Sex-dependent effect of a low neurosteroid environment and intrauterine growth restriction on foetal guinea pig brain development.
Progesterone and its neuroactive metabolite, allopregnanolone, are present in high concentrations during pregnancy, but drop significantly following birth. Allopregnanolone influences foetal arousal and enhances cognitive and behavioural recovery following traumatic brain injury. Inhibition of allopregnanolone synthesis increases cell death in foetal animal brains with experimental hypoxia. We hypothesised that complications during pregnancy, such as early or preterm loss of placental steroids and intrauterine growth restriction (IUGR), would disrupt the foetal neurosteroid system, contributing to poor neurodevelopmental outcomes. This study aimed to investigate the effects of chronic inhibition of allopregnanolone synthesis before term and IUGR on developmental processes in the foetal brain. Guinea pig foetuses were experimentally growth restricted at mid-gestation and treated with finasteride, an inhibitor of allopregnanolone synthesis. Finasteride treatment reduced foetal brain allopregnanolone concentrations by up to 75% and was associated with a reduction in myelin basic protein (MBP) (P = 0.001) and an increase in glial fibrillary acidic protein expression in the subcortical white matter brain region (P < 0.001). IUGR resulted in decreased MBP expression (P < 0.01) and was associated with a reduction in the expression of steroidogenic enzyme 5α-reductase (5αR) type 2 in the foetal brain (P = 0.061). Brain levels of 5αR1 were higher in male foetuses (P = 0.008). Both IUGR and reduced foetal brain concentrations of allopregnanolone were associated with altered expression of myelination and glial cell markers within the developing foetal brain. The potential role of neurosteroids in protecting and regulating neurodevelopmental processes in the foetal brain may provide new directions for treatment of neurodevelopmental disorders in infants who are exposed to perinatal insults and pathologies.


[Genetic characteristics and physical organization of the R-plasmid pBS52 with a broad range of bacterial hosts].
The genetic and physical data on Pseudomonas aeruginosa plasmid pBS52 coding for the resistances to ampicillin, streptomycin and sulfonamids have been obtained. This conjugative plasmid is transferable to a broad range of gram-negative bacterial hosts and compatible with the broad host-range plasmids from all known incompatibility groups. The plasmid size has been determined (38 Kb) and a physical map has been constructed using restriction endonucleases EcoRI, EcoRV, BamHI, BglII, PstI, PvuII, SalI, SlaI. The presence of a fragment, approximately 200 bp in size, which contains the sites for many of widely used restriction endonucleases is a characteristic feature of the plasmid pBS52.


Minimal T-cell-stimulatory sequences and spectrum of HLA restriction of immunodominant CD4+ T-cell epitopes within hepatitis C virus NS3 and NS4 proteins.
The hepatitis C virus (HCV)-specific CD4+ T-cell response against nonstructural proteins is strongly associated with successful viral clearance during acute hepatitis C. To further develop these observations into peptide-based vaccines and clinical immunomonitoring tools like HLA class II tetramers, a detailed characterization of immunodominant CD4+ T-cell epitopes is required. We studied peripheral blood mononuclear cells from 20 patients with acute hepatitis C using 83 overlapping 20-mer peptides covering the NS3 helicase and NS4. Eight peptides were recognized by > or = 40% of patients, and specific CD4+ T-cell clones were obtained for seven of these and three additional, subdominant epitopes. Mapping of minimal stimulatory sequences defined epitopes of 8 to 13 amino acids in length, but optimal T-cell stimulation was observed with 10- to 15-mers. While some epitopes were presented by different HLA molecules, others were presented by only a single HLA class II molecule, which has implications for patient selection in clinical trials of peptide-based immunotherapies. In conclusion, using two different approaches we identified and characterized a set of CD4+ T-cell epitopes in the HCV NS3-NS4 region which are immunodominant in patients achieving transient or persistent viral control. This information allows the construction of a valuable panel of HCV-specific HLA class II tetramers for further study of CD4+ T-cell responses in chronic hepatitis C. The finding of immunodominant epitopes with very constrained HLA restriction has implications for patient selection in clinical trials of peptide-based immunotherapies.


De novo design of anti-variant COVID-19 vaccine.
Recent studies highlight the effectiveness of hybrid Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) vaccines combining wild-type nucleocapsid and Spike proteins. We have further enhanced this strategy by incorporating delta and omicron variants' spike protein mutations. Both delta and omicron mark the shifts in viral transmissibility and severity in unvaccinated and vaccinated patients. So their mutations are highly crucial for future viral variants also. Omicron is particularly adept at immune evasion by mutating spike epitopes. The rapid adaptations of Omicron and sub-variants to spike-based vaccines and simultaneous transmissibility underline the urgency for new vaccines in the continuous battle against SARS-CoV-2. Therefore, we have added three persistent T-cell-stimulating nucleocapsid peptides similar to homologous sequences from seasonal Human Coronaviruses (HuCoV) and an envelope peptide that elicits a strong T-cell immune response. These peptides are clustered in the hybrid spike's cytoplasmic region with non-immunogenic linkers, enabling systematic arrangement. AlphaFold (Artificial intelligence-based model building) analysis suggests omitting the transmembrane domain enhances these cytoplasmic epitopes' folding efficiency which can ensure persistent immunity for CD4+ structural epitopes. Further molecular dynamics simulations validate the compact conformation of the modeled structures and a flexible C-terminus region. Overall, the structures show stability and less conformational fluctuation throughout the simulation. Also, the AlphaFold predicted structural epitopes maintained their folds during simulation to ensure the specificity of CD4+ T-cell response after vaccination. Our proposed approach may provide options for incorporating diverse anti-viral T-cell peptides, similar to HuCoV, into linker regions. This versatility can be promising to address outbreaks and challenges posed by various viruses for effective management in this era of innovative vaccines.


Treatment of Intracranial Infection Caused by Methicillin-Resistant Staphylococcus epidermidis with Linezolid Following Poor Outcome of Vancomycin Therapy: A Case Report and Literature Review.
The pharmacokinetic/pharmacodynamic (PK/PD) parameter for evaluating the efficacy of vancomycin is now recommended to target an AUC/MIC (area under the curve, AUC; minimum inhibitory concentration, MIC) ratio of 400 to 600, and trough concentration should not be used as a substitute. We report a case of intracranial infection caused by methicillin-resistant Staphylococcus epidermidis (MRSE), which was sensitive to vancomycin (MIC=2µg/mL) and linezolid (MIC=4µg/mL). The trough concentration of vancomycin in serum was 18.3 µg/mL, and the vancomycin concentration in CSF was 5.0 µg/mL, all within normal range. However, the AUC/MIC ratio was calculated to be 125 mg·h·L-1, unable to reach target AUC/MIC. Vancomycin was replaced with linezolid after 36 days of treatment due to poor outcome, and the patient was eventually cured. Further, 23 cases of intracranial methicillin-resistant Staphylococcus aureus (MRSA) or methicillin-resistant coagulase-negative Staphylococcus (MRCoNS) infections were reported, of which 1 case with MRSA had a vancomycin MIC of 1 µg/mL, while the remaining 22 cases had vancomycin MICs >1 µg/mL. The linezolid-containing regimen was used after drug susceptibility results or if the initial treatment failed, leading to recovery in 19 patients, microbial clearance in 3 patients, and treatment failure in 1 case. In conclusion, vancomycin dosing should be based on AUC-guided dosing and monitoring. When the vancomycin MIC of MRSA/MRCoNS is >1 µg/mL, the target AUC/MIC may not be achieved. In such cases, linezolid can effectively be considered as a good alternative to vancomycin.


Use of medicinal plants by black women: ethnography study in a low-income community.
Objective To explore beliefs, values and practices related to the use of medicinal plants among low-income black families. Method The research method was ethnography and the participant observation process was done in a low-income community in the peripheral area of the City of São Paulo. Twenty black women were interviewed. Results Two cultural sub-themes, I do use medicines that I learned to make with my mother and with religious practitioners to care for diseases and Home medicines are to treat problems that are not serious, and the cultural theme I do use home medicines to treat simple diseases because I always have them at my disposal, they are free and I don't need a medical prescription represent beliefs, values, and practices related to the use of medicinal plants among low-income black families. Conclusion The development of such practices, which can hide ethnic and social vulnerability, reveals the resilience of low-income black women in the process of confronting problems during the health-illness process.


ER contact sites direct late endosome transport.
Endosomes shuttle select cargoes between cellular compartments and, in doing so, maintain intracellular homeostasis and enable interactions with the extracellular space. Directionality of endosomal transport critically impinges on cargo fate, as retrograde (microtubule minus-end directed) traffic delivers vesicle contents to the lysosome for proteolysis, while the opposing anterograde (plus-end directed) movement promotes recycling and secretion. Intriguingly, the endoplasmic reticulum (ER) is emerging as a key player in spatiotemporal control of late endosome and lysosome transport, through the establishment of physical contacts with these organelles. Earlier studies have described how minus-end-directed motor proteins become discharged from vesicles engaged at such contact sites. Now, Raiborg et al. implicate ER-mediated interactions, induced by protrudin, in loading plus-end-directed motor kinesin-1 onto endosomes, thereby stimulating their transport toward the cell's periphery. In this review, we recast the prevailing concepts on bidirectional late endosome transport and discuss the emerging paradigm of inter-compartmental regulation from the ER-endosome interface viewpoint.


[Esomeprasol and rabeprasol effects on esophageal acidification in patients with gastroesophageal reflux disease, intensively metabolizing inhibitors of proton pump].
To compare action of different esomeprasol and rabeprasol doses on esophageal acidification in patients with gastroesophageal reflux disease (GERD). A cross-over study was made in 27 GERD patients (12 males and 15 females). It included esophagoduodenoscopy with biopsy for detection of H-pylori and 24-h pH-metry of esophageal content. Rabeprasol and esomeprasol efficacy was compared for daily doses 20 mg of each (stage I) and rabeprasol 20 mg and esomeprasol 40 mg (stage II). All the patients proved H-pylori positive. Before the treatment mean daily time of marked esophageal acidification was 29-36%. At stage I significant differences between the drugs were absent. At stage If esomeprasol was more effective from the first day of therapy both in relation to time to effect and its intensity. Esomeprasol provided a fast and noticeable correction of esophageal acidification. This relieves symptoms of GERD and accelerates esophageal mucosa epithelization.


