Effect of ethyl-alpha-hydroxymethylacrylate on selected properties of copolymers and ACP resin composites.
There is an increased interest in the development of bioactive polymeric dental composites and related materials that have potential for mineralized tissue regeneration and preservation. This study explores how the substitution of ethyl alpha-hydroxymethylacryate (EHMA) for 2-hydroxyethyl methacrylate (HEMA) in photo-activated 2,2-bis[p-(2'-hydroxy-3'-methacryloxypropoxy)phenyl]propane (Bis-GMA) and Bis-GMA/tri(ethylene glycol) dimethacrylate (TEGDMA) resins affected selected physicochemical properties of the polymers and their amorphous calcium phosphate (ACP) composites. Rate of polymerization and the degree of conversion (DC) of polymers {EHMA (E), HEMA (H), Bis-GMA/EHMA (BE), Bis-GMA/HEMA (BH), Bis-GMA/TEGDMA/EHMA (BTE) and Bis-GMA/TEGDMA/HEMA (BTH)} were assessed by photo-differential scanning calorimetry and Fourier-Transform Infrared (FTIR) spectroscopy. ACP/BTE and ACP/BTH composites were evaluated for DC, biaxial flexure strength (BFS), water sorption (WS) and mineral ion release. Mid-FTIR and near-IR measurements revealed the following order of decreasing DC: [E, H polymers (97.0%)] > [BE copolymer (89.9%)] > [BH copolymer (86.2%)] > [BTE, BTH copolymers (85.5%)] > [ACP/BTH composite (82.6%)] > [ACP/BTE composite (79.3%)]. Compared to HEMA, EHMA did not adversely affect the BFS of its copolymers and/or ACP composites. Lower WS of BTE copolymers and composites (28% and 14%, respectively, compared to the BTH copolymers and composites) only marginal reduced the ion release from ACP/BTE composites compared to ACP/BTH composites. More hydrophobic ACP composites with acceptable ion-releasing properties were developed by substituting the less hydrophilic EHMA for HEMA.


Development of a checklist for documenting team and collaborative behaviors during multidisciplinary bedside rounds.
The objective of this study was to develop a reliable and valid checklist for documenting team and collaborative behaviors occurring during multidisciplinary bedside rounds. Teamwork and collaboration are important for providing high-quality patient care, yet there are no objective means of evaluating the occurrence of team and collaborative behaviors during bedside rounds. A checklist was developed and tested on 3 general medical units. Items on the checklist were derived from the literature and our medical center's patient-family-centered values. The final version of the checklist was determined to be reliable, valid, and easy to use in the clinical setting. Clinicians, administrators, and investigators are encouraged to use and/or modify this checklist for use in their setting. Further research identifying instruments to objectively measure teamwork and collaboration is needed.


Both LmDicer-1 and two LmDicer-2s participate in siRNA-mediated RNAi pathway and contribute to high gene silencing efficiency in Locusta migratoria.
Dicers belong to a class of large RNase III multidomain ribonucleases and are central components of the RNA interference (RNAi) pathways. In insects, Dicer-2 has been known to cleave long double-stranded RNA (dsRNA) in small interfering RNA (siRNA)-mediated-RNAi pathway. However, Dicer-1 is responsible for cleaving precursor microRNAs (pre28 miRNAs) in miRNA-mediated RNAi pathway. In this study, we identified one LmDicer-1 and two LmDicer-2 (LmDicer-2a and LmDicer-2b) genes in Locusta migratoria. The RNAi of RNAi assay showed that knockdown of each of the Dicer genes reduced RNAi efficiency against a target gene (Lmβ-Tubulin), suggesting that all these genes participated in the siRNA-mediated RNAi pathway. Sequence analyses of the siRNAs generated from dsLmβ-Tubulin after silencing each LmDicer gene showed no significant difference in the pattern of siRNAs mapped to dsLmβ-Tubulin. This result indicated that all the three LmDicers are capable of generating siRNAs from the dsRNA. We then generated recombinant proteins consisting of different domains using Escherichia coli expression system and incubated each recombinant protein with dsLmβ-Tubulin. We found that the recombinant Dicer proteins successfully cleaved dsLmβ-Tubulin. However, LmDicer-2a-R lacking dsRBD domain lost activity, suggesting that dsRBD domain is critical for Dicer function. Furthermore, overexpression of these proteins in Drosophila S2 cells improved RNAi efficiency. Our siRNA affinity chromatography and LC-MS/MS analysis identified LmDicer-2a, LmDicer-2b, LmR2D2, LmAgo2a, LmAgo1, LmStaufen and LmTARBP2 as constituents of RNA-induced silencing complex. Taken together, these data show that both LmDicer-1 and two LmDicer-2s all participate in siRNA-mediated RNAi pathway and likely contribute to high RNAi efficiency in L. migratoria.


The migration, transformation, and risk assessment of heavy metals in residue and bio-oil obtained by the liquefaction of pig manure.
The total contents and chemical speciation analysis of Zn, Cu, Pb, Cr, Mn, Ni, Cd, and As in pig manure (PM), liquefaction residues (LRs), and bio-oils (BOs) derived from PM by liquefaction with ethanol as a solvent at 180-300 °C were thoroughly investigated in this study. The environment risk assessment, leachability, and bioavailability of heavy metals in PM and LRs were studied. The results showed that more than 75% of heavy metals remained in LRs. The total contents of heavy metals in LRs were markedly elevated, but those in BOs gradually decreased with the increase in liquefaction temperature. Moreover, the acid soluble/exchangeable fraction and reducible fraction (F1 + F2) of heavy metals in LRs and BOs was significantly reduced, while oxidizable fraction and stable fraction (F3 + F4) desirably increased after liquefaction. Furthermore, the potential risk of heavy metals in LRs was decreased in comparison to that in PM, but the risk of Pb, Mn, and As had not been obviously reduced; therefore, the LRs from the liquefaction of PM should be pretreated before recycling. Temperatures from 220 to 260 °C were the optimum conditions for disposing of PM by liquefaction with ethanol.


[Comparison of early management of central retinal vein occlusion with ranibizumab versus hemodilution].
This study was designed to evaluate and compare the efficacy of early treatment of CRVO with either hemodilution by erythrocytopheresis or intravitreal (IVT) ranibizumab, or both. A multicentric prospective randomized study including patients with CRVO for less than 1 month was designed. Patients were randomized into 3 treatment groups: hemodilution (HD group), 3 monthly intravitreal injections of ranibizumab followed by PRN treatment (IVT group), or combined treatment (IVT+HD group). A monthly evaluation during a 6-month follow-up included best-corrected visual acuity (BCVA) and macular thickness measurements with OCT. Fluorescein angiography was performed at baseline, month 2 and month 6. Local and systemic tolerability of the different treatments were also compared. Forty-four CRVO patients were included between February 2010 and June 2013: 20 in the IVT group, 13 in the HD group and 11 in the HD+IVT group. The mean duration of CRVO at baseline was 10 days and 16 days at the time of treatment without any significant difference between groups. Retinal ischemia was present at baseline in 40% of eyes in each group. After a 6-month follow-up, no difference between the 3 groups was observed in BCVA (10.5 ETDRS letters, 14.6 and 14.1 in the IVT group, HD group and IVT+HD group respectively, P=0.726) or in macular thickness (398 μ, 440 μ and 379 μ respectively, P=0.465). The time until treatment from CRVO onset, ranging from 1 to 35 days, was not correlated to final outcomes. No significant difference in the mean number of IVT (3.2 in the IVT+HD group vs 3.7 in the IVT group) was observed at 6 months. No difference in BCVA nor in macular thickness was seen at M6 between the study groups. The duration of CRVO at the time of the initiation of the treatment was not correlated to better visual outcomes. Therefore, etiologic treatment with HD can still be proposed as a first-line treatment in young patients, which allows delaying or avoiding the IVT treatment and its potential side effects. Anti-VEGF IVT still remains an effective option in every case and can be started one month after the beginning of the CRVO.


Prediction of hand strength by hand injury severity scoring system in hand injured patients.
Hand strength is a key component in human hand function, especially to those with strength-required jobs. To evaluate post-injury hand strength, this study was conducted to assess the predictability of hand injury severity scoring (HISS) system to hand strength after a period of recovery. In this retrospective study, 80 traumatic hand-injured subjects were recruited. Hand strength tests measured by dynamometers were performed by subjects in ways of palmar pinch, lateral pinch and cylindrical grip. The logistic regression models were used to evaluate the risk of poor hand strength among the severities represented by HISS scores. A positive correlation exists between the differences of the strength of two hands, and the severity shown by HISS system in the poorer half of subjects. The risk to be in poorer half of palmar pinch subtest is significant in moderate subgroup of HISS severity. The risk to be in poorer half of cylindrical grip subtest is significant in major subgroup of HISS severity. The risk to be in poorer half of palmar pinch and cylindrical grip subtests is significant in skeletal component of HISS system. HISS is a descriptive severity scoring system to hand injury and also a useful instrument to predict functional outcome. This study revealed the HISS system may predict post-injury hand strength after recovery.


Resveratrol loaded nanoparticles attenuate cognitive impairment and inflammatory markers in PTZ-induced kindled mice.
Resveratrol has been found to exert protective effects in neurological disorders, including epilepsy. However, its poor bioavailability and difficulty in reaching the brain's targeted location reduce resveratrol's efficacy substantially. The side effects due to the higher concentration of drugs are another matter of concern. The objective of the present study is to propose solutions to these issues by encapsulating resveratrol in glutathione-coated collagen nanoparticles' core. The collagen nanoparticles increase the resveratrol's bioavailability, and glutathione helps in the passage of the encapsulated resveratrol to the target location in the brain. The concentration also substantially reduces due to resveratrol's encapsulation in glutathione-coated collagen nanoparticles. The encapsulated resveratrol is termed nanoresveratrol. The effectiveness of nanoresveratrol on epilepsy seizures was evaluated through histopathological examinations, ELISA tests, and qRT-PCR tests on the hippocampus of the kindled mice. The novelty of the present study thus lies in (i) the synthesis of nanoresveratrol using glutathione-coated collagen nanoparticles and (ii) the application of synthesized nanoresveratrol in the treatment of epilepsy. The study's outcome shows that nanoresveratrol has a favorable impact in reducing cognitive impairment in kindled mice, and it is more effective in controlling epilepsy seizures than resveratrol. The p-values of all the nanoresveratrol-given groups of mice (compared with the diseased group) were substantially smaller (∼10-4 to 10-2) than the significance level (0.05), indicating that the nanoresveratrol-given groups are significantly different from the diseased group, i.e., the nanoresveratrol has a significant effect on the mice. The concentration of resveratrol also decreases substantially in the proposed nanoformulation. It was observed that even 0.4 mg/kg of nanoformulation of resveratrol is performing better than 40 mg/kg of resveratrol.


