Spasticity management with botulinum toxin: development and evaluation of a tool for audit.
To develop and evaluate tools for the audit of spasticity management with botulinum toxin. Audit tools to assess the organisation of services and process of care were developed with a consensus process. The tools were piloted across 8 services using a retrospective case note audit. Inter-rater reliability was assessed, using percentage agreement and kappa scores. Clinicians involved in the pilot were surveyed and qualitative feedback was analysed. Eight services (100%) completed service Organisation tools and 7 (88%) returned process of Care tools. One hundred sets of clinical records were audited, with 34 used to assess inter-rater reliability. Eleven items on the process of care tool demonstrated a good degree of inter-rater agreement, but 6 require further development. In the qualitative analysis clinicians stated that the tools captured indicators of quality, and that they would use them again. They recommended that patient satisfaction was included as a measure of quality. The audit has been used practically in the pilot services to provide an impetus for quality improvement. The majority of the audit questions showed a good level of reliability, and clinician feedback supports face validity but a larger scale evaluation is required.


The Role of Screen Time and Screen Activity in the Nonverbal Reasoning of 5-Year-Olds: Cross-Sectional Findings from a Large Birth Cohort Study.
Family and home environment factors have been outlined in previous literature as important variables that affect early reasoning development. However, little research has focused on the association between screen use in the home environment and nonverbal reasoning ability. The aim of this cross-sectional study is to examine the role of both screen time and various screen activities (e.g., television, video, or educational games) in nonverbal reasoning ability in 9,001 5-year-old children using a large birth cohort study (Growing Up in Ireland). Interviews conducted with parents related to the children's screen use and various family factors, while reasoning ability was measured using a standardized task (Picture Similarities Task, British Ability Scales II). A hierarchical multiple regression examined the role of screen use in nonverbal reasoning, while also statistically controlling for family factors such as parental education and employment status. Screen use variables made a significant contribution to the regression model, even after family factors were accounted for, although the effect sizes were very small. Playing educational games, video games, or engaging in over three hours screen use per day were all significant predictors of nonverbal reasoning scores in the final adjusted model. The results of this study suggest that screen use may play a small role in the development of nonverbal reasoning in young children. The findings highlight the need for further studies in this area and may have implications for current debates in screen time research.


Estimating construction waste generation in the Greater Bay Area, China using machine learning.
Reliable construction waste generation data is a prerequisite for any evidence-based waste management effort, but such data remains scarce in many developing economies owing to their rudimentary recording systems. By referring to several models proposed for estimating waste generation, this study aims to develop a reliable and accessible method for estimating construction waste generation based on limited publicly available data. The study has two objectives. Firstly, it aims to estimate construction waste generation by focusing on the Greater Bay Area (GBA) in China, one of the world's most thriving regions in terms of construction activities. Secondly, it aims to compare the strengths and weaknesses of various waste quantification models. 43 sets of annual socio-economic, construction-related and C&D waste generation data ranging from 2005 to 2019 were collected from the local government authorities. By analyzing the data using four types of machine learning models, namely multiple linear regression, decision tree, grey models, and artificial neural network, it is found that all calibrated models, with their respective strengths and weaknesses, can produce acceptable results with the testing R2 ranging from 0.756 to 0.977. This study also reveals that the 11 cities in the GBA produced a total of about 364 million m3 of construction waste in 2018. The result can be used for monitoring the urban metabolism, quantifying carbon emission, developing a circular economy, valorizing recycled materials, and strategic planning of waste management facilities in the GBA. The research findings also contribute to the methodologies for estimating waste generation using limited data.


Oridonin induces apoptosis in gastric cancer through Apaf-1, cytochrome c and caspase-3 signaling pathway.
To investigate the effect and mechanism of oridonin on the gastric cancer cell line HGC-27 in vitro. The inhibitory effect of oridonin on HGC-27 cells was detected using the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. After treatment with 10 μg/mL oridonin for 24 h and 48 h, the cells were stained with acridine orange/ethidium bromide. The morphologic changes were observed under an inverted fluorescence microscope. DNA fragmentation (a hallmark of apoptosis) and lactate dehydrogenase activity were examined using DNA ladder assay and lactate dehydrogenase-release assay. After treated with oridonin (0, 1.25, 2.5, 5 and 10 μg/mL), HGC-27 cells were collected for anexin V-phycoerythrin and 7-amino-actinomycin D double staining and tested by flow cytometric analysis, and oridonin- induced apoptosis in HGC-27 cells was detected. After treatment with oridonin for 24 h, the effects of oridonin on expression of Apaf-1, Bcl-2, Bax, caspase-3 and cytochrome c were also analyzed using reverse-transcript polymerase chain reaction (RT-PCR) and Western blotting. Oridonin significantly inhibited the proliferation of HGC-27 cells in a dose- and time-dependent manner. The inhibition rates of HGC-27 treated with four different concentrations of oridonin for 24 h (1.25, 2.5, 5 and 10 μg/mL) were 1.78% ± 0.36%, 4.96% ± 1.59%, 10.35% ± 2.76% and 41.6% ± 4.29%, respectively, which showed a significant difference (P < 0.05). The inhibition rates of HGC-27 treated with oridonin at the four concentrations for 48 h were 14.77% ± 4.21%, 21.57% ± 3.75%, 30.31% ± 4.91% and 61.19% ± 5.81%, with a significant difference (P < 0.05). The inhibition rates of HGC-27 treated with oridonin for 72 h at the four concentrations were 25.77% ± 4.85%, 31.86% ± 3.86%, 48.30% ± 4.16% and 81.80% ± 6.72%, with a significant difference (P < 0.05). Cells treated with oridonin showed typical apoptotic features with acridine orange/ethidium bromide staining. After treatment with oridonin, the cells became round, shrank, and developed small buds around the nuclear membrane while forming apoptotic bodies. Lactate dehydrogenase (LDH) release assay showed that after treated with 1.25 μg/mL and 20 μg/mL oridonin for 24 h, LDH release of HGC-27 caused by apoptosis increased from 22.94% ± 3.8% to 52.68% ± 2.4% (P < 0.001). However, the change in the release of LDH caused by necrosis was insignificant, suggesting that the major cause of oridonin-induced HGC-27 cell death was apoptosis. Flow cytometric analysis also revealed that oridonin induced significant apoptosis compared with the controls (P < 0.05). And the apoptosis rates of HGC-27 induced by the four different concentrations of oridonin were 5.3% ± 1.02%, 12.8% ± 2.53%, 28.5% ± 4.23% and 49.6% ± 3.76%, which were in a dose-dependent manner (P < 0.05). After treatment for 24 h, DNA ladder showed that oridonin induced a significant increase in DNA fragmentation in a dose-dependent manner. RT-PCR revealed that mRNA expression levels were up-regulated compared with the controls in caspase-3 (0.917 ± 0.103 vs 0.357 ± 0.019, P < 0.05), cytochrome c (1.429 ± 0.111 vs 1.002 ± 0.014, P < 0.05), Apaf-1 (0.688 ± 0.101 vs 0.242 ± 0.037, P < 0.05) and Bax (0.856 ± 0.101 vs 0.278 ± 0.027, P < 0.05) (P < 0.05), whereas down-regulated in Bcl-2 (0.085 ± 0.012 vs 0.175 ± 0.030, P < 0.05). Western blotting analysis also confirmed this result. Apoptosis of HGC-27 induced by oridonin may be associated with differential expression of Apaf-1, caspase-3 and cytochrome c, which are highly dependent upon the mitochondrial pathway.


Effectiveness of Health Promotion Curriculum in Enhancing Non-communicable Diseases Health Literacy in Indian Youth.
Background There is a theory-praxis gap related to health literacy interventions focused on non-communicable diseases (NCDs) among young people. We designed an NCD curriculum and investigated its' effect on health literacy in non-medical, non-nursing college students in India. We deliberately selected non-medical and non-nursing college students (age 17 to 22 years) as we hypothesized, they would have a minimum baseline knowledge of NCDs. Methods We initially carried out a pilot study on 85 students in a four-day-long workshop (32 teaching hours) using empirically developed health literacy instrument. We administered the curriculum to 120 randomly selected students in four colleges, while 50 students were assigned to the comparison group. The curriculum was given over four days for a total of 32 hours. Approval was sought to give four credits for completion of the course. Each lecture comprised didactics, followed by discussion, and skills testing of measuring blood pressure and blood sugar. Health literacy was measured using a specifically designed tool at baseline and endline. The difference in health literacy scores between the two time-points (timepoint 1: before delivering the curriculum, time-point 2: at the end of four days of training) was analyzed using the t-test. Multiple linear and Poisson regression models were used to account for covariates. Results The average difference between the intervention and the control group in baseline scores was 0.05% points (unpaired t-test statistics: -1.36, degrees of freedom 103.15, p>0.05). The same at endline was 20.59% points (unpaired t-test statistics: -11.31, degrees of freedom 138.14, p<0.001). The endline to baseline difference in health literacy scores was 18.54% points higher in the intervention group versus the control group (unpaired t-test statistics: -10.88, degrees of freedom 161.32, p<0.001). The difference-in-difference scores remained significant after accounting for college setting and baseline score (Multivariable linear regression model, β: 19.62% points, p<0.001). None of the socio-economic characteristics were significantly associated with the difference in the difference scores, independent of the intervention effect. The proportion of participants scoring 40% or above on the health literacy measure at endline was significantly higher in the intervention versus the control group (p<0.001). Conclusions We provide empirical data to support the incorporation of NCDs as a credit course in college curricula in low- and middle-income countries. Our findings showed that a theory-driven skills-focused curriculum may be a tool for enhancing NCD health literacy in Indian youth from diverse academic and socio-economic backgrounds.


