Circ_0001715 accelerated lung adenocarcinoma process by the miR-1322/CANT1 axis.
Lung adenocarcinoma (LUAD) is a type of lung cancer, which belongs to non-small cell lung cancer and has seriously endangered the physical and mental health of people. The study of circRNAs (circRNAs) has been increasingly hot in recent years, in which circRNAs also play an important regulatory role in cancer. The aim of this study was to investigate the biological molecular mechanisms of circ_0001715 in the progression of LUAD. The expression of circ_0001715, miR-1322 and calcium-activated nucleotidase 1 (CANT1) in LUAD tissues and cell lines was assessed by quantitative reverse transcription PCR (RT-qPCR) and western bot assay. Clone formation assay, 5-Ethynyl-2'-Deoxyuridine (EDU) assay and wound healing assay were used to verify the proliferation ability of cells. Dual-luciferase reporter assay and RNA pull-down assay were performed to characterize the interactions between the three factors. Finally, a mouse tumor model was constructed to assess the tumorigenicity of circ_0001715. RT-qPCR assay results showed that circ_0001715 expression was significantly increased in LUAD tissues and cell lines. Finally, knockdown of circ_0001715 could inhibit tumor growth in vivo. Circ_0001715 regulated the progression of LUAD through the miR-1322/CANT1 axis. The results of this study provided ideas for understanding the molecular mechanisms of circ_0001715 in LUAD.


YouTube: a promotional vehicle for little cigars and cigarillos?
As the third most popular website in the world, messages embedded in the video-sharing site, YouTube, have the potential to influence tobacco-related attitudes, beliefs and behaviours. Despite the growing number of videos depicting little cigars/cigarillos (LCCs), there has been no examination of the portrayal of these products on YouTube. Researchers identified up to the top 20 search results on YouTube by relevance and view count for the following search terms: 'little cigars', 'cigarillos', 'Black and Mild', 'Swisher Sweets', 'White owl', 'Garcia y Vega', and 'Winchester'. Reviewers rated whether videos were 'pro', 'anti' or 'neutral' to the use of LCCs, and documented statistics on the reach and viewer demographics. Several main themes around LCCs were noted, as was video quality (amateur vs professional) and demographics of video participants. Of the 196 videos retrieved, only 56 were unique, eligible videos. The majority of these (n=43) were 'pro' LCCs, 11 were 'neutral', and only two were 'anti' LCCs. Videos were primarily viewed by males in the USA and Canada and most were amateur. Common themes included where to purchase LCCs, their candy flavours, and that they are cheap or cheaper than cigarettes, and 'smooth'. The vast majority of information on YouTube about LCCs promotes their use. It is critical to monitor content on LCCs posted on YouTube, and develop appropriate health messages to counter pro-LCC content, and appropriately inform potential consumers of the harms associated with their use.


Evolution: The rise and fall of island dwarfs and giants.
Islands are arenas for a range of striking evolutionary phenomena, including the island rule - the tendency for larger animals to shrink and smaller animals to enlarge. A new study of insular mammals finds such body size shifts predispose these evolutionary marvels to greater extinction risk.


Epidermal uptake of Pb, Cd, and Zn in tubificid worms.
The epidermal uptake of Pb, Cd, and the essential element Zn by tubificid worms was investigated. The animals were starved and contaminated via the water column at 4°C and 25°C. Atomic absorption spectroscopy (AAS), energy-dispersive X-ray microanalysis (EDX), laser induced mass analysis (LAMMA), electron microscopy, and the sulfide-silver method at the electron microscopical level were used. The sulfidesilver method revealed many reaction products indicating heavy metals in membrane-limited lysosomal structures in epidermal cells of contaminated Limnodrilus udekemianus. In these lysosomes, which were particularly abundant in the epidermis of the hind end, Pb, Cd, and Zn were detected by LAMMA and EDX analysis. Beside this distinct occurrence of the three elements a uniform pattern of reaction products was found in epidermal cells after contamination with Cd and Zn, but not after treatment with Pb. As shown by atomic absorption spectroscopy, Cd was the most enriched element at 25°C, followed by Pb and Zn. Simultaneous application of Zn reduced the uptake of Cd, whereas the uptake of Pb was increased. At 4° C uptake of Pb and Cd was slower than at 25° C, while Zn uptake was blocked completely. This shows that heavy metal uptake can be strongly dependent on environmental conditions, which has to be taken into account if animals are used as monitor organisms for heavy metal pollution in the environment.


Effect of a Behavioral Self-Regulation Intervention on Patient Adherence to Fluid-Intake Restrictions in Hemodialysis: a Randomized Controlled Trial.
The purpose of this study is to evaluate the efficacy of a behavioral self-regulation intervention vs. active control condition using a parallel-group randomized clinical trial with a sample of center hemodialysis patients with chronic kidney disease. Participants were recruited from 8 hemodialysis treatment centers in the Midwest. Eligible patients were (a) fluid nonadherent as defined by an interdialytic weight gain >2.5 kg over a 4-week period, (b) >18 years of age, (c) English-speaking without severe cognitive impairment, (d) treated with center-based hemodialysis for >3 months, and (e) not living in a care facility in which meals were managed. Medical records were used to identify eligible patients. Patients were randomly assigned to either a behavioral self-regulation intervention or active control condition in which groups of 3-8 patients met for hour-long, weekly sessions for 7 weeks at their usual hemodialysis clinic. Primary analyses were intention-to-treat. Sixty-one patients were randomized to the intervention while 58 were assigned to the attention-placebo support and discussion control. Covariate-adjusted between-subjects analyses demonstrated no unique intervention effect for the primary outcome, interdialytic weight gain (β = 0.13, p = 0.48). Significant within-subjects improvement over time was observed for the intervention group (β = -0.32, p = 0.014). The present study found that participation in a behavioral self-regulation intervention resulted in no unique intervention effect on a key indicator of adherence for those with severe chronic kidney disease. There was, however, modest within-subjects improvement in interdialytic weight gain for the intervention group which meshes with other evidence showing the utility of behavioral interventions in this patient population. ClinicalTrials.gov Identifier: NCT01066949.


Forty years of atmospheric radiocarbon monitoring around Bohunice nuclear power plant, Slovakia.
Radiocarbon variations in the atmospheric CO(2) with attenuating amplitudes and decreasing mean values with typical maxima in summer and minima in winter have been observed since 1967 in two localities of Slovakia, in Bratislava and Zlkovce, situated about 60 km NE from Bratislava, only 5 km from the Bohunice Nuclear Power Plant (NPP). The (14)C record in Bratislava has been influenced mainly by fossil CO(2) emissions, in contrast to the Zlkovce record which has been more variable, as it has clearly been affected by operation of the Bohunice NPP. However, during specific meteorological conditions with NE transport of air masses to Bratislava, the effect of the Bohunice NPP has been visible in Bratislava as well. Maximum (14)C concentrations (up to 120% above a natural background) were observed around A1 NPP which used CO(2) with admixture of air as a cooling agent. The (14)C concentrations around four pressurized light water reactors were up to 30% above the background. The Delta(14)C values in the heavily polluted atmosphere of Bratislava were up to 10% and at Zlkovce up to 5% lower than the European clean air represented by the Jungfraujoch Delta(14)C data. Later the Delta(14)C values were similar at both sites, and from 2003 they were close to the European clean air levels. The observed Delta(14)C behaviour in the atmosphere provides a unique evidence of decreased fossil fuel CO(2) emissions in the region, as well as the long-term effect of the Bohunice NPP on the Bratislava and Zlkovce stations. The estimated annual radiation doses to the local public due to digestion of radiocarbon contaminated food have been estimated to be around 3 microSv.


Morphological structure of propagules and electrophoretic karyotype analysis of false smut Villosiclava virens in rice.
The target pathogen Villosiclava virens (teleomorph: claviceps oryzae-sativae) was isolated from the infected rice, where it caused false smut. In our study, the forming processes of the chlamydospores, chlamydospore balls, conidiospores, and secondary conidiospores during the asexual reproduction were observed more precisely and in greater detail than previous descriptions. The microstructure of the infected rice kernel showed that the outer dense chlamydospores piled around the false smut balls grown on XBZ medium; moreover the sclerotia consisting of dense mycelium were found. The different morphology was observed across the different growing conditions. In addition, we observed the nuclear numbers of both the conidiospores and hyphae using 4',6-diamidino-2-phenylindole (DAPI) staining. Because the fungus has small chromosomes and the numbers were not previously known, we analyzed the electrophoretic karyotype using a pulsed field gel electrophoresis (PFGE) technique. The results showed that V. virens has at least 10 chromosomes ranging in size from 0.6 kb to 6 Mb. The V. virens genome size is estimated to be 23 Mb. Here, we report the morphological characteristics of the fungus and the process of asexual spores forming asexual propagules, along with the first analyze the molecular karyotype of V. virens. These results supply a foundation for further study of the pathogenicity and biology of this devastating pathogen.


Multiwavelength method for measuring concentration of free cytosolic calcium using the fluorescent probe indo-1.
It is assumed that the spectra of fluorescent probes indo-1 and fura-2 in the cytoplasm are linear combinations of the spectra of calcium-bound and free probes with weight factors proportional to the concentrations of these forms. When the concentration of calcium is measured by the dual-wavelength method, the above assumption is employed without testing. A multiwavelength method for measuring free cytosolic calcium concentration is described in the present study. The method is based on the registration of the fluorescence spectra of the probe with an optical multichannel analyzer and deconvolution of the spectra into components, corresponding to free and bound forms of the probe. A mismatch is also calculated to allow estimation of deconvolution accuracy. It was found that the spectra, recorded in aqueous calibration solution with varying calcium concentrations, can be deconvoluted into components, obtained both in the absence of calcium and at its saturating concentration. When the spectrum of the probe in the cytoplasm is deconvoluted into the same components the mismatch is higher. When aqueous calibration is used, the cytosolic calcium concentration determined by the dual-wavelength method is dependent considerably on the selected wavelengths. Our data indicate that this phenomenon may be associated with the lower polarity of cytoplasm compared to the aqueous calibration solution. Addition of either ethanol or glycerol into the calibration medium results in a considerable decrease in the mismatch. The optimal concentration of ethanol is 22-32%, and depends on the type and condition of cells tested. It is shown that the use of calibration spectra obtained in aqueous solutions leads to considerable overestimation of cytosolic calcium concentration.


