Temporal and spatial characteristics of evolving cell injury during regional myocardial ischemia in the dog: the "border zone" controversy.
An open chest dog heart with multiple coronary ligations was used to define the temporal and spatial characteristics of injury evolving during regional ischemia. With the use of a multiple (40 sample) biopsy device, adjacent transmural biopsy specimens were obtained from the transition zone between normal and ischemic tissue after 5, 30, 45, 60 and 120 minutes of ischemia. The first 1.8 mm of epicardial tissue was taken for the analysis of flow and metabolites. The results confirmed the existence of a sharp interface of flow and metabolism in the epicardial lateral plane at the boundary of the ischemic zone. There was no significant zone of intermediate injury (flow and metabolism being depressed uniformly throughout the ischemic area). Comparison of the distribution of flow determined by radiolabeled gadolinium-153 at onset of ischemia with that indicated by radiolabeled tin-113 microspheres given at the end of various periods of ischemia revealed no change in the position or steepness of the flow interface at any time during the first 2 hours of ischemia. This observation, together with the absence of any major redistribution or enhancement of residual flow to the ischemic zone, indicated that there was little or no significant collateralization between 5 and 120 minutes. Analysis of the adenosine triphosphate (ATP) content revealed a rapid depletion during the first 5 minutes of ischemia; the content then remained essentially unchanged until 30 minutes, after which time a second phase of accelerated ATP depletion was observed until 45 minutes. ATP content then remained relatively constant up to 2 hours.


Prevalence of gastro-oesophageal reflux disease, and its associated risk factors among medical students: a nation-based cross-sectional study.
Gastroesophageal reflux disease (GERD) is a common digestive problem in adults particularly medical students, who are one of the most vulnerable groups. Many variables, including lifestyle changes and psychological stress, increase the prevalence of GERD among undergraduate medical students. Therefore, this study aims to assess the prevalence, and risk factors of GERD, and its relationship with perceived stress among medical students in Egypt. In November and December of 2022, a cross-sectional descriptive study with an analytical component was carried out among medical students from six different universities. An online self-reported questionnaire was used to collect data. The questionnaire included data on sociodemographic characteristics, risk factors, lifestyle, the Arabic version of GerdQ, and the Arabic version of Cohen's Perceived Stress Scale (PSS). The questionnaire was filled out by 964 medical students, the majority of whom were female (64%). Overall, 17.1% of participants reported symptoms of GERD. Logistic regression showed that smoking, high perceived stress, and a family history of GERD were the independent predictors of having GERD symptoms, with odds ratios of 4.1, 3.9, and 2.2, respectively. GERD is a frequent condition among Egyptian medical students, affecting around one-fifth of them. In the fight against GERD, university-based smoking cessation and stress management programs may be influential.


Genetic and epigenetic alterations in MGMT gene and correlation with concomitant chemoradiotherapy (CRT) in cervical cancer.
The MGMT (O6-methylguanine-DNA methyltransferase) gene plays a crucial role in repairing DNA damage caused by alkylating agents, including those used in chemotherapy. Genetic and epigenetic alterations can influence the regulation of MGMT gene, which in turn may impact the response to concomitant chemoradiotherapy (CRT) in cervical cancer. The present study was undertaken to evaluate the correlation of such variations in MGMT gene with the treatment outcome of concomitant chemoradiotherapy (CRT) in cervical cancer. A total of 460 study subjects (240 controls and 220 patients) were subjected to genotypic analysis of MGMT gene variants rs12917(T/C) and rs2308327(A/G) by Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR). Out of them, 48 each of controls and patients were analyzed for promoter methylation and expression by methylation-specific PCR and real-time PCR, respectively. Patients (n = 48) were followed up and evaluated for treatment (CRT) outcome. Statistical analyses were done using GraphPad (9.0) and SPSS version 18.0. Individuals with GG genotype, G allele of rs2308327, and haplotype 'TA' of both variants showed a significant increase in the development of cervical cancer (P ≤ 0.05). In epigenetic regulation, there was a significant hypermethylation of MGMT gene and down-regulation of their expression in patients compared to control individuals. In treatment outcome of CRT, GG genotype of rs2308327(A/G) gene variant showed better response and GG + AG was significantly associated with vital status (alive). Unmethylated MGMT gene showed better median overall survival up to 25 months significant in comparison to methylated MGMT promoter. Gene variant rs2308327(A/G) and promoter hypermethylation regulated MGMT gene can be a good prognostic for treatment response in cervical cancer patients.


Influence of mono- and multivalent cations on the electrokinetic properties of Rana pipiens kidney cells.
Changes in the cation composition of the ambient cell environment, at constant ionic strengths, altered the electrokinetic properties of the surface of normal adult Rana pipiens kidney cells in a manner which reflected its valence. the effect was expressed in the changes of the magnitude of the net surface charge, at any pH, and in the shift of the isoelectric point of the cell surface.


Bactericidal Pectin/Chitosan/Glycerol Films for Food Pack Coatings: A Critical Viewpoint.
Pectin and chitosan films containing glycerol (Gly) at 5, 10, 15, 20, 30, and 40 wt % were prepared in an aqueous HCl solution (0.10 M) by the solvent evaporation method. The unwashed film (UF) containing 40 wt % Gly (UF40) had elongation at break (ε, %) of 19%. Washed films (WFs) had high tensile strength (σ > 46 MPa) and low elongation at break (ε, <5.0%), enabling their use in food packaging applications. The polymers' self-assembling occurred during the washing, increasing the stiffness. The XPS analysis suggests that some HCl is lost during the drying process, resulting in a low acid content on the UF surfaces. The UF40 (at 5.0 mg/mL) exhibits cytocompatibility toward mammalian cells and antimicrobial and anti-adhesive properties against Escherichia coli. The remaining HCl in the UF40 can be a disadvantage for food packaging applications; the UF40 (∅ = 8.5 mm; 55 μm thickness) releases H3O+/HCl, reducing the pH to approximately 3.0 when kept in 200 mL distilled water for approximately 30 min. Therefore, we propose the use of UF40 to coat commercial food packaging. The UF40 has low permeability to water vapor and oxygen and works as a barrier against ultraviolet light. The UF40 is also colorless and completely transparent. The UF40 maintained tomatoes' structural integrity for 18 days at room temperature with no oxidation or microorganism contamination. This paper presents a critical viewpoint concerning chitosan-based films with antimicrobial activities.


In-situ preparation of sulfonated carbonaceous copper oxide-zirconia nanocomposite as a novel and recyclable solid acid catalyst for reduction of 4-nitrophenol.
The missing-linker defects of UiO-66 were exploited to covalently anchor Cu nanoclusters (Cu/UiO-66). The molecular interactions between the metals and oxides as copper-zirconia interfaces in Cu/UiO-66 are essential for heterogeneous catalysis, leading to remarkable synergistic impacts on activity and selectivity. Homogeneously distributed carbonaceous mixed metal oxides (CuO/ZrO2@C) nanocomposite was prepared via carbonization of the Cu/UiO-66 at 600 °C for 3 h in air. To enhance the acidity properties of the CuO/ZrO2@C nanocomposite, a small amount of sulfuric acid was added and heated at 150 °C under an N2 atmosphere (CuO/ZrO2-SO3H@C). The synthesised Cu/UiO-66 and CuO/ZrO2-SO3H@C catalysts were used as novel catalysts in the reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP). The Cu/UiO-66 and CuO/ZrO2-SO3H@C catalysts displayed complete conversion of the 4-NP solution during (4 and 2 min) stirring at room temperature, respectively. These two catalysts exhibited a high reduction rate of 8.61 × 10-3 s-1, and 18.3 × 10-3 s-1, respectively. The X-ray photoelectron spectroscopic (XPS) analysis showed the charge of copper atoms in the Cu/UiO-66 catalyst was Cu0/CuII and in the CuO/ZrO2-SO3H@C catalyst was CuI/CuII with nearly the same ratio (65/35). The particle size and the elemental composition of the CuO/ZrO2-SO3H@C catalyst were analysed by using high resolution transmission electron microscopy (HR-TEM), and energy-dispersive X-ray spectroscopy (EDS), and elemental mapping, respectively. The key point beyond the high catalytic activity and selectivity of the CuO/ZrO2-SO3H@C catalyst is both the carbon-metal oxides heterojunction structure that leads to good dispersion of the CuO and ZrO2 over the carbon sheets, and the high acidity properties that come from the combination between the Brønsted acid sites from sulfuric acid and Lewis acid sites from the UiO-66. The catalysts exhibited good recyclability efficiency without significant loss in activity, indicating their good potential for industrial applications.


Inflammation, a link between obesity and atrial fibrillation.
Despite the long belief that the role of the adipose tissue was restricted to that of a passive store of triglycerides and a rich source of fatty acids, accumulating data demonstrates that the adipose tissue acts as an endocrine organ, capable of producing a large number of cytokines incriminated in generating a systemic inflammatory status. At its turn, this adiposity-related pro-inflammatory status appears to promote a large range of cardiovascular disorders, including atrial fibrillation (AF). Recent studies suggest that, in addition to systemic adiposity, the volume of the pericardial fat of the entire heart, and particularly of that overlying the atria, may represent an even more important risk factor for AF. This review focuses on the most relevant clinical and experimental data that bridge adiposity-induced inflammation and AF, and provides, through a multidisciplinary approach, a discussion that integrates both the current knowledge regarding the prolific activity of systemic and pericardial adipose tissue as sources of inflammatory mediators and the main effects of adiposity-induced inflammation on the most relevant electrophysiological, structural, and autonomic mechanisms responsible for AF.


Design and synthesis of potent vitamin D receptor antagonists with A-ring modifications: remarkable effects of 2alpha-methyl introduction on antagonistic activity.
Novel A-ring analogues of the vitamin D receptor (VDR) antagonist (3a), ZK-159222, and its 24-epimer (3b) were convergently synthesized. Preparation of the CD-ring portions with the side chains of 3a,b, followed by palladium-catalyzed cross-coupling with the A-ring enyne precursors (15a,b), (3S,4S,5R)- and (3S,4S,5S)-bis[(tert-butyldimethylsilyl)oxy]-4-methyloct-1-en-7-yne, afforded the 2alpha-methyl-introduced analogues (4a,b) and their 3-epimers (5a,b). The biological profiles of the hybrid analogues were assessed in terms of affinity for VDR, and antagonistic activity to inhibit HL-60 cell differentiation induced by the natural hormone, 1alpha,25-dihydroxyvitamin D(3). The analogue 4a showed an approximately fivefold higher antagonistic activity compared with 3a. The 2alpha-methyl introduction into 3a increased the receptor affinity, thereby enhancing VDR antagonism. This approach to design potent antagonists based on hybridization of structural motifs in the A-ring and in the side chain may prove to be valuable.


