Biotin-transfer from a trifunctional crosslinker for identification of cell surface receptors of soluble protein ligands.
Here we describe a novel crosslinker and its application as a biotin-transfer reagent to identify cell surface receptors of soluble protein ligands on live cells. This crosslinker contains three functional groups: an aldehyde-reactive aminooxy group, a sulfhydryl, and a biotin (ASB). It is readily synthesized via a 3-step addition reaction using standard solid-phase peptide synthesis methods and commercially available intermediates, allowing access to laboratories without specialized synthetic chemistry capabilities. For the biotin-transfer method, ASB is linked to a protein ligand through the sulfhydryl group in a two-step process that allows the introduction of a disulfide bond between the ligand and the crosslinker. Incubation of the labelled ligand with oxidized live cells leads to the formation of crosslinks with aldehyde-containing glycans on the cell surface receptor. Subsequent reduction of the disulfide bond results in biotin transfer from the ligand to the cell surface receptor. Protein biotinylation that is mediated by ligand binding to its receptor is differentiated from background biotinylation events by comparison with a similarly labelled control protein using comparative proteomic mass spectrometry to quantify streptavidin-bound proteins. Using this method, we successfully identified the cell surface receptors of a peptide hormone, a monoclonal antibody, and a single-domain antibody-Fc fusion construct.


Port-access approach for combined aortic and mitral valve surgery.
A technique is described for combined aortic and mitral valve operation utilizing the minimally invasive Port-Access technique. Two patients are repaired with excellent chest wall healing and avoidance of sternotomy.


Chronic impairment of mitochondrial bioenergetics and β-oxidation promotes experimental AKI-to-CKD transition induced by folic acid.
Recent studies suggest that mitochondrial bioenergetics and oxidative stress alterations may be common mechanisms involved in the progression of renal damage. However, the evolution of the mitochondrial alterations over time and the possible effects that their prevention could have in the progression of renal damage are not clear. Folic acid (FA)-induced kidney damage is a widely used experimental model to induce acute kidney injury (AKI), which can evolve to chronic kidney disease (CKD). Therefore, it has been extensively applied to study the mechanisms involved in AKI-to-CKD transition. We previously demonstrated that one day after FA administration, N-acetyl-cysteine (NAC) pre-administration prevented the development of AKI induced by FA. Such therapeutic effect was related to mitochondrial preservation. In the present study, we characterized the temporal course of mitochondrial bioenergetics and redox state alterations along the progression of renal damage induced by FA. Mitochondrial function was studied at different time points and showed a sustained impairment in oxidative phosphorylation capacity and a decrease in β-oxidation, decoupling, mitochondrial membrane potential depolarization and a pro-oxidative state, attributed to the reduction in activity of complexes I and III and mitochondrial cristae effacement, thus favoring the transition from AKI to CKD. Furthermore, the mitochondrial protection by NAC administration before AKI prevented not only the long-term deterioration of mitochondrial function at the chronic stage, but also CKD development. Taken together, our results support the idea that the prevention of mitochondrial dysfunction during an AKI event can be a useful strategy to prevent the transition to CKD.


Improvement of the maxillary bone growth suppression in the cleft palate operation with cultured dermal substitute: animal experiment and patient reports in preliminary clinical application.
Cleft palate patients often show impaired maxillary bone growth after cleft-palate-correction surgery. We attempted to investigate and elucidate the effects of using allogeneic, cultured dermal substitute (CDS) to cover an exposed, palatal bone surface in animal experiments. Fibroblasts from the abdominal skin of Wistar rats were cultured. Subsequently, the fibroblasts were seeded onto a matrix that composed of hyaluronic acid and atelo-collagen. Forty Wistar rats (3-week-old males) were assigned to one of four groups: control, open-treatment, matrix and CDS groups. The control group (n=5) received no surgical operations. In the open-treatment group (n=11), the mucosa and periosteum of the left-half of the palate were removed surgically and the bone was exposed. In the matrix group (n=11), the area of exposed bone was covered with only the matrix, excluding any cells. In the CDS group (n=10), the area of exposed bone was covered with CDS. At 9 weeks postoperatively, biopsies of the wounds were obtained. Skull preparations were made and the palatal widths were determined. The palatal widths in the CDS group were significantly wider compared to the matrix and open-treatment groups (P<0.05). However, there were no significant differences when the CDS group was compared to the control group. Haematoxylin, eosin and CD31 immunostaining confirmed a larger number of capillaries in the CDS group. This animal experiment suggested that this procedure might provide an optimum wound-healing condition, thus, reducing the maxillary bone-growth suppression. Therefore, a preliminary clinical application in three patients was performed using the autologous CDS after the pushback method.


Inducing Hetero-aggregation of Different Azo Dyes through Electrostatic Self-Assembly.
Combining chemically different building blocks in supramolecular nanoparticles is a promising key to tailored structures and functionalities. π-π heterostacks of dye molecules form upon electrostatic self-assembly with a polyelectrolyte, resulting in stable ternary nano-assemblies in aqueous solution. Core-shell spheres, cylinders, and flexible cylinders result, which exhibit new shapes different from the binary systems. Particle shapes can be tuned through the dye composition.


Structural basis for the recognition of Asef by adenomatous polyposis coli.
Adenomatous polyposis coli (APC) regulates cell-cell adhesion and cell migration through activating the APC-stimulated guanine nucleotide-exchange factor (GEF; Asef), which is usually autoinhibited through the binding between its Src homology 3 (SH3) and Dbl homology (DH) domains. The APC-activated Asef stimulates the small GTPase Cdc42, which leads to decreased cell-cell adherence and enhanced cell migration. In colorectal cancers, truncated APC constitutively activates Asef and promotes cancer cell migration and angiogenesis. Here, we report crystal structures of the human APC/Asef complex. We find that the armadillo repeat domain of APC uses a highly conserved surface groove to recognize the APC-binding region (ABR) of Asef, conformation of which changes dramatically upon binding to APC. Key residues on APC and Asef for the complex formation were mutated and their importance was demonstrated by binding and activity assays. Structural superimposition of the APC/Asef complex with autoinhibited Asef suggests that the binding between APC and Asef might create a steric clash between Asef-DH domain and APC, which possibly leads to a conformational change in Asef that stimulates its GEF activity. Our structures thus elucidate the molecular mechanism of Asef recognition by APC, as well as provide a potential target for pharmaceutical intervention against cancers.


Preventive Impella® Support in High-Risk Patients Undergoing Cardiac Surgery.
Patients with severely reduced LV-EF ≤ 30% undergoing CABG have a high risk for postoperative cardiogenic shock. The optimal timing of an adequate hemodynamic support has an impact on short- and midterm mortality after CABG. This study aimed to assess the prophylactic use of the Impella pump in high-risk patients undergoing elective cardiac surgery. In this single-center retrospective study, 14 patients with LV-EF (≤30%) undergoing cardiac surgery received a prophylactic, perioperative Impella (5.0, 5.5) support between 2020 and 2022. The mean age at surgery was 64.2 ± 2.6 years, the mean preoperative LV-EF was 20.7% ± 1.56%. The duration of Impella support was 4 (3-7.8) days and the 30-day survival rate was 92.85%. Acute renal failure occurred in four patients who were dialyzed on average for 1.2 ± 0.7 days. Mechanical ventilation was needed for 1.75 (0.9-2.7) days. Time to inotrope/vasopressor independence was 2 (0.97-7.25) days with a highest lactate level (24 h postoperatively) of 3.8 ± 0.6 mmol/l. Postoperative LV-EF showed a significant improvement when compared to preoperative LV-EF (29.1% ± 2.6% vs. 20.7% ± 1.56% (p = 0.022)). The prophylactic Impella application seems to be a safe approach to improve the outcomes of this patient population.


Pegylated Liposomal Doxorubicin/Oxaliplatin Chemotherapy Can Overcome Cisplatin Resistance in Spectrin αII-Overexpressing Ovarian Carcinoma.
Spectrin αII contributes to cisplatin and carboplatin resistance in ovarian serous carcinoma cells, and its expression in surgical specimens is a valid predictor of prognosis. We sought to identify effective drugs for spectrin αII-mediated cisplatin-resistant cells. We employed SKOV3 cells with small interfering RNA-mediated spectrin αII downregulation, serous carcinoma cells (NOS2), cisplatin-resistant cells (NOS2CR2), and oxaliplatin-resistant cells (NOS2OXR). In the drug-sensitivity test, oxaliplatin was not affected by the inhibition of spectrin αII expression and was effective for cisplatin-resistant NOS2CR2 cells. NOS2OXR cells did not express higher levels of spectrin αII compared to NOS2 in western blot analysis. Six non-platinum anticancer drugs were not affected by the inhibition and was effective for resistant NOS2CR2 and NOS2OXR cells. Doxorubicin exhibited potent cytotoxicity at 2 μM against both resistant cell lines. Pegylated liposomal doxorubicin/oxaliplatin regimen may be effective for cisplatin-resistant ovarian carcinoma with spectrin αII-overexpression.


Prevalence of Helicobacter pylori antibodies in children in Bloemfontein, South Africa.
An association of H. pylori infection with chronic gastritis, peptic ulceration and gastric cancer is known. Prevalence of IgG antibodies to Helicobacter pylori in children in the Bloemfontein, South Africa area was studied. Children attending the general pediatric outpatient department at Pelonomi Hospital in Bloemfontein were grouped according to age. A minimum of 100 children was investigated in each age group. Baseline demographic and socioeconomic data were collected. The study showed a high prevalence of H. pylori antibodies. Prevalence increased with age: 13.5% in children 3 months-2 years, 48.5% at 2-5 years, 67.3% at 5-10 years and 84.2% at 10-15 years. Investigation of the socioeconomic data in relation to the prevalence of H. pylori was inconclusive. This high prevalence needs further study.