Stimulus Sensitive Smart Nanoplatforms: An Emerging Paradigm for the Treatment of Skin Diseases.
Over the past century, the prevalence of skin diseases has substantially increased. These diseases present a significant physical, emotional and socio-economic burden to the society. Such conditions are also associated with a multitude of psychological traumas to the suffering patients. The effective treatment strategy implicates targeting of drugs to the skin. The field of drug targeting has been revolutionized with the advent of nanotechnology. The emergence of stimulus-responsive nanoplatforms has provided remarkable control over fundamental polymer properties for external triggers. This enhanced control has empowered pioneering approaches in the treatment of chronic inflammatory skin diseases. Our aim was to investigate the studies on smart nanoplatforms that exploit the altered skin physiology under diseased conditions and provide site-specific controlled drug delivery. All literature search regarding the advances in stimulus sensitive smart nanoplatforms for skin diseases was done using Google Scholar and Pubmed. Various stimuli explored lately for such nano platforms are pH, temperature, light and magnet. Although, the scientists have actively taken up this research topic but there are still certain lacunaes associated which have been discussed in this review. Further, an interdisciplinary collaboration between the healthcare providers and pharmacists is a pivotal requirement for such systems to be available for patients.


Role of hepatic Annexin A6 in fatty acid-induced lipid droplet formation.
Annexin A6 (AnxA6) has been implicated in the regulation of endo-/exocytic pathways, cholesterol transport, and the formation of multifactorial signaling complexes in many different cell types. More recently, AnxA6 has also been linked to triglyceride storage in adipocytes. Here we investigated the potential role of AnxA6 in fatty acid (FA) - induced lipid droplet (LD) formation in hepatocytes. AnxA6 was associated with LD from rat liver and HuH7 hepatocytes. In oleic acid (OA) -loaded HuH7 cells, substantial amounts of AnxA6 bound to LD in a Ca2+-independent manner. Remarkably, stable or transient AnxA6 overexpression in HuH7 cells led to elevated LD numbers/size and neutral lipid staining under control conditions as well as after OA loading compared to controls. In contrast, overexpression of AnxA1, AnxA2 and AnxA8 did not impact on OA-induced lipid accumulation. On the other hand, incubation of AnxA6-depleted HuH7 cells or primary hepatocytes from AnxA6 KO-mice with OA led to reduced FA accumulation and LD numbers. Furthermore, morphological analysis of liver sections from A6-KO mice revealed significantly lower LD numbers compared to wildtype animals. Interestingly, pharmacological inhibition of cytoplasmic phospholipase A2α (cPLA2α)-dependent LD formation was ineffective in AnxA6-depleted HuH7 cells. We conclude that cPLA2α-dependent pathways contribute to the novel regulatory role of hepatic AnxA6 in LD formation.


Functional Enhancers As Master Regulators of Tissue-Specific Gene Regulation and Cancer Development.
Tissue-specific transcription is critical for normal development, and abnormalities causing undesirable gene expression may lead to diseases such as cancer. Such highly organized transcription is controlled by enhancers with specific DNA sequences recognized by transcription factors. Enhancers are associated with chromatin modifications that are distinct epigenetic features in a tissue-specific manner. Recently, super-enhancers comprising enhancer clusters co-occupied by lineage-specific factors have been identified in diverse cell types such as adipocytes, hair follicle stem cells, and mammary epithelial cells. In addition, noncoding RNAs, named eRNAs, are synthesized at super-enhancer regions before their target genes are transcribed. Many functional studies revealed that super-enhancers and eRNAs are essential for the regulation of tissue-specific gene expression. In this review, we summarize recent findings concerning enhancer function in tissue-specific gene regulation and cancer development.


Epitope-tagged yeast strains reveal promoter driven changes to 3'-end formation and convergent antisense-transcription from common 3' UTRs.
Epitope-tagging by homologous recombination is ubiquitously used to study gene expression, protein localization and function in yeast. This is generally thought to insulate the regulation of gene expression to that mediated by the promoter and coding regions because native 3' UTR are replaced. Here we show that the 3' UTRs, CYC1 and ADH1, contain cryptic promoters that generate abundant convergent antisense-transcription in Saccharomyces cerevisiae. Moreover we show that aberrant, truncating 3' -end formation is often associated with regulated transcription in TAP-tagged strains. Importantly, the steady-state level of both 3' -truncated and antisense transcription products is locus dependent. Using TAP and GFP-tagged strains we show that the transcriptional state of the gene-of-interest induces changes to 3' -end formation by alternative polyadenylation and antisense transcription from a universal 3' UTR. This means that these 3' UTRs contains plastic features that can be molded to reflect the regulatory architecture of the locus rather than bringing their own regulatory paradigm to the gene-fusions as would be expected. Our work holds a cautionary note for studies utilizing tagged strains for quantitative biology, but also provides a new model for the study of promoter driven rewiring of 3' -end formation and regulatory non-coding transcription.


Putting the pieces into place: Properties of intact zinc metallothionein 1A determined from interaction of its isolated domains with carbonic anhydrase.
Mammalian metallothioneins (MTs) bind up to seven Zn(2+) using a large number of cysteine residues relative to their small size and can act as zinc-chaperones. In metal-saturated Zn7-MTs, the seven zinc ions are co-ordinated tetrahedrally into two distinct clusters separated by a linker; the N-terminal β-domain [(Zn3Cys9)(3-)] and C-terminal α-domain [(Zn4Cys11)(3-)]. We report on the competitive zinc metalation of apo-carbonic anhydrase [CA; metal-free CA (apo-CA)] in the presence of apo-metallothionein 1A domain fragments to identify domain specific determinants of zinc binding and zinc donation in the intact two-domain Znn-βαMT1A (human metallothionein 1A isoform; n=0-7). The apo-CA is shown to compete effectively only with Zn2-3-βMT and Zn4-αMT. Detailed modelling of the ESI mass spectral data have revealed the zinc-binding affinities of each of the zinc-binding sites in the two isolated fragments. The three calculated equilibrium zinc affinities [log(KF)] of the isolated β-domain were: 12.2, 11.7 and 11.4 and the four isolated α-domain affinities were: 13.5, 13.2, 12.7 and 12.6. These data provide guidance in identification of the location of the strongest-bound and weakest-bound zinc in the intact two-domain Zn7βαMT. The β-domain has the weakest zinc-binding site and this is where zinc ions are donated from in the Zn7-βαMT. The α-domain with the highest affinity binds the first zinc, which we propose leads to an unscrambling of the cysteine ligands from the apo-peptide bundle. We propose that stabilization of the intact Zn6-MT and Zn7-MT, relative to that of the sum of the separated fragments, is due to the availability of additional cysteine ligand orientations (through interdomain interactions) to support the clustered structures.


An evaluation of poor LMA CTrach views with a fibreoptic laryngoscope and the effectiveness of corrective measures.
The LMA CTrach++ system has integrated fibreoptics to enable viewing of the glottis to aid tracheal intubation via a laryngeal mask conduit. Earlier evaluations of this system had shown high success rates of insertion, ventilation and tracheal intubation, but frequent difficulty or failure to view the glottis. We evaluated CTrach views in 100 patients. In 69 patients with partial or no views of the glottis, we evaluated the causes with the aid of an Olympus LF-DP fibreoptic laryngoscope passed through the CTrach. In those with poor views, we tested the effectiveness of simple corrective measures to improve the views and optimize CTrach placement. These included pushing the CTrach deeper in, partial withdrawal by 6 cm and reinsertion to correct epiglottic downfolding, withdrawal by 1 cm and applying forward lift to correct obstruction by the arytenoids, and complete removal to clean the fibreoptic ports followed by reinsertion when secretions were the problem. Downfolding of the epiglottis caused poor views in 57 patients, which was correctable in 51. Obstruction of the view by the arytenoids occurred in seven patients and by secretions in five, all of which were correctable. With the Ctrach, we were able to ventilate the lungs in all patients, view the glottis in 94 and intubate the trachea in 97 patients. Despite the ability to ventilate the lungs in all patients with the CTrach, poor initial views were very frequent, indicating suboptimal placement, although most poor views can be improved, leading to a high intubation success rate.


[Compound Chamomile and Lidocaine Hydrochloride Gel applied at different time-windows for premature ejaculation: A prospective single-center randomized controlled study].
To investigate the efficacy and safety of Compound Chamomile and Lidocaine Hydrochloride Gel (CCLH) (Kamistad) applied at different time-windows on premature ejaculation (PE). This prospective study included 72 PE patients treated by application of CCLH to the glans and penile body in our hospital from February to October 2021. According to the time of drug administration before insertion into the vagina, we randomly divided the patients into a 5-minute group (n = 39) and a 15-minute group (n = 33). Before and after 1 and 2 weeks of treatment, we compared the intravaginal ejaculation latency time (IELT), PE diagnostic tool (PEDT) score, quality of life, and adverse reactions between the two groups of patients. Totally 62 of the patients completed the follow-up, 35 in the 5-minute group and 27 in the 15-minute group, and all showed significant improvement in IELT (P < 0.01) and PEDT score (P < 0.05) after treatment compared with the baseline. No allergic reactions, such as redness and swelling, developed at the application site in any of the patients, and no adverse significant effect was observed on the erectile hardness in 61 of the cases. Six cases showed increased erectile hardness instead. Fifty-seven of the patients experienced no obvious penile numbness or reduced sexual satisfaction, and all could complete their sexual activities. Compound Chamomile and Lidocaine Hydrochloride Gel applied at different time-windows is effective on PE, with a 5-minute rapid onset of action before intercourse, and no obvious adverse effects.


Drug delivery to the brain: how can nanoencapsulated statins be used in the clinic?
Statins are used for the primary and secondary prevention of cardiovascular disease by inhibiting cholesterol synthesis in the liver. Statins have also noncholesterol-related effects, called pleiotropic effects, which arise from statins' anti-inflammatory, immunomodulatory and antioxidant properties. These effects are especially attractive for the treatment of various brain diseases ranging from stroke to neurodegenerative diseases. Still, low brain concentrations after oral drug administration hinder the clinical application of statins in these pathologies. Pharmaceutical nanotechnologies may offer a solution to this problem, as local or targeted delivery of nanoencapsulated statins may increase brain availability. This special report rapidly summarizes the potential of statins in the treatment of brain diseases and the pharmaceutical nanotechnologies that could provide a viable approach to enable these indications.


