The impact of ultra-high-density mapping on long-term outcome after catheter ablation of ventricular tachycardia.
Ultra-high-density (UHD) mapping can improve scar area detection and fast activation mapping in patients undergoing catheter ablation of ventricular tachycardia (VT). The aim of the present study was to compare the outcome after VT ablation guided by UHD and conventional point-by-point 3D-mapping. The acute and long-term ablation outcome of 61 consecutive patients with UHD mapping (64-electrode mini-basket catheter) was compared to 61 consecutive patients with conventional point-by-point 3D-mapping using a 3.5 mm tip catheter. Patients, whose ablation was guided by UHD mapping had an improved 24-months outcome in comparison to patients with conventional mapping (cumulative incidence estimate of the combination of recurrence or disease-related death of 52.4% (95% confidence interval (CI) [36.9-65.7]; recurrence: n = 25; disease-related death: n = 4) versus 69.6% (95% CI [55.9-79.8]); recurrence: n = 31; disease-related death n = 11). In a cause-specific Cox proportional hazards model, UHD mapping (hazard ratio (HR) 0.623; 95% CI [0.390-0.995]; P = 0.048) and left ventricular ejection fraction > 30% (HR 0.485; 95% CI [0.290-0.813]; P = 0.006) were independently associated with lower rates of recurrence or disease-related death. Other procedural parameters were similar in both groups. In conclusion, UHD mapping during VT ablation was associated with fewer VT recurrences or disease-related deaths during long-term follow-up in comparison to conventional point-by-point mapping. Complication rates and other procedural parameters were similar in both groups.


Combining continuous flow aerobic granulation using an external selector and carbon-efficient nutrient removal with AvN control in a full-scale simultaneous nitrification-denitrification process.
The James R. Dolorio Water Reclamation Facility in Pueblo, Colorado, uses AvN aeration controls to lower aeration energy while promoting carbon-efficient nutrient removal and hydrocyclone-based wasting to achieve SVI improvements and process intensification. The results from the full-scale installation showed that hydrocyclone-based wasting helped improve settling characteristics by reducing the SVI from 200 ± 52 mL/g to 83 ± 22 mL/g within weeks of operation. PAO and nitrifiers were preferentially retained in dense flocs and granules, while lighter heterotrophic and filamentous organisms were preferentially wasted, thus uncoupling the SRT of these two fractions relative to the overall SRT. The SRT was estimated at 14.4 ± 3.4 days for dense aggregates and 7.1 ± 2.3 days for lighter flocs. The use of AvN control with continuous low DO conditions resulted in low DO conditions (< 0.3 mgO2/L) reducing air demand by 50% while providing excellent nitrogen (effluent TIN < 11 mgN/L) and TP removal (effluent TP < 1 mgP/L) at low primary effluent COD/N ratio of 6.0. The presence of comammox was demonstrated through molecular analysis, while ex-situ batch tests revealed the presence of DPAO, which could have attributed to the energy and carbon-efficient biological nutrient removal.


Autoimmune manifestations in common variable immunodeficiency.
About 20% of subjects with common variable immune deficiency (CVID) develop an autoimmune complication, most often immune thrombocytopenia or hemolytic anemia. While the pathogenesis of autoreactivity is unknown for CVID subjects in general, and to a greater extent in those with autoimmunity, there is a loss of switched memory B cells. About 7-8% of CVID subjects have mutations in the transmembrane activator and calcium-modulating cyclophilin ligand interactor (TACI), a significant association with this immune defect, although the same mutations may be found in normal relatives and rarely in healthy blood donors. In addition to generalized B cell dysfunction, defective elimination of autoimmune B cells has been demonstrated.


Anterior capsule dyes and labeled viscoelastic solutions to enhance contrast in EAS-1000 Scheimpflug images.
To enhance contrast in images produced by the EAS-1000 to facilitate evaluation of the rheological properties of viscoelastic substances during simulated cataract surgery. Department of Preclinical Ophthalmology, Scientific Affairs, Pharmacia AB, Uppsala, Sweden. Cataract surgery was performed in 200 fresh porcine cadaver eyes and documented by EAS-1000 Scheimpflug photography. The light scattering caused by anterior capsule dyes, labeled irrigation solutions, and labeled viscoelastic solutions was statistically analyzed in the images. Visualization by light scattering was significantly enhanced by a fluorescein labeled viscoelastic solution and by an eosin-Y- or phloxine-B-dyed capsule (P < .05). A side effect of eosin Y and phloxine B was that the capsule became fragile, making it difficult to perform a continuous curvilinear capsulorhexis. Fluorescein-labeled viscoelastic solutions are recommended to enhance contrast in the images when retention capacity is evaluated. Light scattering from capsule dyeing facilitated anterior chamber depth measurements in the images.


Managing Osteoarthritis and Other Chronic Musculoskeletal Pain Disorders.
Osteoarthritis (OA) is a common problem in society and can lead to significant disability and impairment of a patient's capacity to perform activities of daily living. The focus of this article is various treatment options for the management of OA, with emphasis on conservative management. The emphasis is on the role of exercise, pharmacology, intra-articular joint injections, and bracing options in the management of OA.


Nucleosidediphosphate kinase of Escherichia coli, a periplasmic enzyme.
The ATP-ADP exchange activity previously described in a membrane farction of Escherichia coli appeared after a cold osmotic shock according to Neu and Heppel ((1965) J. Biol. Chem. 240, 3685--3692) in the shock fluid. Membranes derived from shocked cells had no activity. The enzyme responsible for this activity has been purified 125-fold and catalyzed the transfer of a phosphoryl radical from ribonucleosidetriphosphates (NTPs) to ribonucleosidediphosphates (NDPs); this is, therefore, a non-specific nucleosidediphosphate kinase (ATP:nucleosidediphosphate phosphotransferase, EC 188.8.131.52). The activity required the presence of a divalent cation, Mg2+, Mn2+ or Ca2+ at a unity mol/mol ratio of nucleotide for maximal activation. The enzyme exhibited simple saturation kinetics with respect to the phosphate donor but inhibition by excess substrate was observed upon increasing phosphate acceptor. The kinetics of the reaction indicated an ordered bi-molecular ping-pong reaction mechanism. Differential heat sensitivity of the enzyme whether it is heated alone with ATP, ADP or Mg2+ opens possibilities to study different enzyme-substrate complexes.


Interacting pest control and pollination services in coffee systems.
Biodiversity-mediated ecosystem services (ES) support human well-being, but their values are typically estimated individually. Although ES are part of complex socioecological systems, we know surprisingly little about how multiple ES interact ecologically and economically. Interactions could be positive (synergy), negative (trade-offs), or absent (additive effects), with strong implications for management and valuation. Here, we evaluate the interactions of two ES, pollination and pest control, via a factorial field experiment in 30 Costa Rican coffee farms. We found synergistic interactions between these two critical ES to crop production. The combined positive effects of birds and bees on fruit set, fruit weight, and fruit weight uniformity were greater than their individual effects. This represents experimental evidence at realistic farm scales of positive interactions among ES in agricultural systems. These synergies suggest that assessments of individual ES may underestimate the benefits biodiversity provides to agriculture and human well-being. Using our experimental results, we demonstrate that bird pest control and bee pollination services translate directly into monetary benefits to coffee farmers. Excluding both birds and bees resulted in an average yield reduction of 24.7% (equivalent to losing US$1,066.00/ha). These findings highlight that habitat enhancements to support native biodiversity can have multiple benefits for coffee, a valuable crop that supports rural livelihoods worldwide. Accounting for potential interactions among ES is essential to quantifying their combined ecological and economic value.


Aureobasidium melanigenum catheter-related bloodstream infection: a case report.
Aureobasidium melanigenum is a ubiquitous dematiaceous fungus that rarely causes invasive human infections. Here, we present a case of Aureobasidium melanigenum bloodstream infection in a 20-year-old man with long-term catheter use. A 20-year-old man receiving home care with severe disabilities due to cerebral palsy and short bowel syndrome, resulting in long-term central venous catheter use, was referred to our hospital with a fever. After the detection of yeast-like cells in blood cultures on day 3, antifungal therapy was initiated. Two identification tests performed at a clinical microbiological laboratory showed different identification results: Aureobasidium pullulans from matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and Cryptococcus albidus from a VITEK2 system. Therefore, we changed the antifungal drug to liposomal amphotericin B. The fungus was identified as A. melanigenum by DNA sequence-based analysis. The patient recovered with antifungal therapy and long-term catheter removal. It is difficult to correctly identify A. melanigenum by routine microbiological testing. Clinicians must pay attention to the process of identification of yeast-like cells and retain A. melanigenum in cases of refractory fungal infection.


Glycosylation of the thrombin-like serine protease ancrod from Agkistrodon rhodostoma venom. Oligosaccharide substitution pattern at each N-glycosylation site.
In a previous study, we determined the structures of the glycans present in ancrod, a thrombin-like serine protease from the venom of the Malayan pit viper Agkistrodon rhodostoma (Pfeiffer et al. (1992) Eur J Biochem 205:961-78). In order to allocate the various carbohydrate chains to distinct N-glycosylation sites of the molecule, we have now isolated individual glycopeptides. Peptide moieties were identified after deglycosylation with peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F by amino acid analysis and sequencing. Liberated oligosaccharides were assigned to the previously deduced carbohydrate structures by high performance liquid chromatography. Although only quantitative differences were observed, the results indicate that each glycosylation site of ancrod carries its characteristic oligosaccharide pattern. Furthermore, all potential sites were shown to be substituted by carbohydrates.


[Giant aneurysm of the middle cerebral artery. Report of a case].
A case of giant aneurysm of the middle artery with the size of 12,0 x 8,0 x 6,0 cm is reported. Some aspects of the treatment of such aneurysm are considered. As far as we know this is the largest intracranial aneurysm ever recorded.