New cochlear implant technologies improve performance in post-meningitic deaf patients.
The objective of the study was to compare the performance of cochlear implantation between post-meningitic and non-meningitic patients, and to evaluate the impact on hearing outcome of technical advances in cochlear implant technology. Retrospective chart review was used as the study design. Twenty adults with post-meningitic profound hearing loss receiving unilateral or bilateral cochlear implants between 1990 and 2008 were tested. Results were compared to a control group of 46 adults implanted for a non-meningitic hearing loss, with the same pre-operative speech scores. Speech scores were poorer in post-meningitic patients compared to those of control group, whatever the duration after implantation (p < 0.0001). Speech scores of subjects implanted and fitted before 2001 were compared to those of subjects implanted after 2001, with the same duration of hearing loss. Performance improved with implants and processors available after 2001, with a magnitude of improvement higher in post-meningitic patients (p < 0.0001 and p < 0.05 in post-meningitic and control groups, respectively, two-way ANOVA). Consequently, speech scores of post-meningitic patients implanted after 2001 achieved those of control subjects (two-way ANOVA). Advances in cochlear implant technology and coding strategy improve hearing outcome in post-meningitic adult patients, who now achieve similar performance as those of non-meningitic patients.


Scaphotrapezial trapezoidal arthrosis.
Scaphotrapezial trapezoidal degenerative arthritis as an isolated entity or as a preponderant part of a pantrapezial degenerative process was seen in 49 hands of 34 patients. Pain and weakness were the primary complaints. Point tenderness was present in all but two hands. Significant loss of grasp and pinch strength was noted in 18 hands, and loss of wrist motion occurred in 16 hands. Roentgenographic findings included narrowing of the scaphotrapezial trapezoidal joint space, subarticular cortical sclerosis, and cyst formation. A dorsiflexed position of the scaphoid, with a scapholunate angle of less than 45 degrees, was noted in 38 of the 45 hands. Late collapse of the intercarpal joint with dorsiflexion instability of the lunate also was noted. Twenty-nine hands were treated conservatively by protective splints, corticosteroid injections, and avoidance of exacerbating activities. Surgical treatments consisted of fibrous arthroplasty (four), silicone interposition arthroplasty (six), trapezial replacement arthroplasty (three), arthrodesis (five), joint debridement (one), and trapezial excision (one). Late results were more encouraging than were early results and were moderately satisfactory for each method. Improvement in grasp and pinch strength was most predictable for arthrodesis, although wrist motion was diminished.


Correlation of hepatitis B surface antigen level with response to telbivudine in naive patients with chronic hepatitis B.
Hepatitis B surface antigen (HBsAg) has become a marker to judge immunological response to hepatitis B therapy. Quantified serum HBsAg levels can predict the response to pegylated interferon and entecavir. In this study, we aimed to explore the correlation of serum HBsAg levels with response to telbivudine (LdT) treatment in patients with chronic hepatitis B (CHB). Seventy-three treatment-naive CHB patients were recruited and received LdT monotherapy for 52 weeks and serial HBsAg levels were measured at five protocol time points. According to therapeutic efficacy at week 52, three subgroups of patients were identified, including complete responders (CR), partial responders (PR) and non-responders (NR). After 52 weeks of treatment, CR, PR and NR represented 19 (26%), 33 (45%) and 21 (29%) patients in the sample of 73, respectively. The median values of baseline HBsAg (log10 IU/mL) were 4.05, 4.50 and 5.03 for CR, PR and NR, respectively. There was a distinct decline of HBsAg at week 52; median log10 HBsAg levels (IU/mL) were 3.61 (CR), 3.86 (PR) and 4.31 (NR). Positive correlation between HBsAg levels and HBV DNA loads was observed in the group of NR and early antiviral treatment of PR, but not in CR. Initial HBsAg level was closely correlated with the efficacy of LdT. Patients with low HBsAg levels presented satisfactory responses. Therefore, initial level and correlation with HBV DNA of the serum HBsAg levels could predict responsiveness in CHB patients receiving LdT.


Prediction of potential molecular markers of bovine mastitis by meta-analysis of differentially expressed genes using combined p value and robust rank aggregation.
Bovine mastitis causes significant economic loss to the dairy industry by affecting milk quality and quantity. Escherichia coli and Staphylococcus aureus are the two common mastitis-causing bacteria among the consortia of mastitis pathogens, wherein E. coli is an opportunistic environmental pathogen, and S. aureus is a contagious pathogen. This study was designed to predict molecular markers of bovine mastitis by meta-analysis of differentially expressed genes (DEG) in E. coli- or S. aureus-infected mammary epithelial cells (MECs) using p value combination and robust rank aggregation (RRA) methods. High-throughput transcriptome of bovine MECs, infected with E. coli or S. aureus, were analyzed, and correlation of z-scores were computed for the expression datasets to identify the lineage profile and functional ontology of DEGs. Key pathways enriched in infected MECs were deciphered by Gene Set Enrichment Analysis (GSEA), following which combined p value and RRA were used to perform DEG meta-analysis to limit type I error in the analysis. The miRNA-gene networks were then built to uncover potential molecular markers of mastitis. Lineage profiling of MECs showed that the gene expression levels were associated with mammary tissue lineage. The up-regulated genes were enriched in immune-related pathways, whereas down-regulated genes influenced the cellular processes. GSEA analysis of DEGs deciphered the involvement of Toll-like receptor (TLR), and NF-kappa B signaling pathway during infection. Comparison after meta-analysis yielded with genes ZC3H12A, RND1, and MAP3K8 having significant expression levels in both E. coli and S. aureus dataset, and on evaluating miRNA-gene network, 7 pairs were common to both sets identifying them as potential molecular markers.


Excitable chemical reaction systems. II. Several pulses on the ring fiber.
The parameters of the Field, Noyes and Körös model for the Belousov-Zhabotinskii reaction were chosen in such a way that the given system is excitable. It is shown by simulation that several pulses on the ring fiber, contrary to experience with known excitable media, may arrange themselves in a non-symmetrical manner while the equilibrium distances are stable. The condition for such behavior is the non-monotonic dependence of the propagation velocity, v, on the time, T, elpased since the previous excitation at that point.


Flecainide. A preliminary review of its pharmacodynamic properties and therapeutic efficacy.
Flecainide is a Class I antiarrhythmic drug of the local anaesthetic type. It can be given either intravenously or orally and its pharmacokinetic properties allow relatively long (12 hours) dosing intervals with oral administration. In several open and a few controlled therapeutic trials, orally administered flecainide has brought about a greater than 90% suppression of ventricular ectopic beats in about 80% of patients. A similar percentage of patients (83%) experienced at least an 80% suppression of their ventricular tachycardia in these trials. A slightly greater response rate was reported with intravenous infusion of flecainide. Initial results in arrhythmias complicating the Wolff-Parkinson-White syndrome have been favourable. Comparative trials are few in number but flecainide has proved to be more effective than quinidine, and possibly more effective than disopyramide, mexiletine, tocainide and propafenone, in suppressing ventricular ectopic activity. The most commonly reported extracardiac adverse effects have been dizziness and visual disturbances. Proarrhythmic effects have been reported in 7 to 8% of patients, with a higher incidence in patients with serious ventricular tachycardia and reduced myocardial function. The moderate negative inotropic effects of flecainide can become clinically significant in patients with impaired ventricular function. Thus flecainide, with its convenient dose schedule and apparently low incidence of serious side effects, would appear to be a useful addition to the antiarrhythmic agents available. Further studies are needed though, to confirm its long term tolerability when used prophylactically.


Children's Artwork: Its Value in Psychotherapy in Pediatric Palliative Care.
Pediatric palliative care is a comprehensive treatment approach (physical, psychological, social, spiritual) for children living with life-threatening conditions. These patients and siblings, as well as children of ill parents, face extraordinary psychological challenges. Structured art techniques incorporated into psychotherapy can be powerful for children dealing with life-and-death realities. This article provides the rationale, instructions, and examples for 3 techniques that the author has adapted for children facing illness and bereavement. Although these art techniques are simple to administer, they frequently evoke complex and powerful responses and thus are intended for use by or in consultation with mental health professionals.


Clinical features of Talaromyces marneffei infection in HIV-positive and HIV-negative individuals: A retrospective study in southern China.
Talaromyces marneffei (TSM) is a temperature-dependent dimorphic fungus endemic to Southeast Asia and southern China. As the number of people at risk of TSM infection continues to increase, the clinical manifestations are becoming increasingly complex, posing challenges for clinical management. In this study, we analyzed the medical records of 99 patients (71 human immunodeficiency virus [HIV]-positive and 28 HIV-negative) diagnosed with TSM infection from January 1, 2017, to December 31, 2022, in southern China and compared the clinical manifestations in HIV-positive and HIV-negative patients. Most patients (83/99, 84%) were male. The incidence of skin and soft tissue involvement (48% vs. 21%, P = .016); disseminated infection with blood circulation, hematopoietic, lymphatic, alimentary, or central nervous system involvement (69% vs. 36%, P = .002); and gastrointestinal bleeding (33% vs. 9%, P = .023) was higher in the HIV-positive group than the HIV-negative group. The HIV-positive group also had significantly higher alanine aminotransferase (ALT) levels (31 [26-42] vs. 14 [11-16] U/l, P < .001) and ALT/aspartate transaminase ratio (1.9 [1.5-2.2] vs. 1.3 [1.1-1.6], P = .006) than the HIV-negative group. The time to diagnosis (5.5 ± 1.1 vs. 5.1 ± 1.4 days, P = .103), antifungal regimen (P = .278), case fatality rate (20% vs. 21%, P = .849), and relapse/reinfection rate (11% vs. 19%, P = .576) did not differ significantly between the HIV-positive and HIV-negative groups. Poor antiretroviral therapy adherence (OR = 26.19, 95%CI 3.26-210.70, P = .002), advanced age (OR = 1.13, 95%CI 1.03-1.23, P = .010), and Epstein-Barr virus co-infection (OR = 37.13, 95%CI 3.03-455.64, P = .005) were independent risk factors for all-cause mortality from TSM infection in HIV-positive patients. Overall, the predominant infection sites, clinical manifestations, and complications of TSM infection differed by HIV status. However, with prompt diagnosis and appropriate treatment, HIV-positive patients with TSM infection can have similar outcomes to HIV-negative patients.


