Pasteurization of human milk affects the miRNA cargo of EVs decreasing its immunomodulatory activity.
In this report, we evaluated the effect of the pasteurization (P) process of mother's own milk (MOM) on the miRNA content of extracellular vesicles (EVs) and its impact on innate immune responses. Differences in size or particle number were not observed upon pasteurization of MOM (PMOM). However, significant differences were observed in the EV membrane marker CD63 and miRNA profiles. miRNA sequencing identified 33 differentially enriched miRNAs between MOMEV and PMOMEV. These changes correlated with significant decreases in the ability of PMOMEV to modulate IL-8 secretion in intestinal Caco2 cells where only MOMEV were able to decrease IL-8 secretion in presence of TNFα. While EVs from MOMEV and PMOMEV were both able to induce a tolerogenic M2-like phenotype in THP-1 macrophages, a significant decrease in the transcript levels of IL-10 and RNA sensing genes was observed with PMOMEV. Together, our data indicates that pasteurization of MOM impacts the integrity and functionality of MOMEV, decreasing its EVs-mediated immunomodulatory activity. This data provides biomarkers that may be utilized during the optimization of milk processing to preserve its bioactivity.


Right-to-left shunts in lowlanders with COPD traveling to altitude: a randomized controlled trial with dexamethasone.
Right-to-left shunts (RLS) are prevalent in patients with chronic obstructive pulmonary disease (COPD) and might exaggerate oxygen desaturation, especially at altitude. The aim of this study was to describe the prevalence of RLS in patients with COPD traveling to altitude and the effect of preventive dexamethasone. Lowlanders with COPD [Global Initiative for Chronic Obstructive Lung Disease (GOLD) grades 1-2, oxygen saturation assessed by pulse oximetry (SpO2) >92%] were randomized to dexamethasone (4 mg bid) or placebo starting 24 h before ascent from 760 m and while staying at 3,100 m for 48 h. Saline-contrast echocardiography was performed at 760 m and after the first night at altitude. Of 87 patients (81 men, 6 women; mean ± SD age 57 ± 9 yr, forced expiratory volume in 1 s 89 ± 22% pred, SpO2 95 ± 2%), 39 were assigned to placebo and 48 to dexamethasone. In the placebo group, 19 patients (49%) had RLS, of which 13 were intracardiac. In the dexamethasone group 23 patients (48%) had RLS, of which 11 were intracardiac (P = 1.0 vs. dexamethasone). Eleven patients receiving placebo and 13 receiving dexamethasone developed new RLS at altitude (P = 0.011 for both changes, P = 0.411 between groups). RLS prevalence at 3,100 m was 30 (77%) in the placebo and 36 (75%) in the dexamethasone group (P = not significant). Development of RLS at altitude could be predicted at lowland by a higher resting pulmonary artery pressure, a lower arterial partial pressure of oxygen, and a greater oxygen desaturation during exercise but not by treatment allocation. Almost half of lowlanders with COPD revealed RLS near sea level, and this proportion significantly increased to about three-fourths when traveling to 3,100 m irrespective of dexamethasone prophylaxis.NEW & NOTEWORTHY The prevalence of intracardiac and intrapulmonary right-to-left shunts (RLS) at altitude in patients with chronic obstructive pulmonary disease (COPD) has not been studied so far. In a large cohort of patients with moderate COPD, our randomized trial showed that the prevalence of RLS increased from 48% at 760 m to 75% at 3,100 m in patients taking placebo. Preventive treatment with dexamethasone did not significantly reduce the altitude-induced recruitment of RLS. Development of RLS at 3,100 m could be predicted at 760 m by a higher resting pulmonary artery pressure and arterial partial pressure of oxygen and a more pronounced oxygen desaturation during exercise. Dexamethasone did not modify the RLS prevalence at 3,100 m.


Ten-year nodular sclerosis Hodgkin's disease and second malignancies.
Hodgkin's disease is one of the most curable cancers thanks to progress in radiotherapy and multi-drug chemotherapy regimens such as mechloretamine-vincristine-procarbazine-prednisone, best known as MOPP. However, long-term side-effects and treatment-induced second malignancies are of great concern. In our institution, 69 patients with nodular sclerosis Hodgkin's disease were treated over 10 years. Twenty-two per cent were stage I, 49% stage II, 23% stage III and 6% stage IV. Actuarial 10-year survival was 83% and actuarial relapse-free survival 61%. Six patients developed a second malignancy with a 10-year actuarial risk of 18%. All six cases occurred in the group treated with MOPP and extensive radiotherapy. Acute non-lymphoblastic leukemia occurred in three patients, preleukemia in two and non Hodgkin's lymphoma in one. In all of these patients, the results were quite poor. Overall survival was equally affected by Hodgkin's disease and by second malignancies. Since new multiple-drug chemotherapy regimens such as adriamycin-bleomycin-vinblastine-dacarbazine, known as ABVD, are equally effective and seem less likely to induce second hematologic malignancies, we suggest that MOPP should no longer be used as a first choice for the treatment of Hodgkin's disease, especially when in combination with radiation therapy.


Cellular immune activation in Gulf War veterans.
The etiology and pathology of illnesses related to the first Persian Gulf War are unclear. Among the constellation of symptoms noted in sick veterans, some, such as skin rashes, musculoskeletal pains, and neuropsychiatric problems, have been proposed to reflect an underlying immune dysfunction. In this study we explored the hypothesis that sickness following deployment to the Gulf in 1991 is associated with altered immune function, and we examine possible associated exposures. In particular, we focused on peripheral blood Th1/Th2 balance by measuring intracellular production of IFN-gamma, IL-2 (Th1), IL-4 (Th2), and IL-10 by CD4 T cells, using a nested case control study design within a large epidemiological survey. We compared symptomatic Gulf War veterans (sGWV) with well GWVs (wGWV), and a second control group of symptomatic veterans who served in Bosnia or were nondeployed military personnel of the same era. We found evidence for an altered immune status in sGWV in comparison to the other study groups. In particular, ongoing Th1-type immune activation was associated with multisymptom illness in GWVs, with sick veterans having significantly elevated levels of IFN-gamma and IL-2 producing CD4+ cells in the absence of in vitro stimulation compared with wGWVs (P = 0.01 and P =0.001). In vitro polyclonal activation revealed significantly elevated levels of IL-10 producing memory CD4 cells in sGWVs (P <0.001), but other cytokines were normal. In terms of possible exposures that might influence immune function, we found a trend for reduced levels of IFN-gamma producing cells after polyclonal activation with increasing numbers of vaccines administered (P <0.05) but no changes in other cytokines. These data show that multisymptom illness in Gulf War veterans is characterized by ongoing Th1-type immune activation and a biased generation of memory cells secreting the suppressor cytokine, IL-10.


The Influence of UV Radiation on the Degradation of Pharmaceutical Formulations Containing Quercetin.
The aim of this study was to assess the photostability of quercetin in the presence of anionic and nonionic polymeric gels with varied compositions of an added component-glycerol. The samples were irradiated continuously at constant temperature. The stability of quercetin in solution and incorporated into the gels was evaluated by an UV-Vis spectrophotometer. FTIR spectroscopy (Fourier-transform infrared spectroscopy) was used to detect the changes in the structure of quercetin depending on the polymer used in the gel, and on the exposure time. Photostabilization is an important aspect of quality assurance in photosensitive compounds. The decomposition rate of quercetin in the ionic preparation of polyacrylic acid (PAA) with glycerol was 1.952·10-3 min-1, whereas the absence of glycerol resulted in a decay rate of 5.032·10-4 min-1. The formulation containing non-ionic methylcellulose resulted in a decomposition rate of quercetin in the range of 1.679·10-3 min-1. The decay rate of quercetin under light influence depended on the composition of the gel. It was found that the cross-linked PAA stabilized quercetin and the addition of glycerol accelerated the photodegradation.


Diabetic neuropathy in elderly Type 2 diabetic patients: effects of insulin treatment.
To study the occurrence of diabetic neuropathy and the effect of insulin treatment in elderly Type 2 diabetic patients. In 38 patients and 20 controls symptoms and neurophysiological examinations including electroneurography, vibration perception and temperature discrimination thresholds were investigated. Patients were randomized to insulin (n = 18) or sulfonylurea (n = 16) treatment and were re-investigated after 1 year. Neuropathy was present in 21/38 patients (56%). It was asymptomatic in 17/38 (45%) and symptomatic in 4/38 (11%). The occurrence of neuropathy was less common in healthy controls, 3/20 (15%) (P < 0.01). Temperature discrimination thresholds was the test that most often revealed pathology. The metabolic control after 1 year was significantly improved in the insulin treated group and unchanged in the sulfonylurea treated group. There were no changes as regards occurrence of neuropathy between or within the two treatment groups after 1 year. Diabetic neuropathy is common among elderly Type 2 diabetic patients. It is mostly asymptomatic. Improvement was not seen after 1 year of insulin treatment.


The good and the bad faces of STAT1 in solid tumours.
Signal transducer and activator of transcription (STAT) 1 is part of the Janus kinase (JAK)/STAT signalling cascade and is best known for its essential role in mediating responses to all types of interferons (IFN). STAT1 regulates a variety of cellular processes, such as antimicrobial activities, cell proliferation and cell death. It exerts important immune modulatory activities both in the innate and the adaptive arm of the immune system. Based on studies in mice and data from human patients, STAT1 is generally considered a tumour suppressor but there is growing evidence that it can also act as a tumour promoter. This review aims at contrasting the two faces of STAT1 in tumourigenesis and providing an overview on the current knowledge of the underlying mechanisms or pathways.


A possible role of the cytochrome P450c17alpha gene (CYP17) polymorphism in the pathobiology of uterine leiomyomas from black South African women: a pilot study.
To investigate the role of the CYP17 gene promoter polymorphism in the pathobiology of uterine leiomyomas in African and Caucasian women. During a 6-month period, 145 Caucasian and black South African women undergoing hysterectomy were included prospectively. Blood samples were obtained for DNA analysis. Factors modifying the risk for uterine leiomyoma growth such as age, parity, age at last birth, weight, body mass index, menopausal status, cigarette smoking and oral contraceptive use were determined. The risk for leiomyoma development among black South African homozygous carriers of the CYP17 A2 allele was shown to be significantly increased when compared to women homozygous for the CYP17 A1 allele or to heterozygous women. Logistic regression analysis disclosed age, parity and CYP17 polymorphism to have an impact on the presence of uterine leiomyomas (p-values are, respectively, 0.0006, 0.0001 and 0.03) in black South African women. However, among Caucasian women, logistic regression analysis showed only intake of oral contraceptives to influence the formation of uterine leiomyomas (p = 0.03). This exploratory trial suggests that among African women, homozygous carriers of the CYP17 A2 allele expose their myometrium to a stronger estrogenic stimulation contributing to the pathobiology of uterine leiomyomas.