Protective Effects of N-acetylcysteine Niosome Nanoparticles on Paraquatinduced Nephrotoxicity in Male Rats.
Paraquat (PQ), as a bipyridyl compound, is widely used as an effective herbicide that produces reactive oxygen species (ROS), affecting the unsaturated lipids of cell membranes leading to cell mortality. N-acetylcysteine (NAC) is a medication that has a beneficial role in reducing the intoxication of kidneys caused by PQ. Niosomes are bilayer vesicles that enhance the bioavailability of drugs. This study aimed to compare the effects of NAC and niosome of NAC (NACNPs) on PQ-induced kidney toxicity concerning its antioxidant activity. In this experimental study, after formulating NACNP, 30 Wistar male rats weighing 180 to 250 gm were classified into five groups: the control group was treated with normal saline, while the other four groups received 35mg/kg/day of PQ via intraperitoneal route and, was treated with 25mg/kg/day NAC, 25mg/kg/day niosome and 25 mg/kg/day NACNP by gavage, Then, oxidative stress biomarkers such as total antioxidant capacity (TAC), catalase activity (CAT), lipid peroxidation (LPO), and total thiol group (TTG), plus blood urea nitrogen (BUN) and creatinine levels were evaluated in kidney tissue homogenate and examined histopathologically. The results revealed that TTG increased significantly in NAC & NACNP groups than in the PQ group. Further, in the PQ group, LPO increased significantly compared with the control, NAC, and NACNP groups, while in the NAC and NACNP group, LPO diminished compared with the PQ group. There was no significant difference in TAC between groups. Blood urea nitrogen (BUN) and creatinine levels dropped in NACNP compared with the PQ group and the NAC. Histological studies also approved PQ-induced damage and the protective effect of NACNP. The results indicated that NACNP could modulate oxidative stress status and kidney function against PQ toxicity.


22 week assessment of bladder acellular matrix as a bladder augmentation material in a porcine model.
Previous studies on the reconstruction of porcine bladder using bladder acellular matrix allograft (BAMA) have indicated positive preliminary results with respect to graft shrinkage and cellular repopulation. The current study was conducted to investigate the feasibility of using BAMA in a similar model of bladder reconstruction out to longer time frames (22 weeks). At predetermined time points, the macroscopic, histological and mechanical properties of explanted native and BAMA tissues were evaluated and compared. Macroscopically, contracture of the BAMA was observed. The peripheral regions of the grafts experienced extensive cellular repopulation. Towards the centre however, all grafts were consistently devoid of organized smooth muscle bundles and a well-developed urothelium. An alteration in both the amount and organization of collagen was also observed within this region. Significant differences (p < 0.05) in the rupture strain and the elastic modulus of the BAMA compared to native bladder tissue appear to correlate with macroscopic graft contracture as well as the fibroproliferative tissue response of the matrix.


Pacemaker utilization during permanent atrial fibrillation in patients who received pacemaker implantation for sinus node dysfunction.
Patients who have pacemakers and sinus node dysfunction frequently have atrial fibrillation (AF). The need for continued pacemaker therapy after conversion to permanent AF remains uncertain. This study showed that, among 174 patients who received pacemaker implantation for sinus node dysfunction, 38% (n = 62) had the minimum intrinsic ventricular rate of >60 beats/min after conversion to AF. The pacemaker memory showed that 30 patients (18%) never used ventricular pacing during permanent AF. The study results suggest that patients who have a stable intrinsic ventricular rate during permanent AF by serial assessment may no longer need continued pacemaker therapy.


Bystander effect induced by counted high-LET particles in confluent human fibroblasts: a mechanistic study.
The possible mechanism of a radiation-induced bystander response was investigated by using a high-LET heavy particle microbeam, which allows selected cells to be individually hit with precise numbered particles. Even when only a single cell within the confluent culture was hit by one particle of 40Ar (approximately 1260 keV/microm) or 20Ne (approximately 380 keV/microm), a 1.4-fold increase of micronuclei (MN) was detected demonstrating a bystander response. When the number of targeted cells increased, the number of MN biphasically increased; however, the efficiency of MN induction per targeted cell markedly decreased. When 49 cells in the culture were individually hit by 1 to 4 particles, the production of MN in the irradiated cultures were approximately 2-fold higher than control levels but independent of the number and LET of the particles. MN induction in the irradiated-culture was partly reduced by treatment with DMSO, a scavenger of reactive oxygen species (ROS), and was almost fully suppressed by the mixture of DMSO and PMA, an inhibitor of gap junctional intercellular communication (GJIC). Accordingly, both ROS and GJIC contribute to the above-mentioned bystander response and GJIC may play an essential role by mediating the release of soluble biochemical factors from targeted cells.


The impact of routine pulse oximetry use on outcomes in COVID-19-infected patients at increased risk of severe disease: A retrospective cohort analysis.
The phenomenon of silent hypoxaemia has been described in patients with COVID-19 pneumonia, which is characterised by low oxygen saturation levels of <90% in those who appear clinically well and do not show signs of significant respiratory distress. To assess the impact on clinical outcomes for high-risk COVID-19 patients using a pulse oximeter to monitor oxygen saturation levels in a home setting. We performed a retrospective cohort analysis using data from a large South African insurance administrator. Patients were categorised as high risk, based on age and specific underlying clinical conditions, or from predictive models derived from medical scheme administrative claims data. The impact of pulse oximetry home monitoring on COVID-19 clinical outcomes was investigated by the use of Cox proportional hazard models. Between 2 March 2020 and 31 October 2020, of 38 660 patients analysed, 8 115 were in the intervention group. The 60-day mortality rate for the evaluated high-risk population was 1.35%. After adjusting for age and comorbidity differences, the intervention group was found to have an adjusted hazard ratio of 0.52 (p<0.0001). No statistical significance was found between the intervened and control groups for admission to hospital, admission to intensive care unit (ICU) and use of mechanical ventilation. The intervention group had a lower median C-reactive protein (CRP) level on admission (p=0.03). After adjustment for admission CRP levels, elevated CRP was associated with an increased mortality (p<0.0001), while the statistical significance in mortality between the intervention and the control group was lost. High-risk COVID-19 patients who used a pulse oximeter to monitor oxygen saturation levels had significantly lower mortality rates compared with other high-risk patients. The mortality benefit may be explained by earlier presentation to hospital, as suggested by lower initial CRP levels.


Alterations in the field stimulation-induced release of endogenous norepinephrine from the coccygeal artery of spontaneously hypertensive and Wistar-Kyoto rats.
The field stimulation-induced release of endogenous NE from the isolated caudal artery from 5-6, 8-10 and 28-30 week old SHR resulted in a greater release of transmitter compared to age-matched WKY. The alpha 2-selective adrenoceptor antagonist, yohimbine produced a significant enhancement in the release of NE from both SHR and WKY of 5-6 and 10-12 and 28 week old WKY but release was attenuated in 28 week old SHR. It is concluded that the enhanced release of NE contributes to the development of hypertension in the SHR.


Cirrhotic livers reveal genetic changes in the MDM2-P14ARF system of cell cycle regulators.
The genesis of hepatocellular carcinoma is promoted by changes in the regulatory MDM2-P14ARF system. The incidence of such changes has to date not been analysed in non-tumourous livers showing regenerative proliferation. In the present study, 24 cirrhotic livers of alcohol-, autoimmue disorder- or HCV-caused genesis were screened for MDM2-P14ARF alterations at the level of protein, DNA and mRNA. Using confocal laser scanning microscopy, the absence of MDM2 and P14ARF expression was detected in all samples except three HCV-infected livers (four livers) which contained hepatocytes overexpressing MDM2 (P14ARF) protein. In two of the samples lacking P14ARF expression, laser microdissection and PCR demonstrated deletion of the P14ARF gene. The P14ARF gene amplified from other specimens did not carry mutations. MDM2 splicing variants were present in tissues from alcohol- and autoimmune disorder-induced cirrhoses. Sequencing of full-size mRNA revealed a MDM2 mis-sense mutation in an alcohol-induced cirrhosis. One sample contained regenerative nodules with genetic instability occurring at MDM2 locus D12S83 according to the data of automatic PCR fragment analysis. In summary, this study gives first evidence for different types of MDM2 and P14ARF alterations in cirrhotic livers. We suggest that the changes impair the regulatory MDM2-P14ARF system, thus possibly favouring regenerative proliferation and transformation.


Efficacy of light based detection systems for early detection of oral cancer and oral potentially malignant disorders: Systematic review.
Earlier detection of oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMD) is essential for dental professionals to improve patient survival rates. The aim of this systematic review is to to evaluate the effectiveness of devices that utilise the principles of chemiluminescence and tissue autofluorescence as adjuncts in the detection of OSCC and OPMD. The electronic retrieval systems and databases searched for relevant articles were PubMed [MEDLINE] and Science direct. The search was for limited articles published in English or with an English abstract and articles published during the period from January 2005 to April 2014. Clinical trials utilized ViziLite, Microlux TM/DL and Visual Enhanced Light scope (VELscope) for early detection of OPMD and OSCC. Twenty primary studies published satisfied our criteria for selection - 10 utilised chemiluminescence and 10 tissue autofluorescence. Senstivity of Vizilite for detecting OSCC nad OPMD ranged from 77.1 % to 100% and specificity was low that ranged from 0% to 27.8%.Most have shown that chemiluminescence increases the brightness and margins of oral mucosal white lesions and thus assist in identification of mucosal lesions not considered under Conventional visual examination. However, it preferentially detects leukoplakia and may fail to spot red patches. Clinical trials demonstrated that sensitivity of VELscope in detecting malignancy and OPMD ranged from 22 % to 100 % and specificity ranged from 16 % to 100%. Most studies concluded that VELscope can help the experienced clinician to find oral precursor malignant lesions. But it couldnot differentiate between dysplasia and benign inflammatory conditions. Both devices are simple, non-invasive test of the oral mucosa but are suited for clinicians with sufficient experience and training. More clinical trials in future should be conducted to establish optical imaging as an efficacious adjunct tool in early diagnosis of OSCC and OPMD.


Accurate sampling of PCDD/F in high temperature flue-gas using cooled sampling probes.
In a laboratory-scale combustion reactor, flue-gas samples were collected at two temperatures in the post-combustion zone, 700°C and 400°C, using two different water-cooled sampling probes. The probes were the cooled probe described in the European Standard method EN-1948:1, referred to as the original probe, and a modified probe that contained a salt/ice mixture to assist the cooling, referred to as the sub-zero probe. To determine the efficiency of the cooling probes, internal temperature measurements were recorded at 5cm intervals inside the probes. Flue-gas samples were analyzed for polychlorinated dibenzo-p-dioxin and dibenzofurans (PCDD/Fs). Samples collected at 700°C using the original cooling probe showed higher concentrations of PCDD/Fs compared to samples collected using the sub-zero probe. No significant differences were observed between samples collected at 400°C. The results indicated that artifact formation of PCDD/Fs readily occurs during flue-gas sampling at high temperatures if the cooling within the probe is insufficient, as found for the original probe at 700°C. It was also shown that this problem could be alleviated by using probes with an enhanced cooling capacity, such as the sub-zero probe. Although this may not affect samples collected for regulatory purposes in exit gases, it is of great importance for research conducted in the high-temperature region of the post-combustion zone.