The Differences of Quantitative Flow Ratio in Coronary Artery Stenosis with or without Atrial Fibrillation.
Quantitative flow ratio (QFR) is a new method for the assessment of the extent of coronary artery stenosis. But it may be obscured by the cardiac remodeling and abnormal blood flow of the coronary artery when encountering atrial fibrillation (AF). The present study aimed to examine the impact of these changed structures and blood flow of coronary arteries on QFR results in AF patients. Methods and Results. We evaluated QFR in 223 patients (112 patients with AF; 111 non-AF patients served as controls) who had undergone percutaneous coronary intervention (PCI) due to severe stenoses in coronary arteries. QFR of the target coronary was determined according to the flow rate of the contrast agent. Results showed that AF patients had significantly higher QFR values than control (0.792 ± 0.118 vs. 0.685 ± 0.167, p < 0.001). We further analyzed local QFR around the stenoses (0.858 ± 0.304 vs. 0.756 ± 0.146, p=0.002), residual QFR (0.958 ± 0.055 vs. 0.929 ± 0.093, p=0.005), and index QFR (0.807 ± 0.108 vs. 0.713 ± 0.152, p < 0.001) in these two groups of patients with and without AF. Further analysis revealed that QFR in AF patients was negatively correlated with coronary flow velocity (R = -0.22, p=0.02) and area of stenosis (R = -0.70, p < 0.001) but positively correlated with the minimum lumen area (MLA) (R = 0.47, p < 0.001). Conclusion. AF patients with coronary artery stenosis have higher QFR values, which are associated with decreased blood flow velocity, smaller stenosis, and larger MLA in AF patients upon cardiac remodeling.


Jaundice, the circulation and the kidney.
(Fig. 1) Cholemia per se (i.e. independent of parenchymal liver damage) causes a profound disturbance of systemic hemodynamics. This includes decrease in total peripheral vascular resistance, and possible impairment of left ventricular performance. These, in turn, lead to a decrease in effective blood volume, a tendency to hemorrhagic shock and prerenal failure. Early in the course of cholemia , the natriuretic effects of bile salts in the circulation may aggravate the hypovolemia. In marked contrast to the decrease in total peripheral vascular resistance, the regional vascular beds of the kidney and the brain constrict during cholemia . The combined effect of cholemia may thus lead to redistribution of cardiac output away from the kidney and the brain. When parenchymal liver damage complicates obstructive jaundice, the tendency to arterial hypotension is aggravated. The overall interrelationship between jaundice and circulatory homeostasis is depicted in figure 1.


Monoclonal anti-erythrocyte auto-antibodies from unimmunized NZB mice.
In order to study the heterogeneity of the anti-erythrocyte auto-antibody response in NZB mice, we have developed hybridomas producing monoclonal auto-antibodies with anti-erythrocyte activity. These monoclonals were prepared by fusion of NZB splenocytes with the P3 X 63.Ag8 myeloma and were screened for activity by indirect immunofluorescence using flow cytometry. We have produced a variety of monoclonal anti-red cell auto-antibodies that have differing antigenic reactivities and immunoglobulin isotypes. Eleven monoclonal antibodies extensively studied thus far react with intact mouse erythrocytes, whereas only two of the eleven also cross react with sheep erythrocytes and bromelain treated mouse erythrocytes. These results suggest that the fusion pattern may represent the range of anti-red cell auto-antibodies found in the intact NZB mouse, with the exception of monoclonal auto-antibodies against cryptic red cell auto-antigens, which were not demonstrated in this initial fusion study.


Advanced MRI Approaches for Evaluating Common Lower Extremity Injuries in Basketball Players: Current and Emerging Techniques.
Magnetic resonance imaging (MRI) can provide accurate and non-invasive diagnoses of lower extremity injuries in athletes. Sport-related injuries commonly occur in and around the knee and can affect the articular cartilage, patellar tendon, hamstring muscles, and bone. Sports medicine physicians utilize MRI to evaluate and diagnose injury, track recovery, estimate return to sport timelines, and assess the risk of recurrent injury. This article reviews the current literature and describes novel developments of quantitative MRI tools that can further advance our understanding of sports injury diagnosis, prevention, and treatment while minimizing injury risk and rehabilitation time. Innovative approaches for enhancing the early diagnosis and treatment of musculoskeletal injuries in basketball players span a spectrum of techniques. These encompass the utilization of T2 , T1ρ , and T2 * quantitative MRI, along with dGEMRIC and Na-MRI to assess articular cartilage injuries, 3D-Ultrashort echo time MRI for patellar tendon injuries, diffusion tensor imaging for acute myotendinous injuries, and sagittal short tau inversion recovery and axial long-axis T1 -weighted, and 3D Cube sequences for bone stress imaging. Future studies should further refine and validate these MR-based quantitative techniques while exploring the lifelong cumulative impact of basketball on players' knees. LEVEL OF EVIDENCE: 5 TECHNICAL EFFICACY: Stage 2.


Treatment of chronic hepatitis B with human lymphoblastoid interferon: results of a controlled trial.
The aim of this study was to evaluate the efficacy of human lymphoblastoid interferon (Wellferon) in the treatment of chronic hepatitis B virus (HBV) infection. To date 70 patients have entered the study, 33 randomized to receive interferon at doses of 5 MU/m2 i.m. daily for the first 4 weeks and three times weekly for 5 months thereafter and 31 acted as controls. Seventy-nine per cent of the treated group permanently cleared HBV DNA compared with 48% of the control group (p = 0.01): 69% of the treated patients and 38% of the controls who eliminated HBV DNA cleared HBeAg, HBeAb (p = 0.02). Twenty-four per cent of the treated patients and 3% (p = 0.01) of the control group had clearance of HBsAg and seroconversion to HBsAb. From these results it was concluded that interferon had a therapeutic effect on Italian heterosexual patients with chronic hepatitis B viral replication.


[Examination of cholangiogram of obstructive jaundice using MRI--compared with PTC and ERCP].
PTC and ERCP are most often used in diagnosis of obstructive jaundice. We studied the possibility of clinical diagnosis using MRI in 33 cases of obstructive jaundice. A clinical diagnosis of malignant tumors could be given in 17 cases out of 20 (85%) using MRI if respiratory standstill was possible. An MRI cholangiogram was particularly effective in describing tearful parting bile ducts and was clearer than PTC in describing negative gallbladders. Choledochal stones could be diagnosed in 58% of cases, which was less than the rate for malignant tumors. MRI is not an invasive examination, can be used in diagnosis of obstructive jaundice, and helps in selecting treatment methods such as PTCD and ERBD.


Osteoporosis and spinal fractures in men with prostate cancer: risk factors and effects of androgen deprivation therapy.
We determined the risk factors for osteoporosis and spinal fractures in men with prostate cancer receiving androgen deprivation therapy. We performed a retrospective analysis of 87 consecutive men with prostate cancer receiving androgen deprivation therapy referred for evaluation of osteoporosis. Data were comprised of lateral thoracolumbar radiographs, bone densitometry, serum biochemistry and a detailed assessment of osteoporotic risk factors. Multivariate regression analysis was used to determine the major risk factors for osteoporosis and spinal fractures. There were 38 (44%) men who were 74.5 years old with radiographic evidence of spinal fractures. They had an initial mean prostate specific antigen of 52.8 ng/ml and had received androgen deprivation therapy for a mean of 39.6 months (95% confidence interval 28.7 to 50.4). Mean spinal (quantitative computerized tomography t-score -4.2) and femoral neck bone mineral densities (dual energy x-ray absorptiometry t-score -2.1) were significantly lower than in men without spinal fractures (p < 0.001 for all measurements). In the regression analysis the duration of androgen deprivation therapy (p = 0.002), serum 25-hydroxyvitamin D levels (p = 0.003) and a history of alcohol excess (defined as more than 4 standard drinks daily, p = 0.04) were the main determinants of spinal fractures. Prolonged androgen deprivation therapy, low serum 25-hydroxyvitamin D levels and a history of alcohol excess are important risk factors for osteoporosis and spinal fractures in men with prostate cancer.


Prostaglandin synthetase inhibitors in pregnancy.
The use of indomethacin as a tocolytic agent in patients with premature labor was first reported by Zuckerman in 1974. Since then, prostaglandin synthetase inhibitors (PSI) have been used in pregnancy for a variety of indications. However, serious fetal side effects of this category of drugs continue to be reported. We present a review of the literature and the experience at our institution on the use of indomethacin, the most commonly prescribed antiprostaglandin in pregnancy. Therapeutic guidelines and surveillance recommendations for the use of PSI in pregnancy are proposed. The role of newer PSI in pregnancy and their side effect profile are also presented.


