The Sauvé-Kapandji procedure for posttraumatic wrist disorders: further experience.
A prospective survey was conducted to evaluate the outcome of the Sauvé-Kapandji procedure for posttraumatic wrist disorders. Eighty four patients were treated, all with posttraumatic disorders of the distal radioulnar joint, 73 as an isolated procedure, 11 in combination with another wrist procedure. There was significant pain decrease and high patient satisfaction (74%). The range of motion increased in the flexion/extension arc from 109 degrees to 124 degrees (p = 0.006) and, in those with limited forearm rotation, from 71 degrees to 134 degrees (p = 0.006). According to the Mayo Clinic wrist score, we obtained 20 excellent, 34 good, 18 fair and 12 poor results. Complications were rare.


Multimodal imaging: an evaluation of univariate and multivariate methods for simultaneous EEG/fMRI.
The combination of electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) has been proposed as a tool to study brain dynamics with both high temporal and high spatial resolution. Multimodal imaging techniques rely on the assumption of a common neuronal source for the different recorded signals. In order to maximally exploit the combination of these techniques, one needs to understand the coupling (i.e., the relation) between electroencephalographic (EEG) and fMRI blood oxygen level-dependent (BOLD) signals. Recently, simultaneous EEG-fMRI measurements have been used to investigate the relation between the two signals. Previous attempts at the analysis of simultaneous EEG-fMRI data reported significant correlations between regional BOLD activations and modulation of both event-related potential (ERP) and oscillatory EEG power, mostly in the alpha but also in other frequency bands. Beyond the correlation of the two measured brain signals, the relevant issue we address here is the ability of predicting the signal in one modality using information from the other modality. Using multivariate machine learning-based regression, we show how it is possible to predict EEG power oscillations from simultaneously acquired fMRI data during an eyes-open/eyes-closed task using either the original channels or the underlying cortically distributed sources as the relevant EEG signal for the analysis of multimodal data.


Hierarchical amplitude modulation structures and rhythm patterns: Comparing Western musical genres, song, and nature sounds to Babytalk.
Statistical learning of physical stimulus characteristics is important for the development of cognitive systems like language and music. Rhythm patterns are a core component of both systems, and rhythm is key to language acquisition by infants. Accordingly, the physical stimulus characteristics that yield speech rhythm in "Babytalk" may also describe the hierarchical rhythmic relationships that characterize human music and song. Computational modelling of the amplitude envelope of "Babytalk" (infant-directed speech, IDS) using a demodulation approach (Spectral-Amplitude Modulation Phase Hierarchy model, S-AMPH) can describe these characteristics. S-AMPH modelling of Babytalk has shown previously that bands of amplitude modulations (AMs) at different temporal rates and their phase relations help to create its structured inherent rhythms. Additionally, S-AMPH modelling of children's nursery rhymes shows that different rhythm patterns (trochaic, iambic, dactylic) depend on the phase relations between AM bands centred on ~2 Hz and ~5 Hz. The importance of these AM phase relations was confirmed via a second demodulation approach (PAD, Probabilistic Amplitude Demodulation). Here we apply both S-AMPH and PAD to demodulate the amplitude envelopes of Western musical genres and songs. Quasi-rhythmic and non-human sounds found in nature (birdsong, rain, wind) were utilized for control analyses. We expected that the physical stimulus characteristics in human music and song from an AM perspective would match those of IDS. Given prior speech-based analyses, we also expected that AM cycles derived from the modelling may identify musical units like crotchets, quavers and demi-quavers. Both models revealed an hierarchically-nested AM modulation structure for music and song, but not nature sounds. This AM modulation structure for music and song matched IDS. Both models also generated systematic AM cycles yielding musical units like crotchets and quavers. Both music and language are created by humans and shaped by culture. Acoustic rhythm in IDS and music appears to depend on many of the same physical characteristics, facilitating learning.


The use of thermosensitive quartz sensor for thermal regulation at cryogenic temperatures: application to microwave sapphire resonator references.
We demonstrated the use of thermosensitive quartz resonator oscillator as a thermal sensor for temperature control at the liquid nitrogen temperature. The high sensitivity of the quartz enables an efficient thermal regulation at ambient temperature as well as liquid nitrogen temperature. LC-cut quartz oscillator phase noise measurements show that the temperature measurement resolution is not limited by the intrinsic noise of the sensor and that a resolution of 10 muK can be achieved. This thermal regulation is applied to control a microwave temperature-compensated sapphire resonator oscillator at a temperature above 77 K, enabling the achievement of a flicker floor of 9.10(-13 ) at 9 GHz.


Comparison of the performance of different discriminant algorithms in analyte discrimination tasks using an array of carbon black--polymer composite vapor detectors.
An array of 20 compositionally different carbon black--polymer composite chemiresistor vapor detectors was challenged under laboratory conditions to discriminate between a pair of extremely similar pure analytes (H2O and D2O), compositionally similar mixtures of pairs of compounds, and low concentrations of vapors of similar chemicals. Several discriminant algorithms were utilized, including k nearest neighbors (kNN, with k = 1), linear discriminant analysis (LDA, or Fisher's linear discriminant), quadratic discriminant analysis (QDA), regularized discriminant analysis (RDA, a hybrid of LDA and QDA), partial least squares, and soft independent modeling of class analogy (SIMCA). H2O and D2O were perfectly classified by most of the discriminants when a separate training and test set was used. As expected, discrimination performance decreased as the analyte concentration decreased, and performance decreased as the composition of the analyte mixtures became more similar. RDA was the overall best-performing discriminant, and LDA was the best-performing discriminant that did not require several cross-validations for optimization.


The Potential Impact of Social Genomics on Wound Healing.
Significance: Human skin wounds carry an immense epidemiologic and financial burden, and their impact will continue to grow with an aging population and rising incidence of comorbid conditions known to affect wound healing. To comprehensively address this growing clinical issue, physicians should also be aware of how conditions of the human social environment may affect wound healing. Here we provide a review of the emerging field of social genomics and its potential impact on the wound healing. Recent Advances: Multiple studies using human and animal models have correlated social influences and their contributing effects to acute and chronic stress with delays in wound healing. Furthermore, observations between nongenetic factors such as nutrition, socioeconomic, and educational status have also shown to have a direct or indirect impact on clinical outcomes of wound healing. Critical Issues: Nutrition, financial burden, socioeconomic and education status, and acute and chronic stress are variables that have either direct (epigenetic) or indirect impact on wound healing and patients' quality of life. Wound care is costly and remains a challenge placing economic burden on patients. Furthermore, poor clinical outcomes and complications including loss of mobility and disability may lead to job loss, further contributing to socioeconomic related stress. Thus, the economic burden and inadequate wound healing are intertwined, making each other worse. Future Directions: Although some evidence regarding the specific changes in genetic pathways imparted by conditions of the social environment exists, further studies are warranted to identify potential mechanisms, interventions, and prevention approaches.


Mice deficient in protein tyrosine phosphatase receptor type Z (PTPRZ) show reduced responsivity to methamphetamine despite an enhanced response to novelty.
Methamphetamine (METH), a commonly abused drug, elevates extracellular dopamine (DA) levels by inducing DA efflux through the DA transporter (DAT). Emerging evidence in rodent models suggests that locomotor responses to a novel inescapable open field may predict behavioral responses to abused drugs; METH produces more potent stimulant effects in high responders to novelty than in low responders. We herein found that mice deficient in protein tyrosine phosphatase receptor type Z (Ptprz-KO) exhibited an enhanced behavioral response to novelty; however, METH-induced hyperlocomotion was significantly lower in Ptprz-KO than in wild-type mice when METH was administered at a non-toxic dose of 1 mg per kg body weight (bdw). Single-cell RT-PCR revealed that the majority of midbrain DA neurons expressed PTPRZ. No histological alterations were observed in the mesolimbic or nigrostriatal dopaminergic pathways in Ptprz-KO brains; however, a significant decrease was noted in brain DA turnover, suggesting functional alterations. In vivo microdialysis experiments revealed that METH-evoked DA release in the nucleus accumbens was significantly lower in Ptprz-KO mice than in wild-type mice. Consistent with this result, Ptprz-KO mice showed significantly fewer cell surface DAT as well as weaker DA uptake activity in striatal synaptosomes prepared 1 hr after the administration of METH than wild-type mice, while no significant differences were observed in the two groups treated with saline. These results indicate that the high response phenotype of Ptprz-KO mice to novelty may not be simply attributed to hyper-dopaminergic activity, and that deficits in PTPRZ reduce the effects of METH by reducing DAT activity.


PredictONCO: a web tool supporting decision-making in precision oncology by extending the bioinformatics predictions with advanced computing and machine learning.
PredictONCO 1.0 is a unique web server that analyzes effects of mutations on proteins frequently altered in various cancer types. The server can assess the impact of mutations on the protein sequential and structural properties and apply a virtual screening to identify potential inhibitors that could be used as a highly individualized therapeutic approach, possibly based on the drug repurposing. PredictONCO integrates predictive algorithms and state-of-the-art computational tools combined with information from established databases. The user interface was carefully designed for the target specialists in precision oncology, molecular pathology, clinical genetics and clinical sciences. The tool summarizes the effect of the mutation on protein stability and function and currently covers 44 common oncological targets. The binding affinities of Food and Drug Administration/ European Medicines Agency -approved drugs with the wild-type and mutant proteins are calculated to facilitate treatment decisions. The reliability of predictions was confirmed against 108 clinically validated mutations. The server provides a fast and compact output, ideal for the often time-sensitive decision-making process in oncology. Three use cases of missense mutations, (i) K22A in cyclin-dependent kinase 4 identified in melanoma, (ii) E1197K mutation in anaplastic lymphoma kinase 4 identified in lung carcinoma and (iii) V765A mutation in epidermal growth factor receptor in a patient with congenital mismatch repair deficiency highlight how the tool can increase levels of confidence regarding the pathogenicity of the variants and identify the most effective inhibitors. The server is available at https://loschmidt.chemi.muni.cz/predictonco.