Histone deacetylases and their role in asthma.
The aim of this article is to provide an overview of the classical histone deacetylase (HDAC) enzymes and HDAC inhibitors. The discussion is focused on the potential anti-asthmatic effects of this group of compounds. Medline was used with the search terms, "asthma and HDAC," "asthma and Trichostatin A," "asthma and valproic acid," "allergic airways disease and HDAC," "allergic airways disease and Trichostatin A," and "allergic airways disease and valproic acid." Manuscripts from the past decade were accessed. Historical literature dating from the 1960s was accessed for the use of anti-epileptics in the treatment of asthma. Preliminary clinical trials with anti-epileptic drugs including the well-known HDAC inhibitor, valproic acid, have shown long-lasting anti-asthmatic effects providing the basis for the evaluation of this class of compounds in asthma. Studies using the prototypical HDAC inhibitor, Trichostatin A, in well-established murine models of allergic airways disease have also indicated beneficial effects. Although the precise mechanisms are still controversial, inhibition of airway hyperresponsiveness and agonist-induced contraction as well as anti-inflammatory effects have been described for HDAC inhibitors in asthma.


Acoustic radiation force impulse sonography in assessing children with biliary atresia for liver transplantation.
Children with biliary atresia are prone to developing progressive hepatic fibrosis and biliary cirrhosis following the Kasai operation. The only treatment is liver transplantation. To assess liver fibrosis by acoustic radiation force impulse elastography (ARFI) in children who had Kasai operation, with the goal of identifying an ARFI value cut-off for children requiring liver transplantation. Of the 32 post-Kasai children included, 19 were transplanted or listed for transplantation (group A), while 13 were not on the list during their follow-up (group B). We recorded biopsies, blood samples and ARFI values over time, including at Kasai operation and at transplantation. We estimated an association between groups and continuous variables using generalized estimating equations, and we compared categorical variables using the Fisher exact test. Portal hypertension signs were similar in both groups, whereas ARFI values were higher in group A (mean±standard deviation=3.3±1.2 m/s) than in group B (2.0±0.7 m/s; P=.0003). Eighteen of 19 (94.7%) children in group A and 6/13 (46.2%) children in group B presented with two consecutive ARFI values ≥2 m/s (sensitivity=7%, specificity=53.8%; P=0.003). We found that children who were transplanted had two consecutive ARFI values ≥2 m/s during follow-up. ARFI for evaluation of post-Kasai liver fibrosis may assist the long-term assessment of biliary atresia and may even guide treatment decisions.


Inhibition of Fatty Acid Amide Hydrolase (FAAH) During Adolescence and Exposure to Early Life Stress may Exacerbate Depression-like Behaviors in Male and Female Rats.
Early-life stress (ELS) is associated with later onset of depression. Early cannabis use may be a risk factor that interacts with environmental factors to increase the risk of psychopathologies. We aimed to examine the long-term effects of ELS on depression- and anxiety-like behavior, and examine whether chronic fatty acid amide hydrolase (FAAH) inhibition during mid-adolescence could ameliorate or exacerbate ELS effects on behavior. Male and female rats were exposed to ELS during post-natal days (P) 7-14, injected with the FAAH inhibitor URB597 (0.4 mg/kg, i.p.) or vehicle for 2 weeks during mid-adolescence (P30-45) or late-adolescence (P45-60). Rats were tested in adulthood for behavior and alterations in CB1 receptors (CB1r) and glucocorticoid receptors (GRs) in the brains' stress circuit. ELS produced decreased social preference, impaired social recognition, increased learned helplessness and anxiety-like behavior. Administering URB597 during mid-adolescence did not prevent the deleterious long-term effects of ELS on behavior in males and females. When URB597 was administered during late-adolescence, it ameliorated ELS-induced depression- and anxiety-like behavior. Moreover, in males, ELS and URB597 decreased CB1r levels in the prefrontal cortex (PFC) and CA1 and GRs in the PFC and basolateral amygdala (BLA). In females, ELS and URB decreased CB1r in the BLA and GRs in the CA1 and BLA. The findings suggest that mid-adolescence, as opposed to late-adolescence, may not be a potential developmental period for chronic treatment with FAAH inhibitors and that sex-dependent alterations in CB1r and GRs expression in the BLA-PFC-CA1 circuit may contribute to the depressive behavioral phenotype.


The Reliability of Strength Tests Performed In Elevated Shoulder Positions Using a Handheld Dynamometer.
The reliable measurement of shoulder strength is important when assessing athletes involved in overhead activities. Swimmers' shoulders are subject to repetitive humeral elevation and consequently have a high risk of developing movement-control issues and pain. Shoulder-strength tests performed in positions of elevation assist with the detection of strength deficits that may affect injury and performance. The reliability of isometric strength tests performed in positions of humeral elevation without manual stabilization, which is a typical clinical scenario, has not been established. To establish the relative and absolute intrarater reliability of shoulder-strength tests functional to swimming in 3 body positions commonly used in the clinical setting. Repeated-measures reliability study. Research laboratory. 15 university students and staff (mean ± SD age 24 ± 8.2 y). Isometric shoulder-strength tests were performed in positions of humeral elevation (flexion and extension in 140° abduction in the scapular plane, internal and external rotation in 90° abduction) on subjects without shoulder pain in supine, prone, and sitting. Subjects were tested by 1 examiner with a handheld dynamometer and retested after 48 h. Relative reliability (ICC3,1) values with 95% CI. Absolute reliability was reported by minimal detectable change (MDC). Good to excellent intrarater reliability was found for all shoulder-strength tests (ICC .87-.99). Intrarater reliability was not affected by body position. MDC% was <16% for every test and ≤11% for tests performed in supine. Shoulder flexion, extension, and internal- and external-rotation strength tests performed in humeral elevation demonstrated excellent to good intrarater reliability regardless of body position. A strength change of more than 15% in any position can be considered meaningful.


Forced expression of indoleamine-2,3-dioxygenase in human umbilical cord-derived mesenchymal stem cells abolishes their anti-apoptotic effect on leukemia cell lines in vitro.
The ability of mesenchymal stem cells (MSCs) to preserve cancer cells potentially constitutes the adverse effect of MSC-based cell therapy in the context of hematologic malignancy. In an effort to reverse this undesirable feature of MSCs, we manipulated human umbilical cord-derived MSCs (UC-MSCs) to express indoleamine-2,3-dioxygenase (IDO), an enzyme that induces immune suppression by inhibiting T cell proliferation and triggering apoptosis in immune cells. Cultures of human UC-MSCs were generated by plastic adherence method. Full-length cDNA of human IDO was cloned into adenovirus shuttle vector. Then, the recombinant virus harboring IDO gene was produced in 293 cells and used to infect UC-MSCs. Expression of IDO protein was detected within infected UC-MSCs, and accumulation of kynurenine was observed in the supernatant. Two human leukemia cell lines, Jurkat and HL-60, were cultured on the monolayer of native or infected UC-MSCs, respectively. It was observed that forced IDO expression abolished the anti-apoptotic effect of UC-MSCs on these leukemia cells and enhanced their proliferation inhibitory effect on activated human lymphocytes as well as leukemia cells. These results suggested that equipping MSCs with IDO could be one of the reasonable strategies to reverse their cancer-supportive effect unfavorable for clinical applications.


Refractory and new-onset diabetes more than 5 years after gastric bypass for morbid obesity.
Few studies about long-term glucose homeostasis in bariatric patients are available. In a prospective protocol that included retrospective information, outcome of patients with both impaired and normal fasting blood glucose (FBG) was monitored to assess frequencies and correlates. Patients submitted to Roux-en-Y gastric bypass were classified as group I, elevated FBG, and group II, normal controls. Those in group I with improvement in FBG were defined as responsive and the others as refractory. Group II participants progressing to new-onset diabetes (NOD) or prediabetes represented NOD cases; the remaining were listed as stable controls. FBG was the main endpoint, but HbA1c results were considered, along with diet composition and general biochemical profile. Among 97 selected patients, 51 belonged to group I (52.4 ± 10.5 years, 29.6 % males, initial body mass index (BMI) 58.4 ± 13.4, current BMI 35.1 ± 8.4 kg/m(2)) and 46 to group II (48.2 ± 10.5 years, 19.6 % males, initial BMI 55.5 ± 8.8, current BMI 33.9 ± 6.9 kg/m(2)). Follow-up was 7-9 years, and 31.4 % (16/51) of group I were classified as refractory, whereas 15.2 % (7/46) of the controls converted to NOD. Multivariate analysis pointed out higher current BMI, older age, consumption of antidiabetic drugs, and male gender as features of refractory cases, whereas NOD participants were not significantly different from non-progressing controls. This is the first investigation, to the best of our knowledge, to underscore the correlates of refractory and NOD within the bariatric context. Further studies are recommended as such information could be valuable for patient selection, prognostic scoring, and outcome monitoring.


Multimorbidity and patient-reported diagnostic errors in the primary care setting: multicentre cross-sectional study in Japan.
There is lack of evidence for the association between multimorbidity and diagnostic errors. Information on diagnostic errors from patients' perspectives is crucial to improve the diagnostic process. In this study, we aimed to investigate patient-reported diagnostic errors and to examine the relationship between multimorbidity and patient-reported diagnostic errors in the primary care setting. Multicentre cross-sectional study. A primary care practice-based research network in Japan (25 primary care facilities). Adult outpatients filled out a standardised questionnaire. Patient-reported diagnostic errors. Data collected from 1474 primary care outpatients were analysed. The number of participants who reported diagnostic errors was 57 (3.9%). Most of the missed diagnoses were common conditions in primary care, such as cancer, dermatitis and hypertension. After adjustment for possible confounders and clustering within facilities, multimorbidity was positively associated with patient-reported diagnostic errors (adjusted OR=1.83, 95% CI 1.01 to 3.31). The results of the sensitivity analysis were consistent with those of the primary analysis. The present study showed a lower proportion of patients reporting experiences of diagnostic errors in primary care than those reported in previous studies in other countries. However, patients with multimorbidity are more likely to report diagnostic errors in primary care; thus, further research is necessary to improve the diagnostic process for patients with multimorbidity.