Argonaute-triggered visual and rebuilding-free foodborne pathogenic bacteria detection.
Foodborne pathogenic bacteria are recognized as the main causes of microbial contamination in food safety. Early screening and ultrasensitive detection of foodborne pathogenic bacteria is critical procedure to guarantee food safety. Argonaute is emerging as a new tool for detection owing to the programmability and high specificity. We reported a Novel and One-step cleavage method based on Argonaute by integrating Tag-specific primer extension and Exonuclease I (Exo I) for the first time, termed as NOTE-Ago. In this method, the invA of Salmonella typhi and nuc gene of Staphylococcus aureus were amplified using Tag-specific primer and the remaining primers were digested by Exo I. Then amplicons were served as the guide DNA for PfAgo. Consequently, the fluorophore-quencher reporter could be cleaved via PfAgo, resulting in changes in fluorescent intensity. With this strategy, target nucleic acid could be dexterously converted into fluorescent signals. The NOTE-Ago assay could detect 1 CFU/mL with a dynamic range from 1 to 108 CFU/mL. The satisfactory selectivity of NOTE-Ago assay further facilitated its application for detecting S. typhi- and S. aureus-contaminated food samples. This work enriches the toolbox of Argonaute-based detection and provides a one-step cleavage and rebuilding-free method for ultrasensitive detection of bacteria.


Response of urinary hydroxyproline to dietary protein and fasting in white-tailed deer.
The effects of dietary protein, fasting, and refeeding on urinary hydroxyproline of nine captive female white-tailed deer (Odocoileus virginianus) were examined from 23 February to 3 May 1984 in northern Minnesota. In the fasted group, mean hydroxyproline:creatinine (OHP:C) was greater (P less than 0.05) at week 4 compared to baseline at week 0. Between fasted deer and deer fed high protein-high energy (HPHE) and low protein-high energy (LPHE) diets, no difference in OHP:C ratios was detected during the initial 4 wk of the study. Urinary OHP:C ratios were significantly (P less than 0.05) greater in the fasted group during refeeding, concomitant with greater feed consumption and weight gain. There was also a significant (P less than 0.02) time effect in the fasted-refed group; OHP:C ratios increased during these two phases of the study. There was no difference between the HPHE and LPHE fed deer in renal OHP excretion. However, mean OHP:C ratios in yearlings (16.8 +/- 2.2) were greater (P less than 0.001) than in the adults (7.5 +/- 1.2) of those groups, indicating a higher collagen turnover rate. Urinary OHP:C shows potential as an indicator of growth and starvation, and the data presented may serve as reference values.


Growth of an intrapelvic pseudotumor associated with a metal-on-metal total hip arthroplasty after revision arthroplasty causing a femoral nerve neuropathy.
The development of pseudotumors is not uncommon with metal-on-metal total hip arthroplasty. Pseudotumors that dissect into the retroperitoneal space can cause symptoms of nerve compression. We describe a case of a 53-year-old male with a metal-on-metal total hip arthroplasty who developed mild symptoms of a femoral nerve neuropathy 6 years postoperatively. Revision arthroplasty to a ceramic-on-polyethylene articulation and debridement of the pseudotumor was performed. Postoperatively, the patient's femoral neuropathy progressed and a repeat magnetic resonance imaging showed an increase in size of the pseudotumor despite the removal of the offending metal-on-metal articulation. The patient subsequently underwent a laparoscopic excision of the retroperitoneal pseudotumor. By 17 months post laparoscopic excision of the pseudotumor, the patient's motor deficits resolved, however, sensory deficits persisted in the anteromedial thigh.


Primary solitary fibrous tumor (SFT) in the retroperitoneum.
Solitary fibrous tumor (SFT) is an infrequent but distinct neoplasm, which generally arises from submesothelial connective tissue in the pleura. SFT is rarely recognized in extrathoracic sites, and histologically identical conditions have also been reported in the retroperitoneum, although their pathophysiology has not been extensively investigated. We present four cases of primary SFT in the retroperitoneum, and review 37 similar cases in the previous literature. About 40% of patients were asymptomatic, and 19.2% and 15.4% presented with an abdominal mass and urinary symptoms, respectively. The tumor size ranged between 2 and 26 (mean 9.1) cm. Sixty-three percent of tumors showed nonspecific development with haphazard distribution of bland short spindle or polygonal cells with or without collagenous bundles and stromal hyalinization. In 22.0%, hemangiopericytomatous appearance was seen. About 15% of cases showed histologically malignant characteristics. The tumor cells were immunoreactive for vimentin in all cases, CD34 in 91% and Bcl-2 in 86%. All tumors were excised, and in 85.4% of cases, tumors did not recur postoperatively for 6 to 48 months. No significant difference was found between the recurrence rate of histologically benign and malignant cases. Cases positive for both CD34 and Bcl-2 had no recurrence. The identification of SFT in the retroperitoneum is of importance because histopathological indicators of malignancy are not necessarily associated with clinical malignant potential in many cases of retroperitoneal SFT. Retroperitoneal SFT showing typical pathological features with expression of CD34 and Bcl-2 is associated with a favorable outcome following excision.


Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells.
Accurate identification of the murine estrous cycle using vaginal exfoliative cytology is the initial and crucial step for controlled reproduction of this species. However, it is generally difficult to discriminate each stage of the cycle, and thus to select pro-estrous mice for mating. To increase the accuracy of identification of the pro-estrous stage, we re-evaluated the vaginal fold histology and modified the method of exfoliative cytology. Tissue fixation using methanol in Carnoy's solution but not paraformaldehyde, combined with Alcian blue staining but not the conventional Giemsa staining, resulted in better manifestation of mucosal cell layers in the vaginal epithelium just above the keratinized layer. This mucous layer in the fold histology was found to form specifically in the pro-estrous and late di-estrous stages, and the mucous cells exfoliated in smear samples only in the pro-estrous stage. This novel method was found, by a blinded test, to increase the rate of accurate identification of the pro-estrous stage compared to the conventional method (80% vs 50%). Consistent with this finding, the mating experiment with "pro-estrous" females selected by the novel method revealed a significantly higher success rate than that with the conventional method (78.0% vs 47.5%). Thus, our study demonstrates vaginal exfoliative mucous cells as a better potential marker to detect the "receptive" state of female mice that leads to an improved success rate of mating.


Eucalyptus oil nanoemulsions against eggs and larvae of Haemonchus contortus.
Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus.


Examining Associations of Sexual Attraction and Attitudes on Women's Disordered Eating Behavior.
We aimed to expand the scant research on disordered eating in women identifying same-sex sexual attractions. We used multiple linear regressions to explore potential mechanisms driving disordered eating--both explicit and implicit weight bias and heterosexism--in a cross-sectional, online-recruited community sample of women (N = 437). Participants endorsed a range of sexual attractions from exclusively opposite-sex (21.1%) to exclusively same-sex (19.5%) attraction. Findings revealed no associations between sexual attraction and disordered eating. Awareness of sociocultural norms valuing thinness accounted for disordered eating for all women, regardless of sexual attraction, and was influenced by attitudes regarding weight. Among women endorsing same-sex attractions, self-reported internalized heterosexism influenced disordered eating. Findings contradict long-held beliefs that same-sex attracted women are protected from disordered eating. They emphasize a universal risk, for all women, of sociocultural norms valuing thinness, as well as the risk of internalized heterosexism among same-sex attracted women.


The seasonal variation of physical activity and quality of life during the first wave of the COVID-19 pandemic in Finnish older golfers: A cross-sectional study.
The COVID-19 pandemic was associated with reduced physical activity and increased sedentary behavior. Golf is a health-enhancing outdoor exercise and is associated with low risk of viral transmission. The purpose of this study was to describe the seasonal differences in physical activity and quality of life during the first wave of COVID-19 pandemic in 2020 in Finnish older golfers. Older golfers (n = 325) from eight golf clubs retrospectively responded to a questionnaire in summer 2020 on their physical activity and golf activity in winter 2019/20 (pre-COVID-19 winter season) and summer season 2020. In addition, they reported their quality of life after the first wave of pandemic in summer 2020. To evaluate the seasonal differences in physical activity, life quality and its association with golf activity, the data were analyzed using the Mann-Whitney U test, Wilcoxon signed-rank test, Spearman's correlation test, and linear regression analysis. Despite COVID-19 restrictions, golfers' physical activity increased by 24% (p < 0.001) during the COVID-19 restrictions in the summer of 2020. Moderate physical activity increased by 37% (p < 0.001), walking activity increased by 26% (p < 0.001), and sitting decreased by 21% (p < 0.001) compared with the pre-COVID-19 winter season. The full 18-hole golf round activity was positively associated with moderate physical activity both in the summer and winter season, and with walking in the summer season. Over 90% of golfers reported good quality of life during the 2020 summer restrictions. In contrast to physical activity levels generally decreasing during the first wave of pandemic, physical activity levels in Finnish golfers increased, and these golfers reported a good quality of life. Golf can provide health enhancing physical activity and older golfers seems to be physically active throughout the year.


DNA methylation in adolescents with anxiety disorder: a longitudinal study.
Anxiety disorders (AD) typically manifest in children and adolescents and might persist into adulthood. However, there are still few data concerning epigenetic mechanisms associated with onset, persistence or remission of AD over time. We investigated a cohort of adolescents and young adults at baseline (age; 13.19 ± 2.38) and after 5 years and classified them according to the AD diagnosis and their longitudinal trajectories into 4 groups: (1) Typically Developing Comparisons (TDC; control group, n = 14); (2) Incident (AD in the second evaluation only, n = 11); (3) Persistent (AD in both evaluations, n = 14) and (4) Remittent (AD in the first evaluation only, n = 8). DNA methylation was evaluated with the Infinium HumanMethylation450 BeadChip from saliva samples collected at both evaluations. Gene set enrichment analysis was applied to consider biological pathways. We found decreased DNA methylation in TDC group while the chronic cases of AD presented hypermethylation in central nervous system development pathways. Moreover, we showed that this persistent group also presented hypermethylation while the other three groups were associated with hypomethylation in nervous system development pathway. Incidence and remission groups were associated with increased and decreased methylation in neuron development pathways, respectively. Larger studies are likely to detect specific genes relevant to AD.