Mitral valve replacement in severely calcified mitral valve annulus: a 10-year experience.
Severe calcification in the mitral valve annulus is a challenging problem during mitral valve surgery. We describe our experience with mitral valve replacement in severely calcified mitral valve without decalcification of the annulus. Between April 2001 and July 2011, 61 patients underwent mitral valve replacement with severe mitral annulus calcification without decalcification of the annulus. This retrospective study was performed to assess the surgical and the long-term postoperative outcomes in this group. The mean age of the patients was 75.2 ± 9.2 years. Twenty-four patients (53%) were in New York Heart Association Class III/IV. Twenty-six patients (58%) had good left ventricular function. Mean logistic EuroSCORE was 8.75. Isolated mitral valve replacement was performed in 12 patients (27%). Coronary artery bypass grafting was done in 13 patients (29%). In-hospital mortality was 4.9% (3 patients). Postoperative morbidity included re-exploration for bleeding in 3 patients (7%) and transient renal impairment in 10 patients (22%). Three patients required intra-aortic balloon pump (7%) for low cardiac output syndrome. Seven patients (16%) required permanent pacemaker, and 1 patient (2%) had thromboembolic event. The 1-year survival was 93.3%, and the 5-year survival was 78.8%. The mean echocardiography follow-up was 40 months. There was no paravalvular leak detected in any patient in the long-term follow-up. None of the patients had valve-related reoperation. Mitral valve replacement without annular decalcification in severely calcified mitral valve annulus is a safe and an effective approach and has good long-term outcome.


Enterostatin suppresses food intake in rats after near-celiac and intracarotid arterial injection.
Enterostatin (Ent) selectively suppresses the intake of dietary fat after peripheral and central administration. To further investigate the site of action of Ent, we compared the feeding responses to Ent injected intra-arterially near the celiac artery, into the carotid artery, or intravenously in rats adapted to a high-fat diet. After near-celiac arterial injection there was an immediate dose-dependent (0.05-13.5 nmol) inhibition of food intake occurring within 5 min in overnight-fasted rats that lasted up to 20 min. Carotid arterial Ent had a similar, immediate dose-related response, and the inhibitory effect was long lasting. The response to intravenous Ent was only evident at the highest dose (13.5 nmol) and was delayed for at least 120 min. Pretreatment with capsaicin, which causes degeneration of vagal sensory neurons, abolished the inhibitory responses to near-celiac Ent but not to intravenous or intracarotid Ent. These results provide further evidence for both a gastrointestinal site of action for peripheral Ent and a central site of action for intracarotid Ent and suggest that the delayed response to intravenous Ent may reflect either binding or slow uptake of this peptide into the central nervous system.


Pathological response of cavernous malformations following radiosurgery.
Stereotactic radiosurgery (SRS) is a therapeutic option for repeatedly hemorrhagic cavernous malformations (CMs) located in areas deemed to be high risk for resection. During the latency period of 2 or more years after SRS, recurrent hemorrhage remains a persistent risk until the obliterative process has finished. The pathological response to SRS has been studied in relatively few patients. The authors of the present study aimed to gain insight into the effect of SRS on CM and to propose possible mechanisms leading to recurrent hemorrhages following SRS. During a 13-year interval between 2001 and 2013, bleeding recurred in 9 patients with CMs that had been treated using Gamma Knife surgery at the authors' institution. Microsurgical removal was subsequently performed in 5 of these patients, who had recurrent hemorrhages between 4 months and 7 years after SRS. Specimens from 4 patients were available for analysis and used for this report. Histopathological analysis demonstrated that vascular sclerosis develops as early as 4 months after SRS. In the samples from 2 to 7 years after SRS, sclerotic vessels were prominent, but there were also vessels with incomplete sclerosis as well as some foci of neovascularization. Recurrent bleeding after SRS for CM could be related to incomplete sclerosis of the vessels, but neovascularization may also play a role.


Effects of ingesting supplements designed to promote lean tissue accretion on body composition during resistance training.
This study examined the effects of ingesting nutritional supplements designed to promote lean tissue accretion on body composition alterations during resistance training. Twenty-eight resistance-trained males blindly supplemented their diets with maltodextrin (M), Gainers Fuel 1000 (GF), or Phosphagain (P). No significant differences were observed in absolute or relative total body water among groups. Energy intake and body weight significantly increased in all groups combined throughout the study with no group or interaction differences observed. Dual energy x-ray absorptiometry-determined body mass significantly increased in each group throughout the study with significantly greater gains observed in the GF and P groups. Lean tissue mass (excluding bone) gain was significantly greater in the P group, while fat mass and percent body fat were significantly increased in the GF group. Results indicate that total body weight significantly increased in each group and that P supplementation resulted in significantly greater gains in lean tissue mass during resistance training.


Ambulatory Intensive Care for Medically Complex Patients at a Health Care Clinic for Individuals Experiencing Homelessness: The SUMMIT Randomized Clinical Trial.
Intensive primary care interventions have been promoted to reduce hospitalization rates and improve health outcomes for medically complex patients, but evidence of their efficacy is limited. To assess the efficacy of a multidisciplinary ambulatory intensive care unit (A-ICU) intervention on health care utilization and patient-reported outcomes. The Streamlined Unified Meaningfully Managed Interdisciplinary Team (SUMMIT) randomized clinical trial used a wait-list control design and was conducted at a health care clinic for patients experiencing homelessness in Portland, Oregon. The first patient was enrolled in August 2016, and the last patient was enrolled in November 2019. Included patients had 1 or more hospitalizations in the prior 6 months and 2 or more chronic medical conditions, substance use disorder, or mental illness. Data analysis was performed between March and May 2021. The A-ICU included a team manager, a pharmacist, a nurse, care coordinators, social workers, and physicians. Activities included comprehensive 90-minute intake, transitional care coordination, and flexible appointments, with reduced panel size. Enhanced usual care (EUC), consisting of team-based primary care with access to community health workers and mental health, addiction treatment, and pharmacy services, served as the comparator. Participants who received EUC joined the A-ICU intervention after 6 months. The main outcome was the difference in rates of hospitalization (primary outcome), emergency department (ED) visits, and primary care physician (PCP) visits per person over 6 months (vs the prior 6 months). Patient-reported outcomes included changes in patient activation, experience, health-related quality of life, and self-rated health at 6 months (vs baseline). We performed an intention-to-treat analysis using a linear mixed-effects model with a random intercept for each patient to examine the association between study group and outcomes. This study randomized 159 participants (mean [SD] age, 54.9 [9.8] years) to the A-ICU SUMMIT intervention (n = 80) or to EUC (n = 79). The majority of participants were men (102 [65.8%]) and most were White (121 [76.1%]). A total of 64 participants (41.0%) reported having unstable housing at baseline. Six-month hospitalizations decreased in both the A-ICU and EUC groups, with no difference between them (mean [SE], -0.6 [0.5] vs -0.9 [0.5]; difference, 0.3 [95% CI, -1.0 to 1.5]). Emergency department use did not differ between groups (mean [SE], -2.0 [1.0] vs 0.9 [1.0] visits per person; difference, -1.1 [95% CI, -3.7 to 1.6]). Primary care physician visits increased in the A-ICU group (mean [SE], 4.2 [1.6] vs -2.0 [1.6] per person; difference, 6.1 [95% CI, 1.8 to 10.4]). Patients in the A-ICU group reported improved social functioning (mean [SE], 4.7 [2.0] vs -1.1 [2.0]; difference, 5.8 [95% CI, 0.3 to 11.2]) and self-rated health (mean [SE], 0.7 [0.3] vs -0.2 [0.3]; difference, 1.0 [95% CI, 0.1 to 1.8]) compared with patients in the EUC group. No differences in patient activation or experience were observed. The A-ICU intervention did not change hospital or ED utilization at 6 months but increased PCP visits and improved patient well-being. Longer-term studies are needed to evaluate whether these observed improvements lead to eventual changes in acute care utilization. ClinicalTrials.gov Identifier: NCT03224858.


NADPH:Protochlorophyllide oxidoreductase uses the general import route into chloroplasts.
Chloroplast differentiation in angiosperm plants depends on the light-dependent conversion of protochlorophyllide to chlorophyllide by NADPH:protochlorophyllide oxidoreductase (PORA; EC 220.127.116.11), a nuclearly encoded protein. The protein import of the precursor form of PORA into plastids was shown previously to strictly depend on the presence of its substrate protochlorophyllide. PORA seemed to follow a novel, posttranslationally regulated import route. Here we demonstrate that the precursor of PORA from barley is imported into isolated barley plastids independently of protochlorophyllide. PORA as well as PORB import is competed for by the precursor of the small subunit of Rubisco. The data demonstrate that the PORA precursor uses the general import pathway into plastids. Furthermore, en route into chloroplasts the pea POR precursor can be cross-linked to the protein import channel in the outer envelope Toc75 from pea.


Pharmacologic activity of bepafant (WEB 2170), a new and selective hetrazepinoic antagonist of platelet activating factor.
The hetrazepine WEB 2170 (international nonproprietary name: bepafant), a thieno-triazolodiazepine that is structurally related to the recently described platelet activating factor (PAF) antagonist WEB 2086, is a potent and selective PAF antagonist both in vitro and in vivo. WEB 2170 inhibited PAF-induced human platelet and neutrophil aggregation in vitro (IC50 values: 0.3 and 0.83 microM, respectively) but had little or no inhibitory action against aggregation induced by other agonists. The potency in vitro was comparable to that described recently for WEB 2086 (Casals-Stenzel, J., Muacevic, G. and Weber, K.H.: J. Pharmacol. Exp. Ther. 241: 974-981, 1987). When guinea pigs were given i.v. infusions of PAF at 30 ng x kg-1 x min-1, oral (0.005-0.5 mg/kg) as well as intravenous (0.005-0.05 mg/kg) treatment with WEB 2170 abrogated the intrathoracic accumulation of 111In-labeled platelets, the bronchoconstriction and the hypotension as well as the finally occurring death in a dose-dependent fashion. Oral (0.01-1 mg/kg) and intravenous (0.005-0.1 mg/kg) WEB 2170 shared with the beta 2 agonist fenoterol and the steroid dexamethasone the property of protecting elderly NMRI mice from the lethal effect of PAF. In anesthetized rats, intravenous (0.001-0.1 mg/kg) and oral (0.05-1 mg/kg) WEB 2170 inhibited PAF-induced hypotension in a dose-related manner. Coadministration of WEB 2170 inhibited PAF-induced increase of vascular permeability in rat skin very effectively. The half-time of duration of action in the rat was estimated to be about 5 to 6 h after oral administration and about 1.1 to 2.3 h after intravenous application. In conclusion, the hetrazepine WEB 2170 is a strong and selective PAF antagonist, which is in vitro more or less equipotent to WEB 2086. In contrast, in vivo oral WEB 2170 is--depending on the species and considered parameter--about 5 to 40 times more potent against exogenous PAF-induced alterations than the recently described hetrazepine WEB 2086. Particularly in mice and rats, oral WEB 2170 is by far superior to WEB 2086.