The PE_PGRS Proteins of Mycobacterium tuberculosis Are Ca(2+) Binding Mediators of Host-Pathogen Interaction.
The phenomenal success of Mycobacterium tuberculosis (M.tb) as a pathogen is primarily based on its ability to modulate host immune responses. The genome of M.tb encodes multiple immunomodulatory proteins, including several members of the multigenic PE_PPE family of which the PE_PGRS proteins are a subset. Curiously, 56 of the 61 PE_PGRS proteins contain multiple copies of the glycine-rich sequence motif GGXGXD/NXUX, a nonapeptide sequence predicted to bind Ca(2+), but the functional significance of these motifs remains a mystery. Here we provide evidence via isothermal titration calorimetry, (45)Ca blotting, fluorescence, and circular dichroism spectroscopy that Ca(2+) binds to the PE_PGRS proteins, PE_PGRS33 (Rv1818c) (10 motifs) and PE_PGRS61 (Rv3653) (one motif). Ca(2+) was observed not to bind to PE_PGRS8 (Rv0742), which lacks nonapeptide motifs. Using recombinant Mycobacterium smegmatis strains expressing Rv1818c and Rv3653 and the THP-1 macrophage model of infection, we show that the two proteins mediate Ca(2+)-dependent upregulation of the anti-inflammatory cytokine IL-10, events critical to the pathogenesis of M.tb. Both Rv1818c and Rv3653 interact with TLR2 in a Ca(2+)-dependent manner, providing a novel mechanistic basis for their immunomodulatory effects. Mutations in the nonapeptide motif of Rv3653 led to compromised Ca(2+) binding, validating the functional criticality of this motif. This study demonstrates for the first time not only their Ca(2+) binding properties but also an essential role for Ca(2+) in the functioning of the M.tb PE_PGRS proteins, opening up the possibility of developing novel anti-tuberculosis therapeutics that inhibit Ca(2+)-PE_PGRS binding.


α-Synuclein Enhances Cadmium Uptake and Neurotoxicity via Oxidative Stress and Caspase Activated Cell Death Mechanisms in a Dopaminergic Cell Model of Parkinson's Disease.
This study examined the role of alpha-synuclein in regulating cadmium (Cd)-induced neurotoxicity using the N27 dopaminergic neuronal model of Parkinson's disease (PD) that stably expresses wild-type human α-synuclein (α-Syn) or empty vector (Vec) control. We report that α-Syn significantly increased Cd-induced cytotoxicity as compared to Vec control cells upon 24 h exposure. To explore the cellular mechanisms, we examined oxidative stress, caspase activation, and Cd uptake and intracellular accumulation. Expression of α-Syn coupled with Cd-induced cytotoxicity increased oxidative stress. Inductively coupled plasma-mass spectrometry (ICP-MS) revealed an increase in Cd uptake and intracellular accumulation in α-Syn-expressing cells upon Cd exposure. Analysis of the mitochondrial mediated apoptotic pathway showed greater activation of caspase-9 and caspase-3 in α-Syn cells. To functionally evaluate the role of metal transporters in the altered Cd phenotype, we examined Cd toxicity in the presence of nontoxic levels of divalent manganese Mn(II) and iron Fe(II). Co-treatment with Fe(II) or Mn(II) did not significantly attenuate Cd-induced cytotoxicity. We report that Cd exposure decreased the divalent metal transporter 1 and Akt protein levels in the α-Syn-expressing cells without altering native PKCδ protein levels in both Vec control and α-Syn lines. In addition, we show decreased basal metallothionein-3 protein expression in α-Syn-expressing cells. Co-treatment with N-acetyl-L-cysteine was sufficient to attenuate and abolish the α-Syn × Cd-induced cytotoxicity. Collectively, these results demonstrate that α-Syn exhibits neurotoxic properties upon acute Cd exposure to cause cell death by causing oxidative stress, increasing Cd uptake, altering caspase-9 and caspase-3 activation, and diminishing the neuroprotective effect of Akt in a dopaminergic neuronal model of PD.


Antisera Produced Using an E. coli-Expressed SARS-CoV-2 RBD and Complemented with a Minimal Dose of Mammalian-Cell-Expressed S1 Subunit of the Spike Protein Exhibits Improved Neutralization.
E. coli-expressed proteins could provide a rapid, cost-effective, and safe antigen for subunit vaccines, provided we can produce them in a properly folded form inducing neutralizing antibodies. Here, we use an E. coli-expressed SARS-CoV-2 receptor-binding domain (RBD) of the spike protein as a model to examine whether it yields neutralizing antisera with effects comparable to those generated by the S1 subunit of the spike protein (S1 or S1 subunit, thereafter) expressed in mammalian cells. We immunized 5-week-old Jcl-ICR female mice by injecting RBD (30 µg) and S1 subunit (5 µg) according to four schemes: two injections 8 weeks apart with RBD (RBD/RBD), two injections with S1 (S1/S1), one injection with RBD, and the second one with S1 (RBD/S1), and vice versa (S1/RBD). Ten weeks after the first injection (two weeks after the second injection), all combinations induced a strong immune response with IgG titer > 105 (S1/RBD < S1/S1 < RBD/S1 < RBD/RBD). In addition, the neutralization effect of the antisera ranked as S1/RBD~RBD/S1 (80%) > S1/S1 (56%) > RBD/RBD (42%). These results indicate that two injections with E. coli-expressed RBD, or mammalian-cell-produced spike S1 subunit alone, can provide some protection against SARS-CoV-2, but a mixed injection scheme yields significantly higher protection.


Trajectory of Systemic Blood Pressure in Early Life: A Cohort Study.
Objective. To track the BP (blood pressure) trajectory of healthy infants during the first year of life of healthy infants born in Northeast Brazil. Methods. In this cohort study, BP was assessed by oscillometry at the first 24 hours of life and 12 months of age. Results. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) increased with age (P < .001) and were not influenced by gender (SBP: P = .178 and DBP: P = .623) or type of delivery (SBP: P = .827 and DBP: P = .106), when compared between the first 24 hours of life and 12 months of age. Conclusion. The data from the present study increased knowledge about the trajectory of BP during the first year of life. The increase in BP between the first month and the first year of life was not influenced by gender or type of delivery.


The Role of Radiofrequency Hyperthermia in The Radiosensitization of A Human Prostate Cancer Cell Line.
This study evaluated enhanced induced DNA damages and apoptosis of a spheroid culture of DU145 prostate cancer cells treated by a combination of radiofrequency hyperthermia (RF HT) with radiation treatment (RT) from an external radiotherapy machine compared to RT alone. In this experimental study, DU145 cells were cultured as spheroids until they reached 300 µm in diameter. We exposed these cultures to either: RF HT for 90 minutes at 43˚C originated from a Celsius TCS system, RF HT followed by RT at doses of 2 Gy or 4 Gy (15 MV energy) with 15-minute interval, or RT alone at the above mentioned doses. The trypan blue exclusion assay, alkaline comet assay, and annexin V/PI flow cytometry were performed to measure cell viability, the amount of DNA damage in an individual cell as the tail moment, and percentage of induced cell apoptosis in response to treatments explained. We calculated the thermal enhancement factor (TEF) for the combined treatment regime. RF HT followed by the 4 Gy dose of RT resulted in minimum viability (85.33 ± 1.30%), the highest tail moment (1.98 ± 0.18), and highest percentage of apoptotic cells (64.48 ± 3.40%) compared to the other treatments. The results of the TEF assay were 2.54 from the comet assay and 2.33 according to flow cytometry. The present data suggest that combined treatment of mega voltage X-rays and RF HT can result in significant radiosensitization of prostate cancer cells.


Recurrent osteochondroma of the mandibular condyle: A case report.
A 21-year-old woman presented with facial asymmetry. Crepitus and clicking of the temporomandibular joint were noted. The midline deviated 5.5 mm to the left, and secondary malocclusion was observed. Panoramic and cone-beam computed tomographic images showed an irregular and exophytic bony mass on the anteromedial surface of the right mandibular condyle. A 3-phase bone scan revealed increased tracer uptake on the affected side. The lesion was treated with excision and reshaping under the diagnosis of osteochondroma confirmed by a histopathological examination. The lesion recurred after 3 years, and the patient underwent condylectomy. Mandibular condylar osteochondroma is often resected because it causes functional and aesthetic problems, but it rarely recurs. To the best of our knowledge, only 2 cases of recurrent osteochondromas of the mandibular condyle have been reported previously. Surgical treatment of the osteochondroma should be performed considering the possibility of recurrence, and long-term follow-up is recommended.


The impact of fasting plasma glucose variability on osteoporotic fractures.
To investigate the impact of FPG variability on osteoporotic fractures in the entire community population. All participants were from the Kailuan Study. Participants completed three consecutive surveys from 2006-2007, 2008-2009, and 2010-2011. We excluded individuals with an osteoporotic fracture in or prior to the index year and those without complete FPG records at the first 3 examinations. All participants were followed from the date of the 3rd examination to the first occurrence of an endpoint event or December 31, 2021. According to the SD of FPG levels, the included subjects were divided into three groups. A Cox proportional hazards model was performed to further analyze the effect of different FPG-SD groups on the risk of osteoporotic fractures. Ultimately, the study population included 57295 participants. During a median follow-up time of 11.00 years, we documented 772 new osteoporotic fracture cases. When evaluating the FPG-SD level as a categorical variable, the HRs for osteoporotic fractures were 1.07 (95% CI: 0.89-1.29) for T2 and 1.32 (95% CI: 1.10-1.60) for T3 when compared with T1. We found that increased FPG variability was associated with a greater risk of osteoporotic fractures in people with diabetes than in those without diabetes (47% vs. 32%). Increased FPG variability was an independent predictor of incident osteoporotic fracture, especially in individuals older than 50 years old, nonobese individuals, diabetes patients, and individuals with positive FPG-SD variability.