Protein kinase of bacteriophage T7. 2. Properties, enzyme synthesis in vitro and regulation of enzyme synthesis and activity in vivo.
Protein kinase, which was isolated from cells infected with T7, is indeed a viral gene product. This is shown by DNA-dependent synthesis in vitro. The protein kinase transfers phosphate from ATP to seryl or threonyl residues in protein. The enzyme has only a relative requirement for magnesium ions, but is only active at low ionic strength. The best substrate is lysozyme. T7 protein kinase activity is not stimulated by cyclic 3':5'-AMP and/or cyclic 3':5'-GMP. The T7 protein kinase carries -- SH groups essential for activity. There is indication that the enzyme phosphorylates itself and causes self inactivation, which may explain the fast disappearance of enzyme activity in vivo. Bacteriophage T3 also induces a protein kinase which is similar to the T7-induced enzyme in all respects tested.


Association between current perceived stress and incident diabetes is dependent on occupational status: Evidence from the IPC cohort study.
The role of stress in the onset of type 2 diabetes is a widespread lay belief, yet observational studies have produced inconsistent results. This study aimed to test the hypothesis that the association between perceived stress and incident diabetes might depend on occupational status (OS). The four-item Perceived Stress Scale (PSS-4) was completed at baseline by 22,567 participants in the labour force (16,193 men, 6374 women; mean age: 44.5±9.8 years) who had undergone two health checkups subsidized by the French national healthcare system. All subjects were free from diabetes at baseline, defined as a fasting blood glycaemia≥7mmol/L or the use of antidiabetic drugs. After a mean follow-up of 5.3±2.1 years, 527 participants (2.3%) had incident diabetes. After adjusting for sociodemographic, behavioural and biomedical risk factors as well as self-rated health, the association between baseline perceived stress and diabetes at follow-up was non-significant for the total study population. However, perceived stress was significantly associated with incident diabetes in participants of low OS [odds ratio (OR) for a five-point increment: 1.39; 95% confidence interval (CI): 1.02-1.90]. In contrast, there was a negative association between perceived stress and diabetes among those of high OS (OR: 0.60; 95% CI: 0.41-0.88) and no association within other occupational categories. The interaction between perceived stress and OS was significant (P<0.01). This study suggests that the association between perceived stress and diabetes onset is dependent on OS. Furthermore, this association does not appear to be explained by the classical risk factors for diabetes.


Scale-up of Artemisia annua L. hairy root cultures produces complex patterns of terpenoid gene expression.
Hairy roots grow quickly, reach high densities, and can produce significant amounts of secondary metabolites, yet their scale-up to bioreactors remains challenging. Artemisia annua produces a rich array of terpenoids, including the sesquiterpene, artemisinin, and transformed roots of this species provide a good model for studying terpenoid production. These cultures were examined in shake flasks and compared with cultures grown in two types of bioreactors, a mist reactor and a bubble column reactor, which provide very different environments for the growing roots. Mist reactors have been shown previously to result in cultures that produce significantly more artemisinin per gram fresh weight of culture, while bubble column reactors have produced greater biomass. We have compared expression levels of four key terpenoid biosynthetic genes: 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), 1-deoxy-D-xylulose-5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), and farnesyl diphosphate synthase (FPS) in the three culture conditions. In shake flasks we found that although all four genes showed temporal regulation, only FPS expression correlated with artemisinin production. Light also affected the transcription of all four genes. Although expression in reactors was equivalent to or greater than that of roots grown in shake flasks, no correlation was found between expression level within six different zones of each reactor and their respective oxygen levels, light, and root-packing density. Surprisingly, transcriptional regulation of HMGR, DXS, DXR, and FPS was greatly affected by the position of the roots in each reactor. Thus, relying on a single reactor sample to characterize the gene activity in a whole reactor can be misleading, especially if the goal is to examine the difference between reactor types or operating parameters, steps essential in scaling up cultures for production.


Nanocomposite scaffold seeded with mesenchymal stem cells for bone repair.
The mechanical property of bone tissue scaffolds is one of the most important aspects in bone tissue engineering that has remained problematic. In our previous study, we fabricated a three-dimensional scaffold from nano-hydroxyapatite/gelatin (nHA/Gel) and investigated its efficiency in promoting bone regeneration both in vitro and in vivo. In the present study, the effect of adding silicon carbide (SiC) on the mechanical and biological behaviors of the nHA/Gel/SiC and bone regeneration in vivo were determined. nHA and SiC were synthesized and characterized by the X-ray diffraction pattern and transmission electron microscope image. Layer solvent casting, freeze drying, and lamination techniques were applied to prepare these scaffolds. Then, the biocompatibility and cell adhesion behavior of the synthesized nHA/Gel/SiC scaffolds were investigated. For in vivo studies, rats were categorized into three groups: blank defect, blank scaffold, and rat bone marrow mesenchymal stem cells (rBM-MSCs)/scaffold. After 1, 4, and 12 weeks post-injury, the rats were sacrificed and the calvaria were harvested. Sections with a thickness of 5 µm thickness were prepared and stained with hematoxylin-eosin and Masson's Trichrome, and immunohistochemistry was performed. Our results showed that SiC effectively increased the mechanical properties of the nHA/Gel/SiC scaffold. No significant differences were observed in biocompatibility, cell adhesion, and cytotoxicity of the nHA/Gel/SiC in comparison with the nHA/Gel nanocomposite. Based on histological and immunohistochemical studies, both osteogenesis and collagenization were significantly higher in the rBM-MSCs/scaffold group, quantitatively and qualitatively. The present study strongly suggests the potential of SiC as an alternative strategy to improve the mechanical and biological properties of bone tissue engineering scaffolds, and shows that the pre-seeded nHA/Gel/SiC scaffold with rBM-MSCs improves osteogenesis in the engineered bone implant.


Insecticidal properties and microbial contaminants in a Spodoptera exigua multiple nucleopolyhedrovirus (Baculoviridae) formulation stored at different temperatures.
The Spodoptera exigua (Hübner) multiple nucleopolyhedrovirus (SeMNPV) is currently being tested as a biological insecticide for use in greenhouse crops in southern Spain. We performed a study in which semipurified SeMNPV occlusion bodies (OBs) were formulated in phosphate-buffered saline, pH 6.5, with 5% (vol:vol) glycerol and 0.15% (wt:vol) sorbic acid, and they were stored at -20, 4, or 25 degrees C during 18 mo. Initial aerobic counts (+/-SE) averaged 1.4 (+/-0.17) x 10(7) colony-forming units/ml after 17-h incubation at 37 degrees C. Aerobic counts of microorganisms that contaminated OB formulations stored at 250C decreased markedly over the period of the study, whereas only small decreases were observed in counts from OBs stored at 4 or -20 degrees C. The principal microbial contaminants of OB suspensions were Enterococcus spp., Enterobacteriaceae, and yeasts. Potential human pathogens (Salmonella, Shigella, and Vibrio species) were not detected, and populations of Staphylococcus aureus and Bacillus cereus were extremely low. Compared with newly formulated OBs, the estimated LD50 values of OBs stored at 25 degrees C increased by >16,666-fold over the 18 mo of storage, whereas LD50 values were not greatly affected by storage at 4 or -20 degrees C. Significant changes over time in OB concentrations were only observed in the 25 degrees C treatment. Complete degradation of viral DNA was observed at 25 degrees C but not in refrigerated or frozen OBs. We conclude that OB formulation with bacteriostatic or antioxidant additives, together with storage and distribution in refrigerated conditions, will likely result in an SeMNPV biopesticide shelf life that exceeds 18 mo.


Medication-related osteonecrosis of the jaw without osteolysis on computed tomography: a retrospective and observational study.
Medication-related osteonecrosis of the jaw (MRONJ) is a refractory osteonecrosis caused by antiresorptive agents such as bisphosphonate and denosumab (DMB). In MRONJ surgery, computed tomography (CT) is generally used to determine the extent of bone resection. However, in some recent MRONJ cases, no abnormal findings were detected on CT. Therefore, we aimed to clarify the characteristics of MRONJ without osteolysis. This retrospective and observational study included 18 patients diagnosed with MRONJ between October 2020 and October 2022 at Department of Dentistry and Oral Surgery, Kansai Medical University Medical Center. In four of 18 patients with MRONJ, no abnormal findings such as osteolysis, separation of sequestrum, and periosteal reaction were observed on CT images at the first visit. All four patients with non-osteolytic MRONJ had malignant tumors and received high-dose DMB, and in the four patients there were no preceding dental infections such as apical lesions or periodontal disease and the trigger of MRONJ was unknown. Surgery was performed in three of the four patients. The extent of bone resection was determined using magnetic resonance imaging and intraoperative gross findings. In the future, it is necessary to establish a method for diagnosing non osteolytic MRONJ and determining the extent of bone resection.


Allocation of federal funding to address the opioid overdose crisis in the criminal legal system.
The opioid overdose crisis remains a chief public health concern in the United States, and people involved in the criminal legal system are among the most vulnerable to opioid related harms. This study aimed to identify all discretionary federal funding allocated to states, cities, and counties targeting the overdose crisis for criminal legal system-involved populations in fiscal year (FY) 2019. We then aimed to assess the extent to which federal funding was allocated to states with the highest need. We collected data from publicly available government databases (N = 22) to identify federal funding targeting opioid use disorder in criminal legal system-involved populations. Descriptive analyses examined the extent to which funding allocated per person in the criminal legal system-involved population was associated with funding need, proxied by a composite measure of opioid mortality and drug-related arrests. We created a generosity measure and dissimilarity index to assess the degree to which funding matched need across states. More than 590 million dollars were allocated across 517 grants by 10 federal agencies in FY 2019. About half of states received less than $100.00 dollars per capita in the state criminal legal system-involved population. Funding generosity ranged from 0 % to 504.2 %, with more than half of states (52.9 %, n = 27) receiving fewer dollars per opioid problem than the US average. Further, a dissimilarity index indicated that about 34.2 % of funding (~$202.3 million) would have to be reallocated to distribute funding more evenly across states. Results suggest that additional efforts are needed to more equitably distribute funds to meet the needs of states with more severe opioid problems.


Changes in the number of lower limb amputations during a period of increasing vascular surgical activity. Results of a nation-wide study, Denmark, 1977-1990.
To find out if the number of lower limb amputations for peripheral occlusive arterial disease increased during the period of increased vascular surgical activity 1977-1990. Retrospective review of a database covering all admissions to Danish hospitals. All patients admitted to Danish hospitals with peripheral occlusive disease during the period 1977-1990. Numbers, and age and sex specific incidence rates for admission, arterial reconstruction and amputation. The number of admissions during which arterial reconstruction for OAD was done increased from 963 in 1977 to 2311 in 1990 (p < 0.001). The number of admissions during which a diagnosis of OAD was made and an amputation done increased from 1290 in 1977 to a maximum of 1609 in 1983 (p < 0.001). The number remained nearly constant in the mid-1980s but decreased during last part of the period to 1181 in 1990 (p < 0.001). The reduction in the number of amputations may indicate that vascular surgery even on a national scale is effective in preventing amputations for OAD.