Dynamic US contrast study of the liver: Vascular and delayed parenchymal phase.
We evaluated the time-intensity curves of the ultrasound contrast agent, Levovist, in the aorta, portal vein and liver parenchyma in order to define distribution of the agent when it is administered by an intravenous bolus injection. Twelve healthy volunteers were studied. All 12 subjects were examined for the study of vascular phase and five of the subjects were examined for the study of delayed parenchymal phase. To evaluate vascular enhancement, transverse abdominal scanning was performed. To evaluate parenchymal enhancement, liver scanning was done just once at 14 time points up to 60 min after injection. The time-intensity curves in the aorta and the portal vein indicated the conventional curves of blood pool agents such as iodine CT agents and gadolinium MRI agents. They showed steep initial rises and peaks at 20 s and 30 s after injection, respectively. Parenchymal enhancement reached a peak five minutes after injection, with a plateau for 20 min subsequently. It has been proved that there are two phases of Levovist contrast ultrasonography, the vascular phase and the delayed parenchymal phase.


Defining authorship and contributorship criteria: transparency and visibility.
The complexity and research publications have expanded exponentially.The role of authors and collaborators need to be clarified according with the standards of the International Committee of medical Journal Editors (ICMJE). New projects as Contributor Role Taxonomy (CRediT), has been launched in order to provide credit and transparency to the authors, readers and evaluation agencies.


Conservation of structure and subunit interactions in yeast homologues of splicing factor 3b (SF3b) subunits.
Human SAP 49, a subunit of the multimeric splicing factor 3b (SF3b), contains two RNA recognition motifs (RRMs) and binds another SF3b subunit called SAP 145, whose yeast homologue is CUS1. Here we show that the predicted yeast open reading frame YOR319w (HSH49) encodes an essential yeast splicing factor. Using bacterially expressed proteins, we find that yeast HSH49 binds CUS1. Mutations that alter putative RNA-binding residues of either HSH49 RRM are lethal in vivo, but do not prevent binding to CUS1 in vitro, suggesting that the predicted RNA-binding surfaces of HSH49 are not required for interaction with CUS1. In vivo interaction tests show that HSH49 and CUS1 associate primarily through the N-terminal RRM of HSH49. Recombinant HSH49 protein has a general RNA-binding activity that does not require CUS1. The parallels in structure and interaction between two SF3b subunits from yeast implies that the mechanism of SF3b action is highly conserved.


Intraluminal gel ultrasound and eco-color doppler: new tools for the study of colorectal cancer in mice.
Azoxymethane (AOM) is a potent carcinogen that induces colorectal cancer in mice. Intraluminal gel ultrasound is a technique based on the injection of gel into the rectum. This technique allows the colon to be straightened and to visualize and identify tumours. Twenty female C57Bl/6J mice were injected intraperitoneally with 10 mg/kg of AOM one time per week for six weeks. The mice were monitored by ultrasound with a Vevo 2100 system. We evaluated the tumour area and tumour vasculature with Ecocolor-Doppler (ECD). Histological examination of sacrificed mice was employed as the standard protocol. After 40 weeks from the injection, ultrasound analysis revealed the presence of tumours in 50% of all mice. Ex vivo analysis revealed the presence of 57% true-positives and only one false-positive. In two mice, ultrasound did not reveale the presence of tumour due to its small dimension. This indicates that ultrasound is able to detect only tumours with sizes ≥3 mm². Ultrasound is a rapid examination compared to other diagnostic techniques. It has a good sensitivity when the tumours reach the dimensions of 3 mm² or more. Intraluminal gel allows for the tumour area to be evaluated when mice are still alive, while ECD allows for vasculature of intestinal walls and colorectal tumour to be evaluated.


Characterization and role of suppressor of cytokine signaling 1a (SOCS1a) in a teleost fish, Miichthys miiuy.
The suppressor of cytokine signaling 1 (SOCS1) is a crucial regulator in the immune systems of mammals, which functions classically as a negatively regulator in the IFN signaling pathways. However, data on functional characterization of SOCS1 in lower vertebrates is limited. In this study, we identified and characterized the full-length SOCS1a gene of miiuy croaker (Miichthys miiuy). The sequence analysis results showed that miiuy croaker SOCS1a (mmSOCS1a) shared some conserved motifs with other vertebrates. To further study the function of fish SOCS1, we identified mmSOCS1a and determined its potential ability to perceive poly(I:C) stimulation. Induction experiments with poly(I:C) indicated the significant expression levels of mmSOCS1a in liver and kidney. In addition, mmSOCS1a could inhibit poly(I:C)-induced or IFNs-induced ISRE reporter gene activation. In a word, we systematically and comprehensively analyzed evolution and function of mmSOCS1a, which will provide the basis for future research on fish SOCS family.


[Demonstration of apolar lipids in paraffin-embedded tissues with special reference to pathological fatty degeneration in Tangier disease and other dyslipoproteinemias].
The oxidative cross-linking effect exerted by OsO4 on lipids and the blackening of osmiophilic structures resulting from reduction of OsO4 constitute long-known phenomena. Nevertheless, osmic acid has rarely been used for lipid staining as various even non-lipid-containing tissue structures are more or less intensively blackened by OsO4. If one treats osmicated tissue sections with certain oxidants (e.g. ammonium persulfate) in aqueous solution, then all the black OsO4 reduction products which are exposed to the aqueous phase are oxidized and thereby bleached. This applies e. g. to all proteins and polar (hydrophilic) lipids. Only apolar lipids retain their more or less black color and can be stained subsequently with Sudan dyes. Based on this principles, a new procedure has been developed which is effective on paraffin sections obtained after previous block osmication. This OBS procedure can also be used for very precise localization of appropriate lipid structures as can be demonstrated in cases of pathologic lipid deposits in an-alpha- and in a-beta-lipoproteinaemia, in type III hyperlipoproteinaemia, and in diverse other dyslipoproteinaemias as well.


Antimicrobial use of patients with sexually transmitted infection symptoms prior to presentation at five health facilities in Southern Ghana.
Unregulated and inappropriate antimicrobial use are major contributors to the evolution of antimicrobial resistance worldwide. It is important to monitor and collect data on the use of antibiotics at health facilities and in the general population in order to support antimicrobial stewardship programs. As part of a gonorrhea surveillance study that was conducted from June 2012 to Jan 2018, we administered a questionnaire to elicit information on the types of antimicrobials used by individuals to treat symptoms of a gonorrhea infection prior to presenting at five health facilities in Southern Ghana. Almost one-third (383/1,349; 28%) of study participants admitted taking one or more antimicrobial types before hospital presentation, while 138/383 (36%) of those who took antimicrobials could not remember what they ingested. A greater percentage of individuals who reported prior antimicrobial use before presentation at a health facility tested positive for gonorrhea by NAAT (30%), in contrast to 24% for those without prior treatment (p = 0.004). Penicillin and its derivatives, as well as ciprofloxacin and doxycycline, were the most used, while a few individuals reported taking drugs such as kanamycin and rifampin. Males were more likely than females to take an antimicrobial prior to attending a health center. In order to curb excessive and inappropriate antimicrobial use, antibiotics used by patients before presenting at hospitals ought to be investigated by healthcare providers. It is recommended that health professionals receive continuing education on the consequences of unregulated antimicrobial use.


The use and abuse of participatory action research.
This paper outlines the characteristics of PAR, participatory action research, so that interested investigators can determine if their project fits the criteria. The author introduces some key concepts in critical theory that drive the political analysis dimension of PAR and lay to rest concerns about validity and legitimacy. Types of PAR are reviewed according to the brand of knowledge focused on in the research, and some examples are given. An emerging critique of PAR is offered and some abuses of PAR are described. The major ethical principles of social justice that guide PAR are outlined. References to epistemology and specific methodological demonstration projects are included. Suggestions for future use in chronic disease research are offered.


Sevoflurane induces microRNA-18a to delay rat neurodevelopment via suppression of the RUNX1/Wnt/β-catenin axis.
Sevoflurane anesthesia is reported to repress neurogenesis of neural stem cells (NSCs), thereby affecting the brain development, but the underlying mechanism of sevoflurane on the proliferation of NSCs remains unclear. Thus, this study aims to discern the relationship between sevoflurane and NSC proliferation. Bioinformatics tools were employed to predict the expression of microRNA-18a (miR-18a) in 9-day-old neonatal rat hippocampal tissues after sevoflurane treatment and the downstream genes of miR-18a, followed by a series of assays to explore the relationship among miR-18a, runt related transcription factor 1 (RUNX1), and β-catenin in the hippocampal tissues. NSCs were isolated from the hippocampal tissues and subjected to gain-/loss-of-function assays to investigate the interactions among miR-18a, RUNX1, and β-catenin in NSCs and their roles in NSC development. Bioinformatics analysis and experimental results confirmed high expression of miR-18a in rat hippocampal tissues and NSCs after sevoflurane treatment. Next, we found that miR-18a downregulated RUNX1 expression, while RUNX1 promoted NSC proliferation by activating the Wnt/β-catenin signaling pathway. The behavioral experiments also showed that sevoflurane caused nerve injury in rats, whilst RUNX1 overexpression protected rat neurodevelopment. Our findings uncovered that sevoflurane attenuated NSC proliferation via the miR-18a-meidated RUNX1/Wnt/β-catenin pathway, thereby impairing rat neurodevelopment.


[The treatment of urolithiasis in the specialized urologic department].
The experience of urolithiasis treatment with the use of modern diagnostic methods (the computer tomography with concrement density detection, the DV-Med program which permits to obtain the calculus volumetric image and to study its physical characteristics), and therapy (distant lithotripsy with the use of different types of lithotriptors as well as retrograde contact lithotripsy with "Lithoclast") permits to minimize the percent of open surgical interventions. For the last 3 years 780 patients with different clinical disease forms were treated with favorable clinical outcome and only in 0.5% cases the open surgical intervention was conducted. These patients underwent the pyelolithotomy for the secondary nephrolithiasis conditioned by the pelvis-and-ureteral segment stricture when the endoscopic method failed to eliminate this problem.