Characterization of steroid hormone receptors with ion-exchange fast protein liquid chromatography.
Androgen, estrogen and progesterone receptors have been characterized with anion exchange Fast Protein Liquid Chromatography (FPLC) on a Mono Q column (Pharmacia). In the presence of sodium molybdate androgen receptors in cytosols from rat prostate, rat epididymis and calf uterus eluted as a single sharp peak at 0.32 M NaCl with recoveries of approx 90%. The molybdate-stabilized form of the androgen receptor from rat prostate was purified about 75-fold. The receptor containing FPLC-peak fractions sedimented in high salt (0.4 M KCl) linear sucrose gradients at 3.6 S (prostate) and at 4.6 S (epididymis and calf uterus) respectively. Multiple forms of the androgen receptor were present in cytosols from rat prostate prepared in the absence of sodium molybdate, probably due to proteolytic breakdown of the native form. Calf uterine estradiol and progesterone receptors prepared in the presence of sodium molybdate (20 mM) eluted from the Mono Q column at 0.32 M NaCl. The molybdate-stabilized forms of the oestradiol and progesterone receptors were purified approx 70-fold and 30-fold respectively. In the absence of molybdate the estradiol receptor dissociated into two major forms eluting at 0.23 M NaCl and 0.37 M NaCl. After heat induced transformation (30 min at 25 degrees C) of the estradiol receptor one major peak was eluted at 0.42 M NaCl, indicating a change in the surface charge of the estradiol receptor as a result of the 4 S to 5 S transformation. It is concluded that the FPLC anion exchange system is a powerful, fast tool for characterization and partial purification of steroid receptors. In addition this technique could be applied as a rapid procedure for the quantitative estimation of steroid receptors in small biological samples.


The liquid-liquid extractive fermentation of L-lactic acid in a novel semi-partition bioreactor (SPB).
Fermentation technology is commonly used as a mature process to produce numerous products with the help of micro-organisms. However, these organisms are sometimes inhibited by the accumulation of these products or their by-products. One route to circumvent this is via extractive fermentation, which combines the fermentation process with extraction. To facilitate this, novel bioreactor designs are required, such as the semi-partition bioreactor (SPB) which has been recently proposed for in-situ extractive fermentation. The latter combines a fermentation and an extraction unit into a single vessel using a mixer-settler principle. Where the bioproduct is produced in the mixer and removed continuous in the settler. As the SPB functionality is a subject of interest, this study builds on demonstrating different process conditions in the production of a sample bioprocess (lactic acid (LA)) which is susceptible to product inhibition. The results showed a 34.5 g/L LA concentration was obtained in the pH-controlled condition. While LA production can suffer from product inhibition, neutralizing agents can be easily used to curb inhibitory problems, however, the LA fermentation is a simple (and well-studied) example, which can demonstrate an alternative route to avoiding product inhibition (for systems which cannot be rescued using pH control). Hence, to replicate a scenario of product inhibition, two different process conditions were investigated, no pH control with no extraction (non-integrated), and no pH control with integrated extractive fermentation. Key findings showed higher LA concentration in integrated (25.10 g/L) as compared to the non-integrated (14.94 g/L) case with improved yield (0.75 gg-1 (integrated) versus 0.60 gg-1 (non-integrated)) and overall productivity (0.35 gL-1h-1(integrated) versus 0.20 gL-1h-1(non-integrated)) likewise. This is the first demonstration of an SP bioreactor, and shows how the reactor can be applied to improve productivity. Based on these results, the SPB design can be applied to produce any product liable to product inhibition.


Angiogenic factors and their assay: activity of formyl methionyl leucyl phenylalanine, adenosine diphosphate, heparin, copper, and bovine endothelium stimulating factor.
A number of chemically unrelated substances have been compared for their neovasculogenic activity by two different in vivo tests, and for their ability to induce cultured endothelial cell migration and proliferation. Formyl methionyl leucyl phenylalanine, copper ions, heparin, adenosine diphosphate, and low-molecular-weight bovine endothelium stimulating factor were all neovasculogenic by the corneal pocket assay. By the chorioallantoic membrane assay, copper ions and formyl methionyl leucyl phenylalanine were not detectably neovasculogenic. By the same assay it has been possible to demonstrate angiogenic activity in bovine endothelium stimulating factor that is distinguishable from copper ions. This bovine factor, in contrast to the other agents, induced marked endothelial cell migration and also proliferation and may belong to a special class of agents which represents direct acting angiogenic activity.


Combating Implant Infections: Shifting Focus from Bacteria to Host.
The widespread use of biomaterials to support or replace body parts is increasingly threatened by the risk of implant-associated infections. In the quest for finding novel anti-infective biomaterials, there generally has been a one-sided focus on biomaterials with direct antibacterial properties, which leads to excessive use of antibacterial agents, compromised host responses, and unpredictable effectiveness in vivo. This review sheds light on how host immunomodulation, rather than only targeting bacteria, can endow biomaterials with improved anti-infective properties. How antibacterial surface treatments are at risk to be undermined by biomaterial features that dysregulate the protection normally provided by critical immune cell subsets, namely, neutrophils and macrophages, is discussed. Accordingly, how the precise modification of biomaterial surface biophysical cues, or the incorporation of immunomodulatory drug delivery systems, can render biomaterials with the necessary immune-compatible and immune-protective properties to potentiate the host defense mechanisms is reviewed. Within this context, the protective role of host defense peptides, metallic particles, quorum sensing inhibitors, and therapeutic adjuvants is discussed. The highlighted immunomodulatory strategies may lay a foundation to develop anti-infective biomaterials, while mitigating the increasing threat of antibacterial drug resistance.


[Decreased bone mineral density in HIV-infected children receiving highly active antiretroviral therapy].
In developed countries, the introduction of highly active antiretroviral therapy (HAART) has greatly improved survival and quality of life in HIV-infected children. Nevertheless, worrisome metabolic and bone alterations are beginning to be observed. To evaluate the prevalence of alterations in bone mineral density and possible associated factors in a cohort of HIV-infected children receiving HAART. Bone dual energy X-ray absorptiometry (DEXA) was performed in 50 HIV-infected children in a tertiary hospital in Madrid after a median length of HAART of 54 months. Subsequently, the group with bone mineral loss was compared with the group without bone mineral loss. Forty percent of the children studied had decreased bone mineral density, of which 36 % had osteopenia (18/50) and 4 % had osteoporosis (2/50). No statistically significant differences were found between the two groups in any of the factors analyzed. The prevalence of decreased bone mineral density in our cohort of HIV-infected children receiving HAART is high. The etiology and factors associated with this alteration are still not well known.


t-Butyl pyridine and phenyl C-region analogues of 2-(3-fluoro-4-methylsulfonylaminophenyl)propanamides as potent TRPV1 antagonists.
A series of 2-substituted 6-t-butylpyridine and 4-t-butylphenyl C-region analogues of 2-(3-fluoro-4-methylsulfonamidophenyl)propanamides were investigated for hTRPV1 antagonism. The analysis of structure activity relationships indicated that the pyridine derivatives generally exhibited a little better antagonism than did the corresponding phenyl surrogates for most of the series. Among the compounds, compound 7 showed excellent antagonism toward capsaicin activation with Ki=0.1nM and compound 60S demonstrated a strong antiallodynic effect with 83% MPE at 10mg/kg in the neuropathic pain model. The docking study of 7S in our hTRPV1 homology model indicated that the interactions between the A/B-regions of 7S with Tyr511 and the interactions between the t-butyl and ethyl groups in the C-region of 7S with the two hydrophobic binding pockets of hTRPV1 contributed to the high potency.


Pallidal substrate of morphine-induced locomotion.
Bilateral microinjections of morphine hydrochloride (5.0; 7.5; 10.0 micrograms/0.5 microliters/side) or saline were infused into 3 different regions (dorsal, medial, ventral) of the rat globus pallidus, to examine their effects on locomotor activity. Locomotor activity of each rat was measured 45 min before and 90 min after saline or morphine pallidal microinjections. Morphine induced a dose-dependent increase in locomotion. This increase in locomotion was also significantly different between the 3 pallidal regions. Pretreatment with naloxone (1 mg/kg, sc) inhibited the morphine (7.5 micrograms) hyperlocomotion elicited from all three pallidal areas. The results suggest that the entire pallidum serves as substrate of morphine hyperlocomotion mediated by opiate receptors.


Cortisol and testosterone in hair as biological markers of systolic heart failure.
Congestive heart failure (CHF) is associated with increased stress and alterations in metabolism, favoring catabolism over anabolism. Hormonal profiles of patients with heart failure have been assessed using serum and saliva as matrices, which are only point measurements and do not provide long-term information. Scalp hair is a novel matrix that allows for measurement of hormones over a period of several months. We aimed to evaluate whether levels of cortisol and testosterone and their ratio (C/T) in hair correlate with severity of heart failure. We conducted a prospective study in ambulatory male patients with a left ventricular ejection fraction (LVEF)≤40%. Hormone levels were measured using immunoassays in the proximal 2 cm of hair (representing approximately two months of systemic hormone exposure). Primary endpoints included the correlation of hair cortisol, testosterone, and C/T levels with the New York Heart Association (NYHA) class, LVEF, exercise capacity and NT-proBNP. The 44 CHF patients had a median hair level (range) of cortisol of 207 (117.7-1277.3)ng/g. Hair cortisol levels correlated positively with NYHA class (r=0.48, p=0.001) and negatively with treadmill stress test performance, (r=-0.37, p<0.05). The hair testosterone was 5.17 (2.39-24.64)ng/g and the C/T ratio was 39.89 (12.98-173.73). No associations were found between hair testosterone and C/T ratio and heart failure severity; however, the C/T ratio was higher in patients who required a CHF-related hospitalization than in patients who did not require this in the year following the inclusion in the study. Hair cortisol levels correlate with heart failure severity as assessed by the NYHA class and exercise capacity, while hair testosterone and C/T levels do not correlate with heart failure severity.


Using robotic mechanical perturbations for enhanced balance assessment.
Balance impairment is critical for many patient groups such as those with neural and musculoskeletal disorders and also the elderly. Accurate and objective assessment of balance performance has led to the development of several indices based on the measurement of the center of pressure. In this study, a robotic device was designed and fabricated to provide controlled and repeatable mechanical perturbations to the standing platform of the user. The device uses servo-controlled actuators and two parallel mechanisms to provide independent rotations in mediolateral and anterior-posterior directions. The device also provides visual feedback of the center of pressure position to the user. Functional tests were run and showed that the device is able to provide an appropriate dynamics (time constant of 0.19 s and bandwidth of 0.85 Hz) for the two motions. The efficacy of the device on the balance assessment was then evaluated experimentally. Ten healthy subjects performed a balance task with and without perturbations and seven center of pressure indices were measured. It was shown that the sensitivity of the indices to the user's performance was statistically increased in all indices particularly in anterior/posterior direction when the mechanical perturbations were present.