Biosynthesis of GDP-fucose and other sugar nucleotides in the blood stages of Plasmodium falciparum.
Carbohydrate structures play important roles in many biological processes, including cell adhesion, cell-cell communication, and host-pathogen interactions. Sugar nucleotides are activated forms of sugars used by the cell as donors for most glycosylation reactions. Using a liquid chromatography-tandem mass spectrometry-based method, we identified and quantified the pools of UDP-glucose, UDP-galactose, UDP-N-acetylglucosamine, GDP-mannose, and GDP-fucose in Plasmodium falciparum intraerythrocytic life stages. We assembled these data with the in silico functional reconstruction of the parasite metabolic pathways obtained from the P. falciparum annotated genome, exposing new active biosynthetic routes crucial for further glycosylation reactions. Fucose is a sugar present in glycoconjugates often associated with recognition and adhesion events. Thus, the GDP-fucose precursor is essential in a wide variety of organisms. P. falciparum presents homologues of GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase enzymes that are active in vitro, indicating that most GDP-fucose is formed by a de novo pathway that involves the bioconversion of GDP-mannose. Homologues for enzymes involved in a fucose salvage pathway are apparently absent in the P. falciparum genome. This is in agreement with in vivo metabolic labeling experiments showing that fucose is not significantly incorporated by the parasite. Fluorescence microscopy of epitope-tagged versions of P. falciparum GDP-mannose 4,6-dehydratase and GDP-L-fucose synthase expressed in transgenic 3D7 parasites shows that these enzymes localize in the cytoplasm of P. falciparum during the intraerythrocytic developmental cycle. Although the function of fucose in the parasite is not known, the presence of GDP-fucose suggests that the metabolite may be used for further fucosylation reactions.


Cytosines do it, thymines do it, even pseudouridines do it--base flipping by an enzyme that acts on RNA.
In the December 28, 2001 issue of Cell, Hoang and Ferré-D'Amaré report the structure of a tRNA pseudouridine synthase, showing the target uridine flipped out from the tRNA and confirming that base flipping is not limited to enzymes that act on DNA.


James A. Rand Young Investigator's Award: Questioning the "Nickel Free" Total Knee Arthroplasty.
A study was performed to measure metal ions present in the knee joint after performing a total knee arthroplasty (TKA) with standard cobalt chromium (CoCr) components as well as with "nickel-free" oxidized zirconium femoral and titanium tibial (OxZr/Ti) components. Knee joint fluid was collected prior to arthrotomy, and on postoperative day one to determine the amount of metal debris generated when performing a TKA with standard instrumentation from consecutive cases with CoCr components (n = 24) and OxZr/Ti components (n = 16). CoCr implant patients had statistically higher levels of nickel (Ni) (29.7%, P = .033), cobalt (Co), (1,100.7%, P < .0001) and chromium (Cr) (118.9%, P < .0001) postoperatively. The cutting blocks and sawblades do not contain Co, which therefore must have come from the components. The metal ions generated from the sawblades and cutting blocks, therefore, could be discerned from the OxZr/Ti whose components don't contain Co, Cr, or Ni. The OxZr patients had significantly higher Cr (9.5×, P < .001) and Ni (5.1×, P < .001) post-TKA vs pre-TKA; Co levels were not significantly different as expected with the absence of Co in the components (P = .60). The Ni levels generated in performing an Oxinium TKA was 3.3 times higher than when performing a CoCr TKA (1.37 vs. 41 ppb, P < .001). The substantial degree of Ni generation resulting from performing a hypoallergenic "nickel-free" TKA calls into questions the rationale of utilizing more expensive lower Ni components on the basis of known or suspected Ni or Cr allergy.


A conjugative plasmid pTE195 coding for drug resistance and virulence phenotypes from Salmonella naestved strain of calf origin.
Salmonella naestved strain AHI-195, of calf origin, harbors a conjugative 95 megadalton (MDa) plasmid, pTE195, which encodes resistance to tetracycline and chloramphenicol and belongs to incompatibility group FII. Moreover, DNA homology between pTE195 and the Salmonella dublin virulence plasmid pTE800 was revealed by digestion with several restriction endonucleases and confirmed by hybridization with different specific probes. These results indicate that pTE195 carries not only genes for drug resistance but also genes for virulence phenotypes such as serum resistance and mouse lethality.


Ambulation and Parkinson disease.
Parkinson disease is a progressive neurodegenerative disorder characterized by a variety of motor and nonmotor features. This article reviews the problems of postural instability and gait disturbance in persons with Parkinson disease through the discussion of (1) the neuropathology of parkinsonian motor deficits, (2) behavioral manifestations of gait and postural abnormalities observed in persons with Parkinson disease, and (3) pharmacologic, surgical, and physical therapy-based interventions to combat postural instability and gait disturbance. This article advances the treatment of postural instability and gait disturbance by condensing up-to-date knowledge and making it available to clinicians and rehabilitation professionals.


Morphology and expression status investigations of specific surface markers on B-cell chronic lymphocytic leukemia cells.
The morphology of cells and expression status of specific surface markers [cluster of differentiation (CD)], such as CD5, CD19, CD20, CD38, and CD45, have long been considered as the essential indicators for the diagnosis and prognosis of B-cell chronic lymphocytic leukemia (B-CLL). Clinically, it is difficult to simultaneously obtain cell morphology and distribution of surface markers with flow cytometry, especially for some surrogate markers such as CD38. Here, as an alternative and complementary prognostic method, fluorescence microscopy and image processing method are introduced to directly visualize the cells from patients and to quantitatively determine the expression status of surface markers. In this study, the morphological parameters of B-CLL cells were measured to establish the correlation between the cellular morphology and the surface marker expression. It was clear that the CD38+ and CD38- B-CLL cells from the same CD38+ patients had hardly any size differences; however, an increase in perimeter was observed for CD38- patients. Moreover, the expression level of the receptors on the cell was independent of the cell size. There was no evidence showing that the expression intensities of CD19 and CD38 were related to each other for the CD38+ B-CLL cells. On the same cells, CD5 was more selectively expressed on the cell membrane; however, the expression patterns suggested that the cell membrane of CD38- B-CLL cells contained the least expression level of CD19.


The mechanism and clinical implication of improved left ventricular videointensity following intravenous injection of multi-fold dilutions of albumin with dextrose.
The left ventricular ultrasound videointensity of an intravenous injection of sonicated albumin is improved if the agent is diluted with dextrose prior to sonication. The objective of this study was to determine the mechanism for improved left ventricular ultrasound contrast with intravenous sonicated multi-fold dilutions of albumin with dextrose compared to sonicated albumin alone. Epicardial short axis images of the left ventricle were obtained in 11 mongrel dogs and incremental one part sonicated dilutions (up to 10-fold) of albumin with 5 or 50% dextrose were given intravenously to determine which dilution and dextrose concentration produced optimal left ventricular videointensity. Microbubble size and concentration of these dilutions were measured. The one to seven-fold sonicated dilutions resulted in a slight, but significantly larger microbubble size when compared to sonicated albumin alone (SA), but no difference in concentration. All dilutions produced significantly higher end-diastolic peak videointensity (PVI) in the left ventricle than SA (range 160-569% of SA PVI; p < 0.001) with the three to five-fold dilution producing maximal PVI. Five percent dextrose dilutions produced the same videointensity as 50% dilutions. End-systolic videointensity of both 5 and 50% dextrose dilutions were also over 250% higher than SA (p < 0.001). This resulted in good or excellent end-systolic endocardial border definition in the majority of injections. Therefore, the mechanism for improved left ventricular chamber opacification with multifold sonicated dilutions of albumin with dextrose appears to be due to a small increase in microbubble size and not increased viscosity or microbubble concentration.


Uncertainty aware training to improve deep learning model calibration for classification of cardiac MR images.
Quantifying uncertainty of predictions has been identified as one way to develop more trustworthy artificial intelligence (AI) models beyond conventional reporting of performance metrics. When considering their role in a clinical decision support setting, AI classification models should ideally avoid confident wrong predictions and maximise the confidence of correct predictions. Models that do this are said to be well calibrated with regard to confidence. However, relatively little attention has been paid to how to improve calibration when training these models, i.e. to make the training strategy uncertainty-aware. In this work we: (i) evaluate three novel uncertainty-aware training strategies with regard to a range of accuracy and calibration performance measures, comparing against two state-of-the-art approaches, (ii) quantify the data (aleatoric) and model (epistemic) uncertainty of all models and (iii) evaluate the impact of using a model calibration measure for model selection in uncertainty-aware training, in contrast to the normal accuracy-based measures. We perform our analysis using two different clinical applications: cardiac resynchronisation therapy (CRT) response prediction and coronary artery disease (CAD) diagnosis from cardiac magnetic resonance (CMR) images. The best-performing model in terms of both classification accuracy and the most common calibration measure, expected calibration error (ECE) was the Confidence Weight method, a novel approach that weights the loss of samples to explicitly penalise confident incorrect predictions. The method reduced the ECE by 17% for CRT response prediction and by 22% for CAD diagnosis when compared to a baseline classifier in which no uncertainty-aware strategy was included. In both applications, as well as reducing the ECE there was a slight increase in accuracy from 69% to 70% and 70% to 72% for CRT response prediction and CAD diagnosis respectively. However, our analysis showed a lack of consistency in terms of optimal models when using different calibration measures. This indicates the need for careful consideration of performance metrics when training and selecting models for complex high risk applications in healthcare.


Hemodynamic management of the micropreemie: When inotropes are not enough.
Managing perfusion in the micropreemie is challenging and should be guided by the patho-physiology, gestational and postnatal age of the baby, perinatal history, and the persistence of fetal shunts. The assessment should incorporate bedside tools such as blood pressure, clinical perfusion markers, and functional echocardiography. The multimodal approach to diagnose and identify the cause of hemodynamic compromise paves the way to a targeted approach to treatment. Characterizing the predominant pathophysiologic cause of low cardiac output and impaired cellular metabolism enables a more accurate use of inotropes, vasopressors, and volume support to suit a particular pathophysiologic situation.