Diphenyleneiodonium sulfate, an NADPH oxidase inhibitor, prevents early alcohol-induced liver injury in the rat.
The oxidant source in alcohol-induced liver disease remains unclear. NADPH oxidase (mainly in liver Kupffer cells and infiltrating neutrophils) could be a potential free radical source. We aimed to determine if NADPH oxidase inhibitor diphenyleneiodonium sulfate (DPI) affects nuclear factor-kappaB (NF-kappaB) activation, liver tumor necrosis factor-alpha (TNF-alpha) mRNA expression, and early alcohol-induced liver injury in rats. Male Wistar rats were fed high-fat liquid diets with or without ethanol (10-16 g. kg(-1). day(-1)) continuously for up to 4 wk, using the Tsukamoto-French intragastric enteral feeding protocol. DPI or saline vehicle was administered by subcutaneous injection for 4 wk. Mean urine ethanol concentrations were similar between the ethanol- and ethanol plus DPI-treated groups. Enteral ethanol feeding caused severe fat accumulation, mild inflammation, and necrosis in the liver (pathology score, 4.3 +/- 0.3). In contrast, DPI significantly blunted these changes (pathology score, 0.8 +/- 0.4). Enteral ethanol administration for 4 wk also significantly increased free radical adduct formation, NF-kappaB activity, and TNF-alpha expression in the liver. DPI almost completely blunted these parameters. These results indicate that DPI prevents early alcohol-induced liver injury, most likely by inhibiting free radical formation via NADPH oxidase, thereby preventing NF-kappaB activation and TNF-alpha mRNA expression in the liver.


Colposcopically observed vascular changes in the cervix in relation to the hormonal levels and menstrual cycle.
We assessed colposcopically observed vascular changes occurring in the cervix in relation to cyclical hormonal variation in healthy women. Thirty women with regular menstrual cycles and willing to remain sexually abstinent during a menstrual cycle were enrolled. Colposcopy was performed during the peak of the estrogen and progesterone levels. The mean (+/-SD) diameter of the largest visible blood vessel differed significantly between the estrogenic phase (0.38 +/- 0.14 mm) as compared with the progestogenic phase (0.47 +/- 0.12 mm; p <.01). The blood vessels were more prominent and dense and had a well-defined outline during the progestogenic phase than the estrogenic phase; however, these differences were not statistically significant. There was borderline increase in the interleukin 8 level during the estrogenic phase. Physiological changes of increased vascularity of the cervix observed colposcopically during the progestogenic phase are normal. If such changes do not correspond to the menstrual cycle phase in women using vaginal microbicides in early-phase clinical trials, presence of inflammatory markers should be evaluated. Elevated interleukin 8 during the estrogenic phase needs further evaluation.


Efficacy and limitations of ChatGPT as a biostatistical problem-solving tool in medical education in Serbia: a descriptive study
This study aimed to assess the performance of ChatGPT (GPT-3.5 and GPT-4) as a study tool in solving biostatistical problems and to identify any potential drawbacks that might arise from using ChatGPT in medical education, particularly in solving practical biostatistical problems. ChatGPT was tested to evaluate its ability to solve biostatistical problems from the Handbook of Medical Statistics by Peacock and Peacock in this descriptive study. Tables from the problems were transformed into textual questions. Ten biostatistical problems were randomly chosen and used as text-based input for conversation with ChatGPT (versions 3.5 and 4). GPT-3.5 solved 5 practical problems in the first attempt, related to categorical data, cross-sectional study, measuring reliability, probability properties, and the t-test. GPT-3.5 failed to provide correct answers regarding analysis of variance, the chi-square test, and sample size within 3 attempts. GPT-4 also solved a task related to the confidence interval in the first attempt and solved all questions within 3 attempts, with precise guidance and monitoring. The assessment of both versions of ChatGPT performance in 10 biostatistical problems revealed that GPT-3.5 and 4's performance was below average, with correct response rates of 5 and 6 out of 10 on the first attempt. GPT-4 succeeded in providing all correct answers within 3 attempts. These findings indicate that students must be aware that this tool, even when providing and calculating different statistical analyses, can be wrong, and they should be aware of ChatGPT's limitations and be careful when incorporating this model into medical education.


Hysteria and conversion in the ojibway patient: cross-cultural psychiatry for the family physician.
Case examples of hysterical coma and paralysis in two Ojibway patients are used to introduce a discussion of conversion symptoms. Somatoform disorders are described using the criteria of the Diagnostic and Statistical Manual of Mental Disorders, third edition; these are then applied to hysteria in the Native American patient. The authors discuss the cultural context of conversion symptoms in this population and describe guidelines for the provision of culturally sensitive therapy by the family physician.


Production of sterile mono-sex triploid yellow drum (Nibea albiflora): genotypic females and sex-reversed phenotypic males with emphasis on utilization as surrogate broodstock.
Production of sterile mono-sex fish is of great significance for sustainable aquaculture as well as germ cell transplantation. In this study, we aimed to produce mono-sex triploid yellow drum, including genotypic females (XXX female) and sex-reversed phenotypic males (XXX male). Firstly, the mono-female triploids were produced through cold-shock treatment on eggs fertilized with sperm from neo-males. Then, the mono-male triploids were produced by the sex reversal of mono-female triploids with oral administration of letrozole (LZ). We comparatively investigated the growth and gonadal development in the mono-sex triploids. The results showed that the triploids displayed similar growth performance to their diploids throughout their first year, but had impaired gonadosomatic index and gametogenesis. No mature gametes were produced in the triploids during their first spawning season. Meanwhile, we analyzed the process of gametogenesis in the both sex of triploids. Ultrastructure of gametogenesis showed that the germ cells arrested at abnormal metaphase 1 in females, while males had irregular meiotic divisions, variable-sized spermatid and degenerated cells. The expression levels of meiosis-related genes (i.e., sycp3 and rec8) confirmed the abnormal meiosis in the triploids. Furthermore, the gonadal development was also determined by the expression patterns of vasa, dmrt1 and cyp19a1a. Abnormal expression of vasa mRNA and protein were detected in triploids. High cyp19a1a expression levels suggested the sex steroid hormones production might be at least partially functional in triploid females. In addition, high dmrt1 expression levels confirmed the masculinization and testicular development of sex-reversed triploid males by LZ. Our findings provide an efficient protocol to produce sterile mono-sex triploid yellow drum and provide new insights into the mechanism of gonadal sterility of triploid fish.


Cytotoxic constituents from the mangrove endophytic Pestalotiopsis sp. induce G0/G1 cell cycle arrest and apoptosis in human cancer cells.
Chemical examination of Chinese mangrove Rhizophora mucronata endophytic Pestalotiopsis sp., yielded 11 known metabolites with various structure types, including demethylincisterol A3 (1), dankasterone B (2), (22E, 24R)-ergosta-7,9(11), 22-triene-3β, 5α, 6α-triol (3), ergosta-5,7,22-trien-3-ol (4), 5, 8-epidioxy-5, 8-ergosta-6, 22E-dien-3-ol (5), stigmastan-3-one (6), stigmast-4-en-3-one (7), stigmast-4-en-6 -ol-3-one (8), flufuran (9), (2-cis, 4-trans)-abscisic acid (10), similanpyrone B (11). Their structures were unambiguously elucidated on the basis of extensive NMR spectroscopic and mass spectrometric analyses. Compounds 1, 4, 6-9 showed significant in vitro cytotoxicity against the human cancer cell lines Hela, A549 and HepG, of which compound 1 was the most potential with IC50 values reaching nM degree ranging from 0.17 to 14.16 nM. Flow cytometric investigation demonstrated that compound 1 mainly inhibited cell cycle at G0/G1 phase in a dose-dependent manner with a significant induction of apoptosis on the three tested cell lines. The involvement of the mitochondria in compound 1 induced apoptosis was investigated using MMP. We suggested that R. mucronata endophytic Pestalotiopsis sp. contained a potential anticancer compound demethylincisterol A3.


15-Deoxy-Delta12,1412,14-prostaglandin J2 inhibits the beta2 integrin-dependent oxidative burst: involvement of a mechanism distinct from peroxisome proliferator-activated receptor gamma ligation.
15-Deoxy-Delta12,14-PGJ2 (dPGJ2) is a bioactive metabolite of the J2 series that has been identified as a ligand for peroxisome proliferator-activated receptor gamma (PPARgamma) and has received attention for its potential antiinflammatory effects. Because neutrophils express cell-surface receptors for PGs, the effect of dPGJ2 was tested on an inflammatory response that should not require PPARgamma, the oxidative burst made by adherent human neutrophils. dPGJ2 inhibited adhesion-dependent H2O2 production with an IC50 of 1. 5 microM when neutrophils were stimulated with TNF, N-formylnorleucylleucylphenylalanine, or LPS. Inhibition by dPGJ2 occurred during the lag phase, before generation of peroxide, suggesting blockade of an early signaling step. Indeed, dPGJ2 blocked adhesion of neutrophils to fibrinogen in response to TNF or LPS with an IC50 of 3-5 micro+dPGJ2 was more potent at inhibiting the adhesion-dependent oxidative burst than several other PGs tested. Further, dPGJ2 did not appear to act through either the DP receptor or receptors for PGE2. PG receptors modulate cAMP levels, and the inhibition of adhesion and oxidative burst by dPGJ2 was enhanced in the presence of 3-isobutyl-1-methylxanthine, a cAMP phosphodiesterase inhibitor. A potent PPARgamma agonist (AD-5075) did not inhibit peroxide production or adhesion, nor did it change the IC50 for dPGJ2 inhibition. These studies suggest that dPGJ2 may interact with an unknown receptor on neutrophils, distinct from PPARgamma, to modulate the production of reactive oxygen intermediates.


Prevalence of fatigue and its association with clinical features in progressive and non-progressive forms of Multiple Sclerosis.
Fatigue is a complex and disabling symptom of Multiple Sclerosis (MS); however, there is conflicting evidence of the relationship between fatigue and clinical features of MS. Furthermore, few studies have considered these relationships specifically in a progressive MS population. (1) estimate the prevalence of self-reported fatigue in people with MS; (2) evaluate the relationship between fatigue severity/impact and clinical features of MS; (3) compare the prevalence of fatigue, and the strength of relationship between fatigue severity/impact and clinical features of MS in progressive and non-progressive forms of MS. An online survey was conducted to measure the severity (Fatigue Severity Scale (FSS)) and impact of self-reported fatigue (Modified Fatigue Impact Scale) in people with MS. The survey also contained questionnaires related to disability, quality of life, MS impact, anxiety and depression, cognition, and sleep quality. 412 people responded to the survey, of which 68.7% reported having fatigue (FSS ≥ 5). The prevalence of fatigue was significantly higher in participants with progressive MS (81%) in comparison to those with non-progressive forms of MS (64%, p = 0.01). Fatigue severity and impact were associated with quality of life, MS impact, anxiety, depression, cognition, and sleep quality in both progressive and non-progressive MS populations (p < 0.05). However, fatigue severity (r = 0.335) and impact (r = 0.391) were correlated with disability only in participants with non-progressive MS. Fatigue was more prevalent amongst participants with progressive MS. In addition, higher fatigue severity and impact were associated with greater physical, cognitive, and psychological impairment, although the strength of association between these outcomes was generally similar regardless of the type of MS.