Different subcellular localizations and functions of human ARD1 variants.
ARD1 is present in various species and has several variants derived from alternative splicing of mRNA. Previously, we reported differential biological functions and cellular distributions of mouse ARD1 (mARD1) variants. However, in comparison to mARD1 variants, human ARD1 (hARD1) variants have been rarely studied. In this study, we characterized a hARD1 variant, hARD1(131) and investigated its cellular activities. hARD1(131) mRNA was isolated from HeLa cells and sequenced. Sequence alignment revealed that, compared to hARD1(235), the most common form of hARD1, the mRNA sequence encoding hARD1131 possesses an altered reading frame due to a 46-bp deletion. Thus, hARD1(131) and hARD1(235) differ in their C-terminal regions with a partially deleted acetyltransferase domain at the C-terminus of hARD1(131). Moreover, hARD1(131) and hARD1(235) showed different subcellular localizations and biological functions. hARD1(131) was mostly localized in the cell nucleus, whereas hARD1(235) was primarily localized in the cytoplasm. In addition, hARD1(235) stimulated cell proliferation by upregulation of cyclin D1, however hARD1(131) had no influence on cyclin D1 expression and cell growth. Because hARD1(235) enhances cell proliferation by its autoacetylation activity, we examined the autoacetylation activity of hARD1(131) and observed that this function was absent in hARD1(131). These results suggest that human ARD1 variants have different effects on cell prolifer-ation, which may result from distinct subcellular localizations and autoacetylation activities.


Lack of amyloidosis and renal disease in A strain mice.
Amyloid deposition in two sublines of the A strain mouse, a reputedly reliable murine model of spontaneous amyloidosis, was reevaluated using combined green polarization birefringence after Congo red staining and a fibrillar ultrastructure for amyloid identification. Both sublines were found to be devoid of amyloid at an age when the A strain mouse had been reported to have a high incidence of papillonephritis, amyloidosis, and death. This finding indicates a need for reassessment of this and other traditional models of spontaneous amyloidosis.


Unsupervised Reconstruction of Analyte-Specific Mass Spectra Based on Time-Domain Morphology with a Modified Cross-Correlation Approach.
Concomitant species that appear at the same or very similar times in a mass-spectral analysis can clutter a spectrum because of the coexistence of many analyte-related ions (e.g., molecular ions, adducts, fragments). One method to extract ions stemming from the same origin is to exploit the chemical information encoded in the time domain, where the individual temporal appearances inside the complex structures of chronograms or chromatograms differ with respect to analytes. By grouping ions with very similar or identical time-domain structures, single-component mass spectra can be reconstructed, which are much easier to interpret and are library-searchable. While many other approaches address similar objectives through the Pearson's correlation coefficient, we explore an alternative method based on a modified cross-correlation algorithm to compute a metric that describes the degree of similarity between features inside any two ion chronograms. Furthermore, an automatic workflow was devised to be capable of categorizing thousands of mass-spectral peaks into different groups within a few seconds. This approach was tested with direct mass-spectrometric analyses as well as with a simple, fast, and poorly resolved LC-MS analysis. Single-component mass spectra were extracted in both cases and were identified based on accurate mass and a mass-spectral library search.


626-nm single-frequency semiconductor laser system operated near room temperature for mW-level second-harmonic generation at 313 nm.
We report a 626-nm, single-frequency generation based on laser diodes operated near room temperature and the second-harmonic generation with an output power of 2 mW at 313 nm. A fundamental single-frequency laser at 626 nm consists of an external-cavity diode laser and a tapered semiconductor amplifier with 200 mW output. Antireflection coating of the laser diode expands the wavelength tuning range and realizes single-frequency output at 626 nm at room temperature. 313 nm light is generated by cavity-enhanced frequency doubling with a β-BaB2O4 crystal. A 313 nm laser with mW-level output power is useful for various experiments using laser cooling of beryllium ions.


Double-Labeling Method for Visualization and Quantification of Membrane-Associated Proteins in Lactococcus lactis.
Lactococcus lactis displaying recombinant proteins on its surface can be used as a potential drug delivery vector in prophylactic medication and therapeutic treatments for many diseases. These applications enable live-cell mucosal and oral administration, providing painless, needle-free solutions and triggering robust immune response at the site of pathogen entry. Immunization requires quantitative control of antigens and, ideally, a complete understanding of the bacterial processing mechanism applied to the target proteins. In this study, we propose a double-labeling method based on a conjugated dye specific for a recombinantly introduced polyhistidine tag (to visualize surface-exposed proteins) and a membrane-permeable dye specific for a tetra-cysteine tag (to visualize cytoplasmic proteins), combined with a method to block the labeling of surface-exposed tetra-cysteine tags, to clearly obtain location-specific signals of the two dyes. This allows simultaneous detection and quantification of targeted proteins on the cell surface and in the cytoplasm. Using this method, we were able to detect full-length peptide chains for the model proteins HtrA and BmpA in L. lactis, which are associated with the cell membrane by two different attachment modes, and thus confirm that membrane-associated proteins in L. lactis are secreted using the Sec-dependent post-translational pathway. We were able to quantitatively follow cytoplasmic protein production and accumulation and subsequent export and surface attachment, which provides a convenient tool for monitoring these processes for cell surface display applications.


Feeding and digestion in low salinity in an osmoconforming crab, Cancer gracilis. II. Gastric evacuation and motility.
Gastric evacuation and gut contraction rates were followed in the graceful crab Cancer gracilis during exposure to low salinity. Crabs were fed a radio-opaque meal and then exposed to 100% seawater (SW), 80%SW or 60%SW; passage of digesta was followed using a fluoroscope. Exposure to low salinity increased the time for food passage through the gut system. Times for emptying of the foregut, midgut and hindgut varied in a dose-dependent manner. In the lowest salinity, crabs regurgitated food from the foregut after approximately 6 h. This may act as a protective response, clearing the gut and avoiding subsequent increases in metabolism associated with digestion. Contraction rate of the cardiac stomach and gastric mill was sporadic and there was no significant change with salinity. In contrast, contractions of the pyloric region were more constant and rapid. Pyloric contractions decreased at each salinity within 2-4 h after feeding. Contraction rates of the pyloric chamber were significantly lower in 60%SW compared with 100%SW and 80%SW. During a salinity cycle there was also slowing of gut contractions and food passage through the gut system. Pre-treatment levels were only regained slowly when the animals were returned to 100%SW. Cancer gracilis was able to slow digestion during low salinity exposure, which may spare resources for other systems. However, the crabs could not halt digestion completely and may be committed to protein synthesis once intracellular digestion has begun.


A Urea Potentiometric Biosensor Based on a Thiophene Copolymer.
A potentiometric enzyme biosensor is a convenient detector for quantification of urea concentrations in industrial processes, or for monitoring patients with diabetes, kidney damage or liver malfunction. In this work, poly(3-hexylthiophene-co-3-thiopheneacetic acid) (P(3HT-co-3TAA)) was chemically synthesized, characterized and spin-coated onto conductive indium tin oxide (ITO) glass electrodes. Urease (Urs) was covalently attached to the smooth surface of this copolymer via carbodiimide coupling. The electrochemical behavior and stability of the modified Urs/P(3HT-co-3TAA)/ITO glass electrode were investigated by cyclic voltammetry, and the bound enzyme activity was confirmed by spectrophotometry. Potentiometric response studies indicated that this electrode could determine the concentration of urea in aqueous solutions, with a quasi-Nernstian response up to about 5 mM. No attempt was made to optimize the response speed; full equilibration occurred after 10 min, but the half-time for response was typically <1 min.


Towards a non-ecumenical interchange: Engelhardt, Hauerwas, and Ramsey on Christian bioethics.
Does a non-ecumenical journal on Christian bioethics make sense? Taking issue with Stanley Hauerwas' critique of Ramsey, the author argues (1) interdenominational exchange should not be construed as contest, and (2) the attempt on the part of Christians to address secular issues in secular terms should not be mistrusted or viewed as a contamination hazard. Instead (1) an awareness of human limits should render adherents of different traditions willing to learn from each other and (2) one should see in the love-of-neighbor principle an obligation to serve the world. Addressing the communication problem, the author recommends different language levels for intra-group and ever more encompassing inter-group exchanges.


Emergence of Magnusiomyces capitatus infections in Western Nepal.
Magnusiomyces capitatus is an emerging opportunistic yeast in the Mediterranean region. We report from Nepal one case of M. capitatus infection and six other cases of colonization/probable infection due to M. capitatus at a tertiary care center. Majority of the patients were immunocompromised, at extreme age, associated with comorbidities, and had history of close contact with livestock and poultry. The isolates were identified by phenotypic and genotypic (ITS and D1/D2 region of 26S rDNA sequence) methods. Molecular typing of the isolates was carried out by amplified fragment length polymorphism. Minimum inhibitory concentration (MIC) of the isolates for amphotericin B, caspofungin, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, and micafungin were 2, 0.1-4, 2, 0.12-0.5, 0.12-0.5, 0.25, 1-4, and 1-4 μg/ml, respectively. Presence of M. capitatus infection was not known in Nepal, and the study should alert the clinicians and infectious disease specialists.