Retromer.
The retromer is a heteropentameric complex that associates with the cytosolic face of endosomes and mediates retrograde transport of transmembrane cargo from endosomes to the trans-Golgi network. The mammalian retromer complex comprises a sorting nexin dimer composed of a still undefined combination of SNX1, SNX2, SNX5 and SNX6, and a cargo-recognition trimer composed of Vps26, Vps29 and Vps35. The SNX subunits contain PX and BAR domains that allow binding to PI(3)P enriched, highly curved membranes of endosomal vesicles and tubules, while Vps26, Vps29 and Vps35 have arrestin, phosphoesterase and alpha-solenoid folds, respectively. Recent studies have implicated retromer in a broad range of physiological, developmental and pathological processes, underscoring the critical nature of retrograde transport mediated by this complex.


[Corrigendum] Anti‑osteoporotic effects of tetramethylpyrazine via promoting osteogenic differentiation and inhibiting osteoclast formation.
Subsequently to the publication of the above paper, an interested reader drew to the authors' attention that, in Fig. 2A on p. 8311, portraying the results of immunostaining experiments for osterix, the 'GIOP' and 'GIOP+TMP (20)' data panels contained overlapping data, such that these images were derived from apparently the same original source, where they were intended to show the results from differently performed experiments. Moreover, in Fig. 3A on p. 8312 showing the results from ALP staining and Alizarin Red S staining experiments, two pairs of apparently overlapping data panels were identified in the Dex 106 M / TMP 50 μM, 100 μM and 200 μM data panels. After having re‑examined their original data, the authors have realized that the data featured in Figs. 2A and 3A were assembled incorrectly in these figures. Revised versions of Fig. 2 and 3, now containing replacement data for the experiments shown in Figs. 2A and 3A, are shown on the next page. Note that these errors did not adversely affect either the results or the overall conclusions reported in this study. All the authors agree with the publication of this corrigendum, and are grateful to the Editor of Molecular Medicine Reports for allowing them the opportunity to publish this. They also wish to apologize to the readership of the Journal for any inconvenience caused. [Molecular Medicine Reports 16: 8307‑8314, 2017; DOI: 10.3892/mmr.2017.7610].


Multiple skeletal anomalies in the "13q-" syndrome.
A patient with the "13q-" syndrome is reported. The typical association of congenital malformations was found. Multiple and unusual skeletal anomalies included absent thumbs, club-feet, coxa vara, diastasis of the pubic symphisis and extensive spina bifida occulta. These appear to be part of the multiple system involvement due to the chromosomal deletion. Chromosome analysis is indicated in patients with multiple skeletal anomalies, especially if the thumbs and radial axis are involved.


Attributional style among youth at clinical risk for psychosis.
A biased attributional style, in which negative events are attributed to external and personal causes, is associated with paranoid delusions in schizophrenia. It is not known whether this biased attributional style also characterizes individuals at clinical risk for psychosis or if it is associated with their emergent paranoia. Thirty-three clinical high-risk patients and 15 age- and gender-similar controls were assessed with the Internal, Personal, and Situational Attributions Questionnaire for externalizing and personalizing attributional biases and for potential correlates with suspiciousness and other symptoms. Both patients and controls had a similar external-personalizing attributional style that was unrelated to symptoms, including suspiciousness. Consistent with other studies, a biased attributional style was not associated with subthreshold paranoia. Therefore, a biased attributional style is likely not a trait that contributes to emergent paranoid delusions but is instead a state-dependent correlate of paranoid delusions.


Cytotoxicity of commercial nano-TiO2 to Escherichia coli assessed by high-throughput screening: effects of environmental factors.
The extensive use of nano-TiO2 in industry has led to growing concerns about its potential environmental impacts. However, negligible toxicity is commonly reported under insufficient illumination and artificial solution conditions in the literature, which rarely includes discussion of the regulating role of environmental factors. Herein, we report the results of a high-throughput screening assay to evaluate the acute cytotoxicity of six commercial nano-TiO2 materials to Escherichia coli (E. coli) using Lake Michigan water as a model for aquatic surface environments. In particular, we investigate the specific effects of illumination wavelength and natural organic matter (NOM) content. Under simulated solar irradiation, four anatase-based nano-TiO2 materials including Pigment White 6 exhibit significant bacterial toxicity (2 h-IC50 value of 2.7-9.1 mg/L), with toxicity thresholds much lower than previously reported. Negligible toxicity is caused either by pure-phase rutile or under dark condition. Formation of nano-TiO2 aggregates well beyond nano-scale does not eliminate their toxic effect, but photoactivity dominates over the primary size and extent of aggregation in determining the acute cytotoxicity of nano-TiO2. Under visible light irradiation (UVA&B blocked) the antibacterial activity of nano-TiO2 is essentially erased, whereas removing only UVB wavelengths slightly mitigates the toxicity. Suwannee River fulvic acid, acting as a natural dispersant, reverses the extent of nano-TiO2 aggregation, but also reduces its bacterial cytotoxicity. These results demonstrate that despite particle aggregation, the short-term cytotoxicity of nano-TiO2 is predominantly attributed to its phototoxicity, emphasizing the importance of illumination conditions in toxicological screening of photoactive nanomaterials. In the natural aquatic environment, however, this acute toxicity may be mitigated by the attenuation of UV irradiation and increased NOM concentration in the water column.


General overview of the use of computers in nursing research.
Carol Cooper offers some useful tips on using computer programs to enhance nursing research.


Akt1/protein kinase B enhances transcriptional reprogramming of fibroblasts to functional cardiomyocytes.
Conversion of fibroblasts to functional cardiomyocytes represents a potential approach for restoring cardiac function after myocardial injury, but the technique thus far has been slow and inefficient. To improve the efficiency of reprogramming fibroblasts to cardiac-like myocytes (iCMs) by cardiac transcription factors [Gata4, Hand2, Mef2c, and Tbx5 (GHMT)], we screened 192 protein kinases and discovered that Akt/protein kinase B dramatically accelerates and amplifies this process in three different types of fibroblasts (mouse embryo, adult cardiac, and tail tip). Approximately 50% of reprogrammed mouse embryo fibroblasts displayed spontaneous beating after 3 wk of induction by Akt plus GHMT. Furthermore, addition of Akt1 to GHMT evoked a more mature cardiac phenotype for iCMs, as seen by enhanced polynucleation, cellular hypertrophy, gene expression, and metabolic reprogramming. Insulin-like growth factor 1 (IGF1) and phosphoinositol 3-kinase (PI3K) acted upstream of Akt whereas the mitochondrial target of rapamycin complex 1 (mTORC1) and forkhead box o3 (Foxo3a) acted downstream of Akt to influence fibroblast-to-cardiomyocyte reprogramming. These findings provide insights into the molecular basis of cardiac reprogramming and represent an important step toward further application of this technique.


Receptor tyrosine kinase alterations and therapeutic opportunities in squamous cell carcinoma of the lung.
Targeted therapy has greatly improved the treatment for adenocarcinoma of the lung, but not squamous cell carcinoma (SCC) of the lung. The current paper describes the abnormalities of receptor tyrosine kinases (RTK) in lung SCC in a hope to stimulate the development of therapeutics that can have clinical impact. We reviewed both clinical and preclinical studies published in English regarding RTK abnormalities and/ or RTK-targeting treatment for SCC of the lung. RTK alterations have been demonstrated as biological signature for SCC of the lung. A number of clinical trials of RTK-targeting therapy have been carried out or are ongoing, with encouraging results. SCC of the lung should be treated as an independent disease with unique treatment options based on molecular changes, particularly RTK.


Determination of focal ischemic lesion volume in the rat brain using multispectral analysis.
Multispectral (MS) analysis was used to determine the ischemic lesion volume in the rat brain after permanent middle cerebral arterial occlusion. MS analysis used a four-dimensional MS model consisting of an estimate of the average apparent diffusion coefficient of water (ADC(av)), T2, proton density, and perfusion. Four classification methods were investigated: (a) multivariate gaussian (MVG); (b) k-nearest neighbor (k-NN); (c) k-means (KM); and (d) fuzzy c-means (FCM). MVG and k-NN classifiers are supervised methods requiring labeled training data to characterize the stroke lesion. Unsupervised classifiers (KM, FCM) do not require previous statistics or labeled training data, resulting in potentially greater clinical usefulness. All MS methods provided significant correlation with postmortem findings beyond the use of ADC(av) alone (partial correlation given the ADC(av) estimate: MVG, .66; k-NN, .75; KM, .68; FCM, .70). This study demonstrates that MS analysis provides an improved estimate of ischemic lesion volume over that obtained from ADC alone.


Human Rhinovirus Infection Enhances the Th2 Environment in Allergic and Non-allergic Patients with Chronic Rhinosinusitis.
This study was conducted to determine whether patients with allergic rhinitis might be more susceptible to human rhinovirus (HRV) infection and whether the effects of infection on the elicited immune responses are different in allergic and non-allergic patients with chronic rhinosinusitis (CRS). Uncinate process tissues were obtained from 61 CRS patients (of whom 39 had allergies and 22 did not) and were infected with HRV-16 using an air-liquid interface organ culture system. The expression levels of programmed cell death-ligand (PD-L)1, PD-L2, intracellular adhesion molecule 1, interferon-gamma (IFN-γ), interleukin (IL)-4, IL-5, and IL-10 were evaluated in the infected nasal mucosa. The HRV infection rates were not significantly different between the allergy (74.4%) and non-allergy (72.7%) groups. In the allergy group, the expression of PD-L1 (P=0.013) and IL-10 (P=0.040) was significantly elevated in the HRV-infected tissues, and there was a strong correlation between PD-L1 and IL-10 (r=0.868, P<0.001). In contrast, infected tissues from the non-allergy group displayed increased levels of IL-4 (P=0.039), IL-5 (P=0.023), and IFN-γ (P=0.031), as well as an increased IL-4/IFN-γ ratio, after HRV infection (P=0.043). This study showed that HRV infection rates were similar in the nasal mucosa of patients with CRS regardless of the presence of allergic rhinitis. HRV infection enhanced the Th2 environment by modulating PD-L1 and PD-L2 expression levels in allergic mucosa and by increasing the IL-4/IFN-γ ratio in non-allergic mucosa.


The Hartmann procedure. Extended indications in severe intra-abdominal infection.
During a three-year period, 30 patients had emergency Hartmann procedures for diverticular disease (N = 12), carcinoma (N = 6), trauma (N = 3), and miscellaneous causes (N = 9). Two patients died postoperatively (6.7 percent) and wound infection developed in 60 percent of the patients. Planned relaparotomies for severe intra-abdominal infection were performed in ten patients (an average of 2.5 procedures per patient) with no mortality. In five cases a mucous fistula was converted into a Hartmann pouch; a preference for the Hartmann pouch in patients undergoing repeated explorations is discussed. Colorectal continuity was subsequently restored in 23 patients (76.6 percent).