The burden of hospitalised carcinoma patients in Osijek-Baranja County, Croatia, 1998-2010.
The purpose of this paper was to determine the number and trend of hospitalisations caused by carcinoma in the Osijek-Baranja County, to to determine the proportion of these hospitalizations compared to all other hospitalization, as well as to determine the burden put on the hospitals by external patients (those not residing in the Osijek-Baranja County area). There has been an average of 10.1% of hospitalisations caused by carcinoma in the Osijek-Baranja County from 1998 to 2010. In the aforementioned period there have been an average 15.9% days of hospitalisation caused by carcinoma. Patients hospitalised due to carcinoma stay in hospital 5.3 days longer than other patients. The number of hospitalisations as a whole, as well as those due to carcinoma, is rising, while the number of days of hospitalisation per patient is decreasing. The number of hospitalisations which refer to men is slightly higher than for women (51.2%:48.8%). Almost one third of patients hospitalised are not residents of the Osijek-Baranja County due to a gravitational hospitalisation instance. Residents of northern Bosnia and Herzegovina are the most common patients being treated in our county. This burden has to be taken in to account when planning our health care due to the geographically, economically and politically specific situation of Osijek-Baranya County. This data has to be taken in to account when further planning our health care, as to relieve the acute medical situation hospitals of some of their burden, as well as to provide adequate care to patients suffering chronically from carcinoma. It is especially important to stress out that our county does not have a palliative care facility, nor a hospice for the terminally ill.


A Computational Model of Thalamocortical Dysrhythmia in People With Tinnitus.
Tinnitus is a problem that affects a diverse range of people. One common trait amongst people with tinnitus is the presence of hearing loss, which is apparent in over 90% of the cohort. It is postulated that the remainder of people with tinnitus have hidden hearing loss in the form of cochlear synaptopathy. The loss of hearing sensation is thought to cause a reduction in the bottom-up excitatory signals of the auditory pathway leading to a change in the frequency of thalamocortical oscillations known as thalamocortical dysrhythmia (TCD). The downward shift in oscillatory behavior, characteristic of TCD, has been recorded experimentally but the underlying mechanisms responsible for TCD in tinnitus subjects cannot be directly observed. This paper investigates these underlying mechanisms by creating a biologically faithful model of the auditory periphery and thalamocortical network, called the central auditory processing (CAP) model. The proposed model replicates tinnitus related activity in the presence of hearing loss and hidden hearing loss in the form of cochlear synaptopathy. The results of this paper show that, both the bottom-up and top-down changes are required in the auditory system for tinnitus related hyperactivity to coexist with TCD, contrary to the theoretical model for TCD. The CAP model provides a novel modeling approach to account for tinnitus related activity with and without hearing loss. Moreover, the results provide additional clarity to the understanding of TCD and tinnitus and provide direction for future approaches to treating tinnitus.


Action potential duration heterogeneity of cardiac tissue can be evaluated from cell properties using Gaussian Green's function approach.
Action potential duration (APD) heterogeneity of cardiac tissue is one of the most important factors underlying initiation of deadly cardiac arrhythmias. In many cases such heterogeneity can be measured at tissue level only, while it originates from differences between the individual cardiac cells. The extent of heterogeneity at tissue and single cell level can differ substantially and in many cases it is important to know the relation between them. Here we study effects from cell coupling on APD heterogeneity in cardiac tissue in numerical simulations using the ionic TP06 model for human cardiac tissue. We show that the effect of cell coupling on APD heterogeneity can be described mathematically using a Gaussian Green's function approach. This relates the problem of electrotonic interactions to a wide range of classical problems in physics, chemistry and biology, for which robust methods exist. We show that, both for determining effects of tissue heterogeneity from cell heterogeneity (forward problem) as well as for determining cell properties from tissue level measurements (inverse problem), this approach is promising. We illustrate the solution of the forward and inverse problem on several examples of 1D and 2D systems.


Diabetes and hypertension: public awareness and lifestyle - findings of a health mela.
To determine the frequencies of diabetes, hypertension and their established lifestyle risk factors and to assess the level of awareness about diabetes and hypertension amongst persons attending a health mela at the Aga Khan University Hospital (AKUH), Karachi. Cross-sectional, analytical study. One-day health mela held at AKUH, Karachi. A total of 264 participants were administered structured questionnaire to obtain demographic data and perceptions about diabetes and hypertension after taking verbal informed consent. Height, weight, blood pressure and random blood glucose were measured. Overall frequency of type 2 diabetes was 13.6%, impaired glucose tolerance (IGT) 8.3% and hypertension 24%. Frequency of diabetes and hypertension in both men and women increased with increasing age(p<0.001) and body mass index (p=0.02). Over half the men and women with type 2 diabetes (53% and 57% respectively) and 42% men and 60% women with IGT also had hypertension. Diabetes and hypertension were correctly defined by 52% and 37% subjects respectively and this was significantly associated with educational level (p=.001). Lack of physical exercise was observed in 59% participants, while 53.6% men and 67.5% women were overweight/obese. As compared to women, men used more additional salt (p=0.03) and had more outside meals (p<0.001) and snacks (p=0.01). High frequencies of diabetes, hypertension, obesity, unhealthy nutrition and lack of exercise were observed in the study population. Emphasis on health education is needed to increase public awareness of the warning signs and risk factors of these common conditions.


Human Action Recognition in Unconstrained Videos by Explicit Motion Modeling.
Human action recognition in unconstrained videos is a challenging problem with many applications. Most state-of-the-art approaches adopted the well-known bag-of-features representations, generated based on isolated local patches or patch trajectories, where motion patterns, such as object-object and object-background relationships are mostly discarded. In this paper, we propose a simple representation aiming at modeling these motion relationships. We adopt global and local reference points to explicitly characterize motion information, so that the final representation is more robust to camera movements, which widely exist in unconstrained videos. Our approach operates on the top of visual codewords generated on dense local patch trajectories, and therefore, does not require foreground-background separation, which is normally a critical and difficult step in modeling object relationships. Through an extensive set of experimental evaluations, we show that the proposed representation produces a very competitive performance on several challenging benchmark data sets. Further combining it with the standard bag-of-features or Fisher vector representations can lead to substantial improvements.


BDNF protects the neonatal brain from hypoxic-ischemic injury in vivo via the ERK pathway.
Neurotrophins activate several different intracellular signaling pathways that in some way exert neuroprotective effects. In vitro studies of sympathetic and cerebellar granule neurons have demonstrated that the survival effects of neurotrophins can be mediated via activation of the phosphatidylinositol 3-kinase (PI3-kinase) pathway. Neurotrophin-mediated protection of other neuronal types in vitro can be mediated via the extracellular signal-related protein kinase (ERK) pathway. Whether either of these pathways contributes to the neuroprotective effects of neurotrophins in the brain in vivo has not been determined. Brain-derived neurotrophic factor (BDNF) is markedly neuroprotective against neonatal hypoxic-ischemic (H-I) brain injury in vivo. We assessed the role of the ERK and PI3-kinase pathways in neonatal H-I brain injury in the presence and absence of BDNF. Intracerebroventricular administration of BDNF to postnatal day 7 rats resulted in phosphorylation of ERK1/2 and the PI3-kinase substrate AKT within minutes. Effects were greater on ERK activation and occurred in neurons. Pharmacological inhibition of ERK, but not the PI3-kinase pathway, inhibited the ability of BDNF to block H-I-induced caspase-3 activation and tissue loss. These findings suggest that neuronal ERK activation in the neonatal brain mediates neuroprotection against H-I brain injury, a model of cerebral palsy.


A flexible, redox-active macrocycle enables the electrocatalytic reduction of nitrate to ammonia by a cobalt complex.
The cobalt macrocycle complex [Co(DIM)Br2]+ (DIM = 2,3-dimethyl-1,4,8,11-tetraazacyclotetradeca-1,3-diene) is an electrocatalyst for the selective reduction of nitrate to ammonia in aqueous solution. The catalyst operates over a wide pH range and with very high faradaic efficiency, albeit with large overpotential. Experimental investigations, supported by electronic structure calculations, reveal that catalysis commences when nitrate binds to the two-electron reduced species CoII(DIM-), where cobalt and the macrocycle are each reduced by a single electron. Several mechanisms for the initial reduction of nitrate to nitrite were explored computationally and found to be feasible at room temperature. The reduced DIM ligand plays an important role in these mechanisms by directly transferring a single electron to the bound nitrate substrate, activating it for further reactions. These studies further reveal that the DIM macrocycle is critical to nitrate reduction, specifically its combination of redox non-innocence, hydrogen-bonding functionality and flexibility in coordination mode.


Virus-specific CD8+ T cells upregulate programmed death-1 expression during acute friend retrovirus infection but are highly cytotoxic and control virus replication.
It was recently reported that inhibitory molecules such as programmed death-1 (PD-1) were upregulated on CD8(+) T cells during acute Friend retrovirus infection and that the cells were prematurely exhausted and dysfunctional in vitro. The current study confirms that most activated CD8(+) T cells upregulated expression of PD-1 during acute infection and revealed a dichotomy of function between PD-1(hi) and PD-1(lo) subsets. More PD-1(lo) cells produced antiviral cytokines such as IFN-γ and TNF-α, whereas more PD-1(hi) cells displayed characteristics of cytotoxic effectors such as production of granzymes and surface expression of CD107a. Importantly, CD8(+) T cells mediated rapid in vivo cytotoxicity and were critical for control of acute Friend virus replication. Thus, direct ex vivo analyses and in vivo experiments revealed high CD8(+) T cell functionality and indicate that PD-1 expression during acute infection is not a marker of T cell exhaustion.