In vitro antibacterial activity and durability of a nano-curcumin-containing pulp capping agent combined with antimicrobial photodynamic therapy.
Considering the antibacterial properties of nano-curcumin (nCur) reinforced with antimicrobial photodynamic therapy (aPDT), this study aimed to assess the antibacterial activity and durability of Activa BioActive Base/Liner (ABBL) containing nCur (nCur-ABBL) as a pulp capping agent against Streptococcus mutans, the most common cause of secondary caries. In this in vitro experimental study, ABBL discs containing 0.5 %, 1%, 2%, and 5% (w/w) concentrations of nCur were fabricated. After aPDT using light emitting diode (LED) at 435 ± 20 nm wavelength for 5 min, the discs were undergone aging in artificial saliva for 90 days. The antibacterial activity of the discs against S. mutans was evaluated by the disc agar diffusion test, and the number of bacterial colonies present in the biofilm formed on the disc surfaces was counted after 0, 15, 30, and 60 days of aging. The maximum growth inhibition zone was noted around the 5% nCur-ABBL discs. Increasing the concentration of nCur from 0.5 % to 5% combined with aPDT significantly decreased the number of S. mutans colonies in the biofilm over time (P < 0.05). nCur-ABBL discs containing 2% and 5% nCur had no difference in antibacterial activity at any time point up to 60 days (P > 0.05). According to our data, 5% nCur-ABBL revealed the largest growth inhibition zone in S. mutans culture. Moreover, 5% nCur can serve as an excellent ABBL additive in aPDT producer against S. mutans biofilms up to 60 days of aging period.


Koeleria macrantha: performance and distribution in relation to soil and plant calcium and magnesium.
• The reported sensitivity of Koeleria macrantha (Poaceae) to soil magnesium, resulting in its absence from South Wales dolomitic limestone, was examined here in relation to varying ionic Ca to Mg ratios, and by cultivation in different limestone soils. • In a growth cabinet experiment, shoot and root Ca and Mg concentration and dry weight yield were determined for five edaphically varied populations of K. macrantha grown from tillers, over a range of Ca : Mg concentrations. The influence of relative concentrations of other nutrients was also investigated. In addition, K. macrantha plants were cultivated on Carboniferous, Magnesian and dolomitic limestone soils. • Total plant Ca: Mg ranged from 0.3 to > 20 mille-equivalents. Optimal substrate ratios (from 25 : 1 to 0.1 : 1), and response to different concentrations of nutrients varied between the populations. Cultivation on dolomitic limestone soil produced the highest yields. • The adverse effect on yields of all populations with low substrate Ca: Mg was much lower than predicted. Results suggest K. macrantha has a higher substrate Mg tolerance than other members of the Poaceae: its reported absence from the South Wales dolomitic limestone is unlikely to be due to soil magnesium sensitivity.


Room-temperature chemical synthesis of C2.
Diatomic carbon (C2) is historically an elusive chemical species. It has long been believed that the generation of C2 requires extremely high physical energy, such as an electric carbon arc or multiple photon excitation, and so it has been the general consensus that the inherent nature of C2 in the ground state is experimentally inaccessible. Here, we present the chemical synthesis of C2 from a hypervalent alkynyl-λ3-iodane in a flask at room temperature or below, providing experimental evidence to support theoretical predictions that C2 has a singlet biradical character with a quadruple bond, thus settling a long-standing controversy between experimental and theoretical chemists, and that C2 serves as a molecular element in the bottom-up chemical synthesis of nanocarbons such as graphite, carbon nanotubes, and C60.


Clastogenic, anti-clastogenic profile and safety assessment of Camel urine towards the development of new drug target.
Camel Urine (CU) is composed of components that have antitumor properties and other therapeutic benefits. Regardless of short-term preliminary CU genotoxicity is reported, comprehensive genotoxic studies are limited. In this study, sensitive in vitro and in vivo genotoxic bioassays such as mitotic index (MI), chromosomal aberrations (CA), micronucleated polychromatic erythrocytes (MPE), and analysis of primary spermatocytes were employed. The adventitious roots of Allium cepa L. and mice (Mus musculus), as an experimental mammalian system, were employed to assess the MI and CA of CU induced by sodium nitrate and cyclophosphamide respectively. In contrast, other clastogenic assays were studied in mice (Mus musculus). Twenty-eight days of four repeated doses (2.5, 5, 25, and 50 mL/kg BW) of CU were tested and compared with three doses (10, 25, and 50 mg/kg BW) cyclophosphamide as a positive control and deionized water as the negative control. The results proved that cytological examination of CU was cytotoxic since a decrease in mitotic activity (16.8-1.1) was observed, since the significant reduction in cell proliferation in A. cepa L. and also in mice bone marrow cells. On the other hand, CU did not induce a clastogenic effect since no significant stickiness, fragment, multinucleoli were observed compared to the control group. Additionally, the data showed that CU decreased the CA when mice had received cyclophosphamide (25 mg BW) followed by CU doses. CU was found to be cytotoxic but no clastogenic effect. Furthermore, it possesses anticlastogenic properties. The observed results suggest that CU in whole or the metabolites present in CU could be a potent drug target. Further research is warranted to study the complete metabolites profiling and to study the molecular mechanisms.


Diversity, ecology and evolution of Archaea.
Compared to bacteria, our knowledge of archaeal biology is limited. Historically, microbiologists have mostly relied on culturing and single-gene diversity surveys to understand Archaea in nature. However, only six of the 27 currently proposed archaeal phyla have cultured representatives. Advances in genomic sequencing and computational approaches are revolutionizing our understanding of Archaea. The recovery of genomes belonging to uncultured groups from the environment has resulted in the description of several new phyla, many of which are globally distributed and are among the predominant organisms on the planet. In this Review, we discuss how these genomes, together with long-term enrichment studies and elegant in situ measurements, are providing insights into the metabolic capabilities of the Archaea. We also debate how such studies reveal how important Archaea are in mediating an array of ecological processes, including global carbon and nutrient cycles, and how this increase in archaeal diversity has expanded our view of the tree of life and early archaeal evolution, and has provided new insights into the origin of eukaryotes.


[Brain imaging in autism spectrum disorders. A review].
In the past two decades, an increasing number of functional and structural brain imaging studies has provided insights into the neurobiological basis of autism spectrum disorders (ASD). This article summarizes pertinent functional brain imaging studies addressing the neuronal underpinnings of ASD symptomatology (impairments in social interaction and communication, repetitive and restrictive behavior) and associated neuropsychological deficits (theory of mind, executive functions, central coherence), complemented by relevant structural imaging findings. The results of these studies show that although cognitive functions in ASD are generally mediated by the same brain regions as in typically developed individuals, the degree and especially the patterns of brain activation often differ. Therefore, a hypothesis of aberrant network connectivity has increasingly been favored over one of focal brain dysfunction.


The Role of CTGF in Inflammatory Responses Induced by Silica Particles in Human Bronchial Epithelial Cells.
Prolonged exposure to crystalline silica leads to persistent pulmonary inflammation and progressive fibrosis. Connective tissue growth factor (CTGF) has emerged as a potent proinflammatory and profibrotic regulator to participate in a variety of chronic inflammatory diseases. However, the role of CTGF in silica-induced pulmonary inflammation remains poorly understood. To explore the effect of CTGF on inflammatory responses caused by silica particles, human bronchial epithelial cells (16HBE) were transfected with CTGF siRNA and exposed to silica particles at concentrations of 0, 12.5, 25, 50, 100 μg/ml for 48 h. Intracellular CTGF mRNA and protein expressions were determined by RT-PCR and Western blotting, respectively. The levels of inflammatory cytokines including IL-8, TNF-α, IL-6, IL-1β, IL-17A and TGF-β1 were measured by ELISA kits. Silica particles induce significantly elevated intracellular CTGF mRNA expression in 16HBE cells in a dose-dependent manner when compared with blank control group (P < 0.05). The secretions of IL-8, TNF-α, IL-6 and IL-17A were also significantly increased by silica particles (P < 0.05). After exposure to 25 or 50 μg/ml silica particles, the expression of intracellular CTGF mRNA was significantly inhibited in 16HBE cells when transfected with CTGF siRNA (P < 0.05). The secreted levels of IL-8, TNF-α, IL-6 and IL-17A induced by silica particles were also significantly lower from CTGF siRNA-transfected cells than that from normal 16HBE cells (P < 0.05). Inhibition of CTGF gene attenuates silica-induced inflammatory responses in bronchial epithelial cells, suggesting that CTGF could be a pivotal regulator in the development of silica-induced inflammation.


Transcatheter arterial embolization in abdominal blunt trauma with active mesenteric bleeding: case series and review of literature.
To evaluate the safety and effectiveness of transcatheter arterial embolization (TAE) in treating traumatic mesenteric injuries with active bleeding, to report the outcome in a case series, and to compare the results with the existing data. All consecutive patients with active mesenteric bleeding due to blunt abdominal trauma referred to a level-one Trauma Center and treated by TAE were included; the related demographic and medical data were retrospectively reported. A literature review was conducted; all reported cases were collected and analysed together with our case series. A univariate analysis of risk factors for TAE failure, bowel necrosis, complication and length of stay was performed. Four consecutive patients were included. Technical success was 100%. One patient developed colon ischemia after the procedure and underwent surgical treatment; another presented transient mild renal failure and late respiratory failure. No 30-day mortality was reported. These results are consistent with those reported in literature. The analysis of our cases together with case collected from literature resulted in a case series of 25 patients. Univariate analysis showed colon as site of bleeding as a significant risk factor for bowel necrosis and older age as a significant risk factor for longer length of stay. TAE failure was not significantly associated neither with a higher complication rate nor with a higher length of stay. TAE is a safe and effective procedure to control mesenteric bleeding, thus should be considered, in selected cases and in appropriate setting, as an alternative to emergency surgery.


Mesothelioma and analysis of tissue fiber content.
The strong relationship between mesothelioma and asbestos exposure is well established. The analysis of lung asbestos burden by light and electron microscopy assisted to understand the increased incidence of mesothelioma in asbestos mining and consuming nations.The data on the occupational exposure to asbestos are important information for the purpose of compensation of occupational disease No. 4105 (asbestos-associated mesothelioma) in Germany.However, in many cases the patients have forgotten conditions of asbestos exposure or had no knowledge about the used materials with components of asbestos. Mineral fiber analysis can provide valuable information for the research of asbestos-associated diseases and for the assessment of exposure. Because of the variability of asbestos exposure and long latency periods, the analysis of asbestos lung content is a relevant method for identification of asbestos-associated diseases. Also, sources of secondary exposure, so called "bystander exposition" or environmental exposure can be examined by mineral fiber analysis.Household contacts to asbestos are known for ten patients (1987-2009) in the German mesothelioma register; these patients lived together with family members working in the asbestos manufacturing industry.Analysis of lung tissue for asbestos burden offers information on the past exposure. The predominant fiber-type identified by electron microscopy in patients with mesothelioma is amphibole asbestos (crocidolite or amosite). Latency times (mean 42.5 years) and mean age at the time of diagnose in patients with mesothelioma are increasing (65.5 years). The decrease of median asbestos burden of the lung in mesothelioma patients results in disease manifestation at a higher age.Lung dust analyses are a relevant method for the determination of causation in mesothelioma. Analysis of asbestos burden of the lung and of fiber type provides insights into the pathogenesis of malignant mesothelioma. The most important causal factor for the development of mesothelioma is still asbestos exposure.