Impact of diabetes mellitus on arterial stiffness in a representative sample of an urban Brazilian population.
Independent of other cardiovascular (CV) risk factors, increased arterial stiffness has been established as a predictor of morbidity and mortality. The main aim of this study was to investigate the impact of diabetes on arterial stiffness in a representative sample of an urban Brazilian population plus Amerindians. A total of 1,415 individuals from the general population were randomly selected plus 588 Amerindians from a native community in Brazil. In addition, a sub-sample of 380 individuals from the general population had 5-year follow-up data. Pulse wave velocity (PWV) was measured with a non-invasive automatic device (Complior, Colson; Garges les Gonesses, France) and increased arterial stiffness was defined as PWV ≥ 12 m/s. In the overall group, diabetic individuals had higher frequencies of increased arterial stiffness and hypertension. They also had higher values of PWV, body mass index, total cholesterol, triglycerides, systolic and diastolic blood pressures compared to non-diabetic individuals (p < 0.01). In an analysis stratified by hypertension, PWV values and increased arterial stiffness frequency were higher in diabetic individuals in both groups (hypertensive and non-hypertensive) (p < 0.05). Furthermore, higher risk for increased arterial stiffness was observed in the diabetic individuals from the overall group (OR = 2.27; CI = 1.47-3.52, p < 0.001) and from the hypertensive group (OR = 2.70; CI = 1.58-4.75, p < 0.001), adjusted for covariates. Regarding the ethnic stratification, diabetic individuals from Amerindian, White, and Mulatto (mixed-race) groups had higher PWV values and a greater frequency of increased arterial stiffness compared to non-diabetic individuals. Both diabetic and non-diabetic individuals had higher PWV values after 5 years. There was no significant difference in the 5-year PWV progression in diabetic compared to non-diabetic individuals. These results confirm, in a sample of Brazilian population, that the presence of diabetes is associated with increased arterial stiffness and it may contribute in part to increased cardiovascular risk in diabetic patients.


[Acute carbon monoxide poisoning in the emergency service. The importance of early signs. 90 cases].
The authors describe 90 cases of carbon monoxide poisoning admitted to their unit. The high incidence of accidental poisoning may be partly related to the energy crisis. They underline the importance of recognising the early symptoms, headache, vomiting, weakness of the legs, allowing early diagnosis before irreversible complication set in.


[Variations of heterochromatic chromosomal regions and chromosome abnormalities in children with autism: identification of genetic markers in autistic spectrum disorders].
In the present study, the cytogenetic and molecular cytogenetic analysis of 90 children with autism and their mothers (18 subjects) was carried out. Chromosome fragility and abnormalities were found in four cases: mos 47,XXX[98]/ 46,XX[2]; 46,XY,r(22)(p11q13); 46,XY,inv(2)(p11.2q13),16qh-; 46Y,fra(X)(q27.3)16qh-. Using C-banding and quantitative fluorescent in situ hybridization (FISH), the significantly increased incidence of heterochromatic region variation was shown in autism as compared to the controls (48 and 16%, respectively). Pericentric 9phqh inversion was not characteristic of the patients with autism whereas heterochromatic variations 1phqh, 9qh+ and 16qh- were more frequent in autism (p<0,05). Basing on the data obtained, a possible role of position effect in autism pathogenesis as well as a potential of heterochromatic region variation analysis for the search of biological markers of autistic spectrum disorders are discussed.


Presence of nerve cell bodies in the lingual nerve in the third molar area.
The microanatomy of the lingual nerve in the third molar area was studied. Twenty-two pairs of lingual nerves were dissected from human cadavers. The area of each nerve adjacent to the third molar was identified, cut, serially sectioned, and stained with hematoxylin and eosin. Nerve cell bodies were found within the structure of 40 of the 44 individual nerves (90.91%). There were two patterns of organization of the nerve cell bodies: isolated nerve cell bodies and ganglion-like clusters of nerve cell bodies. Because of the proximity of the submandibular ganglion, it can be inferred that these cell bodies are parasympathetic in function. The presence of nerve cell bodies in the lingual nerve in the third molar region could have possible clinical ramifications.


Purification and characterization of a novel mannitol dehydrogenase from a newly isolated strain of Candida magnoliae.
Mannitol biosynthesis in Candida magnoliae HH-01 (KCCM-10252), a yeast strain that is currently used for the industrial production of mannitol, is catalyzed by mannitol dehydrogenase (MDH) (EC 220.127.116.11). In this study, NAD(P)H-dependent MDH was purified to homogeneity from C. magnoliae HH-01 by ion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography. The relative molecular masses of C. magnoliae MDH, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography, were 35 and 142 kDa, respectively, indicating that the enzyme is a tetramer. This enzyme catalyzed both fructose reduction and mannitol oxidation. The pH and temperature optima for fructose reduction and mannitol oxidation were 7.5 and 37 degrees C and 10.0 and 40 degrees C, respectively. C. magnoliae MDH showed high substrate specificity and high catalytic efficiency (k(cat) = 823 s(-1), K(m) = 28.0 mM, and k(cat)/K(m) = 29.4 mM(-1) s(-1)) for fructose, which may explain the high mannitol production observed in this strain. Initial velocity and product inhibition studies suggest that the reaction proceeds via a sequential ordered Bi Bi mechanism, and C. magnoliae MDH is specific for transferring the 4-pro-S hydrogen of NADPH, which is typical of a short-chain dehydrogenase reductase (SDR). The internal amino acid sequences of C. magnoliae MDH showed a significant homology with SDRs from various sources, indicating that the C. magnoliae MDH is an NAD(P)H-dependent tetrameric SDR. Although MDHs have been purified and characterized from several other sources, C. magnoliae MDH is distinguished from other MDHs by its high substrate specificity and catalytic efficiency for fructose only, which makes C. magnoliae MDH the ideal choice for industrial applications, including enzymatic synthesis of mannitol and salt-tolerant plants.


Reliability of motion measurements after total disc replacement: the spike and the fin method.
As motion preservation is one of the main postulated advantages after total disc replacement (TDR) of the lumbar spine, the quantification of the mobility after TDR seems of special clinical interest. Yet, the best method to assess range of motion (ROM) after TDR remains unclear. The aim of the study was the calculation of 95%-confidence intervals (95%-C.I.) for the measurement error accompanying: (1) different methods (2) different observers and (3) different levels of training for radiographic motion analysis after TDR. In 12 patients the level L4-L5 and in another 12 patients level L5-S1 were measured with the Cobb and the superimposition method on flexion-extension X-rays after monosegmental TDR. Both methods were adopted as the landmarks used the spikes of the prosthesis instead the endplates (spike method) and the fin of the prosthesis instead the whole vertebral body (fin method). Measurements were performed by two experienced (O-I and O-III) and one inexperienced observer (O-II). The adopted spike and fin method showed a better reliability compared to the reported results of the original Cobb and superimposition method. The method used was not clinically relevant for the intraobserver reliability in the experienced observer (95%-C.I.: +/-2.0 degrees for the fin and +/-2.1 for the spike method) and for the interobserver reliability for two experienced observers (95%-C.I.: -2.8 degrees /+2.8 degrees for the fin and -2.9 degrees /+3.1 degrees for the spike method). The intraobserver reliability for the inexperienced observer was inferior for both methods compared to the experienced observer but no clinically relevant differences could be observed in interobserver reliability measures. The spike and fin method are reliable methods for study protocols dealing with angular motion after TDR as clinically valid conclusions can be drawn with an accuracy of about +/-2 degrees for the same observer and with an accuracy of about +/-3 degrees for a different observer.


17-AAG, an Hsp90 inhibitor, ameliorates polyglutamine-mediated motor neuron degeneration.
Heat-shock protein 90 (Hsp90) functions as part of a multichaperone complex that folds, activates and assembles its client proteins. Androgen receptor (AR), a pathogenic gene product in spinal and bulbar muscular atrophy (SBMA), is one of the Hsp90 client proteins. We examined the therapeutic effects of 17-allylamino-17-demethoxygeldanamycin (17-AAG), a potent Hsp90 inhibitor, and its ability to degrade polyglutamine-expanded mutant AR. Administration of 17-AAG markedly ameliorated motor impairments in the SBMA transgenic mouse model without detectable toxicity, by reducing amounts of monomeric and aggregated mutant AR. The mutant AR showed a higher affinity for Hsp90-p23 and preferentially formed an Hsp90 chaperone complex as compared to wild-type AR; mutant AR was preferentially degraded in the presence of 17-AAG in both cells and transgenic mice as compared to wild-type AR. 17-AAG also mildly induced Hsp70 and Hsp40. 17-AAG would thus provide a new therapeutic approach to SBMA and probably to other related neurodegenerative diseases.


Optimization of hydrolysis conditions of Palmaria palmata to enhance R-phycoerythrin extraction.
In this study, response surface methodology was applied to optimize R-phycoerythrin extraction from the red seaweed Palmaria palmata, using enzymatic digestion. Several algal treatments prior to digestion were first investigated. The extraction yield and the purity index of R-phycoerythrin, and the recovery of proteins and reducing sugars in the water-soluble fraction were then studied in relation to the hydrolysis time, the temperature and the enzyme/seaweed ratio. Enzymatic digestion appears to be an effective treatment for R-phycoerythrin extraction. Moreover, using the seaweed roughly cut in its wet form gives the most interesting results in terms of extract quality and economic cost. The R-phycoerythrin extraction yield is 62 times greater than without enzyme treatment and 16 times greater than without optimization. Enzymatic optimization enhanced the purity index up to 16 times.


Alkenes as azido precursors for the one-pot synthesis of 1,2,3-triazoles catalyzed by copper nanoparticles on activated carbon.
A one-pot protocol for the synthesis of 1,2,3-triazoles has been developed starting from inactivated alkenes and based on two click reactions: the azidosulfenylation of the carbon-carbon double bond and the copper-catalyzed azide-alkyne cycloaddition (CuAAC). High yields of the β-methylsulfanyl triazoles have been attained using CuNPs/C as catalyst, with other commercial copper catalysts being completely inactive. The versatility of the methylsulfanyl group has been demonstrated through a series of synthetic transformations, including direct access to 1-vinyl and 4-monosubstituted triazoles.