Initial experience with video-assisted thoracoscopic lobectomy.
In the past 2 years, the development of video-assisted thoracic surgery (VATS) has allowed thoracoscopy to rapidly evolve from a primarily diagnostic tool to a therapeutic tool with potentially useful applications. As with laparoscopy, the initial enthusiasm for VATS must be tempered by the reality of technical inadequacies, complications, and results that have yet to withstand careful scientific scrutiny. The purpose of this study was to determine the technical feasibility of a VATS lobectomy. Forty-four patients with primary bronchogenic carcinoma were evaluated and accepted as potential candidates for VATS lobectomy. After complete preoperative staging, these patients were in clinical stage I. All patients had normal arterial blood gases and adequate pulmonary function (forced expiratory volume in 1 second > 1.5 L) to tolerate a lobectomy. At the time of operation, 3 patients were found to have N2 disease and were excluded from the study. In 35 of the remaining 41 patients, a successful VATS lobectomy was accomplished through two thoracoscopy ports and a non-rib-spreading 6- to 8-cm "access" thoracotomy. There were no major intraoperative complications that necessitated conversion to an open thoracotomy. Mean operative time was 153 +/- 26 minutes. All 35 patients recovered uneventfully with a mean hospital stay of 5.7 +/- 1.6 days. A VATS lobectomy is technically feasible using our approach and is potentially safe. However, major advances in thoracoscopy imaging and instrumentation are necessary before this procedure will have the potential for widespread acceptance. Also, the advantages of VATS lobectomy over accepted surgical techniques will have to be carefully documented in randomized trials.


Adsorption and desorption of DNA tuned by hydroxyl groups in graphite oxides-based solid extraction material.
The extraction of DNA is the most crucial method used in molecular biology. Up to date silica matrices has been widely applied as solid support for selective DNA adsorption and extraction. However, since adsorption force of SiOH functional groups is much greater than that of desorption force, the DNA extraction efficiency of silica surfaces is limited. In order to increase the DNA extraction yield, a new surface with different functional groups which possess of greater desorption property is required. In this study, we proposed cellulose/graphite oxide (GO) composite as an alternative material for DNA adsorption and extraction. GO/Cellulose composite provides the major adsorption and desorption of DNA by COH, which belongs to alkyl or phenol type of OH functional group. Compared to SiOH, COH is less polarized and reactive, therefore the composite might provide a higher desorption of DNA during the elution process. The GO/cellulose composite were prepared in spherical structure by mixing urea, cellulose, NaOH, Graphite oxide and water. The concentration of GO within the composites were controlled to be 0-4.15 wt.%. The extraction yield of DNA increased with increasing weight percentage of GO. The highest yield was achieved at 4.15 wt.% GO, where the extraction efficiency was reported as 660.4 ng/μl when applying 2M GuHCl as the binding buffer. The absorbance ratios between 260 nm and 280 nm (A260/A280) of the DNA elution was demonstrated as 1.86, indicating the extracted DNA consisted of high purity. The results proved that GO/cellulose composite provides a simple method for selective DNA extraction with high extraction efficiency of pure DNA.


Insecticide drift deposition on noncrop plant surfaces and its impact on two beneficial nontarget arthropods, Aphidius colemani Viereck (Hymenoptera, Braconidae) and Coccinella septempunctata L. (Coleoptera, Coccinellidae).
Drift of the pyrethroid insecticide lambda-cyhalothrin into field margins bordering wheat and its effect on nontarget arthropods was investigated under field conditions. Insecticide deposition on leaf surfaces of broad beans exposed within field margins at 1-, 2-, and 3-m distance from the field edge and directly within the crop was quantified using a fluorescent tracer. The toxicity of leaf deposits to the aphid parasitoid Aphidius colemani Viereck (Hymenoptera: Braconidae) and to larvae of the ladybird Coccinella septempunctata L. (Coleoptera: Coccinellidae) was estimated. Mean corrected mortalities of A. colemani were < 50% (1 m) and < 30% (2 and 3 m), and mortalities of C. septempunctata larvae were < or = 80% (1 m), < or = 67% (2 m), and < or = 52% (3 m) 12 and 24 h postexposure to drift deposits. High positive correlation coefficients proved the relationship between leaf deposits and mortalities of test organisms. The connection of drift deposit measurement and exposure bioassay provides a promising approach to determining effects of insecticide drift on nontarget arthropods. However, due to the patchiness of drift deposition on noncrop plant surfaces, risks to nontarget arthropods at defined distances from the sprayed crop are not highly predictable.


Disruption of adaptor protein 2μ (AP-2μ) in cochlear hair cells impairs vesicle reloading of synaptic release sites and hearing.
Active zones (AZs) of inner hair cells (IHCs) indefatigably release hundreds of vesicles per second, requiring each release site to reload vesicles at tens per second. Here, we report that the endocytic adaptor protein 2μ (AP-2μ) is required for release site replenishment and hearing. We show that hair cell-specific disruption of AP-2μ slows IHC exocytosis immediately after fusion of the readily releasable pool of vesicles, despite normal abundance of membrane-proximal vesicles and intact endocytic membrane retrieval. Sound-driven postsynaptic spiking was reduced in a use-dependent manner, and the altered interspike interval statistics suggested a slowed reloading of release sites. Sustained strong stimulation led to accumulation of endosome-like vacuoles, fewer clathrin-coated endocytic intermediates, and vesicle depletion of the membrane-distal synaptic ribbon in AP-2μ-deficient IHCs, indicating a further role of AP-2μ in clathrin-dependent vesicle reformation on a timescale of many seconds. Finally, we show that AP-2 sorts its IHC-cargo otoferlin. We propose that binding of AP-2 to otoferlin facilitates replenishment of release sites, for example, via speeding AZ clearance of exocytosed material, in addition to a role of AP-2 in synaptic vesicle reformation.


Thinking Outside the Lungs: A Case of Disseminated Abdominal Tuberculosis.
BACKGROUND Extrapulmonary tuberculosis (TB) occurs in up to one-fifth of all cases of TB, with abdominal TB accounting for 5% of all cases. It is an uncommon diagnosis in the Western world, where it is primarily identified in immigrant and immunocompromised populations. CASE REPORT We review a case in which a 47-year-old Nepalese woman with a history of cognitive dysfunction secondary to epilepsy presented with decreased appetite and diffuse abdominal pain. She was hypoxic and febrile on initial exam, and imaging indicated lung consolidation, right-sided pleural effusion, and thickening and nodularity of the omentum with patchy wall thickening of the colon. After failing to improve on a standard antibiotic regimen for treatment of pneumonia and colitis, the differential was broadened to include TB. Interferon-g release assay was subsequently found to be positive, and omental and peritoneal biopsies were obtained. The patient was started on an empiric course of rifampin, isoniazid, ethambutol, pyrazinamide, and pyridoxine. Laboratory testing revealed no immunochemical evidence of Mycobacterium species, however, Ziehl-Neelsen acid-fast stain was positive with rare acid-fast bacilli identified. CONCLUSIONS Peritoneal TB carries significant morbidity and mortality if undiagnosed or untreated. Diagnosis is challenging in the absence of a single test that can confirm or exclude this condition. In combination with clinical suspicion, it is crucial to explore history regarding socio-epidemiology (travel, incarceration, occupation, homelessness, sick contacts) and immunological risk (drug use, chemotherapy) in patients with constitutional symptoms.


[Reoperative treatment of recurrent retroperitoneal tumor].
To improve the resection rate and survival rate for recurrent retroperitoneal tumor (RRT). We analysed 34 patients with RRT who were treated in our hospital from 1987 to 1995. One patient had benign tumor, the others malignant. Because 7 patients were operated on over one time, we performed 53 operations including complete resection in 42 patients, palliative resection 8 and biopsy 3. The 1-and 2-year survival rates for complete resection cases were 71.2% and 65.3%. The patients who received palliative resection and biopsy died within one year. The diagnosis, preoperative preparation and intraoperative management of RRT are important to operative safety and survival rate. We emphasize the importance of reoperation and complete resection for RRT.


Ethanol inhibits the peak of muscarinic receptor-stimulated formation of inositol 1,4,5-trisphosphate in neuroblastoma SH-SY5Y cells.
The effect of ethanol on muscarinic receptor-stimulated formation of inositol 1,4,5-trisphosphate was studied in human neuroblastoma SH-SY5Y cells. Stimulation with carbachol induced a biphasic increase of inositol 1,4,5-triphosphate with an initial peak after 10 sec declining to a plateau phase of elevation above basal levels, which was sustained for at least 5 min in the presence of agonist. The peak, but not the plateau phase, was concentration-dependently decreased by exposure to ethanol. Maximal inhibition was obtained within 30 sec of exposure to ethanol. Ethanol caused an increase in the EC50 value of carbachol for the initial rate of inositol 1,4,5-trisphosphate formation, measured after 10 sec of stimulation, from 98 microM in the absence to 196 microM in the presence of 100 mM ethanol. The potencies of pirenzepine and hexahydro-sila-difenidol hydrochloride for inhibiting [3H]quinuclidinyl benzilate binding and inositol 1,4,5-trisphosphate formation suggest that both phases are mediated via the muscarinic M1 receptor. Phorbol 12-myristate 13-acetate inhibited both phases of inositol 1,4,5-trisphosphate formation, whereas okadaic acid and modulators of cAMP-dependent protein kinase were without any effect. There was no inhibitory effect of ethanol when protein kinase C was inhibited by H7 and calphostin C, indicating that the ethanol effect is dependent on protein kinase C activity.


The associations between interleukin-1 polymorphisms and susceptibility to ankylosing spondylitis: a meta-analysis.
The aim of this study was to determine whether polymorphisms of interleukin-1 (IL-1) confer susceptibility to ankylosing spondylitis (AS). The authors conducted meta-analyses on associations between IL-1 polymorphisms and AS susceptibility, using fixed or random effects models. In order to avoid duplications and data previously subjected to meta-analysis, we performed meta-analysis on studies if new data on IL-1 polymorphisms in AS were reported. A total of nine studies consisting of 20 separate comparisons of association between IL-1 polymorphisms and AS susceptibility were included in this meta-analysis. These were performed on European, Asian, and Latin American population samples. Meta-analysis revealed a significant association between the 2 allele of the IL-1F10.3 polymorphism (rs3811581) and the risk of developing AS in Europeans (OR=0.775, 95% CI=0.605-0.992, P=0.043). Furthermore, the OR of the 2 allele of IL-1A+889 (rs1800587) was found to be significantly increased in Europeans with AS (OR=1.357, 95% CI=1.085-1.697, P=0.007). However, meta-analyses of the IL-1B-511, IL-1B+3953, and ILF7.1 polymorphisms and of the variable numbers of tandem repeats of the IL-1 receptor antagonist gene (IL-1RN VNTR) revealed no association between AS and these polymorphisms. In addition to the three known IL-1 polymorphisms, rs2856836, rs17561, and rs1894399, found in previous meta-analysis, this meta-analysis shows that the IL-1F10.3 and IL-1A+889 polymorphisms are associated with the development of AS in Europeans but not in Asians.


