit improvement and constipation alleviation. Using the Roman standard II, 96 male and female subjects in their twenties with constipation symptoms were divided into a control group or VP group. Subjects in a control group were supplied with rice flakes-powder (RFP) and subjects in the VP group were provided with 30 g of VP twice daily for 4 weeks. Constipation relief effectiveness was surveyed on 5-point Likert scales depending on stool hardness, amount of stool, sensation of incomplete evacuation, and straining to defecate at day 0, 14, and 28 of RFP or VP intake. Repeated measures analysis of variance analysis revealed that VP intake caused significant temporal changes in stool hardness, amount, sensation of incomplete evacuation, and straining to defecate. In addition, significant differences between control and VP groups were found in stool hardness, amount, sensation of incomplete evacuation, and straining to defecate at day 14 and 28 of experimental diet consumption. VP supplement for 2 weeks significantly increased the evacuation frequency (1.04 ± 0.71), compared to control group (0.41 ± 0.64) and this increase was maintained at 4 week of diet supplements. This result showed that constipated subjects who consumed VP, mainly consisting of chicory, broccoli, and whole grains, improved constipation symptoms at 2 and 4 weeks of consumption compared to those of control group who were provided with RFP.


Preparation of glucoamylase microcapsule beads and application in solid-state fermentation.
Baijiu brewing adopts the solid-state fermentation method, using starchy raw materials, Jiuqu as saccharifying fermenting agent, and distilled spirits made by digestion, saccharification, fermentation and distillation. In the late stages of solid-state fermentation of Baijiu, the reduced activity of glucoamylase leads to higher residual starch content in the Jiupei, which affects the liquor yield. The direct addition of exogenous glucoamylase leads to problems such as the temperature of the fermentation environment rising too quickly, seriously affecting the growth of microorganisms. To solve the problem of reduced activity of glucoamylase in the late stage of solid-state fermentation of Baijiu, microcapsule beads (M-B) based on microcapsule emulsion were prepared and the effect of M-B on solid-state fermentation of Baijiu was investigated. The results showed that the release of M-B before and after drying was 53.27% and 25.77% in the liquid state (120 h) and 29.84% and 22.62% in the solid state (15 days), respectively. Adding M-B improved the alcohol by 0.33 %vol and reducing sugar content by 0.51%, reduced the residual starch content by 1.21% of the Jiupei, and had an insignificant effect on the moisture and acidity of the Jiupei. M-B have excellent sustained-release properties. The addition of M-B in solid-state fermentation significantly increased the alcohol content, reduced the residual starch content of Jiupei, ultimately improving the starch utilization rate and liquor yield of Baijiu brewing. The preparation of M-B provides methods and approaches for applying other active substances and microorganisms in the brewing of Baijiu. © 2023 Society of Chemical Industry.


Plastins: versatile modulators of actin organization in (patho)physiological cellular processes.
Many actin-binding proteins are expressed in eukaryotic cells. These polypeptides assist in stabilizing and rearranging the organization of the actin cytoskeleton in response to external stimuli, or during cell migration and adhesion. Here we review a particular set of actin-binding proteins called plastins. Plastins (also called fimbrins) belong to a subclass of actin-binding proteins known as actin bundling proteins. Three isoforms have been characterized in mammals: T-plastin is expressed in cells from solid tissue, whereas L-plastin occurs predominantly in hematopoietic cells. The third isoform, I-plastin, is specifically expressed in the small intestine, colon and kidney. These proteins share the unique property of cross-linking actin filaments into tight bundles. Although plastins are primarily involved in regulation of the actin cytoskeleton, they possess some unique features. For instance, they are implicated in invasion by pathogenic bacteria such as Shigella flexneri and Salmonella typhimurium. Also, L-plastin plays an important role in leukocyte function. T-plastin, on the other hand, is possibly involved in DNA repair. Finally, both T- and L-plastin are implicated in several diseases, and L-plastin is considered to be a valuable marker for cancer.