Coating noble metal nanocrystals (Ag, Au, Pd, and Pt) on polystyrene spheres via ultrasound irradiation.
The method of ultrasound irradiation is used for anchoring metallic nanocrystals (Ag, Au, Pd, and Pt) onto the surface of polystyrene spheres. In former studies, almost all the sonochemically prepared, coated metallic nanomaterials were formed as amorphous nanoparticles (Pol, V. G.; et al. Langmuir 2002, 18, 3352; Pol, V. G.; et al. Chem. Mater. 2003, 15, 1111; Zhong, Z. Y.; et al. Chem. Mater. 1999, 11 (9), 2350; Pol, V. G.; et al. Chem. Mater. 2003, 15, 1378), which were coated on various substrates (silica spheres, carbon spherules, titania, and alumina). On the other hand, the noble metal nanoparticles deposited on polystyrene spheres via ultrasound irradiation yielded nanocrystalline Ag, Au, Pd, and Pt particles on the surface of polystyrene as as-synthesized materials. The sonochemical mechanism is proposed based on chemical interactions between the particles.


[Clinicopathologic correlation in 50 cases of acquired immunodeficiency syndrome. Retrospective study].
To analyze myocardial abnormalities in patients of acquired immunodeficiency syndrome with clinical and pathological correlation. We selected 50 cases, retrospectively, age ranged from 3 months to 40 years, all of them had myocardial changes and the data of clinical records fulfilled our protocol. Cases of others cardiac diseases were not included. The pathological findings were: myocarditis in 33 (11 had severe myocarditis) and degenerative hystological lesions in 17. The etiologic agents detected were: Toxoplasma in 11, Cryptococcus in 7 and Cytomegalovirus in 3. In 12 cases we could not find any agent. In 15 cases occurred others lesions: endocarditis, pericarditis and sarcoma of Kaposi. It was noted tachycardia in 15 cases, decrease of heart sounds in 12, arterial hypotension in seven, systolic murmur in 8, galop rhythm in 7, pericardial friction rub in 3, arrhythmia in 2. Four patients had congestive heart failure. The EKG showed sinus tachycardia in 18, ST and T changes in 10, low voltage in 5, ST segment elevation in 5 and extrasystoles in 3 cases. The echocardiogram findings were: pericardial effusion in 9 cases and 9 had ventricular dysfunction. The cardiac lesions were very important even in patients without clinical signals. We need others prospective studies with viral identification trying to detect specific lesions of HIV.


The use of optokinetic stimulation in rehabilitation of the hemineglect disorder.
The aim of this study was to investigate whether it is possible to strengthen the rehabilitation of spatial hemineglect by combining standard training for spatial scanning with optokinetic stimulation. A simple randomized design was used: one group of neglect patients was treated with a combination of the two techniques, and a second group received only the standard treatment. Both treatments were given for six consecutive weeks and produced significant improvements. However, addition of the optokinetic stimulation did not improve the patients' performance. Also, none of the independent variables (i.e., presence/absence of hemianopia) predicted the positive effect occasionally observed in individual patients.


Effect of Traditional Treatment of Diabetes Mellitus with Xanthosoma sagittifolium on the Male Reproductive System of Alloxan-Induced Diabetic Wistar Rats.
The management of diabetes mellitus (DM) targets glycemic control as well as prevention or reversal of other complications associated with the metabolic disorder. One of such complications is the infertility which has been traced to oxidative stress and DNA damage caused by DM. In this study, experimentally-induced diabetic male Wistar rats were fed with Xanthosoma sagittifolium (cocoyam) for 14 days and its effect on sperm morphology and characteristics, as well as histology of the testes were assessed. X. sagittifolium corm feed (Xs) were reconstituted with commercial rat feed to contain 25, 50, 75 or 100% X. sagittifolium on a weight/weight basis. One primary and 7 secondary cell abnormalities were observed with overall increased total abnormal sperm cells in the diabetic rats. Sperm cell motility was significantly (p<0.05) reduced in all diabetic rats, but diabetic rats fed with Xs showed increased motility and liveability. Sperm volume was unchanged, but the depressed sperm count in the untreated diabetic rats was increased in Xs-fed rats. Histology of Xs-fed diabetic rats showed reduction in seminiferous tubule diameter and luminal diameter which was significantly enlarged in untreated diabetic rats. X. sagittifolium significantly (p<0.05) prevented further deterioration of sperm cell quality and reversed cell apoptosis as presented by increased sperm motility, liveability and count. In the overall, the chances of fertilization by sperm cells from diabetic patients will be increased as well as achievement of good glycemic control and prevention of further damage by the disease condition.


Simultaneous determination of flavonoids in Ixeridium gracile by micellar electrokinetic chromatography.
A micellar electrokinetic chromatography (MEKC) method has been developed for the quantitative analysis of five flavonoids: luteolin 7-O-glucoside (LG), 2',4'-dihydroxy-dihydrochalcone (DD), 2',4'-dihydroxy-chalcone (DC), 7-hydroxy-flavanone (HF) and quercetin 3-O-galactoside (QG) in Ixeridium gracile with UV detection at 275 nm. The applied voltage was 25 kV and the capillary temperature was kept constant at 25 degrees C. The effects of buffer pH, the concentration of electrolyte and organic modifier on migration behavior were studied. Optimum separation condition was achieved with 15 mM borate, 30 mM sodium dodecyl sulfate (SDS) and 10% (v/v) ethanol at pH 10.5. Regression equations showed good linear relationships (correlation coefficients: 0.9984, 0.9991, 0.9994, 0.9995 and 0.9997) between the peak area of each compound and their concentrations. The relative standard deviations (R.S.D.) of the migration time and peak area were less 1.67 and 3.53% (intra-day), and 1.82 and 3.73% (inter-day), respectively, under the optimum separation conditions. The contents of the five active compounds in I. gracile were determined with satisfactory repeatability and recovery.


Spartan/C1orf124, a reader of PCNA ubiquitylation and a regulator of UV-induced DNA damage response.
PCNA is a key component of DNA replication and repair machineries. DNA damage-induced PCNA ubiquitylation serves as a molecular mark to orchestrate postreplication repair. Here, we have identified and characterized Spartan, a protein that specifically recognizes ubiquitylated PCNA and plays an important role in cellular resistance to UV radiation. In vitro, Spartan engages ubiquitylated PCNA via both a PIP box and a UBZ domain. In cells, Spartan is recruited to sites of UV damage in a manner dependent upon the PIP box, the UBZ domain, and PCNA ubiquitylation. Furthermore, Spartan colocalizes and interacts with Rad18, the E3 ubiquitin ligase that modifies PCNA. Surprisingly, while Spartan is recruited by ubiquitylated PCNA, knockdown of Spartan compromised chromatin association of Rad18, monoubiquitylation of PCNA, and localization of Pol η to UV damage. Thus, as a "reader" of ubiquitylated PCNA, Spartan promotes an unexpected feed-forward loop to enhance PCNA ubiquitylation and translesion DNA synthesis.


Dynamic Gradient Directed Molecular Transport and Concentration in Hydrogel Films.
Materials which selectively transport molecules along defined paths offer new opportunities for concentrating, processing and sensing chemical and biological agents. Here, we present the use of traveling ionic waves to drive molecular transport and concentration of hydrophilic molecules entrained within a hydrogel. The traveling ionic wave is triggered by the spatially localized introduction of ions, which through a dissipative ion exchange process, converts quaternary ammonium groups in the hydrogel from hydrophilic to hydrophobic. Through a reaction-diffusion process, the hydrophobic region expands with a sharp transition at the leading edge; it is this sharp gradient in hydrophilicity that drives the transport of hydrophilic molecules dispersed within the film. The traveling wave moved up to 450 μm within 30 min, while the gradient length remained 20 μm over this time. As an example of the potential of molecular concentration using this approach, a 70-fold concentration of a hydrophilic dye was demonstrated.


[Pivotal role of skeletal tissues in the regulation mechanisms for physiological functions mediated by multiple organ networks].
Bone formation and maintenance are sophisticatedly orchestrated through a well-organized and highly regulated mechanism by two distinct cell types; bone-forming osteoblast and bone-resorbing osteoclast. It has been previously established that the adipocyte-derived hormone leptin regulates bone metabolism through the central nervous system and the sympathetic nervous system. We recently identified the osteoblast as the principal cell type in which the sympathetic tone could signal to regulate bone mass by generating and analyzing the cell specific adrenergic receptor deletion mice. The fact that adipocyte-derived hormone regulates bone metabolism implies that the skeleton might exert a feedback control of glucose metabolism. We then revealed that the skeleton acts as an endocrine regulator of energy metabolism through the osteoblast-specific secreted molecule osteocalcin that activates insulin secretion by pancreatic β-cells, insulin sensitivity in fat, liver, and muscle. Moreover, we have recently reported that the sympathetic tone into osteoblast is a pivotal mediator of leptin regulation of insulin secretion by regulating osteocalcin bioactivity. This unexpected functional cross talk between fat, nervous systems, and skeleton illustrates the importance of the skeleton for the regulation of major physiological functions such as glucose homeostasis in vertebrates.