Effect of in vitro cultivation and Mycoplasma pneumoniae infection on intracellular cyclic AMP levels in hamster tracheal organ cultures.
Exogenous cyclic AMP and dibutyryl cyclic AMP decreased the relative ciliary activity values of tracheal organ cultures. In contrast, theophylline and cholera toxin were not ciliostatic. The use of a radioimmunoassay for cyclic AMP indicated that all of the tested substances increased intracellular cyclic AMP levels to some extent (from 3-fold for cholera toxin to almost 40-fold for dibutyryl cyclic AMP). Physical inactivation of explants by either freeze-thaw or heat destroyed all ciliary activity and greatly decreased intracellular cyclic AMP levels. Cyclic AMP levels of explants remained relatively constant during in vitro cultivation. Three strains of Mycoplasma pneumoniae were found to contain extremely low amounts of cyclic AMP. Infection of tracheal explants produced a significant decrease in relative ciliary activity, but only a slight decline in organ-culture cyclic AMP levels.


Laparoscopic dual mesh repair of a diaphragmatic hernia of Bochdalek in a symptomatic elderly patient.
Bochdalek hernia is a congenital posterolateral diaphragmatic defect. It is usually diagnosed in newborns and children; the mortality rate is high due to further congenital anomalies such as pulmonary hypoplasia and pulmonary hypertension. In adulthood, the diagnosis is often incidental, while sometimes it is related to gastrointestinal or respiratory symptoms, or to herniated viscera complications. Thus, surgical treatment is advocated. We herein report a case of an 86-year-old woman with dyspnea and dysphagia. After the diagnosis by barium enema and CT scan of the herniated stomach and the greater omentum through a left side foramen of Bochdalek, she underwent a successful laparoscopic dual mesh repair and was discharged on the 6th postoperative day.


Analysis of the histopathology of radiation myelopathy.
An analysis of published histopathology reports of patients with radiation myelopathy was performed. Radiation lesions in the spinal cord were classified as primarily white matter parenchymal lesions (type 1), primarily vascular lesions (type 2), or a combination of vascular and white matter lesions (type 3). The presence or absence of a mononuclear inflammatory reaction was also noted. Type 1 and type 3 lesions had comparable latent periods, both significantly shorter than those observed for type 2 lesions. The anatomical level of the irradiation did not appear to influence the type of lesion. Inflammatory reaction was observed with greater frequency in type 3 lesions. For all types of lesions, the average latent periods in patients with inflammatory reactions were shorter than in those without inflammation. In the cases in which disease status was evaluated, 70% of the patients were free of disease or had no evidence of recurrence at autopsy.


A comparison of the effect of age on levator ani and obturator internus muscle cross-sectional areas and volumes in nulliparous women.
Functional tests have demonstrated minimal loss of vaginal closure force with age. So we tested the null hypotheses that age neither affects the maximum cross-sectional area (CSA) nor the volume of the levator muscle. Corresponding hypotheses were also tested in the adjacent obturator internus muscle, which served as a control for the effect of age on appendicular muscle in these women. Magnetic resonance images of 15 healthy younger (aged 21-25 years) and 12 healthy older nulliparous women (aged >63 years) were selected to avoid the confounding effect of childbirth. Models were created from tracing outlines of the levator ani muscle in the coronal plane, and obturator internus in the axial plane using 3D Slicer v. 3.4. Muscle volumes were calculated using Slicer, while CSA was measured using Imageware™ at nine locations. The hypotheses were tested using repeated measures analysis of variance with P < 0.05 being considered significant. The effect of age did not reach statistical significance for the decrease in levator ani muscle maximum CSA or the decrease in volume (4.3%, P = 0.62 and 10.9%, 0.12, respectively). However, age did significantly adversely decrease obturator internus muscle maximum CSA and volume (24.5% and 28.2%, P < 0.001, respectively). Significant local age-related changes were observed dorsally in both muscles. Unlike the adjacent appendicular muscle, obturator internus, the levator ani muscle in healthy nullipara does not show evidence of significant age-related atrophy.


An insight into the in vivo imaging potential of curcumin analogues as fluorescence probes.
Curcumin and its derivatives have good electrical and optical properties due to the highly symmetric structure of delocalized π electrons. Apart from that, curcumin and its derivatives can interact with numerous molecular targets, thereby exerting less side effects on human body. The fluorescence emission wavelength and fluorescence intensity of curcumin can be enhanced by modifying its π-conjugated system and ß-diketone structure. Some curcumin-based fluorescent probes have been utilized to detect soluble/insoluble amyloid-ß protein, intracranial reactive oxygen species, cysteine, cancer cells, etc. Based on the binding characteristics of curcumin-based fluorescent probes with various target molecules, the factors affecting the fluorescence intensity and emission wavelength of the probes are analyzed, in order to obtain a curcumin probe with higher sensitivity and selectivity. Such an approach will be greatly applicable to in vivo fluorescence imaging.


Transmembrane delivery of polypeptide hormones bypassing the intrinsic cell surface receptors: a conjugate of insulin with alpha 2-macroglobulin (alpha 2M) recognizing both insulin and alpha 2M receptors and its biological activity in relation to endocytic pathways.
125I-labeled insulin has been cross-linked to alpha 2-macroglobulin (alpha 2M) via a disulfide bond. The resulting insulin-alpha 2M conjugate carried 2.2 insulin moieties per mole of alpha 2M and was able to deliver insulin into rat hepatoma cells H35 and HTC. The insulin delivery was mediated predominantly through alpha 2M receptors and 2 h after binding it was found in the lyposomal fractions in the form of conjugate. When the conjugate was applied to rat hepatoma cells it stimulated activity of tyrosine aminotransferase (TAT) with a potency one-half that of native insulin. Hepatoma cells which were treated with conjugate in the presence of bacitracin were also stimulated for TAT activity. Since bacitracin completely inhibited the alpha 2M binding to its receptors, but inhibited conjugate binding by only 80%, this stimulation must have resulted from the remaining binding of conjugate. These results indicate that the insulin-alpha 2M conjugate was biologically active if it bound to insulin receptors, but that the conjugate bound and internalized through alpha 2M receptors did not act as a mediator for TAT activation. Our results using Percoll density gradients indicate a difference in intracellular processing between insulin, alpha 2M and the conjugate. Mechanisms of action of the conjugate are discussed in relation to the receptor-mediated endocytic pathways.


AGRN promotes lung adenocarcinoma progression by activating Notch signaling pathway and acts as a therapeutic target.
Lung cancer is the main reason for cancer-associated death globally, and lung adenocarcinoma (LUAD) is the most prevalent subtype of lung cancer. Recently, AGRN is considered playing an vital role in the development of some cancers. However, the regulatory effects and mechanisms of AGRN in LUAD remain elusive. In this study, we clarified the significant upregulation of AGRN expression in LUAD by single-cell RNA sequencing combined with immunohistochemistry. Besides, we confirmed that LUAD patients with high AGRN expression are more susceptible to lymph node metastases and have a worse prognosis by a retrospective study of 120 LUAD patients. Next, we demonstrated that AGRN directly interact with NOTCH1, which results in the release of the intracellular structural domain of NOTCH1 and the subsequent activation of the NOTCH pathway. Moreover, we also found that AGRN promotes proliferation, migration, invasion, EMT and tumorigenesis of LUAD cells in vitro and in vivo, and that these effects are reversed by blocking the NOTCH pathway. Furthermore, we prepared several antibodies targeting AGRN, and clarify that Anti-AGRN antibody treatment could significantly inhibit proliferation and promote apoptosis of tumor cells. Our study highlights the important role and regulatory mechanism of AGRN in LUAD development and progression, and suggests that antibodies targeting AGRN have therapeutic potential for LUAD. We also provide theoretical and experimental evidence for further development of monoclonal antibodies targeting AGRN.


Shear stress-induced angiogenesis in mouse muscle is independent of the vasodilator mechanism and quickly reversible.
Is modulation of skeletal muscle capillary supply by altering blood flow due to a presumptive shear stress response per se, or dependent on the vasodilator mechanism? The response to four different vasodilators, and cotreatment with blockers of NO and prostaglandin synthesis, was compared. Femoral artery blood flow was correlated with capillary-to-fibre ratio (C:F) and protein levels of putative angiogenic compounds. All vasodilators induced a similar increase in blood flow after 14 days, with a similar effect on C:F (1.62 ± 0.05, 1.60 ± 0.01, 1.57 ± 0.06, 1.57 ± 0.07, respectively, all P < 0.05 vs. control 1.20 ± 0.01). Concomitant inhibitors revealed differential effects on blood flow and angiogenesis, demonstrating that a similar response may have different signalling origins. The time course of this response with the most commonly used vasodilator, prazosin, showed that blood flow increased from 0.40 mL min-1 to 0.61 mL min-1 by 28 days (P < 0.05), dropped within 1 week after the cessation of treatment (0.54 mL min-1 ; P < 0.05) and returned to control levels by 6 weeks. In parallel with FBF, capillary rarefaction began within 1 week (P < 0.05), giving C:F values similar to control by 2 weeks. Of the dominant signalling pathways, prazosin decreased muscle VEGF, but increased its cognate receptor Flk-1 (both P < 0.01); levels of eNOS varied with blood flow (P < 0.05), and Ang-1 initially increased, while its receptor Tie-2 was unchanged, with only modest changes in the antiangiogenic factor TSP-1. Hyperaemia-induced angiogenesis, likely in response to elevated shear stress, is independent of the vasodilator involved, with a rapid induction and quick regression following the stimulus withdrawal.


A two-chain tumour-related suppressor factor specific for a sequence antigen.
Suppressor factors in the serum of CBA mice bearing transplanted methylcholanthrene-induced tumours suppressed the leukocyte adherence inhibition reaction between tumour-sensitized peritoneal cells and a sequenced antigen, myelin basic protein (MBP). Chromatography on Sephacryl S-200 columns demonstrated suppressive activity in low molecular weight fractions (40-50 kDa) derived from whole serum. The factors were absorbed from serum by MBP, but not by lysozyme, coupled to Sepharose gel. Cleavage with sodium dodecyl sulphate (SDS) and absorption with either MBP or anti-I-J antibody on Sepharose gels yielded inactive supernatants that regained suppressive activity when combined. Elution of these gels with acidic buffer yielded eluates that were inactive alone but suppressive when combined. These results indicate that the tumour-related suppressor factors consist of two chains separable by detergent action; one chain is antigen (MBP)-binding and the other contains an I-J determinant.


Expanding aliasing limit in measurement of tissue velocity using autocorrelation method.
Autocorrelation using in-phase and quadrature (IQ) signals suffers from aliasing when the velocity of rapidly moving tissue, such as the heart wall, is measured. In the present study, a simple method was proposed to expand the aliasing limit. In the proposed method, the velocity difference between two successive frames (corresponding to acceleration) of tissue was also estimated directly from IQ signals. When aliasing occurs in the velocity in the current frame, which was estimated from IQ signals, the velocity in the current frame was corrected by adding the velocity difference to the velocity in the previous frame. Using this procedure, the velocity can be estimated if the difference between velocities in the current and previous frames is less than the aliasing limit. The velocity of the posterior heart wall in the longitudinal-axis view of about 0.08 m/s could be estimated under the aliasing limit of the conventional autocorrelation method of 0.047 m/s. Myocardial velocity over the conventional aliasing limit could be measured by the proposed method.