Constitutive calcium-independent release of Toxoplasma gondii dense granules occurs through the NSF/SNAP/SNARE/Rab machinery.
The signals and the molecular machinery mediating release of dense matrix granules from pathogenic protozoan parasites are unknown. We compared the secretion of the endogenous dense granule marker GRA3 in Toxoplasma gondii with the release of a stably transfected foreign reporter, beta-lactamase, that localizes to parasite dense granules. Both proteins were released constitutively in a calcium-independent fashion, as shown using both intact and streptolysin O-permeabilized parasites. N-Ethylmaleimide and recombinant bovine Rab-guanine dissociation inhibitor inhibited beta-lactamase secretion in permeabilized parasites, whereas recombinant hamster N-ethylmaleimide-sensitive fusion protein and bovine alpha-SNAP augmented release. Guanosine 5'-3-O-(thio)triphosphate, but not cAMP, augmented secretion in the presence but not in the absence of ATP. The T. gondii NSF/SNAP/SNARE/Rab machinery participates in dense granule release using parasite protein components that can interact functionally with their mammalian homologues.


Mothers' experience of having a child with cerebral palsy. A systematic review.
To explore, synthesise and present findings of qualitative studies describing the experiences of mothers raising a child with cerebral palsy. A systematic literature review of the qualitative evidence. A systematic search for qualitative studies published in the following databases: CINAHL (EBSCO), Medline via OVID, SCOPUS, and Google Scholar. The authors independently assessed eligibility, appraised methodological quality using the Critical Appraisal Skills Program tool for qualitative Research (CASP). An inductive thematic analysis method was adopted to synthesise major findings and to construct core concepts and themes. Five overarching themes reflecting the experiences and perceptions of mothers raising and caring for a child with cerebral palsy are developed: 1) adapting and making sacrifices; 2) guilt and cultural blame; 3) social stigma and marginalisation; 4) physical, environmental, and financial challenges and 5) healthcare experiences.


Evaluation of Gradient Concentration Strips for Detection of Terbinafine Resistance in Trichophyton spp.
The number of dermatophytosis cases resistant to terbinafine is increasing all over the world. Therefore, there is a need for antifungal susceptibility testing of dermatophytes for better management of the patients. In the present study, we have evaluated a gradient test (GT) method for testing the susceptibility of dermatophytes to terbinafine. MIC values to terbinafine determined by the EUCAST reference technique and by gradient test were compared for 79 Trichophyton spp. isolates. Overall, MICs were lower with gradient test (MIC50 of 0.002 μg/mL) than with EUCAST (MIC50 of 0.016 μg/mL). Good categorical agreement (>90%) between the 2 techniques was obtained but the essential agreement was variable depending on the batch of gradient test.


Preparation of Pd Nanoparticles Stabilized by Modified Montmorillonite for Efficient Hydrodeoxygenation of Lignin-Derived Phenolic Compounds in Water.
Developing a new and efficient catalytic route for the production of alkanes by upgrading the aqueous phenolic biofuels still remains a challenge. Here, we designed and synthesized a bifunctional catalyst that uses natural montmorillonite (MMT) as support and combines metal active sites and Brӧnsted acid sites in the MMT via ion exchange and reduction roasting process. The catalytic activity of the as-synthesized Pd-MMT (H+) was evaluated by the hydrodeoxygenation (HDO) of a series of lignin-derived phenolic compounds in water. Our model reaction study reveals that the HDO of phenol undergoes an initial hydrogenation of aromatic rings to produce cyclohexanol and cyclohexanone, followed by the dehydration of cyclohexanol to provide intermediate cyclohexene and a final hydrogenation of cyclohexene to create a cyclohexane product. The combination of high metal catalytic activity and Brӧnsted acidity in Pd-MMT (H+) synergistically accelerated the HDO of phenol. Furthermore, good catalytic activity and recycling ability were also observed for other lignin-derived phenolic compounds.


Magnetic graphene dispersive solid phase extraction-ultra performance liquid chromatography tandem mass spectrometry for determination of β-agonists in urine.
In this study, a magnetic graphene-based dispersive solid phase extraction method was first developed for extraction of β-agonists in urine. During the experiments, the absorbent amount, sample pH, extraction time, elution solution and elution time were optimized respectively. The optimized extraction method was finished within 10min, and showed high enrichment factors for 9 β-agonists (20-26 folds). Furthermore, this absorbent could be reused for at least 60 times. Then this extraction method was combined with ultra performance liquid chromatography triple quadrupole tandem mass spectrometry to determine the 9 drugs in urine. The limits of detection for the 9 drugs were in a range of 0.015-0.023ngmL-1, and the recoveries from the standards fortified blank urine were in a range of 60.2%-109.4%. Therefore, this method could be used as a simple, rapid, sensitive and accurate tool to determine trace level of β-agonists in urine.


Percutaneous biopsy of primary tumor in metastatic renal cell carcinoma to predict high risk pathological features: comparison with nephrectomy assessment.
As treatment options evolve for metastatic renal cell carcinoma, there is a need for predictive information to help guide therapy. We assessed the accuracy of percutaneous primary tumor biopsy for metastatic renal cell carcinoma by comparing biopsy findings to final nephrectomy pathology in patients undergoing cytoreductive nephrectomy. Using an institutional database we reviewed the records of patients who underwent percutaneous primary tumor biopsy before cytoreductive nephrectomy. In patients who underwent biopsy elsewhere pathology findings were re-reviewed at our institution. Differences in accuracy based on biopsy technique, imaging modality and biopsy period were determined by chi-square analysis. We identified 166 patients who underwent percutaneous biopsy of the primary tumor before cytoreductive nephrectomy between 1991 and 2007, and had data available for review. Median pathological tumor size was 9.1 cm (range 3 to 32). Median time from biopsy to surgery was 46 days (range 6 to 717). Of 104 patients in whom biopsy was assigned a Fuhrman nuclear grade 33 (31.7%) had the same grade in the nephrectomy specimen, including 74 of 109 (67.9%) when considering only high or low grade. Grade change by more than 2 points was seen in 18 of 104 patients (17.3%). Sarcomatoid features were present in 34 of 166 nephrectomy specimens (20.5%) but only 4 (11.8%) were identified preoperatively. In patients with metastatic renal cell carcinoma percutaneous renal biopsy has poor accuracy to assess Fuhrman nuclear grade or sarcomatoid features. Physicians should use caution when using biopsy data to guide therapy.


Female-enriched expression of ERalpha during gonad differentiation of the urodele amphibian Pleurodeles waltl.
In the amphibian Pleurodeles waltl, estradiol treatment of genetically male larvae (ZZ) induces male-to-female sex reversal whereas heat treatment of genetically female larvae (ZW) inhibits estradiol synthesis and leads to female-to-male sex reversal. No data are available on estrogen receptors in this species. In the present study, we have isolated a unique full-length pwERalpha cDNA and its 5'-flanking region whose promoter activity was confirmed by transfection assays. RT-PCR studies performed in adult animals using ERalpha-specific primers, revealed that pwERalpha mRNA was present mainly in reproductive tissues: gonads, fat body and oviduct. PwERalpha transcript was also detected in liver, suggesting its implication in vitellogenesis control as in numerous oviparous species. The level of pwERalpha transcripts was also studied during gonad differentiation by quantitative real-time PCR. At stage 54(30d) pwERalpha expression in gonads of ZW larvae was significantly higher than in ZZ ones. This sex-specific discrimination was confirmed when gonad-mesonephros-interrenal complexes (GMI), taken at the same stage, were subjected to whole mount in situ hybridization. In comparison, the female-enriched expression of P450 aromatase, which was studied as a control of ovary differentiation, was observed earlier (stage 54). In ZW larvae reared at 32 degrees C, a condition leading to sex reversal, pwERalpha mRNA level at stage 54(30d) was lower than in control females. Taken together, these results showing a female-enriched and thermosensitive expression of pwERalpha suggest an important role for this receptor in gonad differentiation of the urodele amphibian Pleurodeles waltl.


Metaphrase: an aid to the clinical conceptualization and formalization of patient problems in healthcare enterprises.
Patient descriptors, or "problems," such as "brain metastases of melanoma" are an effective way for caregivers to describe patients. But most problems, e.g., "cubital tunnel syndrome" or "ulnar nerve compression," found in problem lists in an Electronic Medical Record (EMR) are not comparable computationally--in general, a computer cannot determine whether they describe the same or a related problem, or whether the user would have preferred "ulnar nerve compression syndrome." Metaphrase is a scalable, middleware component designed to be accessed from problem-manager applications in EMR systems. In response to caregivers' informal descriptors it suggests potentially equivalent, authoritative, and more formally comparable descriptors. Metaphrase contains a clinical subset of the 1997 UMLS Metathesaurus and some 10,000 "problems" from the Mayo Clinic and Harvard Beth Israel Hospital. Word and term completion, spelling correction, and semantic navigation, all combine to ease the burden of problem conceptualization, entry and formalization.


Misoprostol for the termination of pregnancy with a live fetus at 13 to 26 weeks.
A combination of mifepristone and misoprostol is the regimen of choice for termination of pregnancy between 13 to 26 weeks. In many countries, mifepristone is still not available, and misoprostol has to be used alone. Many misoprostol-alone regimens have been reported in the literature with apparently good results. Most of the trials were conducted in pregnancies between 13 and 22 weeks. For this gestational period, we recommend the regimen of 400 microg of vaginal misoprostol every 3 h up to 5 doses, as it appears to be effective without excessive side effects or complications. There is inadequate data to recommend a regimen for the gestational period of 23 to 26 weeks but it is advisable to reduce the dose and frequency of administration of misoprostol. Common side effects of misoprostol-induced termination of pregnancy include gastrointestinal side effects, abdominal cramps, bleeding, fever and chills. Complications may include infection or rarely rupture of uterus.


Effect of a Novel Hybrid Nanocomposite of Cisplatin-Chitosan on Induced Tissue Injury as a Suggested Drug by Reducing Cisplatin Side Effects.
The self-assembly of cisplatin (Cis-Pt) and chitosan nanoparticles (Cs NPs) has been synthesized and characterized successfully by different analyses and techniques, such as scanning electron microscopy, ultraviolet-visible spectrophotometry, and Fourier transform infrared spectroscopy. The efficiency of loading Cis-Pt on Cs NPs for decreasing the side effects of Cis-Pt by loading it on Cs NP surface was revealed through histopathological and physiological measurements for the liver, testis, and kidney cells. Self-assembly hybrid nanocomposite (Cis-Pt@Cs) could improve spermatogenic cells, seminiferous tubules, and Leydig cells in the interstitial tissue. Kidney examination showed intact glomeruli with a mild increase in capsular space in addition to the intact renal tubular epithelial lining, and liver findings showed improvement in dilation and congestion of the central vein besides mild dilation of blood sinusoids in addition to a mild degree of hepatocyte vacuolation. The serum levels of hepatic, renal, and testicular marker analysis were measured, where Cis-Pt increased the serum levels of alanine aminotransferase, aspartate aminotransferase activity, urea, creatinine, and decreased testosterone levels, while synthesized self-assembly appeared normalized levels. From the results, the self-assembly hybrid nanocomposite decreases and improves the side effects of Cis-Pt.