Influence of cellulose nanocrystals (CNC) on permeation through intestinal monolayer and mucus model in vitro.
Cellulose nanocrystals (CNC) as a novel ingredient in foods and pharmaceuticals still lacks the safety and functionality information. We aimed to assess the absorption of CNC in small intestine and the effect on cell viability. In the second part, the impact of CNC on substance permeation through mucus layer, including the potential functionality in improving high blood cholesterol, was tested. No noticeable amount of CNC was found to penetrate through differentiated Caco-2 monolayer and in vitro mucus layer, and CNC had low toxicity on Caco-2 cell viability up to 10 mg/mL. CNC at 2 % (w/w) may affect the permeability of the mucus layer and larger molecules are more easily influenced. CNC may also alleviate hypercholesteremia by increasing viscosity of digesta, adsorbing cholesterol, and decreasing bile acids permeation. The results suggest CNC may not penetrate the small intestinal lining and may be used as a functional supplement.


Audio and video recording system for routine documentation of fluoroscopic procedures.
Because aviation and other reliable systems routinely record and analyze performance, the authors investigated the feasibility and utility of installing a system capable of recording image-guided procedures. An audio/video recording system adapted from those used in simulation laboratories was installed in a new pediatric interventional radiology suite. In this report, the authors describe the recording system as well as how it can be used to study radiation use during individual procedures. These results illustrate how routinely recording more than 300 procedures during the past 14 months and the detailed analysis of selected recordings can provide insights that lead to continual improvements in performance.


Quantitative structure-activity relationship (QSAR) for a series of novel cannabinoid derivatives using descriptors derived from semi-empirical quantum-chemical calculations.
Recent work implicating the cannabinoid receptors in a wide range of human pathologies has intensified the need for reliable QSAR models for drug discovery and lead optimization. Predicting the ligand selectivity of the cannabinoid CB(1) and CB(2) receptors in the absence of generally accepted models for their structures requires a ligand-based approach, which makes such studies ideally suited for quantum-chemical treatments. We present a QSAR model for ligand-receptor interactions based on quantum-chemical descriptors (an eQSAR) obtained from PM3 semi-empirical calculations for a series of phenyl-substituted cannabinoids based on a ligand with known in vivo activity against glioma [Duntsch, C.; Divi, M. K.; Jones, T.; Zhou, Q.; Krishnamurthy, M.; Boehm, P.; Wood, G.; Sills, A.; Moore. B. M., II. J. Neuro-Oncol., 2006, 77, 143] and a set of structurally similar adamantyl-substituted cannabinoids. A good model for CB(2) inhibition (R(2)=0.78) has been developed requiring only four explanatory variables derived from semi-empirical results. The role of the ligand dipole moment is discussed and we propose that the CB(2) binding pocket likely possesses a significant electric field. Describing the affinities with respect to the CB(1) receptor was not possible with the current set of ligands and descriptors, although the attempt highlighted some important points regarding the development of QSAR models.


Influence of siblings on adaptive behavior trajectories in autism spectrum disorder.
Siblings have the potential to shape the developmental trajectories of individuals with autism spectrum disorder. Having siblings has been associated with better social communication skills, less severe non-verbal communication deficits, and improved theory of mind abilities in autism spectrum disorder. This study examined the influence of the presence of a sibling, participant's position in the birth order, gender of the closest-age sibling, and gender match of the sibling dyad on the rate of growth in adaptive skills from ages 9 to 26 years among individuals with autism spectrum disorder and individuals with a history of non-spectrum developmental disorders. While all participants showed consistent gains in adaptive skill development as a function of age, there was variability in rates of growth based on sibling constellation group membership. Participants with at least one sibling, regardless of other sibling constellation factors, experienced greater rates of growth in adaptive behavior trajectories from childhood to adulthood than participants with no siblings across both Black and White participants, although differences were magnified in Black participants. Furthermore, among participants with siblings, those with male closest-age siblings and those with gender-matched siblings, irrespective of birth order position, demonstrated the steepest growth patterns in adaptive functioning within their respective groups over the 17-year period. Results suggest that siblings may serve an important role in improving the adaptive functioning trajectories and overall outcomes of individuals with autism spectrum disorder.


Changes in a motor pattern following cerebellar and olivary lesions in the squirrel monkey.
Squirrel monkeys were trained to perform ballistically initiated flexion movements of the forearm. It is shown that the motor output to agonist and antagonist subsequent to the onset of the movement depends continuously on the movement's parameters, primarily the velocity, as in man. The contribution of cerebellar activities to maintain this relationship was investigated by means of discrete lesions of the cerebellum and inferior olive. After lesions of the cerebellar nuclei, EMG activity of agonist and antagonist is no longer precisely structured. This result implies that activity of the cerebellar nuclei is necessary to maintain the gain and timing of the relationship between sensory inputs and motor outputs. Participation of the inferior olive is also required, since lesions of this structure mimic the effect of destruction of the cerebellar nuclei. Lesions of the cerebellar cortex, instead, affect mainly the timing of the motor output.


Right upper quadrant cytoreductive procedures and cardiophrenic lymph node resection in primary debulkig surgery for ovarian cancer.
Upfront debulking surgery followed by adjuvant chemotherapy still remains as the mainstay approach to patients with advanced ovarian cancer (Eisenhauer et al., 2006). Upper abdominal surgery is often required to achieve complete gross resection and there are several studies in the literature reporting increased survival, as well as a minimal but acceptable increase in morbidity, as a result of this shift in the surgical paradigm (Chi et al., 2009). Cardiophrenic lymph nodes (CPLNs), also referred to as paracardiac and supradiaphragmatic lymph nodes, are located just above the diaphragm. In diseases such as advanced ovarian cancer, where there is often considerable abdominal and peritoneal tumor burden, these lymph nodes can harbor metastases. These nodes are usually larger than 5 mm in diameter and are easily identified on computed tomography. Evidence suggests that this finding should not preclude primary debulking surgery (if resectable disease in the abdomen) as it is associated with an increased median overall survival even in stage IV disease (Cowan et al., 2017, Prader et al., 2016). We present a video highlighting one of the most commonly performed debulking procedures in the upper abdomen - right diaphragmatic peritoneal stripping (the patient had multiple small implants in both the diaphragmatic peritoneum and Morisońs Pouch peritoneum, so en bloc resection was performed) - followed by a transabdominal excision of an enlarged right cardiophrenic lymph node. The defect was closed with a 2-0 polypropylene running suture.


Rational design of medium supplementation strategy for improved influenza viruses production based on analyzing nutritional requirements of MDCK Cells.
Influenza vaccine production using cell culture technology has become popular nowadays. However, to meet the ever increasing demand of influenza vaccine, it is prerequisite to improve the yield of influenza virus in cells. To achieve this, in the present study, the nutritional requirements of MDCK cells in the virus production process were analyzed and a nutrient-feeding strategy was developed accordingly. Based on the consumption rates and corresponding concentration optimization, glucose and fast metabolized amino acids were supplemented into the maintaining medium at the time of infection. Compared with the non-supplemented culture, the average cell specific death rate during 0-48 h post-infection was 0.013 h(-1), which was 40.91% lower in the nutrient-supplemented culture. Total virus titer, HA antigen protein concentration and cell-specific virus yield were (1.88±0.23)×10(3) HA units/50μL, 11.70±0.22 μg/mL and (10.06±1.16)×10(3) virions/cell, respectively, which were 84.04±22.50%, 31.46±2.87% and 86.64±25.81% higher than those in the control, respectively. These data showed that the appropriate supplementation of nutrients during virus production process could reduce cell death, and improve cell-specific virus yield and total influenza virus output. This study laid foundation for the development of cell culture technology for influenza vaccine production.


Relationship between Sloan-Kettering virus expression and mouse follicular development.
Sloan-Kettering virus gene product (Ski) is an unique nuclear pro-oncoprotein and belongs to the ski/sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski/SnoN are important transcription regulators of the transforming growth factor-β (TGF-β) superfamily and function mainly through heterodimers. Since TGF-β superfamily are key regulators of follicle development and it has been previously shown that SnoN is also vital to follicle development, this research was conducted to clarify the relationship between Ski expression and mouse follicular development, in ovaries of neonatal and gonadotropin-induced immature mice by immunohistochemical and real-time PCR techniques. In postnatal mice, positive staining for Ski was highly detected in oocyte nuclei at postnatal day 1. With follicular development, the localization moved gradually from oocyte nuclei to perinuclear space and the total levels decreased. During the estrous cycle, Ski expression was apparent at proestrus and estrus, faint at metestrus, highest at diestrus. After injection of gonadotropin, Ski was found in perinuclear space and weak in oocyte nuclei. Following the initiation of luteinization, the expression of Ski was found in corpus luteum. Real-time PCR results also showed that Ski mRNA expression was opposite to ovulation-related genes during the cumulus expansion, with the development of the follicles, its expression level decreased. Ski is expressed in a specific manner during follicle development, ovulation and luteinization. So Ski might play essential roles in these processes especially during early follicular development.


Pulsed Field Ablation Using a Lattice Electrode for Focal Energy Delivery: Biophysical Characterization, Lesion Durability, and Safety Evaluation.
Pulsed field ablation (PFA) is a nonthermal energy that may provide safety advantages over radiofrequency ablation (RFA). One-shot PFA catheters have been developed for pulmonary vein isolation, but they do not permit flexible lesion sets. This study investigated a novel lattice-tip catheter designed for focal RFA or PFA ablation. The effects of PFA (biphasic, 24 amperes) were investigated in 25 swine using a lattice-tip catheter and system (Affera Inc). Step 1 (n=14) examined the feasibility to create atrial line of block and described its acute effects on the phrenic nerve and esophagus. Step 2 (n=7) examined the subacute effects of PFA on block durability, phrenic nerve, and esophagus ≥2 weeks. Step 3 compared the effects of PFA and RFA on the esophagus using a mechanical deviation model approximating the esophagus to the right atrium (n=4) and by direct ablation within its lumen (n=4). The effects of endocardial PFA and RFA on the phrenic nerve were also compared (n=10). Histological analysis was performed. PFA produced acute block in 100% of lines, achieved with 2.1 (1.3-3.2) applications/cm line. Histological analysis following (35 [18-37]) days showed 100% transmurality (thickness range 0.4-3.4 mm) with a lesion width of 19.4 (10.9-27.4 mm). PFA selectively affected cardiomyocytes but spared blood vessels and nervous tissue. PFA applied from the posterior atria (23 [21-25] applications) to the approximated esophagus (6 [4.5-14] mm) produced transmural lesions without esophageal injury. PFA (16.5 [15-18] applications) applied inside the esophageal lumen produced mild edema compared with RFA (13 [12-14] applications) which produced epithelial ulcerations. PFA resulted in no or transient stunning of the phrenic nerve (<5 minutes) without histological changes while RFA produced paralysis. PFA using a lattice-tip ablation catheter for focal ablation produced durable atrial lesions and showed lower vulnerability to esophageal or phrenic nerve damage compared with RFA.