Enumeration of Yersinia enterocolitica O:3 from the porcine oral cavity, and its occurrence on cut surfaces of pig carcasses and the environment in a slaughterhouse.
The number of Yersinia enterocolitica serogroup O:3/biovar 4 recovered from five porcine tongues, ranged from 0-186/cm2, the corresponding range for the tonsils was, 0-1720/cm2. This bacterium was isolated from the oral cavity of 25 (83.3%) of 30 freshly eviscerated slaughter pigs. The isolation rates obtained from the different cut surfaces were: cranial incision, 46.7%; abdominal incision, 43.3%; and circumanal incision, 26.7%. Seven pigs harboured the bacteria in the oral cavity only, and one pig yielded Y. enterocolitica serogroup O:3/biovar 4 only from the abdominal incision. In eighteen pigs the bacteria was obtained from the oral cavity and at least one of the cut surfaces, four of these pigs yielding the bacteria from all sites sampled. Y. enterocolitica O:3/biovar 4 was isolated from samples taken from the following sites in the slaughterhouse: the floor in the bleeding area, the floor in the eviscerating area, the viscera table, and the floor next in line after the meat inspection area. The organism was not found on the floor in the meat inspection area. It is concluded that modern slaughtering technology and the routines followed by meat inspection personnel have probably contributed to the relatively high rates of this bacteria on carcass cut surfaces detected in this study. The results of this investigation thus emphasize the need to consider the introduction of changes in slaughtering technology and meat inspection practices presently being employed in Norwegian slaughterhouses.


Mechanism and Scope of Preferential Enrichment, a Symmetry-Breaking Enantiomeric Resolution Phenomenon.
The mechanism of "preferential enrichment", an unusually symmetry-breaking enantiomericresolution phenomenon that is observed upon simple recrystallization of a certain kind of organic racemiccrystal from the usual organic solvents without any external chiral element, has been rationalized in termsof a complexity system involving multistage processes that affect each other. These processes comprise:(1) preferential homochiral molecular association to give one-dimensional (1D) R andS chains in solution; (2) formation of γ-form prenucleationaggregates consisting of the same homochiral 1D chains; (3) nucleation and crystal growth of the metastableγ-form crystal composed of irregular alignment of the homochiral 1D R andS chains; (4) the solvent-assisted solid-to-solid type of polymorphictransition of the incipient γ-form crystal into the more stable polymorphic form, such as the α-,δ-, or ε-form; and (5) partial crystal disintegration in the transformed crystal to releaseexcess R or S moleculesinto solution. Based on this mechanism, both induction and inhibition of preferential enrichment have beenachieved by controlling the mode of polymorphic transition during crystallization with appropriate seedcrystals. We call this forced polymorphic transition on the surface of a seed crystal "epitaxialtransition".


Plasma volume changes induced by sequential ultrafiltration-hemodialysis and sequential hemodialysis-ultrafiltration.
Isolated ultrafiltration (UF) has been shown to preserve plasma volume (PV) by means of a high plasma refilling rate, mediated by a rapid rise in oncotic pressure. This mechanism contributes to the good tolerance of sequential ultrafiltration-hemodialysis (SUH). This study compared PV changes induced by SUH and sequential hemodialysis-ultrafiltration (SHU). Seven dialysis patients underwent two sets of SUH and SHU, in which 2 h of UF (approximately equal to 3L) respectively preceded or followed 2 h of no-weight-change hemodialysis (ISO HD). VEM (volume of extravascular mobilization), VEM/VUF (percent of plasma refilling rate) and delta PV were calculated by mathematical formulas. Results showed: 1) a high VEM/VUF during the UF period, either before or after ISO HD: 80 and 77% respectively; 2) a significant increase in PV during ISO HD after UF, compared to ISO HD before UF: + 229 ml and + 43 ml, P less than 0.05; 3) VEM/VUF significantly higher during SUH than during SHU: 87 and 80%, P less than 0.01. In conclusion, the decrease in PV was lower in SUH than in SHU (11 vs. 19%, P less than 0.001) because the plasma refilling persisted through the ISO HD period.


[The myosin-activating protein kinases in human myocardium: localization and content].
Using the immunofluorescence approach, we have determined that the recently detected protein kinases, among which are RhoA-activated kinase, integrin-linked kinase, zipper interacting protein kinase, and death-associated protein kinase, which are capable of phosphorylating myosin, are localized in the Z-lines sarcomeres of human myocardium. Additionally, we studied the content of integrin-linked and zipper interacting protein kinases in human embryonic myocardium, as well as in normal and hypertrophic adult human heart. The content of these protein kinases in adult normal myocardium increases in comparison with the embryonic heart. The content of integrin-linked and zipper interacting protein kinases in hypertrophic myocardium is higher compared with the normal adult heart. The data obtained suggest the involvement of these protein kinases in the development and hypertrophy of human heart.


Constitutive internalization and association with adaptor protein-2 of the interleukin-6 signal transducer gp130.
The transmembrane protein gp130 is the common signalling receptor subunit for the interleukin-6 (IL-6)-type cytokines. It has recently been shown that the cytoplasmic domain of gp130 contains a dileucine internalization motif and that endocytosis of gp130 occurs signal-independent. Here, we have studied whether gp130 itself undergoes constitutive internalization or whether its endocytosis is stimulated by formation of the IL-6/IL-6R/gp130 complex. Using two different assays, we found that gp130 is internalized independent from IL-6/IL-6R stimulation. In addition, we show that gp130 is constitutively associated with the cell surface adaptor complex AP-2. Our findings strongly suggest endocytosis of gp130 to be constitutive.


Antimicrobial cotton gauzes modified with poly(acrylic acid-co-maltodextrin) hydrogel using chitosan as crosslinker.
Cotton gauzes were grafted with a hydrogel of maltodextrin (MD) and poly(acrylic acid) (PAAc) using N-maleyl chitosan as crosslinker to obtain materials with antimicrobial properties. Reaction parameters including monomer, crosslinker, and initiator concentrations were studied. The modification with the copolymer poly(acrylic acid)-co-maltodextrin (PAAc-co-MD) was corroborated by Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). The grafted gauzes (gauze-g-(PAAc-co-MD)) were able to load vancomycin and inhibit the growth of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria. In addition, the incorporation of chitosan as crosslinker showed a synergistic effect against these bacteria. The prepared gauze-g-(PAAc-co-MD) materials could be used in the biomedical area particularly as antimicrobial wound dressings.


Enhancement of bone regeneration by coadministration of angiogenic and osteogenic factors using messenger RNA.
Bone defects remain a challenge today. In addition to osteogenic activation, the crucial role of angiogenesis has also gained attention. In particular, vascular endothelial growth factor (VEGF) is likely to play a significant role in bone regeneration, not only to restore blood supply but also to be directly involved in the osteogenic differentiation of mesenchymal stem cells. In this study, to produce additive angiogenic-osteogenic effects in the process of bone regeneration, VEGF and Runt-related transcription factor 2 (Runx2), an essential transcription factor for osteogenic differentiation, were coadministered with messenger RNAs (mRNAs) to bone defects in the rat mandible. The mRNAs encoding VEGF or Runx2 were prepared via in vitro transcription (IVT). Osteogenic differentiation after mRNA transfection was evaluated using primary osteoblast-like cells, followed by an evaluation of the gene expression levels of osteogenic markers. The mRNAs were then administered to a bone defect prepared in the rat mandible using our original cationic polymer-based carrier, the polyplex nanomicelle. The bone regeneration was evaluated by micro-computerized tomography (μCT) imaging, and histologic analyses. Osteogenic markers such as osteocalcin (Ocn) and osteopontin (Opn) were significantly upregulated after mRNA transfection. VEGF mRNA was revealed to have a distinct osteoblastic function similar to that of Runx2 mRNA, and the combined use of the two mRNAs resulted in further upregulation of the markers. After in vivo administration into the bone defect, the two mRNAs induced significant enhancement of bone regeneration with increased bone mineralization. Histological analyses using antibodies against the Cluster of Differentiation 31 protein (CD31), alkaline phosphatase (ALP), or OCN revealed that the mRNAs induced the upregulation of osteogenic markers in the defect, together with increased vessel formation, leading to rapid bone formation. These results demonstrate the feasibility of using mRNA medicines to introduce various therapeutic factors, including transcription factors, into target sites. This study provides valuable information for the development of mRNA therapeutics for tissue engineering.


Dysfunctional feedback processing in adolescent males with conduct disorder.
Abnormalities in neural feedback-processing systems may play a role in the development of dysfunctional behavior in individuals diagnosed with conduct disorder (CD). The present study investigated the relation between CD adolescents and feedback processing by measuring event-related potentials (ERPs) in a single outcome gambling task, which included reward valence (loss and gain) and reward magnitude (10 and 50cents) as outcomes. N2 and P3 components have been established as effective indicators in studies of behavioral disinhibition, reward processing, and decision-making. Eighteen adolescent males (age: 13-17years) diagnosed with CD and 19 healthy age-matched male controls were recruited. Compared to healthy controls, CD individuals exhibited reduced N2 amplitudes in response to loss condition. There was also a significant decreased P3 amplitude in all conditions. The amplitudes of P3 were negatively correlated with impulsivity scores across both groups, and the amplitudes of N2 were positively correlated with impulsivity scores across both groups. Our findings suggest that adolescents with CD may be impaired in neural sensitivity feedback and the processing of environmental cues compared to healthy controls. Moreover, N2 and P3 may be reliable indices to detect different sensitivity in reward and punishment feedback processing.


Association between dental anxiety and intraoperative pain during root canal treatment: a cross-sectional study.
To investigate whether dental anxiety influences the intraoperative pain felt by patients during root canal treatment. In a cross-sectional design, 180 patients (90 men and 90 women) were included. Pre-operative anxiety levels were assessed using the short-form Dental Anxiety Inventory (S-DAI). Pain during root canal treatment was evaluated using a 10-cm visual analogue scale (VAS) that ranked the level of pain between 'Absence of pain' and 'Unbearable pain'. The minimal sample size was determined using the software of the National Center for Advancing Translational Sciences (NIH, UK). Multivariate logistic regression analysis was used to identify relationships between variables. Mean pain level during root canal treatment was 2.2 ± 2.1. The mean anxiety S-DAI score was 27.2 ± 12.5. Fifty percent of men had mild anxiety levels, while in 70% of women anxiety was moderate or high (P = 0.017). Anxiety correlated positively with intraoperative pain (R = 0.406). Multivariate logistic regression analysis revealed that anxiety was significantly associated with intraoperative pain felt by patients (OR = 4.0; 95% C.I. = 1.7-9.3; P = 0.001). Anxious patients were more than twice as likely to feel moderate or intense intraoperative pain during root canal treatment. To know the patient´s degree of anxiety could help the dentist to decide whether to use anxiolytic premedication and/or supplemental local anaesthesia to assure better control of pain during root canal treatment.