Brucellosis in camels (Camelus dromedarius) in Darfur, Western Sudan.
In a field outbreak of brucellosis in 21 camels mixed with cattle, sheep and goats, five camels, three of which showed clinical signs, were serologically positive. In a subsequent abattoir survey of apparently healthy camels, six animals were seropositive, albeit with titres that tended to be lower than those found in the field outbreak. Of the six seropositive slaughtered camels, five were shown to have lymph nodes (prescapular and supramammary) infected with brucellae (Brucella melitensis biovar 3, two camels; Brucella abortus biovar 6, three camels). Infection of camels with B. abortus biovar 6 had not previously been reported. Infection of the supramammary lymph nodes presents a potential hazard to those who consume raw camels' milk, a common practice in nomadic camel owners.


The trigeminal sensory innervation to the middle ear, eustachian tube, and pharynx: a study by the horseradish peroxidase tracer method.
The trigeminal sensory innervations to the middle ear, the eustachian tube, and the pharynx were examined using the horseradish peroxidase (HRP) method in cats. After HRP application to the middle ear and the eustachian tube, HRP-labeled cells were found mainly in the posterolateral portion and the anteromedial portion of the trigeminal ganglion, respectively. In the case of injection into the epipharynx and the mesopharynx, HRP-labeled cells were found in the anteromedial portion and the posterolateral portion, respectively. HRP-labeled cells were hardly observed when HRP was applied to the hypopharynx. These results indicate that trigeminal nerve sensory fibers branch to the middle ear, including the eustachian tube, epipharynx, and mesopharynx.


(E)-6-(4-Chloro-phen-yl)-4-[(2-cyano-3-phenyl-all-yl)sulfan-yl]-2,2-difluoro-3-phenyl-1,3,2-oxaza-borinin-3-ium-2-uide.
In the title compound, C25H18BClF2N2OS, the characteristic B-N and B-O bond lengths are 1.571 (3) and 1.458 (3) Å, respectively. The phenyl rings form dihedral angles of 83.1 (1) and 64.6 (1)° with the chloro-phenyl ring. In the crystal, weak C-H⋯N, C-H⋯F, C-H⋯π and π-π inter-actions [centroid-centroid distances 3.877 (6) Å between the chloro-phenyl rings of neighbouring mol-ecules] held mol-ecules together, forming ladders along the b axis.


Bayesian estimation of the parameter of Maxwell-Mukherjee Islam distribution using assumptions of the Extended Jeffrey's, Inverse-Rayleigh and Inverse-Nakagami priors under the three loss functions.
A three-parameter Maxwell-Mukherjee Islam distribution was proposed by applying Maxwell generalized family of distributions introduced by Ishaq and Abiodun [17]. The probability density and cumulative distribution functions of the proposed distribution were defined. The validity test was derived from its cumulative distribution function. The study aimed to obtain a Bayesian estimation of the scale parameter of Maxwell-Mukherjee Islam distribution by using assumptions of the Extended Jeffrey's (Uniform, Jeffrey's and Hartigan's), Inverse-Rayleigh and Inverse-Nakagami priors under the loss functions, namely, Squared Error Loss Function (SELF), Precautionary Loss Function (PLF) and Quadratic Loss Function (QLF), and their performances were compared. The posterior distribution under each prior and its corresponding loss functions was derived. The performance of the Bayesian estimation was illustrated from the basis of quantile function by using a simulation study and application to real life data set. For different sample sizes and parameter values, the QLF and SELF under Jeffrey's and Hartigan's priors produced the same estimates, bias and Mean Squared Error (MSE) just as we observed in their mathematical derivatives. Similarly, the SELF, PLF and QLF under Inverse-Rayleigh and Inverse-Nakagami priors provided the same performance when some parameter values are equal. For some parameter values, the QLF under Inverse-Nakagami and Inverse-Rayleigh priors produced the least values of MSE. In the application to real life data set, the QLF and SELF under Jeffrey's and Hartigan's priors; the SELF, PLF and QLF under Inverse-Rayleigh and Inverse-Nakagami priors provided similar results as observed in the simulation study. Therefore, the study concluded that the QLF under Inverse-Rayleigh and Inverse-Nakagami priors could effectively be used in the estimation of scale parameter of Maxwell-Mukherjee Islam distribution using Bayesian approach.


Derivation of haploid embryonic stem cells from mouse embryos.
Most animals are diploid, but haploid-only and male-haploid (such as honeybee and ant) species have been described. The diploid genomes of complex organisms limit genetic approaches in biomedical model species such as mice. To overcome this problem, experimental induction of haploidy has been used in fish. Haploid development in zebrafish has been applied for genetic screening. Recently, haploid pluripotent cell lines from medaka fish (Oryzias latipes) have also been established. In contrast, haploidy seems less compatible with development in mammals. Although haploid cells have been observed in egg cylinder stage parthenogenetic mouse embryos, most cells in surviving embryos become diploid. Here we describe haploid mouse embryonic stem cells and show their application in forward genetic screening.


Sociosexual stimuli and gonadotropin-releasing hormone/luteinizing hormone secretion in sheep and goats.
Sociosexual stimuli have a profound effect on the physiology of all species. Sheep and goats provide an ideal model to study the impact of sociosexual stimuli on the hypothalamic-pituitary-gonadal axis because we can use the robust changes in the pulsatile secretion of luteinizing hormone as a bioassay of gonadotropin-releasing hormone secretion. We can also correlate these changes with neural activity using the immediate early gene c-fos and in real time using changes in electrical activity in the mediobasal hypothalamus of female goats. In this review, we will update our current understanding of the proven and potential mechanisms and mode of action of the male effect in sheep and goats and then briefly compare our understanding of sociosexual stimuli in ungulate species with the "traditional" definition of a pheromone.


Bacterial contamination of the blood compartment originating from the dialysate in haemodialysers.
Micro-organisms originating from the dialysate compartment invaded the blood compartment of ' Rhodial RP6 ' haemodialysers in 16 out of 639 dialyses investigated (2.5 per cent). Analysis of the data suggested that the probable access of bacteria to the blood compartment was by way of minor defects in the dialysis membrane. The patients experienced no obvious symptoms or signs of sepsis which could be ascribed to the presence of micro-organisms in the blood compartment.


Enhancing Colorectal Cancer Radiation Therapy Efficacy using Silver Nanoprisms Decorated with Graphene as Radiosensitizers.
Metal nanoparticles have significant interaction cross-sections with electromagnetic waves due to their large surface area-to-volume ratio, which can be exploited in cancer radiotherapy to locally enhance the radiation dose deposition in tumors. We developed a new type of silver nanoparticle composite, PEGylated graphene quantum dot (GQD)-decorated Silver Nanoprisms (pGAgNPs), that show excellent in vitro intracellular uptake and radiosensitization in radiation-sensitive HCT116 and relatively radiation-resistant HT29 colorectal cancer cells. Furthermore, following biodistribution analysis of intravenously injected nanoparticles in nude mice bearing HCT116 tumors radiosensitization was evaluated. Treatment with nanoparticles and a single radiation dose of 10 Gy significantly reduces the growth of colorectal tumors and increases the survival time as compared to treatment with radiation only. Our findings suggest that these novel nanoparticles offer a promising paradigm for enhancing colorectal cancer radiation therapy efficacy.


Volumetric modulated arc therapy for nasopharyngeal carcinoma: a dosimetric comparison with TomoTherapy and step-and-shoot IMRT.
Volumetric modulated arc therapy (VMAT), a novel technique, employs a linear accelerator to conduct dynamic modulation rotation radiotherapy. The goal of this study was to compare VMAT with helical tomotherapy (HT) and step-and-shoot intensity-modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC) patients with regard to the sparing effect on organs at risk (OARs), dosimetric quality, and efficiency of delivery. Twenty patients with NPC treated by HT were re-planned by VMAT (two arcs) and IMRT (7-9 fields) for dosimetric comparison. The target area received three dose levels (70, 60, and 54 Gy) in 33 fractions using simultaneous integrated boosts technique. The Philips Pinnacle Planning System 9.0 was adopted to design VMAT, using SmartArc as the planning algorithm. For a fair comparison, the planning target volume (PTV) coverage of the 3 plans was normalized to the same level. Dosimetric comparisons between VMAT, HT, and IMRT plans were analyzed to evaluate (1) coverage, homogeneity, and conformity of PTV, (2) sparing of OARs, (3) delivery time, and (4) monitor units (MUs). The VMAT, HT, and IMRT plans had similar PTV coverage with an average of 96%. There was no significant difference between VMAT and HT in homogeneity, while the homogeneity indices of VMAT (1.06) and HT (1.06) were better than IMRT plans (1.07, p<0.05). HT plans provided a better conformity index (1.17) than VMAT (1.28, p=0.01) and IMRT (1.36, p=0.02). When compared with IMRT, VMAT and HT had a better sparing effect on brain stem and spinal cord (p<0.05). The effect of parotid sparing was similar between VMAT (mean=26.3 Gy) and HT (mean=27.5 Gy), but better than IMRT (mean=31.3 Gy, p<0.01). The delivery time per fraction for VMAT (5.7 min) were much lower than for HT (9.5 min, p<0.01) and IMRT (9.2 min, p<0.01). Our results indicate that VMAT provides better sparing of normal tissue, homogeneity, and conformity than IMRT, and shorter delivery time than HT.


The muscarinic M1 agonist xanomeline increases soluble amyloid precursor protein release from Chinese hamster ovary-m1 cells.
The functionally selective M1 agonist xanomeline, which is currently undergoing clinical trials as a therapy for Alzheimer's disease, was compared to the muscarinic agonist carbachol for effects on secretion of soluble amyloid precursor protein (APPs) from Chinese hamster ovary cells transfected with the human m1 receptor (CHO-m1). Release of APPs from CHO-m1 cells was increased maximally (4-10 fold) by 100 microM carbachol (EC50 = 11 microM) and by 100 nM xanomeline (EC50 = 10 nM). Stimulation of APPs secretion by xanomeline and carbachol was blocked by preincubation with 1 microM atropine. Carbachol did not stimulate APPs secretion from non-transfected CHO cells. Pilocarpine at 1 mM also increased APPs release. The efficacy of carbachol, xanomeline and pilocarpine for stimulating APPs secretion did not differ significantly. Activation of protein kinase C (PKC) in m1 transfected cell lines by 1 microM phorbol dibutyrate (PDBu) increased APPs release, and this was inhibited 97% by the PKC inhibitor bisindolemalemide. The PKC inhibitor decreased xanomeline and carbachol-stimulated APPs secretion by only 25-30%. These results demonstrate that xanomeline increased APPs release by activation of m1 muscarinic receptors and support the possibility that cholinergic replacement therapy for Alzheimer's Disease may reduce amyloid deposition.


Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. BET 3: can pregabalin effectively diminish acute herpetic pain and reduce the incidence of post-herpetic neuralgia?
A short cut review was carried out to establish whether pregabalin can reduce acute herpetic pain and reduce post herpetic neuralgia. 48 papers were found using the reported searches, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper are tabulated. It is concluded that pregabalin does not seem to decrease the intensity of pain related to acute herpes zoster. Moreover, it does not decrease the incidence of post herpetic neuralgia. More research is needed on this topic to clarify this issue [corrected].


Aortic valve exposure through a combined right atrial-ascending aortic approach in redo cases.
The expanded use of tissue valves in the aortic position lead to an increased number of reoperations in cases o valve failure. The approach to an aortic prosthesis can be difficult because of heavy adhesions, especially if biological glue was used in the first procedure and the interval between the first and second operation is short.


Linkage of interactions in sickle hemoglobin fiber assembly: inhibitory effect emanating from mutations in the AB region of the alpha-chain is annulled by a mutation at its EF corner.
The AB and GH regions of the alpha-chain are located in spatial proximity and contain a cluster of intermolecular contact residues of the sickle hemoglobin (HbS) fiber. We have examined the role of dynamics of AB/GH region on HbS polymerization through simultaneous replacement of non-contact Ala(19) and Ala(21) of the AB corner with more flexible Gly or rigid alpha-aminoisobutyric acid (Aib) residues. The polymerization behavior of HbS with Aib substitutions was similar to the native HbS. In contrast, Gly substitutions inhibited HbS polymerization. Molecular dynamics simulation studies of alpha-chains indicated that coordinated motion of AB and GH region residues present in native (Ala) as well as in Aib mutant was disrupted in the Gly mutant. The inhibitory effect due to Gly substitutions was further explored in triple mutants that included mutation of an inter-doublestrand contact (alphaAsn(78) --> His or Gln) at the EF corner. Although the inhibitory effect of Gly substitutions in the triple mutant was unaffected in the presence of alphaGln(78), His at this site almost abrogated its inhibitory potential. The polymerization studies of point mutants (alphaGln(78) --> His) indicated that the inhibitory effect due to Gly substitutions in the triple mutant was synergistically compensated for by the polymerization-enhancing activity of His(78). Similar synergistic coupling, between alphaHis(78) and an intra-double-strand contact point (alpha16) mutation located in the AB region, was also observed. Thus, two conclusions are made: (i) Gly mutations at the AB corner inhibit HbS polymerization by perturbing the dynamics of the AB/GH region, and (ii) perturbations of AB region (through changes in dynamics of the AB/GH region or abolition of a specific fiber contact site) that influence HbS polymerization do so in concert with alpha78 site at the EF corner. The overall results provide insights about the interaction-linkage between distant regions of the HbS tetramer in fiber assembly.


Influence of sub valvular pathology on immediate results and follow up events of Inoue balloon mitral valvotomy.
We evaluated the influence of sub valvular pathology (SVP) on the immediate results and follow up events of Inoue Balloon Mitral Valvotomy (IBMY) in 206 patients with severe SVP (Group I) and compared their outcome with 206 age and sex matched patients selected from the rest of 619 patients having mild/moderate SVP (Group II). Pre-procedure echocardiographic recordings were reviewed and mitral valve morphology was evaluated using U.S. California Score. The severe SVP group had lower mitral valve areas (MVA) (0.7 cm2 vs. 0.8 cm2) and higher mean pulmonary artery pressure (MPAP) (46.3+/-16.9 mmHg vs. 40.7+/-16.25 mmHg) and mean pulmonary capillary wedge pressure (PCWP) (27.5+/-7.3 mmHg vs. 25.7+/-8.0 mmHg) (p<0.001). IBMV was done using standard technique. The procedure was technically successful in 192/206 patients (93.2%) in group I and 187/206 (91%) in group II (p=ns). The mean transmitral gradient decreased from 24.8+/-7.6 mmHg to 7.46+/-3.4 mmHg while mean PCWP fell from 27.5+/-7.3 mmHg to 12.2+/-5.6 mmHg and MPAP fell from 46.3+/-16.9 mmHg to 23.6+/-12.2 mmHg (p=<0.001). MVA increased from 0.7+/-0.2 cm2 to 1.7+/-0.4 cm2 (p=<0.001). Severe mitral regurgitation (MR) occurred in 2 patients out of which one patient, who had associated coronary artery disease, died post operatively, and moderate MR occurred in 8 patients. The results achieved in patients with severe SVP were not statistically different from those with mild/moderate SVP. The benefits achieved immediate post IBMV were sustained in 184 patients with severe SVP who were available for follow up at a mean duration of 15.2 months (range 3 months to 51 months). Thus IBMV is safe and effective in patients with severe SVP. This group of patients with severe SVP are more hemodynamically deranged pre-BMV and also achieve better hemodynamic benefit compared to those with mild/moderate SVP. Severe SVP does not have any adverse effect either on immediate results (success/occurrence of MR) or on intermediate term follow up.


Serum profiles of pro-inflammatory and anti-inflammatory cytokines in non-hospitalized patients with mild/moderate COVID-19 infection.
This study attempted to explore pro-inflammatory and anti-inflammatory responses in patients with mild/moderate coronavirus disease 19 (COVID-19). Eight pro-inflammatory (IL-1α, IL-1β, IL-12, IL-17A, IL-17E, IL-31, IFN-γ and TNF-α) and three anti-inflammatory (IL-1Ra, IL-10 and IL-13) cytokines, as well as two chemokines (CXCL9 and CXCL10), were analyzed in the serum from ninety COVID-19 patients and healthy controls. Cytokine/chemokine levels were measured using enzyme-linked immunosorbent assay kits. Results revealed that IL-1α, IL-1β, IL-10, IL-12, IL-13, IL-17A, IL-31, IFN-γ, TNF-α and CXCL10 were significantly higher in patients than in controls, while IL-1Ra levels were significantly lower in patients. IL-17E and CXCL9 levels showed no significant differences between patients and controls. Seven cytokines/chemokines recorded an area under the curve greater than 0.8: IL-12 (0.945), IL-17A (0.926), CXCL10 (0.909), IFN-γ (0.904), IL-1α (0.869), TNF-α (0.825) and IL-10 (0.821). As indicated by the odds ratio, elevated levels of nine cytokines/chemokines were associated with an increased risk of COVID-19: IL-1α (19.04), IL-10 (5.01), IL-12 (43.66), IL-13 (4.25), IL-17A (16.62), IL-31 (7.38), IFN-γ (13.55), TNF-α (12.00) and CXCL10 (11.18). Only one positive (IL-17E with TNF-α) and six negative (IL-1β, IL-17A and IL-17E with CXCL9, IL-10 with IL-17A, and IL-1β and IL-17A with CXCL10) correlations were found between these cytokines/chemokines. In conclusion, pro-inflammatory (IL-1α, IL-1β, IL-12, IL-13, IL-17A, IL-31, IFN-γ, TNF-α and CXCL10) and anti-inflammatory (IL-10 and IL-13) cytokines/chemokines were up-regulated in the serum of patients with mild/moderate COVID-19. Their potential as biomarkers for diagnosis and prognosis is suggested and the association with COVID-19 risk is indicated to give more insight on COVID-19 immunological responses among non-hospitalized patients.


Safety and immunogenicity of the malaria candidate vaccines FP9 CS and MVA CS in adult Gambian men.
We assessed the safety and immunogenicity of prime-boost vectors encoding the Plasmodium falciparum circumsporozoite (CS) protein expressed either in the attenuated fowl-pox virus (FP9) or modified vaccinia virus Ankara (MVA). Thirty-two adult Gambians in groups of four to eight received one, two or three doses of FP9 CS and/or MVA CS. No serious adverse event was observed following vaccination. The most immunogenic regimen was two doses of FP9 followed by a single dose of MVA 4 weeks later (an average of 1000 IFN-gamma spot forming units/million PBMCs). This level of effector T-cell responses appears higher than that seen in previously reported studies of CS-based candidate malaria vaccines.


Robot-assisted Mitrofanoff and Malone antegrade continence enema reconstruction using divided appendix.
Fecal soiling or intractable constipation frequently occurs in association with urinary incontinence in children with varying pathologic conditions. To manage double incontinence or the combination of wetting and severe constipation, a Mitrofanoff conduit and a channel for antegrade continence enema is constructed using the appendix by open surgery. We present a video of the first case of robot-assisted laparoscopy for simultaneous reconstruction of a continent urinary diversion using the Mitrofanoff principle and a Malone antegrade continent enema using the divided appendix.


Detection of 3-nitropropionic acid and cytotoxicity inMucor circinelloides.
Mucorales are regarded as the aetiological agents of Mucormycosis. Their capabilities to produce mycotoxins are not profoundly investigated, in contrast to those of the fungi from the generaPenicillium, Aspergillus, orFusarium. The aim of this study was to isolate and identify fungi of the order Mucorales and investigate mycotoxins production. Twelve samples of visibly moulded grass silage and eight samples of damaged whole crop maize silage were analysed. Malt extract agar plates were used for sub cultivation. Three fungal species of the order Mucorales were isolated from grass silage, which were identified by their macro-and micro-morphology asAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer. The cytotoxicity ofMucor circinelloides extract was analysed using the cytotoxicity test (MTT assay) and the result, showed a low cytotoxicity. Additionally extracts fromAbsidia corymbifera, Mucor circinelloides andRhizopus stolonifer were tested for mycotoxin-production using an LC/MS/MS-based multimycotoxin method. 3-nitropropionic acid was detected in the culture extract ofMucor circinelloides.