Autologous blood as a submucosal fluid cushion for endoscopic mucosal therapies: results of an ex vivo study.
Submucosal injection of fluid is used to elevate lesions in order to prevent perforation, which is the most calamitous complication during endoscopic resection therapies. There are several injection options when performing mucosal elevation (normal saline (NS), sodium hyaluronate (SH), etc.). Submucosal injection of fresh, autologous blood offers some advantages because of its specific properties: corpuscular components ensure prolonged elevation and procoagulatory constituents prevent post-interventional bleeding. The purpose of this study was to compare the ex vivo performance of autologous blood as a submucosal fluid cushion (SFC) with that of NS, SH and DW (dextrose water). The proximal third of a resected porcine stomach was cut into squares. One millilitre NS, DW, SH and fresh porcine blood was injected into the submucosa. The height and duration of the submucosal injections were objectively measured during 1 h. Mucosal elevations were resected using an electro snare. The initial height and width of the mucosal elevations were comparable for SH and blood, and significantly higher compared with NS and DW. Mucosal elevation after injecting autologous blood persisted significantly longer compared with NS (p <0.05), but did not differ from hyaluronate. Histopathological examination of the resected specimen confirmed the appropriate submucosal injection of these substances. Submucosal injection of autologous blood with a standard endoscopic injection needle is possible and generates adequate mucosal elevation for the resection of high-quality specimens. This procedure could offer a "gratis" option for SFC as opposed to the expensive SH. Further clinical studies are needed to substantiate its use.


Effects of humic acids in vitro.
Humic acids are known for their overall positive health and productivity effects in animal feeding trials and, controversially, as an aetiological factor of cancer. We tried to assess the in vitro effect of humic acids from a selected source in Slovakia when used at recommended prophylactic dosage. We investigated antioxidant properties, enzymatic and non-enzymatic antioxidant defence system in liver mitochondria and cultured cancer cell lines in vitro. We observed a significant decrease in superoxide dismutase activity after humic acids treatment irrespective of dissolving in dimethyl sulphoxide or direct addition to mitochondria suspension in a respiration medium. Activities of other antioxidant enzymes measured, such as glutathione peroxidase and glutathione reductase, showed no significant differences from the control as well as the reduced glutathione content. Percentage of inhibition by humic acids of superoxide radical indicated lower efficacy compared with that of hydroxyl radical. Survival of six different cancer cells lines indicated that only the acute T lymphoblastic leukaemia cell line was sensitive to the tested humic acids. Despite relatively low solubility in aqueous solutions, humic acids from the selected source participated in redox regulation. By recapturing the radicals, humic acids reloaded the antioxidant defensive mechanism. Results from in vitro study conducted with humic acids from the natural source showed potential of these substances as promising immunity enhancing agents.


Preparation and evaluation of ascorbyl glucoside and ascorbic acid solid in oil nanodispersions for corneal epithelial wound healing.
The objective of this study was to develop and evaluate an effective topical formulation to promote corneal epithelial wound healing. Ascorbyl glucoside (AA-2G), a stable prodrug of AA, was formulated in solid in oil (S/O) nanodispersions by emulsifying AA-2G solutions in cyclohexane using Span 85 as an emulsifying agent and freeze-drying emulsions to produce AA-2G - surfactant complex. The complexes were then dispersed in castor oil to produce S/O nanodispersions which were evaluated in terms of their particle size, polydispersity index, encapsulation efficiency, morphology, physical stability as well as the transcorneal permeation and accumulation of AA-2G. The same preparation procedure was used to prepare S/O nanodispersions of AA. S/O nanodispersions of AA and AA-2G were formulated into oily drops that were tested for efficacy in promoting wound healing after corneal epithelial depredation. AA-2G was loaded efficiently in S/O nanodispersions (EE > 99%) in the form of spherical nanoparticles. S/O nanodispersions were physically stable and resulted in improved permeation (18x) and accumulation (7x) of AA-2G in transcorneal diffusion experiments in comparison to AA-2G solutions. Oily eye drops of AA-2G and AA showed no irritation and significant improvement in epithelial healing in vivo in comparison to AA-2G and AA solutions.


[Magnetic resonance imaging in perianal Crohn's disease].
Magnetic resonance imaging (MRI) was undertaken in a prospective study of 34 consecutive patients (21 women, 13 men; median age 31 [18-53] years) suspected of having active perianal Crohn's disease. The results of the investigation were compared with those obtained by independent observers on proctological and intraoperative examination (n = 31). A total of 58 fistulas and 21 abscesses were noted intraoperatively, 47 fistulas and all 21 abscesses by MRI, and 40 fistulas and 13 abscesses proctologically. The proctological examination proved to be more sensitive in demonstrating short subcutaneous or anovaginal fistulas (three of four subcutaneous and two of five anovaginal fistulas were not shown by MRI). Intersphincteric, ischiorectal and supralevator involvement was shown better by MRI. These results indicate that in perianal Crohn's disease MRI is a useful addition to proctological examination.


Scurvy: historically a plague of the sailor that remains a consideration in the modern intensive care unit.
We report the case of the case of a 56 year old female with sepsis on a background of rheumatoid arthritis and steroid use manifesting with overt clinical features of scurvy. Ascorbic acid assays were able to demonstrate severe deficiency and confirm a diagnosis of scurvy. Clinical resolution of signs and symptoms following commencement of vitamin C replacement was rapid. The intensivist and dietitian need to consider this diagnosis even in the first world setting, particularly in the presence of sepsis, inflammatory conditions, steroid use and importantly malnutrition.


Ultrasensitive detection of SARS-CoV-2 antigen using surface-enhanced Raman spectroscopy-based lateral flow immunosensor.
The fast spread and transmission of the coronavirus 2019 (COVID-19) has become one of serious global public health problems. Herein, a surface enhanced Raman spectroscopy-based lateral flow immunoassay (LFA) was developed for the detection of SARS-CoV-2 antigen. Using uniquely designed core-shell nanoparticle with embedded Raman probe molecules as the indicator to reveal the concentration of target protein, excellent quantitative performance with a limit of detection (LOD) of 0.03 ng/mL and detection range of 10-1000 ng/mL can be achieved within 15 min. Besides, the detection of spiked virus protein in human saliva was also performed with a portable Raman spectrometer, proposing the feasibility of the method in practical applications. This easy-to-use, rapid and accurate method would provide a point-of-care testing way as the ideal alternative for current detection requirement of virus-related biomarkers.


Age at initiation and frequency of screening to detect type 2 diabetes: a cost-effectiveness analysis.
No clinical trials have assessed the effects or cost-effectiveness of sequential screening strategies to detect new cases of type 2 diabetes. We used a mathematical model to estimate the cost-effectiveness of several screening strategies. We used person-specific data from a representative sample of the US population to create a simulated population of 325,000 people aged 30 years without diabetes. We used the Archimedes model to compare eight simulated screening strategies for type 2 diabetes with a no-screening control strategy. Strategies differed in terms of age at initiation and frequency of screening. Once diagnosed, diabetes treatment was simulated in a standard manner. We calculated the effects of each strategy on the incidence of type 2 diabetes, myocardial infarction, stroke, and microvascular complications in addition to quality of life, costs, and cost per quality-adjusted life-year (QALY). Compared with no screening, all simulated screening strategies reduced the incidence of myocardial infarction (3-9 events prevented per 1000 people screened) and diabetes-related microvascular complications (3-9 events prevented per 1000 people), and increased the number of QALYs (93-194 undiscounted QALYs) added over 50 years. Most strategies prevented a significant number of simulated deaths (2-5 events per 1000 people). There was little or no effect of screening on incidence of stroke (0-1 event prevented per 1000 people). Five screening strategies had costs per QALY of about US$10,500 or less, whereas costs were much higher for screening started at 45 years of age and repeated every year ($15,509), screening started at 60 years of age and repeated every 3 years ($25,738), or a maximum screening strategy (screening started at 30 years of age and repeated every 6 months; $40,778). Several strategies differed substantially in the number of QALYs gained. Costs per QALY were sensitive to the disutility assigned to the state of having diabetes diagnosed with or without symptoms. In the US population, screening for type 2 diabetes is cost effective when started between the ages of 30 years and 45 years, with screening repeated every 3-5 years. Novo Nordisk, Bayer HealthCare, [corrected] and Pfizer.


Correction to: Transcriptional and metabolite analysis reveal a shift in direct and indirect defences in response to spider‑mite infestation in cucumber (Cucumis sativus).
In the above mentioned publication, part of Fig. 1b was distorted (48 h after TSSM Infestation). The original article has been corrected and the proper version of Fig. 1 is also published here.


Trypsinogen isoforms in the ferret pancreas.
The domestic ferret (Mustela putorius furo) recently emerged as a novel model for human pancreatic diseases. To investigate whether the ferret would be appropriate to study hereditary pancreatitis associated with increased trypsinogen autoactivation, we purified and cloned the trypsinogen isoforms from the ferret pancreas and studied their functional properties. We found two highly expressed isoforms, anionic and cationic trypsinogen. When compared to human cationic trypsinogen (PRSS1), ferret anionic trypsinogen autoactivated only in the presence of high calcium concentrations but not in millimolar calcium, which prevails in the secretory pathway. Ferret cationic trypsinogen was completely defective in autoactivation under all conditions tested. However, both isoforms were readily activated by enteropeptidase and cathepsin B. We conclude that ferret trypsinogens do not autoactivate as their human paralogs and cannot be used to model the effects of trypsinogen mutations associated with human hereditary pancreatitis. Intra-pancreatic trypsinogen activation by cathepsin B can occur in ferrets, which might trigger pancreatitis even in the absence of trypsinogen autoactivation.