Membrane potential depolarization as a triggering mechanism for Vpu-mediated HIV-1 release.
Vpu, a component unique to HIV-1, greatly enhances the efficiency of viral particle release by unclear mechanisms. This Vpu function is intrinsically linked to its channel-like structure, which enables it to interfere with homologous transmembrane structures in infected cells. Because Vpu interacts destructively with host background K(+) channels that set the cell resting potential, we hypothesized that Vpu might trigger viral release by destabilizing the electric field across a budding membrane. Here, we found that the efficiency of Vpu-mediated viral release is inversely correlated with membrane potential polarization. By inhibiting the background K(+) currents, Vpu dissipates the voltage constraint on viral particle discharge. As a proof of concept, we show that HIV-1 release can be accelerated by externally imposed depolarization alone. Our findings identify the trigger of Vpu-mediated release as a manifestation of the general principle of depolarization-stimulated exocytosis.


Endurance training alters alanine and glutamine release from muscle during contractions.
The release of alanine and glutamine from perfused muscle of trained and control animals was investigated. Release rates did not differ between trained and control muscle at rest. During contractions in trained muscle, alanine release was higher than at rest, while glutamine release was transiently increased. Phenylalanine release did not differ between trained and control muscle, implying that protein degradation was not increased in trained muscle. The muscle cellular adaptations to training include a selective modification of amino acid output, which could potentially influence gluconeogenic flux and alter muscle ammonia levels during contractions.


Profound resolution of early atherosclerosis with conjugated linoleic acid.
Conjugated linoleic acid (CLA) refers to a group of positional and geometric isomers of linoleic acid and has been shown to suppress the development of atherosclerosis in experimental models. However, the mechanism involved is unclear although it is believed it may act as a cyclooxygenase inhibitor or as an agonist of the nuclear receptors, peroxisome proliferator activated receptors (PPARs). In this study, we examined the effect of cis-9,trans-11:trans-10,cis-12-CLA (80:20 blend) on the regression of pre-established atherosclerosis. ApoE(-/-) mice fed a 1% cholesterol diet were randomized at 8 weeks to continue receiving the diet supplemented with 1% control saturated fat or 1% CLA blend for a further 8 weeks. CLA supplementation did not simply prevent progression but induced almost complete resolution of atherosclerosis. Although CLA inhibited platelet deposition, as detected by staining of platelet glycoprotein alpha11b beta111a, it did not inhibit COX-mediated generation of prostaglandins in this model. However, PPARalpha and PPARgamma expression was increased in the aorta of the CLA-treated animals. This was coincident with decreased macrophage accumulation and decreased expression of the macrophage scavenger receptor CD36 and increased apoptosis in the aorta in vivo. CLA induces the resolution of atherosclerosis by negatively regulating the expression of pro-inflammatory genes and inducing apoptosis in the atherosclerotic lesion.


Placebo-controlled comparison of candoxatril, an orally active neutral endopeptidase inhibitor, and captopril in patients with chronic heart failure.
To compare the effects on exercise capacity of the neutral endopeptidase inhibitor candoxatril, and the angiotensin converting enzyme inhibitor captopril, in patients with mild to moderate heart failure. In this multi-centre double-blind placebo controlled study, 60 patients with NYHA Class I-III heart failure were randomised to candoxatril 200 mg b.d. (n = 22), captopril 25-50 mg b.d. (n = 23) or placebo (n = 15). Treadmill exercise tests were carried out weekly during a 5-week single-blind placebo run-in phase until a stable baseline was achieved, and repeated at 4 weekly intervals during the 12-week double-blind treatment phase. Nine patients withdrew from the study--four candoxatril and five captopril. The placebo-adjusted increase in exercise duration after 12 weeks was 56 s (95% CI, -26 to +137 s; P = 0.12) with candoxatril and 37 s (-43 to + 117 s; P = 0.29) with captopril. Both candoxatril and captopril were well tolerated and treadmill exercise duration appeared to increase during 12 weeks of therapy but this did not achieve statistical significance. This study tentatively suggests that in patients with heart failure, neutral endopeptidase inhibition may provide similar symptomatic benefits to angiotensin converting enzyme inhibition.


Hypoxia-induced plasticity in the metabolic response of a widespread cichlid.
The African cichlid, Pseudocrenilabrus multicolor victoriae is a eurytopic fish that exhibits high levels of developmental plasticity in response to dissolved oxygen availability. In this study, F1 offspring from three sites in the Mpanga River drainage of Western Uganda characterized by different dissolved oxygen (D.O.) regimes were reared under normoxic or hypoxic conditions. After 1 year, enzymes were measured to determine the tissue metabolic capacity of four different tissues: muscle, heart, brain and liver. The enzymes measured were pyruvate kinase [PK], lactate dehydrogenase [LDH], citrate synthase [CS], and cytochrome C oxidase [CCO], and an additional two, malate dehydrogenase (MDH) and fructose 1,6-bisphosphatase (FBPase), were examined in the liver only. Individuals reared under hypoxia exhibited elevated levels of LDH and CCO in the heart; and depressed activity levels of brain CS and liver CCO and MDH relative to normoxia-reared sibs. Results from this study demonstrate that long-term exposure to hypoxia during development can induce changes in the metabolic capacities of P. multicolor. This flexibility may be important in facilitating persistence in variable and/or novel environments, and in the face of increasing global hypoxia.


Experience-dependent structural plasticity in the adult human brain.
Contrary to assumptions that changes in brain networks are possible only during crucial periods of development, research in the past decade has supported the idea of a permanently plastic brain. Novel experience, altered afferent input due to environmental changes and learning new skills are now recognized as modulators of brain function and underlying neuroanatomic circuitry. Given findings in experiments with animals and the recent discovery of increases in gray and white matter in the adult human brain as a result of learning, the old concept of cognitive reserve, that is the ability to reinforce brain volume in crucial areas and thus provide a greater threshold for age-dependent deficits, has been reinforced. The challenge we face is to unravel the exact nature of the dynamic structural alterations and, ultimately, to be able to use this knowledge for disease management. Understanding normative changes in brain structure that occur as a result of environmental changes and demands is pivotal to understanding the characteristic ability of the brain to adapt.


Changes in Intranetwork Functional Connectivity of Resting State Networks Between Sessions Under Anesthesia in Neurosurgical Patients.
In this study, we evaluated the changes in resting-state networks (RSNs) under anesthesia in neurosurgical patients. RSNs were analyzed in 12 patients with pituitary adenoma presented by chiasma compression operated via standard transsphenoidal approach under propofol anesthesia before and after tumor resection. All the patients had suprasellar tumor extension with compression of the optic chiasma. We investigated second-level effects by contrasting dummy-encoded covariates representing the effects of the sessions (first vs. second) on RSNs. We corrected for multiple comparisons using a false discovery rate of 0.05 (2-sided). Connectivity between the right and left precentral gyri (motor network) decreased significantly from the first to the second session (P = 0.0002), as did the connectivity between the postcentral gyri (P = 0.009). The same was valid for connectivity between the visual cortices (P = 0.0002). The salience network showed a significant decrease in the connectivity of the anterior part of the cingulate gyrus and insular cortex (P = 0.0001). The default mode network showed a decrease in the connectivity between the posterior part of the cingulate gyrus, parietal, and frontal cortices (P = 0.0002). There was no significant correlation between the reduction in connectivity and dose or duration of anesthesia. Different RSNs could be identified under anesthesia and used for intraoperative brain mapping and remapping during tumor resection. However, RSNs showed a significant decrease in connectivity with the continuation of anesthesia.


In vitro antifungal activity of Shikonin against Candida albicans by inducing cellular apoptosis and necrosis.
Our previous studies showed that Shikonin (SK) had a strong anti-Candida albican (C. albicans) activity, especially against some fluconazole-resistant strains, which is probably due to the oxidative damage of SK to C. albicans. In this study, we expanded the antifungal spectrum and evaluate the toxicity of SK. The results indicated that SK also exhibited potent invitro antifungal activities against other pathogenic fungi such as other Candida, Aspergillus, Cryptococcus, and Dermatophytes, but did not display apparent toxicity to the mammalian cells, suggesting that SK is safe to be a potential antifungal drug. Furtherly, we analyze the exact mechanism of SK against C. albicans. We found that SK could induce a series of apoptosis characteristics, including phosphatidylserine externalization, chromatin condensation and fragmentation, decreased cytochrome c oxidase activity as well as caspase activation. In summary, this study highlighted the antifungal activity and mechanism of SK against C. albicans, providing a potential therapeutic strategy for C. albicans infection.


Subcutaneous emphysema.
Subcutaneous emphysema is most often associated with thoracic pathology. The case is presented of a middle-aged woman with shock, abdominal distention and rigidity, and subcutaneous emphysema which resulted from a spontaneous gastric rupture. Subcutaneous emphysema has a variety of origins, including infections and rents in the respiratory tract, gastrointestinal tract, and the skin.


Central nervous system control mechanisms of swallowing: a neuropharmacological perspective.
Neuropharmacological in vivo and in vitro investigations are beginning to provide insight into chemical signaling processes within brainstem networks controlling the individual stages of swallowing. Different subtypes of excitatory amino acid (EAA) receptors operate at the level of solitarial interneurons programming the buccopharyngeal and esophageal stage, as well as motoneurons innervating esophageal striated musculature. Muscarinic cholinoceptors (MAChRs), probably activated via a propriobulbar input, are critically involved in generating output from solitarial neurons to esophageal motoneurons. Inhibition to tonically active GABAA-receptor mediated afferents to solitarial premotor neurons results in rhythmic deglutitive output, reflecting disinhibition of EAA and MACh receptor activity. Motoneuronal EAA receptors may be regulated by a somatostatinergic input arising from solitarial premotoneurons. The available evidence is consistent with a transmitter heterogeneity in esophageal premotor neurons that may operate to provide chemical coding of afferents to the motor output stage of the pattern generator for esophageal peristalsis.