Safety of silastic sheet for orbital wall reconstruction.
Many implants are being used for the reconstruction of orbital wall fractures. The effect of the choice of implant for the reconstruction of an orbital wall fracture on the surgical outcome is under debate. The purpose of this article is to compare the outcomes of orbital wall reconstruction of small orbital wall fractures on the basis of the implants used. The authors conducted a retrospective study using electronic databases. Between March 2001 and December 2012, 461 patients with orbital wall fractures were included in this study. Among them, 431 patients in whom the fracture size was less than 300 mm(2) were analyzed. The fracture size was calculated using computed tomography scans of the orbit in the sagittal and coronal images. Cases in which the fracture size was less than 300 mm(2) were included in this study. One hundred and twenty-nine patients were treated with silastic sheets; 238 patients were treated with titanium meshes; and absorbable meshes were used in the case of 64 patients. Overall, 13 patients required revision, and the revision rate was 3.0%. The revision rate of the silastic sheet group was 5.4%. In the multivariable analysis, the revision rate of the group reconstructed with silastic sheets was highly statistically significant (P=0.043, odds ratio=3.65). However, other factors such as age, sex, fracture type, and fracture size were not significant. Reconstruction of orbital wall fractures with silastic sheets may cause more complications than that with other materials such as titanium meshes and absorbable meshes.


Long-term clinical performance of short 6-mm implants supporting single crowns in the posterior region: A 10-year cohort study.
Long-term clinical success of non-splinted, posterior, and short implants still is unclear. This prospective cohort study reports the 10-year follow-up of 6-mm implants supporting single crowns in the posterior region, and patient-reported outcomes. Baseline sample comprised 20 patients treated with 46 screw-retained crowns supported by 6-mm implants with moderately rough implant surface. Participants were recalled for a 10-year clinical follow-up to assess survival rates, biologic and mechanic conditions, quality of life (OHIP-14), and treatment satisfaction. Data were collected with clinical-radiographic exams and analyzed using descriptive and inferential statistics. Fourteen patients with 35 implant-crown units were examined after 127.6 ± 11.8 months. For the entire cohort period, 7/46 implants were lost (survival estimate: 77.7% at 133 months), and mechanic complications occurred in 14/46 units (survival estimate: 66.4% at 116 months). In the Cox models, "maximum occlusal force" had a significant effect for implant loss (p = 0.038) and for prosthetic screw loosening (p = 0.038); "arch" and "bruxism" were not significant. Peri-implant bone loss was 0.4 ± 0.6 mm at 10 years. For peri-implant bone level, "crown-to-implant ratio" (p < 0.001) and "time" (p = 0.001) were significant. Bone levels differed from baseline to 12, 48, and 120 months. Satisfaction VAS was 94.0 ± 7.9 mm and OHIP-14 was 2.3 ± 2.2. Single screw-retained crowns supported by 6-mm implants have an acceptable long-term clinical performance, with stable peri-implant bone levels after 10 years of function.


Task-free spectral EEG dynamics track and predict patient recovery from severe acquired brain injury.
For some patients, coma is followed by a state of unresponsiveness, while other patients develop signs of awareness. In practice, detecting signs of awareness may be hindered by possible impairments in the patient's motoric, sensory, or cognitive abilities, resulting in a substantial proportion of misdiagnosed disorders of consciousness. Task-free paradigms that are independent of the patient's sensorimotor and neurocognitive abilities may offer a solution to this challenge. A limitation of previous research is that the large majority of studies on the pathophysiological processes underlying disorders of consciousness have been conducted using cross-sectional designs. Here, we present a study in which we acquired a total of 74 longitudinal task-free EEG measurements from 16 patients (aged 6-22 years, 12 male) suffering from severe acquired brain injury, and an additional 16 age- and education-matched control participants. We examined changes in amplitude and connectivity metrics of oscillatory brain activity within patients across their recovery. Moreover, we applied multi-class linear discriminant analysis to assess the potential diagnostic and prognostic utility of amplitude and connectivity metrics at the individual-patient level. We found that over the course of their recovery, patients exhibited nonlinear frequency band-specific changes in spectral amplitude and connectivity metrics, changes that aligned well with the metrics' frequency band-specific diagnostic value. Strikingly, connectivity during a single task-free EEG measurement predicted the level of patient recovery approximately 3 months later with 75% accuracy. Our findings show that spectral amplitude and connectivity track patient recovery in a longitudinal fashion, and these metrics are robust pathophysiological markers that can be used for the automated diagnosis and prognosis of disorders of consciousness. These metrics can be acquired inexpensively at bedside, and are fully independent of the patient's neurocognitive abilities. Lastly, our findings tentatively suggest that the relative preservation of thalamo-cortico-thalamic interactions may predict the later reemergence of awareness, and could thus shed new light on the pathophysiological processes that underlie disorders of consciousness.


