Electro-enhanced solid-phase microextraction with membrane protection for enrichment of bisphenols in canned meat.
Bisphenols (BPs), one of the endocrine disrupting chemicals (EDCs), are harmful to health with the potential carcinogenicity, teratogenicity and neurotoxicity. In this paper, we developed an electro-enhanced solid-phase microextraction with membrane protection method to enrich BPs. The extraction time can be shortened to 10 min with the electric field, and the prepared fiber can be repetitively used 100 times with the membrane protection. The factors affecting extraction such potential, pH, extraction temperature and extraction time were optimized. Under optimal conditions, the linear ranges were 2-600 ng mL-1 for BPAF and 2-800 ng mL-1 for the other four BPs (BPB, BPF, BPA, BPZ), and the limits of detection (S/N=3) were from 0.09 (BPAF) to 0.41 (BPA) ng mL-1, with intra-day and inter-day precisions <5.16% and 9.29%, respectively. The developed method has been applied to the determination of trace BPs in canned meat with the recoveries of 73%-117%, showing a promising application for pretreatment in complex samples.


Endoscopic water jets used to ablate Barrett's esophagus: preliminary results of a new technique.
The optimal management of Barrett's esophagus, a precursor to esophageal adenocarcinoma, remains controversial. Current therapy includes surveillance and ablative or resection techniques of varying safety and efficacy. This study aimed to determine the feasibility of a new catheter-based, endoscopic water jet ablation technique. A high-pressure flexible catheter that can be passed through the working port of a standard gastroscope was used. The catheter had micro-drilled holes on one side near the tip. A 1-cm water jet was delivered under foot pedal control and endoscopic view at pressures adjusted from 150 to 400 psi. After approval from the authors' Institutional Review Board, tissue segments from fresh esophagectomy specimens were ablated by the catheter without use of an endoscope. Using gross appearance and histologic analysis, variable ablation pressures and times were evaluated. Using variable pressures and times, 11 ablation sessions were performed: 5 for normal esophagus, 4 for normal stomach, and 2 across the gastroesophageal junction in the setting of Barrett's esophagus. Ablation pressures of 150 to 300 psi for 30 to 60 s resulted in selective ablation of mucosa with preservation of the submucosa and muscularis propria. The depth of the ablation was determined by gross inspection at the time of ablation and confirmed by histologic evaluation. There was no embedding of epithelial cells in the muscularis propria. In a single normal esophagus specimen, a jet applied at 400 psi for 120 s in a confined area resulted in gross perforation. Selective ablation of esophageal and gastric epithelium using a catheter-based water jet ablation technique is feasible. The preliminary data from this study investigating a nonendoscopic technique show that the mucosa can be removed with preservation of the underlying submucosa and muscular layers. Further studies are warranted that focus on defining more precisely the pressure and duration required for optimal results and the practical application of this technique endoscopically.


Inhibition by Sr2+ of specific mitochondrial Ca2+-efflux pathways.
The effect of Sr2+ on the set point for external Ca2+ was studied in rat heart and liver mitochondria with the aid of a Ca2+-sensitive electrode. In respiring mitochondria the set point is determined by the rates of Ca2+ influx on the Ca2+ uniporter and efflux by various mechanisms. We studied the Ca2+-Na+ exchange pathway in heart mitochondria and the delta psi-modulated efflux pathway in liver mitochondria. Prior accumulation of Sr2+ was found to shift the set points towards lower external Ca2+ both in heart mitochondria under conditions of Ca2+-Na+ exchange and in liver mitochondria under conditions that should promote opening of the delta psi-modulated pathway. The effect on the set point was found to be due to inhibition of Ca2+ efflux by Sr2+ taken up by the mitochondria, while Sr2+ efflux was too slow to be measurable.


Fecal incontinence and associated pelvic floor dysfunction during and one year after the first pregnancy.
Obstetric anal sphincter injury is an important risk factor for postpartum fecal incontinence but few studies have reported fecal incontinence occurring, even during pregnancy. The first objective of this study was to examine the prevalence of fecal incontinence, obstructed defecation and vaginal bulging early and late in pregnancy and postpartum. The second objective was to assess the association between symptoms in pregnancy, delivery characteristics, and bowel and vaginal bulging symptoms at 1 year postpartum. This prospective cohort study was conducted between October 2014 and October 2017, including 898 nulliparous women enrolled with the maternity healthcare service in Örebro County, Sweden. The women responded to questionnaires regarding pelvic floor dysfunction in early and late pregnancy and at 8 weeks and 1 year postpartum. The data were analyzed using random effect logistic models estimating odds ratios (ORs) and generalized linear models estimating relative risks, with 95% confidence intervals (CIs). At 1 year postpartum, the prevalence of fecal incontinence, obstructed defecation and vaginal bulging was 6% (40/694), 28% (197/699) and 8% (56/695), respectively. Among women with vaginal delivery, the risk of fecal incontinence and vaginal bulging increased significantly both in late pregnancy, with ORs of 3.4 (95% CI 1.5-7.7) and 3.6 (95% CI 1.6-8.1), respectively, and at 1 year postpartum, with ORs of 5.0 (95% CI 2.1-11.5) and 8.3 (95% CI 3.8-18.1), respectively, compared with early pregnancy. Among all women, factors associated with increased prevalence of fecal incontinence 1 year postpartum were fecal incontinence during pregnancy (adjusted relative risk [aRR] 7.4; 95% CI 4.1-13.3), obstructed defecation during pregnancy (aRR 2.0; 95% CI 1.1-3.9) and concurrent obstructed defecation (aRR 2.4; 95% CI 1.3-4.5). This prospective study shows an increased risk of fecal incontinence by late pregnancy, suggesting that the pregnancy itself may be involved in the development of postpartum fecal incontinence. Obstructed defecation during pregnancy and postpartum was found to be associated with increased risk of fecal incontinence postpartum, indicating that postpartum fecal incontinence may be a result of incomplete bowel emptying.


Chimeric deletion mutation of rpoC2 underlies the leaf-patterning of Clivia miniata var. variegata.
The deletion mutated rpoC2 leads to yellow stripes of Clivia miniata var. variegata by down regulating the transcription of 28 chloroplast genes and disturbing chloroplast biogenesis and thylakoid membrane development. Clivia miniata var. variegata (Cmvv) is a common mutant of Clivia miniata but its genetic basis is unclear. Here, we found that a 425 bp deletion mutation of chloroplast rpoC2 underlies the yellow stripes (YSs) of Cmvv. Both RNA polymerase PEP and NEP coexist in seed-plant chloroplasts and the β″ subunit of PEP is encoded by rpoC2. The rpoC2 mutation changed the discontinuous cleft domain required to form the PEP central cleft for DNA binding from 1103 to 59 aa. RNA-Seq revealed that 28 chloroplast genes (cpDEGs) were all down-regulated in YSs, of which, four involved in chloroplast protein translation and 21 of photosynthesis system (PS)I, PSII, cytochrome b6/f complex and ATP synthase are crucial for chloroplast biogenesis/development. The accuracy and reliability of RNA-Seq was verified by qRT-PCR. Moreover, the chlorophyll (Chl) a/b content, ratio of Chla/Chlb and photosynthetic rate (Pn) of YS decreased significantly. Meanwhile, chloroplasts of the YS mesophyll cells were smaller, irregular in shape, contain almost no thylakoid membrane, and even proplastid was found in YS. These findings indicate that the rpoC2 mutation down-regulated expression of the 28 cpDEGs, which disturb chloroplast biogenesis and its thylakoid membrane development. Thus, there are not enough PSI and II components to bind Chl, so that the corresponding areas of the leaf are yellow and show a low Pn. In this study, the molecular mechanism of three phenotypes of F1 (Cmvv ♀ × C. miniata ♂) was revealed, which lays a foundation for the breeding of variegated plants.


A PKM2 inhibitor induces apoptosis and autophagy through JAK2 in human oral squamous cell carcinoma cells.
The enzyme pyruvate kinase M2 (PKM2) is involved in glycolysis, which plays an important role in the regulation of tumor progression. In this study, we investigated the anti-tumor activity of N-(4-(3-(3-(methylamino)-3-oxopropyl)-5-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)-1H-pyrazol-1-yl)phenyl)propiolamide (MTP), a PKM2 inhibitor, in oral squamous cell carcinoma (OSCC) cells. Our results showed that MTP inhibited cell growth with IC50 values of 0.59 μM and 0.78 μM in SCC2095 and HSC-3 OSCC cells, respectively. MTP induced caspase-dependent apoptosis, which was associated with the modulation of PKM2 and oncogenic biomarkers epidermal growth factor receptor and β-catenin. In addition, MTP increased the generation of reactive oxygen species (ROS) and modulated the expression of autophagic gene products, including LC3B-II and p62. Western blotting showed that MTP inhibited Janus kinase 2 (JAK2) signaling, and JAK2 overexpression partially reversed MTP-mediated cytotoxicity. Taken together, these data indicate the potential use of MTP as a therapeutic agent for OSCC.


Exploring Approaches to Successfully Achieving Comprehensive Order Set Adoption.
Healthcare providers are challenged to efficiently achieve evidence-based care for patients and their families. Clinical decision support tools are being introduced to facilitate knowledge transfer to care providers. Order sets are one of the tools. Using the realist review methodology and focus group of interview participants, this paper retrospectively explored relationships that lead to successful order set adoption. Findings demonstrate the need for regular review of clinical context. Technology can offer some enhancements but also introduces complex challenges. Ongoing software development is needed to improve delivery formats as well as incorporate effective tools for efficient continuous quality improvement.


Viable spermatozoa in the bladder after electroejaculation of lion-tailed macaques (Macaca silenus).
The bladder of 6 lion-tailed macaques was emptied and flushed with sterile saline. TALP-Hepes buffer was infused and the animals were electroejaculated. After electroejaculation, the semen quality was determined in the ejaculate and the bladder infusate. Of the 15 ejaculates analysed, a mean (+/- s.e.m.) sperm count of 133.8 (+/- 30.7) x 10(6) with 69.5 (+/- 6.0)% motility was obtained in the infusate as compared to the sperm count of 72.4 (+/- 38.6) x 10(6) with significantly lower (47.7 +/- 5.8%) motility in the ejaculate.