The relationship between safety-net hospital status and outcomes among elderly head and neck cancer patients.
The impact of safety-net status, case volume, and outcomes among geriatric head and neck cancer patients is unknown. Chi-square tests and Student's t tests to compare head and neck surgery outcomes of elderly patients between safety-net and non-safety-net hospitals. Multivariable linear regressions to determine predictors of outcome variables including mortality index, ICU stays, 30-day readmission, total direct cost, and direct cost index. Compared with non-safety-net hospitals, safety-net hospitals had a higher average mortality index (1.04 vs. 0.32, p = 0.001), higher mortality rate (1% vs. 0.5%, p = 0.002), and higher direct cost index (p = 0.001). A multivariable model of mortality index found the interaction between safety-net status and medium case volume was predictive of higher mortality index (p = 0.006). Safety-net status is correlated with higher mortality index and cost in geriatric head and neck cancer patients. The interaction between medium volume and safety-net status is independently predictive of higher mortality index.


TLR4/CD14-mediated PI3K activation is an essential component of interferon-dependent VSV resistance in macrophages.
Phosphatidylinositol-3-phosphate kinase (PI3K) has been reported to exhibit anti-inflammatory roles as a negative modulator of the NF-kappaB pathway (MyD88- and Mal-dependent) triggered upon Toll-like receptor (TLR)4 activation by lipopolysaccharide (LPS). Here, we investigated the role of PI3K on the TLR4-dependent, MyD88-independent signaling cascade which is activated in macrophages infected by Vesicular Stomatitis Virus (VSV) and leads to interferon production, thus conferring antiviral protection. We show that VSV induces TLR4 (and CD14)-dependent Akt phosphorylation. We observed hypersusceptibility to viral infections after pharmacological inactivation of the PI3K pathway in macrophages, which indicates that normal PI3K functions are critical for type I interferon synthesis and viral resistance. Conversely, we noticed increased resistance in macrophages isolated from genetically modified mice in which the PI3K pathway is constitutively active. Our data, which demonstrate that PI3K-Akt axis is an important component of the TLR4-dependent antiviral mechanism, also indicate that pharmacological modulation of this pathway to regulate the inflammatory response could promote viral susceptibility.


A bone-conserving revision stem for unstable intertrochanteric fractures of the geriatric osteoporotic population.
Primary hemiarthroplasty is gaining popularity for the treatment of unstable intertrochanteric fractures in geriatric patients with severe osteoporosis. This study evaluated early clinical and radiographic outcomes by using a bone-conserving revision stem for unstable intertrochanteric fractures in the geriatric osteoporotic population. A retrospective study involving 31 patients with unstable intertrochanteric fractures was conducted. The patients were aged 82.1 years on average. All patients underwent primary hemiarthroplasty using bone-conserving, fully porous-coated revision stem. The operative time, intraoperative blood loss, length of hospitalization, and need for blood transfusion were noted during the hospital stay. Postoperative complications, including dislocations, deep venous thrombosis, infections, peri-prosthetic fractures, and frontal thigh pain were also recorded. Koval's category was used to quantify activity level, and Harris hip score (HHS) was used for functional assessment. Radiographic outcomes, including osteolysis, bone ingrowth, subsidence of the femoral component, lower limb length discrepancy, and heterotopic ossification, were collected at each follow-up. The 31 patients were followed for an average time of 23 months postoperatively. The average operative time lasted for 74.2 min, while the mean intraoperative blood loss was 200.1 ml, with an average hemoglobin decrease of 11.1 g/L after the procedure. The mean visual analog scale (VAS) score for pain dropped from 7.4 preoperatively to 2.4 at the 4-week follow-up. At the latest follow-up, the mean Harris hip score was 82.1, and the VAS was 1.7. No intraoperative or postoperative peri-prosthetic fractures were noted. Postoperative complications included one case of thrombosis formation in the posterior tibial vein and one case of congestive heart failure. Both patients were discharged uneventfully after treatment. Radiographically, none of the hips had evidence of stem loosening or osteolysis. Within the follow-up period of 23 months, the mortality rate was 3.2% (1/31), and no revision surgeries were required. Primary hemiarthroplasty using a bone-conserving, cementless revision stem could serve as a reliable alternative for the treatment of unstable intertrochanteric fractures in the geriatric population with osteoporosis.


Laparoscopic extraction of a swallowed fork in a patient first diagnosed with bulimia nervosa.
Swallowed foreign bodies are relatively common problem. The first reports date back about 3000 years. The first medical report was done by Mestivier in 1759. Several studies show that up to 90% of the foreign bodies (FBs) pass spontaneously and 10% to 20% require an endoscopic removal. Surgical intervention is only indicated in approximately 1% of the cases. Surgical intervention is only necessary, for example, when large or sharp FBs are involved because of the potential risk of perforation and obstruction. The surgical therapy can be carried out by means of laparotomy or laparoscopy; laparoscopy has to be given the first priority whenever possible. The advantages of a laparoscopic procedure are well-known: reduced postoperative pain, better lung function, less postoperative bowel obstruction, shorter hospital stay, and faster reconvalescence. We report an unusual case of an unintentional ingested fork, which required a laparoscopic extraction using three trocars. The 20-year-old female patient was then subsequently diagnosed with bulimia nervosa for the first time during her hospital stay. The patient was discharged home on the fourth postoperative day after an unremarkable course.


Childhood thyroid cancer in England and Wales.
A total of 154 cases of thyroid cancer in children under 15 were registered in England and Wales over a period of 30 years, an incidence of about 0.5 per million per year. A total of 4.5 cases per year were registered in 1963-72, 4.9 in 1973-82 and 5.8 in 1983-92. A rapid rise in incidence with age occurred after the age of 5. Malignancy was confirmed in 92% of the cases in which tissue was available. Of these, 68% were papillary carcinomas, 11% follicular carcinomas and 17% medullary carcinomas. There were two spindle cell tumours with mucous cysts and one teratoma. The increased frequency but small size of medullary carcinomas in the second half of the period suggested that this increase was due to the introduction of screening; it accounted for most of the rise in crude incidence rats with time. The sex ratio (F:M) in all registered cases in the differentiated follicular cell carcinoma groups in children aged under 10 was 1.2:1, and 3.6:1 in the older children. Five children with differentiated thyroid cancer of follicular cell origin died up to 17 years after diagnosis. Two of the eight children aged 9 or less with a 20 year follow-up died, compared with three of 28 older children. An unusual group of differentiated carcinomas showed solid or follicular architecture. These tumours were unencapsulated, often widely invasive, contained psammoma bodies but little or no papillary architecture and the nuclei often lacked prominent grooving. This childhood type of papillary carcinoma contrasted with the classical type commonly found in the adult, which was present in one of 13 confirmed papillary carcinomas in children aged less than 10, compared with 20 of 35 older children. These observations show that thyroid carcinoma in very young children has a different spectrum of histological types from both older children and adults. From the age of about 10 well-differentiated papillary carcinomas rapidly increase in frequency in females, so that the other types come to form only a small proportion of the total. These differences, and the lower incidence but poorer prognosis of thyroid carcinoma in men and the poorer prognosis in post- as compared with premenopausal women, are compatible with a major role for sex hormones in thyroid carcinogenesis in females during the reproductive period. This study documents the incidence of childhood thyroid cancer in England and Wales, explains the rise in crude incidence rates, shows differences between carcinomas in children under and over the age of ten which may correlate with puberty, and draws attention to an unusual aggressive type of childhood papillary carcinoma. It illustrates the value to crude registry data of a pathology review.


Failure of intravenous fluid therapies to decrease serum sodium levels in elderly hospitalized patients.
Elderly patients may have a tendency to develop hyponatremia due to sensitivity to stimuli that release ADH as well as an impaired ability to excrete a water load. We evaluated changes in serum sodium in elderly hospitalized patients who received various forms of intravenous fluid therapies. All patients were required to have a baseline serum sodium of 136-145 meq/L. Fourteen patients were enrolled in the study. The mean age was 82.9 +/- 6.8 years (mean +/- SEM). Thirty-six % were nursing home residents. Seventy-nine % were females. Seventy-two % received half normal saline and the remainder received normal saline as intravenous fluid therapy. The patients received a mean of 1098 +/- 145 mL of intravenous fluid per day, in addition to oral fluids. Mean follow up period was 5.9 days (3-10 days). Mean baseline serum sodium was 140.2 +/- 0.7 meq/L andmean follow up serum sodium was 141.4 +/- 0.9 meq/L. The m ean baseline BUN was 25 +/- 3.6 mg/dL and mean follow u BUN was 19.6 +/- 3.4 mg/dL. The mean baseline serum creatinine was 0.9 +/- 0.1 mg/dL and mean follow up creatinine was 0.9 +/- 0.1 mg/dL. The postintravenous fluid therapy serum sodium in the group receiving half normal saline was 141.7 +/- 0.7 meq/L and 140.8 +/- 3 meq/L in the normal saline group. No significant difference was observed between the pre and post fluid therapy for any of these paramenters (p > 0.05). Mean baseline plasma renin activity was 1.6 +/- 0.7 ng/ml/hour and fifty-seven % had PRA of less than 1 ng/ml/hour. Mean plasma aldosterone was 8.5 +/- 1.8 ng/mL and forty-two % were less than 5.5 ng/mL. Plasma ADH and ANP was 5.7 +/- 3.4 pg/mL and 83.6 +/- 26.9 pg/mL, respectively. Mean serum and urine osmolalities were 290 +/- 3.1 mOsm/kg and 471 +/- 57.7 mOsm/kg, respectively. No patient developed hyponatremia and 7 of the 14 patients experienced an increase in serum sodium during the follow up period. We conclude that many elderly patients hospitalized for acute medical illnesses either maintain a stable serum sodium or experience an increase in serum sodium. This occurs because total fluids administered to these patients are generally insufficient.


Functional characterisation of mutations in the ligand-binding domain of the androgen receptor gene in patients with androgen insensitivity syndrome.
Five mutations in the ligand-binding domain of the androgen receptor gene were identified in patients with complete (A765T, C784Y, R831X and M895T) or partial (R840G) androgen insensitivity. A765T and R831X have been reported previously whereas the other three mutations are novel. Receptors carrying these mutations were transiently expressed in COS-1 cells, and androgen binding and capacity to transactivate an androgen-responsive reporter gene were assayed. C784Y led to abolished androgen binding and transactivating capacity, R840G and M895T showed reduced specific binding and partial transactivation. The in vitro functions of the R840G and M895T mutants were improved with supraphysiological concentrations of steroid.


Normal aging in the central nervous system: quantitative MR diffusion-tensor analysis.
The purpose of this study is to elucidate changes in mean diffusivity (ADC) and fractional anisotropy (FA) using MR diffusion tensor imaging (DTI) in the central nervous system during normal aging. We studied 50 normal volunteers (30 men, 20 women; mean age 44.8 +/- 14.0; age range, 21-69 years) without disorders affecting the central nervous system. The frontal, parietal white matter, lentiform nucleus, posterior limb of internal capsule, thalamus, genu and splenium of the corpus callosum were selected for investigation. There was no significant difference in ADC or FA between male and female or between the right and left hemisphere. A significant ADC increase with advancing age was observed in frontal white matter (P = 0.010) and lentiform nucleus (P = 0.022). A significant FA decline was found only in the genu of the corpus callosum (P < 0.001) with advancing age. Quantitative diffusion tensor analysis correlate with normal aging and may help in assessing normal age-related changes and serve as a standard for comparison with neurodegenerative disorders.