Mitochondria from the Outside in: The Relationship Between Inter-Organelle Crosstalk and Mitochondrial Internal Organization.
A fundamental role of membrane-bound organelles is the compartmentalization and organization of cellular processes. Mitochondria perform an immense number of metabolic chemical reactions and to efficiently regulate these, the organelle organizes its inner membrane into distinct morphological domains, including its characteristic cristae membranes. In recent years, a structural feature of increasing apparent importance is the inter-connection between the mitochondrial exterior and other organelles at membrane contact sites (MCSs). Mitochondria form MCSs with almost every other organelle in the cell, including the endoplasmic reticulum, lipid droplets, and lysosomes, to coordinate global cellular metabolism with mitochondrial metabolism. However, these MCSs not only facilitate the transport of metabolites between organelles, but also directly impinge on the physical shape and functional organization inside mitochondria. In this review, we highlight recent advances in our understanding of how physical connections between other organelles and mitochondria both directly and indirectly influence the internal architecture of mitochondria.


Cholesterol depletion enhances adrenergic signaling in cardiac myocytes.
Cardiac myocytes endogenously express α and β adrenergic receptors, prototypes of the G-protein coupled receptor superfamily. Depending upon the dose of norepinephrine (agonist) exposure, hypertrophy and apoptosis are initiated by differential induction of two discrete constituents of the transcription factor AP-1, i.e., FosB and Fra-1. We explored differential adrenergic signaling as a paradigm for understanding how cholesterol dictates cells to choose hypertrophy or apoptosis. For this, we used fosB and fra-1 promoter-reporter constructs for monitoring adrenergic signaling. We show that cholesterol depletion enhances norepinephrine-mediated signaling in cardiac myocytes. Importantly, this increased signaling is reduced to original level upon cholesterol replenishment. We used specific ligands for α and β adrenergic receptors and show that the enhanced signaling upon cholesterol depletion is a combined effect of both α and β adrenergic receptors. These results constitute the first report demonstrating the effect of cholesterol on adrenergic signaling using a direct end-point gene expression.


Advances in the development of nucleoside and nucleotide analogues for cancer and viral diseases.
Nucleoside analogues have been in clinical use for almost 50 years and have become cornerstones of treatment for patients with cancer or viral infections. The approval of several additional drugs over the past decade demonstrates that this family still possesses strong potential. Here, we review new nucleoside analogues and associated compounds that are currently in preclinical or clinical development for the treatment of cancer and viral infections, and that aim to provide increased response rates and reduced side effects. We also highlight the different approaches used in the development of these drugs and the potential of personalized therapy.


Regulation of the prorenin - angiotensin system by oxygen and miRNAs; parallels between placentation and tumour development?
Tissue renin-angiotensin systems (RASs) are involved in tissue growth and development as they are important regulators of angiogenesis, cell proliferation and migration. The placental RAS is most highly expressed in early gestation, at a time when the oxygen tension within the conceptus is reduced, and plays a key role in placental growth and development. Similar to the placenta, tumour development relies on proliferation, angiogenesis and invasion in order to grow and metastasize. The RAS is known to be upregulated in a variety of solid tumours, including ovarian, endometrial, cervical, breast and prostate. This review explores the roles of oxygen and microRNAs in regulating the normal expression of the placental RAS, providing insight into regulation of its development as well as the development of disease states in which the RAS is overexpressed. We propose that the placental RAS is downregulated by microRNAs that are suppressed during the physiologically normal 'hypoxic' phase of early placentation. Suppression of these miRNAs allows the placental RAS to stimulate placental growth and angiogenesis. We propose that similar mechanisms may be at play in solid tumours, which are characterised by hypoxia.


Assessment of land use effect, mapping of human health risks and chemometric analysis of potential toxic elements in topsoils of Aran-o-Bidgol, Iran.
This study examines topsoil contamination in Aran-o-Bidgol urban region of central Iran, with a focus on potentially toxic elements (PTEs). A total of 135 topsoil samples in different land types were characterized, ranging from areas with agricultural farms, desert, industrial and residential activity, and brick kilns. The average concentrations of Cd, Pb, Cu, Ni, Cr, Co, Fe, Zn, and Mn were 0.72, 11.41, 14.82, 29.87, 51.13, 106.69, 8741.87, 48.59, and 346.42 mg kg-1, respectively, which all exceed the local background levels. The results reveal that land use significantly affected PTE concentrations. Cr, Co, Mn, and Fe concentrations in soils of residential and brick kiln areas were especially high. In contrast, concentrations of Cu, Ni, and Zn were higher in agricultural and residential areas. Risk assessment analysis showed that the sum of toxic units for PTEs for brick kilns (1.72), residential (1.82), and agricultural (1.79) areas exceeded those of other land types and that Ni and Cr contributed the most to the high toxic risk index values. Both carcinogenic and non-carcinogenic risk indices of PTEs in soils were within an acceptable limit, except for the cancer risk of Ni (3.52E-04) and Cr (3.00E-04) among children. The spatial hazard index and carcinogenic health risk of PTEs showed that samples from the southwestern parts of the study area might pose significant health problems to adults and children. This study demonstrates how combining different techniques can help spatially characterize PTE accumulation and protect populations at risk.


Dataset of antibody variable region sequence features inferred from a respiratory syncytial virus fusion protein-specific B cell receptor repertoire induced by natural infection of a healthy adult.
Respiratory syncytial virus (RSV) is the primary cause for acute lower respiratory syndrome in children younger than 5 years. Research on B cell repertoires and antibodies binding the RSV fusion protein (RSV F) is of major interest in the development of potential vaccine candidates and therapies. B cell receptors (BCRs) which have higher affinities for a specific antigen are preferentially selected for B cell clonal expansion in germinal center reactions. Consequently, antigen-specific BCR repertoires share common features, as for instance preferential variable gene usage, variable region mutation levels or lengths of the heavy chain complementarity-determining region 3. Since RSV repeatedly infects every person throughout life, memory B cells (MBC) expressing RSV F-binding BCRs circulate in the blood of healthy adults. This dataset of BCR variable region sequence features was derived from single cell-sorted RSV F-directed MBCs of a healthy adult blood donor [1]. The dataset was produced with publicly available data analysis software programs and scripts, which facilitates integration or comparison with antibody sequence repertoire data of different individuals derived with the same or comparable data analysis approaches and tools.


Brain content of immunoreactive [Leu5]enkephalin and [Met5]enkephalin after pentylenetetrazol-induced convulsions.
Wistar rats were injected daily with convulsant doses of PTZ (40 mg/kg). The brain content of immunoreactive (IR) enkephalins was measured after repetitive clonic convulsions and after the 'kindling' state was reached. In both instances we found a significant increase of IR [Leu5]enkephalin and IR [Met5]enkephalin in septum, striatum and amygdala 24 h after PTZ administration. These results support the suggestion that the peptide increase occurs with repetitive convulsions and not only after the 'kindling' state is reached.


Effectiveness of the WeChat-based smoking cessation intervention ('WeChat WeQuit' program) in China: study protocol for a randomized controlled trial.
Since its first release in 2011, WeChat (Chinese pinyin: Wei Xin) has been widely used to promote health. As the most popular all-in-one social media application providing free instant messaging, voice, text, pictures, videos and location information exchange via smartphone, WeChat holds promise for delivering smoking cessation services to a large number of people at low unit cost in China, a country with more than 300 million smokers. This protocol is for a study that aims to assess the efficacy of a WeChat-based smoking cessation intervention ('WeChat WeQuit') for treatment-seeking smokers in China. In this two-arm randomized controlled trial (RCT), the control group will receive only a digital version of a booklet of 'Happy Quit'. The intervention group will also receive the 2 weeks pre- and 12 weeks post-quit 'WeChat WeQuit' intervention. Both groups will be followed-up to 26 weeks (6 months) after quit day. This on-line trial has no restrictions on setting or location. Chinese-speaking current smokers (have smoked ≥100 cigarettes in their life-time and currently smoke cigarettes) aged 18+ years willing to make a quit attempt within 1 month (n = 2000) will be recruited by on- and off-line advertising. Potential participants will register their interest by sending WeChat messages or text messages, or making a voice call. The primary outcome measure is self-reported 26-week continuous smoking abstinence with biological verification [test by clinic visit: expired carbon monoxide concentration <10 parts per million (p.p.m.) or by mailing cotinine urine dipsticks: urine cotinine <200 ng/ml]. Secondary outcome measures include: 7-day point prevalence abstinence at weeks 1, 4, 8, 12, 16, 20 and 26 after-quit day; self-reported 12-week continuous smoking abstinence and 26-week continuous smoking abstinence; reductions in number of cigarettes smoked per day; and rates of participation in and completion of the smoking-cessation program. If the 'WeChat WeQuit' program is found to be effective, it will be an accessible, affordable and widely implementable intervention to help cigarette smokers to quit. The potential public health impact of its expansion into underserved regions or communities is considerable.


Undifferentiated sebaceous carcinoma: an unusual childhood cancer.
Sebaceous carcinoma is an aggressive, adnexal, rare malignant tumor that may arise in ocular or extra-ocular sites. Extraorbital sebaceous carcinoma is exceptional in childhood. We report a 12-year-old boy with an ocular sebaceous carcinoma who was first seen with an asymptomatic firm, cutaneous nodule on the right eyebrow. The tumor developed slowly within 1 year. Histologically, it was an undifferentiated sebaceous carcinoma. The patient had surgery with wide surgical margins. He was alive and free from disease at a follow-up of 34 months. Close follow-up of this tumor is recommended because of the risk of aggressive behavior.


Motionless volumetric photoacoustic microscopy with spatially invariant resolution.
Photoacoustic microscopy (PAM) is uniquely positioned for biomedical applications because of its ability to visualize optical absorption contrast in vivo in three dimensions. Here we propose motionless volumetric spatially invariant resolution photoacoustic microscopy (SIR-PAM). To realize motionless volumetric imaging, SIR-PAM combines two-dimensional Fourier-spectrum optical excitation with single-element depth-resolved photoacoustic detection. To achieve spatially invariant lateral resolution, propagation-invariant sinusoidal fringes are generated by a digital micromirror device. Further, SIR-PAM achieves 1.5 times finer lateral resolution than conventional PAM. The superior performance was demonstrated in imaging both inanimate objects and animals in vivo with a resolution-invariant axial range of 1.8 mm, 33 times the depth of field of the conventional PAM counterpart. Our work opens new perspectives for PAM in biomedical sciences.Photoacoustic microscopy allows for label-free 3D in vivo imaging by detecting the acoustic response of a photoexcited material. Here, Yang et. al use a digital-micromirror-device based structured illumination scheme to both improve resolution and greatly increase the depth of field, enabling 3D volumetric imaging.