Lymph node metastasis and lymphadenectomy for carcinoma in the gastric cardia: clinical experience.
To suggest improved guidelines for treating lymphatic spread and to understand the results of extended lymphadenectomy. A total of 141 cases of cardia carcinoma admitted to our hospital during 1980 to 1993 were analyzed. 123 of 141 cases underwent resection. The incidence of mediastinal lymph node involvement rose from 8.7% in cases with 1-2 cm invasion to 52.9% in cases with more than 4 cm invasion into the esophagus. The incidence of abdominal lymph node involvement also rose with increasing esophageal invasion. Paraaortic lymph node metastasis was frequently observed in cases of more than 1 cm esophageal invasion. D4 lymphadenectomy (extended lymphadenectomy) revealed better results for the cases with lymph node involvement (P<0.05). There was no difference in the incidence of postoperative complication or in operative death, between the abdominal approach and the thoracoabdominal approach, or between the D4 lymphadenectomy and D2 lymphadenectomy. Sufficient mediastinal and abdominal lymphadenectomy under a clear wide surgical field achieved through the thoracoabdominal approach appeared to bring better results.


Spatial distribution and ecological assessment of nickel in sediments of a typical small plateau lake from Yunnan Province, China.
Nickel (Ni) in small plateau lake sediments plays an important role in influencing the quality of lake ecosystems with a high degree of endemism and toxicity. This paper focuses on the spatial distribution and ecological risks of nickel in the sediments of Jianhu Lake, a small plateau lake in China, and the influence of pH and total organic carbon (TOC) on nickel concentrations. The results showed that average total nickel concentrations were 138.99 ± 57.57 mg/kg (n = 38) and 184.31 ± 92.12 mg/kg (n = 60) in surface sediments (0-10 cm top layer) and sediment cores (0-75 cm depth), respectively, and that the residual fraction was the main form of nickel. Simultaneously, through a semivariogram model, strong spatial dependence among pH, TOC, and the oxidizable fraction was revealed, whereas total nickel, exchangeable and the weak acid soluble fraction, reducible fraction, and residual fraction showed moderate spatial dependence. The vertical distribution revealed that nickel accumulated mainly in the bottom 5 cm (70-75 cm) of the sediment layer and that the pH was higher there, whereas TOC was concentrated mainly in the top 5 cm of sediment. Using geoaccumulation and a potential ecological risk index, moderate nickel pollution and moderate risk levels were found in most surface sediments, but moderate nickel pollution and high risk levels were observed in most sediment cores. In addition, pH and TOC were found to have a strong effect on the distribution and concentration of nickel and its fractions in the small plateau lake. In summary, nickel posed a certain degree of pollution and ecological risk, which deserves attention in the sediments of small plateau lakes.


Radiolabeled DNase, a potential indicator for noninvasive detection of tissue damage.
Pancreatic DNase I was labeled with 131I or fluorescamine and injected IV into NMRI mice bearing a sarcoma 180. Of the injected tracer, 1.5%-2% was found to accumulate per g tumor. In sections of tumor tissue DNase was localized in damaged cells in solid and necrotic tumor regions. This binding is most probably due to specific interaction of DNase with actin, an ubiquitous cytoskeletal protein. Two-component blood clearance with a rapid first component (two-thirds of applied radioactivity) was observed. The labeled tumor could easily be visualized by gamma camera imaging. The findings suggest DNase to be a potent radiopharmaceutical for imaging damaged tissue, occurring in malignant tumors as well as in infarcts, and inflammatory lesions.


Chloroquine enhances the number of IL-10 producing cells and the expression of B7-2 and ICAM-1 in in vitro-cultured PBMC.
Chloroquine is prescribed as both an anti-malarial and an anti-inflammatory drug. However, its immuno-modulating effects remain largely unclear. Previous studies have shown that chloroquine inhibits antigen-induced proliferation, implying immuno-suppressive effects. In this study, we examined whether the inhibition of the proliferation reflects changes in the surface molecules that are important for T-cell activation and whether chloroquine affects the balance between pro- and anti-inflammatory cytokines. Chloroquine elevated the expression of the costimulatory and adhesion molecules B7-2 (CD86) and ICAM-1 (CD54) in peripheral mononuclear cells (PBMC). An increased percentage of CD14+ cells was also observed, and within this cell population, an increase in ICAM-1 expression was revealed by double-staining experiments. Assessment of the frequencies of interleukin (IL)-10 and interferon (IFN)-gamma-producing cells in in vitro-cultivated PBMCs showed that the ratio between pro- and anti-inflammatory cytokines changed after exposure to chloroquine, favouring anti-inflammatory immune responses. This effect was mainly because of increased frequencies of IL-10-producing cells and was seen with or without the presence of stimulating antigens or mitogens. Our findings indicate that chloroquine affects the direction of the lymphocyte stimulation towards an anti-inflammatory response by affecting the antigen-presenting cells (APC) and the balance between pro- and anti-inflammatory cytokines, rather than generally inhibiting cytokine production.


Dedifferentiated chondrosarcoma with a rhabdomyosarcomatous component.
A case of dedifferentiated chondrosarcoma arising in the femur with metastasis to the lung is described. On immunoperoxidase studies and electron microscopic examination, both the anaplastic soft tissue extension of this tumor in the hip and the metastatic lung nodule showed evidence of rhabdomyosarcomatous differentiation. This case represents the first documented case of a chondrosarcoma with differentiation into a rhabdomyosarcomatous component.


Comparison of intestinal glucose flux and electrogenic current demonstrates two absorptive pathways in pig and one in Nile tilapia and rainbow trout.
The mucosal-to-serosal flux of 14C 3-O-methyl-d-glucose was compared against the electrogenic transport of d-glucose across ex vivo intestinal segments of Nile tilapia, rainbow trout, and pig in Ussing chambers. The difference in affinities (Km "fingerprints") between pig flux and electrogenic transport of glucose, and the absence of this difference in tilapia and trout, suggest two absorptive pathways in the pig and one in the fish species examined. More specifically, the total mucosal-to-serosal flux revealed a super high-affinity, high-capacity (sHa/Hc) total glucose transport system in tilapia; a super high-affinity, low-capacity (sHa/Lc) total glucose transport system in trout and a low-affinity, low-capacity (La/Lc) total glucose transport system in pig. Comparatively, electrogenic glucose absorption revealed similar Km in both fish species, with a super high-affinity, high capacity (sHa/Hc) system in tilapia; a super high-affinity/super low-capacity (sHa/sLc) system in trout; but a different Km fingerprint in the pig, with a high-affinity, low-capacity (Ha/Lc) system. This was supported by different responses to inhibitors of sodium-dependent glucose transporters (SGLTs) and glucose transporter type 2 (GLUT2) administered on the apical side between species. More specifically, tilapia flux was inhibited by SGLT inhibitors, but not the GLUT2 inhibitor, whereas trout lacked response to inhibitors. In contrast, the pig responded to inhibition by both SGLT and GLUT2 inhibitors with a higher expression of GLUT2. Altogether, it would appear that two pathways are working together in the pig, allowing it to have continued absorption at high glucose concentrations, whereas this is not present in both tilapia and trout.


Late Pennsylvanian carbonate platform facies and coral reef: new insights from southern China (Guizhou Province).
The Pennsylvanian is characterized by intense paleoenvironmental changes related to glacio-eustatic sea-level fluctuations and major tectonic events, which affected the evolution of biocommunities. Most known Pennsylvanian tropical reefs and mounds are predominantly composed of calcareous algae (e.g. phylloid algae, Archaeolithophyllum), calcareous sponges, fenestrate bryozoans, Tubiphytes, and microbialites. However, in Houchang (southern China), the Late Pennsylvanian carbonate platform records a large coral reef lacking any analogs in age (Gzhelian), size (80-100 m thick) and composition (high biodiversity). The large coral reef developed at the border of the Luodian intraplatform basin. The intraplatform basin is characterized by the deposition of green algal grainstone, coated grain grainstone and bioclastic packstone, grainstone, floatstone and rudstone in shallow-waters. In the deep-water shelf, lithofacies are composed of burrowed bioclastic wackestone, microbioclastic peloidal packstone, grainstone, and fine-grained burrowed wackestone and packstone. In this context, the coral reef developed on a deep-shelf margin, in a moderate to low energy depositional environment, below the FWWB. The scarcity of Pennsylvanian coral reefs suggests global unfavorable conditions, which can be attributed to a complex pattern of several environmental factors, including seawater chemistry (aragonite seas), paleoclimatic cooling related to continental glaciation, and the biological competition with the more opportunistic and adaptive phylloid algal community that occupied similar platform margin paleoenvironments. The existence of the large Bianping coral reef in southern China, as well as a few additional examples of Pennsylvanian coralliferous bioconstructions, provides evidence that coral communities were able to endure the Late Paleozoic fluctuating paleoenvironmental conditions in specific settings. One of such settings appears to have been the deep shelf margin, where low light levels decreased competition with the phylloid algal community.


Synthesis of core-crosslinked zwitterionic polymer nano aggregates and pH/Redox responsiveness in drug controlled release.
A new kind of core-crosslinked zwitterionic polymer nano aggregates was prepared by two steps: (1) terpolymers of lipoic acid, polyethylene glycol diglycidyl ether and l-lysine were prepared, forming nano aggregates with hydrodynamic radius of 80 nm-183 nm by self-assembly; (2) the crosslinking of the nano aggregates took place through a reaction of side chain group in lipoic acid structural unit with 1,4-dimercaptothreotol, producing zwitterionic polymer core-crosslinked nano aggregates. The aggregates were stable against dilution and protein adsorption, and they demonstrated good pH/redox dual-responsiveness due to the introduction of disulfide bonds and zwitterionic groups to the nano aggregates. Doxorubicin (DOX) was loaded into the nano aggregates for controlled release. The drug-loaded nano aggregates exhibited obvious pH and reduction sensitivities in response to the environmental conditions in tumor cells. The nano aggregates were biocompatible and could be potentially applied as anticancer drug vehicles for enhancement of cellular uptake of anticancer drugs.


Chiral halogen and chalcogen bonding receptors for discrimination of stereo- and geometric dicarboxylate isomers in aqueous media.
Novel chiral halogen and chalcogen bonding receptors exhibit different selectivities for stereo- and geometric dicarboxylate isomers compared to a hydrogen bonding analogue. The unique geometric and electronic properties of the chalcogen bonding receptor facilitate the diagnostic fluorescence sensing of geometric dicarboxylate isomer guest species.


Potential role of the cannabinoid receptor CB1 in rapid eye movement sleep rebound.
Sleep is an unavoidable activity of the brain. The delay of the time to sleep (sleep deprivation), induces an increase of slow-wave sleep and rapid-eye-movement (REM) sleep (rebound) once the subject is allowed to sleep. This drive to sleep has been hypothesized to be dependent on the accumulation of sleep-inducing molecules and on the high expression of these molecule receptors. In this study we selectively deprived rats of REM sleep for 24 h by using the flowerpot technique. One group deprived of REM sleep was treated with SR141716A, a cannabinoid receptor 1 (CB1) receptor antagonist and then allowed to sleep for the next 4 h. Two other groups were killed, one immediately after the REM sleep deprivation period and the other after 2 h of REM sleep rebound (REM sleep deprivation plus 2 h of rebound). In both groups we determined the expression of the CB1 receptor and its mRNA. Results indicated that SR141716A prevents REM sleep rebound and REM sleep deprivation does not modify the expression of the CB1 protein or mRNA. However, REM sleep deprivation plus 2 h of sleep rebound increased the CB1 receptor protein and, slightly but significantly, decreased mRNA expression. These results suggest that endocannabinoids may be participating in the expression of REM sleep rebound.