Power ultrasound in the meat industry (freezing, cooking and fermentation): Mechanisms, advances and challenges.
High intensity ultrasound (HIUS) has a wide range of applications in different sectors of food processing. It is a promising and emerging technology demonstrating the potential to promote food processes without or at least damage to the quality of products. Among the processes of the meat industry, freezing, thawing, cooking and fermentation are very sensitive and important, because they have significant effects on product quality and are also very energy and time consuming. This review paper provides an interpretation of high intensity ultrasound (HIUS) applications, a summary of recent outstanding published research and an overview of the freezing/thawing, cooking/frying and fermentation processes in meat and its products assisted by HIUS. The effects, benefits and drawbacks as well as the challenges ahead in the commercialization of this technology in the meat industry are studied. The research results confirmed that the use of HIUS in the meat freezing/thawing, cooking/frying and fermentation in combination with the corresponding processing methods demonstrates a great potential to promote the process, improve the general quality of the final product and reduce the time and energy required. However, many issues remain that require further research to address these challenges. These challenges and subsequent research that is useful for developing and increasing the efficiency of this technology have been reviewed. After the literature review, it is concluded that HIUS may be a useful technology for meat processing because of its significant effects on the quality factors and related process variables that leads to the preservation of the initial nutritional and sensory properties of meat and its products. Of course, research must be continued to eliminate the disadvantages or minimize the undesirable effects of this technology on the final product and to remove barriers to commercialization and optimization of this method.


Maternal and fetal serum levels of total cortisol and cortisone, unbound cortisol, and corticosteroid-binding globulin in vaginal delivery and cesarean section.
The levels of corticosteroid-binding globulin (CBG, transcortin), total cortisol, and unbound cortisol were measured in maternal and fetal serum at the time of cesarean section or vaginal delivery. While the total cortisol levels in maternal serum do not differ significantly in vaginal delivery as compared to cesarean section, cord serum cortisol levels are significantly higher after vaginal delivery. There are no differences associated with the route of delivery between the mean levels of CBG in cord and maternal blood. However, after the vaginal delivery the unbound cortisol level in maternal serum is more than twice as high as that at cesarean section and in cord serum it is seven times as high. These changes reflect changes in total cortisol and may represent a prelabor surge of fetal cortisol production, or a stress response associated with labor, or a combination of both.


Interaction between retinol intake and ISX rs5755368 polymorphism in colorectal cancer risk: a case-control study in a Korean population.
This study aimed to examine whether the ISX rs5755368 genotypes are associated with the effect of dietary retinol consumption on CRC risk. We recruited 923 CRC patients and 1846 controls to identify the association between dietary retinol and CRC risk. Dietary retinol intake was assessed using a semiquantitative food frequency questionnaire. Genotype data were available for 1419 patients (600 cases and 819 controls) of the total study population. Genotyping was performed using an Illumina MEGA Expanded Array. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated using unconditional logistic regression models. Retinol intake was inversely associated with CRC (OR = 0.49; 95% CI = 0.37-0.63). Participants with AA genotype showed lower CRC risk than subjects carrying the G allele (AG + GG) (OR = 0.76; 95% CI = 0.58-0.99). A 68% reduced risk of CRC was related to subjects who had the highest retinol intake and carrying AA genotype compared to the risk of participants consumed the lowest retinol intake and carrying the G allele (OR = 0.32; 95% CI = 0.20-0.53; P interaction = 0.026). Retinol intake could be a protective factor for CRC risk while this association could be strengthened among individuals carrying the homozygous AA genotype.


Passive Acoustic Monitoring of the Environmental Impact of Oil Exploration on Marine Mammals in the Gulf of Mexico.
The Gulf of Mexico is a region densely populated by marine mammals that must adapt to living in a highly active industrial environment. This paper presents a new approach to quantifying the anthropogenic impact on the marine mammal population. The results for sperm and beaked whales of a case study of regional population dynamics trends after the Deepwater Horizon oil spill, derived from passive acoustic-monitoring data gathered before and after the spill in the vicinity of the accident, are presented.


Murine antibody responses following systemic or mucosal immunization with viable or inactivated Vibrio cholerae.
Protocols are described for the induction of strong, consistent serum and mucosal antibody responses to Vibrio cholerae O1 or O139 lipopolysaccharide (LPS) following intranasal or oral immunization of adult mice with viable or formalin-killed bacteria. A simplified two-dose schedule for intranasal immunization has been identified, whereby viable bacteria elicit strong serum responses and, most importantly, also induce significant, sustained intestinal IgA responses. Using higher doses of bacteria it was also possible to generate consistently high intestinal and serum anti-LPS responses by the oral route. The efficacy of these immunization schedules was not dependent on co-administration of adjuvant. Gut responses were estimated using two sampling techniques involving the collection of fresh faecal pellets or the preparation of intestinal tissue extracts. The significant correlation between these estimates validates the more convenient approach of measuring intestinal responses using faecal pellet extracts, which allows repeated sampling from the same animals. V. cholerae O1 and O139 were similarly immunogenic by either mucosal route. More intensive immunization schedules for administration of formalin-killed bacteria have also been defined. Using these regimes it was possible to generate serum and gut antibody responses comparable to those elicited by viable V. cholerae. The established immunization protocols will allow evaluation of the systemic and mucosal immunogenicity of new vaccine formulations.


Disseminated Sporothrix schenckii infection with arthritis in a patient with acquired immunodeficiency syndrome.
Patients with the acquired immune deficiency syndrome are susceptible to a wide spectrum of opportunistic infections. We report a 34-year-old man who developed systemic sporotrichosis involving the skin and joints, and whose illness terminated in subacute encephalopathy and Pneumocystis carinii pneumonia. Fungal arthritis is another infection to which patients with this syndrome are subject.


Differentiation of viable and non-viable bacterial biofilms using electrorotation.
A new technique for studying the properties of biofilms has been developed, based on the phenomenon of electrorotation. Biofilms of Klebsiella rubiacearum were formed on the surfaces of 6 microns diameter polystyrene beads, and the presence of such films was found to alter their electrorotation spectra. The effects of adding a biocide (polyhexanide) to the surrounding aqueous medium was also investigated. The dielectric properties of the beads with biofilms, before and after biocide treatment, were interpreted from the electrorotation spectra using modelling methods that have been well tested for other heterogeneous biological systems. The technique is of value in understanding the physico-chemical properties of biofilms and can be adapted for monitoring the presence of toxic chemicals and for testing the activity of biocides against biofilms.


Optical imaging of cervical pre-cancers with structured illumination: an integrated approach.
Structured illumination microscopy is an inexpensive alternative to confocal microscopy that allows optical sectioning at a sub-cellular resolution. However, its application in imaging biological tissue has been limited by inadequate contrast present in them especially in reflectance imaging. Novel, optically active contrast agents like gold nanoparticles and quantum dots targeted against biomarkers of cancer can be integrated with structured illumination to image both the morphological and biochemical changes associated with epithelial pre-cancers. We modified the optical path of a widefield microscope to implement structured illumination both in reflectance and fluorescence modes. For imaging, we used 25-nm-diameter gold nanoparticles and CdSe quantum dots for reflectance and fluorescence imaging, respectively, to label three-dimensional tissue constructs of SiHa cervical cancer cells. Contrast agents were targeted against the epidermal growth factor receptor (EGFR) using an anti-EGFR monoclonal antibody. Agents targeted with a non-specific IgG antibody served as a control to monitor non-specific labeling. Our result shows that optically sectioned images taken with structured illumination are very comparable to those obtained using confocal microscopy. Moreover, images of three-dimensional cultures stained with the anti-EGFR agents show significantly more image intensity than those stained with the IgG targeted control. Our findings suggest that the combination of novel optical contrast agents and structured illumination can differentiate neoplastic cells which overexpress EGFR from normal cells in intact tissue. Combining structured illumination microscopy with novel contrast agents can potentially provide a powerful and inexpensive tool to aid in the detection of cervical pre-cancers.


Transmodulation of BCR signaling by transduction-incompetent antigen receptors: implications for impaired signaling in anergic B cells.
B cell tolerance can be maintained by functional inactivation, or anergy, wherein B cell Ag receptors (BCR) remain capable of binding Ag, but are unable to transduce signals. Although the molecular mechanisms underlying this unresponsiveness are unknown, some models of B cell anergy are characterized by disruption of proximal BCR signaling events, and by destabilization of the BCR complex. Receptor destabilization is manifest by a reduced ability to coimmunoprecipitate membrane Ig with the Ig-alpha/Ig-beta signal-transducing complex. To begin to explore the possibility that anergy is the consequence of receptor destabilization, we analyzed a panel of B lymphoma transfectants expressing constant amounts of signal-competent Ag receptors and varied amounts of a receptor with identical specificity, but bearing mutations that render it incapable of interacting with Ig-alpha/Ig-beta. This analysis revealed that coaggregation of signal-incompetent receptors prevented Ag-induced Ig-alpha and Syk phosphorylation, mobilization of Ca(2+), and the up-regulation of CD69 mediated by competent receptors. In contrast, Ag-induced Cbl and Erk phosphorylation were unaffected. Data indicate that coaggregation of destabilized receptors (as few as approximately 15% of total) with signal-competent receptors significantly affects the ability of competent receptors to transduce signals. Thus, BCR destabilization may underlie the Ag unresponsiveness of anergic B cells.


Assessment of the role of translationally controlled tumor protein 1 (TPT1/TCTP) in breast cancer susceptibility and ATM signaling.
The translationally controlled tumor protein 1 (TPT1/TCTP) has been implicated in the intracellular DNA damage response. We tested the role of TPT1 in breast cancer (BC) predisposition and re-evaluated its function in Ataxia-Telangiectasia mutated (ATM)-mediated damage recognition and DNA repair. The TPT1 coding sequence was scanned for mutations in genomic DNA from 200 breast cancer patients. TPT1 was down-regulated through siRNA in breast epithelial and fibroblast cell cultures. ATM activation after irradiation (IR) was analyzed by western blotting, and γH2A.X foci were monitored by immunocytochemistry. The sequencing study identified a novel, potentially damaging missense mutation in a single patient. Silencing of TPT1 did not significantly affect ATM kinase activity and did not impair the initial formation of γH2A.X foci, while we observed a marginally significant effect on residual γH2A.X foci at 6-48 h after IR. TPT1 does not harbor common mutations as BC susceptibility gene. Consistently, TPT1 protein is not required for the recognition of radiation-induced DNA damage via the ATM-dependent pathway and has only slight impact on timely repair. These results may be important when considering TPT1 as a DNA damage marker.