Renewable hybrid nanocatalyst from magnetite and cellulose for treatment of textile effluents.
A hybrid catalyst was prepared using cellulose nanofibrils and magnetite to degrade organic compounds. Cellulose nanofibrils were isolated by mechanical defibrillation producing a suspension used as a matrix for magnetite particles. The solution of nanofibrils and magnetite was dried and milled resulting in a catalyst with a 1:1 ratio of cellulose and magnetite that was chemically and physically characterized using light, scanning electron and transmission electron microscopies, specific surface area analysis, vibrating sample magnetometry, thermogravimetric analysis, Fourier transform infrared spectroscopy, X-ray diffraction, catalytic potential and degradation kinetics. Results showed good dispersion of the active phase, magnetite, in the mat of cellulosic nanofibrils. Leaching and re-use tests showed that catalytic activity was not lost over several cycles. The hybrid material produced was tested for degradation of methylene blue dye in Fenton-like reactions resulting in a potential catalyst for use in degradation of organic compounds.


Drug-eluting stents versus arterial myocardial revascularization in patients with diabetes mellitus.
The introduction of drug-eluting stents significantly reduced restenosis and reinterventions in patients undergoing percutaneous coronary interventions. This study compares results of Cypher stenting with those of surgical arterial revascularization in patients with diabetes mellitus. From May 2002 through May 2005, 523 consecutive diabetic patients underwent myocardial revascularization: 244 underwent percutaneous coronary interventions incorporating drug-eluting stents, and 279 were treated surgically. All single-vessel patients in the surgical group were treated with the left internal thoracic artery, and most multivessel patients were treated with 2 internal thoracic arteries. After propensity score matching, 2 groups (93 patients each) were used to compare the 2 revascularization modalities. The number of coronary vessels treated per patient was higher in the surgical group (2.72 vs 1.75, P < .001). Follow-up ranged between 6 and 42 months (mean, 19 months). Overall mortality (early and late) was 3.2% in the surgical group and 2.2% in the Cypher group (P = .65). Two-year angina-free survival and reintervention-free survival (Kaplan-Meier) of the surgical group were 88% and 95%, respectively, compared with 47.8% (P = .001) and 83.6% (P = .01), respectively, in the percutaneous coronary intervention group. Cox proportional hazards modeling revealed assignment to the Cypher group to be the only predictor of reintervention (odds ratio, 3.86; 95% confidence interval, 1.25-11.9). Assignment to the Cypher group (hazard ratio, 5.92; 95% confidence interval, 2.96-11.87) and insulin treatment (hazard ratio, 2.06; 95% confidence interval, 1.06-4.02) were independent predictors of angina recurrence. The midterm clinical outcome of diabetic patients who underwent surgical arterial revascularization is better than that of patients undergoing percutaneous coronary intervention treated with drug-eluting stents.


Overexpression of Notch3 is associated with metastasis and poor prognosis in osteosarcoma patients.
Notch signaling abnormalities are associated with the development of various tumors, including hematopoietic and epithelium-derived tumors. However, the role of Notch signaling in tumors originating from mesenchymal cells is unclear. The effect of Notch3 expression on the prognosis of osteosarcoma and its role and mechanism in osteosarcoma cells have never been reported. In this study, we performed a clinicopathological analysis of 70 cases of osteosarcoma, with primary focus on survival. Osteosarcoma cell lines MTH and U2OS were used. After knockdown of Notch3 by lentiviral transfection and siRNA, the cell cycle, cell viability, and wound healing capacity were assessed. Subsequently, the Transwell assay was performed, and the expression levels of hairy and enhancer of split-1 (Hes1) and matrix metalloproteinase 7 (MMP7) were detected by RT-PCR and Western blot assay. The expression of MMP7 was also detected after knockdown of Hes1. Animal experiments were performed by injecting the cell lines MTH of Notch3 knockdown into mice tail veins and comparing the development of lung metastasis with the control group. Comparison of survival curves showed that Notch3 expression significantly impacts patient survival. Additionally, multivariate analysis revealed that Notch3 is an independent prognostic factor for osteosarcoma. In in vivo experiments, osteosarcoma-associated pulmonary metastasis in nude mice was reduced after Notch3 silencing. The expression of downstream effector molecule, Hes1, and that of the invasion and metastasis-associated proteolytic enzyme, MMP7, were reduced, and MMP7 was further decreased by Hes1 knockdown in in vitro experiments. Notch3 is a prognostic factor for osteosarcoma and might regulate its invasion and metastasis through the downstream target gene Hes1 and effector MMP7.


Experimental evaluation of some controversial points regarding the A-V conduction disturbances in the clinical His bundle electrogram.
Although the recordings of the His potential have been shown to be very useful in the study of cardiac conduction and rhythm disturbances, there remain some problems regarding the clinical His bundle electrogram. Some of the studies were performed to solve in the perfused dog hearts and in situ dog hearts, and consequently the following results were obtained. In some experiments, recording of the His potential could not be achieved even in the perfused hearts in spite of the preserved A-V conduction, suggesting that, as experienced sometimes in the clinical cases, inability to record the His potential might not be due to the technical failure or the anatomical abnormalities of the His bundle. On the other hand, real "splitting of H" occurred in association with the increase in the duration and decrease in the amplitude of the His potential after the ligation of the anterior septal artery. In addition, the true Mobitz II type A-H block, and Wenckebach type intra-Hisian and H-V block could be produced experimentally.


Novel route for silylation of silica gel and aliphatic amines immobilization based on microwave-assisted solvent free synthesis and their applications for Cu(II) and Fe(III) removal from natural water samples.
This article describes a new route for silica gel silylation and immobilization of aliphatic amines based on microwave-assisted solvent free synthesis to produce new solid phase extractors. The mode of synthesis was optimized under microwave conditions and achieved in a short time without using solvents. The produced phases named: silica gel- monoamine (SG-MA), silica gel- ethylenediamine (SG-EDA) and silica gel- diethylenetriamine (SG-DETA). The selectivity of these phases towards the uptake of Cu(II) and Fe(III) was checked using batch equilibration technique. Microwave radiation power and time of radiation were optimized to obtain the highest metal uptake values. The novel synthesized silica amine phases were characterized using Fourier transform infrared spectra and scanning electron microscope. The effects of different parameters including, hydrogen ion concentration, initial metal ion concentration, mass of the phase and shaking time on binding capacities of both Cu(II) and Fe(III) were explored. Results of sorption isotherms of the phases were better fitted with the Langmuir model (r² ≥ 0.950). In addition, the kinetics data were best fitted with the pseudo-second-order type (r² = 0.999). Application of SG-MA for removal of Cu(II)- and Fe(III)-spiked natural water samples was achieved satisfactorily using batch experiments. The results were found to refer to superior recovery percentages (90.0-97.01 ± 0.010-0.521%) with no significant matrix interferences.


Ion flux and the function of endosomes and lysosomes: pH is just the start: the flux of ions across endosomal membranes influences endosome function not only through regulation of the luminal pH.
The ionic nature of endosomes varies considerably in character along the endocytic pathway. Counter-ion flux across the limiting membrane of endosomes has long been considered essential for full acidification and normal endosome/lysosomal function. The proximal functions of luminal ions, however, have been difficult to assess. The recent development of transgenic mice carrying mutations in the intracellular chloride channels (ClCs) has provided a tool to uncouple Cl(-) influx from endosomal acidification. Intriguingly, many of the defects of the endo-lysomal system attributed to aberrant pH persist in the Cl(-)-deficient mice implying a direct regulatory role for Cl(-) influx in endosome function. These observations may begin to explain the abundance of endosomal ion transporters, including ClCs, sodium-proton exchangers, two-pore channels and mucolipins, that have been localized to endo-lysosomes, and the extensive changes in luminal ion composition therein. In this review, we summarize what is known regarding the mediators of endosomal ion flux, and discuss the implications of changing ionic content on endo-lysosomal function.


Impact of a long photoperiod during lactation on immune status of piglets.
The effects of a long photoperiod treatment around parturition and throughout lactation on immune status of piglets were studied. Sows were assigned to 2 light regimens: i) standard short photoperiod (SP, n = 17), 8 h of daily light from d 112 of gestation until d 23 of lactation; and ii) long photoperiod (LP, n = 17), 23 h of daily light from d 112 of gestation to d 4 of lactation and 16 h thereafter. In front of the crates, under the side heat lamps and behind the sow, light intensities were 59 ± 5, 109 ± 6, and 44 ± 6 lx, respectively. On d 15 of lactation and at weaning (d 23), 2 piglets of similar BW per litter were selected and immunized intramuscularly with ovalbumin (OVA). Blood samples (5 mL serum and 10 mL whole blood) were taken at d 15 and d 23 of lactation, and at d 30, 37, and 44 of age after weaning to evaluate the antibody response to OVA and measure phagocytosis, lymphocyte proliferative response, and different circulating blood lymphocyte populations of piglets. Results showed that phagocytosis was increased in piglets submitted to LP (P < 0.05). A treatment × time interaction (P < 0.001) indicated that SP piglets developed a better IgG response to OVA than LP piglets. The percentage of B lymphocytes was also increased (P = 0.02) in SP piglets compared with piglets exposed to LP during lactation; the lymphocyte response to OVA tended to be enhanced (P = 0.07) over time in SP piglets. Different subpopulations of CD8+ lymphocytes were markedly increased in SP piglets at 23 d of age compared with piglets exposed to LP (treatment × time: P < 0.05). These results suggest that exposure of piglets to LP during lactation seems to reduce the capacity of piglets to develop a strong immune response to novel antigens. This may have important consequences on the ability of piglets to resist an infection after weaning.


A new proton conductive liquid with no ions: pseudo-protic ionic liquids.
Liquids with no ions! Raman analysis and quantum calculations suggest that electrically neutral molecular species predominantly exist in an N-methylimidazole and acetic acid equimolar mixture, and that ionic species are rather minor. Nevertheless, the mixture has significant ionic conductivity, and shows "good ionic" or "superionic" behavior (see figure). It may be suitable to call such liquids "pseudo-ionic liquids" rather than "ionic liquids".