Telmisartan decreases inflammation by modulating TNF-α, IL-10, and RANK/RANKL in a rat model of ulcerative colitis.
Telmisartan is an antihypertensive angiotensin II receptor blocker. This antihypertensive shows antiinflammatory activity. In this study, the antiinflammatory activity of telmisartan was tested in an acetic acid (10%) model of ulcerative colitis (UC) in rats. Rats were given 1, 3, and 5mg/kg/day of telmisartan orally for 3 days before induction of UC. The same doses were also administered 2 and 24h after induction. Rats from the non-colitis and non-treated colitis groups were administered vehicle (saline, 5 ml/kg) orally and another group received sulfasalazine (50mg/kg/day). Colons tissue was analyzed by macroscopic, by histopathology, by the immunohistochemical examination of RANKL/RANK pathway; by ELISA analysis of the levels of IL-10, TNF-α, myeloperoxidase (MPO) and malonaldehyde (MDA). Telmisartan at 5mg/kg reduced levels of MPO, MDA, TNF-α and increased of IL-10 (p<0.05). Additionally, telmisartan reduced macroscopic damage, number of ulcers, and inflammatory and histopathological processes such as neutrophil infiltration, changes in cytoarchitecture, and necrosis. Immunohistochemistry revealed down-regulation of nuclear factor-kappaB receptor/nuclear factor-kappaB ligand (RANK/RANKL) in groups treated with sulfasalazine or telmisartan. Telmisartan exerts beneficial effects in an acetic acid model of colitis in rats. These effects may be due to accelerated termination of the acute inflammatory phase, indicated by decreased TNF-α and increased production of IL-10 and low expression of RANKL and RANK.


The motivation and pleasure deficits but not expressivity affects social functioning through cognitive function in male patients with schizophrenia: A structural equation model.
This study aims to compare cognitive function and social functioning in male schizophrenia patients with deficit syndrome (DS) and non-DS, and to explore the associations among two different dimensions of negative symptoms (motivation and pleasure (MAP) and expressivity (EXP) deficits), cognitive function and social functioning base on a Structural Equation Model (SEM). The current study enrolled 161 male schizophrenia patients and 120 age- and education- matched healthy controls. The DS and non-DS group were categorized by the Chinese version of Schedule for the Deficit Syndrome (SDS). The psychotic and negative symptoms were evaluated by the Brief Psychiatric Rating Scale (BPRS) and the Brief Negative Symptoms Scale (BNSS). The Social functioning was measured by Scale of Social function in Psychosis Inpatients (SSPI). A battery of classical neurocognitive tests was used for assessing cognition including sustained vigilance/attention, cognitive flexibility, ideation fluency and visuospatial memory. Our study indicated that DS patients performed worser in cognitive function and social functioning than non-DS patients. The SEM model demonstrated that MAP significantly affected social functioning through direct influence and mediation of cognitive function. However, our results found that EXP had little influence on cognitive function and social function. Our findings provided evidence supporting that DS may represent as a subtype of schizophrenia, and the MAP factor play a pivotal role to influence the cognitive and social functioning in schizophrenia patients.


[The role of protein kinase C in the process of HL-60 cells induced into dendritic cells by calcium ionophore A23187].
To Investigate the role of protein kinase C (PKC) signal transduction in the process of HL-60 leukemic cells induced into dendritic cells (DCs) by calcium ionophore A23187. HL-60 cells were pretreated with protein kinase C (PKC) inhibitor Bis-1 for 24 hours followed by cultured with A23187 for another 36 hours, and the morphology, immunophenotype and function of stimulating proliferation of allogeneic T cells were compared with the cells only treated with A23187. The morphological changes, surface marker expressions and ability to stimulating the proliferation of allogeneic T cells were inhibited in DCs derived from HL-60 cells treated with PKC inhibitor Bis-1. The percentage of CD83, CD80, CD86 expressing on DCs induced by A23187 significantly decreased to (28.97 +/- 4.05)% vs. (13.23 +/- 2.15)%, (19.10 +/- 5.46)% vs. (9.70 +/- 1.69)%, (41.03 +/- 6.43)% vs. (23.37 +/- 7.50)%, respectively (P < 0.05). The induction of HL-60 cells induced into DCs by A23187 can be inhibited by PKC inhibitor Bis-1, this suggested that PKC signal transduction pathway might be involved in the process of HL-60 leukemic cells differentiated into DCs by A23187 induction.


Carotid cutdown for mechanical thrombectomy in the setting of intravenous tissue plasminogen activator: A technical report.
Acute ischemic stroke (AIS) is a leading cause of mortality and morbidity in the world. Patients are usually treated with endovascular methods and access is often gained trans-femoral or trans-radial. In patients with difficult anatomy, in rare cases the approach must be done trans-cervical. Our case illustrates a patient who received IVTPA prior to attempted endovascular treatment, leading to a direct carotid cutdown for mechanical thrombectomy. Our patient presented with a left M1 occlusion and had received intravenous thrombolytic at the presentation of his symptoms. Due to unfavorable anatomy and tortuous aortic arch, conventional access could not be obtained. In order to achieve reperfusion while maintaining hemostasis, a decision was made to perform an open carotid cutdown to catherize the left internal carotid artery to successfully retrieve the thrombus. This resulted in a TICI2B revascularization. This is the first reported case of direct carotid access in the setting of acute ischemic stroke with IVTPA. Open technique allowed us to maintain hemostasis while bypassing the tortuous arch and achieving reperfusion and give the patient the best chance for a functional recovery.


[Concept and historical evolution of epilepsy in pre-Colombian and Viceregency Peru].
We review and discuss beliefs regarding epilepsy, known in the Quechua language as sonko-nanay (disease of the heart). An overall view given by the indigenous people allows an outline to be drawn of tradition in Peru. This includes Quechua mythological beliefs, with understandable names and/or analogy with the different types of crises according to modern diagnostic classification, methods of treatment under the three different systems of government as well as real and fictitious surgery by trepanation of the skull. During the period of the Viceroys (based on Spanish medical knowledge) the different meanings are considered together with the confusion with tetanus and hysteria, and the outlines of treatment used.


Prediction of East Asian Brain Age using Machine Learning Algorithms Trained With Community-based Healthy Brain MRI.
Magnetic resonance imaging (MRI) helps with brain development analysis and disease diagnosis. Brain volumes measured from different ages using MRI provides useful information in clinical evaluation and research. Therefore, we trained machine learning models that predict the brain age gap of healthy subjects in the East Asian population using T1 brain MRI volume images. In total, 154 T1-weighted MRIs of healthy subjects (55-83 years of age) were collected from an East Asian community. The information of age, gender, and education level was collected for each participant. The MRIs of the participants were preprocessed using FreeSurfer(https://surfer.nmr.mgh.harvard.edu/) to collect the brain volume data. We trained the models using different supervised machine learning regression algorithms from the scikit-learn (https://scikit-learn.org/) library. The trained models comprised 19 features that had been reduced from 55 brain volume labels. The algorithm BayesianRidge (BR) achieved a mean absolute error (MAE) and r squared (R2) of 3 and 0.3 years, respectively, in predicting the age of the new subjects compared to other regression methods. The results of feature importance analysis showed that the right pallidum, white matter hypointensities on T1-MRI scans, and left hippocampus comprise some of the essential features in predicting brain age. The MAE and R2 accuracies of the BR model predicting brain age gap in the East Asian population showed that the model could reduce the dimensionality of neuroimaging data to provide a meaningful biomarker for individual brain aging.


Validation and clinical use of the CECA, a disease-specific quality of life questionnaire for patients with anogenital condylomata acuminata.
The aim of this validation study was to assess the measurement properties of the CECA (Spanish acronym for the Specific Questionnaire for Condylomata Acuminata) in patients with anogenital condylomas. A total of 247 patients aged > 18 years completed the questionnaire on 2 occasions as well as the Dermatology Life Quality Index (DLQI). The CECA questionnaire showed good internal consistency (Cronbach's alpha values of 0.86 and 0.91 in the emotional and sexual activity dimensions) and good testretest reliability (intraclass correlation coefficient 0.76 emotional dimension, 0.82 sexual activity dimension). Patients with de novo lesions and those with more extensive lesions and larger number of warts showed poorer health-related quality of life. CECA and DLQI scores correlated moderately. Patients whose lesions cleared at follow-up or with a reduction of >or= 50% showed a better improvement of health-related quality of life. The CECA questionnaire is a valid, reliable and sensitive tool for the assessment of health-related quality of life in patients with anogenital warts.


The use of povidone iodine in neonatal bowel surgery.
The ostomy sites of four premature infants undergoing bowel reanastomosis were lavaged with a 10% solution of povidone iodine to reduce local bacterial contamination and the risk of anastomosis dehiscence. There was a significant decrease in serum thyroxin from 112 (+/- 11) mumol/L to 90 (+/- 33) mumol/L 24 hours following surgery (P less than .05), but no change in serum thyroid stimulating hormone (TSH). The total serum iodine rose from 1.5 (+/- 0.05) mumol/L before surgery to 61.6 (+/- 46.4) mumol/L 24 hours later and urinary iodine excretion was 60 times the preoperative value in the second 24 hours following surgery. Both the hormonal and biochemical indices returned to normal 2 weeks following surgery except for total serum iodine, which remained slightly elevated. Povidone iodine is a safe and effective antiseptic agent. However, as it may cause transient suppression of thyroid function in neonates, thyroid status should be tested in all such infants at approximately 2 weeks following repeated or widespread use.


Evaluation of ability for basic movement scale for children type T (ABMS-CT) for disabled children.
The objective of this pilot study was to test the validity and reliability of a new scale, the Ability for Basic Movement Scale for Children Type T (ABMS-CT). Forty-nine pediatric patients with disabilities (aged 1.00-15.17years; 29 males and 20 females) participated in this prospective study. To prove the validity and reliability of the ABMS-CT, subjects were administered the ABMS-CT by two physicians. In addition to the ABMS-CT score, data on age, diagnosis, and results of the Functional Independence Measure for Children (WeeFIM) were recorded. Spearman's rank correlation coefficient analysis showed that the ability to perform basic movements according to the individual scores for each item on the ABMS-CT and the total scores of the ABMS-CT correlated significantly with the total scores of the motor and cognitive WeeFIM, respectively (r=0.753-0.892, p=0.0001). The five items on the ABMS-CT had appropriate internal consistency (Cronbach's α=0.966). Inter-rater reliability analysis indicated that the "oral and facial area", "hands and fingers", "one leg", "both legs", and "stairs" items on the ABMS-CT had almost perfect reliability (κ=0.854-0.925). This study provides evidence for the validity and reliability of the ABMS-CT with regard to assessment of the functional ability for complex movements in disabled pediatric patients even if they can walk independently.