Medication supply chain management through implementation of a hospital pharmacy computerized inventory program in Haiti.
In the aftermath of the 2010 earthquake in Haiti, St. Luke Hospital was built to help manage the mass casualties and subsequent cholera epidemic. A major problem faced by the hospital system was the lack of an available and sustainable supply of medications. Long-term viability of the hospital system depended largely on developing an uninterrupted medication supply chain. We hypothesized that the implementation of a new Pharmacy Computerized Inventory Program (PCIP) would optimize medication availability and decrease medication shortages. We conducted the research by examining how medications were being utilized and distributed before and after the implementation of PCIP. We measured the number of documented medication transactions in both Phase 1 and Phase 2 as well as user logins to determine if a computerized inventory system would be beneficial in providing a sustainable, long-term solution to their medication management needs. The PCIP incorporated drug ordering, filling the drug requests, distribution, and dispensing of the medications in multiple settings; inventory of currently shelved medications; and graphic reporting of 'real-time' medication usage. During the PCIP initiation and establishment periods, the number of medication transactions increased from 219.6 to 359.5 (p=0.055), respectively, and the mean logins per day increased from 24.3 to 31.5, p<0.0001, respectively. The PCIP allows the hospital staff to identify and order medications with a critically low supply as well as track usage for future medication needs. The pharmacy and nursing staff found the PCIP to be efficient and a significant improvement in their medication utilization. An efficient, customizable, and cost-sensitive PCIP can improve drug inventory management in a simplified and sustainable manner within a resource-constrained hospital.


Multivariate pattern analysis utilizing structural or functional MRI-In individuals with musculoskeletal pain and healthy controls: A systematic review.
The purpose of this systematic review is to systematically review the evidence relating to findings generated by multivariate pattern analysis (MVPA) following structural or functional magnetic resonance imaging (fMRI) to determine if this analysis is able to: a) Discriminate between individuals with musculoskeletal pain and healthy controls, b) Predict pain perception in healthy individuals stimulated with a noxious stimulus compared to those stimulated with a non-noxious stimulus. MEDLINE, CINAHL, Embase, PEDro, Google Scholar, Cochrane library and Web of Science were systematically screened for relevant literature using different combinations of keywords regarding structural and functional MRI analysed with MVPA, both in individuals with musculoskeletal pain and healthy controls. Reference lists of included articles were hand-searched for additional literature. Eligible articles were assessed on risk of bias and reviewed by two independent researchers. The search query returned 18 articles meeting the inclusion criteria. Methodological quality varied from poor to good. Seven studies investigated the ability of machine-learning algorithms to differentiate patient groups from healthy control participants. Overall, the review demonstrated that MVPA can discriminate between individuals with MSK pain and healthy controls with an overall accuracy ranging from 53% to 94%. Twelve studies utilized healthy control participants (using them as their own controls), during experimental pain paradigms aimed to investigate the ability of machine-learning to differentiate individuals stimulated with noxious stimuli from those stimulated with non-noxious stimuli, with 'pain' detection rates ranging from 60% to 94%. However, significant heterogeneity in patient conditions, study methodology and brain imaging techniques resulted in various findings that make study comparisons and formal conclusions challenging. There is preliminary and emerging evidence that MVPA analyses of structural or functional MRI are able to discriminate between patients and healthy controls, and also discriminate between noxious and non-noxious stimulation. No prospective studies were found in this review to allow determination of the prognostic or diagnostic capabilities or treatment responsiveness of these analyses. Future studies would also benefit from combining various behavioural, genotype and phenotype data into analyses to assist with development of sensitive and specific signatures that could guide future individualized patient treatment options and evaluate how treatments exert their effects.


Biomechanical estimation of tennis serve using inertial sensors: A case study.
Inertial measurement units may provide a relevant on-court 3-Dimension measurement system for tennis serve biomechanical analysis. Therefore, this case study aimed to report the feasibility of inertial measurement unit's kinematic and kinetic data collection during tennis serve. Two injury-free highly-trained tennis players were equipped with the inertial measurement unit (Xsens MVN suit) and performed 2 trials of five flat "first" serves on a 1 m2 target zone bordering the service box of an indoor GreenSet® tennis court surface. With the exception of the center of gravity rotation at the loading stage, all joint (shoulder, elbow, knee) angles, center of mass displacements and rotations followed a similar development for both female and male participants from loading to finish stages. At ball contact stage, articular moments (mid-trunk, upper-trunk, shoulder, elbow, wrist) and segmental contribution (pelvis linear, pelvis rotation, trunk, shoulder, elbow, wrist) repartitions also showed a comparable movement. From loading to finish stages, total, lower and upper energy contribution were similar for both players, with coefficient of variations deemed acceptable between the two trials. This inertial measurement unit appears suitable for on-court tennis serve biomechanical data collection and subsequent analysis to provide tennis players and practitioners tailored feedbacks to facilitate motor learning process and develop serve efficiency.


[Comparative assessment of electroencephalographic readings and perfusion index in patients in the perioperative period and coma].
Different monitoring modes are used to estimate the depth of anesthesia. Electroencephalography (EEG) is employed as a method of an electrophysiological study of brain function both during anesthesia and at the neuroresuscitative stage and changes in the peripheral vascular bed are evaluated by means of the perfusion index (PI). The purpose of the study was to comparatively analyze the use of EEG readings and PI at different stages of surgery and in coma of various etiologies. Thirty female patients aged 30-70 years (Group 1) operated on under general anesthesia were examined. Group 2 consisted of 14 intensive care unit patients of both sexes and different ages (range, 3 to 60 years) who had depressed consciousness to the comatose level. The association established between the changes in EEG reading and PI in the perioperative period allows estimation of the depth and adequacy of anesthesia, by analyzing the changes in both EEG and PI. Unlike central hemodynamic parameters, PI shows an earlier response to nociception. Group 2 patients had also synchronous changes in EEG and PI characteristics in various clinical conditions.


Hyperpolarized [1-13C] glutamate: a metabolic imaging biomarker of IDH1 mutational status in glioma.
Mutations of the isocitrate dehydrogenase 1 (IDH1) gene are among the most prevalent in low-grade glioma and secondary glioblastoma, represent an early pathogenic event, and are associated with epigenetically driven modulations of metabolism. Of particular interest is the recently uncovered relationship between the IDH1 mutation and decreased activity of the branched-chain amino acid transaminase 1 (BCAT1) enzyme. Noninvasive imaging methods that can assess BCAT1 activity could therefore improve detection of mutant IDH1 tumors and aid in developing and monitoring new targeted therapies. BCAT1 catalyzes the transamination of branched-chain amino acids while converting α-ketoglutarate (α-KG) to glutamate. Our goal was to use (13)C magnetic resonance spectroscopy to probe the conversion of hyperpolarized [1-(13)C] α-KG to hyperpolarized [1-(13)C] glutamate as a readout of BCAT1 activity. We investigated two isogenic glioblastoma lines that differed only in their IDH1 status and performed experiments in live cells and in vivo in rat orthotopic tumors. Following injection of hyperpolarized [1-(13)C] α-KG, hyperpolarized [1-(13)C] glutamate production was detected both in cells and in vivo, and the level of hyperpolarized [1-(13)C] glutamate was significantly lower in mutant IDH1 cells and tumors compared with their IDH1-wild-type counterparts. Importantly however, in our cells the observed drop in hyperpolarized [1-(13)C] glutamate was likely mediated not only by a drop in BCAT1 activity, but also by reductions in aspartate transaminase and glutamate dehydrogenase activities, suggesting additional metabolic reprogramming at least in our model. Hyperpolarized [1-(13)C] glutamate could thus inform on multiple mutant IDH1-associated metabolic events that mediate reduced glutamate production.


Dimension reduction with gene expression data using targeted variable importance measurement.
When a large number of candidate variables are present, a dimension reduction procedure is usually conducted to reduce the variable space before the subsequent analysis is carried out. The goal of dimension reduction is to find a list of candidate genes with a more operable length ideally including all the relevant genes. Leaving many uninformative genes in the analysis can lead to biased estimates and reduced power. Therefore, dimension reduction is often considered a necessary predecessor of the analysis because it can not only reduce the cost of handling numerous variables, but also has the potential to improve the performance of the downstream analysis algorithms. We propose a TMLE-VIM dimension reduction procedure based on the variable importance measurement (VIM) in the frame work of targeted maximum likelihood estimation (TMLE). TMLE is an extension of maximum likelihood estimation targeting the parameter of interest. TMLE-VIM is a two-stage procedure. The first stage resorts to a machine learning algorithm, and the second step improves the first stage estimation with respect to the parameter of interest. We demonstrate with simulations and data analyses that our approach not only enjoys the prediction power of machine learning algorithms, but also accounts for the correlation structures among variables and therefore produces better variable rankings. When utilized in dimension reduction, TMLE-VIM can help to obtain the shortest possible list with the most truly associated variables.


Successful multimodal treatment of a breast cancer patient with a recurrence invading the chest wall.
We describe successful operative management of a solitary breast cancer metastasis in the chest wall after complete response with concomitant non-pegylated liposomal doxorubicin (NPLD) and docetaxel followed by sternal rib resection with prosthetic reconstruction. We report a case of a 41-year-old woman who had a breast cancer recurrence infiltrating neighboring osteo-cartilage of the left sternal body, the cartilaginous portion of the third and fourth ipsilateral ribs and was inseparable from the rear side pectoral reaching deep into contiguity with the pericardium. After 6 cycles of chemotherapy with NPLD plus docetaxel, sternal rib resection with prosthetic reconstruction was performed. Histological examination did not show any evidence of residual tumor. At 9 months of follow-up, the patient appears free of disease. Our case demonstrates that a multimodal approach in patients with chest wall recurrence of breast cancer without distant metastasis, may be safe and effective for maintaining a good quality of life.


An enzymatic deglycosylation scheme enabling identification of core fucosylated N-glycans and O-glycosylation site mapping of human plasma proteins.
Global proteome analysis of protein glycosylation is a major challenge due to the inherent heterogeneous and diverse nature of this post-translational modification. It is therefore common to enzymatically remove glycans attached to protein or peptide chains prior to mass spectrometric analysis, thereby reducing the complexity and facilitating glycosylation site determinations. Here, we have used two different enzymatic deglycosylation strategies for N-glycosylation site analysis. (1) Removal of entire N-glycan chains by peptide-N-glycosidase (PNGase) digestion, with concomitant deamidation of the released asparagine residue. The reaction is carried out in H218O to facilitate identification of the formerly glycosylated peptide by incorporatation of 18O into the formed aspartic acid residue. (2) Digestion with two endo-beta-N-acetylglucosaminidases (Endo D and Endo H) that cleave the glycosidic bond between the two N-acetylglucosamine (GlcNAc) residues in the conserved N-glycan core structure, leaving single GlcNAc residues with putative fucosyl side chains attached to the peptide. To enable digestion of complex and hybrid type N-glycans, a number of exoglycosidases (beta-galactosidase, neuraminidase and N-acetyl-beta-glucosaminidase) are also included. The two strategies were here applied to identify 103 N-glycosylation sites in the Cohn IV fraction of human plasma. In addition, Endo D/H digestion uniquely enabled identification of 23 fucosylated N-glycosylation sites. Several O-glycosylated peptides were also identified with a single N-acetylhexosamine attached, arguably due to partial deglycosylation of O-glycan structures by the exoglycosidases used together with Endo D/H.