Comprehensive Electrocardiographic Analysis of Lead Exposed Workers: An Arrhythmic Risk Assessment Study.
To evaluate electrocardiographic parameters which are related with atrial and ventricular arrhythmias measured from 12-lead surface electrocardiogram (ECG) in workers occupationally exposed to lead. Sixty lead-exposed workers and 60 healthy controls were enrolled. Twelve-lead surface ECG was recorded and measurements of P wave durations (Pmax, Pmin) and P wave dispersion (PWD), QT durations and dispersion (QTd), corrected QT (QTc), Tp-e interval, and Tp-e/QT ratio were analyzed. The lead-exposed and control groups were similar with respect to baseline demographic, laboratory, and transthoracic echocardiographic indices. PWD (26.3 ± 9.7 vs 22.0 ± 9.0 ms, P = 0.014), Pmin (89.9 ± 13.8 vs 79.2 ± 10.1 ms, P < 0.001), and Pmax (116.2 ± 15.0 vs 101.2 ± 14.2 ms, P < 0.001), QT maximum (377.0 ± 27.6 vs 364.9 ± 28.5 ms, P = 0.02), QTd (38.4 ± 16.5 vs 30.5 ± 12.4 ms, P = 0.004), Tp-e interval (78.9 ± 16.5 vs 70.3 ± 14.5 ms, P = 0.003), and Tp-e/QT ratio (0.22 ± 0.04 vs 0.20 ± 0.04, P = 0.013) were significantly higher in lead-exposed workers. QT minimum and QTc values did not differ significantly. QT maximum, QTd, and Tp-e/QT ratio were correlated with urine lead level and Tp-e interval was correlated with both blood and urine lead levels. Lead-exposed workers have a higher risk for atrial and ventricular arrhythmias even without overt cardiac diseases compared with healthy subjects. These workers should be followed closely for adverse cardiovascular outcomes especially arrhythmias.


Correlation of narrow-band imaging findings using the Ni and European Laryngeal Society classification systems during in-office flexible laryngoscopy with histopathology.
This study used the European Laryngeal Society (2016) and Ni (2011 and 2019) classifications for narrow-band imaging and correlated the findings with histopathology. Retrospective analysis was conducted by retrieving data of patients who underwent micro-laryngoscopy for suspicious glottic lesions. The narrow-band imaging findings were classified using both classification systems. Retrieved histopathology report findings were correlated with narrow-band imaging data. Using the European Laryngeal Society and Ni classifications, 37 (69.8 per cent) and 35 (66 per cent) patients, respectively, were suspected to have malignant lesions. Upon histopathology, 37 (69.8 per cent) lesions were malignant. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy using the European Laryngeal Society classification were 91.9 per cent, 81.3 per cent, 91.9 per cent, 81.3 per cent and 88.7 per cent, and using the Ni classification were 91.9 per cent, 93.8 per cent, 97.1 per cent, 83.3 per cent and 92.5 per cent, respectively. The Ni classification had better specificity and accuracy. The European Laryngeal Society classification is simple to use and may serve as a useful screening tool. For optimum results, both European Laryngeal Society and Ni classifications may be used together, in that order.


In silico and in vitro studies to elucidate the role of 1HYN and 1QKI activity in BPA induced toxicity and its amelioration by Gallic acid.
Bisphenol A (BPA) is well known as an artificial environmental endocrine disrupting (ED) chemical. BPA also leads to many deleterious impacts on human blood through the production of reactive oxygen species and by some unknown mechanism. Up to now, very few studies have been conducted to assess the impact of BPA on Red Blood Corpuscle (RBC), Complete Blood Count (CBC), and no study on 1HYN (Erythrocyte Band 3 membrane protein) and 1QKI (Human Glucose 6 Phosphate Dehydrogenase) have been so far carried out. Besides, no study has been conducted to assess the ameliorating impact of the most commonly available antioxidant like Gallic Acid (GA). The present investigation revealed that BPA exposure (50-200 μg ml-1) causes significant increase in percent hemolysis and morphological alteration of RBC, as well as significant reduction in CBC except White Blood Cell (WBC), Platelet, and Red blood density width (RDW). BPA exposure also caused a significant reduction in G6PD activity. In silico docking study revealed that BPA effectively binds with 1HYN and 1QKI protein to alter its activity. Concurrent addition of GA (10-50 μg ml-1) with highest dose of BPA (200 μg ml-1) ameliorates all parameters significantly as compared to BPA (200 μg ml-1) treatment. Ameliorating effect of GA is mainly due to its antioxidant property and interaction with BPA, was confirmed using UV-VIS-NIR Spectrophotometric, molecular dynamic simulation and docking approach by YASARA software.


Anticipating change: how many acute care hospital beds will Manitoba regions need in 2020?
Being able to anticipate future needs for health services presents a challenge for health planners. Using existing population projections, two models are presented to estimate the demand for hospital beds in regions of Manitoba in 2020. The first, a current-use projection model, simply projects the average use for a recent 3-year period into the future. The second, a 10-year trend analysis, uses Poisson regression to project future demand. The current-use projection suggests a substantial increase in the demand for hospital beds, while the trend analysis projects a decline. The last projections are consistent with ongoing increases in rates of day surgeries and declines in lengths of stay. The current-use projections need to be considered in the context of relatively low occupancy rates in rural hospitals and previous research on appropriateness of stays in acute care hospitals. If measures are taken to ensure more appropriate use of acute care hospital beds in the future, then the current-use projections of bed shortages are not a cause for concern.


The Pain Divide: a cross-sectional analysis of chronic pain prevalence, pain intensity and opioid utilisation in England.
Our central research question was, in England, are geographical inequalities in opioid use driven by health need (pain)? To answer this question, our study examined: (1) if there are regional inequalities in rates of chronic pain prevalence, pain intensity and opioid utilisation in England; (2) if opioid use and chronic pain are associated after adjusting for individual-level and area-level confounders. Cross-sectional study design using data from the Health Survey for England 2011. England. Chronic pain prevalence, pain intensity and opioid utilisation. Participant data relating to chronic pain prevalence, pain intensity and opioid usage data were obtained at local authority level from the Health Survey for England 2011; in total, 5711 respondents were included in our analysis. Regional and local authority data were mapped, and a generalised linear model was then used to explore the relationships between the data. The model was adjusted to account for area-level and individual-level variables. There were geographical variations in chronic pain prevalence, pain intensity and opioid utilisation across the English regions-with evidence of a 'pain divide' between the North and the South, whereby people in the North of England more likely to have 'severely limiting' or 'moderately limiting' chronic pain. The intensity of chronic pain was significantly and positively associated with the use of opioid analgesics. There are geographical differences in chronic pain prevalence, pain intensity and opioid utilisation across England-with evidence of a 'pain divide'. Given the public health concerns associated with the long-term use of opioid analgesics-and their questionable activity in the management of chronic pain-more guidance is needed to support prescribers in the management of chronic pain, so the initiation of opioids can be avoided.


Isolation and characterization of 36 polymorphic microsatellite markers in Schima superba (Theaceae).
Our objective was to develop microsatellite markers to investigate the level of genetic diversity within and among populations in a dominant evergreen broad-leaved tree, Schima superba, in southern China. Thirty-six microsatellite markers were developed and showed polymorphism in three populations. The number of alleles per locus ranged from six to 34, with an average of 19. The observed and expected heterozygosities ranged from 0.242 to 1.000 and from 0.504 to 0.945, respectively. The developed microsatellites will be useful for studying genetic diversity and population structure in S. superba.


Effects of long-term nasal continuous positive airway pressure on C-reactive protein in patients with obstructive sleep apnea syndrome.
Serum levels of C-reactive protein (CRP) were measured in 96 consecutive patients with obstructive sleep apnea syndrome (OSAS) before and after nasal continous positive airway pressure treatment. CRP levels only displayed significant correlations with body mass index (BMI) before treatment. No significant changes were observed in BMI and CRP levels after 9 months of treatment. These data suggest that CRP levels in patients with OSAS may be associated with obesity rather than OSAS itself.


Unexpected absence of a myeloid surface antigen (3-fucosyl-N-acetyllactosamine) in promyelocytic leukemia.
According to the criteria of the FAB classification we diagnosed acute promyelocytic leukemia in 19 out of 191 AML blast cell populations. The reaction pattern with the anti-myeloid monoclonal antibodies VIM2, MCS2 and MY9 and the MHC-class II antibody VID1 in all cases showed the characteristics which were expected from the normal granulopoiesis. The monoclonal antibody VIM-D5, which recognizes the myeloid cell surface structure 3-fucosyl-N-acetyllactosamine (3-FAL), was not reactive with the majority of promyelocytic blast cells, however. In 13 cases 3-FAL was not detectable and in 6 cases only low or moderate reactivity of VIM-D5 with blast cells was observed. The phenotype of leukemic promyelocytes thus differs from the majority of normal promyelocytes in this respect.


Factors that Affect the Adherence to ADHD Medications during a Treatment Continuation Period in Children and Adolescents: A Nationwide Retrospective Cohort Study Using Korean Health Insurance Data from 2007 to 2011.
Several factors, such as male gender, older age, type of insurance, comorbid conditions, and medication type, have been associated with attention-deficit/hyperactivity disorder (ADHD) medication adherence rates, but the results have been inconsistent. We analyzed data to answer several questions: 1) How old were patients who first refilled their treatment medications used primarily for ADHD, regardless of the medication type? 2) What socio-demographic factors are associated with medication adherence? 3) What medical conditions, such as medication type and comorbid diagnosis, influence adherence? We analyzed National Health Insurance data, which comprised continuously enrolled Korean National Medical Insurance children (6-18 years) with at least 2 ADHD prescription claims (January 2008-December 2011). The persistence of use regarding the days of continuous therapy without a 30-day gap were measured continuously and dichotomously. Adherence, using a medication possession ratio (MPR), was measured dichotomously (80% cut-off). The cumulative incidence of index cases that initiated medication refills for ADHD treatment during the 4 year period was 0.85%. The patients who exhibited a MPR greater than 80 comprised approximately 66%. The medication type, high school age groups, physician speciality, treatment at a private clinic, and comorbid conditions were associated with medication adherence during continuous treatment using a multivariate analysis. A better understanding of ADHD treatment patterns may lead to initiatives targeted at the improvement of treatment adherence and persistence. Other factors, including the severity, family history, costs, type of comorbidities, and switching patterns, will be analyzed in future studies.