Postthaw viability of the inner cell mass of in vitro-matured/in vitro-fertilized bovine embryos frozen in various cryoprotectants.
A study was conducted to examine the viability of inner cell mass (ICM) cells of frozen-thawed in vitro-matured (IVM)/in vitro-fertilized (IVF)-derived embryos using various cryoprotectants. Expanded blastocysts were frozen and thawed in 1.4 M glycerol with 0.25 M sucrose (GL), 1.6 M propylene glycol (PG), 1.8 M ethylene glycol (EG), or 1.3 M ethylene glycol monomethyl ether (EME) as cryoprotectants using a one-step method. After thawing, the embryos were cocultured for 24 h with cumulus cells in TCM199. Embryos which were viable after thawing and developed beyond the blastocyst stage were treated by immunosurgery and differential fluorochrome staining for ICM cell counts. Overall, there were no significant differences in the development to blastocyst stage after 24 h culture in each cryoprotectant (P < 0.05, chi 2 analysis). The viability of ICM cells of frozen-thawed embryos with each cryoprotectant was lower (GL, 72.7%; PG, 67.8%; EG, 77.5%; EME, 74.7%) than that of unfrozen embryos (84.4%). In the case of PG as a cryoprotectant, viability of ICM cells was significantly lower than that of unfrozen embryos (P < 0.05, ANOVA analysis). Our results suggest that the viability of ICM cells of frozen-thawed bovine embryos tend to be lower than that of unfrozen embryos irrespective of the cryoprotectants used. PG was significantly more toxic to the ICM cells compared with the other cryoprotectants.


Direct and indirect effects of plant diversity and phenoxy herbicide application on the development and reproduction of a polyphagous herbivore.
Widespread application of synthetic pesticides and loss of plant diversity are regarded as significant drivers of current global change. The effects of such phenomena on insect performance have been extensively studied separately, yet the interactions of these two drivers have been poorly explored. Here, we subjected the polyphagous grasshopper Pseudochorthippus parallelus (Zetterstedt, 1821) to a full-lifecycle field experiment with 50 cages containing experimental plant communities differing in grass species richness (2 vs. 8 grass species), half of them treated with a phenoxy herbicide commonly employed to control broadleaf plants in grasslands. We measured plant elemental content as a proxy for plant physiology, and a wide range of insect traits in both female and male grasshoppers. In females, grass diversity increased herbivory, insect nitrogen content and egg load, while herbicide reduced herbivory but increased the number of offspring, likely mediated by altered plant community composition. In males, grass diversity also increased herbivory, had positive effects on fat body, muscle volume and lifespan, and negative effects on body mass. Herbicide negatively affected herbivory in both females and males. Overall, plant diversity and herbicides may shift resource allocation in generalist terrestrial insect herbivores, indicating complex and unexpected effects of human-induced environmental change.


Maple syrup urine disease: it has come a long way.
Maple syrup urine disease (MSUD) was first described in 1954 by Menkes et al. as a progressive neurologic degenerative disorder. In 1960, Dancis et al. established that the metabolic block in MSUD is at the decarboxylation of branched-chain alpha-ketoacids derived from leucine, isoleucine, and valine. The multienzyme complex affected in MSUD, the mitrochondrial branched-chain alpha-ketoacid (BCKD) dehydrogenase complex was purified in 1978 to homogeneity in Reed's laboratory. This led to the later cloning of cDNAs and genes for subunits of the human BCKD complex. Genetic heterogeneity in MSUD is now explained by the various mutations that occur in the E1 alpha, E1 beta, E2, and E3 loci of the BCKD complex. Recently, we found that bacterial chaperonins GroEL and GroES promote folding and assembly of E1 decarboxylase component of the BCKD complex in Escherichia coli. Pulse-chase labeling in this system showed that a subset of E1 alpha mutations, notably the homozygous Y393N-alpha in Mennonite MSUD patients, impedes the assembly of the mutant E1 alpha subunit with normal E1 beta. The assembly defect is associated with a rapid degradation of the normal E1 beta subunit in MSUD cells. Retrovirus-mediated transduction of lymphoblasts from a Mennonite MSUD patient with a normal E1 alpha cDNA resulted in a complete restoration of BCKD activity. This was accompanied by a stabilization of the normal E1 beta subunit through assembly with recombinant E1 alpha. The results demonstrated the feasibility of stable correction of E1 alpha-deficient (type IA) MSUD and provided a basis for the development of gene therapy.


Health care resource utilization and costs among individuals with vs without Huntington disease in a US population.
BACKGROUND: Quantifying the extent of health care resource utilization (HCRU) and costs associated with Huntington disease (HD) is vital for providers, decisionmakers, and payers to understand unmet treatment needs and to ensure limited resources can be used to benefit the maximum number of people with HD. OBJECTIVE: To quantify HCRU and costs for people with HD, overall and by disease stage, and compare these with non-HD controls. METHODS: This was a retrospective cohort study using administrative claims data from the IBM MarketScan Commercial, Multi-State Medicaid, and Medicare Supplemental Databases from January 1, 2009, to December 31, 2018. People with an HD claim between January 1, 2010, and December 31, 2017, were selected for this analysis and matched with non-HD controls for comparison. The HD cohort and the non-HD controls were exact matched on their follow-up duration and propensity score matched 1:4 to create the final analytical cohort. Index date was the first HD diagnosis for the HD cohort (proxy index date assigned to controls), and all individuals were required to have continuous enrollment for 12 or more months pre-index (baseline) and 3 or more months post-index. Proportions of all-cause HCRU (ie, outpatient visits, inpatient visits, emergency department visits, pharmacy fills, radiology visits, and physical/occupational therapy visits) in the 6-months post-index and HCRU counts and costs per patient per month (PPPM) over the entire follow-up were calculated for each cohort. RESULTS: A total of 2,473 individuals with HD and 9,522 matched non-HD controls were identified. HCRU in 6 months post-index was significantly greater in people with HD compared with non-HD controls for all health care service categories; P < 0.0001. The mean number of HCRU PPPM for all measured healthcare services was significantly higher in people with HD compared with non-HD controls (P < 0.001). Mean total costs (2018 USD PPPM) for the HD cohort ($2,260 [SD = $4,682]) were twice the total costs in the non-HD cohort ($1,056 [SD = $3,078]) (P < 0.0001) and were highest across all disease stages. CONCLUSIONS: This study provides current comprehensive HCRU and cost estimates in individuals with HD relative to those without the disease, thus demonstrating the high economic burden imposed by HD. DISCLOSURES: Dr Ta: Employment with Genentech (at time of study) and stock options with Roche; Dr To: Employment and stock options/dividends with Genentech; Dr Patel: Employment and stock options with Roche/Genentech; Dr Fuller: Employment with CHDI Management/CHDI Foundation; Mr Surinach: Employment with Genesis Research (which receives consulting fees from Genentech/Roche); Dr Abbass: Employment and stock options with Genentech; Dr Exuzides: Employment and stock options with Roche/Genentech; and Ms Luo: Employment with CHDI Management/CHDI Foundation. This study was funded by Genentech Inc. The authors thank Greg Rowe of Chrysalis Medical Communications for providing medical writing support, which was funded by F. Hoffmann-La Roche Ltd, in accordance with Good Publication Practice (GPP3) guidelines (http://www.ismpp.org/gpp3).


Stability and iron coordination in DNA adducts of Anthracycline based anti-cancer drugs.
There is evidence that the interaction of the α-ketol group of the Doxorubicin and Epirubicin anti-cancer drugs with Fe(III) generates hydroxyl radicals under aerobic conditions, causing cardiotoxicity in patients. Considering that the formation of DNA adducts is one of the main targets of Anthracycline drugs, we have in the present study characterized several [Anthracycline-DNA]Fe(III) complexes with respect to their stability and Fe(III) coordination, by means of MD simulations. Iron is found to coordinate well to the drugs containing an α-ketol group, this being the only group of the drug that binds to the metal. The complexes containing an α-ketol group, [Doxorubicin-DNA]Fe(III) and [Epirubicin-DNA]Fe(III), thus show greater stability than those not containing it, i.e., [Daunorubicin-DNA]Fe(III), [Idarubicin-DNA]Fe(III) and [5-Imino-Daunorubicin]Fe(III). Metal attachment to the α-ketol group is furthermore facilitated by the phosphate groups of DNA. The coordination to iron in the [Doxorubicin-DNA]Fe(III) system is smaller than that found for the [Epirubicin-DNA]Fe(III) system, and the corresponding number of coordinating waters in the former is larger than in the latter. This may in turn result in higher hydroxyl radical production, thus explaining the increased cardiotoxicity noted for Doxorubicin.


Lung function in military oxygen divers: a longitudinal study.
It is increasingly recognized that professional diving may elicit adverse long-term effects on the lungs, but conflicting results have been reported from distinct diving cohorts. This study reports the longitudinal change in lung function in professional divers who employ closed-circuit oxygen rebreathing apparatuses. All oxygen divers who attended the German Naval Medical Institute between 1994 and 1999 for regular medicals underwent spirometry and were entered if they had at least two follow-up examinations. Forced expiratory flows and volumes at baseline and at maximum follow up were compared. There were 39 divers who presented at least 3 times during a median period of 5.8 (2.7-8) yr. Forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC) amounted to 4.86 +/- 0.62 L and 5.89 +/- 0.67 L at baseline, and 4.83 +/- 0.64 L and 5.87 +/- 0.69 L at maximum follow up, respectively. The change over time was statistically not significant. Substantial exposure to elevated oxygen partial pressure while diving is not associated with an accelerated decline in lung function. Factors other than hyperoxia (e.g., venous gas microemboli and altered breathing gas characteristics) may account for the long-term effects that have been found in professional divers.


Analysis and comparison of clinicohematological parameters and molecular and cytogenetic response of two Bcr/Abl fusion transcripts.
Different forms of p210 are produced by alternative splicing, namely b2a2 and b3a2. There have been many contrasting data establishing a relationship between the two Bcr/Abl transcripts and platelet counts and also response to treatment. However, the data published to date have been on a small group of patients. The aim of the present study was to determine whether there was any difference between clinical and hematological parameters at diagnosis between the two Bcr/Abl fusion transcripts in our population, and whether the two transcripts responded differently or similarly to imatinib treatment. RT-PCR was performed in 202 cases for detection of Bcr/Abl transcripts in newly diagnosed chronic myelogenous leukemia cases in one year. The two transcripts were compared and correlated with clinical, hematological and FISH data and with response to treatment. A total of 138 cases were of b3a2 and 64 were of b2a2 transcript. There was no correlation between the hematological parameters and the type of transcript. There was a significant association of blast crisis with b2a2, especially with myeloid blast crisis. When compared to FISH results, 10% of b3a2 were found to have a significant association with 5'Abl deletion as compared to 3% of b2a2. On analyzing the therapeutic response, we did not find any difference between the two transcripts. In conclusion, our findings confirm that the b3a2 type transcript is not significantly associated with thrombocytosis, that the short transcript, b2a2, occurs with acute phase, i.e., blast crisis, and that there is no difference in treatment response between the two transcripts. However, further studies are required to understand the molecular pathways involved in the Bcr/Abl mechanism.