Highly efficient removal of Se(IV) using reduced graphene oxide-supported nanoscale zero-valent iron (nZVI/rGO): selenium removal mechanism.
Se(IV) removal using nanoscale zero-valent iron (nZVI) has been extensively studied. Still, the synergistic removal of Se(IV) by reduced graphene oxide-supported nanoscale zero-valent iron (nZVI/rGO) has not been reported. In this study, nZVI/rGO was successfully synthesized for Se(IV) removal from wastewater. The effects of different environmental conditions (load ratio, dosage, initial pH) on Se(IV) removal by nZVI/rGO were investigated. When the load ratio is 10%, the dosage is 0.3 g/L, the initial pH is 3, and the removal rate is 99%. The adsorption isotherm and kinetics accorded with the Langmuir isotherm and first-order kinetics models (R2 > 0.99). The fitted maximum adsorption capacity reached up to 173.53 mg/g. NZVI/rGo and Se(IV) is a spontaneous endothermic reaction (△G < 0, △H > 0) and is characterized by EDS, XRD, and XPS before and after the reaction, to further clarify the reaction mechanism. The XPS narrow spectrum analysis suggested that Se(IV) was reduced to elemental selenium (Se(0)), while the intermediate Fe(II) was oxidized to form hydroxide precipitation. Therefore, nZVI/rGO was favored for Se-contaminated wastewater remediation.


Investigations on oxygen limitations of adherent cells growing on macroporous microcarriers.
Macroporous microcarriers are commonly applied to fixed and fluidized bed bioreactors for the cultivation of stringent adherent cells. Several investigations showed that these carriers are advantageous in respect to a large surface area (Griffiths, 1990; Looby, 1990a).When growing a rC-127 cell line on Cytoline 2 (Pharmacia Biotech), no satisfactory product yield could be achieved. A possible limitation in the supply of nutrient components was investigated to explain these poor results. No significant concentration gradients could be detected. Nevertheless, fluorescence staining revealed a decreasing viability, particularly inside the macroporous structure. Therefore, oxygen transfer to and into the carriers was examined by means of an oxygen microprobe during the entire process. Additional mathematical modeling supported these results.The maximum penetration depth of oxygen was determined to be 300 μm. A critical value influencing the oxygen uptake rate of the rC-127 cells occured at a dissolved oxygen concentration of 8% of air saturation. A significant mass transfer resistance within a laminar boundary film at the surface of the carrier could be detected. This boundary layer had a depth of 170 μm. The results showed that even a 40% air saturation in the bulk liquid could not provide an efficient oxygenation of the surface biofilm during the exponential growth phase. Fluorescent staining reveals a poor viability of cells growing inside the carrier volume. Thus, oxygen supply limits the growth of rC-127 cells on macroporous microcarriers. Poor process performance and low product yield could be explained this way.


Metabolic imaging of rat brain during pharmacologically-induced tinnitus.
Although much is known about the perceptual characteristics of tinnitus, its neural origins remain poorly understood. We investigated the pattern of neural activation in central auditory structures using positron emission tomography (PET) imaging in a rat model of salicylate-induced tinnitus. Awake rats were injected with the metabolic tracer, fluorine-18 fluorodeoxyglucose (FDG), once in a quiet state (baseline) and once during salicylate-induced tinnitus. Tinnitus was verified using a behavioral technique. Brain imaging was performed using a high-resolution microPET scanner. Rats underwent magnetic resonance imaging (MRI) and reconstructed MRI and microPET images were fused to identify brain structures. FDG activity in brain regions of interest were quantified and compared. MicroPET imaging showed that FDG activity in the frontal pole was stable between baseline and tinnitus conditions, suggesting it was metabolically inert during tinnitus. Inferior colliculi (p=0.03) and temporal cortices (p=0.003) showed significantly increased FDG activity during tinnitus relative to baseline; activity in the colliculi and temporal cortices increased by 17%+/-21% and 29%+/-20%, respectively. FDG activity in the thalami also increased during tinnitus, but the increase did not reach statistical significance (p=0.07). Our results show increased metabolic activity consistent with neuronal activation in inferior colliculi and auditory cortices of rats during salicylate-induced tinnitus. These results are the first to show that microPET imaging can be used to identify central auditory structures involved in tinnitus and suggest that microPET imaging might be used to evaluate the therapeutic potential of drugs to treat tinnitus.


Left hemispheric overactivation in schizophrenia: relationship to clockwise circling.
Schizophrenic patients, nonschizophrenic patients, and nonpsychiatric controls were compared in their demonstration of clockwise circling (CC), and in lateral eye movement responses to verbal and spatial questions. Only schizophrenic subjects who did not evidence CC showed inappropriate left hemisphere arousal to spatial stimuli. A tendency was noted for CC to occur more frequently among paranoid than nonparanoid schizophrenics. The results, taken together with other recent findings, suggest that the absence of CC may correlate with a more severe form of schizophrenia, and one more likely to be characterized by overarousal of the left cerebral hemisphere.


Whole-cell (+)-ambrein production in the yeast Pichia pastoris.
The triterpenoid (+)-ambrein is a natural precursor for (-)-ambrox, which constitutes one of the most sought-after fragrances and fixatives for the perfume industry. (+)-Ambrein is a major component of ambergris, an intestinal excretion of sperm whales that is found only serendipitously. Thus, the demand for (-)-ambrox is currently mainly met by chemical synthesis. A recent study described for the first time the applicability of an enzyme cascade consisting of two terpene cyclases, namely squalene-hopene cyclase from Alicyclobacillus acidocaldarius (AaSHC D377C) and tetraprenyl-β-curcumene cyclase from Bacillus megaterium (BmeTC) for in vitro (+)-ambrein production starting from squalene. Yeasts, such as Pichia pastoris, are natural producers of squalene and have already been shown in the past to be excellent hosts for the biosynthesis of hydrophobic compounds such as terpenoids. By targeting a central enzyme in the sterol biosynthesis pathway, squalene epoxidase Erg1, intracellular squalene levels in P. pastoris could be strongly enhanced. Heterologous expression of AaSHC D377C and BmeTC and, particularly, development of suitable methods to analyze all products of the engineered strain provided conclusive evidence of whole-cell (+)-ambrein production. Engineering of BmeTC led to a remarkable one-enzyme system that was by far superior to the cascade, thereby increasing (+)-ambrein levels approximately 7-fold in shake flask cultivation. Finally, upscaling to 5 L bioreactor yielded more than 100 mg L-1 of (+)-ambrein, demonstrating that metabolically engineered yeast P. pastoris represents a valuable, whole-cell system for high-level production of (+)-ambrein.


Protective or Inhibitory Effect of Pharmacological Therapy on Cardiac Ischemic Preconditioning: A Literature Review.
Ischemic preconditioning (IP) is an innate phenomenon, triggered by brief, non-lethal cycles of ischemia/reperfusion applied to a tissue or organ that confers tolerance to a subsequent more prolonged ischemic event. Once started, it can reduce the severity of myocardial ischemia associated with some clinical situations, such as percutaneous coronary intervention (PCI) and intermittent aortic clamping during coronary artery bypass graft surgery (CABG). Although the mechanisms underlying IP have not been completely elucidated, several studies have shown that this phenomenon involves the participation of cell triggers, intracellular signaling pathways, and end-effectors. Understanding this mechanism enables the development of preconditioning mimetic agents. It is known that a range of medications that activate the signaling cascades at different cellular levels can interfere with both the stimulation and the blockade of IP. Investigations of signaling pathways underlying ischemic conditioning have identified a number of therapeutic targets for pharmacological manipulation. This review aims to present and discuss the effects of several medications on myocardial IP.


Mycotic aortic aneurysm caused by haemophilus influenzae group F.
Haemophilus influenzae is a rare cause of mycotic aortic aneurysm. We present a case of H. influenzae mycotic aortic aneurysm, which was complicated by prior endovascular stent-graft placement at another facility. A 58 year-old man was treated by endograft placement for a presumed penetrating aortic ulcer after having symptoms of abdominal pain and malaise for one month. He presented to our institution 11 days after endograft placement with septic physiology. Repeat computed tomography angiogram demonstrated an inflammatory mass around the distal aorta and right common iliac artery, which had an associated contained rupture. The patient was treated using intravenous antibiotics, axillo-bifemoral bypass followed by endograft explantation and aortic and iliac ligation. Intraoperative cultures grew Haemophilus influenzae, serotype f. Aortic endografts have been successfully used for treatment of selected mycotic aneurysms, generally after adequate treatment of the primary infection with intravenous antibiotics. This case demonstrates the unfavorable natural history of endograft placement in an unsuspected mycotic aneurysm. A high index of suspicion for possible aortic infection should be maintained for patients with systemic symptoms and unusual aortic pathology prior to choosing endovascular repair.


The influence of light path length on the color of synthetic ruby.
The corrected ultraviolet-visible light spectrum was used to calculate the color of synthetic rubies with different light path lengths, and the influence of light path length and standard light source on the color of synthetic ruby was studied. The results show that the difference in colour between the o light direction and the e light direction of the synthetic ruby decreases as the length of the light path increases. At the same time, as the length of the light path increases, the lightness L* decreases, and the hue angle h° increases. The chroma C* first increases as the length of the light path increases, and then begins to decrease under the influence of the continuous decrease in lightness. The color difference ΔE*ab reaches the maximum when the light path length is around 10 mm, and the standard light source has the greatest influence on the color difference ΔE*ab. As the length of the light path continues to increase, the influence of the standard light source on the color difference ΔE*ab decreases. In the ultraviolet-visible light spectrum, the strong absorption band of Cr3+ at 545 nm is the main cause of the color of the ruby. The larger the area of the band at 545 nm, the lower the lightness and the higher the hue angle, which means the ruby colour is redder.


K(ATP) channel pharmacogenomics: from bench to bedside.
Inheritance plays a significant role in defining drug response and toxicity. Advances in molecular pharmacology and modern genomics emphasize genetic variation in dictating inter-individual pharmacokinetics and pharmacodynamics. A case in point is the homeostatic ATP-sensitive potassium (K(ATP)) channel, an established drug target that adjusts membrane excitability to match cellular energetic demand. There is an increased recognition that genetic variability of the K(ATP) channel impacts therapeutic decision-making in human disease.