Competence of medical students and residents in psychiatry regarding spirituality, at a South African school of clinical medicine.
Elsewhere, curricula for undergraduate and postgraduate psychiatry have been extended to include the role of spirituality. It has also become important to establish how, within accepted professional boundaries, spirituality should be incorporated appropriately into the current model for South African practice and training. The objectives included exploring, analyzing, and describing the views and experience of local academic psychiatrists on this topic. This study was designed as an explorative, descriptive, phenomenological qualitative investigation. In-depth, semi-structured interviews were conducted with individual academic specialist psychiatrists as the primary data source. Data saturation was achieved after 13 interviews. "Training of spirituality in psychiatry" emerged as one of six main themes from the integrated interview and literature content. All participants proposed that spirituality must be included in undergraduate medical and specialist psychiatric training. They have suggested that a mentorship model should be considered, which implies the reorientation of the teachers of psychiatry in this regard. This view concurred with the international medical literature, recommending that spirituality has to be incorporated into specialist psychiatric practice and training, but within professional boundaries and with all faith traditions and belief systems considered equally.


Rapid reversal of angiotensin converting enzyme inhibition by lisinopril in the perfused rabbit lung.
Lisinopril is a potent competitive inhibitor of purified rabbit lung ACE (dissociation t1/2 = 105 min). To examine reversibility of binding and ACE functional activity in situ, the single-pass extraction (E) of an 125I-lisinopril analogue (351A) and the hydrolysis of an ACE substrate, benz-phe-ala-pro (BPAP) were studied. Lungs were perfused at 50 ml/min with a Krebs-albumin (3%) solution. A bolus containing [14C]dextran, [3H]BPAP, and 351A was injected and (E)351A measured by multiple indicator dilution technique. BPAP metabolism (M) was reflected by the appearance of its hydrolysis product [3H]benz-phe in lung effluent. Control (E)351A was 66 +/- 5% (mean +/- SD, n = 6) and (M)BPAP was 69 +/- 9% (n = 6). Unlabeled lisinopril (30 nmol) in the bolus significantly reduced E(351A) and M(BPAP) to 16 +/- 16% and 3 +/- 3%, respectively. Ten minutes later E(351A) and M(BPAP) had returned to control values. Reduction of E(351A) was partially reversible and M(BPAP) completely reversible after 1 min. After recirculation with 0.25 mM lisinopril for 30 min, however, significant depression of E(351A) was evident for 60 min after exposure to lisinopril was discontinued. Thus, rapid as well as slowly reversible components of inhibition of ACE inhibitor binding can be demonstrated in the perfused rabbit lung.


Asymmetric synthesis of bicyclo[4.3.1]decadienes and bicyclo[3.3.2]decadienes via [6 + 3] trimethylenemethane cycloaddition with tropones.
The cyanosubstituted trimethylenemethane donor undergoes palladium-catalyzed [6 + 3] cycloaddition with a variety of tropones to yield bicyclo[4.3.1]decadienes in excellent regio-, diastereo-, and enantioselectivity. Products of the Pd-TMM [6 + 3] cycloaddition participate in a thermal [3,3] sigmatropic rearrangement to yield bicyclo[3.3.2]decadienes in good yield.


Trapezius muscle rest time during standardised computer work--a comparison of female computer users with and without self-reported neck/shoulder complaints.
Work related musculoskeletal disorders (WMSDs) in the shoulder/neck area are a common and increasing problem among European computer workers, especially women. Long-term low-level workloads with low degree of muscle rest are a potential risk factor for developing WMSDs. The purpose of the present study of female computer users (age 45-65 years) in Denmark and Sweden was to investigate if subjects with self-reported neck/shoulder complaints (cases, N=35) show less trapezius muscle relative rest time (RRT) than controls (N=44) when performing standardised short-term computer work tasks in controlled laboratory conditions. Surface electromyography (EMG) signals were recorded bilaterally from the upper trapezius muscles during a type, edit, precision and colour word stress task. Besides RRT, 10th percentile RMS values were calculated. On the average, 15 of the cases and 18 of the controls showed analysable EMG files per task. For the colour word stress task, the results showed lower RRT values and higher 10th percentile RMS amplitude levels among cases compared to controls. No such signs could be found for the other tasks performed. The present results indicate an increased motor response to a psychological stressor among subjects with self-reported neck/shoulder complaints.


A longitudinal study to explain strategies to change weight and muscles among normal weight and overweight children.
Previous research has indicated that both boys and girls strive for a slim body, with boys having an additional focus on a muscular body build. The current study was designed to evaluate the utility of a biopsychosocial model to explain body image and body change strategies among children. The study evaluated changes over time in body image and strategies to lose weight and increase muscles among 132 normal weight and 67 overweight boys (mean age = 9.23 years) and 158 normal weight and 55 overweight girls (mean age = 9.33 years). The predictive role of BMI, positive and negative affect, self-esteem and perceived sociocultural pressures to lose weight or increase muscle on body image and body change strategies over a 16 month period was evaluated. All participants completed the questionnaire on both occasions. The results demonstrated that both overweight boys and girls were more likely to be dissatisfied with their weight, place more importance on their weight, engage in more strategies to lose weight as well as perceive more pressure to lose weight. Overweight boys and girls were also more likely to report lower levels of self-esteem and positive affect, and higher levels of negative affect, and reported a reduction in their self-esteem over time. Regression analyses demonstrated that among overweight boys, low self-esteem and high levels of perceived pressure to lose weight predicted weight dissatisfaction; for overweight girls, weight dissatisfaction was also predicted by low levels of self-esteem. The implication of these findings in terms of factors contributing to the adoption of health risk behaviors among children is discussed.


Scalp cooling to prevent alopecia after chemotherapy can be considered safe in patients with breast cancer.
With modern scalp cooling equipment cytotoxic damage of hair root cells can be prevented in half of the patients with cancer at high risk of alopecia. However, traditionally doubt has existed whether scalp cooling might facilitate hiding and disseminating scalp skin metastases and thus decrease survival. We discuss this risk using frequency data on metastases in breast cancer from observational and autopsy studies and the Munich cancer registry. They showed the incidence of scalp skin metastases to be very low and not differ between scalp-cooled (0.04-1%) and non scalp-cooled (0.03-3%) patients with breast cancer and in need of chemotherapy. We found it rather unlikely that the incidence of scalp skin metastases might increase at all after scalp cooling, whereas a very small proportion of patients receiving chemotherapy are at risk to develop metastases at this site. Scalp cooling can thus safely be offered to patients treated with alopecia-inducing chemotherapy.


Ultrasonographic demonstration of the pancreatic duct.
During a 4-year period, 375 patients with clinically suggestive pancreatic disease were examined with gray scale ultrasound. In 14 patients, the pancreatic duct itself was demonstrated. All of these had clinical or laboratory evidence for, and radiologically or surgically proven, pancreatic duct obstruction. Etiology of the obstruction was either tumor, chronic pancreatitis, pancreatic abscess, pseudocyst, or stricture of the duct at the papilla of Vater. Two patients had Crohn's disease of the duodenum.


Chiral recognition by biological macromolecules. Partial resolution of racemic enones by albumin.
A novel approach to the optical resolution of racemic enones has been introduced by using the binding properties of the transport protein albumin, which chemically binds preferentially one antipode of some alpha, beta-unsaturated cyclic enones in a reversible manner. Simple separation of the macromolecules by ultrafiltration leads to partial resolution of racemates.


Renin gene and angiotensin II AT1 receptor gene expression in the kidneys of normal and of two-kidney/one-clip rats.
This study aimed to investigate the inter-relation between the angiotensin II (ANG II) AT1 receptor and renin gene expression in rat kidneys. To this end, renin mRNA levels and mRNA levels for AT1a and AT1b were assayed by RNase protection in the kidneys of normal rats, in animals treated with the AT1 antagonist losartan and in rats bearing 0.2-mm left renal artery clips for 2 days. In normal rats, we found a negative correlation between renin mRNA levels and AT1a receptor mRNA levels. Losartan led to a fourfold increase in renin mRNA levels without changing AT1 receptor mRNA levels. Unilateral renal artery clipping increased renin mRNA levels fourfold in the clipped kidney and suppressed renin mRNA levels in the contralateral kidneys. AT1 receptor mRNA levels were not changed in the contralateral intact kidneys, but were significantly decreased by 15-25% in the clipped kidneys. Renin mRNA levels were inversely correlated to AT1a mRNA levels in the clipped, but not in the contralateral, kidneys. Our findings suggest that the systemic activity of the renin angiotensin system has no regulatory influence on renal AT1 receptor gene expression. Renin mRNA levels in normal and in clipped kidneys appear to be negatively determined by the level of AT1a receptor gene expression. Thus modulation of AT1a receptor gene expression could be a pathway for indirect modulation of renin gene expression by ANG II. This conclusion is in agreement with the observation that AT1 receptor antagonists are powerful stimulators of the renin system.


Neuritogenic and chemical properties of guinea pig anterior and posterior root myelin.
In relapsing experimental allergic encephalomyelitis, recurrent demyelination was found in the anterior roots and dorsal root ganglia with minimal involvement of the posterior roots. To determine whether this is an antigen-related phenomenon, the distribution, type and intensity of the lesions in the proximal PNS of guinea pigs immunized with anterior roots or myelin were compared to those of animals immunized with posterior roots or myelin. Homologous anterior roots were less neuritogenic than posterior roots or posterior root myelin. Thin layer chromatography of myelin samples from anterior and posterior roots, dorsal root ganglia and sciatic nerve revealed the presence of a sulfogalactoglycerolipid, tentatively identified as sulfated galactosylglyceride (SGG) in all but the posterior root myelin samples. Although the PNS lesions of relapsing experimental allergic encephalomyelitis appear to recapitulate the regional distribution of SGG, the reason why its presence in anterior roots myelin renders them less neuritogenic is at present not clear.


Role of miR‑181a‑5p in cancer (Review).
MicroRNAs (miRNAs) are non‑coding RNAs (ncRNAs) that can post‑transcriptionally suppress targeted genes. Dysregulated miRNAs are associated with a variety of diseases. MiR‑181a‑5p is a conserved miRNA with the ability to regulate pathological processes, such as angiogenesis, inflammatory response and obesity. Numerous studies have demonstrated that miR‑181a‑5p exerts regulatory influence on cancer development and progression, acting as an oncomiR or tumor inhibitor in various cancer types by impacting multiple hallmarks of tumor. Generally, miR‑181a‑5p binds to target RNA sequences with partial complementarity, resulting in suppression of the targeted genes of miR‑181a‑5p. However, the precise role of miR‑181a‑5p in cancer remains incompletely understood. The present review aims to provide a comprehensive summary of recent research on miR‑181a‑5p, focusing on its involvement in different types of cancer and its potential as a diagnostic and prognostic biomarker, as well as its function in chemoresistance.