How variations in the composition of urine influence urease-induced crystallization.
To Study how the composition of urine influences urease-induced crystallization, human urine samples were incubated with urease and the subsequent precipitation measured. Beside the pH increase, the urinary content of magnesium and calcium had profound effects on the precipitation of magnesium ammonium phosphate and calcium phosphate, respectively. Urine phosphate, ammonium and osmolarity had no direct effects on the precipitation. Among the urine components with potential inhibitory properties, only albumin was found to be correlated with such an effect. This inhibitory activity was especially influential in urines with high calcium and magnesium levels. These findings suggest that the composition of urine could also influence the formation of stones consisting of magnesium ammonium phosphate and calcium phosphate.


Management of canine perianal fistula with fluorescence light energy: preliminary findings.
Canine perianal fistula (CPF) is a severe, painful, debilitating skin condition affecting the perianal skin. It often interferes with the quality of life of both dog and owner. Conventional medical treatment involves the use of immunosuppressive therapy; however, the successful resolution of lesions can be limited by poor owner compliance, adverse drug effects and dependence on costly therapies. The present study aimed to assess the potential benefits of fluorescent light energy (FLE) on cases of CPF. Four dogs with active perianal fistulas METHODS AND MATERIALS: FLE was applied as sole management therapy once a week with two consecutive applications in the same session for each dog until clinical signs had significantly improved, with weekly assessments for a six week period. Dogs were assessed by measuring the size of lesions at the start of the study and then weekly for six weeks, using planimetry software. Owners recorded vocalization and straining frequency scores during their pet's defaecation, and perianal licking frequency on a 0-5 point scale to assess the response to therapy. All dogs improved with FLE, achieving a significant reduction in vocalization, straining and licking after two weeks (P = 0.002). After five weeks of FLE therapy, lesional areas had significantly decreased (P = 0.04). Only one dog required more than seven applications. No adverse events were reported. FLE may be a promising alternative therapy for CPF.


Genotype-Directed Dosing Leads to Optimized Voriconazole Levels in Pediatric Patients Receiving Hematopoietic Stem Cell Transplantation.
Invasive fungal infections are a significant cause of morbidity and mortality in recipients of hematopoietic stem cell transplantation (HSCT), warranting antifungal prophylaxis as a standard of care in these patients. Voriconazole is commonly used in this setting because of its broad-spectrum activity and available dosage forms. There is wide well-known inter- and intrapatient variability in voriconazole concentrations, in part because concentrations are affected by common CYP2C19 polymorphisms. In 2 successive studies we have optimized voriconazole dosing to achieve target voriconazole serum concentrations using a genotype-specific dosing algorithm for antifungal prophylaxis in the post-HSCT period. In our pilot study all patients undergoing HSCT who received voriconazole antifungal prophylaxis were prospectively followed. Voriconazole concentrations were monitored weekly and doses adjusted until concentrations reached between 1 and 5.5 μg/L. The most common CYP2C19 polymorphisms were determined and correlated with voriconazole dose and time required to reach the target concentration range. In the subsequent study patients receiving voriconazole prophylaxis were dosed based on their CYP2C19 genotype and followed prospectively. In the pilot study 25 patients received voriconazole as antifungal prophylaxis for a median of 49 days (range, 15 to 196 days). The median time to reach the target concentration was 34 days for extensive metabolizers and 11 days for poor metabolizers. Three patients were genotyped as intermediate metabolizers; they reached the target concentration in a median of 56 days. Similarly, 2 patients who were genotyped as ultrarapid metabolizers reached the target range in 18 and 25 days. The time and dose required to reach the adequate concentration showed a trend toward correlation with individual CYP2C19 genotype, although voriconazole concentrations showed large interpatient variability in wild-type patients (extensive metabolizers). In our follow-up study, 20 patients received voriconazole prophylaxis prospectively dosed based on their CYP2C19 genotype. The median times to reach the target concentration using genotype-guided dosing were 9, 6.5, and 4 days for ultrarapid, extensive, and intermediate metabolizers, respectively. Overall, the median time to reach the target concentration with genotype-guided dosing was 6.5 days compared with a median time of 29 days when all patients were started on the same dose regardless of CYP2C19 genotype (P < .001). Our data show that traditional voriconazole dosing does not lead to timely achievement of target levels for fungal prophylaxis. However, a genotype-directed dosing algorithm allows patients to reach the voriconazole target range significantly sooner, providing better prophylaxis against fungal infections in the immediate post-transplant period.


Dexmedetomidine (Dex) exerts protective effects on rat neuronal cells injured by cerebral ischemia/reperfusion via regulating the Sphk1/S1P signaling pathway.
To investigate the influence of dexmedetomidine (Dex) on cerebral ischemia/reperfusion (I/R)-injured rat neuronal cells by regulating the Sphk1/S1P pathway. The rats were divided into the following groups, with 18 rats in each group categorized on the basis of random number tables: sham (Sham), I/R (I/R), Dex, Sphk1 inhibitor (PF-543), and Dex together with the Sphk1 agonist phorbol-12-myristate-13-acetate (Dex+PMA). The neurological functions of the rats were assessed by the Longa scoring system at 24 h post reperfusion. The area of brain infarction was inspected using 2,3,5-triphenyltetrazolium chloride staining, and the water content of brain tissue was determined by the dry-wet weight method. The morphology of neurons in the CA1 region of the rat hippocampus was inspected using Nissl staining, while the apoptosis of neurons in this region was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling staining. The Sphk1 and S1P protein levels were determined by immunofluorescence and western blotting, respectively. Compared to the I/R group, rats in the Dex, PF-543, and Dex+PMA groups had a significantly lower neurological function score, as well as lower brain water content and a decreased infarction area. Moreover, the apoptotic index of the neurons and the Sphk1 and S1P levels in the hippocampal CA1 region were significantly lower in these groups (p<0.05). PMA, an agonist of Sphk1, was able to reverse the protective effects of Dex on I/R-induced neuronal cell injury. Dex could protect cerebral I/R-induced neuronal cell injury by suppressing the Sphk1/S1P signaling pathway.


Molecular cloning, sequence identification and tissue expression profile of three novel genes Sfxn1, Snai2 and Cno from Black-boned sheep (Ovis aries).
The complete coding sequences of three of Black-boned sheep (Ovis aries) genes Sfxn1, Snai2 and Cno were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) according to the conserved sequence information of the cattle or other mammals and known highly homologous sheep ESTs. Black-boned sheep Sfxn1 gene encodes a protein of 322 amino acids which has high homology with the Sfxn1 proteins of five species--cattle 98%, pig 95%, human 95%, rat 93%, and mouse 93%. Black-boned sheep Snai2 gene encodes a protein of 268 amino acids that has high identity with the Snai2 proteins of six species--cattle 99%, pig 94%, human 93%, dog 93%, rat 91%, and mouse 90%. Black-boned sheep Cno gene encodes a protein of 214 amino acids that has high homology with the Cno proteins of four species--cattle 97%, human 75%, mouse 67%, and rat 65%. The phylogenetic tree analysis demonstrated that Black-boned sheep Sfxn1, Snai2 and Cno proteins have close relationship with cattle Sfxn1, Snai2 and Cno proteins. The tissue expression analysis indicated that Black-boned sheep Sfxn1, Snai2 and Cno genes were expressed in a range of tissues including leg muscle, kidney, skin, longissimus dorsi muscle, spleen, heart and liver. Our experiment is the first to provide the primary foundation for further insight into these three sheep genes.


Enhancer of Rudimentary Cooperates with Conserved RNA-Processing Factors to Promote Meiotic mRNA Decay and Facultative Heterochromatin Assembly.
Erh1, the fission yeast homolog of Enhancer of rudimentary, is implicated in meiotic mRNA elimination during vegetative growth, but its function is poorly understood. We show that Erh1 and the RNA-binding protein Mmi1 form a stoichiometric complex, called the Erh1-Mmi1 complex (EMC), to promote meiotic mRNA decay and facultative heterochromatin assembly. To perform these functions, EMC associates with two distinct complexes, Mtl1-Red1 core (MTREC) and CCR4-NOT. Whereas MTREC facilitates assembly of heterochromatin islands coating meiotic genes silenced by the nuclear exosome, CCR4-NOT promotes RNAi-dependent heterochromatin domain (HOOD) formation at EMC-target loci. CCR4-NOT also assembles HOODs at retrotransposons and regulated genes containing cryptic introns. We find that CCR4-NOT facilitates HOOD assembly through its association with the conserved Pir2/ARS2 protein, and also maintains rDNA integrity and silencing by promoting heterochromatin formation. Our results reveal connections among Erh1, CCR4-NOT, Pir2/ARS2, and RNAi, which target heterochromatin to regulate gene expression and protect genome integrity.


A twin study of self-regulatory eating in early childhood: estimates of genetic and environmental influence, and measurement considerations.
Children differ greatly in their ability to self-regulate food intake for reasons that are poorly understood. This laboratory-based twin study tested the genetic and environmental contributions to self-regulatory eating and body fat in early childhood. A total of 69 4-7 year-old same-sex twin pairs, including 40 monozygotic and 29 dizygotic pairs, were studied. Self-regulatory eating was operationalized as the percentage compensation index (COMPX%), assessed by a 'preload' challenge in which lunch intake was measured following a low- (3 kcal) or high-calorie (159 kcal) drink. Body fat indexes also were measured. The familial association for COMPX% was estimated by an intraclass correlation, and biometric analyses estimated heritability. Children ate more at lunch following the low- compared with high-energy preload (P<0.001), although variability in COMPX% was considerable. Compensation was significantly poorer among African American and Hispanic compared with European American children, and among girls compared with boys. There was a familial association for self-regulatory eating (ρ = 0.23, P = 0.03) but no significant genetic component. In all, 22% of the variance in COMPX% was due to shared environmental 'household' factors, with the remaining variance attributable to child-specific 'unique' or 'random' environments. Poorer self-regulatory eating was associated with greater percent body fat (r = -0.21, P = 0.04). Self-regulatory eating was influenced by environmental factors, especially those differing among siblings. The absence of a significant genetic effect may reflect the age of the sample or could be artifactual due to measurement issues that need to be considered in future studies.