Perception of causality and synchrony dissociate in the audiovisual bounce-inducing effect (ABE).
A sound can cause 2 visual streaming objects appear to bounce (the audiovisual bounce-inducing effect, ABE). Here we examined whether the stream/bounce percept affects perception of audiovisual synchrony. Participants saw 2 disks that either clearly streamed, clearly bounced, or were ambiguous, and heard a sound around the point of contact (POC). They reported, on each trial, whether they perceived the disks to 'stream' or 'bounce', and whether the sound was 'synchronous' or 'asynchronous' with the POC. Results showed that the optimal time of the sound to induce a bounce was before the POC (-59 msec), whereas audiovisual synchrony was maximal when the sound came after the POC (+16 msec). The range of temporal asynchronies perceived as 'synchronous', the temporal binding window (TBW), was wider when disks were perceived as bouncing than streaming, with no difference between ambiguous and non-ambiguous visual displays. These results demonstrate 1) that causality differs from synchrony, 2) that causality widens the TBW, and 3) that the ABE is perceptually real.


Retracted: The Dissemination, Influence, and Efficiency of Jameson's Cultural Theory Combined with Chinese Mass Culture and Mass Sports Culture in the Sustainable Development of China.
[This retracts the article DOI: 10.1155/2022/6510147.].


Conformational analysis of NMDA receptor GluN1, GluN2, and GluN3 ligand-binding domains reveals subtype-specific characteristics.
The NMDA receptor family of glutamate receptor ion channels is formed by obligate heteromeric assemblies of GluN1, GluN2, and GluN3 subunits. GluN1 and GluN3 bind glycine, whereas GluN2 binds glutamate. Crystal structures of the GluN1 and GluN3A ligand-binding domains (LBDs) in their apo states unexpectedly reveal open- and closed-cleft conformations, respectively, with water molecules filling the binding pockets. Computed conformational free energy landscapes for GluN1, GluN2A, and GluN3A LBDs reveal that the apo-state LBDs sample closed-cleft conformations, suggesting that their agonists bind via a conformational selection mechanism. By contrast, free energy landscapes for the AMPA receptor GluA2 LBD suggest binding of glutamate via an induced-fit mechanism. Principal component analysis reveals a rich spectrum of hinge bending, rocking, twisting, and sweeping motions that are different for the GluN1, GluN2A, GluN3A, and GluA2 LBDs. This variation highlights the structural complexity of signaling by glutamate receptor ion channels.


The Prognostic Value of Qualitative and Quantitative Stress CMR in Patients with Known or Suspected CAD.
Recent studies suggest that quantitative cardiac magnetic resonance (CMR) may have more accuracy than qualitative CMR in coronary artery disease (CAD) diagnosis. However, the prognostic value of quantitative and qualitative CMR has not been compared systematically. The objective was to conduct a systematic review and meta-analysis assessing the utility of qualitative and quantitative stress CMR in the prognosis of patients with known or suspected CAD. A comprehensive search was performed through Embase, Scopus, Web of Science, and Medline. Studies that used qualitative vasodilator CMR or quantitative CMR assessments to compare the prognosis of patients with positive and negative CMR results were extracted. A meta-analysis was then performed to assess: 1) major adverse cardiovascular events (MACE) including cardiac death, nonfatal myocardial infarction (MI), unstable angina, and coronary revascularization; and 2) cardiac hard events defined as the composite of cardiac death and nonfatal MI. Forty-one studies with 38,030 patients were included in this systematic review. MACE occurred significantly more in patients with positive qualitative (HR: 3.86; 95% CI: 3.28-4.54) and quantitative (HR: 4.60; 95% CI: 1.60-13.21) CMR assessments. There was no significant difference between qualitative and quantitative CMR assessments in predicting MACE (P = 0.75). In studies with qualitative CMR assessment, cardiac hard events (OR: 7.21; 95% CI: 4.99-10.41), cardiac death (OR: 5.63; 95% CI: 2.46-12.92), nonfatal MI (OR: 7.46; 95% CI: 3.49-15.96), coronary revascularization (OR: 6.34; 95% CI: 3.42-1.75), and all-cause mortality (HR: 1.66; 95% CI: 1.12-2.47) were higher in patients with positive CMR. The presence of myocardial ischemia on CMR is associated with worse clinical outcomes in patients with known or suspected CAD. Both qualitative and quantitative stress CMR assessments are helpful tools for predicting clinical outcomes.


The synaptic localization of NR2B-containing NMDA receptors is controlled by interactions with PDZ proteins and AP-2.
The NMDA receptor (NMDAR) is a component of excitatory synapses and a key participant in synaptic plasticity. We investigated the role of two domains in the C terminus of the NR2B subunit--the PDZ binding domain and the clathrin adaptor protein (AP-2) binding motif--in the synaptic localization of NMDA receptors. NR2B subunits lacking functional PDZ binding are excluded from the synapse. Mutations in the AP-2 binding motif, YEKL, significantly increase the number of synaptic receptors and allow the synaptic localization of NR2B subunits lacking PDZ binding. Peptides corresponding to YEKL increase the synaptic response within minutes. In contrast, the NR2A subunit localizes to the synapse in the absence of PDZ binding and is not altered by mutations in its motif corresponding to YEKL of NR2B. This study identifies a dynamic regulation of synaptic NR2B-containing NMDARs through PDZ protein-mediated stabilization and AP-2-mediated internalization that is modulated by phosphorylation by Fyn kinase.


Depletion of platelet amine storage granules by the antithrombotic agent, suloctidil.
Suloctidil, a compound with antithrombotic properties in animal models, causes depletion of human platelet serotonin stores during 1-5 hr incubation with platelet-rich plasma in vitro. This effect is not attended by leakage of cytoplasmic nucleotides or by alterations in cyclic AMP metabolism, malondialdehyde production or energy balance. 5HT uptake was also inhibited but uptake of adenine was not. 5HT released by suloctidil appeared in the supernatant as oxidation products, though the amine also accumulated when re-uptake was blocked by imipramine.


[Study of blending method for the extracts of herbal plants].
The irregularity in herbal plant composition is influenced by multiple factors. As for quality control of traditional Chinese medicine, the most critical challenge is to ensure the dosage content uniformity. This content uniformity can be improved by blending different batches of the extracts of herbal plants. Nonlinear least-squares regression was used to calculate the blending coefficient, which means no great absolute differences allowed for all ingredients. For traditional Chinese medicines, even relatively smaller differences could present to be very important for all the ingredients. The auto-scaling pretreatment was used prior to the calculation of the blending coefficients. The pretreatment buffered the characteristics of individual data for the ingredients in different batches, so an improved auto-scaling pretreatment method was proposed. With the improved auto-scaling pretreatment, the relative. differences decreased after blending different batches of extracts of herbal plants according to the reference samples. And the content uniformity control of the specific ingredients could be achieved by the error control coefficient. In the studies for the extracts of fructus gardeniae, the relative differences of all the ingredients is less than 3% after blending different batches of the extracts. The results showed that nonlinear least-squares regression can be used to calculate the blending coefficient of the herbal plant extracts.


Progression of carotid atherosclerosis in patients with systemic lupus erythematosus.
The progression of carotid atherosclerosis in lupus patients is frequently encountered, and it is determined by both traditional and nontraditional risk factors. Of the 181 patients initially included in the study, 157 patients were reevaluated after 39(37-42) months. The progression of atherosclerosis was defined as the increase in the intima-media thickness (IMT) >0.15 mm and/or an increase of the plaque score. The predictive factors of progression were identified using the Poisson regression model. The median of the cohort age at baseline was 38 years (range 29-46 years; 96.2% female, 75.8% nonwhite). Carotid atherosclerosis progression was observed in 43 patients (27.4%), an increased plaque score was observed in nine patients (5.7%), an increase of IMT >0.15 mm was observed in 31 (19.7%), and both issues were present in three patients (1.9%). The univariate determinants of atherosclerosis progression were age, systemic lupus erythematosus (SLE) duration, and higher serum level of triglycerides (p < 0.05). The presence of nephrotic proteinuria (p = 0.063), stage 3 or greater chronic kidney disease (p = 0.091), and longer duration of prednisone use (p = 0.056) showed a tendency towards association with progression of atherosclerosis. The independent risk factors for progression were the SLE duration (p = 0.008, RR = 1.06, 95% CI = 1.03-1.10) and the presence of nephrotic proteinuria (p = 0.022, RR = 4.22, 95% CI = 2.18-8.15). The progression of atherosclerosis occurred in a substantial number of young SLE patients during a short-term follow-up. The independent factors associated with this progression emphasize the importance of SLE in determining atherosclerosis in these individuals.


Deformation behavior and properties of severe plastic deformation techniques for bulk materials: A review.
This paper presents a comprehensive review of the successful application of Severe Plastic Deformation (SPD) in producing ultra-fine grained (UFG) and nano-structured crystalline bulk materials. SPD achieves outstanding grain refinement without significantly altering the original dimensions of the workpiece, making it particularly useful for ductile materials that can withstand large strains under high hydrostatic pressure before failure. The study explores the grain refining mechanism during severe plastic deformation and its impact on the microstructure of metals. It also examines the use of SPD in hard to deform brittle materials like tungsten oxide, B2O3 glasses, and amorphous materials. The paper discusses the advantages and disadvantages of each technique, along with their applications and potential for combining more than one technique. The review is significant because it emphasizes recent progress in process development, which could potentially enable the industrialization of certain SPD techniques for specific applications. This paper fills the gap in the literature by addressing this issue. Overall, the review demonstrates the potential of SPD in metalworking and its application in the development of new UFG materials with improved mechanical properties.


[Long-term results in the surgical treatment of pilon tibial fractures. A retrospective study].
While Pilon fractures of the tibia have been treated for decades by primary open reduction and internal fixation by plate osteosynthesis, during the last 10 years differential treatment was developed: After primary open reduction nowadays patients are treated with (according to type of fracture and tissue damage). As well as primary open reduction and internal fixation a two-step treatment (primary external fixator and delayed ORIF) or consolidation by external fixator combined with minimal invasive osteosynthesis (cannulated screws and K-wires) has been implemented. Furthermore, the significance of primary bone grafting in comminuted fractures to prevent aseptic pseudarthrosis has been acknowledged. Of 151 patients with 160 pilon fractures treated from January 1979 to May 1995, 107 patients (113 fractures) were evaluated. Only the results of C2 and C3 fractures could be compared, as only in these groups were all three types of treatment used. Over 75% of the treated fractures were closed fractures, most of them being fractures with a soft tissue damage grade 2 of the Oestern and Tscherne classification. In the open fractures we found mainly grade 3 fractures according to the Gustilo and Anderson classification. In 54.9% of all pilon tibial fractures we observed an uncomplicated course of healing. Early complications (25.7%) were mainly soft tissue infections, whereas we found pseudarthrosis to be the most frequent late complication. Highest infection rate (55.5%) was in the two-step treatment group (primary external fixator and delayed ORIF) and lowest in the primary internal stabilization group, although especially in the C2 and C3 fractures best clinical late results were obtained with the two-step procedure. The complication rate in the treatment of pilon fractures depends mainly on the type of fracture, the soft tissue damage and the type of treatment. The results of primary ORIF varied. In the case of low-grade soft tissue damage, good to excellent results were accomplished. In the case of higher-grade soft tissue damage, the problem of soft tissue coverage and reconstruction of the joint surface could be solved with good results by the two-step treatment. Herewith it is important to use limited open reduction of displaced fragments and fixation by cannulated screws and K-wires. We consider ORIF of the fibula necessary as stabilization of the second column of the ankle joint.