Effectiveness of ten-valent pneumococcal conjugate vaccine on invasive pneumococcal disease among children <2 years old: A prospective population-based study in rural Bangladesh.
The 10-valent pneumococcal conjugate vaccine (PCV10) was introduced in March 2015 in Bangladesh. In this study, we aimed to estimate the impact of PCV10 on invasive pneumococcal disease (IPD) identified by blood cultures and severe pneumonia identified clinically and its effectiveness on invasive disease caused by vaccine serotypes. We conducted population-based surveillance among children aged 2- <24 months between April 2012 through March 2019 in Mirzapur, a rural sub-district of Bangladesh. We compared incidence of IPD and severe pneumonia before (April 2012 to March 2015) and after (April 2015 to March 2019) the introduction of PCV10. Vaccine effectiveness was measured using an indirect cohort analysis of data from four sentinel sites in which PCV10 vaccination status was compared between children with IPD caused by vaccine serotype vs. non-vaccine serotypes. We identified 24 IPD cases by blood culture and 1,704 severe pneumonia hospitalizations during the surveillance period. IPD incidence in under-2-year-old children fell 25 % (95 % CI: -1.2 % to 76 %; p-value = 0.59) from 106 cases per 100,000 child-years at baseline to 79.3 in April 2018- March 2019. Vaccine serotype-IPD incidence was lower (77 % reduction, 95 % CI: -0.45 % to 96 %; p-value = 0.068) in April 2018 - March 2019 than in the pre-vaccine period (85.7 cases to 19.8/100,000 child-years). A significant decline of 54.0 % (95 % CI: 47.0 % to 59.0 %; p-value < 0.001) was observed in hospitalizations due to severe pneumonia. From indirect cohort analysis, the effectiveness of PCV10 against vaccine serotype IPD was 37 % (95 % CI: -141.0 % to 83.5 %; p = 0.5) after the 1st dose and 63.1 % (95 % CI: -3.3 % to 85.9 %, p = 0.0411) after the 2nd or the 3rd dose. This study demonstrates that PCV10 introduction prevented hospitalizations with severe pneumonia and provided individual protection against vaccine serotypes.


Virtual Monochromatic Dual-Energy Aortoiliac CT Angiography With Reduced Iodine Dose: A Prospective Randomized Study.
The purpose of this study was to assess the feasibility of performing abdominopelvic aortoiliac CT angiography (CTA) with 16.0 g of iodine contrast medium acquired with low-energy (40 and 50 keV) virtual monochromatic (VMC) images with rapid-kilovoltage-switching dual-energy CT. A total of 52 adults with abdominal aortoiliac aneurysm and prior 120-kVp single-energy CTA (SECTA) with 33 g iodine (standard dose) underwent follow-up dual-energy CTA (DECTA) with a 52% reduced iodine dose. Subjects were randomly assigned to a contrast medium protocol for DECTA examinations: one group (n = 26) received 16.2 g (270 mg I/mL) and the other (n = 26) received 16.0 g (320 mg I/mL). Two readers independently assessed SECTA and VMC DECTA datasets for image quality using a 5-point scale. Aortoiliac intravascular attenuation was measured, and ANOVA was used to compare measurements between VMC DECTA and SECTA images. In a subset of patients with DECTA after endovascular aortic repair, endoleak detection was evaluated on VMC images. Volume CT dose index, dose-length product, and size-specific dose estimate were compared between DECTA and SECTA. All DECTA examinations (n = 52) were rated diagnostic with image quality scores comparable to those of 120-kVp single-energy CTA (40 keV, 4.2-4.4; 50 keV, 4.6-4.8; SECTA, 4.4-4.5). Intravascular attenuation was uniform in all reduced-iodine DECTA examinations and was significantly higher on 40- and 50-keV images than on standard-iodine-dose SECTA images (720 ± 125 HU and 482 ± 82 HU vs 303 ± 65 HU) (p < 0.01). There was no difference in intravascular attenuation between the 16.2-g and the 16.0-g doses (p = 0.82). Sensitivity and specificity for endoleak detection were 78.9-94.7% and 100%. Total dose-length product was lower for DECTA (788 ± 166 mGy · cm) than for SECTA (1114 ± 468 mGy · cm). Low-energy VMC DECTA images (40 and 50 keV) acquired with two contrast protocols at approximately 50% reduced iodine dose (16.0 and 16.2 g) provide adequate intravascular attenuation and diagnostic quality for aortoiliac evaluation.


Obesity and the lung: 1. Epidemiology.
Obesity is the most common metabolic disease in the world and its prevalence has been increasing over several decades. The World Health Organization (WHO) predicts that, by 2015, around 700 million adults will be obese (at least 10% of the projected global population). This will be a huge health and economic burden with associated increases in diabetes, cardiovascular and musculoskeletal disease, and malignancy. While there has been little focus on the impact of obesity on respiratory disease, there are clear effects on pulmonary function and inflammation which will increase the prevalence and morbidity of lung disease. There is an inverse relationship between body mass index and forced expiratory volume in 1 s. Increases in body weight lead to worsening of pulmonary function. The reasons for this include the mechanical effects of truncal obesity and the metabolic effects of adipose tissue. Obesity is linked to a wide range of respiratory conditions including chronic obstructive pulmonary disease, asthma, obstructive sleep apnoea, pulmonary embolic disease and aspiration pneumonia. It is important for those providing care for people with respiratory disease to appreciate the impact of obesity and to provide appropriate advice for weight reduction. Healthcare planners should consider the impact of obesity for future resources in respiratory care.


Restriction endonuclease and nucleotide sequence analyses of molecularly cloned unintegrated avian tumor virus DNA: structure of large terminal repeats in circle junctions.
Avian tumor virus supercoiled DNA was isolated from infected quail tumor cells and molecularly cloned in pBR322. Four different recombinant clones denoted pATV-6, pATV-7, pATV-8, and pATV-9 were characterized in detail by restriction endonuclease mapping and by DNA sequencing. The results of these studies indicate that (i) the two large terminal repeats (LTRs) present in PATV-6, are different sizes, (ii) pATV-8 and pATV-9 contain only one LTR, (iii) pATV-7 contains an inversion of 0.6 kilobase in the env gene and a deletion of the U3 region and the src gene, and (iv) the src gene is deleted in pATV-6 and pATV-9. Circle formation from linear molecules was also examined in several of the clones by DNA sequencing through the circle joint. pATV-6 is an example of one class of circular molecules and contains a partially repeated LTR similar to that reported by Ju and Skalka (Cell 22:379-386, 1980). A second class of circles was exemplified by pATV-8 and pATV-9, which contain a single copy of the LTR with no base changes or deletions. This is in contrast to a class of circles containing a complete double LTR structure described by Swanstrom et al. (Proc. Natl. Acad. Sci. U.S.A. 78:124-128, 1981) and suggests that circles containing a single intact LTR may be formed by a homologous recombinational event in which an entire LTR or complementary regions from both LTRs are removed from the linear DNA molecule during circularization.


Fast detection and quantification of Plasmodium species infected erythrocytes in a non-endemic region by using the Sysmex XN-31 analyzer.
Due to increased travel from endemic countries, malaria occurs more frequently in non-endemic regions. It is a challenge for diagnostic laboratories in non-endemic countries to provide reliable results, as experience of staff is often limited to only a few cases per year. This study evaluated the diagnostic accuracy of the fully automated Sysmex XN-31 malaria analyzer in a routine diagnostic setting in a non-endemic region was evaluated. Samples from 112 patients suspected for malaria were examined by the Sysmex XN-31 analyzer to determine the absolute count of malaria-infected red blood cells count (MI-RBC/µL). Microscopic examination of both Quantitative Buffy Coat capillary tubes and thick and thin blood films were used as reference methods. Limits of blank (LoB), detection (LoD) and quantification (LoQ) were investigated using an in vitro Plasmodium falciparum culture. Nine hundred twenty samples of patients with RBC abnormalities were included to determine which RBC abnormalities trigger indeterminate or false positive results. No false positive nor false negative results were obtained for the examined patient samples suspected for malaria. For 3% of samples an indeterminate result by the XN-31 was obtained. The Passing-Bablok regression line for diagnostic accuracy of the parasitaemia was y = 39.75 + 0.7892 × showing a positive bias of about 21% when comparing the MI-RBC results to microscopy. The LoB, LoD and LoQ were calculated to be 4.7, 5.9, and 19.0 infected RBC/μL, respectively. From the 920 abnormal RBC samples collected, 4.6% resulted in a false positive MI-RBC result and almost half of the samples produced indeterminate results. These results were related to increases in nucleated red blood cells, reticulocytes and other abnormal RBC morphologies such as sickle cells. Based on the results, the XN-31 is a fast and reliable screening method in the detection and quantification of Plasmodium species in patients However, if an abnormal red blood cell morphology is present, the results of the XN-31 should be interpreted with caution as false positive results can be caused by interfering abnormal erythrocytes.