Joint recommendations on management of anaemia in patients with gastrointestinal bleeding in Hong Kong.
The demand for blood products continues to grow in an unsustainable manner in Hong Kong. While anaemia associated with gastrointestinal bleeding (GIB) is the leading indication for transfusion, there is no local recommendation regarding best practices for transfusion. We aimed to provide evidence-based recommendations regarding management of anaemia in patients with acute and chronic GIB. We reviewed all original papers, meta-analyses, systematic reviews, or guidelines that were available in PubMed. For acute GIB, a restrictive transfusion strategy, targeting a haemoglobin threshold of 7 to 8 g/dL, should be adopted because overtransfusion is associated with significantly higher all-cause mortality and re-bleeding. A liberal transfusion strategy should only be considered in patients with co-existing symptomatic coronary artery disease, targeting a haemoglobin threshold of 9 to 10 g/dL. When acute GIB settles, patients should be prescribed iron supplements if iron deficiency is present. For chronic GIB, iron stores should be replenished aggressively via iron supplementation before consideration of blood transfusion, except in patients with symptoms of severe anaemia. Oral iron replacement is the preferred first-line therapy, while intravenous iron is indicated for patients with inflammatory bowel disease, poor response or poor tolerability to oral iron, and in whom a rapid correction of iron deficit is preferred. Intravenous iron is underutilised and the risk of anaphylactic reaction to current preparations is extremely low. These recommendations are provided to local clinicians to facilitate judicious and appropriate use of red cell products and iron replacement therapy in patients with GIB.


A mutation study of sperm head shape and motility in the mouse: lessons for the clinic.
Mouse mutants that show effects on sperm head shape, the sperm tail (flagellum), and motility were analysed in a systematic way. This was achieved by grouping mutations in the following classes: manchette, acrosome, Sertoli cell contact, chromatin remodelling, and mutations involved in complex regulations such as protein (de)phosphorylation and RNA stability, and flagellum/motility mutations. For all mutant phenotypes, flagellum function (motility) was affected. Head shape, including the nucleus, was also affected in spermatozoa of most mouse models, though with considerable variation. For the mutants that were categorized in the flagellum/motility group, generally normal head shapes were found, even when the flagellum did not develop or only poorly so. Most mutants are sterile, an occasional one semi-sterile. For completeness, the influence of the sex chromosomes on sperm phenotype is included. Functionally, the genes involved can be categorized as regulators of spermiogenesis. When extrapolating these data to human sperm samples, in vivo selection for motility would be the tool for weeding out the products of suboptimal spermiogenesis and epididymal sperm maturation. The striking dependency of motility on proper sperm head development is not easy to understand, but likely is of evolutionary benefit. Also, sperm competition after mating can never act against the long-term multi-generation interest of genetic integrity. Hence, it is plausible to suggest that short-term haplophase fitness i.e., motility, is developmentally integrated with proper nucleus maturation, including genetic integrity to protect multi-generation fitness. We hypothesize that, when the prime defect is in flagellum formation, apparently a feedback loop was not necessary as head morphogenesis in these mutants is mostly normal. Extrapolating to human-assisted reproductive techniques practice, this analysis would supply the arguments for the development of tools to select for motility as a continuous (non-discrete) parameter.


Structure of avian orthoreovirus virion by electron cryomicroscopy and image reconstruction.
Among members of the genus Orthoreovirus, family Reoviridae, a group of non-enveloped viruses with genomes comprising ten segments of double-stranded RNA, only the "non-fusogenic" mammalian orthoreoviruses (MRVs) have been studied to date by electron cryomicroscopy and three-dimensional image reconstruction. In addition to MRVs, this genus comprises other species that induce syncytium formation in cultured cells, a property shared with members of the related genus Aquareovirus. To augment studies of these "fusogenic" orthoreoviruses, we used electron cryomicroscopy and image reconstruction to analyze the virions of a fusogenic avian orthoreovirus (ARV). The structure of the ARV virion, determined from data at an effective resolution of 14.6 A, showed strong similarities to that of MRVs. Of particular note, the ARV virion has its pentameric lambda-class core turret protein in a closed conformation as in MRVs, not in a more open conformation as reported for aquareovirus. Similarly, the ARV virion contains 150 copies of its monomeric sigma-class core-nodule protein as in MRVs, not 120 copies as reported for aquareovirus. On the other hand, unlike that of MRVs, the ARV virion lacks "hub-and-spokes" complexes within the solvent channels at sites of local sixfold symmetry in the incomplete T=13l outer capsid. In MRVs, these complexes are formed by C-terminal sequences in the trimeric mu-class outer-capsid protein, sequences that are genetically missing from the homologous protein of ARVs. The channel structures and C-terminal sequences of the homologous outer-capsid protein are also genetically missing from aquareoviruses. Overall, the results place ARVs between MRVs and aquareoviruses with respect to the highlighted features.


Effects of histamine and related compounds on thyrotropin secretion in rats.
The effects of histamine (HA) and related compounds on thyrotropin-releasing hormone (TRH) and thyrotropin (TSH) secretion in rats were studied. Histidine (1.0 g/kg), HA (5.0 mg/kg) or histamine antagonists mepyramine (MP) (100 mg/kg) or famotidine (FA) (5.0 mg/kg) were injected intraperitoneally, and the rats were decapitated at various intervals after the injection. The hypothalamic immunoreactive TRH (ir-TRH) content increased significantly after histidine or HA injection, decreased significantly after FA injection, but was not changed by MP. The plasma ir-TRH concentration did not change significantly after injection of these drugs. The plasma TSH levels decreased significantly in a dose-related manner after histidine or HA injection and increased significantly in a dose-related manner after FA injection. The plasma thyroid hormone levels showed no changes. In the FA-pretreated group, the inhibitory effect of histidine or HA on TSH levels was prevented, but not in the MP-pretreated group. The plasma ir-TRH and TSH responses to cold were inhibited by histidine or HA and enhanced by FA. The plasma TSH response to TRH was inhibited by histidine or HA and enhanced by FA. The inactivation of TRH immunoreactivity by hypothalamus or plasma in vitro after histidine, HA, MP or FA was not different from that of the control. These findings suggest that histamine may act both on the hypothalamus and the pituitary to inhibit TRH and TSH release, and that its effects may be mediated via H2-receptor.


Gestation length in farmed reindeer.
Reindeer (Rangifer tarandus tarundus) are the only cervids indigenous to the arctic environment. In Alaska, reindeer are a recognized agricultural species and an economic mainstay for many native populations. Traditionally raised in extensive free-ranging systems, a recent trend toward intensive farming requires a more in-depth knowledge of reproductive management. Reported gestation length in reindeer varies, ranging from 198 to 229 d in studies performed at the University of Alaska Fairbanks. A switchback study that manipulated only breeding date demonstrated a mean increase in gestation length of 8.5 d among females bred early in the season. The negative correlation between conception date and gestation length is consistent with reindeer research at other locations and reports of variable gestation length in a growing number of domestic and non-domestic species. This paper reviews the phenomenon in reindeer and discusses some of the factors known to affect gestation length as well as possible areas for future research.


PAR1-type thrombin receptor stimulates migration and matrix adhesion of human colon carcinoma cells by a PKCepsilon-dependent mechanism.
The proteinase-activated receptor1 (PAR1) was characterized as a functional receptor for thrombin in cells from different tumor entities. In colon carcinoma, its function has to be defined. In this study we demonstrate that the PAR1-selective agonist peptide TFLLRN induced activation of protein kinase C isoenzymes alpha and epsilon in human HT-29 colon carcinoma cells expressing PAR1 endogeneously. On the cellular level, TFLLRN and thrombin prompted HT-29 cell migration and matrix adhesion by a PKCepsilon-dependent mechanism as concluded because of the inhibition of PAR1-mediated effects by the PKC inhibitor bisindolylmaleimide I and the PKCepsilon translocation inhibitory peptide EAVSLKPT but not by the PKC inhibitor Gö 6976. In addition, blockade of PAR1 by RWJ 56110, a selective PAR1 antagonist, fully abolished the effect of thrombin on HT-29 cell migration and adhesion. Therefore, PAR1 seems to be the responsible receptor for thrombin-induced migration and adhesion of human colon carcinoma cells including PKCepsilon as an essential signal transducer.


Conformational stability of OXA-51 β-lactamase explains its role in carbapenem resistance of Acinetobacter baumannii.
Acinetobacter baumannii, an important nosocomial pathogen, is increasingly becoming resistant to antibiotics including recent β-lactam like imipenem. Production of different types of β-lactamases is one of the major resistance mechanisms which bacteria adapt. We recently reported the presence of a β-lactamase, OXA-51, in clinical strains of A. baumannii in ICUs of our hospital. This study is an attempt to understand the structure-function relationship of purified OXA-51 in carbapenem resistance in A. baumannii. The OXA-51 was cloned, expressed in E. coli Bl-21(DE3) and further purified. The in vitro enzyme activity of purified OXA-51 was confirmed by two independent techniques; in-gel assay and spectrophotometric method using nitrocefin. Further in vivo effect of OXA-51 was followed by transmission electron microscopy of bacterium. Biophysical and biochemical investigations of OXA-51 were done using LC-MS/MS, UV-Vis absorption, fluorescence, circular dichroic spectroscopy and isothermal calorimetry. Native OXA-51 was characterized as 30.6 kDa, pI 8.43 with no disulphide bonds and comprising of 30% α-helix, 27% β-sheet. Secondary structure of OXA-51 was significantly unchanged in broad pH (4-10) and temperature (30-60 °C) range with only local alterations at tertiary structural level. Interestingly, enzymatic activity up to 75% was retained under above conditions. Hydrolysis of imipenem by OXA-51 (k(m),1 μM) was found to be thermodynamically favourable. In the presence of imipenem, morphology of sensitive strain of A. baumannii was drastically changed, while OXA-51-transformed sensitive strain retained the stable coccobacillus shape, which demonstrates that imipenem is able to kill sensitive strain but is unable to do so in OXA-51-transformed strain. Hence the production of pH- and temperature-stable OXA-51 appears to be a major determinant in the resistance mechanisms adopted by A. baumannii in order to evade even the latest β-lactams, imipenem. It can be concluded from the study that OXA-51 plays a vital role in the survival of the pathogen under stress conditions and thus poses a major threat.