Association Between Individual and Intimate Partner Factors and Cervical Cancer Screening in Kenya.
Cervical cancer is the most prevalent cancer among women in Kenya. Although cervical cancer screening could reduce illness and death, screening rates remain low. Kenyan women's individual characteristics and intimate partner factors may be associated with cervical cancer screening; however, a lack of nationally representative data has precluded study until recently. The objective of our study was to examine individual and intimate partner factors associated with cervical cancer screening in Kenya. We conducted secondary data analysis of responses by women who completed the cervical cancer screening and domestic violence questions in the Kenya Demographic and Health Survey, 2014 (N = 3,222). By using multivariable regression analyses, we calculated the association of cervical cancer screening with age, religion, education, wealth, recent exposure to family planning on television, head of household's sex, and experience of intimate partner violence. Rates of cervical cancer screening among women in Kenya increased with age. The wealthiest women and women with post-secondary education had greater odds of reporting being screened for cervical cancer than the poorest women and uneducated women. Christians and women exposed to prevention messaging on television had higher odds of screening than Muslims and women with no exposure. Victims of intimate partner violence had lower odds of being screened than women who had not experienced intimate partner violence. Identified barriers to screening in this sample mirror previous findings, though with additional nuances. Model fit data and theoretical review suggest that additional, unmeasured variables may contribute to variability in cervical cancer screening rates. Inclusion of additional variables specific to cervical cancer in future national surveys could strengthen the ability to identify factors associated with screening.


Unexpected Discovery of Dichloroacetate Derived Adenosine Triphosphate Competitors Targeting Pyruvate Dehydrogenase Kinase To Inhibit Cancer Proliferation.
Pyruvate dehydrogenase kinases (PDKs) have recently emerged as an attractive target for cancer therapy. Herein, we prepared a series of compounds derived from dichloroacetate (DCA) which inhibited cancer cells proliferation. For the first time, we have successfully developed DCA derived inhibitors that preferentially bind to the adenosine triphosphate (ATP) pocket of PDK isoform 1 (PDK1).


Transactional eHealth Literacy: Developing and Testing a Multi-Dimensional Instrument.
Theoretically informed measures of eHealth literacy that consider the social affordances of eHealth are limited. This study describes the psychometric testing of a multi-dimensional instrument to measure functional, communicative, critical, and translational eHealth literacies, as informed by the Transactional Model of eHealth Literacy (TMeHL). A 3-phase rating scale construction process was conducted to engage eHealth experts and end-users. In Phase 1, Experts (N = 5) and end-users (N = 25) identified operational behaviors to measure each eHealth literacy dimension. End-users (N = 10) participated in think-aloud interviews to provide feedback on items reviewed and approved by experts. A field test was conducted with a random sample of patients recruited from a university-based research registry (N = 283). Factor analyses and Rasch procedures examined the internal structure of the scores produced by each scale. Pearson's r correlations provided evidence for external validity of scores. The instrument measures four reliable (ω = .92-.96) and correlated (r= .44-.64) factors: functional (4 items), communicative (5 items), critical (5 items), and translational (4 items). Researchers and providers can use this new instrument as a theory-driven instrument to measure four eHealth literacies that are fundamental to the social affordances of the eHealth experience.