Reduction in trabecular meshwork stem cell content in donor eyes with primary open angle glaucoma.
We previously identified and characterized human trabecular meshwork stem cells (TMSCs) based on high expression of ABCG2/p75 positivity and high nucleus to cytoplasmic ratio. These TMSCs expressing high ABCG2 and p75 were located to the insert region of the human TM. Additionally, we demonstrated an age-related reduction in the TMSC content which was significantly associated with TM cell loss. In continuation, this study was aimed to determine the TMSC content in glaucomatous donor eyes wherein a drastic reduction in TM cellularity has already been reported. Anterior segments from known glaucomatous (n = 6) and age-matched normal (n = 8) donors were dissected into four quadrants. A minimum of three sections from each quadrant were used for histopathological analysis as well as immunostaining. Analysis of hematoxylin and eosin-stained sections from glaucomatous tissues revealed a decrease in total TM cellularity, thickening of trabecular beams, fusion of trabeculae, absence of patent Schlemm's canal compared to age-matched controls. In addition, the TM thickness at various positions of the meshwork and the coronal as well as the meridional diameters of the Schlemm's canal were observed to be significantly reduced in glaucomatous eyes. Further, sections from both the groups were immunostained for universal stem cell marker ABCG2 and neural crest derived stem cell marker p75. The images were acquired using Leica SP8 confocal microscope. Quantification of total TM cellularity based on nuclear counterstain (mean ± SD) using ImageJ identified 69.33 ± 12.77 cells/section in control eyes. In glaucomatous donors, the TM cellularity was found to be reduced significantly to 41.83 ± 9.0 (p = 0.0007). In addition, a reduction in the percentage of TMSCs (cells with high ABCG2 expression and p75 positivity) was evident in glaucomatous donors (0.14 ± 0.17%) compared to age-matched controls (4.73 ± 5.46%) (p = 0.064). Thus, the present study confirmed the significant decline in TM cellularity and a reducing trend in the TMSC content, though this reduction was non-significant in glaucomatous donor eyes. Further studies are essential to elucidate the role of TMSCs in the pathogenesis of primary open angle glaucoma.


DNA damage and repair induced by diazoacetyl derivatives of amino acids with different mechanism of cytotoxicity. Correlations with mutagenicity and carcinogenicity.
Eight synthetic N-diazoacetyl amino acids, prepared by inserting a diazoacetyl group onto the alpha-nitrogen of a natural amino acid, and two natural diazoazetyl amino acids, azaserine (9-diazoacetyl-L-serine) and DON (6-diazo-5-oxo-L-norleucine), have been studied by autoradiography for their capacity to induce DNA repair synthesis in mouse cells cultivated "in vitro". Dose-dependent unscheduled DNA synthesis was present in cells treated with the eight N-diazoacetyl derivatives, and was absent in cells exposed to approximately equitoxic concentrations of azaserine and DON. Azaserine and DON, unlike N-diazoacetyl derivatives, did not alkylate gamma-(4-nitrobenzyl) pyridine at an appreciable extent. When DNA damage (single stranded breaks or weak points in alkali) was measured by the sensitive technique of alkaline elution, DGA was found about 4 times as potent as azaserine and about 12 times as DON on a molar basis, but about 800 and 17,000 times as potent as azaserine and DON respectively by extrapolating to equitoxic concentrations. Carcinogenicity and mutagenicity seem to follow mainly the capability of inducing DNA damage.


Chalcane-stilbene conjugates and oligomeric flavonoids from Chinese Dragon's Blood produced from Dracaena cochinchinensis.
A detailed chemical investigation of Chinese Dragon's Blood, which is a traditional medicine produced form the red resin of Dracaena cochinchinensis, yielded two chalcane-stilbene conjugates, named cochinchinenenes G and H, together with 25 known compounds. The structures of these compounds were determined by spectroscopic examination. HPLC analysis of the resin indicated that the major constituents were a complex mixture of oligomeric polyphenols, which were detected as a broad hump on the base line of a HPLC chromatogram. (13)C NMR analysis indicated that the oligomers were mainly composed of oxygenated chalcane units. This suggestion was supported by the results of a thiol degradation experiment with mercaptoethanol, which yielded a thioether of 4-[(4-hydroxyphenyl)propyl]-3-methoxyphenol. Furthermore, methylation followed by electrospray ionization mass spectroscopic analysis of the resulting fractions established the presence of at least one heptamer of chalcane units.


The measurement-based care to opioid treatment programs project (MBC2OTP): a study protocol using rapid assessment procedure informed clinical ethnography.
Psychosocial interventions are needed to enhance patient engagement and retention in medication treatment within opioid treatment programs. Measurement-based care (MBC), an evidence-based intervention structure that involves ongoing monitoring of treatment progress over time to assess the need for treatment modifications, has been recommended as a flexible and low-cost intervention for opioid treatment program use. The MBC2OTP Project is a two-phase pilot hybrid type 1 effectiveness-implementation trial that has three specific aims: (1) to employ Rapid Assessment Procedure Informed Clinical Ethnography (RAPICE) to collect mixed methods data to inform MBC implementation; (2) to use RAPICE data to adapt an MBC protocol; and (3) to conduct a hybrid type 1 trial to evaluate MBC's preliminary effectiveness and implementation potential in opioid treatment programs. This study will be conducted in two phases. Phase 1 will include RAPICE site visits, qualitative interviews (N = 32-48 total), and quantitative surveys (N = 64-80 total) with staff at eight programs to build community partnerships and evaluate contextual factors impacting MBC implementation. Mixed methods data will be analyzed using immersion/crystallization and thematic analysis to inform MBC adaptation and site selection. Four programs selected for Phase 2 will participate in MBC electronic medical record integration, training, and ongoing support. Chart reviews will be completed in the 6 months prior-to and following MBC integration (N = 160 charts, 80 pre and post) to evaluate effectiveness (patient opioid abstinence and treatment engagement) and implementation outcomes (counselor MBC exposure and fidelity). This study is among the first to take forward recommendations to implement and evaluate MBC in opioid treatment programs. It will also employ an innovative RAPICE approach to enhance the quality and rigor of data collection and inform the development of an MBC protocol best matched to opioid treatment programs. Overall, this work seeks to enhance treatment provision and clinical outcomes for patients with opioid use disorder. Trial registration This study will be registered with Clinicaltrials.gov within 21 days of first participant enrollment in Phase 2. Study Phase 1 (RAPICE) does not qualify as a clinical trial, therefore Phase 2 clinical trial registration has not yet been pursued because all elements of Phase 2 will be dependent on Phase 1 outcomes.


The association of predeployment and deployment-related factors on dimensions of postdeployment wellness in U.S. military service members.
To assess the effects of predeployment and deployment-related factors on dimensions of wellness following deployment. Prospective longitudinal study. The dependent variable was dimensions of wellness. Independent variables were measured in terms of modifiable, nonmodifiable, and military factors, such as sex, race/ethnicity, service branch, smoking status, and combat experience. A large military cohort participating in the Millennium Cohort Study. Included 10,228 participants who deployed in support of the operations in Iraq and Afghanistan. Dimensions of wellness were measured by using standardized instruments assessing self-reported physical health, mental health, and stress. Covariates were measured by using self-reported and electronic data. Factors of postdeployment wellness were assessed by using ordinal logistic regression. Most participants (78.7%) were categorized as "moderately well" post deployment. Significant modifiable predeployment predictors of postdeployment wellness included normal/underweight body mass index (odds ratio [OR] = 1.72, p < .05). Military factors significantly associated with wellness included not experiencing combat (OR = .56, p < .05), member of Air Force (OR = 2.02, p < .05) or Navy/Coast Guard (OR = 1.47, p < .05), and combat specialist occupation (OR = 1.22, p < .05). Multiple modifiable factors associated with postdeployment wellness were identified, which may help inform medical and military leadership on potential strategies to ensure a well force. Those trained in combat roles were more likely to be well post deployment though this apparent benefit was not conferred onto those reporting combat experiences.


Jejunal malignant lymphoma in a patient with adult-onset hypo-gamma-globulinemia and nodular lymphoid hyperplasia of the small bowel.
A malignant lymphoma of the small bowel developed in a 55-yr-old woman who had intestinal malabsorption for 20 yr, late-onset hypo-gamma-globulinemia, and nodular lymphoid hyperplasia. The tumor arose in the first jejunal loop and appeared to be of B-immunoblastic type (diffuse large noncleaved cells). This case indicates that nodular lymphoid hyperplasia in the context of immunodeficiency may be a condition leading to lymphoid malignancy.


Vitamin D and disability in relapsing-remitting multiple sclerosis in patients with a Mexican background.
Previous studies of multiple sclerosis (MS) patients have reported an inverse correlation between disability, the number of relapses and vitamin D levels in mostly white patients. It is unclear if this relationship has the same behavior in individuals with Hispanic backgrounds. To determine the relationship between vitamin D serum levels and disability in a sample of Hispanics of a Mexican background with relapsing-remitting multiple sclerosis (RRMS). A cross-sectional study was conducted on 50 RRMS individuals of Mexican background. The Expanded Disability Status Scale (EDSS) score, progression index (PI) and annual relapse rate (ARR) were recorded for each patient. Vitamin D levels were assessed during the summer. Pearson's test was used to evaluate the relationship between vitamin D and EDSS, PI, ARR, and duration of disease evolution. Most patients were females (n = 29, 58%). The mean vitamin D level was 22.3 (± 6.4) ng/ml; the mean EDSS score was 2.2 (± 0.7), ARR 1.3 (± 0.5) and PI1.08 (± 0.6). No correlation was found between vitamin D levels and EDSS scores, ARR, PI or duration of disease. Moderate negative association between vitamin D levels and EDSS was found just in females (<0.0001). No correlation between vitamin D levels and disability was found in this sample of RRMS Mexicans. Longitudinal studies are needed to better understand the impact of Vitamin D in disability and multiple time points.