Whole body gestational donation.
Whole body gestational donation offers an alternative means of gestation for prospective parents who wish to have children but cannot, or prefer not to, gestate. It seems plausible that some people would be prepared to consider donating their whole bodies for gestational purposes just as some people donate parts of their bodies for organ donation. We already know that pregnancies can be successfully carried to term in brain-dead women. There is no obvious medical reason why initiating such pregnancies would not be possible. In this paper, I explore the ethics of whole-body gestational donation. I consider a number of potential counter-arguments, including the fact that such donations are not life-saving and that they may reify the female reproductive body. I suggest if we are happy to accept organ donation in general, the issues raised by whole-body gestational donation are differences of degree rather than substantive new concerns. In addition, I identify some intriguing possibilities, including the use of male bodies-perhaps thereby circumventing some potential feminist objections.


[Functional and morphologic changes in the heart following phrenicotomy].
Experiments on dogs showed that the phrenic nerves participate in innervation of the heart. Their division in the cervical portion is attended with disorders of the rhythm, abnormalities in the components of the electrocardiogram, and degenerative changes in the intramural nerve apparatus of the heart. The results of the experiments indicate that the use of the phrenic nerve as a donor in a plastic operation of the nerve trunks should be considered an operation of choice.


Genetic manipulation of secondary metabolite biosynthesis for improved production in Streptomyces and other actinomycetes.
Actinomycetes continue to be important sources for the discovery of secondary metabolites for applications in human medicine, animal health, and crop protection. With the maturation of actinomycete genome mining as a robust approach to identify new and novel cryptic secondary metabolite gene clusters, it is critical to continue developing methods to activate and enhance secondary metabolite biosynthesis for discovery, development, and large-scale manufacturing. This review covers recent reports on promising new approaches and further validations or technical improvements of existing approaches to strain improvement applicable to a wide range of Streptomyces species and other actinomycetes.


Isolation, cDNA sequences, and biochemical characterization of the major cyclosporin-binding proteins of Toxoplasma gondii.
The activities of the immunosuppressive, antifungal compounds cyclosporin A (CsA), FK-506, and rapamycin are dependent upon high affinity binding proteins collectively termed immunophilins. We report the isolation, biochemical characterization, and amino acid sequences of two major CsA-binding proteins, cyclophilins, from the pathogenic protozoan, Toxoplasma gondii. The 18.5- and 20-kDa molecular mass proteins exhibit peptidylproline cis-trans-isomerase activity, which is inhibitable by 10(-8) M CsA. The amino acid sequences of these two proteins, deduced from cDNA clones, reveal up to 70% amino acid identity to previously isolated cyclophilins. The 18.5-kDa protein appears to be synthesized as a precursor with a 15 amino acid signal peptide. The amino-terminal region of the mature 20-kDa protein has significant homology to the B subunit of the calmodulin-dependent phosphatase, calcineurin. The two T. gondii cyclophilins are products of different genes and appear to have different subcellular distributions.


Genetic contribution to iron status: SNPs related to iron deficiency anaemia and fine mapping of CACNA2D3 calcium channel subunit.
Numerous studies associate genetic markers with iron- and erythrocyte-related parameters, but few relate them to iron-clinical phenotypes. Novel SNP rs1375515, located in a subunit of the calcium channel gene CACNA2D3, is associated with a higher risk of anaemia. The aim of this study is to further investigate the association of this SNP with iron-related parameters and iron-clinical phenotypes, and to explore the potential role of calcium channel subunit region in iron regulation. Furthermore, we aim to replicate the association of other SNPs reported previously in our population. We tested 45 SNPs selected via systematic review and fine mapping of CACNA2D3 region, with haematological and biochemical traits in 358 women of reproductive age. Multivariate analyses include back-step logistic regression and decision trees. The results replicate the association of SNPs with iron-related traits, and also confirm the protective effect of both A allele of rs1800562 (HFE) and G allele of rs4895441 (HBS1L-MYB). The risk of developing anaemia is increased in reproductive age women carriers of A allele of rs1868505 (CACNA2D3) and/or T allele of rs13194491 (HIST1H2BJ). Association of SNPs from fine mapping with ferritin and serum iron suggests that calcium channels could be a potential pathway for iron uptake in physiological conditions.


An isolated left common carotid artery from the main pulmonary artery in a neonate with aortic valve atresia.
A 20-day-old girl was referred to our clinic for systolic murmur, cyanosis, and dyspnoea with feeding. Echocardiography revealed an atretic aortic valve. CT angiography scan revealed that the left common carotid artery originated from the distal main pulmonary artery. The plan was patent ductus artriosus stenting and bilateral pulmonary artery banding and then follow-up for any possible future intervention.


Light- and electron-microscopic studies on the gastric parietal cells in perinatal rats. Effects of fetal and neonatal oral administration of milk.
Perinatal changes of the gastric parietal cells were studied under normal and various experimental conditions. Histologic examination revealed that the parietal cells appeared markedly increased in number from late fetal to early neonatal days. When premature newborn rats were delivered by cesarean section and were nursed by foster mothers for 1 or 2 days, the degree of increase of these cells reached nearly that in the normal neonates. On the other hand, in fetuses retained in utero 1 day beyond the normal gestation, these cells appeared not increased in number. When these postmature fetuses were delivered, the parietal cells turned to increase in number in a degree similar to that in the normal neonates. Milk given to the premature newborn rats caused 6 h later a marked increase in the number of the parietal cells. Milk given to fetuses in utero also caused an increase of these cells. The results suggest that the development of the parietal cells in perinatal rats is accelerated by the intake of milk and that the fetal parietal cells are ready to respond to milk given at least 1 day before birth.


Ambulatory blood pressure, proteinuria and uric acid in children with IgA nephropathy and their correlation with histopathological findings.
In adults, nighttime hypertension and hyperuricemia are new risk factors for progression of IgA nephropathy (IgAN). In children, nighttime blood pressure (BP) and serum uric acid have never been investigated. The aim of the study was to investigate nighttime BP and uric acid in children with IgAN. Data on children with IgAN from two pediatric nephrology centers were retrospectively reviewed (renal biopsy - subclasses according Hass I-V, ambulatory blood pressure monitoring ABPM, serum uric acid, proteinuria). Twenty-eight untreated children with IgAN were included. Hypertension was diagnosed on the basis of ABPM in 54% of children, 50% were nondippers and 25% have isolated nighttime hypertension. The mean ambulatory BP was higher at nighttime than during daytime (systolic nighttime BP 1.11 +/- 0.79 SDS vs. daytime 0.59 +/- 0.79, diastolic nighttime BP 1.16 +/- 0.95 vs. daytime 0.52 +/- 1.10, p < 0.01 for systolic and p = 0.01 for diastolic). Children with severe histological subclasses (III-IV) tended to have higher prevalence of hypertension than children with mild subclasses (I-II), 67% vs. 38%, p = 0.13. Hyperuricemia was diagnosed in 14% of children. A significant correlation was found between proteinuria and histopathological subclasses (r = 0.44, p < 0.05). Children with IgAN have often nighttime hypertension. Hypertension and proteinuria are associated with severe histopathological findings. Hyperuricemia is a rare finding in children.


The U Shape of Prostate-specific Antigen and Prostate Cancer-specific Mortality in High-grade Metastatic Prostate Adenocarcinoma.
Accumulated evidence suggests that metastatic prostate cancer (mPCa) with a low prostate-specific antigen (PSA) level may be a unique entity. However, its clinical features and prognosis have not been fully evaluated. To investigate the clinical features of low-PSA mPCa and the impact of low PSA level on overall survival (OS) and PCa-specific mortality (PCSM) of mPCa. A total of 8479 mPCa patients were retrieved from the Surveillance, Epidemiology, and End Results program (2010-2015). The median follow-up was 18 mo. Cox regression and Fine-Gray competing risk were used to calculate the hazard ratio (HR) and subdistribution hazard ratio (sHR) for OS and PCSM, respectively. A higher rate of T4 stage disease (19.8%) and visceral metastasis (18.2%) and the shortest median OS (34 mo) were observed in mPCa patients with Gleason 8-10 and PSA ≤4ng/ml. In the Cox regression model, PSA ≤4ng/ml was a significant predictor of OS for Gleason 8-10 disease. The distribution of PCSM by PSA was U-shaped for Gleason score 8-10 (PSA 4.1-10ng/ml as the referent), with an adjusted sHR of 1.52 for PSA ≤4.0ng/ml (95% confidence interval: 1.17-1.96) versus 0.99 for PSA 10.1-20ng/ml and 1.35 for PSA >20ng/ml. In contrast, the distribution of PCSM by PSA was linear for Gleason 5-7. Sensitivity analyses showed similar results in Gleason 9-10 and Gleason 10 subgroup. The study is limited by its retrospective design. Low PSA, high-grade mPCa has a higher proportion of T4 stage disease, visceral metastasis, and PCSM. We found that 2.8% of high-grade metastatic prostate cancer has a prostate-specific antigen level ≤4ng/ml at diagnosis. This population has aggressive clinical features and a poor cancer-specific outcome. Our results highlighted this under-reported population, and the management of these patients warrants further research.


A Systematic Review and Meta-Analysis Protocol on How Best to Use Non-Pharmacologic Therapies to Manage Chronic Low Back Pain and Associated Depression.
To evaluate the efficacy of different non-pharmacologic therapies (NPTs) on relieving depressive symptoms and pain intensity in individuals living with chronic low back pain (LBP) and associated depression. A comprehensive search of seven English databases and two Chinese databases from inception to the search date will be undertaken. The reference lists of previously published relevant reviews and included trials will also be searched. Only peer-reviewed and published moderate-to-high quality randomized controlled trials (RCTs) for chronic LBP and associated depression treated with NPTs will be considered. Two independent reviewers will identify studies, extract data, assess risk of bias, and evaluate the strength of evidence using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. Meta-analysis will be conducted to estimate the treatment effect of various NPTs. Heterogeneity will be assessed using Cochrane's Q and the I-squared statistics. Subgroup and sensitivity analyses will be performed to assess the robustness of findings. A funnel plot will be developed to evaluate reporting bias, and Begg's and Egger's tests will be used to assess funnel plot symmetries. This protocol outlines the planned scope and methodology for an upcoming systematic review and meta-analysis, which will provide up-to-date evidence on 1) which NPTs are associated with improvements in depressive symptoms and pain intensity and 2) whether the effects of NPTs on chronic LBP and associated depression vary according to clinical condition, participant, and treatment characteristics. Our meta-analyses of moderate-to-high quality RCTs will help to develop specific recommendations on prescribing NPTs in patients with chronic LBP and associated depression. This protocol is registered on the International Platform of Registered Systematic Review and Meta-analysis (INPLASY) protocols platform as record No. INPLASY202260055.