Effects of Estradiol (-17beta) on learning, memory and cerebral energy metabolism in male rats after intracerebroventricular administration of streptozotocin.
Treatment of adult rats with intracerebroventricularly (i.c.v.) injected streptozotocin (STZ) may provide a relevant animal model of chronic neuronal dysfunction that is characterized by a decrease in both the neuronal metabolism of glucose and the formation of energy. The present study was designed to evaluate whether or not rats treated with triplicate i.c.v. STZ injection would show long-term effects in learning and memory behavior as measured by means of a holeboard test, closed field activity, and passive avoidance behavior over a period of 6 weeks. For this purpose, animals with good performance were discriminated from those with poor performance by the holeboard test before i.c.v. administration of STZ. After a 1-week training period with the holeboard all animals improved their abilities in learning and memory, and the improvement was maintained over the investigation period of 6 weeks in the control group. After i.c.v. STZ working memory (WM), reference memory (RM), as well passive avoidance (PA) behavior decreased, deteriorating progressively during the investigation period. The latter were accompanied by a permanent deficit in cerebral energy metabolism. The ongoing deterioration in behavior and the sustained deficit in cerebral energy metabolism occurring after a triplicate i.c.v. STZ administration lead us to assume that this animal model may be appropriate for the investigation of mechanisms characteristic for sporadic Alzheimer disease. In this context, the effect of Estradiol-17beta (E2) on behavior and energy metabolism was studied. We found that E2 slowed down the i.c.v. STZ-induced deterioration in memory functions, partially compensated the learning deficit, and improved the disturbances in cerebral energy metabolism to the extent that it was almost completely normal again. These findings underscore the beneficial effect of E2 in dementia disorders.


The use of biomarkers and HPV genotyping to improve diagnostic accuracy in women with a transformation zone type 3.
Twenty percent of women referred to colposcopy have a type 3 transformation zone-where colposcopic assessment for high-grade dysplasia (CIN2+) is not possible. This study examines the effectiveness of HPV biomarkers and genotyping in combination with techniques that sample an endocervical TZ. A prospective diagnostic accuracy study. Women booked for large-loop excision (LLETZ) with squamous dyskaryosis, high-risk HPV and a TZ3 were recruited. Immediately prior to LLETZ samples were collected for p16/Ki-67 dual-stained cytology, HPV genotyping and H&E, p16- and Ki-67-stained endocervical curettings. In women with low-grade screening (n = 64), 35.9% had CIN2+; dual-stained cytology had the greatest effect on the PPV of routine screening (76.1% vs 35.9%) and perfectly predicted the absence of CIN2+. In women with a high-grade screening result (n = 37); 75.6% had CIN2+ and dual-stained curettings improved the PPV (96.5 vs 75.6%). With high-grade screening and a TZ3, LLETZ appears safest as three quarters have CIN2+ . Women with low-grade screening and a TZ3 have a twofold increased risk of CIN2+ when compared to women where the TZ is visible. The use of dual-stained cytology may help identify those women who can be safely offered surveillance and those who require treatment.


Ongoing challenges in access to diabetes care among the indigenous population: perspectives of individuals living in rural Guatemala.
Indigenous persons living in Latin America suffer from a higher prevalence of type 2 diabetes compared to their non-indigenous counterparts. This difference has been attributed to a wide range of factors. Future interventions could be influenced by a deeper understanding of the challenges that impact care in rural regions and in other low-income settings. This study was conducted using a modified grounded theory approach. Extended observations and fifteen interviews were performed with adult male and female residents of three rural Mayan towns in Sololá Department, Guatemala using purposive sampling. Questions focused on the perceptions of individuals living with type 2 diabetes and their caregivers regarding disease and treatment. Across interviews the most common themes that emerged included mistreatment by healthcare providers, mental health comorbidity, and medication affordability. These perceptions were in part influenced by indigeneity, poverty, and/or gender. Both structural and cultural barriers continue to impact diabetes care for indigenous communities in rural Guatemala. The interviews in this study suggest that indigenous people experience mistrust in the health care system, unreliable access to care, and mental health comorbidity in the context of type 2 diabetes care. These experiences are shaped by the complex relationship among poverty, gender, and indigeneity in this region. Targeted interventions that are conscious of these factors may increase their chances of success when attempting to address similar health disparities in comparable populations.


Ethnoecology - The best medicine against allergy?
This essay, which is the 6(th) in the series "Recollections, Reflections, and Revelations: Ethnobiologists and Their First Time in the Field", is a personal reflection by the researcher on his first field experiences with ethnobiology. Author writes on how Hungarian herders in the Hortobágy salt steppes and Csángó people in the Carpathian mountains changed his views on landscape, vegetation, local people, traditional small-scale grassland management and finally, ecology and nature conservation.


Targeting dendritic cells with antigen via dendritic cell-associated promoters.
The induction of tumor-specific immune responses is largely dependent on the ability of dendritic cells (DCs) to present tumor-associated antigens to T lymphocytes. Therefore, we investigated the use of DC-associated promoter-driven genetic vaccines to specifically target DC in vivo. Restricted expression of vaccine-encoding genes in DC should enhance specificity and improves their safety for clinical applications. Hereto, 3-5 kb upstream sequences of the murine genes encoding CD11c, DC-SIGN, DC-STAMP and Langerin were isolated, characterized and subcloned into enhanced green fluorescent protein (EGFP) reporter constructs. Upon electroporation, EGFP was expressed in DC cell lines, but not in other cell lines, confirming DC-restricted promoter activity. When these promoters were cloned into a construct upstream of the gene for ovalbumin (OVA), it appeared that DC-STAMP promoter-driven expression of OVA (pDCSTAMP/OVA) in DC yielded the most efficient OVA-specific CD4+ and CD8+ T-cell responses in vitro. Administration of pDC-STAMP/OVA in vivo, using the tattoo gun vaccination system, evoked specific immune responses as evidenced in a mouse tumor model. Adoptively transferred pDC-STAMP/OVA-transfected DCs induced strong CD8+ T-cell proliferation in vivo. These experiments demonstrate that our DC-directed promoter constructs are potential tools to restrict antigen expression in DC and could be implemented to modulate DC function by the introduction of relevant proteins.


Study on degradation kinetics of 2-(2-hydroxypropanamido) benzoic acid in aqueous solutions and identification of its major degradation product by UHPLC/TOF-MS/MS.
A RP-HPLC method was developed and validated for the degradation kinetic study of 2-(2-hydroxypropanamido) benzoic acid (HPABA), a promising anti-inflammatory drug, which would provide a basis for further studies on HPABA. The effects of pH, temperature, buffer concentration and ionic strength on the degradation kinetics of HPABA were discussed. Experimental parameters such as degradation rate constants (k), activation energy (Ea), acid and alkali catalytic constants (k(ac), k(al)), shelf life (t1/2) and temperature coefficient (Q10) were calculated. The results indicated that degradation kinetics of HPABA followed zero-order reaction kinetics; degradation rate constants (k) of HPABA at different pH values demonstrated that HPABA was more stable in neutral and near-neutral conditions; the function of temperature on k obeyed the Arrhenius equation (r = 0.9933) and HPABA was more stable at lower temperature; with the increase of ionic strength and buffer concentration, the stability of HPABA was decreased. The major unknown degradation product of HPABA was identified by UHPLC/TOF-MS/MS with positive electrospray ionization. Results demonstrated that the hydrolysis product was the primary degradation product of HPABA and it was deduced as anthranilic acid.


Biallelic KCTD3 nonsense variant derived from paternal uniparental isodisomy of chromosome 1 in a patient with developmental epileptic encephalopathy and distinctive features.
A biallelic nonsense variant of the potassium channel tetramerization domain-containing protein 3 gene (KCTD3) [c.1192C>T; p.R398*] was identified in a patient with developmental epileptic encephalopathy with distinctive features and brain structural abnormalities. The patient showed isodisomy of chromosome 1, where KCTD3 is located, and the father was heterozygous for the same variant. Based on these findings, paternal uniparental disomy was considered to cause the biallelic involvement of KCTD3.


Hydronephrosis: unrecognized complication following restorative proctocolectomy with ileal pouch-anal anastomosis.
Restorative proctocolectomy with ileal pouch-anal anastamosis (IPAA) is the operation of choice to avoid permanent ileostomy in the surgical management of patients with ulcerative colitis and familial adenomatous polyposis coli. IPAA with two loops of small intestine (J-pouch) is the most common configuration. Mortality rates are low, and despite significant morbidity patient satisfaction remains high. Functional results are good in most cases. We present two cases of hydronephrosis following IPAA. Loin pain was the presenting symptom in both cases, with one patient developing impaired renal function. Hydronephrosis has not been reported following IPAA, but with other forms of pelvic and retroperitoneal surgery it is believed to remain largely asymptomatic. We discuss the etiology of this previously unrecognized complication and suggest that it may be more prevalent following IPAA than is realized.


Chromatin structural transitions following histone H1 displacement by phosphatidylserine vesicles and low pH treatment. A multiparametric analysis involving flow cytometry, electron microscopy, and nuclease digestion.
We describe several morphological and functional modifications in isolated rat liver nuclei incubated in the presence of phosphatidylserine (PS) multilamellar vesicles (MLV). These effects, which occur through the release of histone H1, induce chromatin decondensation, as shown by electron microscopy and nuclease digestion. Flow cytometry was employed to monitor these changes in chromatin structure in isolated nuclei by means of perpendicular light scatter (PLS) and fluorescence signals. Chromatin decondensation induced by PS or by low pH treatment was accompanied by an increase in perpendicular light scatter and by less efficient binding of ethidium bromide. These flow cytometric findings are peculiar to chromatin decondensation induced by displacement of histone H1. Conversely, chromatin decondensation caused by lowering of the divalent ion concentration, without displacement of histone H1, is characterized only by an increase in perpendicular light scatter.


Effect of zearalenone on some physiological processes of maize seeds (Zea mays L var Suwan composite).
Physiological processes of maize seeds (Zea mays L var Suwan composite) were found to be affected by five different concentrations (linear relationship between 100 and 2,000 μ/L) of zearalenone and duration of soaking le 20 h. Inhibitions in seed germination (14-58 %), shoot length (16-61 %), root length (9-76 %), total chlorophyll (6-45 %), and carotenoid (5-73 %) were evident due to above treatments.


[Tinnitus].
Tinnitus is an auditory perception without adequate external acoustic sources. The incidence of tinnitus in the general population is high, and can lead to total decompensation in some of the affected patients. Tinnitus is a symptom of an unspecific lesion of the ear or the central auditory system. In many cases, the cause of tinnitus can be inferred from the medical history and from the results of specific medical examines. Nevertheless, in many cases there is still no causal therapy available. As a result, for most patients approaches such as proper counselling, the fitting of hearing aids or tinnitus retraining therapy are adopted.