Data Driven Computational Design and Experimental Validation of Drugs for Accelerated Mitigation of Pandemic-like Scenarios.
Emerging pathogens are a historic threat to public health and economic stability. Current trial-and-error approaches to identify new therapeutics are often ineffective due to their inefficient exploration of the enormous small molecule design space. Here, we present a data-driven computational framework composed of hybrid evolutionary algorithms for evolving functional groups on existing drugs to improve their binding affinity toward the main protease (Mpro) of SARS-CoV-2. We show that combinations of functional groups and sites are critical to design drugs with improved binding affinity, which can be easily achieved using our framework by exploring a fraction of the available search space. Atomistic simulations and experimental validation elucidate that enhanced and prolonged interactions between functionalized drugs and Mpro residues result in their improved therapeutic value over that of the parental compound. Overall, this novel framework is extremely flexible and has the potential to rapidly design inhibitors for any protein with available crystal structures.


Electromagnetic navigation-guided preoperative localization: the learning curve analysis.
The electromagnetic navigation bronchoscopy (ENB) was increasingly used to mark small pulmonary nodules (PNs) for video-assisted thoracic surgery (VATS) resection due to high effectiveness and low risk. However, no study reports the learning curve of ENB-guided preoperative localization. In the study, we aimed to describe the learning curve of ENB-guided preoperative PNs localization initially. Consecutive PNs cases that underwent ENB localizations between October 2018 and October 2019 by the same surgeon in our center were included in the study. The cumulative sum (CUSUM) method was used to analyze the learning curve of ENB localization. A total of 89 ENB localization from 64 patients were included in this study. The learning curve was divided into 3 phases: Phase I (the initial 11 cases), Phase II (the 12th to the 47th cases), and Phase III (the 47th to the 89th cases). The success rate of ENB localization has increased with the accumulation of operational experience in 3 phases (72.73%, 91.67%, and 97.62%, P=0.049). The distance from the ENB guide wire tip to the center of the lesion in Phase I was significantly longer than those in Phase II and Phase III (2.46±1.76 vs. 1.36±0.94 and 1.47±0.97 cm, P=0.014 and 0.027, respectively). Sex, bronchus sign, and learning curve phase were independent risk factors influencing operative time (OT) (OR =8.187, 18.847, and 13.920, respectively). The technical competency, which is indicated by higher success rate, localization accuracy, and shorter OT, for ENB-guided preoperative PNs localization was achieved at the 47th operation.


Impact of Previous Conventional Cardiac Surgery on the Clinical Outcomes After Heart Transplantation.
The impact of the type, purpose, and timing of prior surgery on heart transplantation (HT) remains unclear. This study investigated the influence of conventional cardiac surgery (PCCS) on HT outcomes. This study analyzed HTs performed between 1999 and 2019 at a single institution. Patients were categorized into two groups: those with and without PCCS. Short-term outcomes, including post-transplant complications and mortality rates, were evaluated. Cox proportional and Kaplan-Meier survival analyses were used to identify risk factors for mortality and assess long-term survival, respectively. Of 368 patients, 29% had PCCS. Patients with PCCS had a higher incidence of post-transplant complications. The in-hospital and 1 year mortality rates were higher in the PCCS group. PCCS and cardiopulmonary bypass time were significant risk factors for 1 year mortality (hazard ratios = 2.485 and 1.005, respectively). The long-term survival rates were lower in the PCCS group, particularly in the first year. In sub-analysis, patients with ischemic cardiomyopathy and PCCS had the poorest outcomes. The era of surgery and timing of PCCS in relation to HT did not significantly impact outcomes. In conclusion, PCCS worsen the HT outcomes, especially in patients with ischemic etiology. However, the timing of PCCS and era of HT did not significantly affect this concern.


Two novel compounds from Paeonia suffructicosa.
A new hexacyclic triterpenoid, mudanpinoic acid A (1), and a new gallic acid glycoside, mudanoside B (2), along with nine known compounds--benzoic acid, resacetophenone, paeoniflorigenone, beta-sitosterol, betulinic acid, oleanoic acid, quercetin, beta-sitosterol-beta-D-glucoside, and trans-caffeic acid stearyl ester-were isolated from the dried root cortex of Paeonia suffruticosa. The structures of the novel compounds were elucidated on the basis of spectral methods, and that of compound 1 was confirmed by X-ray crystallographic analysis.


Influence of exposure and vertical transmission of HIV-1 on the neuropsychomotor development in children.
Exposure to human immunodeficiency virus (HIV)-1 during pregnancy is a major risk factor for neurodevelopmental delay and deleterious effects in children. However, limited information about these conditions exists in poor geographical areas in Brazil. Prevention of vertical transmission of HIV-1 is dealt differently in different regions of the country and in poorer areas it is more difficult to evaluate the impact of the prevention methods. The outcomes of the exposure to HIV-1 and the impact of vertical HIV-1 transmission on neuropsychomotor development was evaluated for the first time in children born to HIV-infected mothers in the North region of Brazil, where the majority of the population has poor access to health services. Sixty children born to HIV-1-infected mothers (case group) and 58 born to non-infected mothers (control group) were followed for the first 12 months of life in a prospective case-control study. Neuropsychomotor development was assessed using the Denver II test. Suspected neuropsychomotor developmental delays were more frequent in the case group (33.3%), namely in language (38.9%) and gross motor skills (27.8%). These delays were reversed in most children after 12 months of life due to therapeutic intervention. The delays were not reversed in three children, all of whom belonged to the case group. Only one of these was infected with HIV-1, and this child had the poorest neuropsychomotor outcomes. Maternal HIV-1 infection negatively affected the neuropsychomotor development in children, although other factors may have played a role.


Multiple intracellular pathways interfere with the activation of a CPP32-like protease induced by serum deprivation of AKR-2B cells.
As previously described, confluent AKR-2B fibroblasts rapidly disintegrate upon removal of serum. Platelet-derived growth factor isoforms AB or BB (PDGF-AB, -BB) added immediately after serum deprivation caused complete survival of the cells without initiating proliferation (Simm et al., 1994, J. Cell. Physiol. 160, 295). Here the role of cAMP as a protective agent was investigated by using forskolin or 8-Br-cAMP. Both reagents afforded high cellular protection. The phorbolester TPA, an activator of protein kinase C isoforms, also exerted a high protection against cell death (ED50 = 7 nM). Unexpectedly colchicine (ED50 = 1.5 microM) an inhibitor of tubulin polymerization also protected cells from death. The protective effects of PDGF-BB and TPA were dependent on protein synthesis as indicated by their complete suppression by cycloheximide (CHx). Surprisingly, forskolin and 8-Br-cAMP remained effective even in the presence of CHx. Detailed studies of several signalling pathways were performed. These investigations showed no interference between PDGF-BB and cAMP-dependent pathways at the early stage of signal transduction. As previously described, the ICE-like protease inhibitor tyr-val-ala-asp-chloromethylketone (YVAD-cmk) protected cells from death (Simm et al., 1997, J. Cell Sci. 110, 819-828). As shown here, a substantial protection was also achieved by the addition of two other caspase inhibitors: asp-glu-val-asp-aldehyde (DEVD-cho; ED50 = 100 microM) and benzoylcarbonyl-asp-glu-val-asp-chloromethylketone (Z-DEVD-cmk; ED50 = 100 microM). The activity of caspases was studied using either tyr-val-ala-asp-aminomethylcoumarine (YVAD-amc) or asp-glu-val-asp-aminomethylcoumarine (DEVD-amc) as substrates. There was no activation of a YVADase, whereas as pronounced increase in DEVDase activity was found with a maximum 3 h after serum removal. Cross competition experiments in vitro showed that the latter activity is inhibited also by low concentrations of YVAD-cmk (300-600 nM), suggesting that both inhibitors inactivated the same target protease. Remarkably all tested protective reagents lead to an inhibition of the DEVDase activity in intact cells. Since these reagents act via distinct intracellular pathways, the existence of a regulatory element upstream of the DEVDase is proposed which integrates signals from a variety of pathways.


Breast feeding: success or failure.
Awareness of the important role of breast feeding in child health and the reproductive cycle, which is well documented elsewhere, necessitates an examination of the changing practice of breast feeding. This paper reviews these changing practices both in Western and developing societies, examines the problems which lead to lactation failure, and looks at factors related to success in breast feeding. In the light of the need for practical help for successful breast feeding. the present system of support both in hospital and at home is then discussed. An alternative system of structured home support for post-natal women, which has been shown to increase the success rate of breast feeding, is outlined. The way in which this system acts is discussed and the implications for both mothers and health staff are considered.


Enhancing cellulose dissolution in ionic liquid by solid acid addition.
It's challenging to dissolve natural cellulose in most solvents due to its highly ordered crystalline structure. In this paper, we developed an efficient cellulose dissolution system which incorporates solid acid (SA) with 1-butyl-3-methylimidizolium chloride (BmimCl). The results showed that addition of solid acid both Amberlyst® 15 and CsxH3-xPW12O40 could significantly enhance cellulose dissolution in BmimCl, which attributed to the synergistic action of free hydrogen proton from SA and chloride anion in BmimCl on hydroxyl groups of cellulose, and DMF as co-solvent also could facilitate cellulose dissolution in SA/BmimCl. In contrast to BmimCl system, the SA/BmimCl system for cellulose dissolution achieves better efficiency at mild treatment conditions and facile recovery of solvents. In addition, characterization of the regenerated celluloses showed that SA/BmimCl is a non-derivatizing solvent for cellulose, which helps achieve complete dissolution on crystalline cellulose. Attributed to its low cost and environmentally friendliness for biomass processing, SA/BmimCl systems is a promising and effective solvent system.


The electronic structures of Co and Ni tetraazaannulenes.
We compare the electronic structure of two metal-centered tetramethyldibenzo-tetraazaannulene (TMTAA) macrocyclic complex molecules: 5,7,12,14- tetramethyl-2,3:9,10-dibenzo[b,i]-1,4,8,11-tetraazacyclotetradecine nickel (II) and 5,7,12,14-tetramethyl-2,3:9,10-dibenzo[b,i]-1,4,8,11-tetraazacyclotetradecine cobalt (II). The experimental gap between the highest occupied molecular orbital to the lowest unoccupied molecular orbital for both molecules, obtained from combined ultraviolet photoemission and inverse photoemission studies, is close to the value of 6.6 eV expected from simple model calculations, but with the Fermi level placed closer to the lowest unoccupied molecular orbital. While both the Co(II) (s = 1/2) and Ni(II) (s = 0) TMTAA molecular electronic structures are very similar, the Ni(II) adopts a high-symmetry molecular configuration upon adsorption, with a strong preferential orientation.