Biologic determinants of uveal melanoma metastatic phenotype: role of intermediate filaments as predictive markers.
The long-range goal of our research is to develop intervention strategies based on newly discovered biologic mechanisms responsible for the invasive dissemination of metastatic uveal melanoma. To accomplish this goal, we have focused on the biologic relevance of novel marker proteins contributing to the uveal melanoma metastatic phenotype. The expression of vimentin intermediate filaments (IFs), a mesenchymal marker, is typical of melanomas, whereas carcinomas typically express keratin IFs, which are markers for epithelia. Thus, cells that coexpress both IFs are regarded as "interconverted" in that they display both mesenchymal and epithelial phenotypes. Although the biologic functions of IFs have remained enigmatic, there is substantial support to suggest that the significance of vimentin/keratin coexpression is linked with poor patient outcome in cutaneous melanoma. Our data demonstrate that human uveal melanoma cell lines (isolated from primary choroidal or ciliary body melanomas and from foci of metastatic uveal melanoma to the liver), which contain predominant populations of cells that coexpress vimentin/keratins 8 and 18 (keratins 8,18) IFs, were 6-fold more invasive through collagenous extracellular matrices in vitro, compared with uveal melanoma cells expressing vimentin only, and were 8- to 13-fold more invasive than normal uveal melanocytes. Colocalization of vimentin/keratins 8,18 in cell cultures was corroborated by immunohistochemistry in histologic sections of tumors from which the cell lines were derived. Minor populations of these cells also coexpressed keratins 13 and 17. Experimental down-regulation of the predominant keratins 8,18 in the interconverted cells, using 16-mer antisense oligonucleotides, resulted in a significant decrease in the migratory ability of the cells-similar to levels achieved by cells positive only for vimentin. These findings provide justification for additional studies of the association between coexpression of IFs vimentin/keratins 8,18 and uveal melanoma metastasis.


Pollen counts in relation to the prevalence of allergic rhinoconjunctivitis, asthma and atopic eczema in the International Study of Asthma and Allergies in Childhood (ISAAC).
Although pollens are major allergens associated with allergic rhinoconjunctivitis and asthma, there is little information about the relative prevalence of these conditions in populations with different pollen exposures. The purpose of this study was to investigate the relationship between pollen exposure and allergic symptoms among children in different countries. An ecological analysis was conducted to see whether pollen exposure (pollen counts, and duration and severity of pollen seasons) is associated with symptoms of allergic rhinoconjunctivitis, asthma and atopic eczema in 28 centres within 11 countries (nine being in Europe). Data on the prevalence of symptoms in 13-14-year olds were based on the responses to the written questionnaires from the International Study of Asthma and Allergies in Childhood (ISAAC). The analysis was adjusted for gross national product and mean annual relative humidity. There was little relationship between pollen exposure and symptom prevalence, except for a significant inverse association between grass pollen counts and lifetime prevalence of the symptoms of allergic rhinitis (P=0.03). Almost all the regression coefficients were negative. The associations were even weaker and all non-significant when the analyses were conducted within countries, using a random intercept fixed slope model, but there was still no evidence of a positive association between pollen exposure and symptoms. There is a weak but consistent tendency for the prevalence of allergic symptoms to be inversely associated with pollen exposure. This finding accords with evidence from several countries, suggesting that the prevalence of hayfever and asthma tends to be lower in rural than in urban areas, and lowest among people living on farms. Exposure to allergenic pollen in early life does not appear to increase the risk of acquiring symptoms of respiratory allergy, and may even give some protection against them.


Epithelial adhesion mediated by pilin SpaC is required for Lactobacillus rhamnosus GG-induced cellular responses.
Lactobacillus rhamnosus GG is a widely used probiotic, and the strain's salutary effects on the intestine have been extensively documented. We previously reported that strain GG can modulate inflammatory signaling, as well as epithelial migration and proliferation, by activating NADPH oxidase 1-catalyzed generation of reactive oxygen species (ROS). However, how strain GG induces these responses is unknown. Here, we report that strain GG's probiotic benefits are dependent on the bacterial-epithelial interaction mediated by the SpaC pilin subunit. By comparing strain GG to an isogenic mutant that lacks SpaC (strain GGΩspaC), we establish that SpaC is necessary for strain GG to adhere to gut mucosa, that SpaC contributes to strain GG-induced epithelial generation of ROS, and that SpaC plays a role in strain GG's capacity to stimulate extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling in enterocytes. In addition, we show that SpaC is required for strain GG-mediated stimulation of cell proliferation and protection against radiologically inflicted intestinal injury. The identification of a critical surface protein required for strain GG to mediate its probiotic influence advances our understanding of the molecular basis for the symbiotic relationship between some commensal bacteria of the gut lumen and enterocytes. Further insights into this relationship are critical for the development of novel approaches to treat intestinal diseases.


Numerical optimization approach to modelling delamination and buckling of geometrically constrained structures.
Understanding what happens in terms of delamination during buckling of laminate materials is of importance across a range of engineering sectors. Normally concern is that the strength of the material is not significantly impaired. Carton-board is a material with a laminate structure and, in the initial creation of carton nets, the board is creased in order to weaken the structure. This means that when the carton is eventually folded into its three-dimensional form, correct folding occurs along the weakened crease lines. Understanding what happens during creasing and folding is made difficult by the nonlinear nature of the material properties. This paper considers a simplified approach which extends the idea of minimizing internal energy so that the effects of delamination can be handled. This allows a simulation which reproduces the form of buckling-delamination observed in practice and the form of the torque-rotation relation.


T3 surface molecules on adult T cell leukemia cells are modulated in vivo.
Cells from eight patients with adult T cell leukemia (ATL) and from four patients with non-ATL were examined to see if the T3 antigen of these cells could be modulated in vitro. We found a low density of T3 antigen and the presence of Tac antigen on cells from all patients with ATL. The density of T3 antigen on non-ATL cells was normal, and Tac antigen was not detected. Modulation of T3 antigen and an increase in Tac antigen-positive cells occurred when cells from patients with T4 non-ATL were cultured with OKT3 monoclonal antibody (mAb). Those changes in T3 antigen density and the appearance of Tac antigen-bearing cells by OKT3 mAb were not so marked when ATL cells were used. But the modulation of T3 antigen and the increase in Tac antigen-bearing cells by OKT3 mAb were closely related in cells from six ATL patients. These findings suggest that T3 T cell antigen receptor complexes on ATL cells are not functionally "frozen" by leukemic changes and might be modulated in vivo. In addition, modulation of T3 surface antigen on ATL cells was not induced by cultivation with human T cell leukemia virus type I particles and envelope proteins obtained by gene technology.


Paving the Way for Dose Banding of Chemotherapy: An Analytical Approach.
Background: In an interdepartmental cooperation, we investigated the feasibility and benefits of implementing dose banding of chemotherapy at our medical center. Based on this concept, chemotherapy doses are clustered into bands of similar dosage levels, thereby allowing the preproduction of frequently used standard doses of drugs, with sufficient physicochemical stability. Although established practice in the United Kingdom, there is little published evidence of its introduction elsewhere. Methods: We performed an analysis of local prescribing practice (22,310 chemotherapies) and identified gemcitabine, 5-fluorouracil, and carboplatin, among various others, as cytotoxic drugs suitable for dose banding. Results: First, we determined the physicochemical stability of the selected chemotherapy drugs during 12-weeks' storage by performing pH analysis and visual examination for color change or particles. No relevant changes were identified. Gemcitabine was selected for quantitative high-performance liquid chromatography analysis and we were able to show that ≥95% remained after 12 weeks' storage, in accordance with international guidelines. To simulate a worst case scenario, we performed microbiological stability testing of simulated cytotoxic compounding by replacing the cytotoxic drug with liquid media. Samples were incubated over defined storage time points (3, 6, and 12 weeks) and evaluated using the direct inoculation method. For the container integrity test, we deposited the samples into highly contaminated broth for 1 hour. Microbiological stability was demonstrated in both tests for the full storage period. Conclusions: Our data show that 12-weeks' storage of selected cytotoxic products is feasible from a microbiological perspective. Sterility of prepared products was maintained under extreme storage conditions. Gemcitabine content was in accordance with international guidelines after 12-weeks' storage. These results support the introduction of dose-banded gemcitabine products with the predicted advantages of optimized pharmacy workflow and reduced patient waiting times. We highlight the need for further research and consensus on the performance of purity analyses in dose-banded drug products.


Mutations outside the rifampicin resistance-determining region associated with rifampicin resistance in Mycobacterium tuberculosis.
Ninety-six percent of rifampicin resistance in Mycobacterium tuberculosis was shown to be associated with mutations inside the 81 bp rifampicin resistance-determining region (RRDR) located in the centre of the rpoB gene. The detection of rifampicin resistance by targeting the RRDR failed to match with a resistant phenotype in 4% of all cases. Our study aims to identify the mutations outside the RRDR that are associated with rifampicin resistance in M. tuberculosis. Among 50 rifampicin-resistant and 20 rifampicin-susceptible clinical isolates of M. tuberculosis, 2 of the rifampicin-resistant isolates did not harbour any known mutations in the RRDR. Sequencing analysis of the whole rpoB gene identified two rare mutations, V146F and I572F. A molecular structure model based on Thermus thermophilus RpoB revealed that both these substituted amino acids are located in close proximity to the rifampicin-binding pocket of the β-subunit. Substitutions of simple amino acids for bulky ones are likely to affect the protein-drug interaction. Cloning and transformation of the mutated rpoB gene into wild-type Mycobacterium smegmatis and M. tuberculosis successfully elevated the MIC of rifampicin and conferred the rifampicin resistance phenotype. Our study showed that amino acid positions 146 and 572 are associated with rifampicin resistance in M. tuberculosis in addition to the RRDR. Molecular assays for identifying rifampicin-resistant M. tuberculosis might be improved in terms of accuracy by including these two positions.


Optimization of animal manure vermicomposting based on biomass production of earthworms and higher plants.
The goal of this study was to optimize the mixture of swine manure (SM) and cattle manure (CM) used in the vermicomposting process, seeking to increase the manure biodegradation rate and enhance the biomass production of both earthworms and higher plants. To achieve this goal, physico-chemical parameters were determined to assess the final compost quality after 50 days of vermicomposting. The different manure ratios used to produce the composts (C) were as follows (SM:CM, % m/m basis): C1 100:0, C2 (75:25), C3 (50:50), C4 (25:75), and C5 (0:100). In addition, the earthworm biomass and the phytoproductivity of lettuce (Lactuca sativa L.) plants grown in mixtures (1:1) of natural soil and the most viable vermicomposts were investigated. The C1 and C2 compost compositions were associated with high earthworm mortality rates. The C3 compost provided the highest mineral concentrations and C5 showed the highest lettuce yield (wet biomass). The results verify that stabilized cattle manure is an excellent substrate for the vermicomposting process and that fresh swine manure must be mixed with pre-stabilized cattle manure to ensure an optimized vermicomposting process, which must be controlled in terms of temperature and ammonia levels. It is concluded that small livestock farmers could add value to swine manure by applying the vermicomposting process, without the need for high investments and with a minimal requirement for management of the biodegradation process. These are important technical aspects to be considered when circular economy principles are applied to small farms.