The effect of human umbilical cord mesenchymal stem cell-derived exosomes on diabetic retinal neurodegeneration in a rat model.
To investigate the effect of human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Exs) on diabetic retinal neurodegeneration (DRN). Exosomes were isolated from human umbilical cord mesenchymal stem cells (hucMSC) and identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting (WB). Rats were intraperitoneally injected with Streptozotocin (STZ) to establish a diabetes mellitus model, and blood glucose levels and body weight were assessed. The rats were intravitreally injected with phosphate buffered saline (PBS; diabetic group) or hucMSC-Exs (hucMSC-Exs group). A control group of rats were not treated with STZ and were intravitreally injected with PBS (normal control group). Hematoxylin-eosin (HE) staining was used to observe changes in retinal structure and to count the number of retinal ganglion cells (RGCs) four weeks after intravitreal injection. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL) was used to detect retinal cell apoptosis. The retinal expression of p38 mitogen-activated protein kinase (p38MAPK), phosphorylated p38MAPK (p-p38MAPK), Bcl-2 and Bax was measured using WB to investigate the mechanism by which hucMSC-Exs affects DRN. Using TEM, NTA and WB, hucMSC-Exs were successfully isolated. No significant change was observed after injection in the normal control group. All rats injected with STZ developed hyperglycemia. HE staining revealed that hucMSC-Exs effectively alleviated retinal structure disruption and reduced the apoptosis of RGCs (P < 0.05). Cells positive for TUNEL (TUNEL+) occurred at a higher rate in the diabetic group than in other groups (P < 0.05). Compared with the normal control group, the expression of p-p38MAPK was significantly increased in the diabetic group and decreased in the hucMSC-Exs group (P < 0.01). The expression of Bax was significantly decreased while Bcl-2 expression was significantly increased in hucMSC-Exs group (P < 0.01). These findings suggest that intravitreal injection of hucMSC-Exs can reduce DRN and protect retinal structure, and that these effects are mediated through inhibition of the p38MAPK pathway.


Recurrent priapism in the young patient treated with baclofen.
Recurrent priapism may lead to erectile dysfunction; its treatment is aimed at the prevention of new episodes. We report a case of idiopathic recurrent priapism in a 15-year-old patient. He was treated with oral baclofen, starting with a daily dose of 10mg that was increased to 30 mg to achieve control of symptoms. After a 6-month follow up, he reported no recurrence or side effects.


Peripheral insulin and amylin levels in Parkinson's disease.
Type-2-diabetes (T2D) has surfaced as a potential risk factor for Parkinson's disease (PD) in some epidemiological studies. Evidence of glucose metabolism alterations in PD from molecular studies remains conflicting. Amylin, the T2D amyloid protein, has been implicated in PD in pathological studies. We aimed to assess peripheral levels of amylin and insulin in PD patients and control subjects (Cs). We conducted an observational cross-sectional study of 111 participants: 73 PD and 38 Cs, similar in age, sex and body mass index. All underwent motor (UPDRS-MDS-III), non-motor (NMSS) and cognitive (MDRS) scales as well as determination of four parameters: fasting glycaemia, glycated haemoglobin, fasting plasma insulin (FPI) and fasting plasma amylin (FPA). FPI was significantly lower in PD than Cs (p = 0.034). In participants with age above cohort-median-age, FPA was higher in PD than Cs (p = 0.046). The FPA/FPI ratio (FPAIR) was significantly higher in PD than Cs (p = 0.024). In PD, modest correlation was found between higher insulin-resistance and NMSS scores. PD patients had lower FPI and increased FPAIR. In older PD subgroup, FPA was increased. The more the insulin resistance, the higher the non-motor scores. These findings provide an additional link between pathophysiology of diabetes and PD. This might be related to a dissociated insulin and amylin secretion in PD, in line with recent evidence of endocrine pancreas role in PD pathogeny.


Treatment of Bullous Pemphigoid in People Aged 80 Years and Older: A Systematic Review of the Literature.
Bullous pemphigoid commonly affects older adults and has a detrimental effect on both quality of life and longevity. Systemic corticosteroids, the mainstay of therapy, may cause significant adverse effects, especially in older patients. Therefore, safer therapeutic options are being sought. The objective of this article was to systematically review the published evidence on the efficacy and safety of different treatment modalities for bullous pemphigoid in older patients. We performed a systematic review of all publications until May 2020 in PubMed, Google Scholar, and the ongoing trials registry of the US National Institutes of Health databases evaluating the efficacy and safety of bullous pemphigoid treatments in patients aged older than 80 years. The primary outcome was complete response. The secondary outcomes were partial response, complete remission on minimal therapy or during tapering, recurrence, adverse events, and mortality. Twenty-eight publications were included: 2 randomized controlled trials, 5 prospective cohort studies, 10 retrospective cohort studies, and 11 case series, with a total of 153 older patients. The overall complete response rate was 31%. Topical corticosteroids had the highest complete response rate (55%) with a low side-effect profile. Biologics (omalizumab and rituximab) were effective in achieving complete remission on minimal therapy (29%) without recurrence, although rituximab was associated with a relatively high mortality rate (29%). Current data suggest that topical corticosteroids are effective and safe and should remain the first line of treatment for bullous pemphigoid in older adults. However, their application is difficult and requires a high-functioning patient, third-party assistance, or a relatively mild disease. Biological agents are effective but warrant meticulous patient selection owing to the relatively high mortality rate associated with rituximab. PROSPERO registration number CRD42020186686.


Does stress induce (para)sex? Implications for Candida albicans evolution.
Theory predicts that stress is a key factor in explaining the evolutionary role of sex in facultatively sexual organisms, including microorganisms. Organisms capable of reproducing both sexually and asexually are expected to mate more frequently when stressed, and such stress-induced mating is predicted to facilitate adaptation. Here, we propose that stress has an analogous effect on the parasexual cycle in Candida albicans, which involves alternation of generations between diploid and tetraploid cells. The parasexual cycle can generate high levels of diversity, including aneuploidy, yet it apparently occurs only rarely in nature. We review the evidence that stress facilitates four major steps in the parasexual cycle and suggest that parasex occurs much more frequently under stress conditions. This may explain both the evolutionary significance of parasex and its apparent rarity.


A yeast replication origin consists of multiple copies of a small conserved sequence.
Autonomously replicating sequences (ARSs) of the yeast S. cerevisiae function as replication origins on plasmids and probably also on chromosomes. ARS function requires a copy of the ARS core consensus (5'-[A/T]TTTAT[A/G]TTT[A/T]-3') and additional sequences 3' to the T-rich strand of the consensus. Our analysis of an ARS from chromosome III, the C2G1 ARS, suggests that ARS function depends on the presence of an exact match to the core consensus and the presence of additional near matches in the 3' flanking region. We have demonstrated that ARS function can be mediated by multiple matches to the core consensus by constructing synthetic ARS elements from oligonucleotides containing copies of the consensus sequence. We find that two copies of the core consensus are sufficient for ARS activity and that an artificial ARS as efficient as a natural chromosomal ARS can be constructed from multiple core consensus elements in a specific orientation.


Daytime melatonin and temazepam in young adult humans: equivalent effects on sleep latency and body temperatures.
1. As changes in core body temperature are generally associated with concomitant changes in sleep propensity, it is possible that the effects of hypnotic/soporific agents may be related to changes in thermoregulation. Therefore, to increase our knowledge of the mechanisms by which these agents exert their soporific effects, we compared the thermoregulatory and soporific effects of temazepam (20 mg per os (p.o.)) with those of melatonin (5 mg p.o.) when administered at 14.00 h to 20 young healthy adults (13 male, 7 female; age, 23.5 +/- 0.4 years). 2. From 08.00 to 20.30 h, subjects lay in bed, and foot and rectal (Tc) temperatures were recorded. Sleep onset latency (SOL) was measured using 20 min multiple sleep latency tests, performed hourly from 11.00 to 20.00 h, during which time heart rate was recorded. 3. Compared with placebo, both melatonin and temazepam significantly reduced Tc (-0.17 +/- 0.02 and -0.15 +/- 0.03 C, respectively) and SOL (by 4.8 +/- 1.49 and 6.5 +/- 1.62 min, respectively). Although both treatments significantly increased heat loss, only melatonin demonstrated cardiac effects. Importantly, there was a temporal relationship between minimum SOL and the maximum rate of decline in Tc for both melatonin (r = 0.48) and temazepam (r = 0.44). 4. A possible role of thermoregulation in sleep initiation is suggested by the similar temporal relationship between Tc and SOL for two different classes of soporific agents.


Natural history and patterns of treatment change in Parkinson's disease: A retrospective chart review.
Parkinson's disease (PD) management seeks to balance the benefits and harms of current medications and evolves as the disease progresses. The natural history of PD and associated patterns of treatment change were analyzed to identify unmet needs in treatment of PD symptoms. Medical charts of patients from clinics across the US diagnosed on or before June 30th, 2014 were retrospectively reviewed. Index date was the first clinic visit, and the post-index period was through study end (June 30th, 2019). Outcomes included the frequency of therapy changes in the post-index period, reasons for therapy change, and adverse events (AE). Patients (n = 203) at index were receiving levodopa-peripheral dopa decarboxylase inhibitor (PDDI) monotherapy (47%), dopaminergic agonist (DA) monotherapy (15%), monoamine oxidase B inhibitor (MAOBI) monotherapy (14%), or combination therapies. The percentage of patients in Hoehn-Yahr disease Stage 1-2 was 52% at index and 20% by the end of the study. Frequencies of motor, non-motor, and neuropsychiatric symptoms increased during the enrollment. Levodopa-PDDI monotherapy and levodopa-PDDI + MAOBI had the lowest rates of therapy changes. Symptom relapse was the most common reason for dose escalation, add-on, and dose reduction, whereas AEs were the most common reason for discontinuation and switching. Dose escalation, add-on, and forward switch were most likely to occur in the first 6 months of treatment. Therapy changes during the study period reflected the challenging and evolving management of PD as the disease progresses. New or add-on symptomatic treatments are needed that are well-tolerated and able to control PD symptoms.


Rapid structural determination of modified pregnane glycosides from Cynanchum forrestii by liquid chromatography-diode-array detection/electrospray ionization multi-stage tandem mass spectrometry.
The fragmentation behaviors of the two types of modified pregnane glycosides from Cynanchum forrestii were investigated by positive ion electrospray ionization multi-stage tandem mass spectrometry equipped with an ion trap analyzer. The spectral data further illuminated the predominance of ESI-MS(n) technique on the identification of pregnane glycosides, especially of two sorts of modified pregnane glycosides with aglycone skeletons of 13,14:14,15-disecopregnane-type and 14,15-secopregnane-type, which differed in the presence of the characteristic [M-46+Na]+ ion. For sugar residues, the fragment ions were analyzed and some possible fragmentation pathways were proposed, especially for 3-demethyl-2-deoxythevetose, the glycosidic cleavage reaction was easier to occur than those of other sugar units in its moiety. The natures and differences of the pregnane cores, and the types and linked sequences of sugar residues were illustrated. According to these conclusions, eight new pregnane glycosides in the fraction of Cynanchum forrestii were structurally elucidated by HPLC/HRMS and HPLC-DAD/ESI-MS(n) techniques. Results of the present studies can benefit the rapid identification and structural determination of analogous constituents in crude plant extracts.