Clinical and immunologic analyses of 103 patients with common variable immunodeficiency.
Common variable immunodeficiency (CVI) or hypogammaglobulinemia is a heterogeneous primary immunodeficiency disease in which B cells produce little or no antibody. Since the disease is relatively rare and the spectrum of associated illnesses is broad, patients are given care by a variety of specialists. Thus it has been difficult to determine the incidence of specific complications. In these studies we analyzed 103 consecutively referred CVI patients of age range 3-71 years (average, 29 years) who were followed for a period of 1-13 years (total of 750 patient years). The average serum IgG was 174.4 mg/dl for untreated patients and 301 mg/dl for patients treated with intramuscular immunoglobulin at the time of the first visit. The average IgA was 14.5, and the average IgM was 80.7, with no difference between or after immunoglobulin treatment. About one-half of the patients had T-cell dysfunction, but lymphocyte stimulation responses were inversely related to age, which implies worsened T-cell immunity with age. Serum IgG and IgA levels were found to be statistically associated (P = 0.008), and serum IgG was related to lymphocyte stimulation with concanavalin A (P = 0.01). By 1986, 79 patients were alive, 23 had died, and 1 could not be located. Recurrent bacterial illnesses were common to all patients, and 22% had developed chronic lung disease, 22% autoimmune disease, 15% cancer, 13% hepatitis, and 9% malabsorption. Autoimmune disease was more common in females, and cancer was more likely to develop in the fifth and sixth decades. In 11% of the group, other family members were found to be immunodeficient (hypogammaglobulinemic or IgA deficient). Nine patients died of respiratory insufficiency (with or without other complications), and seven patients died of cancer. These data provide valuable information about the immunologic abnormalities and the spectrum and frequency of illnesses associated with hypogammaglobulinemia.


Percutaneous vertebroplasty in Moroccan patients with vertebral compression fractures.
Incidence of vertebral compression fractures (VCFs) is increasing due to the increase in human life expectancy and prevalence of osteoporosis. Percutaneous vertebroplasty (PVP) has become a widely used alternative treatment for symptomatic VCFs refractory to medical therapy. It is a minimally invasive technique involving injection most frequently of polymethylmethacrylate (PMMA) directly into the vertebral body through the pedicles. Percutaneous vertebroplasty yields good results in terms of early vertebral stabilization and prompt pain relief. This study describes the experience of the Department of Neurosurgery at Ibn Sina University Hospital (Rabat, Morocco) and assesses short and long term clinical and radiological outcomes and complications of vertebroplasty in a retrospective assessment of 36 vertebral compression fractures in 35 patients (16 men and 19 women subjects) who had been treated with vertebroplasty from November 2006 to December 2014.


Identification of genetic mutations associated with attenuation and changes in tropism of Urabe mumps virus.
Although several effective mumps virus vaccines have been developed, almost nothing is known about the genetic changes responsible for loss of virulence. One vaccine, Urabe AM9, was withdrawn from the market because of insufficient attenuation. The vaccine was found to contain a mixture of viruses that could be distinguished based on the sequence of the hemagglutinin-neuraminidase gene (HN). Viruses containing lysine at HN amino acid position 335 were isolated from cases of post-vaccination parotitis or meningitis whereas viruses containing glutamic acid at this position were not associated with post-vaccination disease. Using a rat based model of mumps neurovirulence, we demonstrate that this latter virus is significantly attenuated compared to a virus isolated from a patient with post-vaccination meningitis. Complete sequence analysis of the genomes of the two viruses identified sixteen genetic differences, some or all of which must be responsible for differences in virulence. These same genetic differences also account for changes in tropism in cell culture.


A Noncovalent π-Stacked Porous Organic Molecular Framework for Selective Separation of Aromatics and Cyclic Aliphatics.
Selective separation using porous adsorbents is an energy-efficient alternative to traditional separation techniques. Stacked porous organic molecular frameworks (POMFs) capable of noncovalent π⋅⋅⋅π interactions are emerging as a new kind of adsorbents that facilitate green separation. Here we report a robust porous molecular crystal (TAPM-1), which is stabilized by multiple intermolecular π⋅⋅⋅π interactions. With its long-range π-stacking, TAPM-1 has excellent hydrophobicity, thermostability, recyclability, and high selectivity for aromatics over the corresponding cyclic aliphatics. This enables TAPM-1 to serve as the stationary phase in the high-resolution gas chromatographic separation of benzene and cyclohexane or toluene and methylcyclohexane.


Characterisation of a monoclonal antibody detecting Atlantic salmon endothelial and red blood cells, and its association with the infectious salmon anaemia virus cell receptor.
Endothelial cells (ECs) line the luminal surfaces of the cardiovascular system and play an important role in cardiovascular functions such as regulation of haemostasis and vasomotor tone. A number of fish and mammalian viruses target these cells in the course of their infection. Infectious salmon anaemia virus (ISAV) attacks ECs and red blood cells (RBCs) of farmed Atlantic salmon (Salmo salar L.), producing the severe disease of infectious salmon anaemia (ISA). The investigation of ISA has up to now been hampered by the lack of a functional marker for ECs in Atlantic salmon in situ. In this study, we report the characterisation and use of a novel monoclonal antibody (MAb) detecting Atlantic salmon ECs (e.g. vessel endothelium, endocardial cells and scavenger ECs) and RBCs. The antibody can be used with immunohistochemistry, IFAT and on Western blots. It appears that the epitope recognised by the antibody is associated with the ISAV cellular receptor. Besides being a tool to identify ECs in situ, it could be useful in further studies of the pathogenicity of ISA. Finally, the detection of an epitope shared by ECs and RBCs agrees with recent findings that these cells share a common origin, thus the MAb can potentially be used to study the ontogeny of these cells in Atlantic salmon.


A complementary marriage of perspectives: understanding organizational social context using mixed methods.
Organizational factors impact the delivery of mental health services in community settings. Mixed-methods analytic approaches have been recommended, though little research within implementation science has explicitly compared inductive and deductive perspectives to understand their relative value in understanding the same constructs. The purpose of our study is to use two different paradigmatic approaches to deepen our understanding of organizational social context. We accomplish this by using a mixed-methods approach in an investigation of organizational social context in community mental health clinics. Nineteen agencies, representing 23 sites, participated. Enrolled participants included 130 therapists, 36 supervisors, and 22 executive administrators. Quantitative data was obtained via the Organizational Social Context (OSC) measure. Qualitative data, comprised of direct observation with spot sampling generated from agency visits, was coded using content analysis and grounded theory. The present study examined elements of organizational social context that would have been missed if only quantitative data had been obtained and utilized mixed methods to investigate if stratifying observations based on quantitative ratings from the OSC resulted in the emergence of differential themes. Four of the six OSC constructs were commonly observed in field observations (i.e., proficiency, rigidity, functionality, stress), while the remaining two constructs were not frequently observed (i.e., resistance, engagement). Constructs emerged related to organizational social context that may have been missed if only quantitative measurement was employed, including those around the physical environment, commentary about evidence-based practice initiatives, leadership, cultural diversity, distrust, and affect. Stratifying agencies by "best," "average," and "worst" organizational social context impacted interpretation for three constructs (affect, stress, and leadership). Results support the additive value of integrating inductive and deductive perspectives in implementation science research. This synthesis of approaches facilitated a more comprehensive understanding and interpretation of the findings than would have been possible if either methodology had been employed in isolation.


A Case Study Evaluating the Efficacy of an Ad Hoc Hospital Collection Device for Fentanyl in Infant Oral Fluid.
Neonatal drug exposure is currently assessed using meconium, urine, blood, hair, or umbilical cord tissue/blood. Due to the invasiveness, challenges, and limitations of collection, and/or analytical difficulties of these matrices, oral fluid may be a more desirable matrix in diagnosing opioid exposure and risk for opioid withdrawal in neonatal abstinence syndrome. Traditional oral fluid collection devices are not viable options as they are too large for neonates' mouths and may contain chemicals on the collection pad. Unstimulated and stimulated infant oral fluid samples have been used for therapeutic drug monitoring as an alternative matrix to blood. The objective of this study was to assess the viability of a simple oral fluid collection system using a sterile foam-tipped swab rinsed in phosphate-buffered saline. Two infants were administered fentanyl for post-operative pain relief while hospitalized in the Neonatal Intensive Care Units at the Children's Hospital of Richmond of Virginia Commonwealth University. Oral fluid samples were collected at 16 h, 2 days, and/or 7 days following the start of intravenous infusion of fentanyl. Samples were analyzed by ultra-high-pressure liquid chromatography-tandem mass spectrometry for fentanyl and norfentanyl after solid-phase extraction. In one of the three samples tested, fentanyl and norfentanyl were detected at concentrations of 28 and 78 ng/mL, respectively. Based on the infusion rate, the theoretical oral fluid fentanyl concentration at steady state was calculated to be 33 ng/mL.


Nitrogen footprint in China: food, energy, and nonfood goods.
The nitrogen (N) footprint is a novel approach to quantify losses to the environment of reactive N (Nr; all species of N except N2) derived from human activities. However, current N footprint models are difficult to apply to new countries due to the large data requirement, and sources of Nr included in calculating the N footprint are often incomplete. In this study, we comprehensively quantified the N footprint in China with an N mass balance approach. Results show that the per capita N footprint in China increased 68% between 1980 and 2008, from 19 to 32 kg N yr(-1). The Nr loss from the production and consumption of food was the largest component of the N footprint (70%) while energy and nonfood products made up the remainder in approximately equal portion in 2008. In contrast, in 1980, the food-related N footprint accounted for 86% of the overall N footprint, followed by nonfood products (8%) and energy (6%). The findings and methods of this study are generally comparable to that of the consumer-based analysis of the N-Calculator. This work provides policy makers quantitative information about the sources of China's N footprint and demonstrates the significant challenges in reducing Nr loss to the environment.


JAK1 and Tyk2 activation by the homologous polycythemia vera JAK2 V617F mutation: cross-talk with IGF1 receptor.
The majority of polycythemia vera (PV) patients harbor a unique somatic mutation (V617F) in the pseudokinase domain of JAK2, which leads to constitutive signaling. Here we show that the homologous mutations in JAK1 (V658F) and in Tyk2 (V678F) lead to constitutive activation of these kinases. Their expression induces autonomous growth of cytokine-dependent cells and constitutive activation of STAT5, STAT3, mitogen-activated protein kinase, and Akt signaling in Ba/F3 cells. The mutant JAKs exhibit constitutive signaling also when expressed in fibrosarcoma cells deficient in JAK proteins. Expression of the JAK2 V617F mutant renders Ba/F3 cells hypersensitive to insulin-like growth factor 1 (IGF1), which is a hallmark of PV erythroid progenitors. Upon selection of Ba/F3 cells for autonomous growth induced by the JAK2 V617F mutant, cells respond to IGF1 by activating STAT5, STAT3, Erk1/2, and Akt on top of the constitutive activation characteristic of autonomous cells. The synergic effect on proliferation and STAT activation appears specific to the JAK2 V617F mutant. Our results show that the homologous V617F mutation induces activation of JAK1 and Tyk2, suggesting a common mechanism of activation for the JAK1, JAK2, and Tyk2 mutants. JAK3 is not activated by the homologous mutation M592F, despite the presence of the conserved GVC preceding sequence. We suggest that mutations in the JAK1 and Tyk2 genes may be identified as initial molecular defects in human cancers and autoimmune diseases.