Inpatient and outpatient use of the Hickman catheter for adults with osteomyelitis.
One hundred five Hickman catheters were inserted in 96 patients with a diagnosis of osteomyelitis. There were 78 men and 18 women with a mean age of 36.4 years. The catheters were present for a total of 6903 days with a mean of 66.4 days. Sixty-five percent of the catheters were used for outpatient as well as inpatient antibiotic therapy. The mean duration of outpatient antibiotic therapy was 30 days. Nine patients had more than one catheter inserted. The overall complication rate was 20% (21/105 catheters). The infectious complication rate was 11/105 (10.5%) or 0.16/100 catheter days. The noninfectious complication rate was 10/105 (9.5%) or 0.14/100 days. The catheter had to be removed prematurely before the completion of antibiotic therapy in only five of the 105 (4.8%) catheter insertions. The Hickman catheter is a safe and effective intravenous access device for long-term antibiotic therapy in patients with osteomyelitis. Inpatient management and intravenous antibiotic therapy are facilitated by a stable and long-term access to the venous circulation. The simplicity of the device also enables it to be used in the outpatient setting, reducing the time of hospitalization and the costs of treatment.


Inhibitory effects of Ficus carica and Olea europaea on pro-inflammatory cytokines: A review.
Ficus carica (fig) and Olea europaea (olive) are valuable nutritional plants that are widely used in diet and traditional medicine. Different parts of the plants such as fruit and leaves contain beneficial compounds with diverse pharmacological properties, among which anti-inflammatory activities are remarkable. The purpose of this review is to discuss the anti-inflammatory effects of F. carica and O. europaea with emphasis on their impact on pivotal pro-inflammatory cytokines including IL-1, IL-6, and TNF-α. To prepare the present review, the sites utilized included Scopus, PubMed, Science Direct, and Google Scholar and studied relevant articles from 2000 until 2021. As a result, we observed that most of the compounds in fig and olive including polyphenols, flavonoids, etc., exert their anti-inflammatory effects through inhibiting or decreasing pro-inflammatory cytokines. Moreover, some natural antioxidants are common between these two plants. We suggest that consuming figs and olives simultaneously or alone can be useful in the prevention or treatment of inflammatory diseases.


A proposed intergenerational model of substance abuse, family functioning, and abuse/neglect.
Although the link between substance abuse and child maltreatment has been relatively well established, there is a general recognition that this is not a simple cause-effect relationship. The current study explored the relationships among substance abuse, family functioning, and abuse/neglect in a sample of incarcerated substance abusers. Data were gathered on the earlier life experiences of 81 men and women serving sentences in two maximum security prisons, including assessments of their parents' substance abuse problems; levels of family competence within their families-of-origin; their exposure, as children and adults, to physical, emotional, and sexual abuse and physical/emotional neglect; and their own levels of substance abuse. Results showed generally high percentages of parental substance abuse and abuse/neglect, and relatively low levels of family competence. Correlational analyses revealed significant direct and indirect relationships among parental substance abuse, family dynamics, and exposure to both child and adult maltreatment. These four variables were also significantly associated with respondents' own substance abuse in later life, suggesting the potential for continuation of these patterns into successive generations. An intergenerational model of these family and personal functioning variables is presented and implications for service delivery with correctional clients is discussed.


SARS-CoV-2 genomic surveillance in Malaysia: displacement of B.1.617.2 with AY lineages as the dominant Delta variants and the introduction of Omicron during the fourth epidemic wave.
This study reported SARS-CoV-2 whole genome sequencing results from June 2021 to January 2022 from seven genome sequencing centers in Malaysia as part of the national surveillance program. COVID-19 samples that tested positive by reverse transcription polymerase chain reaction and with cycle threshold values <30 were obtained throughout Malaysia. Sequencing of SARS-CoV-2 complete genomes was performed using Illumina, Oxford Nanopore, or Ion Torrent platforms. A total of 6163 SARS-CoV-2 complete genome sequences were generated over the surveillance period. All sequences were submitted to the Global Initiative on Sharing All Influenza Data database. From June 2021 to January 2022, Malaysia experienced the fourth wave of COVID-19 dominated by the Delta variant of concern, including the original B.1.617.2 lineage and descendant AY lineages. The B.1.617.2 lineage was identified as the early dominant circulating strain throughout the country but over time, was displaced by AY.59 and AY.79 lineages in Peninsular (west) Malaysia, and the AY.23 lineage in east Malaysia. In December 2021, pilgrims returning from Saudi Arabia facilitated the introduction and spread of the BA.1 lineage (Omicron variant of concern) in the country. The changing trends of circulating SARS-CoV-2 lineages were identified, with differences observed between west and east Malaysia. This initiative highlighted the importance of leveraging research expertise in the country to facilitate pandemic response and preparedness.


Physiology and morphology of projection neurons in the antennal lobe of the male moth Manduca sexta.
1. We have used intracellular recording and staining, followed by reconstruction from serial sections, to characterize the responses and structure of projection neurons (PNs) that link the antennal lobe (AL) to other regions of the brain of the male sphinx moth Manduca sexta. 2. Dendritic arborizations of the AL PNs were usually restricted either to ordinary glomeruli or to the male-specific macroglomerular complex (MGC) within the AL neuropil. Dendritic fields in the MGC appeared to belong to distinct partitions within the MGC. PNs innervating the ordinary glomeruli had arborizations in a single glomerulus (uniglomerular) or in more than one ordinary glomerulus of one AL (multiglomerular) or in one case, in single glomeruli in both ALs (bilateral-uniglomerular). One PN innervated the MGC and many or all ordinary glomeruli of the AL. 3. PNs with dendritic arborizations in the ordinary glomeruli and PNs associated with the MGC typically projected both to the calyces of the ipsilateral mushroom body and to the lateral protocerebrum, but some differences in the patterns of termination in those regions have been noted for the two classes of PNs. One PN conspicuously lacked branches in the calyces but did project to the lateral protocerebrum. The PN innervating the MGC and many ordinary glomeruli projected to the calyces of the ipsilateral mushroom body and the superior protocerebrum. 4. Crude sex-pheromone extracts excited all neurons with arborizations in the MGC, although some were inhibited by other odors. One P(MGC) was excited by crude sex-pheromone extract and by a mimic of one component of the pheromone blend but was inhibited by another component of the blend. 5. PNs with dendritic arborizations in ordinary glomeruli were excited or inhibited by certain non-pheromonal odors. Some of these PNs also responded to mechanosensory stimulation of the antennae. 6. The PN with dendritic arborizations in the MGC and many ordinary glomeruli was excited by crude sex-pheromone extracts and non-pheromonal odors and also responded to mechanosensory stimulation of the antenna.


Analysis of an agmatine deiminase gene cluster in Streptococcus mutans UA159.
An operon encoding enzymes of the agmatine deiminase system (AgDS) has been identified in the cariogenic bacterium Streptococcus mutans UA159. The AgDS is regulated by agmatine induction and carbohydrate catabolite repression. Ammonia is produced from agmatine at low pH, suggesting that the AgDS could augment acid tolerance.


Penetrating keratoplasty and cataract surgery: the advantages of an extracapsular technique combined with posterior chamber intraocular implantation.
The results of 28 consecutive triple procedures and 33 cases of combined penetrating keratoplasty and extracapsular extraction are presented. All the eyes with the triple procedure maintained a clear graft; 77 per cent achieved an acuity of 6/12 or better. The average time before a refractive correction could be dispensed was 8.2 months, while after a combined procedure the interval was 17.4 months, although in both groups the average time until the best visual acuity was reached was similar.


Application of HPLC-thermospray ionization mass spectrometry for the analysis of triprolidine and its metabolite hydroxymethyltriprolidine in biological samples.
High-performance liquid chromatography coupled to positive ion mode thermospray ionization mass spectrometry has been used to characterize triprolidine and its known metabolite hydroxymethyltriprolidine. The method allows direct analysis of these compounds in complex biological matrices. Analysis of an extract from a microbial biotransformation experiment with triprolidine was performed and the results included two baseline-resolved chromatographic peaks. The [M + H]+ ion obtained from the mass spectrum of the first peak was consistent with that of hydroxymethyltriprolidine, and the mass spectrum of the second peak corresponded to triprolidine. Under the described conditions, the minimum detectable level of triprolidine without filament enhancement was estimated at about 10 micrograms injected on column.


Seizures following picornavirus infection.
We demonstrate the establishment and characterization of a novel virus infection-induced seizure model in C57BL/6 mice. C57BL/6 mice were infected with Theiler's murine encephalomyelitis virus (TMEV) or mock infected. Mice were followed for seizures, weight change, body temperature, motor function (righting reflex, rotorod) and neurological manifestations (inflammation [perivascular cuffing], pyknotic neurons, transforming growth factor [TGF]-beta expression). C57BL/6 mice are susceptible to seizures induced by TMEV infection. Approximately 50% of C57BL/6 mice develop transient afebrile seizures. Motor function and coordination are impaired in seized mice. Pyramidal neuron pyknosis and TGF-beta expression correlate with seizure activity in the hippocampus. The characterization of this model will enable the investigation of viral and immune contributions in the central nervous system to the development of seizure disorders in humans.


IL-4 and IL-13 mRNA real-time PCR quantification on whole blood to assess allergic response.
Although routinely used in clinical practice, skin prick tests and serum specific IgE often fail to distinguish between IgE-sensitization and symptomatic IgE-mediated allergy. There is therefore a need for new laboratory tests relating allergic symptoms to the offending agent. In this way, we evaluated the diagnostic reliability of a new whole blood quantitative real-time PCR assay for IL-4 and IL-13 mRNAs. We compared the response of cat allergic patients and non-cat allergic controls upon anti-IgE and cat allergen (Fel d1) stimulation of whole blood. Allergen addition led to a significant increase of IL-4 and IL-13 mRNAs in allergic patients compared to non-allergic controls (p<0.0001). Both cytokine mRNA levels were strongly correlated and peaked within 2 h after Fel d1 or anti-IgE addition. This rapid increase as well as purification experiments led us to the conclusion that basophils represent an important if not the main source of both transcripts in this setting. The effect was allergen-specific since not observed when stimulating blood from cat allergic patients with birch pollen. Interestingly, we found that IFN-gamma mRNA, contrary to IL-4 mRNA, reached higher levels in response to Fel d1 with control individuals than with patients allergic to cat. This study shows that whole blood real-time PCR is a valuable method for IL-4 and IL-13 mRNA measurement after in vitro allergen challenge. We suggest that it might be useful, complementing conventional markers, for the diagnosis and the follow-up of allergic diseases. Further assessments are required to evaluate the clinical potential of this technique.