Nuclear localization of signal transducer and activator of transcription 3 in head and neck squamous cell carcinoma is associated with a better prognosis.
A high frequency of head and neck squamous cell cancers (HNSCC) contain constitutively activated signal transducer and activator of transcription 3 (STAT3). To further elucidate the prognostic role of STAT3 in HNSCC, the expression pattern of STAT3 was correlated with outcome in two independent data sets. STAT3 protein expression analysis was done on a test cohort of 102 patients with HNSCC recruited between 1992 and 2005. Automated quantitative analysis was used to assess STAT3 protein expression. We evaluated associations with clinicopathologic parameters and survival prognosis. Associations were validated in a second, independent cohort of 58 patients with confirmed HNSCC enrolled in the Early Detection Research Network-sponsored study who underwent surgical resection with curative intent at the University of Pittsburgh Medical Center between 2000 and 2004. STAT3 displayed mixed nuclear and cytoplasmic staining. Survival analysis showed that high nuclear STAT3 expression (top tertile versus the rest) was associated with longer progression-free survival (n = 70, mean survival of 88.9 versus 46.7 months, P = 0.012 for the first cohort; n = 37, mean survival of 60.3 versus 33.0 months, P = 0.009 for the second cohort). After best model selection in the multivariable analysis context, only STAT3 was significant, revealing a lower risk of progression and death for patients with high nuclear STAT3-expressing tumors (hazard ratio, 0.28; 95% confidence interval, 0.10-0.82; P = 0.019; and hazard ratio, 0.23; 95% confidence interval, 0.07-0.76; P = 0.016, respectively). Our results indicate that high nuclear STAT3 expression levels by automated quantitative analysis are associated with favorable outcome in HNSCC.


Curcumin-induced cell cycle arrest and apoptosis in human acute promyelocytic leukemia HL-60 cells via MMP changes and caspase-3 activation.
Curcumin (diferuloylmethane), is a natural product derived from the root of the plant Curcuma longa. For centuries, it has been used as a spice and as a herbal medicine in Chinese populations. Curcumin has been shown to inhibit cell proliferation, cell cycle arrest, cyclooxygenase (COX)-1 and -2 expression and apoptosis in several human cancer cell lines. The aim of this investigation was to clarify the mechanisms by which curcumin induced cytotoxicity and apoptosis in human leukemia HL-60 cells. The effects of curcumin on the levels of reactive oxygen species (ROS), Ca+2 production, cyclin E, cdc25c, wee1, Bcl-2, Bax, the changes of mitochondrial membrane potential (MMP), cytochrome c release and the activation of caspase-3 were also investigated in the HL-60 cells. Results of flow cytometry and DAPI staining assays indicated that curcumin induced cytotoxicity and apoptosis in the examined cells. The results from flow cytometry assay indicated that curcumin induced ROS and Ca+2 productions, decreased the levels of MMP and increased the activity of caspase-3, leading to cell apoptosis. Western blot assay also revealed that curcumin increased the levels of Bax and the release of cytochrome c, and decreased the levels of Bcl-2 in the examined cells. The inhibition of caspase-3 activation by z-VAD-fmk (broad-spectrum caspase inhibitor) completely blocked curcumin-induced apoptosis in HL-60 cells.


Zebrafish sod1 and sp1 expression are modulated by the copper ATPase gene atp7a in response to intracellular copper status.
Copper is an essential trace metal for physiological functions, whereas copper overload causes cytotoxicity in living organisms. Genetically determined systems regulate acquisition, distribution and storage for copper maintenance and homeostasis. The Human ATP7A copper transport ATPase modulates intracellular copper distribution, which is critical for copper-dependent enzymes such as superoxide dismutase (SOD1). To investigate the role of zebrafish ATP7A in copper homeostasis, zebrafish atp7a gene expression was reduced for analysis of downstream cellular function. The results demonstrated that zebrafish sod1 has lower expression in atp7a-knockdown fish. Similarly, zebrafish sp1, a transcriptional regulator of sod1, also shows reduced expression in atp7a-knockdown fish. The lower expression of sod1 resulting from atp7a knockdown is independent to p53 gene activation. The knockdown of atp7a and copper chelator NeoC results in hypopigmentation and notochord deformation in zebrafish. Addition of exogenous copper alleviated the impaired development. Interestingly, both sod1 and sp1 transcripts are reduced in the presence of NeoC and increased with exogenous copper, suggesting that the expression of sod1 and sp1 are directly affected by copper status. This is the first report to demonstrate a hierarchic gene expression of copper homeostatic genes between atp7a, sp1 and sod1 in zebrafish.


Effects of nicotine and tobacco-specific nitrosamines on hamster cheek pouch and gastric mucosa.
Fifty-two hamsters were divided into 6 groups and their cheek pouches treated with either 0.01% NNN, 0.01% NNN and 6% nicotine, 0.01% NNK, 0.01% NNK and 6% nicotine, 6% nicotine, or sesame oil. After painting each pouch 3 times per week for 13 months, the animals were killed and specimens from the cheek pouch and forestomach examined. Cheek pouch epithelium showed more frequent histologic changes, including hyperplasia, hyperkeratosis and, in one animal, moderate dysplasia, when nicotine was combined with NNN than after treatment with NNN (or nicotine) alone. There was a higher frequency of hyperplasia with hyperkeratosis in the forestomach when nicotine was combined with NNK than following NNK, or nicotine treatment alone, and squamous cell papillomas were evident in animals treated with both NNK and nicotine. These results suggest that in mucosal tissues nicotine may enhance the effect of weak carcinogens such as the nitrosamines.


Socio-demographic characteristics and challenges experienced by disabled patients living with HIV/AIDS in a tertiary hospital in Ibadan, Nigeria.
HIV/AIDS is highly prevalent in sub-Saharan Africa and few studies had looked at physically and mentally challenged people living with HIV/AIDS (PLHIV) in this environment. This study aimed to describe the socio-demographic characteristics and challenges faced by these patients attending University College Hospital, Ibadan. A descriptive cross-sectional study design was done. A semi-structured interviewer administered questionnaire was administered to consecutive disabled PLHIV over a period of six months. Data obtained were analyzed using descriptive and inferential statistics. A total of ninety-nine patients were seen during the study period. The mean age of these patients was 39.9 +/- 9.4 years (range 23-60 years). There were 30 (30.3%) males and 69 (69.7%) females. Twenty-four percent had no formal education while 12% had tertiary education. All respondents had suffered stigmatization/discrimination while most were poor. Eighty-seven percent had AIDS at presentation. Musculoskeletal impairments (46.5%), hearing loss (16.2%) and visual impairment (31.3%) were the commonest disability. Over twenty-three percent had prior history of road traffic accident as the cause of disability. Physically and mentally challenged people living with HIV/AIDS are poor and highly stigmatized. They require special assistance to cope and need economic empowerment to reduce their poverty level.


The Study on the Relationship between Normalized Difference Vegetation Index and Fractional Green Canopy Cover in Five Selected Crops.
Crop models are of great use and importance in modern agriculture. Most models imply spatial vegetation indices, such as NDVI, or canopy cover characteristics, such as FGCC, to provide estimation of crops conditions and forecast productivity. The purpose of the study was to (1) determine the possibility of mutual conversion between spatial NDVI and Canopeo-derived FGCC in five crops (grain corn, sunflower, tomato, millet, and winter wheat) and (2) estimate the precision of such a conversion. The data set of the study was formed by the OneSoil AI derived satellite imagery on NDVI for the studied crops in different stages of their growing season combined with Canopeo-processed photographs of vegetating crops in the field with FGCC percentage calculation. The sets of NDVI and FGCC values were paired up and then statistically processed to obtain polynomial equations of NDVI into FGCC and inverse conversion for each crop. The results of the study revealed that mutual conversion between spatial NDVI and Canopeo-derived FGCC is possible. There is a strong direct correlation (R 2 within 0.6779-0.9000 depending on the crop) between the studied indices for all crops. Close-growing crops, especially winter wheat, showed the highest correlation, while row crops and especially tomatoes had a less strong relationship between vegetation indices. The models for mutual conversion between FGCC and NDVI could be incorporated into the yield simulation models to improve the forecasting capacities.


The HNF-1alpha-SNARE connection.
Maturity-onset diabetes of the young (MODY) is a monogenic form of type 2 diabetes mellitus that is characterized by impairment of glucose-stimulated insulin secretion from pancreatic beta-cells. We previously reported that heterozygous mutations of the hepatocyte nuclear factor (HNF)-1alpha gene cause a form of MODY (MODY3). We have subsequently found that collectrin, a recently cloned kidney-specific gene of unknown function, is a novel target of HNF-1alpha in pancreatic beta-cells. In addition, we have demonstrated that collectrin forms a complex with the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex by direct interaction with snapin, a protein that is thought to be involved in neurotransmission by binding to synaptosomal-associated protein, 25 KD (SNAP25). Collectrin favours the formation of SNARE complexes and controls insulin exocytosis.