Screening of toll-like receptor signaling pathway-related genes and the response of recombinant glutathione S-transferase A3 protein to thiram induced apoptosis in chicken erythrocytes.
The expression of genes related to the Toll-like receptors (TLRs) signaling pathway were determined. Group A, B and C fed with basal diet and group D, E and F induced TD by feeding a basal diet containing 100 mg·kg-1 thiram. rGSTA3 protein was injected at 20 μg·kg-1 in group B, E and at 50 μg·kg-1 in C, F. Results suggested that lameness and death of chondrocytes were significant on day 14. TLRs signaling pathway related genes were screened based on the transcriptome enrichment, and validated on qPCR. IL-7, TLR2, 3, 4, 5, 7, 15, MyD88, MHC-II, MDA5 and TRAF6 were significantly (p < 0.05) expressed in group E and F as compared to group D on day 14 and 23. IL-7, MHCII, TRAF6, TLR3, TLR5, TLR7, and TLR15 determined insignificant in group D compared to group A on day 23. TD occur in an early phase and alleviated in the later period. rGSTA3 protein can prevent apoptosis and repair degraded chondrocytes.


Criteria for Carotid Atherosclerotic Plaque Instability.
The study aim is to determine the criteria for carotid atherosclerotic plaque instability with the use of an advanced ultrasound technology, immunohistochemical analysis, and electron paramagnetic resonance (EPR) and assess their correlations with histologic results. A total of 92 patients were included in the study and were examined by ultrasound duplex scanning and ultrasound elastography. Plaques harvested during carotid endarterectomy were obtained for histologic analysis, immunofluorescent assay, and EPR spectroscopic measurements. Multivariate logistic regression analysis showed that plaques with an area >90 mm2 (odds ratio [OR], 4.05; 95% confidence interval [CI], 1.32-13.2; P = 0.006), plaque volume index > 0.6 cm3 (OR, 2.72; 95% CI, 1.05-9.58; P = 0.04), and juxtaluminal black area ≥8 mm2 (OR, 2.82; 95% CI, 1.22-6. 23; P = 0.02) were statistically significant independent predictors of histologically verified unstable plaques. Unstable plaques occurred in 94% of the patients with these indicators. Significant increases in the number of CD68+ and CD36+ cells (inflammatory markers) and CD31+ cells (neovasculogenesis markers) were revealed in unstable plaques by the immunohistochemical assay. EPR data analysis showed that divalent manganese could serve as a marker of plaque instability. Additional ultrasound criteria, verified by histologic studies, significantly increased the information content for identifying patients with unstable plaques, which can be of great importance in stratifying the risk of ischemic stroke, especially in asymptomatic patients. The degree of calcification is not a mandatory criterion for plaque stabilization.


Extreme androgen resistance in a kindred with a novel insertion/deletion mutation in exon 5 of the androgen receptor gene.
Androgen insensitivy syndrome (AIS) is the most frequent cause of male pseudohermaphroditism resulting from target-organ resistance to androgen action. Individuals bearing the complete form of the disease (CAIS) present a female phenotype and a lack of pubic and axillary hair. In the present study, four 46,XY patients born in two generations from a kindred with a history of AIS were examined for genetic abnormalities in the androgen receptor gene (AR). All eight exons encoding the AR protein were individually amplified from genomic DNA followed by a mutation screening with single-strand conformation polymorphism analysis. Sequencing of the mutant AR revealed a novel insertion/deletion mutation in exon 5. A deletion of 7 bp is replaced by an insertion of 11 nucleotides, which represents a duplication of the adjacent downstream sequence. The mutation g.2640_2646delAGGATGC/2652_2662insTTCGCCCCTGA, results in a frameshift that introduces a premature termination signal TGA, nine codons downstream. Such a rearrangement predicts a truncation of the AR, thereby deleting a large portion of the ligand-binding domain (amino acid position 768-919). Furthermore, although this mutation breaks the translational reading frame starting from codon 760, examination of the complementary DNA suggested that it does not disturb mRNA splicing. These changes have been found in all the patients and appear to account for the observed absence of detectable androgen binding to the AR in cultured fibroblasts and for the CAIS phenotype in the kindred. This disorder represents the first insertion/deletion mutation of the AR that probably arose by a slipped-strand mispairing mechanism.


Human papillomavirus-driven repression of NRF2 signalling confers chemo-radio sensitivity and predicts prognosis in head and neck squamous cell carcinoma.
To investigate the role of NRF2 signalling in conferring superior prognosis in patients with HPV positive (HPV+ve) head & neck squamous cell carcinomas (HNSCC) compared to HPV negative (HPV-ve) HNSCC and develop molecular markers for selection of HPV+ve HNSCC patients for treatment de-escalation trials. NRF2 activity (NRF2, KEAP1, and NRF2-transcriptional targets), p16, and p53 levels between HPV+ve HNSCC and HPV-ve HNSCC in prospective and retrospective tumor samples as well as from TCGA database were compared. Cancer cells were transfected with HPV-E6/E7 plasmid to elucidate if HPV infection represses NRF2 activity and sensitizes to chemo-radiotherapy. Prospective analysis revealed a marked reduction in expression of NRF2, and its downstream genes in HPV+ve tumors compared to HPV-ve tumors. A retrospective analysis by IHC revealed significantly lower NQO1 in p16high tumors compared to p16low tumors and the NQO1 expression correlated negatively with p16 and positively with p53. Analysis of the TCGA database confirmed low constitutive NRF2 activity in HPV+ve HNSCC compared to HPV-ve HNSCC and revealed that HPV+ve HNSCC patients with 'low NQO1' expression showed better overall survival compared to HPV+ve HNSCC patients with 'high NQO1' expression. Ectopic expression of HPV-E6/E7 plasmid in various cancer cells repressed constitutive NRF2 activity, reduced total GSH, increased ROS levels, and sensitized the cancer cells to cisplatin and ionizing radiation. Low constitutive NRF2 activity contributes to better prognosis of HPV+ve HNSCC patients. Co-expression of p16high, NQO1low, and p53low could serve as a predictive biomarker for the selection of HPV + ve HNSCC patients for de-escalation trials.


Higher MRI lesion load in multiple sclerosis is related to the N-glycosylation changes of cerebrospinal fluid immunoglobulin G.
Intrathecal clonal expansion of antibody-producing plasma cells in multiple sclerosis (MS) perpetuates central nervous system injury and is associated with active demyelination. Immunoglobulin G (IgG) effector functions are modulated by linked N-glycan structures. The aim of the study was to detect potential differences in N-glycosylation of IgG in serum and cerebrospinal fluid (CSF) and total sera proteins between people with MS and those in whom the diagnosis of MS was excluded. Furthermore, we investigated the association with standard laboratory biomarkers of intrathecal inflammation as well as clinical and neuroradiological disease activity. This cross-sectional study included patients with suspected demyelinating disease. MS diagnosis was based on the 2017 McDonald criteria and controls were patients with excluded MS diagnosis. N-glycans were compared with Expanded Disability Status Scale (EDSS), magnetic resonance imaging (MRI) markers of disease activity and biomarkers of intrathecal inflammation (cell count, CSF-IgG concentration, percentage of intrathecal IgG, oligoclonal bands (OCB), virus-specific antibody index (MRZH reaction)). Differences between groups were observed only in the CSF-IgG N-glycome. In MS, the presence of bisecting N-acetylglucosamine (Padj=2.63E-05) and monogalactosylation (Padj=1.49E-06) were more abundant and associated with positive OCBs. N-glycans monogalactosylated at the α6 arm FA2[6]G1 (r = 0.56) and FA2[6]BG1 (r = 0.45) correlated with percentage of intrathecal IgG, but not total CSF-IgG. This trait was also more abundant in MRZH positive people with MS who had higher MRI lesion load (P = 0.018) but unrelated to active lesions or EDSS. More abundant monogalactosylation of intrathecally synthesized IgG is the most prominent trait in MS and is associated with higher MRI lesion load.


A local subset of mesenchymal cells expressing the transcription factor Osr1 orchestrates lymph node initiation.
During development, lymph node (LN) initiation is coordinated by lymphoid tissue organizer (LTo) cells that attract lymphoid tissue inducer (LTi) cells at strategic positions within the embryo. The identity and function of LTo cells during the initial attraction of LTi cells remain poorly understood. Using lineage tracing, we demonstrated that a subset of Osr1-expressing cells was mesenchymal LTo progenitors. By investigating the heterogeneity of Osr1+ cells, we uncovered distinct mesenchymal LTo signatures at diverse anatomical locations, identifying a common progenitor of mesenchymal LTos and LN-associated adipose tissue. Osr1 was essential for LN initiation, driving the commitment of mesenchymal LTo cells independent of neural retinoic acid, and for LN-associated lymphatic vasculature assembly. The combined action of chemokines CXCL13 and CCL21 was required for LN initiation. Our results redefine the role and identity of mesenchymal organizer cells and unify current views by proposing a model of cooperative cell function in LN initiation.


Increased polyamine formation in rat salivary glands by stimulation of their autonomic nerve supply.
Ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis, was measured in the three major salivary glands of rats and mice. Enzyme activity and glandular putrescine content increased after continuous stimulation of the parasympathetic or sympathetic salivary nerves for 3 h. The activity was markedly reduced in animals fasted for 24 h to decrease nervous-reflex activation of the glands. Thus ODC activity may be linked with synthesis of secretory macromolecules.


Expression variations of DNA damage response genes ATM and ATR in blood cancer patients.
Hematological malignancies (HM) constitute a variety of cancers originating in blood, bone marrow (BM), and lymphatic systems. During the last two decades, the incidence of HM has dramatically increased worldwide. The etiology of HM is still debatable. Genetic instability is a major risk factor for HM. DDR network is a complex signal transduction cellular machinery that detects DNA damage and activates cellular repair factors, thus maintaining genomic integrity. DDR network detects a variety of DNA damage and triggers the activation of cell cycle control, DNA repair, senescence, and apoptosis. Among the DNA repairing pathways, the DNA damage response (DDR) pathway includes DNA damage signaling apparatus such as ATM and ATR genes. ATM tends to detect double-strand breaks (DSBs) while ATR detects single-strand DNA (ssDNA). The study was conducted to observe the expression deregulations of DNA damage response (DDR) pathway genes (ATM, ATR) at mRNA level in 200 blood cancer patients and 200 controls. The real-time PCR was used to analyze the expression of the target genes. The expression results showed statistically significant downregulation of ATM (p < 0.0001) and ATR (p < 0.0001) genes in blood cancer patients vs. controls. Moreover, a significant downregulation of ATM (p < 0.0001) and ATR (p < 0.0001) was obtained in chemotherapy-treated patients vs. healthy controls. The results suggest that dysregulation in ATM and ATR genes may be associated with increased blood cancer risk.