Early response predicts a sustained response to eluxadoline in patients with irritable bowel syndrome with diarrhoea in two Phase 3 studies.
The mixed μ- and κ-opioid receptor agonist and δ-opioid receptor antagonist, eluxadoline, is licensed in the USA for the treatment of irritable bowel syndrome with diarrhoea (IBS-D), based on the results of two large Phase 3 clinical trials. To understand the time course of treatment benefits with eluxadoline by comparing responder rates over the first month of treatment with responder rates over longer treatment intervals. In this post hoc analysis of two Phase 3 studies, composite and adequate relief (AR) responder rates were calculated over month 1 and patients were stratified by their responder status. Cumulative counts over subsequent intervals (months 1-3, months 1-6, months 2 through 6) were tallied. The studies randomised 2428 patients. Over month 1, 24.6%, 22.8% and 12.5% of patients were composite responders with eluxadoline 100 mg, eluxadoline 75 mg and placebo respectively. For month 1 responders, 77.8% and 81.5% (over months 1-3) and 70.7% and 73.9% (over months 1-6) showed a continuous response with eluxadoline 100 mg and 75 mg respectively. [Correction added on 5 April 2017, after first online publication: The percentage for the responders over months 1-3 was previously wrong and has been corrected.] Of the month 1 nonresponders, <20% showed a response over months 1-3 or months 1-6. Similar results were seen for the analysis of proportions of AR responders over these time intervals. Over two-thirds of patients who respond over the first month retain a positive response over 6 months of treatment with eluxadoline, indicating that early clinical response to eluxadoline is associated with sustained benefits for up to 6 months in patients with IBS-D.


Role of Pax6 in forebrain regionalization.
Pax6 is a highly conserved transcription factor essential for the development of the eyes in vertebrate and invertebrate species. It is also required for normal development of many regions of the central nervous system, including the mammalian forebrain, hindbrain and spinal cord. In the forebrain, it is expressed in a gradient in the dorsal telencephalon, where it is required for the expression of genes that confer dorsal characteristics and where it might play a role in regionalization of the cerebral cortex. It is expressed in the diencephalon, where it is essential for the specification of its derivatives. While the ancestral function of Pax6 may have been to specify a structure sensitive to light, it has been co-opted into the regulation of a broader range of processes in development of the vertebrate nervous system.


A Randomized Trial of Deep Brain Stimulation to the Subcallosal Cingulate and Nucleus Accumbens in Patients with Treatment-Refractory, Chronic, and Severe Anorexia Nervosa: Initial Results at 6 Months of Follow Up.
The main objective of this study was to assess the safety and efficacy of deep brain stimulation (DBS) in patients with severe anorexia nervosa (AN). Eight participants received active DBS to the subcallosal cingulate (SCC) or nucleus accumbens (NAcc) depending on comorbidities (affective or anxiety disorders, respectively) and type of AN. The primary outcome measure was body mass index (BMI). Overall, we found no significant difference (p = 0.84) between mean preoperative and postoperative (month 6) BMI. A BMI reference value (BMI-RV) was calculated. In patients that received preoperative inpatient care to raise the BMI, the BMI-RV was defined as the mean BMI value in the 12 months prior to surgery. In patients that did not require inpatient care, the BMI-RV was defined as the mean BMI in the 3-month period before surgery. This value was compared to the postoperative BMI (month 6), revealing a significant increase (p = 0.02). After 6 months of DBS, five participants showed an increase of ≥10% in the BMI-RV. Quality of life was improved (p = 0.03). Three cases presented cutaneous complications. DBS may be effective for some patients with severe AN. Cutaneous complications were observed. Longer term data are needed.


Infectious diseases. NH and MRC report. From the 91st session of the National Health and Medical Research Council, June 4 and 5, 1981.
The National Health and Medical Research Council has revised its recommendations regarding periods of exclusion from school of infectious disease cases and contacts; vaccination of preterm infants; childhood immunisation schedules; rubella immunisation; treponemal serology testing; treatment of gonorrhoea.


Further evidence for the location of the blepharophimosis syndrome (BPES) at 3q22.3-q23.
We report a 6-year-old, mentally retarded boy with typical clinical signs and symptoms of the blepharophimosis syndrome (blepharophimosis, ptosis, epicanthus inversus syndrome (BPES)), born to normal parents. Chromosome studies revealed an interstitial deletion in the long arm of chromosome 3: del(3)(q22.3-->q23). This observation reinforces previous suggestions that the location of the BPES gene is at 3q2, i.e. 3q22.3-q23.


Molecular characterization of human and animal Echinococcus granulosus isolates in Isfahan, Iran.
Cystic hydatid disease (CHD) is one of the most important zoonotic diseases in different parts of Iran. While it causes major health problem, there is limited information about its transmission cycles and reservoirs of human infection. Therefore we aimed to characterize the existence Echinococcus granulosus cysts in humans and animals in the province of Isfahan, central region of Iran. We collected hydatid cysts from the liver and lungs of patients who underwent surgery procedure and also cysts were obtained from domestic animals at slaughterhouses. DNA was extracted from the protoscoleces and examined by polymerase chain reaction (PCR) of rDNA internal transcribed spacer1 (ITS1-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). In addition, fragments of the genes coding for mitochondrial cytochrome c oxidase subunit 1 (CO1) and NADH dehydrogenase 1 (ND1) were sequenced. Among the two different identified strains/genotypes (sheep and camel), the sheep strain was shown to be the most common genotype of E. granulosus affecting humans, sheep, cattle, goats and occasionally camels. Nine out of 26 camel samples were infected with sheep strain. However the camel genotype was observed in humans, camels and cattle. Seventeen out of 26 camel isolates, 6 out of 31 human and 5 out of 14 cattle samples were infected with the camel genotype. The camel genotypes had PCR and RFLP patterns which were different from the PCR and RFLP patterns of the rest of isolates (sheep strain). The results of this study showed that the 'camel' strain was actual source of infection to humans which circulates between intermediate hosts including camels and cattle, and it confirms the camel-dog transmission cycle in Isfahan.


Using the maternal-fetal genotype incompatibility test to assess non-inherited maternal HLA-DRB1 antigen coding alleles as rheumatoid arthritis risk factors.
Non-inherited maternal antigens encoded by specific HLA-DRB1 alleles (NIMA) have been implicated as a rheumatoid arthritis (RA) risk factor. Using genotype data from North American Rheumatoid Arthritis Consortium study participants and the maternal-fetal genotype incompatibility (MFG) test, we find evidence for offspring allelic effects but no evidence for NIMA as a RA risk factor. We discuss possible reasons why our result conflicts with several previous studies (including one of our own) that used RA patients from northern Europe.


Assessment of the risk of heat disorders encountered during work in hot conditions.
To co-ordinate the work of the main European research teams in the field of thermal factors in order to develop and improve significantly the methods presently available for assessing the risks of heat disorders encountered during work in hot conditions. Each item from the required sweat rate model was reviewed on the basis of the most recent literature. A database with 1,113 laboratory and field experiments, covering the whole range of hot working conditions, was assembled and used for the validation. Influence of clothing ensemble on heat exchange: methods and formulas were developed that take into account the dynamic effects associated with forced convection and the pumping effect associated with body movements and exercise. Prediction of the average skin temperature: the model used in the required sweat rate standard ISO 7933 was extended to cover more severe conditions with high radiation and high humidity and different clothing and take into account the rectal temperature for the prediction of the skin temperature. Criteria for estimating acceptable exposure times in hot work environments: criteria were reviewed and updated concerning the maximum increase in core temperature and the acceptable water loss, for acclimatised and nonacclimatised subjects. These limits are intended to protect 95% of the population. Measuring strategy: a strategy was developed to assess the risks in any working situation with varying conditions of climate, metabolic rate or clothing. A detailed methodology was developed in three stages: an "observation" method for the recognition of the conditions that might lead to thermal stress; an "analysis" method for evaluating the problem and optimising the solutions; and an "expert" method for in-depth analysis of the working situation when needed. the different results were used to prepare a revision of the interpretation procedure proposed in the ISO standard 7933. We validated the modified approaches using the database. This involved the whole range of conditions for which the model was extended, namely conditions with high and low radiation, humidity and air velocity as well as fluctuating conditions. Based on these results, the predicted heat strain model was developed: it is presently proposed as an ISO and CEN standard.


Ti-mediated direct and highly stereoselective Mannich reactions between esters and oxime ethers.
The first general method of direct and highly stereoselective Ti-mediated Mannich reaction between three types of simpleesters and E and Z mixtures of oxime ethers (aliphatic and aromatic) is accomplished.


The structural motifs of mineralized hard tissues from nano- to mesoscale: A future perspective for material science.
Biological tissues have developed structures that fulfil their various specific requirements. Mineralized tissues, such as tooth and bone, are often of mechanical competence for load bearing. Tooth enamel is the hardest and toughest mineralized tissue. Despite a few millimeters thick and with minimal regenerative capacity, human tooth enamel maintains its functions throughout a lifetime. Bone provides skeletal support and essential metabolism to our body. Degenerative diseases and ageing induce the loss of mechanical integrity of the bone, increasing the susceptibility to fractures. Tooth and bone share certain commonalities in chemical components and material characteristics, both consisting of nanocrystalline apatite and matrix proteins as their basic foundational structural units. Although the mechanical properties of such mineralized hard tissues remain unclear, it is plausible that they have an inherent toughening mechanism. Nanoindentation is able to characterize the mechanical properties of tooth enamel and bone at multiscale levels, and the results suggest that such toughening mechanisms of enamel and bone may be mainly associated with the smallest-scale structure-function relationships. These findings will benefit the development of advanced biomaterials in the field of material science and will further our understanding of degenerative bone disease in the clinical community.


Transvection effects involving DNA methylation during meiosis in the mouse.
High efficiencies of recombination between LoxP elements were initially recorded when the Cre recombinase was expressed in meiotic spermatocytes. However, it was unexpectedly found that LoxP recombination fell to very low values at the second generation of mice expressing Cre during meiosis. The inability of the LoxP elements to serve as recombination substrates was correlated with cytosine methylation, initially in LoxP and transgene sequences, but later extending for distances of at least several kilobases into chromosomal sequences. It also affected the allelic locus, implying a transfer of structural information between alleles similar to the transvection phenomenon described in Drosophila. Once initiated following Cre-LoxP interaction, neither cis-extension nor transvection of the methylated state required the continuous expression of Cre, as they occurred both in germinal and somatic cells and in the fraction of the offspring that had not inherited the Sycp1-Cre transgene. Therefore, these processes depend on a physiological mechanism of establishment and extension of an epigenetic state, for which they provide an experimental model.