Cell-cycle-dependent regulation of androgen receptor function.
The androgen receptor (AR) is a critical oncogene in prostate cancer (PCa) development and progression. In this study, we demonstrate cell-cycle-dependent regulation of AR activity, localization, and phosphorylation. We show that for three AR-target genes, androgen-stimulated AR transactivation is highest during the G1 phase, decreased during S-phase, and abrogated during G2/M. This change in AR transactivation parallels changes in AR localization and phosphorylation. A combination of imaging techniques and quantitative analysis reveals nuclear AR localization during interphase and the exclusion of the majority, but not all, AR from chromatin during mitosis. Flow cytometry analyses using a phospho-S308 AR-specific antibody in asynchronous and chemically enriched G2/M PCa cells revealed ligand-independent induction of S308 phosphorylation in mitosis when CDK1 is activated. Consistent with our flow cytometry data, IP-western blotting revealed an increase in S308 phosphorylation in G2/M, and the results of an in vitro kinase assay indicated that CDK1 was able to phosphorylate the AR on S308. Pharmacological inhibition of CDK1 activity resulted in decreased S308 phosphorylation in PCa cells. Importantly, using a combination of anti-total AR and phospho-S308-specific antibodies in immunofluorescence experiments, we showed that the AR is excluded from condensed chromatin in mitotic cells when it was phosphorylated on S308. In summary, we show that the phosphorylation of the AR on S308 by CDK1 during mitosis regulates AR localization and correlates with changes in AR transcriptional activity. These findings have important implications for understanding the function of AR as an oncogene.


Frequent epigenetic silencing of protocadherin 10 by methylation in multiple haematologic malignancies.
Epigenetic silencing of tumour suppressor genes (TSG) inactivates TSG functions. Previously, we identified PCDH10 as a methylated TSG in carcinomas. Here, we detected its frequent silencing and methylation in lymphoma cell lines including 100% Burkitt, 100% diffuse large B cell, 86% Hodgkin, 100% nasal natural killer/T-cell lymphoma and 1/3 of leukaemia cell lines, and in primary tumours but not in normal peripheral blood mononuclear cells or lymph nodes. PCDH10 silencing could be reversed by demethylation with 5-aza-2'-deoxycytidine. Methylation was further detected in 14% of Hodgkin lymphoma sera. Thus, PCDH10 methylation is frequently involved in lymphomagenesis and could serve as a tumour-specific biomarker.


The rostral migratory stream plays a key role in intranasal delivery of drugs into the CNS.
The blood brain barrier (BBB) is impermeable to most drugs, impeding the establishment of novel neuroprotective therapies and strategies for many neurological diseases. Intranasal administration offers an alternative path for efficient drug delivery into the CNS. So far, the anatomical structures discussed to be involved in the transport of intranasally administered drugs into the CNS include the trigeminal nerve, olfactory nerve and the rostral migratory stream (RMS), but the relative contributions are debated. In the present study we demonstrate that surgical transection, and the resulting structural disruption of the RMS, in mice effectively obstructs the uptake of intranasally administered radioligands into the CNS. Furthermore, using a fluorescent cell tracer, we demonstrate that intranasal administration in mice allows agents to be distributed throughout the entire brain, including olfactory bulb, hippocampus, cortex and cerebellum. This study provides evidence of the vital role the RMS has in the CNS delivery of intranasally administered agents. The identification of the RMS as the major access path for intranasally administered drugs into the CNS may contribute to the development of treatments that are tailored for efficient transport within this structure. Research into the RMS needs to continue to elucidate its limitations, capabilities, mechanisms of transport and potential hazards before we are able to advance this technique into human research.


A multinational study of colonization with extended spectrum β-lactamase-producing Enterobacteriaceae in healthcare personnel and family members of carrier patients hospitalized in rehabilitation centres.
The study aims were: (i) to define the prevalence of and risk factors for colonization by extended spectrum β-lactamase (ESBL) -producing Enterobacteriaceae (EPE) among healthcare workers (HCWs) and family members (FMs) of EPE-colonized patients in rehabilitation units and (ii) to compare EPE isolates from these three groups. The study included 286 FMs of 194 EPE-carrying patients identified in five rehabilitation units located in Israel, Italy, France and Spain. The EPE were detected in rectal swabs from 26 (9%) of 286 FMs screened. In multivariate analyses, older age of FM, greater mean number of hours spent with the patient, being a daughter or a female spouse of a patient, and chronic lung disease of the patient were significantly associated with carriage in the FM. Escherichia coli was the most common organism (76%), followed by Klebsiella pneumoniae (19%). Isolates were typed by pulsed field gel electrophoresis and multilocus sequence typing, and ESBLs were identified by PCR sequencing. A comparison of paired species isolates from FMs and their respective patient showed that 17 of 23 strains were indistinguishable. EPE were detected in 35 (3.5%, E. coli = 34) of the 1001 HCWs screened. Feeding patients was associated with EPE carriage by HCWs. Only 7 of 23 E. coli subclones cultured from HCWs were also represented among 376 patient-derived ESBL-producing E. coli isolates from the same rehabilitation units. In Spain, a higher proportion of HCWs and FMs were ESBL carriers than elsewhere (p <0.05). In conclusion, the molecular and epidemiological data suggest that FMs are at higher risk of EPE acquisition from their relative patients than HCWs.