Genetics of Parkinson's disease.
Twenty years ago Parkinson's disease (PD) was thought of as an environmentally determined neurodegenerative disease. It is now known that there are two autosomal dominant disease genes, alpha-synuclein and dardarin, and three genes responsible for autosomal recessive PD, parkin, DJ-1 and PINK-1. Although these gene mutations are not common, their identification has led to a new understanding of the pathogenesis of PD, and to a development in the understanding of the clinical and pathological definitions of PD and Lewy body disease. Ultimately, these advances may lead to the development of new disease-modifying therapies, but more immediately these discoveries have led to a more coherent view of the spectrum of PD and Lewy body diseases and to accurate genetic diagnosis and counselling for some families.


Predictors and moderators of outcome for severe and enduring anorexia nervosa.
Few of the limited randomized controlled trials (RCTs) for adults with anorexia nervosa (AN) have explored predictors and moderators of outcome. This study aimed to identify predictors and moderators of outcome at end of treatment (EOT) and 6- and 12-month follow-up for adults with AN (N = 63). All participants met criteria for severe and enduring AN (duration of illness ≥ 7 years) and participated in an RCT of cognitive-behavioral therapy (CBT-AN) and specialist supportive clinical management (SSCM). General linear models were utilized and included all available outcome data at all time points. Outcome was assessed across three domains: eating disorder quality of life (EDQOL), mental health (MCS), and depressive symptoms (BDI). Predictors of better outcome included: lower age, shorter duration of illness, having AN-R, being employed, not taking psychotropic medication, and better social adjustment. Four moderators of treatment outcome emerged: eating disorder psychopathology (EDE Global), depression (BDI), age, and AN subtype. Participants with higher baseline scores on these measures, older age, or binge eating/purging subtype benefited more from CBT-AN than SSCM. Older patients with more severe eating-related psychopathology and depression have better outcomes in a behaviorally targeted treatment such as CBT-AN rather than a supportive treatment such as SSCM.


Overproduction and purification of Mycobacterium tuberculosis WhiB3 in Escherichia coli is enhanced by co-expression with trigger factor chaperone.
Members of the WhiB-like (Wbl) family of proteins are found in Acintomycetes and are somewhat recalcitrant to overproduction as soluble proteins in the laboratory protein expression workhorse Esherichia coli. The aim of this study was to evaluate the effects of culture conditions and co-expression of the chaperone protein, trigger factor (TF), on the soluble production of recombinant Mycobacterium tuberculosis (Mtb) WhiB3. A pET28a derived expression plasmid coding for His6-WhiB3 was created and the effects of varying the concentration of inducer (IPTG), the timing of induction, the nature of the inducer (auto-induction medium) and the temperature of the cultivation on the production of soluble His6-WhiB3 were tested. Whilst His6-WhiB3 protein was readily detected, the overwhelming majority of the protein was present in the insoluble fraction of cell-free extracts. However, co-expression of the tig from pTf16, coding for TF, increased His6-WhiB3 solubility dramatically, facilitating its isolation by affinity chromatography. Purified His6-WhiB3 was shown to be monomeric, and UV-visible spectra suggested that ∼10% of the isolated protein possessed a [4Fe-4S] cluster. The secondary structural properties of His6-WhiB3 were altered by acquisition of an iron-sulfur cluster. By developing a protocol to readily overproduce and purify WhiB3, this study paves the way for future structure-function experiments.


Brillouin propagation modes in optical lattices: interpretation in terms of nonconventional stochastic resonance.
We report the first direct observation of Brillouin-like propagation modes in a dissipative periodic optical lattice. This has been done by observing a resonant behavior of the spatial diffusion coefficient in the direction corresponding to the propagation mode with the phase velocity of the moving intensity modulation used to excite these propagation modes. Furthermore, we show theoretically that the amplitude of the Brillouin mode is a nonmonotonic function of the strength of the noise corresponding to the optical pumping, and discuss this behavior in terms of nonconventional stochastic resonance.


Three-dimensional (3D) palladium-zinc oxide nanowire nanofiber as photo-catalyst for water treatment.
Zinc Oxide Nano Wires (ZNWs) has been considered as a promising material for purification and disinfection of water and remediation of hazardous waste owing to its high activity and lower cost. In this study, three-dimensional (3D) structured palladium (Pd)/ZNWs were synthesized on the fabricated electrospun nanofibers and explored for enhancement of organic matter (OM) removal efficiency in water by suppressing electron-hole recombination during photocatalytic activity and increased surface area. The densely populated ZNWs were fabricated on the electrospun nanofiber by electroless plating (EP) and hydrothermal synthesis. In order to improve photocatalytic efficiency, a thin layer of Pd was coated prior to ZNWs growth to induce suppression of electron hole recombination produced during catalyst activity. The creation of a highly porous network of nanofibers decorated with ZNWs resulted in an increase of specific removal rate (SRR) of OM from 0.0249 to 0.0377 mg CODCr removed/mg ZNWs-hr when ZNW were grown on a Pd layer. It is believed that the demonstration of OM removal in the water through Pd/ZNWs membrane and enhanced photocatalytic activity under UV irradiation from layered structure can broaden potential applicability of Pd/ZNWs membranes for various photo catalytic water treatment.


What prevails in climatic response of Pinus sylvestris in-between its range limits in mountains: slope aspect or elevation?
The roles of slope orientation and elevational temperature gradient were investigated for Scots pine (Pinus sylvestris L.) growth in the middle of its growth range, where these factors can significantly modulate microclimate and thus plant growth. We assumed that slope orientation causes more complex and severe effects than elevation because it influences all three main factors of plant growth: light, heat, and moisture. In addition to the total ring width, the earlywood and latewood width and latewood ratio were considered variables that contain information about tree ring growth during the season and wood structure over all tree life span on three sampling sites at different elevations and opposite slopes. Despite the observed dependence of pine growth rate on temperature and solar radiation, the mean latewood ratio is stable and similar between all sampling sites, being presumably defined by the genotype of individual trees. The seasonality of the climatic response of tree growth is bound to spatiotemporal variation of the vegetative season timing due to the elevational temperature lapse and local warming. However, its direction is primarily defined by slope orientation, where southern slope is moisture-limited, even at adjacent sites, and divergent climatic reactions of earlywood (weak moisture-limited in the last decades) and latewood growth (temperature-limited) were revealed on the northern slope.


Hepatitis E virus infection and waste pickers: A case-control seroprevalence study.
Whether waste pickers are a risk group for hepatitis E virus (HEV) infection is largely unknown. This study aimed to determine the association between HEV exposure and the occupation of waste pickers and the work characteristics of waste pickers. An age-and gender-matched case-control seroprevalence study of 86 waste pickers and 86 control subjects of the general population was performed. We determined anti-HEV IgG antibodies in sera of cases and controls using a commercially available enzyme-linked immunoassay. The McNemar's test was used to assess the association between HEV seropositivity and the occupation of waste picker. The association between HEV seropositivity and work characteristics of waste pickers was assessed by bivariate and logistic regression analyses. Anti-HEV IgG antibodies were detected in 14 (16.3%) of the 86 waste pickers and in 8 (9.3%) of the 86 control subjects (McNemar's pair test: odds ratio (OR) = 13.0; 95% confidence interval (CI): 0.73-230.77; p = .02). Bivariate analysis showed that HEV exposure was associated with an ill status (p = .01) and reflexes impairment (p = .009). Logistic regression analysis showed that HEV seropositivity was associated with increasing age (OR = 6.52; 95% CI: 1.95-21.78; p = .002) and raising pigs (OR = 12.01; 95% CI: 1.48-97.26; p = .02). This is the first age- and gender-matched case-control study on the association between HEV infection and the occupation of waste picker. Waste pickers represent a risk group for HEV infection. Factors associated with HEV seropositivity found in this study may help in the design of optimal planning to avoid HEV infection.


Impact of fundus-first laparoscopic cholecystectomy for severe cholecystitis.
The Tokyo Guidelines 2018 proposed fundus-first laparoscopic cholecystectomy (FFLC) as a bailout surgery. This study investigated the clinical impact of FFLC for severe cholecystitis. This study reviewed 772 patients who underwent laparoscopic cholecystectomy (LC) between 2015 and 2018. Of these patients, 171 patients were diagnosed with severe cholecystitis according to our difficulty scoring system. FFLC was not prevalent in our faculty for the first 2 years [early period group (EG)], whereas FFLC was predominantly used for the last 2 years [late period group (LG)]. There were 81 patients (47%) belonging to the EG and 90 patients (53%) in the LG. The clinical data and the surgical outcomes of these patients were retrospectively analyzed. The difficulty score did not differ between the two groups (11 vs. 11 points, p = 0.846). Patients underwent FFLC significantly more frequently in the LG (63% vs. 12%, p = 0.020). Laparoscopic subtotal cholecystectomy (LSC) was done in 10 patients (11%) of the LG, which was significantly low compared to that in the EG (n = 20, 25%) (p = 0.020). In all patients, LC was safely achieved without bile duct injury or conversion to laparotomy. The incidence of choledocholithiasis was significantly low in the LG (0 vs. 4, p = 0.048). The median postoperative hospital stay was significantly shorter in the LG (6 vs. 4 days, p < 0.001). After the introduction of FFLC, there were significant improvements in the surgical outcomes of LC for severe cholecystitis, including the rate of LSC, incidence of choledocholithiasis, and duration of postoperative hospital stay.


Role of Progenitor Cells in Liver Regeneration after Subtotal Resection.
In the liver of rats subjected to subtotal liver resection (80% organ weight), the expression of sox9 gene and SOX9 protein content increased and cells with hepatocyte morphology expressing SOX9 appeared; the proportion of cells expressing cytokeratin-19 also increased. Based on these data, we cannot completely exclude the involvement of resident progenitor cells and hepatocyte reprogramming in liver regeneration after subtotal resection, however, the contribution of these processes seems to be insignificant. The leading mechanism of liver mass recovery after subtotal resection is proliferation of hepatocytes.


Phylogenetic evidence of the transfer of nodZ and nolL genes from Bradyrhizobium to other rhizobia.
Nod factor modifications mediated by nodZ and nolL gene products (fucosylation and acetylation of fucose residues, respectively) were probably later acquisitions in the nodulation process. Novel phylogenetic analyses suggest that nodZ and nolL genes were transferred from Bradyrhizobium to other nodule bacteria. These bradyrhizobial genes are highly diverse while rhizobial, sinorhizobial and mesorhizobial nodZ and nolL genes are represented by few branches among those from bradyrhizobia. These genes in novel rhizobial backgrounds may have favored efficient nodulation in legume hosts commonly associated with Bradyrhizobium strains.