Partnership working in Africa: Shane Godbolt providing the links to the tapestry.
This paper explores the work that Shane Godbolt did in Africa in strengthening health information networks. It focuses on th role she played in supporting the Association for Health Information and Libraries in Africa (AHILA) and working with the Information Training Outreach Centre for Africa (ITOCA) to provide capacity building linkages. Some examples of such linkages extending beyond Africa to the United Kingdom are explored. It then concludes with reflections from authors on their own experiences of working with Shane Godbolt.


Microelectrode clusters enable therapeutic deep brain stimulation without noticeable side-effects in a rodent model of Parkinson's disease.
Deep Brain Stimulation (DBS) is an established treatment for motor symptoms in Parkinson's disease (PD). However, side effects often limit the usefulness of the treatment. To mitigate this problem, we developed a novel cluster of ultrathin platinum-iridium microelectrodes (n = 16) embedded in a needle shaped gelatin vehicle. In an established rodent PD-model (6-OHDA unilateral lesion), the clusters were implanted in the subthalamic area for up to 8 weeks. In an open field setting, combinations of microelectrodes yielding therapeutic effects were identified using statistical methods. Immunofluorescence techniques were used for histological assessments of biocompatibility. In all rats tested (n = 5), we found subsets of 3-4 microelectrodes which, upon stimulation (160 Hz, 60 μs pulse width, 25-40 μA/microelectrode), prompted normal movements without noticeable side effects. Other microelectrode subsets often caused side effects such as rotation, dyskinesia and tremor. The threshold (per microelectrode) to elicit normal movements strongly depended on the number of activated microelectrodes in the selected subset. The histological analysis revealed viable neurons close to the electrode contacts, minor microglial and astrocytic reactions and no major changes in the vasculature, indicating high biocompatibility. By contrast to the continuous and relatively large stimulation fields produced by existing DBS electrodes, the developed microelectrode cluster enables a fine-tuned granular and individualized microstimulation. This granular type of stimulation pattern provided powerful and specific therapeutic effects, free of noticeable side effects, in a PD animal model.


Effect of cyclosporine on hepatic energy status and on fructose metabolism after portacaval shunt in dog as monitored by phosphorus-31 nuclear magnetic resonance spectroscopy in vivo.
The effect of cyclosporin A on the hepatic energy status and intracellular pH of the liver and its response to a fructose challenge has been investigated using in vivo phosphorus-31 nuclear magnetic resonance spectroscopy in dogs. Three experimental groups were studied: (a) control dogs (n = 5), (b) dogs 4 days after the creation of an end-to-side portacaval shunt (n = 5), and (c) dogs 4 days after portacaval shunt and continuous infusion of cyclosporin A (4 mg/kg/day) by way of the left portal vein (portacaval shunt plus cyclosporin A, n = 5). The phosphorus-31 nuclear magnetic resonance spectra were obtained at 81 MHz using a Bruker BIOSPEC II 4.7-tesla nuclear magnetic resonance system equipped with a 40-cm horizontal bore superconducting solenoid. The phosphomonoesters (p less than 0.01), inorganic phosphate and ATP levels (p less than 0.05) were decreased significantly in portacaval shunt-treated and in portacaval shunt-plus-cyclosporin A-treated dogs compared with unshunted control dogs. After a fructose challenge (750 mg/kg body wt, intravenously), fructose-1-phosphate metabolism was reduced in portacaval shunt-treated dogs compared with either the normal or portacaval shunt-plus-cyclosporin A-treated dogs (p less than 0.05). Both portacaval shunt- and portacaval shunt-plus-cyclosporin A-treated dogs demonstrated a reduced decline in ATP levels after fructose infusion when compared with the controls (p less than 0.05). Immediately after the fructose challenge, the intracellular pH decreased from 7.30 +/- 0.03 to 7.00 +/- 0.05 in all animals (p less than 0.01) and then gradually returned to normal over 60 min.(ABSTRACT TRUNCATED AT 250 WORDS)


Total Knee Arthroplasty Complicated by Posterior Dislocation of a Polyethylene Insert and Neurovascular Compromise.
Dislocation of the polyethylene insert is a rare complication of total knee arthroplasty (TKA), which has only been described in a handful of case reports. Here, we describe a report of a 55-year-old woman presenting 13 months after the primary TKA with signs of neurovascular compromise. A magnetic resonance image showed posterior extrusion of her polyethylene insert causing a mass effect on the gastrocnemius muscle and the popliteal neurovascular bundle. A multidisciplinary team including a joint reconstruction surgeon, vascular surgeon, and nerve specialist performed a revision TKA with peroneal nerve decompression. The polyethylene insert was noted to be dislocated, rotated 90 degrees, and incarcerated in the posterior knee.


Aged Lumbar Extension Strength of Chronic Low Back Pain in Korean Population of 10-80 Years.
The purpose of this study was to find the basic data of medical and exercise therapy by indexing lumbar extension muscle strength of low back pain (LBP) patients. In this cross-sectional study, 3078 chronic LBP participants from The J hospital, Seoul, Republic of Korea, from 2003 to 2010 were enrolled. Maximum muscle strength was measured at maximum flexion angle and maximum extension angle according to range of motion (ROM) results. For each isometric test, participants were seated and secured in the MEDX (medx lumbar extension machine, Ocala, FL, USA) machine. The relative ROM (P=0.012) differed significantly among the aged groups in all participants. In addition, mean of strength (P<0.001), maximal of strength (P<0.001), mean of strength %BW (P<0.001) and maximal of strength %BW (P<0.001) are significant differences in all participants. The results of multiple regression analysis was the 'model A', maximal of strength for 32.1% of the variance in weigh, body mass index and range of motion. In addition, 'model B' was 30.4%, 'model C' was 28.8%, 'model D' was 28.5%, 'model E' was 21.7%, and 'model F' was 23.5% of the variance in weigh, body mass index and range of motion. We found the three predictor (weight, BMI, and ROM) variables accounted for 32.1% of the variance in maximal of strength %BW, the highest in < 29 yr groups. Our data indicate the basic data of medical and exercise therapy by indexing lumbar extension muscle strength of LBP patients.


Global inflammation predicts cardiovascular risk in women: a report from the Women's Ischemia Syndrome Evaluation (WISE) study.
Measurement of C-reactive protein (CRP), a marker of inflammation, is recommended to improve cardiovascular disease (CVD) risk stratification. However, no studies have collectively evaluated how inflammatory markers cluster empirically and relate to angiographic coronary artery disease and CVD events. From the WISE study, 580 women with fasting plasma samples of inflammatory markers (interleukin [IL]-6, IL-18, tumor necrosis factor alpha, transforming growth factor beta, CRP, serum amyloid A [SAA], and intercellular adhesion molecules) were analyzed over a median of 4.7 years follow-up. All women were referred for coronary angiography (1996-2000) to evaluate suspected myocardial ischemia. In factor analysis, a "proinflammation" factor (cluster) loaded on IL-6, CRP, and SAA (r = 0.63-0.87); a "proinflammation and anti-inflammation" cluster loaded on IL-18 and tumor necrosis factor alpha (r = 0.72, 0.77); and an "immunosuppressive" factor loaded singly on transforming growth factor beta (r = 0.96). No cluster was independently associated with angiographic coronary artery disease. However, quartile increases of the "proinflammation" cluster (IL-6, CRP, and SAA) yielded death rates of 2.6%, 7.2%, 13.1%, 26.6%, respectively (P < .0001). Women with > or = 2 of 3 proinflammation markers in the upper quartile had an adjusted relative risk of death of 4.21 (95% CI 1.91-9.25), a higher conferred risk than any single marker alone, all of which were roughly equally predictive. Although IL-6, CRP, and SAA all predict CVD risk in women, development of global measures of inflammation and simply counting the number of markers with high levels improve CVD risk stratification. In addition, results indicate that the adverse impact of inflammation may be largely through other mechanisms than promotion of atherogenesis (ie, destabilization of vulnerable plaques).


Study on the SFRP4 gene polymorphism and expression in prostate cancer.
Prostate cancer is a heterogeneous disease and considered to be the most commonly diagnosed cancer. SFRP4 gene acts as Wnt antagonist in the Wnt signalling pathway, thereby playing an important role in carcinogenesis. The aim of the present study was to investigate two single-nucleotide polymorphisms: c.958 C>A (rs1802073) and c.1019 G>A (rs1802074) in the SFRP4 gene and its expression in prostate cancer. A sample size of 100 cases and 100 age-matched controls were recruited for the study. Statistical analysis revealed the heterozygous GA genotype of rs1802074 significantly increased in cases when compared to controls. Analysis of sFRP4 expression based on the genotypes showed a significantly increased expression for the heterozygous GA and homozygous AA genotypes in cases when compared to the controls. Fold change was calculated using 2-ΔΔCT method and the results showed that there were a 3.4 and 4.5 fold increase in the sFRP4 expression for GA and AA genotypes, respectively. Our results suggest that the rs1802074 polymorphism in SFRP4 gene may be associated with the risk of prostate cancer.


[Antiedema activity of a non-steroid physiological anti-inflammatory agent (author's transl)].
The antiedema activity of experimental Lysoartrosi, a biological preparation, was demonstrated against edemas induced by carrageenin, formalin, dextran, serotonin, egg albumin and kaolin. Lysoartrosi, containing a mixture of polypeptides, is thus, the only preparation which is active against such a wide spectrum of inflammatory agents.


Establishment of a prognostic signature based on fatty acid metabolism genes in HCC associated with hepatitis B.
Hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) is one of the most common and deadly cancer and often accompanied by varying degrees of liver damage, leading to the dysfunction of fatty acid metabolism (FAM). This study aimed to investigate the relationship between FAM and HBV-associated HCC and identify FAM biomarkers for predicting the prognosis of HBV-associated HCC. Gene Set Enrichment Analysis (GSEA) was used to analyze the difference of FAM pathway between paired tumor and adjacent normal tissue samples in 58 HBV-associated HCC patients from the Gene Expression Omnibus (GEO) database. Next, 117 HBV-associated HCC patients from The Cancer Genome Atlas (TCGA) database were analyzed to establish a prognostic signature based on 42 FAM genes. Then, the prognostic signature was validated in an external cohort consisting of 30 HBV-associated HCC patients. Finally, immune infiltration analysis was performed to evaluate the FAM-related immune cells in HBV-associated HCC. As a result, FAM pathway was clearly downregulated in tumor tissue of HBV-associated HCC, and survival analysis demonstrated that 12 FAM genes were associated with the prognosis of HBV-associated HCC. Lasso-penalized Cox regression analysis identified and established a five-gene signature (ACADVL, ACAT1, ACSL3, ADH4 and ECI1), which showed effective discrimination and prediction for the prognosis of HBV-associated HCC both in the TCGA cohort and the validation cohort. Immune infiltration analysis showed that the high-risk group, identified by FAM signature, of HBV-associated HCC had a higher ratio of Tregs, which was associated with the prognosis. Collectively, these findings suggest that there is a strong connection between FAM and HBV-associated HCC, indicating a potential therapeutic strategy targeting FAM to block the accumulation of Tregs into the tumor microenvironment of HBV-associated HCC.