Monoclonal antibodies detect polymorphic determinants shared by I-A and I-E antigens.
Binding data on inbred mouse strains and immunochemical isolation of Ia antigens with subsequent separation on non-reduced/reduced two-dimensional gels provide evidence for the cross-reactivity of monoclonal antibodies with I-A and I-E products. Thus two monoclonal antibodies were found to react with A alpha A beta as well as E alpha E beta dimers. One of these mAbs, K22 -42, reacts with the precursor form of E beta chain of B10.GD mice which is associated with the invariant chain (Ii). This indicates that the respective determinant on E beta is formed prior to association of E beta with E alpha.


[Trends in smoking among adults in Germany : Evidence from seven population-based health surveys from 1991-2015].
Despite decreasing smoking prevalence, tobacco use remains a key public health problem in Germany. For planning, managing, and evaluating tobacco control measures, regular data collection on smoking behavior in the population is essential. The aim of this article is to present trends in adult tobacco use since the early 1990s based on data from the health monitoring of the Robert Koch-Institute (RKI). Analyses are based on data from 106,158 individuals aged 18 to 79 years, who participated in seven RKI health surveys from 1991-2015. Trends in tobacco consumption were analyzed using different indicators of smoking behavior, stratified by age, cohort, and gender. An overall falling smoking prevalence can be attributed primarily to a significant decline in the younger age groups since the early 2000s. Trend analysis by cohort reveals a declining prevalence for almost all cohorts over time from 1991-2015. Historically there has been is a sharp increase in the prevalence of women who have ever smoked between the 1930-1934 and 1950-1959 cohorts. The proportion of men who have ever smoked slightly decreased between the 1930-1934 and 1980-1984 cohorts. The shown overall decline happened concurrently with various tobacco prevention measures implemented during this period in Germany. If present trends related to continuous high smoking rates are sustained, it can be assumed that the tobacco consumption of the population will remain the source of adverse health outcomes. Accordingly, tobacco prevention measures and the promotion of smoking cessation in all age groups should be a public health priority.


Transcriptome analysis reveals immune regulation in the spleen of koi carp (Cyprinus carpio Koi) during Aeromonas hydrophila infection.
A. hydrophila (Aeromonas hydrophila) is one of the most hazardous pathogenic microorganisms threatening the aquaculture industry and exhibits zoonotic-like characteristics. This study was designed to investigate the differential gene expression and pathway enrichment in the spleen of koi carp (Cyprinus carpio koi) upon A. hydrophila infection. The Illumina NovaSeq 6000 sequencing platform was used to identify 252 DEGs (differentially expressed genes), including 112 upregulated genes and 140 downregulated genes, in the spleens of koi carp challenged with A. hydrophila compared to those in the spleens of koi carp treated with PBS (phosphate-buffered saline). DEGs were shown to be involved in 133 pathways by KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis. Numerous immunological disease-related pathways, such as the immune defense network for IgA production, Staphylococcus aureus infection, and antigen processing and presentation, were enriched in the DEGs. In addition, the expression levels of 10 randomly screened DEGs, including the inflammatory factor nlrp3 (NOD-like receptor family pyrin domain containing 3), cytokine il-8 (interleukin-8), c2 (complement c2), c3 (complement c3), and the lipid mediator cox1 (cyclooxygenase-1), were compared by qPCR. The results showed that six genes, including il-8, cox1, and nlrp3, were upregulated according to both RNA-seq and qPCR validation, while four, including c2 and c3, showed downregulated expression. This result verified a strong correlation between the RNA-seq and qPCR datasets at the expression level. Moreover, this study provided splenic transcriptome data for koi carp during A. hydrophila infection and provided theoretical support for future drug development.


Nitrogen dioxide reducing ascorbic acid technologies in the ventilator circuit leads to uniform NO concentration during inspiration.
Conventional inhaled NO systems deliver NO by synchronized injection or continuous NO flow in the ventilator circuitry. Such methods can lead to variable concentrations during inspiration that may differ from desired dosing. NO concentrations in these systems are generally monitored through electrochemical methods that are too slow to capture this nuance and potential dosing error. A novel technology that reduces NO2 into NO via low-resistance ascorbic-acid cartridges just prior to inhalation has recently been described. The gas volume of these cartridges may enhance gas mixing and reduce dosing inconsistency throughout inhalation. The impact of the ascorbic-acid cartridge technology on NO concentration during inspiration was characterized through rapid chemiluminescence detection during volume control ventilation, pressure control ventilation, synchronized intermittent mandatory ventilation and continuous positive airway pressure using an in vitro lung model configured to simulate the complete uptake of NO. Two ascorbic acid cartridges in series provided uniform and consistent dosing during inspiration during all modes of ventilation. The use of one cartridge showed variable inspiratory concentration of NO at the largest tidal volumes, whereas the use of no ascorbic acid cartridge led to highly inconsistent NO inspiratory waveforms. The use of ascorbic acid cartridges also decreased breath-to-breath variation in SIMV and CPAP ventilation. The ascorbic-acid cartridges, which are designed to convert NO2 (either as substrate or resulting from NO oxidation during injection) into NO, also provide the benefit of minimizing the variation of inhaled NO concentration during inspiration. It is expected that the implementation of this method will lead to more consistent and predictable dosing.


Reducing the hindsight bias utilizing attorney closing arguments.
In the legal system, jurors are asked to render a decision after the event in question has already occurred and the final outcome, typically negative, is known. This "after-the-fact" structure of the legal system makes jurors susceptible to a human judgment phenomenon known as hindsight bias. This study focused on reducing hindsight bias in a courtroom context by incorporating a debiasing strategy within the defense's closing argument. Subjects viewed one of three videotaped versions of plaintiff and defense closing arguments in a commercial litigation case (i.e., foresight condition, hindsight condition, and hindsight debiasing condition). Results indicate that the hindsight debiasing strategy was effective in reducing subject-juror hindsight bias.


Induction of multiple replacement mutations by oligonucleotide-directed mutagenesis with extended mismatch primers.
We have developed a method called oligo-scanning mutagenesis that uses oligonucleotides to mutate up to 12 contiguous bases in a single step. Some advantages of this procedure are that the position and sequence of the replacement mutations are completely specified by the investigator, and combinations of mutations can easily be generated. The technique uses a gapped substrate and the Escherichia coli dam methylation error-correcting mechanism to increase the yield of mutants.


Assessment of the acute effects of glucocorticoid treatment on coronary microembolization using cine, first-pass perfusion, and delayed enhancement MRI.
To assess the acute effects of methylprednisone treatment (MPT) on coronary microembolization (CME) by cardiac cine, first-pass perfusion, and delayed gadolinium enhancement magnetic resonance imaging (DE-MRI) in an experimental swine model. Microembolization was established by intracoronary infusion of microspheres into the left anterior artery. Swine received placebo (n = 12) or methylprednisolone (n = 10, 30 mg/kg) intravenously 30 minutes before microembolization. Perfusion and DE-MRI was performed 6 hours after microembolization. Cine MR images of pre-/post-CME were obtained using 1.5T scanner. Cine MRI demonstrated relative amelioration of the post-CME myocardial contractile dysfunction in the glucocorticoid-treated group compared to the placebo group (P < 0.001). Post-CME target myocardial perfusion parameters decreased in both groups after microembolization. The extent of these decreases were the same for the embolized-to-control area ratio of maximum upslope (P = 0.245; 95% confidence interval of the difference [CID], -0.041/0.148) and time to peak ratio (P = 0.122; 95% CID, -0.201/0.026); however, the maximum signal intensity was higher in the glucocorticoid-treated group (P = 0.012; 95% CID, 0.023/0.156). DE-MRI revealed patchy hyperenhancement in all placebo pigs (n = 12/12) after microembolization, but no hyperenhanced regions in the glucocorticoid-pretreated pigs (n = 0/10). Standard, readily available, cardiac MRI techniques are useful in demonstrating post-CME myocardial dysfunction and the acute effects of glucocorticoid treatment on CME. Glucocorticoid pretreatment improves myocardial contractile dysfunction, prevents hyperenhancement, and partially ameliorates the decline of myocardial perfusion in the embolized area.


The dynamics of the emergency medical readmission - The underlying fundamentals.
Hospital readmissions are a perennial problem. We reviewed readmissions to one institution (2002-2015) and investigated their dynamics. 96,474 emergency admissions (in 50,701 patients) to an Irish hospital over a 15-year period were studied, and patterns surrounding early (<28days) and late (any other) readmissions determined. Univariate and logistic or truncated Poisson regression methods were employed. Early readmission rate averaged 9.6% (95% CI: 9.4, 9.8) with a low/high of 8.4% (95% CI: 7.8, 9.1) and 10.3% (95% CI: 9.6, 11.0) respectively with no overall time trend. Early readmissions represented 20.1% (95% CI: 19.8, 20.5) of emergency medical readmissions. Median time to first readmission was 55weeks (95% CI: 13, 159), time to second was 35weeks (95% CI: 9, 98); by the 7th/8th readmissions, intervals were 13weeks (95% CI: 4, 36) and 11weeks (95% CI: 4, 30). Readmissions were older 67.1years (95% CI: 48.3, 79.2) vs. single admissions 53.9years (34.3, 72.4) and stayed longer - 5.8days (2.7, 10.6) vs. 3.9days (1.5, 8.0). Readmissions had more Acute Illness Severity, Charlson Co-Morbidity and Chronic Disabling Disease. Between 2002 and 2015 the logistic adjusted model of 30-day in-hospital mortality reduced from 6.1% (95% CI: 5.7, 6.5) to 4.4% (95% CI: 4.1, 4.7) (RRR 30.4%). Early hospital readmission rate did not change over 15years despite improvements in hospital mortality outcomes. Readmissions have a consistent pattern related to patient illness and social characteristics; the fundamentals are driven by disease progression over time.