Early-life exposure to air pollution associated with food allergy in children: Implications for 'one allergy' concept.
The rapid increase of food allergy (FA) has become the "second wave" of allergy epidemic and is now a major global public health concern. Mounting evidence indicates that early life exposure to air pollution is associated with the "first wave" of allergy epidemic (including asthma, allergic rhinitis and eczema) in children, but little is known about its association with FA. We hypothesize FA has triple exposure pathways, gut-skin-airway, and investigate the effects of airway exposure to outdoor and indoor air pollution on childhood FA. A cohort study of 2598 preschool children aged 3-6 years old was conducted in Changsha, China. The prevalence of FA was surveyed using a standard questionnaire by International Study of Asthma and Allergies in Childhood (ISAAC). Exposure to indoor air pollution was assessed by four indicators: new furniture, redecoration, mold or dampness, and window condensation. Exposure to outdoor air pollution was evaluated by the concentrations of PM10, SO2 and NO2, which were obtained from the monitored stations. Both prenatal and postnatal exposure windows were considered. The association between exposure to outdoor/indoor air pollution and childhood FA was estimated by multiple logistic regression models using odds ratio (OR) and a 95% confidence interval (CI). A total of 14.9% children reported FA. The prevalence was significantly associated with exposure to indoor air pollution, OR (95% CI) = 1.93 (1.35-2.75) for prenatal exposure to mold/dampness and 1.49 (1.07-2.10) and 1.41 (1.04-1.89) respectively for postnatal exposure to new furniture and window condensation. The prevalence of FA was also associated with prenatal and postnatal exposure to outdoor air pollution, particularly the traffic-related air pollutant NO2, with adjusted ORs (95% Cls) respectively 1.24 (1.00-1.54) and 1.38 (1.03-1.85) per interquartile range (IQR) increase. Sensitivity analysis showed that the association between outdoor/indoor air pollution and childhood FA was significant only in young children aged 3-4 years. Early-life exposure to high levels of outdoor and indoor air pollution in China due to the rapid economic growth and fast urbanization in the past decades may contribute to the rapid increase of food allergy (FA) in children. Our study indicates that, in addition to gut and skin, airway may be a new route of food sensitization. Air pollution leads to the first and second waves of allergy epidemics, suggesting a concept of 'one allergy' disease.


The neurological insights of the emerging coronaviruses.
Emerging Viral diseases are incredibly infectious and proficient in inducing pandemics. Unlike the previous emerging coronaviruses (ECoVs) which neurological complexities were uncommon, with neurological features exhibition at 14-25 days post-onset, yet with critical outcomes exhibiting >50% mortality in central nervous (CNS) presenting pathologies. The COVID 19 neurological consequences occur more frequently even in mild cases, presenting with CNS involvement in up to 25%, musculoskeletal and peripheral manifestation (PNM). Through preceding ECoVs case reports, the PNM not linked to fatal outcomes, however, required, repeated neuro-imaging as notable CT and MRI changes appeared as late as 21 days while the likelihood of Cerebrospinal fluid to test positive for ECoV was 25%, only in the CNS presenting cases. Owing to 44-60% myalgia presentation, risk of the high inflammatory state, and coagulation cascade abnormalities reported in ECoVs, testing for C-reactive protein, serum creatine kinase, and D-dimer level is mandatory. Presently, there is no antiviral medication or vaccination for the ECoVs, early induction of antiviral drugs remains the backbone of management. Neurologically, the therapeutic dosages of anticoagulants are linked to the high incidence of thrombotic complexities, while methylprednisolone is associated with myopathy. Future studies expected to apply more neuro-imaging techniques for CNS exploration and further explore the pathogenesis of the COVID 19 myalgia, anosmia/ageusia reported in the majority of the initial cases.


The ozone sensitivity of five poplar clones is not related to stomatal conductance, constitutive antioxidant levels and morphology of leaves.
Ground-level ozone (O3) is an important phytotoxic air pollutant in China. In order to compare the sensitivity of common poplar clones to O3 in China and explore the possible mechanism, five poplar clones, clone DQ (Populus cathayana), clone 84 K (P. alba × P. glandulosa), clone WQ156 (P. deltoids × P. cathayana), clone 546 (P. deltoides cv. '55/56' × P. deltoides cv. 'Imperial') and clone 107 (P. euramericana cv. '74/76') were exposed to four O3 treatments. According to the date of the initial visible O3 symptom and the slopes of O3 exposure-response relationships with the relative light-saturated rate of CO2 assimilation, we found that clone DQ and clone 546 were the most sensitive to O3, clone 84 K and clone WQ156 were the less sensitive, and clone 107 was the most tolerant, which could provide a basis to select O3 tolerant clones for poplar planting at areas with serious O3 pollution. Elevated O3 significantly reduced photosynthetic parameters, total phenols content, potential antioxidant capacity, leaf mass per area and biomass of five poplar clones, and there were significant interactions between O3 and clones for most photosynthetic parameters. Elevated O3 also significantly increased malondialdehyde content and total ascorbate content. The responses of total antioxidant capacity for poplar clones to elevated O3 were different, as indicated by the increase for clone 107 and reduction for other clones under elevated O3 treatment. Our results on the sensitivity of different poplar clones to O3 are not related to leaf stomatal conductance, leaf constitutive antioxidant levels or leaf morphology of plant grown in clean air. The possible reason is little difference in leaf traits among clones within close species, suggesting that more properties of plants should be considered for exploring the sensitivity mechanism of close species, such as mesophyll conductance, antioxidant enzyme activity and apoplastic antioxidants.


Structural and biochemical characterization of the pleckstrin homology domain of the RhoGEF P-Rex2 and its regulation by PIP3.
P-Rex family Rho guanine-nucleotide exchange factors are important regulators of cell motility through their activation of a subset of small GTPases. Both P-Rex1 and P-Rex2 have also been implicated in the progression of certain cancers, including breast cancer and melanoma. Although these molecules display a high level of homology, differences exist in tissue distribution, physiological function, and regulation at the molecular level. Here, we sought to compare the P-Rex2 pleckstrin homology (PH) domain structure and ability to interact with PIP3 with those of P-Rex1. The 1.9 Å crystal structure of the P-Rex2 PH domain reveals conformational differences in the loop regions, yet biochemical studies indicate that the interaction of the P-Rex2 PH domain with PIP3 is very similar to that of P-Rex1. Binding of the PH domain to PIP3 is critical for P-Rex2 activity but not membrane localization, as previously demonstrated for P-Rex1. These studies serve as a starting point in the identification of P-Rex structural features that are divergent between isoforms and could be exploited for the design of P-Rex selective compounds.


A comparative analysis of antidepressant and anti-suicidal effects of repeated ketamine infusions in elderly and younger adults with depression.
This study aims to investigate the differences in safety and antidepressant effects of multi-infusion ketamine treatment between elderly and young adults with depression. The safety, antidepressant, and anti-suicidal effects of multi-infusion ketamine were compared between 19 elderly (≥50 years) and 116 younger (<50 years) adults with depression; all were treated with six ketamine infusions (0.5 mg/kg). Montgomery-Åsberg Depression Rating Scale (MADRS) was used to measure the depressive symptoms, and suicidal ideation was measured with Beck Scale for Suicide Ideation (SSI)-part 1, Hamilton Rating Scale for Depression (HAMD) item 3, and (MADRS) item 10. Dissociative and psychotomimetic symptoms were evaluated based on the Clinician-Administered Dissociative States Scale (CADSS) and the Brief Psychiatric Rating Scale (BPRS)-four items. Multi-Ketamine infusions resulted in a lower (trend) antidepressant response (37.1 % versus 57.8 %) and antidepressant remission (15.8 % versus 47.4 %) in elderly patients with depression compared with younger patients with depression (all ps > 0.05). Interestingly, elderly patients with depression had a higher MADRS score after six ketamine infusions compared with younger patients (p = 0.04). No significant differences in SSI-part 1 scores, HAMD item 3 scores, MADRS item 10 scores, CADSS scores, and BPRS-four items scores were found between the two groups at any assessment point (all ps > 0.05). Our study shows that repeated-dose infusions of ketamine may be a feasible treatment strategy in elderly Chinese patients with depression; however, elderly patients with depression may be less responsive to ketamine compared with younger adults with depression.


The effects of thiamin on the tissue distribution of lead.
The effects of thiamin on the tissue distribution of lead were evaluated in Sprague-Dawley rats exposed to 1000 ppm lead acetate in drinking water and treated daily with thiamin (25 or 50 mg kg-1 body weight, i.p.), calcium ethylenediamine tetraacetic acid (50 mg kg-1 body weight, i.p.) or their combination for 8 weeks. The subtoxic dose of lead did not alter weight gains, feed and water consumption during the treatment period. Thiamin decreased the blood (P less than 0.0001), liver (P less than 0.0001) and kidney (P less than 0.0001) concentrations of lead. Thiamin (50 mg kg-1 body weight) reduced the lead concentrations in tissues more effectively than thiamin (25 mg kg-1 body weight). The combined treatment was more effective than the respective individual treatments.


Surface antigen expression on bladder tumor cells induced by bacillus Calmette-Guérin (BCG): A role of BCG internalization into tumor cells.
The antitumor mechanisms of bacillus Calmette-Guérin (BCG) against bladder cancer is still unclear. We previously reported that BCG was internalized by and survived within murine bladder tumor cells (MBT-2) for at least 40 days. In the present study, we investigated the effect of BCG on the surface antigen expression of bladder tumor cells and the characteristics of these cells as antigen-presenting cells in vitro. Surface antigen (major histocompatibility complex (MHC) Class II, CD1, CD80 and intercellular adhesion molecule-1 (ICAM-1)) expression on BCG-treated murine (MBT-2) and human (T-24, J82) bladder tumor cells were analyzed using flow cytometry. The production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) from murine lymphocytes sensitized with BCG or BCG-treated tumor cells were also investigated. The expressions of MHC Class II, CD1, CD80 and ICAM-1 were augmented in all of the bladder tumor cell lines used; however, they were augmented to varying degrees among the cell lines that were treated with live BCG. Heat-killed BCG had little or no effect. When murine lymph node cells sensitized with BCG or BCG-treated MBT-2 cells were cocultured with BCG-treated MBT-2 cells, significant amounts of IL-2 and IFN-gamma were produced in the culture medium. BCG induced the augmented expression of surface antigens, such as MHC Class II, CD1, CD80 and ICAM-1, of bladder tumor cells. Furthermore, BCG-treated MBT-2 cells could stimulate BCG-sensitized lymphocytes to produce IL-2 and IFN-gamma. These results strongly suggest that bladder tumor cells gained the characteristics and functions of antigen-presenting cells (APC).