Thomson scattering diagnostic system design for the Compact Toroidal Hybrid experiment.
A new Thomson scattering system using standard commercially available components has been designed for the non-axisymmetric plasmas of the Compact Toroidal Hybrid (CTH). The beam, generated by a frequency doubled Continuum PL DLS 2 J Nd:YAG laser, is passed vertically through an entrance Brewster window and an aperturing baffle system to minimize the stray laser light that could enter the collection optics. The beam line has been designed with an 8 m propagation distance to the mid-plane of the CTH device with the beam diameter kept less than 3 mm inside the plasma volume. The beam exits the vacuum system through another Brewster window and enters a beam dump, again to minimize the stray light in the vacuum chamber. Light collection, spectral processing, and signal detection are accomplished with an f/#∼ 1 aspheric lens, a commercially available Holospec f/1.8 spectrometer, and an Andor iStar DH740-18U-C3 image intensified camera. Spectral rejection of stray laser light, if needed, can be performed with the use of an optional interference filter at the spectrometer input. The system has been developed for initial single point measurements of plasmas with core electron temperatures of approximately 20-300 eV and densities of 5 × 10(18) to 5 × 10(19) m(-3) dependent upon operational scenario.


Patient perspectives on how to improve education on medication side effects: cross-sectional observational study at a rheumatology clinic in The Netherlands.
Rheumatoid arthritis (RA) patients often report lacking information on medication side effects. The aims of this study were to observe how rheumatology healthcare providers deliver medication information and to determine in which specific domains information is missing. First, 12 single-blinded structured observations were performed during regular RA patient consultations. The observers noted whether and how medication and medication side effects were discussed. Second, 100 RA patients were asked to fill out an adaptation of the Satisfaction with Information about Medicines Scale (SIMS). Medication was discussed during all observed consultations. With new medication, its purpose and mode of action were explained in all cases, but possible side effects in only 33%. Overall, medication side effects were discussed in 58% of consultations. Most information delivery was verbal (92%). Response rate to the questionnaire was 61%. Overall satisfaction with medication education was mean 7.3 (± 1.9) (NRS 0-10) with a comparable high SIMS total satisfaction sum score of mean 12.3 (± 4.4). At subscale score levels, 89% were satisfied with the amount of information on the action and usage of medication, but only 47% with the information on the potential problems of medication. RA patients express overall high satisfaction with their medication education but there is still an unmet need for information on potential risks and side effects. Using the SIMS questionnaire in daily clinical practice may help focus medication education to the needs of the individual patient.


Effect of microbial transglutaminase on spaghetti quality.
To examine the potential application of microbial transglutaminase (MTG) on semolina dough properties and quality of raw and cooked spaghetti, the effects of various MTG addition levels on the solubility of proteins, SDS-PAGE pattern of semolina dough proteins, and textural and structural properties of raw and cooked spaghetti were investigated using semolina from a high-protein good variety (MACS 1967) and a low-protein poor variety (PDW 274) durum wheat. To increase the concentration of lysyl residues and possibly enhance the extent of cross-linking of protein matrix by MTG, a commercial soy protein isolate (SPI) was added at a level of 3% (w/w) in combination with MTG, and its effect on semolina dough properties and spaghetti quality was investigated. The addition of MTG significantly decreased the solubility of semolina dough proteins. SDS-PAGE results showed that with increasing levels of MTG, a progressive decrease in the intensity of the bands corresponding to molecular weight of around 66 kDa was observed. Protein cross-linking reaction catalyzed by MTG resulted in changes in dough properties, dry spaghetti quality, cooking quality characteristics, and microstructure of cooked spaghetti. However, the quality improvements were more evident in spaghetti from the poor variety PDW 274 than from the good variety MACS 1967. The results also showed the ability of MTG in the formation of heterologous polymers between SPI and durum wheat proteins to improve the quality of spaghetti samples.


Cytologic and histologic features of COVID-19 post-vaccination lymphadenopathy.
In this study, we present six cases of axillary or supraclavicular lymphadenopathies in the days following to the Pfizer-BioNTech vaccine's first dose and its study for fine-needle aspiration cytology (two patients), biopsy with a gross needle (two patients), and the evolution of two other patients. Cytological smears and histological sections reveal reactive nodes, predominantly T, CD8, and the presence of atypia with striking mitosis. Differential diagnosis with viral lymphadenopathies and lymphoma is important, so the clinical correlation, in a vaccination context, sudden appearance, and progressive decrease in days of the adenopathy, is essential. In this article, we have studied the cytohistological manifestations of post-Pfizer-BioNTech vaccination lymphadenopathy and discussed its differential diagnosis.


Functional analysis of the novel mycorrhiza-specific phosphate transporter AsPT1 and PHT1 family from Astragalus sinicus during the arbuscular mycorrhizal symbiosis.
Arbuscular mycorrhizas contribute significantly to inorganic phosphate (Pi) uptake in plants. Gene networks involved in the regulation and function of the Pht1 family transporters in legume species during AM symbiosis are not fully understood. In order to characterize the six distinct members of Pht1 transporters in mycorrhizal Astragalus sinicus, we combined cellular localization, heterologous functional expression in yeast with expression/subcellular localization studies and reverse genetics approaches in planta. Pht1;1 and Pht1;4 silenced lines were generated to uncover the role of the newly discovered dependence of the AM symbiosis on another phosphate transporter AsPT1 besides AsPT4. These Pht1 transporters are triggered in Pi-starved mycorrhizal roots. AsPT1 and AsPT4 were localized in arbuscule-containing cells of the cortex. The analysis of promoter sequences revealed conserved motifs in both AsPT1 and AsPT4. AsPT1 overexpression showed higher mycorrhization levels than controls for parameters analysed, including abundance of arbuscules. By contrast, knockdown of AsPT1 by RNA interference led to degenerating or dead arbuscule phenotypes identical to that of AsPT4 silencing lines. AsPT4 but not AsPT1 is required for symbiotic Pi uptake. These results suggest that both, AsPT1 and AsPT4, are required for the AM symbiosis, most importantly, AsPT1 may serve as a novel symbiotic transporter for AM development.


Interfacial Donor-Acceptor Engineering of Nanofiber Materials To Achieve Photoconductivity and Applications.
Self-assembly of π-conjugate molecules often leads to formation of well-defined nanofibril structures dominated by the columnar π-π stacking between the molecular planes. These nanofibril materials have drawn increasing interest in the research frontiers of nanomaterials and nanotechnology, as the nanofibers demonstrate one-dimensionally enhanced exciton and charge diffusion along the long axis, and present great potential for varying optoelectronic applications, such as sensors, optics, photovoltaics, and photocatalysis. However, poor electrical conductivity remains a technical drawback for these nanomaterials. To address this problem, we have developed a series of nanofiber structures modified with different donor-acceptor (D-A) interfaces that are tunable for maximizing the photoinduced charge separation, thus leading to increase in the electrical conductivity. The D-A interface can be constructed with covalent linker or noncovalent interaction (e.g., hydrophobic interdigitation between alkyl chains). The noncovalent method is generally more flexible for molecular design and solution processing, making it more adaptable to be applied to other fibril nanomaterials such as carbon nanotubes. In this Account, we will discuss our recent discoveries in these research fields, aiming to provide deep insight into the enabling photoconductivity of nanofibril materials, and the dependence on interface structure. The photoconductivity generated with the nanofibril material is proportional to the charge carriers density, which in turn is determined by the kinetics balance of the three competitive charge transfer processes: (1) the photoinduced electron transfer from D to A (also referred to as exciton dissociation), generating majority charge carrier located in the nanofiber; (2) the back electron transfer; and (3) the charge delocalization along the nanofiber mediated by the π-π stacking interaction. The relative rates of these charge transfer processes can be tuned by the molecular structure and nanoscale interface engineering. As a result, maximal photoconductivity can be achieved for different D-A nanofibril composites. The photoconductive nanomaterials thus obtained demonstrate unique features and functions when employed in photochemiresistor sensors, photovoltaics and photocatalysis, all taking advantages of the large, open interface of nanofibril structure. Upon deposition onto a substrate, the intertwined nanofibers form networks with porosity in nanometer scale. The porous structure enables three-dimensional diffusion of molecules (analytes in sensor or reactants in catalysis), facilitating the interfacial chemical interactions. For carbon nanotubes, the completely exposed π-conjugation facilitates the surface modification through π-π stacking in conjunction with D-A interaction. Depending on the electronic energy levels of D and A parts, appropriate band alignment can be achieved, thus producing an electric field across the interface. Presence of such an electric field enhances the charge separation, which may lead to design of new type of photovoltaic system using carbon nanotube composite.


Identification and genome analysis of tomato chlorotic spot virus and dsRNA viruses from coinfected vegetables in the Dominican Republic by high-throughput sequencing.
The Tomato chlorotic spot virus (TCSV) was first reported in the 1980s, having its occurrence limited to Brazil and Argentina. Due to an apparent mild severity in the past, molecular studies concerning TCSV were neglected. However, TCSV has disseminated over the USA and Caribbean countries. In Dominican Republic TCSV has been recently reported on important cultivated crops such as pepper and beans. In this work, we provide the first complete genome of a TCSV isolate from symptomatic plants in Dominican Republic, which was obtained by high-throughput sequencing. In addition, three dsRNA viruses from different virus families were identified coinfecting these plants Bell pepper endornavirus (BPEV), Southern tomato virus (STV) and Pepper cryptic virus 2 (PCV-2). Phylogenetic analysis showed that the Dominican Republic TCSV isolate has a close relationship with other TCSV isolates and a reassortant isolate between TCSV and Groundnut ringspot virus (GRSV), all found in USA. BPEV, STV and PCV-2 isolates from Dominican Republic were close related to corresponding American isolates. The possible biological implications of these virus-mixed infections are discussed.