Does decreased fibrinolysis have a role to play in the development of non-neoplastic portal vein thrombosis in patients with hepatic cirrhosis?
Hepatic cirrhosis is characterized by complex abnormalities of the fibrinolytic system. Little is known about the possible association between these alterations and thrombosis. The aim of this study was to evaluate the fibrinolytic profile in cirrhotic individuals with and without portal vein thrombosis (PVT). We measured thrombin activatable fibrinolysis inhibitor (TAFI), total amount of activated TAFI (TAFIa/ai), plasminogen activator inhibitor (PAI-1), plasminogen and fibrinogen plasma levels in 66 cirrhotic patients (33 with and 33 without PVT) and in 66 healthy volunteers. TAFI plasma levels (median [range]) were significantly lower in cirrhotic individuals (5.6 μg/ml [1.7-11.7]) than in controls (10.1 μg/ml [6.6-14.2], p < 0.0001), while TAFIa/ai levels were significantly higher in cases (18.3 ng/ml [0.3-35.4]) than in controls (15.9 ng/ml [7.4-41], p = 0.02). Cirrhotic patients with PVT had higher TAFI (6.6 μg/ml [2.9-10.1]), TAFIa/ai (19.2 ng/ml [11.6-35.4]) and PAI-1 (33.1 ng/ml [27.6-56.3]) plasma levels than those without PVT (3.9 μg/ml [1.7-11.7], p = 0.001; 15.6 ng/ml [10.3-33.9], p = 0.037; 15.9 ng/ml [2.5-29.1], p = 0.004. The fibrinolytic profile in cirrhotic individuals with PVT is characterized by higher levels of TAFI, TAFIa/ai and PAI-1 than in those without PVT. These alterations identify a hypofibrinolytic condition that may increase the risk of developing a thrombotic event.


The added value of non-contrast 3-Tesla MRI for the pre-operative localization of hyperparathyroidism.
We investigated the efficacy of non-contrast 3-Tesla MR imaging added to the combination of sestamibi with99mTc (MIBI) scintigraphy and Ultrasonography (US) for the pre-operative localization of Primary Hyperparathyroidism (PHPT) lesions. A total of 34 parathyroid glands, including nine normal glands, were examined with MIBI, US, and non-contrast 3-Tesla MRI. MRI was performed with the acquisition of T1- and T2-weighted images and fat-suppressed T2-weighted images. We calculated the sensitivities of MIBI, US, and the 'additional' MRI, with knowledge of the former two modalities' results. For the diagnosis of PHPT lesions, the sensitivity values of MIBI, US, and additional MRI were 88.0% (22/25), 84.0% (21/25), and 92.0% (23/25), respectively. Normal glands were not visualized with any modality (0/9). One lesion was detected neither with US nor MRI, but only with MIBI, with the limitation that MIBI represented no more than laterality. The two glands not identified in MRI were 4 mm and 6 mm in their size, which are within the range of normal gland's size. Two lesions were not detected with US or MIBI but were visualized with the additional MRI, which indicated that the MRI contributed an 8.0% (2/25) improvement of sensitivity, compared from that of US. Fat-suppressed T2-weighted images were useful in the identification of parathyroid lesions, as these images helped to differentiate between the lesion and the adjacent tissue. Additional non-contrast 3-Tesla MRI was a useful adjunctive tool for localization of PHPT, which improved the sensitivity of the pre-operative localization of PHPT lesions. Fat-suppressed T2-weighted images contributed to their identification. Evidence from a single descriptive or qualitative study.


Cellular metabolism of fluorescent nitroheterocycles.
Since nitroheterocycles are preferentially metabolized and bound in hypoxic cells, we have examined more than 2 dozen nitroheterocycles as potential fluorescent probes for hypoxia. Using flow cytometry, several patterns of cellular fluorescence (CF) have been observed; for most nitroheterocycles, CF was several fold higher for anoxic than for aerobic cells (which was not predicted based on comparison of the fluorescence spectra of parent drug and reduced products). CF gradually increased when cells were exposed to 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2) or to 4-nitroquinoline-1-oxide (4-NQO), and cells remained fluorescent when the drug was washed off. In contrast, cells exposed to trans-5-amino-3-[5-nitro-2-furyl)vinyl-1,2,4-oxadiazole (NFVO) lost fluorescence with a half-time of 60 minutes. Cells exposed to nitrofurazone (NF-7) reached maximum fluorescence within 30 minutes and then lost fluorescence, even in the presence of the drug. Finally, cells exposed to 3-nitropyrene (NP-3) were initially more fluorescent when incubated under aerobic conditions than anoxic conditions; however, after 2 hours in the presence of NP-3, anoxic cells continued to increase in fluorescence while aerobic cells lost fluorescence. Differences in the patterns of cellular accumulation of fluorescent nitroheterocycles were accompanied by differences in the toxicity and metabolism of these drugs. Therefore, chemical studies alone do not allow us to predict the potential of a compound as a hypoxic probe; studies at the cellular level are also essential.


Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Intravenous bolus or infusion of naloxone in opioid overdose.
A short cut review was carried out to establish whether intravenous boluses of naloxone are better than intravenous infusion in opioid overdose. Altogether 188 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of this best paper is tabulated. A clinical bottom line is stated.


A Multicenter Clinical Study of Single-Kidney Transplantation vs En Bloc Transplantation with Kidneys from Deceased Pediatric Donors.
There are still disputes regarding the choice of surgical approach to harvest organs from pediatric donors for organ recipients. The primary goal of this multicenter, retrospective analysis was to compare outcomes between single-kidney transplantation (SKT) and en bloc kidney transplantation (EBKT). Data from donors and recipients aged 4 to 18 months from 3 transplant centers over 5 year were collected to compare postoperative complications and recoveries of renal function between SKT and EBKT and to determine whether there is a difference in the 1-year patient and kidney survival rate between the 2 groups. Between the SKT and EBKT groups, the incidence of delayed graft function was significantly higher in the SKT group than in the EBKT group (44.1% vs 17.3%, P = .03), and there were no significant differences in other complications (47.0% vs 59.0%, P = .36). Moreover, no significant differences were observed for the overall patient survival rate (P = .08) or the overall graft survival rate (P = .71). The short-term effects of SKT make it worthy of consideration. For infants aged 4 to 18 months, SKT can provide good results, alleviating the current tense situation in kidney donation.


Production of interferon gamma and interleukin 17A in chicken T-cell subpopulations hallmarks the stimulation with live, irradiated and killed avian pathogenic Escherichia coli.
Avian pathogenic Escherichia coli (APEC) causes colibacillosis with different clinical manifestations. The disease is associated with compromised animal welfare and results in substantial economic losses in poultry production worldwide. So far, immunological mechanisms of protection against colibacillosis are not comprehensively resolved. Therefore, the present study aimed to use an ex vivo model applying chicken mononuclear cells stimulated by live and inactivated APEC. For this purpose, an 8-color flow cytometry panel was set up to target viable chicken immune cells including CD45+, CD8α+, CD4+, TCR-γδ+, Bu-1+ cells and monocytes/macrophages along with the cytokines interferon gamma (IFN-γ) or interleukin 17A (IL-17A). The 8-color flow cytometry panel was applied to investigate the effect of live and two different types of inactivated APEC (formalin-killed APEC and irradiated APEC) on the cellular immune response. For that, mononuclear cells from spleen, lung and blood of 10-week-old specific pathogen-free layer birds were isolated and stimulated with live, irradiated or killed APEC. Intracellular cytokine staining and RT-qPCR assays were applied for the detection of IFN-γ and IL-17A protein level, as well as at mRNA level for spleenocytes. Ex vivo stimulation of isolated splenocytes, lung and peripheral blood mononuclear cells (PBMCs) from chickens with live, irradiated or killed APEC showed an increasing number of IFN-γ and IL-17A producing cells at protein and mRNA level. Phenotyping of the cells from blood and organs revealed that IFN-γ and IL-17A were mainly produced by CD8α+, TCR-γδ+ T cells as well as CD4+ T cells following stimulation with APEC. Expression level of cytokines were very similar following stimulation with live and irradiated APEC but lower when killed APEC were applied. Consequently, in the present study, an ex vivo model using mononuclear cells of chickens was applied to investigate the cellular immune response against APEC. The results suggest the relevance of IFN-γ and IL-17A production in different immune cells following APEC infection in chickens which needs to be further investigated in APEC primed birds.


The "comparative growth assay": examining the interplay of anti-cancer agents with cells carrying single gene alterations.
We have developed a "comparative growth assay" that complements current assays of drug effects based on cytotoxicity. A co-culture of two cell lines, one of which is fluorescently labeled, is exposed to a cytotoxic agent and the proportion of fluorescent cells is compared with that of a baseline unexposed co-culture. For demonstration purposes, two HCT116 cell lines (an hMLH1 homozygous and an hMLH1 heterozygous mutant), altered by insertion of vector alone or the same vector carrying an insert for the expression of enhanced green fluorescent protein (EGFP), were exposed to numerous "anti-cancer" agents. The assay was further validated in a system of two cell lines differing only in the expression of the breast cancer resistance protein (BRCP). The assay allowed the estimation of the duration of action of a particular agent. Assessment of the agent's differential activity over a given time in culture could be expressed as a selection rate, which we chose to describe on an "average selection per day" basis. We conclude that this assay: 1) provides insight into the differential dynamic effects of chemotherapeutic agents or radiation; and 2) allows, through the use of matched cell lines, the investigation of critical physiologic features that govern cell sensitivity.


Amino acid modulation of glucose-induced insulin and glucagon release in diabetic patients.
In order to determine whether amino acids have a beneficial effect on glucose tolerance in diabetes, the effect of intravenous infusion of mixed amino acids on plasma insulin, glucagon, and blood glucose responses to oral glucose loading was studied in patients with mild to moderate diabetes. Intravenous infusion of mixed amino acids over a period of 30 min which was started 30 min or immediately before oral glucose loading significantly augmented the insulin response but did not improve the blood glucose curve, probably due to excessive glucagon response. However, amino acid infusion over a period of 60 min started immediately before oral glucose loading evoked a sustained rise of plasma insulin associated with a lesser degree of glucagon secretion, thus causing significant improvement of the blood glucose curve.