Axonal transport and the delivery of pre-synaptic components.
The mechanisms for delivering components to nerve terminals are diverse and highly regulated. The diversity of kinesin motors alone is insufficient to account for the specificity of delivery. Additional specificity and control are contributed by adaptor proteins and associated regulatory molecules. The interaction of cargos with these complexes can confer distinct behaviors on the transport of synaptic organelles. The rich regulatory mechanisms of transport that are only now emerging as the cargo-motor complexes are defined and subsequent local events that regulate their dynamic relationship are examined. Here we review recent studies of kinesin-related axonal transport of three crucial synaptic components, Piccolo-bassoon Transport Vesicles (PTVs), Synaptic Vesicle Precursors (SVPs), and mitochondria, and the mechanisms that modulate their transport.


miRNA-21-3p targeting of FGF2 suppresses autophagy of bovine ovarian granulosa cells through AKT/mTOR pathway.
It is widely thought that the main reason for ovarian follicular atresia is apoptosis of granulosa cells, however, accumulating evidence suggests that autophagy plays a role in the fate of granulosa cells. Although epigenetic regulation including miR-21-3p associated with autophagy process has been reported in many cancer types, nevertheless, the mechanism of miR-21-3p in bovine ovary is poorly understood. In the present study, bovine ovarian granulosa cells (BGCs) were used as a model to elucidate the autophagy and role of miR-21-3p in a cattle ovary. The results from gene expression and tagged autophagosomes showed the autophagy in BGCs and miR-21-3p was identified as an important miRNA regulating autophagy of BGCs. The current results indicated that FGF2 was a validated target of miR-21-3p in autophagy regulation of BGCs according to the results from FGF2 luciferase reporter assays and FGF2 overexpression (oe-FGF2) or small interference (si-FGF2). Transfection of miR-21-3p mimic and si-FGF2 plasmids resulted in decreasing phosphorylated AKT and mTOR, while transfection of miR-21-3p inhibitor and oe-FGF2 increased the phosphorylated level of AKT and mTOR in BGCs. These data indicate that regulation of miR-21-3p on BGCs autophagy through AKT/mTOR pathway. In summary, this study suggests that miR-21-3p targets FGF2 to inhibit BGCs autophagy by repressing AKT/mTOR signaling.


Structure-activity relationships for mini atrial natriuretic peptide by proline-scanning mutagenesis and shortening of peptide backbone.
MiniANP is a synthetic pentadecapeptide analogue of atrial natriuretic polypeptide (ANP). We have used the proline-scanning mutagenesis and the analogue peptides with shorter backbones to characterize the turn-like conformation at residue 6-9 and an extended structure of Gly5-Gly6 as the receptor-bound structure of miniANP. A docking study of miniANP at the binding site of the type A natriuretic peptide receptor (NPR-A) supported the deduced conformation in the receptor-bound structure.


Electrical stimulation of the medial prefrontal cortex supports both 'pure reward' and 'reward-escape' behavior in rats.
In female Sprague-Dawley rats, 8 of 12 medial prefrontal cortex (MPFC) sites that yielded criterion self-stimulation behavior supported only self-stimulation, i.e. were 'pure reward' in type. The remaining 4 sites supported behavior to escape from experimenter-administered stimulation of the same parameters as well, i.e. were 'reward-escape' in type. 'Pure reward' and 'reward-escape' sites in the MPFC were distinguished by both the magnitude and temporal form of the escape response functions generated, and by the prevalence of 'pounce-back', a vigorous and repetitive barpressing during the 3-s MPFC stimulation-escape interval produced by an effective barpress. The finding that both 'pure reward' and 'reward-escape' patterns of behavior can be elicited by stimulation of the MPFC provides a basis for further assessment of similarities and differences in medial prefrontal cortical and lateral hypothalamic (LH) 'reward' systems. It is suggested that 'reward-escape' in the MPFC may be mediated by the activity of 'reward' neurons which respond to stimulus offset, rather than by a secondary aversive process as is proposed to underlie 'reward-escape' in the LH.


CMV-specific central memory T cells reside in bone marrow.
CMV-specific CD8(+) T cell responses in peripheral blood (PB) are characterized by a preponderance of effector and effector memory T cells. CMV-specific central memory T cells (T(CM)), which are considered crucial in maintaining long-term immunity, are rarely detectable in PB. In this study we have analyzed differentiation and function of CMV pp65-specific CD8(+) T cells in paired samples of human PB and BM using intracellular cytokine and tetramer staining. Overall frequencies of CMV pp65-specific T cells were similar in PB compared to BM; however, CMV-specific CD45RA(-)CCR7(+) T(CM) were almost exclusively detectable in BM, which was not related to a general accumulation of T(CM) in BM. In vitro, CMV-specific T cells could be more efficiently expanded from BM (median 128-fold, n=6) than from PB (median 72-fold, p=0.01). Taken together, these data show that the BM is a compartment harboring CMV-specific T(CM) and underline the concept of the BM as a secondary immune organ. CMV specific BM-derived T(CM) might be a valuable source for generating T cells for adoptive transfer.


Comparison of serological methods with PCR-based methods for the diagnosis of community-acquired pneumonia caused by atypical bacteria.
The diagnosis of community-acquired pneumonia (CAP) caused by Legionella pneumophila, Mycoplasma pneumoniae, and Chlamydophila pneumoniae is traditionally based on cultures and serology, which have special requirements, are time-consuming, and offer delayed results that limit their clinical usefulness of these techniques. We sought to develop a multiplex PCR (mPCR) method to diagnosis these bacterial infections in CAP patients and to compare the diagnostic yields obtained from mPCR of nasopharyngeal aspirates (NPAs), nasopharyngeal swabs (NPSs), and induced sputum (IS) with those obtained with specific PCR commercial kits, paired serology, and urinary antigen. A total of 225 persons were included. Of these, 10 patients showed serological evidence of L. pneumophila infection, 30 of M. pneumoniae, and 18 of C. pneumoniae; 20 individuals showed no CAP. The sensitivities were mPCR-NPS = 23.1%, mPCR-IS = 57.1%, Seeplex®-IS = 52.4%, and Speed-oligo®-NPA/NPS = 11.1%, and the specificities were mPCR-NPS = 97.1%, mPCR-IS = 77.8%, Seeplex®-IS = 92.6%, and Speed-oligo®-NPA/NPS = 96.1%. The concordance between tests was poor (kappa <0.4), except for the concordance between mPCR and the commercial kit in IS (0.67). In individuals with no evidence of CAP, positive reactions were observed in paired serology and in all PCRs. All PCRs had good specificity but low sensitivity in nasopharyngeal samples. The sensitivity of mPCR and Seeplex® in IS was approximately 60%; thus, better diagnostic techniques for these three bacteria are required.


Age and sex related variations of adult human ocular volumes in the Sri Lankan population: an evaluation using magnetic resonance imaging.
Variations in the human ocular volumes are related to ocular pathologies including congenital glaucoma, microphthalmus, buphthalmus, and macrophthalmus. As the currently published reference ocular volumes are prone to physiological and racial variations, population specific values may provide more precision in ophthalmological interventions. This study was conducted to assess the age and sex dependent differences in ocular volumes in Sri Lankan individuals using magnetic resonance imaging (MRI). The study was undertaken using the brain MRI scans from 200 patients which were reported as normal. Study sample consisted of patients between 18 years and 90 years of age with 91 male subjects and 109 female subjects. Two independent observers measured ocular volumes using a software-based method and an MRI planimetry based method. Age and sex of the study participants were recorded for the further analysis. Statistically significant differences in both ocular volumes were found between males and females (p < 0.05) when using both volume analysis methods. The mean ± SD ocular volumes obtained as right software based volume, right MRI planimetry volume, left software based volume and left MRI planimetry volume were 6.8 ± 0.6, 6.0 ± 0.6, 6.6 ± 0.7 and 5.9 ± 0.6 cm3 in females and 6.9 ± 0.8, 6.3 ± 0.7, 6.9 ± 0.8 and 6.2 ± 0.7 cm3 in males. While software-based measurements show a significant linear correlation with age in both eyeball volumes, MRI planimetry measurement showed a significant linear correlation with age only in the left eyeball (p < 0.05). Weak negative correlations were found with age in right ocular volume in both MRI planimetry based (r = -0.121) and software based (r = -0.168) measurements and in left ocular volume in MRI planimetry based (r = -0.151) and software based (r = -0.179) measurements. Furthermore, ocular volumes obtained from the software-based method were significantly greater than the MRI planimetry based ocular volumes (p < 0.05) in both eyes, despite having a strong positive correlation. The mean ocular volumes obtained from this study revealed a significant variation between the right and left eyes as well as a sexual dimorphism. Moreover, since the two measurement methods show a significant difference, the choice of measurement method should depend on the required accuracy of the eye volume decided with respect to the clinical implication. Since there are no reference values for Sri Lankan adult ocular volumes, this study may serve that purpose in the current population, while supporting ophthalmologists and radiologists to quantitatively evaluate ocular pathologies and to follow precise interventions.


[Establish efficiently transformation system of the Isatis indigotica].
To build up a method of efficiently transforming Isatis indigotica with the Agrobacterium. Two kinds of Agrobocterium: ATCC 15834 and RI1601 were used to treat different fraction of Isatis indigotica. Explored the effect of cocultured, different antibiotic concentration and the media on multiply the hairy roots. The explants with out coculture had had higher survival rate rooting rate and earlier sprout time. After ultrasonic treatment of plant, the indacement rate was two times than that of untreated one; The best antibiotic concentration was 400 mg/L; The proportion of the grow speed of the hairy root in the liquid culture media was 2 - 3 times than that of solid culture media, and 37 times of common roots. The method is useful for establishing an efficiently transformatiem system of Isatis indigotica by Agrobacterium.


Family-based association of a low producing lymphotoxin-alpha allele with reduced Plasmodium falciparum parasitemia.
Tumor necrosis factor (TNF)-related genes are thought to play a role in human malaria. TNF polymorphisms have been associated with severe malaria, mild malaria, and parasitemia. Lymphotoxin-alpha gene (LTA) that belongs to the TNF family is one such candidate gene. Here we report the family-based association analysis of a cis-regulatory lymphotoxin-alpha polymorphism with parasitemia in two independent populations living in Burkina Faso. Analysis of 199 subjects (34 families) living in an urban endemic area revealed the association of the low producing LTA+80A allele with reduced parasitemia. Furthermore, there was evidence of significant LTA+80-by-age and LTA+80-by-gender interactions. In another set of 318 residents (55 families) in a rural endemic area, we found both the association of the low producing LTA+80A allele with reduced parasitemia and LTA+80-by-age and LTA+80-by-gender interactions. This study suggests that LTA+80 polymorphism influences parasitemia and acts in an age- and gender-dependent manner.