Neural and functional correlates of impaired reading ability in schizophrenia.
Deficits in early auditory processing (EAP) are a core component of schizophrenia (SZ) and contribute significantly to impaired overall function. Here, we evaluate the potential contributions of EAP-related impairments in reading to functional capacity and outcome, relative to effects of auditory social cognitive and general neurocognitive dysfunction. Participants included 30-SZ and 28-controls of similar age, sex, and educational achievement. EAP was assessed using an auditory working memory (tone-matching) task. Phonological processing and reading Fluency were assessed using the Comprehensive Test of Phonological Processing and Woodcock-Johnson reading batteries, respectively. Auditory-related social cognition was assessed using measures of emotion/sarcasm recognition. Functional capacity and outcome were assessed using the UCSD Performance-based Skills Assessment and Specific Level of Functioning scale, respectively. fMRI resting-state functional-connectivity (rsFC) was used to evaluate potential underlying substrates. As predicted, SZ patients showed significant and interrelated deficits in both phonological processing (d = 0.74, p = 0.009) and reading fluency (d = 1.24, p < 0.00005). By contrast, single word reading (d = 0.35, p = 0.31) was intact. In SZ, deficits in EAP and phonological reading ability significantly predicted reduced functional capacity, but not functional outcome. By contrast, deficits in reading fluency significantly predicted impairments in both functional capacity and functional outcome. Moreover, deficits in reading fluency correlated with rsFC alterations among auditory thalamus, early auditory and auditory association regions. These findings indicate significant contributions of EAP deficits and functional connectivity changes in subcortical and early auditory regions to reductions in reading fluency, and of impaired reading ability to impaired functional outcome in SZ.


Virome characterization of Cryphonectria parasitica isolates from Azerbaijan unveiled a new mymonavirus and a putative new RNA virus unrelated to described viral sequences.
Cryphonectria parasitica, the causal agent of chestnut blight is controlled in Europe through natural spread of Cryphonectria hypovirus 1 (CHV1), a mycovirus able to induce hypovirulence to the host. In recent years C. parasitica was reported infecting Azerbaijani population of chestnut, but the presence of CHV1 still needs to be confirmed. Aim of this work was to investigate fifty-five C. parasitica isolates collected in Azerbaijan to describe the associated viruses. Our work found i) the first negative-sense ssRNA virus known to infect C. parasitica naturally for which we propose the name Cryphonectria parasitica sclerotimonavirus 1 (CpSV1) and ii) an RNA sequence showing peculiar features suggesting a viral nature for which we propose the name Cryphonectria parasitica ambivirus 1 (CpaV1). The discovery of CpaV1 expands our knowledge of the RNA virosphere suggesting the existence of a new lineage that cannot presently be reliably associated to the monophyletic Riboviria.


The cytotoxic effect of two chemotypes of essential oils from the fruits of Angelica archangelica L.
The aim of this work was to study the constituents and cytotoxicity of the essential oils from the fruits of Angelica archangelica growing in Iceland. Three samples of essential oils were prepared by steam distillation. Their composition was established with GC/MS. The effects of the oils were examined in PANC-1 human pancreas cancer cells and Crl mouse breast cancer cells in concentrations ranging from 10-400 microg/ml, measuring the reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl)-2- (4-sulphophenyl) -2H-tetrazolium (MTS) by mitochondrial enzymes. Two types of essential oils were found, differing mainly in the absence or presence of beta-phellandrene. The ED50 of the oils ranged from 48.6 microg/ml to 108.3 microg/ml for PANC-1 and 48.0 microg/ml to 91.8 microg/ml for Crl cells. The cytotoxic activity of the essential oils was independent of the quantity of their main components.


Synthesis and pharmacological characterization of potent, selective, and orally bioavailable isoindoline class dipeptidyl peptidase IV inhibitors.
Focused structure-activity relationships of isoindoline class DPP-IV inhibitors have led to the discovery of 4b as a highly selective, potent inhibitor of DPP-IV. In vivo studies in Wistar/ST rats showed that 4b was converted into the strongly active metabolite 4l in high yield, resulting in good in vivo efficacy for antihyperglycemic activity.


The mitochondrial DNA mutation (deltamtDNA5286) in human oocytes: correlation with age and IVF outcome.
To evaluate the correlation between the presence of a newly described mitochondrial DNA (mtDNA) mutation (deltamtDNA5286), patients' age, and in vitro fertilization (IVF) outcomes. The presence of deltamtDNA5286 was analyzed by nested-primer PCR in 224 unfertilized oocytes of 81 women undergoing IVF for various reasons. Age, number of oocytes retrieved, fertilization and embryo cleavage, number of embryos transferred, and pregnancy rates were compared between patients with and without a mtDNA mutation in their oocytes. The 23 patients in which deltamtDNA5286 was detected in at least one oocyte were significantly younger than the other 58 with no mutations (30.9 years vs. 33.8 years, respectively, P = 0.03), and had a significantly lower fertilization rate (28.9% vs. 42.1%, respectively, P = 0.01). No other outcome variable was significantly different between the two groups. The mtDNA mutation deltamtDNA5286 may serve as a marker of decreased oocyte quality in IVF.


Increased expression of Mcl-1 is responsible for the blockage of TRAIL-induced apoptosis mediated by EGF/ErbB1 signaling pathway.
Epidermal growth factor (EGF) protects against death receptor induced apoptosis in epithelial cells. Herein, we demonstrate that EGF protection against tumor necrosis factor related apoptosis-inducing ligand (TRAIL) induced apoptosis is mediated by increased expression of the Bcl-2 family member myeloid cell leukemia 1 (Mcl-1). EGF increased the mRNA and protein levels of Mcl-1. Furthermore, expression of ErbB1 alone or in combination with ErbB2 in NIH3T3 cells up-regulates Mcl-1 following EGF treatment. In addition, up-regulation of Mcl-1 by EGF is mediated through AKT and NFkappaB activation since kinase inactive AKT and DeltaIkappaB effectively blocks this up-regulation. NFkappaB was also critical for the ability of EGF to prevent TRAIL induced apoptosis as a dominant negative IkappaB (DeltaIkappaB) blocked NFkappaB activation, and relieved EGF protection against TRAIL mediated mitochondrial cytochrome-c release and apoptosis. Finally, anti-sense oligonucleotides directed against Mcl-1 effectively reduced the protein levels of Mcl-1 and blocked EGF protection against TRAIL induced mitochondrial cytochrome-c release and apoptosis. Taken together, EGF signaling leads to increased Mcl-1 expression that is required for blockage of TRAIL induced apoptosis.


Computerized tomography and surgical treatment in intracranial suppuration. Report of 30 consecutive unselected cases of brain abscess and subdural empyema.
The authors report their experience with 30 cases of intracranial suppuration: 23 with brain abscess and seven with subdural empyema. All of the cases were diagnosed by means of computerized tomography and enhancement with intravenous contrast material. Most of the patients were treated by single or repeated aspiration through burr holes.


Detection of anti-RNA antibodies in systemic lupus erythematosus by ELISA using nylon as solid support.
A highly sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) for the detection of anti-RNA antibodies is described. The assay procedure involves adsorption of total cellular RNA on nylon beads which could be conveniently stored for a considerable period of time without loss in antigenicity. Sixty-four percent of systemic lupus erythematosus (SLE) sera were positive for anti-RNA antibodies with fluorogenic substrate against 48% with colorigenic substrate.


Stage-specific production of S-antigens of Plasmodium falciparum in vitro.
Stage-specific synthesis and release of the S-antigen of Plasmodium falciparum was demonstrated using 3H-glycine-labelled, synchronous, cloned parasite populations. The release of S-antigens into culture supernatants was first evident 25 to 30 hours after initiation of the parasite cycle and correlated with the rupture of the most mature schizonts. S-antigen release reached a peak at 37 to 43 hours at the height of the reinvasion cycle. Intracellular production of S-antigens was evident slightly earlier (19 to 24 hours) and correlated with the initial appearance of schizonts.


Mutagenesis of subunit delta from Escherichia coli F1F0-ATP synthase.
In Escherichia coli, the F1 sector of the F1F0-ATP synthase is connected to the membrane-embedded F0 sector by a narrow stalk, thought to be formed by subunits delta and b. Mutagenic analysis was used here to study the structure and function of subunit delta. First, random mutations in the protein were generated by bisulfite mutagenesis. Two single missense mutations causing impaired growth by oxidative phosphorylation were found, namely delta A149T and delta G150D. Both occur at the conserved C-terminal region, which has been suggested previously to be functionally important. Two techniques were applied to study the C-terminal region in greater detail. Cassette mutagenesis was used to randomly mutate the sequence from delta 145 to delta 167, and residues delta A149 and delta G150 were specifically mutated by site-directed mutagenesis to obtain multiple substitutions at each position. Fifteen of the residues between delta 145 and delta 167 were mutated. None was found to be absolutely essential for function. However, the properties of the mutants obtained, which included partial impairment of growth by oxidative phosphorylation, temperature sensitivity, and specific structural requirements at residues delta A149 and delta G150, confirmed that this region is important for enzyme function. Based on these studies, and on secondary and tertiary structure predictions, a model for subunit delta and its orientation in F1F0-ATP synthase is proposed.


Assessment of Oxidative Stress in Patients with Sudden Hearing Loss: A Non-randomized Prospective Clinical Study.
The etiology of sudden hearing loss (SHL) has not been fully elucidated. Previous studies claimed that different etiological factors may play a role in the pathogenesis of SHL. The aim of the present study is to investigate the presence of oxidative stress (OS) in SHL cases using thiol-disulfide balance. In addition, total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI) and lipid hydroperoxide levels (LOOH) were investigated. A total of 30 cases (15 female, 15 male, mean age 48.9 ± 8.1 years, age range: 36-68 years) were included in the study. Thiol and disulfide amounts, thiol/disulfide ratios, TOS, TAS, OSI and LOOH scores of the case group and control group were compared. Native thiol (SH) and total thiol (SH + SH) values were significantly lower in the SHL group than in the control group (p = 0.028 and p = 0.044, respectively). The LOOH value, TOS value and OSI value were significantly higher in the SHL group than in the control group (all p values < 0.05). The TAS value was significantly lower in the SHL group than in the control group (p = 0.0001). The present study has presented that the thiol-disulfide balance was impaired in SHL cases. OS may play a role in the development of SHL.