Computerized Intervention in Primary Care for Women Veterans with Sexual Assault Histories and Psychosocial Health Risks: a Randomized Clinical Trial.
Sexual assault is a public health concern for women and is associated with subsequent psychosocial health risks of posttraumatic stress disorder (PTSD), hazardous drinking, and intimate partner violence (IPV). Sexual assault is associated with social stigma and other barriers shown to inhibit one from seeking mental health care. Digital health technologies may overcome these barriers. To test the impact of a brief computerized intervention delivered in primary care to reduce health risks and increase mental health treatment utilization among women with histories of sexual assault and current health risks. The Safe and Healthy Experiences (SHE) program was tested in a randomized controlled trial with N = 153 women veterans at a Veterans Health Administration (VHA) medical center, and they completed assessments at baseline, 2 months, and 4 months. SHE is a brief motivational interviewing and psychoeducation-based computerized intervention. SHE was compared to a screen and referral-only control condition. Health risks were measured via validated self-report instruments. Treatment initiation and utilization were measured via self-report and chart review. SHE did not impact women's number of health risks (all p's > .05). However, women randomized to SHE showed significantly greater increases in treatment use compared to women in the control group, as measured by chart review (χ2 (1, n = 153) = 4.38, p = .036, rs = .16), and self-report (χ2 (1, n = 130) = 5.89, p = .015, rs = .21). SHE was found to be an acceptable intervention. SHE was effective in improving mental health treatment initiation and utilization compared to a control group. Computer-based interventions to address sexual trauma and its consequences are acceptable, are highly scalable, and can add value to primary care with little cost or increase in provider time. Clinicaltrials.gov identifier NCT02957747.


A novel sunshine duration-based photothermal time model interprets the photosensitivity of flower maturity of pecan cultivars.
Although it is well-known and established that light plays important roles in plant development, up to now, there is no substantial improvements in how to deal with the light factor of spring phenology under natural condition. By monitoring the local meteorologic data and mature dates of two types (male and female) of flower from four pecan cultivars during 9 years, it was found that the complementary pattern of growing degree day and sunshine duration helped to maintain a threshold of driving force related to the maturity of pecan flower during 9 years. A novel photothermal time model based on the linear combination of growing degree day and sunshine duration was then proposed and validated to interpret the variance of mature dates of pecan cultivars. Comparative analysis showed that the new model had made extremely significant improvements to the traditional thermal time model. In addition, this model introduced the conversion coefficient K, which quantified the effect of light on the flowering drive, and revealed the differences of base temperature among cultivars. This was the first time that sunshine duration instead of photoperiod was adopted to develop into a verified model on spring phenological event of tree species. It will help to model the spring phenologies of other tree species more reasonably.


Auditory testing outcomes with hearing aids in patients with auditory neuropathy spectrum disorder.
The objective of this study is to evaluate the audiologic outcomes with hearing aids in pediatric patients with auditory neuropathy spectrum disorder (ANSD) using the Infant Toddler-Meaningful Auditory Integration Scale (IT-MAIS), and the Ling 6 Sound Test (Ling 6). Case series. Single tertiary care academic medical center. All pediatric patients with a confirmed diagnosis of ANSD on Auditory Brainstem Response (ABR) testing who presented to a single tertiary medical center between September 2008 and September 2018 were included. Only patients that underwent Infant Toddler-Meaningful Auditory Integration Scale (IT-MAIS) and/or Ling 6 Sound Test (Ling 6) were included in the study. Audiologic testing performed after cochlear implantation was excluded. 60 pediatric patients with ANSD were analyzed. There were 10 patients included in the study with documented hearing aid use who underwent IT-MAIS and/or Ling 6 testing. Average IT-MAIS score improved by 20.4% after initial or extended trial of amplification. Similarly, average Ling 6 score improved from 3.6 to 4.8 after initial or extended trial of amplification. The four patients who did not receive amplification had higher average IT-MAIS and Ling 6 scores. In most children with ANSD, IT-MAIS and Ling 6 Sound Test scores improved with initial hearing aid use and over time with extended hearing aid use. Long-term prospective, multi-institutional studies are needed to determine the impact of the natural history of ANSD, comorbidities, and socioeconomic variables on auditory function testing results in children with ANSD using hearing aids.


Hearing preservation following surgical removal of meningiomas affecting the temporal bone.
It is a common clinical impression that preservation of hearing is more often achieved when removing a meningioma than a similarly sized acoustic tumor. However, relatively few reports have focused on postoperative hearing results after meningioma removal, and detailed audiometric data are not commonly provided, particularly in the neurosurgical literature. During the past 16 years, 56 meningiomas affecting the temporal bone have been surgically removed at the House Ear Clinic. Hearing preservation was attempted in 16 (29%) of the 56 cases, and these were the focus of this study. The primary presenting symptom was otologic in 67% of these cases, including hearing loss as the primary symptom in 27%. Measurable postoperative hearing was present in 11 (92%) of 12 patients with postoperative audiograms available, and 8 (67%) of 12 patients had good hearing postoperatively. Hearing was preserved near the preoperative level (within 10 dB speech reception threshold and 15% speech discrimination) in 75% of cases.


Downregulation of microRNA-146a in diabetes, obesity and hypertension may contribute to severe COVID-19.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is able to produce an excessive host immune reaction and may leads to severe disease- a life-threatening condition occurring more often in patients suffering from comorbidities such as hypertension, diabetes and obesity. Infection by human corona viruses highly depends on host microRNA (miR) involved in regulation of host innate immune response and inflammation-modulatory miR-146a is among the first miRs induced by immune reaction to a virus. Moreover, recent analysis showed that miR-146 is predicted to target at the SARS-CoV-2 genome. As the dominant regulator of Toll-like receptors (TLRs) downstream signaling, miR-146a may limit excessive inflammatory response to virus. Downregulation of circulating miR-146a was found in diabetes, obesity and hypertension and it is reflected by enhanced inflammation and fibrosis, systemic effects accompanying severe COVID-19. Thus it could be hypothesized that miR-146a deficiency may contribute to severe COVID-19 state observed in diabetes, obesity and hypertension but further investigations are needed.


Improved ZAP-70 assay using two clones, multiple methods of analysis and clinical correlation.
In a companion methodological study, we compared two anti-ZAP-70 clones (1E7.2 AF 488 and SBZAP PE) and four selected methods of analysis. Clinical correlations are required for validation. Multicolor flow-cytometric evaluation of ZAP-70, CD38, CD69, CD26, CD49d, and CD27 was tested in 45 untreated-CLL patients. Four methods of ZAP-70 expression analysis and a scoring system were designed. A correlation analysis between ZAP-70 score, immunoglobulin heavy chain variable (IGHV) mutational status, fluorescence in situ hybridization, and these biomarkers was undertaken. There is a strong correlation between ZAP-70 expression and IGHV mutational status. The scoring system for a single reagent (P = 0.0006 or 0.0002) favors the use of multiple methods of analysis. The combined score was substantially equivalent (P = 0.0003). There was also a correlation with del 13q14 (P = 0.017) and trisomy12 (P = 0.011). A correlation for CD38 and ZAP-70 score was seen using both 1E7.2 AF488 and SBZAP PE when ≥20% or ≥7% cutoff was used. A positive correlation was seen for CD49d expression using both reagents. CD26 showed a correlation with ZAP-70 expression, but it was dependent upon the method of analysis. CD69 and CD27 showed no statistically significant correlation. In our study population, ZAP-70 expression is the better predictor of the IGHV mutational status. The correlation analysis confirms that the use of four methods of analysis with a single reagent or both reagents is superior to the use of a single method of analysis. The routine use of CD38, CD49d, and CD26 will require standardization.


FTIR and UV spectroscopy studies of triplex formation between pyrimidine methoxyethylphosphoramidates alpha-oligodeoxynucleotides and ds DNA targets.
The ability of non-ionic methoxyethylphosphoramidate (PNHME) alpha-oligodeoxynucleotides (ODNs), alpha dT(15) and alpha dCT dodecamer, to form triplexes with their double-stranded DNA targets was evaluated. Thermal stability of the formed complexes was studied by UV thermal denaturation and the data showed that these PNHME alpha-ODNs formed much more stable triplexes than phosphodiester (PO) beta-ODNs did (Delta Tm = + 20 degrees C for alpha dCT PNHME). In addition, FTIR spectroscopy was used to determine the base pairing and the strand orientations of the triplexes formed by alpha dT(15) PNHME compared to phosphodiester ODNs with beta or alpha anomeric configuration. While beta dT(15) PO failed to form a triplex with a long beta dA(n) x beta dT(n) duplex, the Tm of the Hoogsteen part of the triplex formed by alpha dT(15) PNHME reached 40 degrees C. Moreover alpha dT(15) PNHME displaced the beta dT(15) strand of a shorter beta dA(15) x beta dT(15) duplex. The alpha dCT PNHME and alpha dT(15) PNHME third strands were found antiparallel in contrast to alpha dT(15) PO which is parallel to the purine strand of their duplex target. The uniform preferential Hoogsteen pairing of the nucleotides alpha dT and alpha dC combining both replacements might contribute to the improve stability of the triplexes.


Ultrastructure of mouse vallate taste buds: II. Cell types and cell lineage.
The lifespan of cells in the mouse taste bud was examined with high-voltage electron microscopic (HVEM) autoradiography (ARG) after giving a single injection of 3H-thymidine. Animals were killed at 1 hour, 6 hours, 12 hours, 24 hours, and then daily up through 10 days postinjection. Lingual tissues were prepared for HVEM ARG so that we could identify and characterize labeled cells. Four categories of taste cells were identified: basal, dark, intermediate, and light cells. Basal cells were polygonal cells located near the basolateral sides of the taste buds and were characterized primarily by the presence of filaments attached to the nuclear envelope. Dark and light cells had the typical features described by previous authors. Intermediate cells had features in between those of dark and light cells. Over 90% of the cells labeled in the first 2 days following injection of 3H-thymidine were basal cells. Labeled dark cells appeared 6 hours after injection, reached their peak incidence at the fourth day postinjection, and then gradually decreased. Labeled intermediate cells were identified after the appearance of dark cells (12 hours) and reached a peak incidence at the fifth day after injection of 3H-thymidine. Lastly, labeled light cells were first observed on the fourth day postinjection and continued to increase until the tenth day, when they constituted 45% of the labeled cells. These data support the hypothesis that there is one cell line in the mouse vallate taste bud that undergoes morphological changes in its lifespan.