Human beta defensin-2 loaded PLGA nanoparticles impregnated in collagen-chitosan composite scaffold for the management of diabetic wounds.
Diabetic wound (DW) is the most devastating complication resulting in significant mortality and morbidity in diabetic patients. The standard treatment of DW care fails to address the prerequisites of treating DW owing to its multifactorial pathophysiology. Henceforth, developing a single treatment strategy to handle all the loopholes may effectively manage DW. The objective of the current study was to formulate Human beta defensin-2 (HBD-2) loaded Poly (lactic-co-glycolic acid) (PLGA) nanoparticle impregnated in collagen/chitosan (COL-CS) composite scaffolds for the accelerated healing of DW. Upon investigation, the developed biodegradable crosslinked scaffold possesses low matrix degradation, optimum porosity, and sustained drug release than the non-crosslinked scaffold. In vitro studies revealed that the HBD-2 COL-CS scaffold was biocompatible and accelerated cell migration and angiogenesis. The HBD-2 COL-CS scaffold showed significant antimicrobial activity in S. aureus, E. coli, and P. aeruginosa. The in vivo studies revealed that the HBD-2 COL-CS treated group accelerated healing compared to those in COL-CS and control groups. The ELISA results indicated a significant decrease in MMP-9, TNF-α, MPO, NAG, and NO with an increase in IL-10 in HBD-2 COL-CS treated group. The accelerated healing in HBD-2 COL-CS treated group might be due to the synergistic effects of PLGA (collagen synthesis and deposition and positive angiogenic effect), HBD-2 (anti-inflammatory, antibacterial, positive angiogenic effect, cell proliferation, and migration), COL (established wound healer and stabilizer) and CS (antibacterial, controlled drug release).


Medial branch blocks versus pericapsular blocks in selecting patients for percutaneous cryodenervation of lumbar facet joints.
At many institutions, it is not practically feasible to perform a series of controlled or placebo-controlled medial branch blocks on several facet joints in order to select patients for facet joint rhizotomy. As for uncontrolled blocks, there is no proof that medial branch blocks are superior to other types of blocks. This study was performed to compare medial branch blocks to simple pericapsular blocks for the selection of patients for lumbar facet joint cryodenervation. Patient selection was based on history, imaging, and physical examination. Diagnostic blocks were either medial branch blocks or pericapsular blocks. Percutaneous medial branch cryodenervation was performed by use of a Lloyd Neurostat 2000. Outcome parameters were low back pain (visual analog scale [VAS]), limitation of activity (Macnab), and overall satisfaction. A total of 26 patients were recruited, 13 for each group. Follow-up was 6 months. Patients who had been selected by medial branch blocks had better pain relief than did patients who had been diagnosed by use of pericapsular blocks. At 6 weeks and at 3 months after treatment, these results reached statistical significance (VAS 2.2 v 4.2, P < .05). Our results suggest that uncontrolled medial branch blocks are superior to pericapsular blocks in selecting patients for facet joint cryodenervation, but both blocks work. If serial controlled blocks cannot be used, lumbar facet joint pain remains a diagnostic dilemma.


Posterior Humeral Avulsion of the Glenohumeral Ligament and Associated Injuries: Assessment Using Magnetic Resonance Imaging.
Lesions associated with posterior humeral avulsion of the glenohumeral ligament (HAGL) can lead to persistent symptoms related to posterior shoulder instability and can be commonly missed or delayed in diagnosis. To identify and characterize the MRI findings in patients with a posterior HAGL lesion. Case series; Level of evidence, 4. This retrospective case series included 27 patients (28 shoulders) identified by search through the senior authors' databases, with cross-reference to their institutional radiologic communication system for MRI review. Baseline patient demographic data were collected, including age and sex. All posterior HAGL lesions were identified on MRI and characterized as partial, complete, or floating lesions. All acute glenohumeral pathologic changes concurrent with the posterior HAGL were documented. Chondrolabral retroversion of the injured shoulder was measured on axial MRI. The average age of the identified cohort was 33.6 years (range, 15-81 years), and 23 patients were male (86%). Posterior HAGL injuries were found to be complete tears (71%), partial tears (25%), and floating lesions (4%); concomitant bony HAGL avulsion was found in 7% of injuries. Additional traumatic glenohumeral disorders occurred in 93% of cases. The most common concurrent injuries were reverse Hill-Sachs lesions (36%), anterior Bankart lesions (29%), and posterosuperior rotator cuff tears (25%). Notably, concomitant anterior labral or capsular injury was found in 50% of patients, signifying bidirectional disruption of the capsule. In addition, increased chondrolabral version was found in this cohort (10.2° ± 3.7° retroversion). This study depicts the high association of combined injury with posterior HAGL lesions and increased chondrolabral retroversion. Findings on MRI related to a posterior HAGL injury could potentially be masked by additional injury and may occur with mechanisms that also lead to anterior glenohumeral disorders.


The accuracy of general practitioner workforce projections.
Health workforce projections are important instruments to prevent imbalances in the health workforce. For both the tenability and further development of these projections, it is important to evaluate the accuracy of workforce projections. In The Netherlands, health workforce projections have been done since 2000 to support health workforce planning. What is the accuracy of the techniques of these Dutch general practitioner workforce projections? We backtested the workforce projection model by comparing the ex-post projected number of general practitioners with the observed number of general practitioners between 1998 and 2011. Averages of historical data were used for all elements except for inflow in training. As the required training inflow is the key result of the workforce planning model, and has actually determined past adjustments of training inflow, the accuracy of the model was backtested using the observed training inflow and not an average of historical data to avoid the interference of past policy decisions. The accuracy of projections with different lengths of projection horizon and base period (on which the projections are based) was tested. The workforce projection model underestimated the number of active Dutch general practitioners in most years. The mean absolute percentage errors range from 1.9% to 14.9%, with the projections being more accurate in more recent years. Furthermore, projections with a shorter projection horizon have a higher accuracy than those with a longer horizon. Unexpectedly, projections with a shorter base period have a higher accuracy than those with a longer base period. According to the results of the present study, forecasting the size of the future workforce did not become more difficult between 1998 and 2011, as we originally expected. Furthermore, the projections with a short projection horizon and a short base period are more accurate than projections with a longer projection horizon and base period. We can carefully conclude that health workforce projections can be made with data based on relatively short base periods, although detailed data are still required to monitor and evaluate the health workforce.


[An almost fatal airway obstruction caused by a secretion plug in an endotracheal tube].
Airway obstruction caused by a secretion plug in an endotracheal tube or a tracheostomy cannula can be a serious complication to mechanical ventilation. This case describes an event caused by a mucus plug localized to the distal part of a tracheostomy tube in a tetraplegic patient. The plug functioned as a check valve, allowing air to pass in - but not out - of the patient. A suction catheter could be passed through the airway without any resistance, and thus the airway was believed to be free. The event, which had an almost fatal outcome due to hyperinflation of the lungs, was relieved by replacement of the tracheostomy cannula.


CYP2D6 genetic polymorphisms in Southern Mexican Mayan Lacandones and Mestizos from Chiapas.
In previous CYP2D6 genotyping studies in Mexican-Amerindians a very low frequency of poor metabolizers (PMs) has been reported. Moreover, ultrarapid metabolizers (UMs) status has only been analyzed in some groups from Northern Mexico. In the present study we evaluated the hypothesis of low frequency of PMs in Mexican-Amerindians in Southern Mexican populations from Chiapas (Lacandones [ML] vs Mestizos [MM]). The frequency of UMs is also reported. CYP2D6 alleles *2, *3, *4, *5, *6, *10, *17, *35 and *41 and copy number variations were analyzed in 154 ML and 100 MM healthy volunteers. The PM frequency was 0% in MLs and 1% in MMs, and for UMs was 2.6% in MLs and 3% in MMs. The present data support previous findings reporting a very low frequency of CYP2D6 PMs in Mexican-Amerindians. Furthermore, the predicted UM phenotype in both MMs and MLs was lower than those reported for most Mexican populations.


α2δ expression sets presynaptic calcium channel abundance and release probability.
Synaptic neurotransmitter release is driven by Ca(2+) influx through active zone voltage-gated calcium channels (VGCCs). Control of active zone VGCC abundance and function remains poorly understood. Here we show that a trafficking step probably sets synaptic VGCC levels in rats, because overexpression of the pore-forming α1(A) VGCC subunit fails to change synaptic VGCC abundance or function. α2δs are a family of glycosylphosphatidylinositol (GPI)-anchored VGCC-associated subunits that, in addition to being the target of the potent neuropathic analgesics gabapentin and pregabalin (α2δ-1 and α2δ-2), were also identified in a forward genetic screen for pain genes (α2δ-3). We show that these proteins confer powerful modulation of presynaptic function through two distinct molecular mechanisms. First, α2δ subunits set synaptic VGCC abundance, as predicted from their chaperone-like function when expressed in non-neuronal cells. Second, α2δs configure synaptic VGCCs to drive exocytosis through an extracellular metal ion-dependent adhesion site (MIDAS), a conserved set of amino acids within the predicted von Willebrand A domain of α2δ. Expression of α2δ with an intact MIDAS motif leads to an 80% increase in release probability, while simultaneously protecting exocytosis from blockade by an intracellular Ca(2+) chelator. α2δs harbouring MIDAS site mutations still drive synaptic accumulation of VGCCs; however, they no longer change release probability or sensitivity to intracellular Ca(2+) chelators. Our data reveal dual functionality of these clinically important VGCC subunits, allowing synapses to make more efficient use of Ca(2+) entry to drive neurotransmitter release.


Microscopy and Image Analysis.
This unit provides an overview of light microscopy, including objectives, light sources, filters, film, and color photography for fluorescence microscopy and fluorescence in situ hybridization (FISH). We believe there are excellent opportunities for cytogeneticists, pathologists, and other biomedical readers, to take advantage of specimen optical clearing techniques and expansion microscopy-we briefly point to these new opportunities. © 2017 by John Wiley & Sons, Inc.