The use of isochronal reference standards in phase and modulation fluorescence lifetime measurements.
Errors in phase and modulation lifetime measurements observed with the only commercially available instrument are readily apparent when the Debye-Sears modulation tank is not perfectly tuned. Unfortunately, we have found that exact tuning was extremely difficult to achieve and maintain. We report that these errors could be reduced by using single-lifetime (homogeneous) reference standards whose fluorescence lifetime approximated that of the unknown sample (isochronal standards). A number of useful standards are suggested. In the proposed method, the phase shift and relative modulation of the sample emission are measured using the isochronal standard as a reference to determine the effective characteristics of the sinusoidal excitation. The importance of the improvement in accuracy accomplished by the proposed methods is illustrated by the accurate resolution of fluorescence lifetime heterogeneity for two known heterogeneous samples.


The influence of bone marrow- and synovium-derived mesenchymal stromal cells from osteoarthritis patients on regulatory T cells in co-culture.
There is increasing evidence that inflammation in the synovium plays a major role in the progression of osteoarthritis (OA). However, the immunogenic properties of mesenchymal stromal cells (MSCs), which are considered to regulate immunity in various diseases, remain largely unknown in OA. The purpose of this study was to determine the influence of MSCs from OA patients on regulatory T cells (Tregs ) in an allogeneic co-culture model. Bone marrow (BM) and synovial membrane (SM) were harvested from hip joints of OA patients and co-cultured with lymphocytes enriched in CD4(+) CD25(+) CD127(-) regulatory T cells (Treg (+) LC) from healthy donors. Treg proportions and MSC markers were assessed by flow cytometry. Cytokine levels were assessed after 2 and 5 days of co-cultivation. Additionally, Treg (+) LC cultures were analysed in the presence of interleukin (IL)-6 and MSC-supernatant complemented medium. B-MSCs and S-MSCs were able to retain the Treg proportion compared to lymphocyte monocultures. T cell-MSC co-cultures showed a significant increase of IL-6 compared to MSC cultures. S-MSCs produced higher amounts of IL-6 compared to B-MSCs, both in single and T cell co-cultures. The effect of retaining the Treg percentage could be reproduced partially by IL-6 addition to the medium, but could only be observed fully when using MSC culture supernatants. Our data demonstrate that retaining the Treg phenotype in MSC-T cell co-cultures can be mediated by MSC derived from OA patients. IL-6 plays an important role in mediating these processes. To our knowledge, this study is the first describing the interaction of MSCs from OA patients and Tregs in an allogeneic co-culture model.


Differential expression of cell survival and cell cycle regulatory proteins in cutaneous squamoproliferative lesions.
Previous models of cutaneous carcinogenesis have primarily focused on the regulation of keratinocyte (KC) proliferation and differentiation. However, it has become clear in many neoplastic systems that altered rates of cell death and/or inability to undergo growth arrest can also contribute to the development of cancer. Apoptosis-regulatory proteins include those that block apoptosis such as Bcl-2 and Bcl-x, whilst a related protein Bax promotes apoptosis. Cell cycle regulatory proteins include those associated with growth arrest, i.e. p21wafl, p53, and those associated with proliferation, i.e. Ki-67. Paraffin embedded samples from ten different lesions of squamous cell carcinoma (SCC), Bowen's disease (BD), keratoacanthomas (KA), and nine normal adult skin samples were stained by immunohistochemistry to detect expression of Bcl-2, Bcl-x, Bax, Ki-67, p21wafl, p53 and apoptosis (TUNEL assay). Compared to low levels of Bcl-x and Bcl-2 immunostaining in normal skin, all the squamoproliferative lesions had strong and diffuse KC expression of Bcl-x (>80%) but minimal to absent KC Bcl-2 expression (<15%). Bax immunopositivity was limited to the basal layer in normal skin and BD. In contrast, by examining serial sections both Bcl-x and Bax appeared to be coexpressed by the majority of malignant KCs in KA and SCC (>70%). These immunostaining profiles reveal that squamoproliferative lesions, including invasive transformed KCs, preferentially express Bcl-x over Bcl-2, in addition to upregulating their Bax levels. Even though there were numerous TUNEL positive cells in these squamoproliferative lesions, no other evidence of apoptosis was seen reinforcing the necessity to use caution when relying on TUNEL staining for identification of programmed cell death in skin biopsies. Normal sun-exposed skin had low but detectable p53 and rare p21wafl KC expression. Significantly higher numbers of p21wafl and p53 immunopositive KCs were noted throughout the lesions in BD and SCC in contrast to KA where p53 and rare p21wafl immunopositive KCs were primarily limited to the periphery of the tumor cell islands. In general, p53 KC expression was higher in all squamoproliferative lesions and sun-exposed normal skin compared to p21Wafl expression. Summary of the expression of cell cycle regulatory proteins for both p21wafl and p53 KC expression was: SCC > BD > KA, in marked contrast to Ki-67 KC expression which was: BD > KA > SCC. The relatively few malignant cells in SCC that were actively participating in the cell cycle (i.e. Ki-67 positive) suggests that these neoplasms may arise primarily by increased cell survival and resistance to apoptosis rather than by hyperproliferation. These studies emphasize the importance of examining multiple members of protein families that regulate apoptosis, proliferation, growth arrest, and differentiation. It is the overall balance between these cellular phenomena that determine whether a cell remains viable or undergoes programmed cell death and contributes to the appearance of a neoplasm. The overexpression of Bcl-x may confer a survival advantage to malignant KCs unable to growth arrest to repair damaged DNA (mutant p53) and/or undergo terminal differentiation (increased p21wafl). Thus, mutation or aberrant expression of such proteins may participate in the multistep process of carcinogenesis that gives rise to these squamoproliferative lesions.


[Treatment of diaphyseal pseudoarthrosis with transosseous osteosynthesis by the Ilizarov method. Radiographic signs of reparative osteogenesis].
The main phases of ossification are shown-based on the results achieved by re-examining the X-ray pictures of 14 patients affected by tubular bones pseudoarthrosis treated with Ilizarov's closed compression-distraction osteosynthesis. The roentgen signs in particular are examined which are considered as expressive of positive evolution of the pseudoarthrosis. An outline of X-ray evolution of nonunions is also proposed, referring to the peculiar biological characters of ossification under compression-distraction stimulation.


Associations of insomnia symptoms with subsequent health services use among community-dwelling U.S. older adults.
Determine the association of insomnia symptoms with subsequent health services use, in a representative sample of U.S. older adults. Participants were 4,289 community-dwelling Medicare beneficiaries who had continuous fee-for-service Medicare coverage 30 days before, and 1 year after the National Health and Aging Trends Study (NHATS) Round 1 interview. Participants reported past-month insomnia symptoms (i.e. sleep onset latency >30 min, difficulty returning to sleep) which we categorized as 0, 1, or 2 symptoms. Outcomes were health services use within 1 year of interviews from linked Medicare claims: emergency department (ED) visits, hospitalizations, 30-day readmissions, home health care (all measured as yes/no), and number of hospitalizations and ED visits. Overall, 18.5% of participants were hospitalized, 28.7% visited the ED, 2.5% had a 30-day readmission, and 11.3% used home health care. After adjustment for demographics, depressive and anxiety symptoms, medical comorbidities, and BMI, compared to participants with no insomnia symptoms, those with two insomnia symptoms had a higher odds of ED visits (odds ratio [OR) = 1.60, 95% confidence interval [CI] = 1.24-2.07, p < 0.001), hospitalizations (OR = 1.29, 95% CI = 1.01-1.65, p < 0.05), and 30-day readmissions (OR = 1.88, 95% CI = 1.88-3.29, p < 0.05). Reporting 2 insomnia symptoms, versus no insomnia symptoms, was associated with a greater number of ED visits and hospitalizations (incidence rate ratio (IRR) = 1.52, 95% CI = 1.23-1.87, p < 0.001; IRR = 1.21, 95% CI = 1.02-1.44, p < 0.05, respectively) after adjusting for demographic and health characteristics. Among older adults, insomnia symptoms are associated with greater health services use, including emergency department use, hospitalization, and 30-day readmission. Targeting insomnia may lower health services use.


Estimated Resting Metabolic Rate and Body Composition Measures Are Strongly Associated With Diabetic Retinopathy in Indonesian Adults With Type 2 Diabetes.
To investigate the associations of estimated resting metabolic rate (RMR), body fat (BF), subcutaneous fat (SCF), visceral fat (VF), fat-free mass (FFM) percentage, BMI, and waist circumference (WC) with diabetic retinopathy (DR) in Indonesian adults with type 2 diabetes. This was a community-based cross-sectional study of 1,184 subjects with type 2 diabetes. DR was assessed from fundus photography and categorized as mild, moderate nonproliferative DR (NPDR), and vision-threatening DR (VTDR). RMR and body composition parameters were measured using automated body composition scan. Logistic regression with semipartial correlation analysis was used. DR and VTDR were present in 43.1 and 26.3% of participants, respectively. After adjustment for age, sex, diabetes duration, fasting glucose, systolic blood pressure, smoking, diabetic ulcer, and use of combined diabetes treatment, per SD increase in RMR (odds ratio [OR] 2.60 [95% CI 2.19-3.07]; P < 0.001) was associated with DR, while per SD increases in BF (0.66 [95% CI 0.56-0.78]; P < 0.001), FFM (0.69 [0.57-0.84]; P < 0.001), VF (0.77 [0.67-0.88]; P < 0.001), BMI (0.83 [0.73-0.94]; P = 0.004), and WC (0.81 [0.73-0.91]; P < 0.001) were inversely associated with presence of DR. Similar associations were found for VTDR. Among all variables, RMR had the largest contribution to the variance in the DR model (39%). In this study, RMR and body composition measures were strongly associated with and contributed considerably to the presence and severity of DR. These findings, if confirmed, suggest that RMR and body composition may be strong markers that represent actual metabolic state in the pathophysiology of DR.


A unique reactive residue in adenosine triphosphate phosphoribosyltransferase sensitive to five conformation and dissociation states.
Adenosine triphosphate phosphoribosyltransferase is inactivated rapidly by bulky alkylating reagents in a biphasic reaction. The initial inactivation rate is dependent upon an optimal concentration of histidine and is more rapid at low enzyme concentrations and low ionic strength. A histidine-free dimer form of the enzyme is the proposed reactive species. The dimer is shown by ultraviolet difference spectroscopy to bind histidine about 1 order of magnitude more weakly than the hexameric form of the enzyme. Alkylated enzyme is similar to native enzyme in dissociation and histidine-binding properties. Native enzyme must exist at significant levels in at least five different conformational and dissociative states to account for the inactivation behavior.