An econometric analysis of SARS and Avian Flu on international tourist arrivals to Asia.
This paper compares the impacts of SARS and human deaths arising from Avian Flu on international tourist arrivals to Asia. The effects of SARS and human deaths from Avian Flu are compared directly according to the number of human deaths. The nature of the short run and long run relationship is examined empirically by estimating a static line fixed effect model and a difference transformation dynamic model, respectively. Empirical results from the static fixed effect and difference transformation dynamic models are consistent, and indicate that both the short run and long run SARS effect have a more significant impact on international tourist arrivals than does Avian Flu. In addition, the effects of deaths arising from both SARS and Avian Flu suggest that SARS is more important to international tourist arrivals than is Avian Flu. Thus, while Avian Flu is here to stay, its effect is currently not as significant as that of SARS.


Human and rat adrenal glycerol kinase: subcellular distribution and bisubstrate kinetics.
The subcellular distribution of adrenal glycerol kinase in man and rat are reported and the bisubstrate kinetics of the soluble enzyme are compared in these two species. The specific activity of glycerol kinase in human whole adrenal homogenate (145 microU/mg protein) was 3 times that found in rat whole adrenal homogenate (48 microU/mg protein). In both species 8% of the total glycerol kinase activity was associated with the nuclear pellet fraction. In human, 62% of the total activity was soluble, while 24% was associated with the postnuclear particulate fraction. Rat glycerol kinase activity was also predominantly soluble: 69% of the total activity was soluble and 13% was in the postnuclear particulate fraction. The apparent Km for glycerol in soluble adrenal glycerol kinase was similar in both species, 2.8 microM in human and 3.1 microM in rat. The apparent Km for ATP in soluble human adrenal glycerol kinase was 22.0 microM. In rat the enzyme did not appear to follow Michaelis-Menten kinetics with ATP as substrate. The Vmax for the soluble enzyme was similar in both human and rat. This report provides a background to biochemical investigations on human glycerol kinase deficiency, an inborn error of metabolism which may be characterized by adrenal hypoplasia and insufficiency.


Preferential Infection of α4β7+ Memory CD4+ T Cells During Early Acute Human Immunodeficiency Virus Type 1 Infection.
Establishment of persistent human immunodeficiency virus type 1 (HIV-1) reservoirs occurs early in infection, and biomarkers of infected CD4+ T cells during acute infection are poorly defined. CD4+ T cells expressing the gut homing integrin complex α4β7 are associated with HIV-1 acquisition, and are rapidly depleted from the periphery and gastrointestinal mucosa during acute HIV-1 infection. Integrated HIV-1 DNA was quantified in peripheral blood mononuclear cells obtained from acutely (Fiebig I-III) and chronically infected individuals by sorting memory CD4+ T-cell subsets lacking or expressing high levels of integrin β7 (β7negative and β7high, respectively). HIV-1 DNA was also assessed after 8 months of combination antiretroviral therapy (cART) initiated in Fiebig II/III individuals. Activation marker and chemokine receptor expression was determined for β7-defined subsets at acute infection and in uninfected controls. In Fiebig I, memory CD4+ T cells harboring integrated HIV-1 DNA were rare in both β7high and β7negative subsets, with no significant difference in HIV-1 DNA copies. In Fiebig stages II/III and in chronically infected individuals, β7high cells were enriched in integrated and total HIV-1 DNA compared to β7negative cells. During suppressive cART, integrated HIV-1 DNA copies decreased in both β7negative and β7high subsets, which did not differ in DNA copies. In Fiebig II/III, integrated HIV-1 DNA in β7high cells was correlated with their activation. β7high memory CD4+ T cells are preferential targets during early HIV-1 infection, which may be due to the increased activation of these cells.


Toxic effects of HgCl2 on activities of SOD, AchE and relative expression of SOD, AChE, CYP1A1 of zebrafish.
In order to explore the impact of mercury on fish, zebrafish were exposed in various levels of HgCl2 and the superoxide dismutase (SOD), acetylcholinesterase (AChE) activities were measured after 1 and 7 days exposure. Muscle RNA was also extracted for semi-quantitative RT-PCR determination of the expression of AChE, CYP1A1, Cu/Zn-SOD. It indicated that 96 h-LC50 of HgCl2 to zebrafish is 244.06 µg L(-1). The AChE activity was decreased on 1st day and 7th day. The SOD activity was almost unchanged on 1st day, it showed a dose-response effect on 7th day. The expression of AChE was down-regulated on 1st day, it is consistent with the AChE activity, but the expression was showed a significant difference between exposed group and the control group on 7th day. The expression of CYP1A1 was decreased compared with the control group on 1st day. After 7 day exposure, the expression of CYP1A1 was up-regulated in 0.1LC(50) group, and down-regulated in 0.4LC(50) group, 0.8LC(50) group, showing a stimulation effect at low concentration and inhibition at high concentration. The expression of Cu/Zn-SOD was down-regulated on 1st day, also showing an effect of stimulation at low concentration and inhibition at high concentrations. Thus HgCl2 was highly toxic to zebrafish, both the enzymatic activities and the gene transcription were changed significantly. But the change of gene transcription is earlier than enzymes activities. It indicates that gene transcript can be used as an early warning for environmental risk assessment.


Nocturnal melatonin profiles before and one year after beginning shift-work.
Nocturnal serum melatonin profiles were determined twice for seven single women, during their time of employment as nurses (baseline), and after one year (follow up), in order to investigate the effects of shift-work on nocturnal melatonin secretion. All subjects were working in the same hospital under an irregularly rotating three-shift system. Five (5) mL blood samples were drawn six times at 2 h intervals between 20:00-06:00 hours under dim light conditions (< 50 lux). The same sampling procedures were repeated the following year. The results showed pronounced inter-individual differences in melatonin concentrations. There was a trend towards increasing maximum melatonin concentration (MAX melatonin) at follow up, with a similar tendency seen in summed melatonin (the sum of six measured melatonin concentrations). A trend was also seen towards increasing melatonin ratio at 06:00 hours (the percentage of melatonin concentration at 06:00 hours by summed melatonin) at follow up. Melatonin concentration at 06:00 hours was significantly higher at follow up, and a significant correlation between Morningness-Eveningness score (M-E score) at baseline and increased summed melatonin at follow up was also seen. These results suggest that: nocturnal melatonin secretion does not significantly increase after beginning shift-work; and that greater increases in melatonin secretion at follow up are found in subjects with higher M-E scones (increased morning type). With more subjects, however, there may be significant increase in MAX melatonin and/or summed melatonin in the follow-up study.


Motor denervation of tumors of the head and neck: changes in MR appearance.
We reviewed the MR appearance of motor denervation of the third (mandibular) division of the trigeminal nerve (V3) and of the hypoglossal nerve. Six cases of tumor of the head and neck with motor denervation were retrospectively evaluated. These comprised two patients with V3 denervation, three patients with hypoglossal nerve denervation, and one patient with both V3 and hypoglossal denervation. The observation was conducted for 6 to 44 months after onset. In denervated muscles, changes in signal intensity in T(1)- and T(2)-weighted images, degree of contrast enhancement, and volume of muscle were estimated during the follow-up period. In all cases of V3 denervation, the muscles showed no change in signal intensity in the T(1)-weighted images up to three months after onset. In two cases of hypoglossal denervation, the tongues appeared ipsilaterally hyperintense in the T(1)-weighted images within the first three months. In one case with V3 denervation and two cases with hypoglossal denervation, denervated muscles appeared hyperintense in the T(2)-weighted images up to three months after onset. At three months after denervation, the signal intensities of all motor-denervated areas increased in both T(1)- and T(2)-weighted images. Postcontrast T(1)-weighted images obtained within the first three months displayed contrast enhancement of all denervated muscles. In three cases of V3 denervation, the volumes of the affected muscles were reduced after the first three months. In three cases of hypoglossal denervation, the ipsilateral volume of the tongue decreased at three months after onset. Up to three months after onset, the denervated muscles appeared hyperintense in the T(2)-weighted images and contrast enhancement in postcontrast T(1)-weighted images before fatty infiltration and volume loss were apparent. Familiarity with the MR appearance of denervated muscles accompanying tumors of the head and neck is important to avoid confusion with inflammatory or neoplastic processes.


A novel and secure technique of stemmed acetabular cup implantation in complex hip reconstructions: a comparative study and technical note.
The stemmed acetabular cup (SAC) can be considered in several complicated situations. This type of prosthesis uses the iliac isthmus, which is most often preserved, thus allowing solid anchorage. Its implantation is prone to mechanical complications intraoperatively, such as extra-isthmic effraction, fracture, and early aseptic loosening. The aim of our study was to compare a new technique using a flexible motorized reamer combined with isthmus catheterization (Group A) versus the standard technique deploying a rigid reamer alone (Group B). A retrospective analysis was carried out of the SACs implanted at our hospital. The main evaluation criterion was visualization of extra-isthmic effraction (EIE) of the stem. Fifty-six SACs (Integra Lepine®, Genay, France) were implanted, 31 in group A and 25 in group B. The mean follow-up was 21.6 months ± 22.3. This study found fewer EIEs using a flexible motorized reamer (0 versus 5, p = 0.014), less perioperative bleeding (p = 0.054), and less aseptic loosening in group A (p = 0.034). There was no difference in terms of infection or other mechanical complications post-operatively between the two groups. Using iliac catheterization combined with a flexible motorized reamer when implanting a SAC helps to avoid EIE and it also appears to reduce complications such as peri-operative bleeding and early aseptic loosening. We recommend use of this surgical technique rather than a rigid reamer alone, and this new technique should allow widespread and more accessible use of this specific type of implant.