Indolcarboxamide is a preclinical candidate for treating multidrug-resistant tuberculosis.
New chemotherapeutic compounds against multidrug-resistant Mycobacterium tuberculosis (Mtb) are urgently needed to combat drug resistance in tuberculosis (TB). We have identified and characterized the indolcarboxamides as a new class of antitubercular bactericidal agent. Genetic and lipid profiling studies identified the likely molecular target of indolcarboxamides as MmpL3, a transporter of trehalose monomycolate that is essential for mycobacterial cell wall biosynthesis. Two lead candidates, NITD-304 and NITD-349, showed potent activity against both drug-sensitive and multidrug-resistant clinical isolates of Mtb. Promising pharmacokinetic profiles of both compounds after oral dosing in several species enabled further evaluation for efficacy and safety. NITD-304 and NITD-349 were efficacious in treating both acute and chronic Mtb infections in mouse efficacy models. Furthermore, dosing of NITD-304 and NITD-349 for 2 weeks in exploratory rat toxicology studies revealed a promising safety margin. Finally, neither compound inhibited the activity of major cytochrome P-450 enzymes or the hERG (human ether-a-go-go related gene) channel. These results suggest that NITD-304 and NITD-349 should undergo further development as a potential treatment for multidrug-resistant TB.


Midlevel Providers Focusing on Geriatrics Improve Care and Outcomes of Fall-Related Injuries Among the Elderly.
A rural level 1 trauma center underwent a consolidation to level III status in a new trauma network system. A dedicated group of midlevel practitioners emphasizing early mobilization, a geriatric care model, and fall prevention replaced surgical residents in the level 3 center. We hypothesized that outcomes of elderly fall-related injuries may be enhanced with midlevel providers using a geriatric-focused care model. An IRB-approved trauma registry review of patients over 65 years of age with a fall-related injury admitted to a rural trauma center 1 year prior to and 1 year following a trauma center consolidation from level 1 to level III designation evaluated demographics, anticoagulant use, comorbidities, and clinical outcomes. Statistical analysis included t-test and regression analysis. 327 patients injured by falls were seen over a 2-year study period. The number of patients admitted with a fall-related injury and the injury severity were similar over the study period. Increasing age and anticoagulant use increased length of stay and mortality (both with P < .05). Mortality rates and patient level of independence on discharge were improved in the later period involving midlevel practitioners (both with P < .05). Trauma centers and trauma system networks face increasing challenges to provide resources and providers of care for patients injured by falls, especially for the growing elderly population. Midlevel providers focusing on geriatric clinical issues and goals may enhance care and outcomes of elderly fall-related injuries.


Human microbiome and HIV/AIDS.
Understanding of the human microbiome continues to grow rapidly; however, reports on changes in the microbiome after HIV infection are still limited. This review surveys the progress made in methodology associated with microbiome studies and highlights the remaining challenges to this field. Studies have shown that commensal oral, gut, vaginal, and penile bacteria are vital to the health of the human immune system. Our studies on crosstalk among oral and gastrointestinal soluble innate factors, HIV, and microbes indicated that the oral and gut microbiome was altered in the HIV-positive samples compared to the negative controls. The importance of understanding the bacterial component of HIV/AIDS, and likelihood of "crosstalk" between viral and bacterial pathogens, will help in understanding the role of the microbiome in HIV-infected individuals and facilitate identification of novel antiretroviral factors for use as novel diagnostics, microbicides, or therapeutics against HIV infection.