Expression of five forms of microsomal cytochrome P-450 in primary cultures of rabbit hepatocytes treated with various classes of inducers.
In order to investigate the expression of five different forms of microsomal cytochrome P-450 including P-450 2 (P450IIBI), 3b (P450IIC3), 3c (P450IIIA4), 4 and 6 (P450IA2 and A1), hepatocytes were isolated from untreated rabbit and maintained in primary monolayer cultures in serum free modified Waymouth medium in the absence and in the presence of various classes of inducers including phenobarbital (PB), rifampicin (RIF), dexamethasone (DEX) and B-naphthoflavone (BNF). In untreated cultures the level of the various forms of P-450, determined by immunoblot with the use of specific antibodies, generally declined with time but at markedly different rates. In cultures treated with the inducers decline of the various forms was either unaffected, reduced, or even reversed, so that 96 hr after plating some of these forms appeared to be induced several-fold with respect to the untreated cultures. The forms 2 and 3c were co-induced by PB, RIF or DEX; as in vivo, BNF induced forms 4 and 6. Induction of forms 2, 3c, 4 and 6 was accompanied by stimulation of related monooxygenase activities, benzphetamine demethylase, progesterone 6B hydroxylase and benzpyrene hydroxylase and ethoxyresorufin deethylase, respectively. In all cases, induction was accompanied by an increased rate of de novo synthesis of the protein, determined by radio-immunoprecipitation assay with the use of specific antibodies on [3H]-Leu labeled cell lysate. Both induction and increased de novo synthesis were time- and inducer concentration-dependent. In cultures treated with RIF or BNF de novo synthesis of P-450 3c or of P-450 4 and 6 was correlated with the level of their specific mRNA quantitated from northern blots probed with either pLM3c-4.1 or pLM6.1, two plasmids containing inserted cDNA coding for P-450 3c or P-450 6, respectively. We conclude from these experiments that rabbit hepatocytes in primary monolayer cultures represent suitable models for studying regulation induction and pharmacological implications of the microsomal cytochromes P-450.


Lysosomes and pancreatic islet function. A quantitative estimation of crinophagy in the mouse pancreatic B-cell.
Ultrastructural studies of pancreatic islets have suggested that crinophagy provides a possible mechanism for intracellular degradation of insulin in the insulin-producing B-cells. In the present study, a quantitative estimation of crinophagy in mouse pancreatic islets was attempted by morphometric analysis of lysosomes containing immunoreactive insulin. Isolated islets were incubated in tissue culture for one week in 3.3, 5.5 or 28 mmol/l glucose. The lysosomes of the pancreatic B-cells were identified by morphological and enzyme-cytochemical criteria and divided into three subpopulations comprising primary lysosomes and insulin-positive or insulin-negative secondary lysosomes. Both the volume and numerical density of the primary lysosomes increased with increasing glucose concentration. The proportion of insulin-containing secondary lysosomes was highest at 5.5 and lowest at 3.3 mmol/l glucose. Insulin-negative secondary lysosomes predominated at 3.3 mmol/l glucose. Studies of the dose-response relationships of glucose-stimulated insulin biosynthesis and insulin secretion of the pancreatic islets showed that biosynthesis had an apparent Km-value for glucose of 7.0 mmol/l, whereas it was 14.5 mmol/l for secretion. The pronounced crinophagic activity at 5.5 mmol/l glucose may thus be explained by the difference in glucose sensitivity between insulin biosynthesis and secretion resulting in an intracellular accumulation of insulin-containing secretory granules. The predominance of insulin-negative secondary lysosomes at 3.3 mmol/l glucose may reflect an increased autophagy, whereas the predominance of primary lysosomes at 28 mmol/l glucose may reflect a generally low activity of intracellular degradative processes.


Reproducibility, accuracy and concordance of Accutrend Plus for measuring circulating lipid concentration in adults.
The determination of lipid biomarkers by capillary sampling may be useful in the screening, diagnosis and/or personal management of hyperlipidemia and cardiovascular risk. It remains unclear whether the use of the Accutrend Plus system is appropriate. This study aimed to assess its reproducibility, accuracy and concordance for blood lipid profiling in adults. Fasting capillary total cholesterol (TC) and triglyceride (TG) concentration on Accutrend Plus were compared with their venous analogues obtained by a laboratory reference method in sixty-one adults (27 men and 34 women, aged 33.0 years). Supplementary capillary sampling was performed at two consecutive days taking into account macro-nutrient intake. The day-to-day reproducibility of the Accutrend Plus system proved to be high for TC (ICC = 0.85, P < 0.001), but moderate for TG (ICC = 0.68, P < 0.001). Strong correlations (r > or = 0.80, P < 0.001) with the reference method were found for TC and TG. Mean difference (limits of agreement) were: 0.26 mmol/L (-0.95, 1.47) for TC, and -0.16 mmol/L (-1.29, 0.98) for TG. The concordance for subject classification according to the National Cholesterol Education Program (NCEP) guidelines was significant (P < 0.001), with substantial agreement for TC (K(w) = 0.67), and moderate agreement for TG (K(w) = 0.50). Day-to-day reproducibility of the Accutrend Plus device for TC and TG is not optimal and lacks accuracy when compared to the reference laboratory method. The concordance between both methods for classifying subjects according to the NCEP is inadequate. Accutrend Plus device should not be interchangeably used as a substitution for the standard laboratory methods in the diagnosis of hyperlipidemia.


Vomeronasal inputs to the rodent ventral striatum.
Vertebrates sense chemical signals through the olfactory and vomeronasal systems. In squamate reptiles, which possess the largest vomeronasal system of all vertebrates, the accessory olfactory bulb projects to the nucleus sphericus, which in turn projects to a portion of the ventral striatum known as olfactostriatum. Characteristically, the olfactostriatum is innervated by neuropeptide Y, tyrosine hydroxylase and serotonin immunoreactive fibers. In this study, the possibility that a structure similar to the reptilian olfactostriatum might be present in the mammalian brain has been investigated. Injections of dextran-amines have been aimed at the posteromedial cortical amygdaloid nucleus (the putative mammalian homologue of the reptilian nucleus sphericus) of rats and mice. The resulting anterograde labeling includes the olfactory tubercle, the islands of Calleja and sparse terminal fields in the shell of the nucleus accumbens and ventral pallidum. This projection has been confirmed by injections of retrograde tracers into the ventral striato-pallidum that render retrograde labeling in the posteromedial cortical amygdaloid nucleus. The analysis of the distribution of neuropeptide Y, tyrosine hydroxylase, serotonin and substance P in the ventral striato-pallidum of rats, and the anterograde tracing of the vomeronasal amygdaloid input in the same material confirm that, similar to reptiles, the ventral striatum of mammals includes a specialized vomeronasal structure (olfactory tubercle and islands of Calleja) displaying dense neuropeptide Y-, tyrosine hydroxylase- and serotonin-immunoreactive innervations. The possibility that parts of the accumbens shell and/or ventral pallidum could be included in the mammalian olfactostriatum cannot be discarded.


Atmospheric pollutants in peri-urban forests of Quercus ilex: evidence of pollution abatement and threats for vegetation.
Peri-urban vegetation is generally accepted as a significant remover of atmospheric pollutants, but it could also be threatened by these compounds, with origin in both urban and non-urban areas. To characterize the seasonal and geographical variation of pollutant concentrations and to improve the empirical understanding of the influence of Mediterranean broadleaf evergreen forests on air quality, four forests of Quercus ilex (three peri-urban and one remote) were monitored in different areas in Spain. Concentrations of nitrogen dioxide (NO2), ammonia (NH3), nitric acid (HNO3) and ozone (O3) were measured during 2 years in open areas and inside the forests and aerosols (PM10) were monitored in open areas during 1 year. Ozone was the only air pollutant expected to have direct phytotoxic effects on vegetation according to current thresholds for the protection of vegetation. The concentrations of N compounds were not high enough to directly affect vegetation but could be contributing through atmospheric N deposition to the eutrophization of these ecosystems. Peri-urban forests of Q. ilex showed a significant below-canopy reduction of gaseous concentrations (particularly NH3, with a mean reduction of 29-38%), which indicated the feasibility of these forests to provide an ecosystem service of air quality improvement. Well-designed monitoring programs are needed to further investigate air quality improvement by peri-urban ecosystems while assessing the threat that air pollution can pose to vegetation.


Dietary Quality and Circulating Lipidomic Profiles in 2 Cohorts of Middle-Aged and Older Male Finnish Smokers and American Populations.
Higher dietary quality is associated with lower disease risks and has not been examined extensively with lipidomic profiles. Our goal was to examine associations of the Healthy Eating Index (HEI)-2015, Alternate HEI-2010 (AHEI-2010), and alternate Mediterranean Diet Index (aMED) diet quality indices with serum lipidomic profiles. We conducted a cross-sectional analysis of HEI-2015, AHEI-2010, and aMED with lipidomic profiles from 2 nested case-control studies within the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial (n = 627) and the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study (n = 711). We used multivariable linear regression to determine associations of the indices, derived from baseline food-frequency questionnaires (Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial: 1993-2001, Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study: 1985-1988) with serum concentrations of 904 lipid species and 252 fatty acids (FAs) across 15 lipid classes and 28 total FAs, within each cohort and meta-analyzed results using fixed-effect models for lipids significant at Bonferroni-corrected threshold in common in both cohorts. Adherence to HEI-2015, AHEI-2010, or aMED was associated positively with 31, 41, and 54 lipid species and 8, 6, and 10 class-specific FAs and inversely with 2, 8, and 34 lipid species and 1, 3, and 5 class-specific FAs, respectively. Twenty-five lipid species and 5 class-specific FAs were common to all indices, predominantly triacylglycerols, FA22:6 [docosahexaenoic acid (DHA)]-containing species, and DHA. All indices were positively associated with total FA22:6. AHEI-2010 and aMED were inversely associated with total FA18:1 (oleic acid) and total FA17:0 (margaric acid), respectively. The identified lipids were most associated with components of seafood and plant proteins and unsaturated:saturated fat ratio in HEI-2015; eicosapentaenoic acid plus DHA in AHEI-2010; and fish and monounsaturated:saturated fat ratio in aMED. Adherence to HEI-2015, AHEI-2010, and aMED is associated with serum lipidomic profiles, mostly triacylglycerols or FA22:6-containing species, which are related to seafood and plant proteins, eicosapentaenoic acid-DHA, fish, or fat ratio index components.


Induction of DNA strand breaks and DNA-protein cross-links in normal human skin fibroblasts following exposure to 254 nm UV radiation.
Levels of DNA strand breaks and DNA-protein cross-links (DPCs) were measured using the alkaline elution assay in normal human skin fibroblasts irradiated with 0-200 J m-2 of 254 nm UV radiation and incubated for 0-24 h. On incubation, the yields of both single-strand breaks (SSBs) and DPCs increased with similar kinetics and remained elevated. In addition, when SSBs were measured under conditions in which DPCs were not eliminated by treatment with proteinase K, a measurable yield of SSBs could not be detected. Hence, the SSBs that form in the UV-irradiated cells following incubation appear to be associated with the DPCs.