Beta-actin is a target for transglutaminase activity at synaptic endings in chicken telencephalic cell cultures.
Transglutaminases are Ca(2+)-dependent enzymes that catalyse the covalent cross-linking of protein-bound glutamine and lysine residues, which can stabilise proteins or protein aggregates. In the brain, elevated expression levels and activity of transglutaminases are known to be linked with several neurodegenerative diseases. However, little is known about the physiological functions of transglutaminases in the central nervous system. In this study, we examined the expression and activity of transglutaminase 1 in chicken telencephalic cell cultures. We observed a cytosolic expression of transglutaminase 1 in telencephalic neurons. However, transglutaminase 1 activity was restricted to synaptic endings. Transglutaminase targets in the cultured cells were characterised via a biotinylation assay and β-actin was identified as a substrate. Furthermore, we were able to show that β-actin is a target for the activity of recombinant human transglutaminase 1 in vitro. We propose a mechanism where neuronal transglutaminase 1 is activated by synaptic activity-dependent influx of calcium ions and thereupon catalyse the formation of an intramolecular cross-link in β-actin, thereby stabilising the actin cytoskeleton against depolymerising effects. In this way, transglutaminase 1 could modulate actin-dependent plasticity events at synaptic endings.


M6A Classification Combined With Tumor Microenvironment Immune Characteristics Analysis of Bladder Cancer.
Studies have shown that N6-methyl adenosine (m6A) plays an important role in cancer progression; however, the underlying mechanism of m6A modification in tumor microenvironment (TME) cell infiltration of bladder cancer remains unclear. This study aimed to investigate the role of m6A modification in TME cell infiltration of bladder cancer. The RNA expression profile and clinical data of bladder cancer were obtained from The Cancer Genome Atlas and Gene Expression Omnibus. We assessed the m6A modification patterns of 664 bladder cancer samples based on 20 m6A regulators through unsupervised clustering analysis and systematically linked m6A modification patterns to TME cell infiltration characteristics. Gene ontology and gene set variation analyses were conducted to analyze the underlying mechanism based on the assessment of m6A methylation regulators. Principal component analysis was used to construct the m6A score to quantify m6A modification patterns of bladder cancer. The genetic and expression alterations in m6A regulators were highly heterogeneous between normal and bladder tissues. Three m6A modification patterns were identified. The cell infiltration characteristics were highly consistent with the three immune phenotypes, including immune rejection, immune inflammation, and immune desert. The biological functions of three m6A modification patterns were different. Cox regression analyses revealed that the m6A score was an independent signature with patient prognosis (HR = 1.198, 95% CI: 1.031-1.390). Patients with a low-m6A score were characterized by increased tumor mutation burden, PD-L1 expression, and poorer survival. Patients in the low-m6A score group also showed significant immune responses and clinical benefits in the CTLA-4 immunotherapy cohort (p =0.0069). The m6A methylation modification was related to the formation of TME heterogeneity and complexity. Assessing the m6A modification pattern of individual bladder cancer will improve the understanding of TME infiltration characteristics.


Predicting multiple conformations via sequence clustering and AlphaFold2.
AlphaFold2 (AF2) 1 has revolutionized structural biology by accurately predicting single structures of proteins. However, a protein's biological function often depends on multiple conformational substates2, and disease-causing point mutations often cause population changes within these substates3,4. We demonstrate that clustering a multiple sequence alignment (MSA) by sequence similarity enables AF2 to sample alternate states of known metamorphic proteins with high confidence. Using this method, AF-Cluster, we investigated the evolutionary distribution of predicted structures for the metamorphic protein KaiB5, and found that predictions of both conformations were distributed in clusters across the KaiB-family. We used nuclear magnetic resonance (NMR) spectroscopy to confirm a surprising AF-Cluster prediction: a cyanobacteria KaiB variant is stabilized in the opposite state than the more widely-studied variant. To test AF-Cluster's sensitivity to point mutations, we designed and experimentally verified a set of 3 mutations predicted to flip KaiB from Rhodobacter sphaeroides from the ground to the fold-switched state. Finally, screening for alternate states in protein families without known fold-switching identified a putative alternate state for the oxidoreductase Mpt53 in M. tuberculosis. Further development of such bioinformatic methods in tandem with experiments will likely have profound impact on predicting protein energy landscapes, essential for illuminating biological function.


Genetic-code evolution for protein synthesis with non-natural amino acids.
The genetic encoding of synthetic or "non-natural" amino acids promises to diversify the functions and structures of proteins. We applied rapid codon-reassignment for creating Escherichia coli strains unable to terminate translation at the UAG "stop" triplet, but efficiently decoding it as various tyrosine and lysine derivatives. This complete change in the UAG meaning enabled protein synthesis with these non-natural molecules at multiple defined sites, in addition to the 20 canonical amino acids. UAG was also redefined in the E. coli BL21 strain, suitable for the large-scale production of recombinant proteins, and its cell extract served the cell-free synthesis of an epigenetic protein, histone H4, fully acetylated at four specific lysine sites.


Inflammation versus host defense in obesity.
Obesity is characterized by a state of low-grade, chronic inflammation. Wang et al. (2014) report that immune cells from obese mice have decreased production of IL-22, a cytokine involved in immune responses and inflammation, and reveal therapeutic effects of exogenous IL-22 against obesity-linked metabolic dysfunctions.


Comparison of strategies for the construction of libraries of artificial antibodies.
Construction of libraries of artificial antibodies has been reported by several groups of investigators. Various forms of antibody fused to surface protein, cpII, are expressed on the surface of filamentous phage. Since phages that encode desired antibodies can be easily grown in Escherichia coli after selection with target antigens, the phage-display antibody system appears to be very useful for various biological purposes. In this brief review, recent progress in research into the production of artificial antibodies is summarized and the strategies used for the construction of libraries of artificial antibodies are compared.


Healing in the new millennium: bone stimulators: an overview of where we've been and where we may be heading.
Electromagnetic fields and their uses in bone healing have been fairly well studied, with most results showing improvement in healing of both bone and cartilage. Most supportive data are found in relation to the spine, femur, and tibia, but there is increasing evidence for its use in the foot and ankle for treatment of nonunions and as an adjunctive device in arthrodeses, particularly in high-risk populations. There are varying data and a significant variety of quality in the current research and publications concerning the use of electrical bone stimulation in the treatment of the foot and ankle. Thus, there is a definite need for further investigation and high-quality study designs to determine the most effective treatment modalities and pathologies best used with bone stimulation. Bone stimulation should be viewed as an adjunctive procedure in which the surgeon optimizes the high-risk patient both medically or surgically whenever possible. But when used appropriately, bone stimulation has the potential to influence outcomes and aid in bone healing when complications arise and in high-risk populations.


Sodium cromoglycate (intal) in the treatment of vernal keratoconjunctivitis and allergic conjunctivitis.
Intal drops (sodium cromoglycate) relieved symptoms of vernal conjunctivitis in all the 19 patients we studied, reducing the need for steroids although the response varied. Intal was also effective in all 11 patients with acute or subacute seasonal allergic conjunctivitis, but in only 4 of 11 patients with mild chronic conjunctivitis associated with atopy. No side-effects were noted other than mild irritation in some cases.


Racial and demographic disparities in emergency department utilization for mental health concerns before and during the COVID-19 pandemic.
This study investigated whether emergency department (ED) visits for mental health concerns increased during the COVID-19 pandemic, taking a health disparities lens. ED encounters from the only academic medical center in Mississippi were extracted from March-December 2019 and 2020, totaling 2,842 pediatric (ages 4-17) and 17,887 adult (ages 18-89) patients. Visits were coded based on primary ED diagnosis. For adults, there were fewer depression/anxiety ED visits during the pandemic, not moderated by any demographic factor, but no differences for serious mental illness or alcohol/substance use. For youth, there were significantly fewer ED visits for behavior problems during the pandemic among children in the lower socioeconomic status (SES) category; there were no differences for depression/anxiety. Regardless of year, adults in the lower SES category were more likely to visit the ED for mental health, Black adults were less likely to visit the ED for depression/anxiety or alcohol/substance use, and Black children were less likely to visit the ED for behavioral concerns. Results suggest that access to outpatient and telehealth services remains critical for mental health care during the pandemic and underline the importance of race- and SES-related factors in use of the ED for mental health concerns beyond the pandemic.


Target-promoted specific activation of m6A-DNAzyme for SPEXPAR-amplified and highly sensitive non-label electrochemical assay of FTO demethylase.
The demethylase of fat mass and obesity related protein (FTO) is critical to regulate the dynamic N6-methyladenosine (m6A) modification of eukaryotic mRNAs, and its overexpression has found to be closely related to the initiation of several cancers. On the basis of a target-promoted specific activation of DNAzyme strategy coupled with self-primer exponential amplification reaction (SPEXPAR) cycles and DNA supersandwich assemblies, the highly sensitive and label-free electrochemical FTO assay approach is established. The modification of the catalytic core nucleobase of the DNAzyme probe by m6A can inhibit its cleavage activity. The presence of target FTO catalyzes the elimination of the methyl group to restore the DNAzyme activity, which cleaves the hairpin substrates to trigger the SPEXPAR for yielding many ssDNAs. The capture of these DNAs on the sensor electrode leads to the initiation of supersandwich assembly formation of long dsDNAs. Tremendous electrochemical signal probe of [Ru(NH3)6]Cl3 are then absorbed on these dsDNAs to produce highly amplified catalytic currents with the assistance of K3[Fe(CN)6] for detecting trace FTO with 63.1 fM detection limit. Furthermore, the sensor can be employed for selective assay of FTO in cell lysates, revealing the great potential of this sensing strategy for biomedical and biological study applications.


Structural, electronic, optical and thermodynamic properties of AlAuO2 and AlAu094Fe006O2 compounds scrutinized by density functional theory (DFT).
In this study, density functional theory (DFT) simulations have been used to study the structural, electrical and optical properties of AlAuO2 and AlAu094Fe006O2. Initially, the estimated bandgap of AlAuO2 0.45, 0.486, 0.419 and 2.49 eV in Perdew-Burke-Ernzerhof (PBE), Revised Perdew-Burke-Ernzerhof (RPBE), PBE for solids (PBE sol) and Becke three-parameter Lee-Yang-Parr (B3LYP) method respectively while AlAu094Fe006O2 has a zero-band gap after 6 % Fe doping. Then, density of state (DOS) and partial density of state (PDOS) were studied to determine the characteristics of the various orbitals of AlAuO2. The bonding characteristics and thermal stability of this crystal are determined by the Mulliken population charge and thermos physical parameters. Band edge of AlAuO2 was calculated which revealed that the AlAuO2 has suitable oxidation and reduction potential to degrade the contamination. A remarkable absorption has recorded for both AlAuO2 and AlAu094Fe006O2 in visible and ultraviolet region and capability to utilize photocatalytic dye degradation and hydrogen production through water splitting.