Recombinant Treponema pallidum protein Tp0768 promotes proinflammatory cytokine secretion of macrophages through ER stress and ROS/NF-κB pathway.
In response to danger signals, macrophages rapidly produce many inflammatory cytokines that trigger the cascade release of inflammatory mediators, leading to tissue damage, which is an important cause of clinical manifestations of syphilis at all stages. However, we still know very little about the specific mechanism of this process. Tp0768 is an infection-stage-dependent antigen that plays an important role in the infection of Treponema pallidum. In this study, we demonstrated that Tp0768 stimulation of macrophages can cause IL-1β, IL-6, and IL-8 mRNA expression levels to increase in a dose- and time-dependent manner. Further research showed that Tp0768 activated ER stress and the ROS/NF-κB pathway in macrophages. Inhibition of ER stress and the ROS/NF-κB pathway inhibited the expression of IL-1β, IL-6, and IL-8 induced by Tp0768. In addition, pretreatment with a PERK pathway inhibitor significantly reduced the expression of the NF-κB and JNK pathways, while also downregulating the expression of IL-1β, IL-6, and IL-8. Tp0768 stimulation can activate IRE1α/XBP-1 signaling and participate in the induction of inflammatory cytokines through the JNK pathway. These findings indicate that Tp0768 promotes the secretion of proinflammatory cytokines IL-1β, IL-6, and IL-8 by macrophages through ER stress and the ROS/NF-κB pathway, which are also involved in the activation of the NF-κB and JNK pathways that are induced by the PERK pathway and activation of IRE1α/XBP-1 signaling. KEY POINTS: • This study found for the first time that the recombinant Treponema pallidum protein Tp0768 promotes the production of IL-1β, IL-6, and IL-8 by macrophages through ER stress. • Recombinant Treponema pallidum protein Tp0768 regulates the ROS/NF-κB pathway through ER stress. • ER stress-related pathway PERK induces the expression of IL-1β, IL-6, and IL-8 by activating the NF-κB pathway and the JNK pathway. • IRE1α can induce the splicing of XBP-1mRNA and activate the JNK pathway.


Possibility of metabolic control of membrane excitation.
In the guinea pig taenia coli, when glycogen is depleted by repeating Ca-induced contracture in excess K solution containing no glucose, the tension cannot be maintained. The decrease in tension is accompanied by reduction of high energy phosphate compounds and oxygen consumption. When substrate is readmitted to the glycogen-depleted preparation in the presence of 2.4 mM Ca and 20 mM K, the first response is hyperpolarization of the membrane and relaxation, and this is followed by depolarization and development of contracture. The latter response is blocked by verapamil, suggesting that energy supply increases the Ca conductance of the plasma membrane. The early response is considered to be due to activation of electrogenic Ca pump, since this is not affected by ouabain as well as removal of Na and K. ATP produced by substrate readmission is probably preferentially utilized for Ca pump activation to reduce the intracellular Ca. The recovery of tension is likely to be brought about by ATP supply not only to the contractile machinery but also to the plasma membrane to remove inactivation of Ca conductance. It is postulated that as the energy source is depleted, energy consumption is automatically limited by suppressing Ca influx, as a self-defence mechanism. Since beta HB is as effective as glucose in the recovery of these processes, and also in the activation of electrogenic Na pump, the metabolic pathway of oxidative phosphorylation alone can support these functions without a contribution of the glycolytic pathway.


[Fan Yu-qi and "Taiyi miraculous moxa roll"].
To explore the manipulation method, the range of indications and the characteristics of acupoint selection for "Taiyi miraculous moxa roll". Apply computer to make retrieval and statistics of the literature, and then induction and analysis were made by manual methods. The "Taiyi miraculous moxa roll" method substitutes moxibustion for acupuncture, with very wide ranges of indications, the most being the thoracico-abdominal diseases, the next being the diseases in the head, and the less being in the limbs and in the lumbodorsal position; so far as the syndrome differentiation is concerned, the most is deficiency syndrome, the next is cold syndrome, wind syndrome, heat syndrome, phlegm syndrome and so on. The most acupoints selected are on the trunk, and the next on the limbs, and then on the head. This method is worth to be popularized.


Medial portal technique for single-bundle anatomical anterior cruciate ligament (ACL) reconstruction.
The aim of the paper is to describe the medial portal technique for anatomical single-bundle anterior cruciate ligament (ACL) reconstruction. Placement of an ACL graft within the anatomical femoral and tibial attachment sites is critical to the success and clinical outcome of ACL reconstruction. Non-anatomical ACL graft placement is the most common technical error leading to recurrent instability following ACL reconstruction. ACL reconstruction has commonly been performed using a transtibial tunnel technique in which the ACL femoral tunnel is drilled through a tibial tunnel positioned in the posterior half of the native ACL tibial attachment site. ACL reconstruction performed using a transtibial tunnel technique often results in a vertical ACL graft, which may fail to control the combined motions of anterior tibial translation and internal tibial rotation which occur during the pivot-shift phenomenon. The inability of a vertically oriented ACL graft to control these combined motions may result in the patient experiencing continued symptoms of instability due to the pivot-shift phenomenon. The medial portal technique in which the ACL femoral tunnel is drilled through an anteromedial or accessory anteromedial portal allows consistent anatomical ACL tunnel placement. This paper describes the advantages of the medial portal technique, indications for the technique, patient positioning, proper portal placement, anatomical femoral and tibial tunnel placement, graft tensioning and fixation.


The home hemodialysis hub: physical infrastructure and integrated governance structure.
An effective home hemodialysis program critically depends on adequate hub facilities and support functions and on transparent and accountable organizational processes. The likelihood of optimal service delivery and patient care will be enhanced by fit-for-purpose facilities and implementation of a well-considered governance structure. In this article, we describe the required accommodation and infrastructure for a home hemodialysis program and a generic organizational structure that will support both patient-facing clinical activities and business processes.


Unexpected changes to the global methane budget over the past 2000 years.
We report a 2000-year Antarctic ice-core record of stable carbon isotope measurements in atmospheric methane (delta13CH4). Large delta13CH4 variations indicate that the methane budget varied unexpectedly during the late preindustrial Holocene (circa 0 to 1700 A.D.). During the first thousand years (0 to 1000 A.D.), delta13CH4 was at least 2 per mil enriched compared to expected values, and during the following 700 years, an about 2 per mil depletion occurred. Our modeled methane source partitioning implies that biomass burning emissions were high from 0 to 1000 A.D. but reduced by almost approximately 40% over the next 700 years. We suggest that both human activities and natural climate change influenced preindustrial biomass burning emissions and that these emissions have been previously understated in late preindustrial Holocene methane budget research.


Annular diffusion denuder for simultaneous removal of gaseous organic compounds and air oxidants during sampling of carbonaceous aerosols.
A specially designed annular diffusion denuder for simultaneous removal of organic gaseous compounds and atmospheric oxidants in carbonaceous aerosol sampling is presented. Various kinds of denuder coatings were compared with respect to the collection efficiency of both organic gaseous compounds and NO(2) and ozone. The optimum sorbent is a mixture of activated charcoal and sulfite on molecular sieve. To ensure high collection efficiency over long-term field operation, two annular diffusion denuders are combined in series. The first half of the first denuder is filled with Na(2)SO(3) on molecular sieve (23 cm long layer) while the second half of the first denuder and the whole second denuder are filled with activated charcoal (the total length of the charcoal section is 67 cm). At a flow rate of 16.6 L min(-1), the collection efficiency of organic gaseous compounds and atmospheric oxidants in the annular diffusion denuder is better than 95%. Only small losses of aerosol particles (<3.6% in number concentration) were observed in the size range 0.12-2.26 μm. The annular diffusion denuder is compatible with the collection of aerosols on 47-mm diameter quartz fiber filters at a flow rate of 16.6 L min(-1). The use of this denuder enables one to sample carbonaceous aerosols on filters without positive sampling artefacts from volatile organic compounds and interferences from atmospheric oxidants. The annular diffusion denuder has been applied successfully for the sampling of carbonaceous aerosols during field campaigns of typically 1 month each at urban and forested sites in Europe.


Opium and cardiovascular health: A devil or an angel?
Opioids have the highest rate of illicit drug consumption after cannabis worldwide. Opium, after tobacco, is still the most commonly abused substance in the Middle East. In addition to the ease of availability, one reason for the high consumption of opium in Asian countries might be a traditional belief among Eastern people and even medical staff that opium may have ameliorating effects on cardiovascular diseases (CVDs) as well as diabetes mellitus, hypertension, and dyslipidemia. Over the last decade, many studies have been performed on humans and animals to evaluate the interplay between opium consumption and stable coronary artery disease, acute coronary syndromes, and atherosclerosis. In this review, we conclude that opium consumption should be considered a risk factor for CVDs. Healthy individuals, as well as cardiac and diabetic patients, should be informed and educated about the hazardous effects of opium consumption on cardiovascular and other chronic diseases.


Does Early Ureteroneocystostomy After Iatrogenic Ureteral Injury Jeopardize Outcome?
To compare the outcomes of patients who underwent early repair (≤7 days) of iatrogenic ureteral injury with ureteroneocystostomy and compare them to those repaired in a delayed fashion (>7 days). A retrospective review of billing data between 2012 and 2018 identified patients who underwent ureteroneocystostomy for a benign ureteral disease. Inclusion criteria included all ureteral injuries related to a laparoscopic, robotic, or open surgical injury. Patients with ureteral injury related to radiation, stones, or reconstructive surgery were excluded. Patients undergoing reconstruction during the initial injury or within ≤7 days were designated as having undergone early repair, while the remaining were considered delayed repair. Demographics, as well as inpatient and postdischarge data were acquired, and statistical analysis was performed comparing the 2 groups. Sixty-seven patients met inclusion criteria. Early repair was performed on 12 patients, while 55 underwent delayed repair. No significant difference in age, gender, Charlson Comorbidity Score, laterality, stricture location, or history of pelvic/abdominal radiation was noted. Inpatient complications were significantly higher in the immediate group (58 vs 18%, P =.004). Thirty- and 90-day complications were similar. Two patients in the delayed group and none in the immediate group demonstrated stricture recurrence (P =.710). A higher rate of Boari flap ureteral reconstruction was performed in the delayed cohort (P =.001). In this cohort, there was no detectable difference in outcomes when comparing early and delayed ureteroneocystostomy for iatrogenic ureteral injuries.


Osteosarcoma of the hand: a rare case for radiographic appearance, location, and age.
This case describes the history, process of correct diagnosis, and treatment of a patient with pain in her left fourth finger, later found to be an osteosarcoma. A 28-year-old woman presented with pain in her left fourth finger, which she also noticed had been "getting bigger." Moderate tenderness and decreased range of motion were noticed over the metacarpophalangeal joint of the involved finger. Initial radiographs indicated a benign tumor with a pathological fracture, and the patient was referred to a hand surgeon. After debridement of the area, the pathology report returned with a diagnosis of osteoblastic osteogenic osteosarcoma. The patient then underwent amputation of the fourth ray and chemotherapy. This case presents a rare bone tumor of the hand and highlights the importance of clinical decision-making to lead to appropriate patient care.