Influence of gender and socioeconomic background on the decision to perform gallstone surgery: a population-based register study.
In the absence of unequivocal standardized indications for surgery, socioeconomic background and gender may have a major impact on the decision to perform surgery for cholecystolithiasis. The purpose was to assess how decisions to perform surgery in Sweden are influenced by patient-related factors and how this affects the epidemiology of gallstone disease. This study is based on the Swedish Register for Gallstone Surgery and Endoscopic Retrograde Cholangiopancreatography (GallRiks), which covers >90% of surgical units, including 98% of all procedures performed. All procedures performed during 2005-2009 were included. Data on socioeconomic background were obtained from Statistics Sweden. The influence of gender and age on decision to perform surgery was tested in multivariate linear regression analysis. Up to the age of 40 years, women were 6 times more likely than men to undergo surgery for biliary colic. On the other hand, there was a relative preponderance of men undergoing cholecystectomy for jaundice, cholecystitis, bile duct stone or pancreatitis in the elderly population (p < 0.001). Socioeconomic background did not have any significant impact on the decision to operate. Presentations of gallstone disease differ between men and women, as does the decision to perform surgery. The higher incidence of surgery for secondary complications in older men could be explained by a higher prevalence of gallstones resulting from a lower incidence of surgery at a younger age. Whether or not wider indications for surgery in young patients reduce the risk for gallstone complications requiring surgery should be explored in future studies.


Unprecedented spin localisation in a metal-metal bonded dirhenium complex.
The molecular and electronic structure of edge-sharing bioctahedral [N(n-Bu)4]3[Re2(mnt)5] is reported here. Despite the short intermetal bond length of 2.6654(2) Å with computed bond order of 1.2, the unpaired electron is localised by the asymmetric ligand distribution, as demonstrated by its remarkable EPR spectrum.


The road to the winner's circle begins at the ASHRM Annual Conference!
The saying "Time flies when you're having fun" is a time-honored expression that is part of our vernacular. Sometimes we say it in jest, tongue in cheek, when it seems a long day will never end! However, when we celebrate professional or personal anniversaries, when we look at our growing kids, or especially, when we look in the mirror, we marvel at how fast the years have flown by!


Polysulfone corneal lenses.
Polysulfone is a thermoplastic compound first synthesized in 1965. A unique characteristic of polysulfone is its high refractive index (1.633), which allows very thin optical lenses to be manufactured. Over the last five years, D. Peter Choyce has surgically implanted over 40 polysulfone lenses into eyes of his patients. Analysis of his data indicates that polysulfone intraocular lenses are capable of correcting large refractive errors. Based on his work, a multicenter study was undertaken to evaluate the safety and efficacy of polysulfone as an intracorneal lens material in laboratory models. Four monkeys, eight baboons, and 24 cats were used as laboratory models; 5.0-mm to 6.0-mm diameter hyperopic (+28.5 diopter) and myopic (-17.0 and -25.5 diopter) lenses were surgically implanted within the corneal stroma in one eye of each of the laboratory models while a sham lamellar dissection was performed in the other eye. One hundred percent (4/4) of monkey eyes, 12.5% (1/8) of baboon eyes, and 70.0% (18/24) of cat eyes maintained clear media by ophthalmoscopic examination at follow-ups ranging from three to six months. Complications included both visually and nonvisually significant interface opacities, lens extrusion, anterior corneal necrosis, refractile particles, and epithelial thinning.


Next generation RNA-sequencing in prognostic subsets of chronic lymphocytic leukemia.
Advances in next-generation RNA-sequencing have revealed the complexity of transcriptomes by allowing both coding and noncoding(nc)RNAs to be analyzed. However, limited data exist regarding the whole transcriptional landscape of chronic lymphocytic leukemia(CLL). In this pilot-study, we evaluated RNA-sequencing in CLL by comparing two subsets which carry almost identical or "stereotyped" B-cell receptors with distinct clinical outcome, that is the poor-prognostic subset #1 (n 5 4) and the more favorable-prognostic subset #4(n 5 4). Our analysis revealed that 156 genes (e.g. LPL, WNT9A) and 76 ncRNAs, (e.g. SNORD48, SNORD115) were differentially expressed between the subsets. This technology also enabled us to identify numerous subset-specific splice variants (n 5 406), which were predominantly expressed in subset #1, including a splice-isoform of MSI2 with a novel start exon. A further important application of RNA-sequencing was for mutation detection and revealed 16–30 missense mutations per sample; notably many of these changes were found in genes with a strong potential for involvement in CLL pathogenesis, e.g., ATM and NOTCH2.This study not only demonstrates the effectiveness of RNA-sequencing for identifying mutations, quantifying gene expression and detecting splicing events, but also highlights the potential such global approaches have to significantly advance our understanding of the molecular mechanisms behind CLL development.


Alterations in fetal kidney development and elevations in arterial blood pressure in young adult sheep after clinical doses of antenatal glucocorticoids.
Epidemiologic studies have yielded controversial information regarding an association between antenatal steroid administration and elevations in arterial blood pressure (BP). The aim of the study was to determine whether antenatal administration of a clinically relevant dose of steroids at a time when fetal nephrogenesis is at its highest results in abnormal kidney development and adult hypertension. Pregnant sheep were treated with either vehicle or betamethasone. Maternal injections were given 24 h apart at 80 d of gestational age (dGA; 0.55 of gestation). Animals were studied either as fetuses or as immature adults. Fetuses were delivered by cesarean section at 135 dGA. Adults were studied at 6 mo of age. Betamethasone administration did not induce premature labor or intrauterine growth restriction. In the betamethasone-exposed group, we found at 135 dGA a 25.5% decrease in the number of glomeruli with no differences in fetal kidney weight. In adults, mean, systolic, and diastolic arterial BPs were significantly higher, whereas there were no significant differences in heart rate over the same study period. The major finding of this study is that a single course of antenatal steroids alters renal development and is associated with elevations in arterial BP in lambs at 6 mo of age. We conclude that antenatal glucocorticoid administration under the National Institutes of Health consensus guidelines may alter human fetal renal development.


High prevalence of West Nile virus: a continuing risk in acquiring infection from a mosquito bite.
The prevalence of West Nile Virus (WNV) was evaluated by diplex real-time RT-PCR assay for the years 2001-2005 in Culex species of mosquitoes, several species of dead birds, and clinically suspected mammals collected in Kansas. The analysis was performed using a TaqMan-based diplex real-time RT-PCR assay targeted against two regions of the WNV genome, envelope glycoprotein gene and 3' untranslated region. The assay aided in the accurate detection of WNV in mosquitoes at high prevalence for the years 2002-2005. Similarly, high incidence of birds that tested positive for WNV was detected in 2002-2004. WNV positives in mammals by the diplex real time RT-PCR assay included horses, squirrels, mules, sheep and a mountain goat. Majority of the equine WNV positives were detected only in the year 2002. Sequence analysis of a segment of the envelope glycoprotein gene from 31 randomly selected WNV positive samples revealed variations in six samples at one or two nucleotide positions. The identity of high levels of WNV positives in Kansas parallels the recent reports on the widespread distribution of the virus in the United States. The continued detection of WNV in the mosquitoes is of significant public health concern and calls for continued surveillance and public health activities.


Antidepressant effects of ERbeta-selective estrogen receptor modulators in the forced swim test.
Estradiol (E2) may influence depressive symptomology of women and decrease depressive behavior among rodents. The mechanism(s) for E2's antidepressant effects are not well understood. To determine whether antidepressant effects of E2 may involve actions at intracellular estrogen receptor (ER) alpha or beta isoforms, selective ER modulators (SERMs) were administered (10 microg sc) to ovariectomized rats 48 h before testing in the forced swim test, an animal model of depression, and the horizontal crossing task. Rats received sesame oil vehicle, 17beta-E2, which has a high affinity for ERalpha and ERbeta, SERMs that vary in their activity at ERalpha and beta, or a tricyclic antidepressant (desipramine; 30 mg/kg ip), as a positive control. ERalpha-selective SERMs were propyl pyrazole triol (PPT) and 17alpha-E2. PPT has more selective effects at ERalpha than does 17alpha-E2, which also binds ERbeta. ERbeta-selective SERMs were diarylpropionitrile (DPN) and 7,12-dihydrocoumestan (coumestrol). DPN is more selective at ERbeta than coumestrol, which also binds ERalpha. 17beta-E2, ERbeta-selective SERMs (DPN, coumestrol), and desipramine administration produced antidepressive behavior (decreased immobility, increased struggling and swimming). ERalpha-selective SERMs (PPT, 17alpha-E2) were not different from vehicle. There were no differences among groups in the number of beam breaks made in the horizontal crossing task. These data suggest that E2's antidepressive effects may involve actions at ERbeta.


In vitro inhibition of the infectivity and replication of human immunodeficiency virus type 1 by combination of antiretroviral 2',3'-dideoxynucleosides and virus-binding inhibitors.
We tested the in vitro inhibitory activities of three 2',3'-dideoxynucleosides and two inhibitors of viral binding in combinations against the infectivity and cytopathic effect of human immunodeficiency virus type 1. 3'-Azido-2',3'-dideoxythymidine, 2',3'-dideoxyinosine, or 2',3'-dideoxycytidine, combined with recombinant soluble CD4 (sCD4), brought about synergistic antiretroviral activity without toxicity at clinically achievable concentrations. Combinations of 2',3'-dideoxynucleosides plus dextran sulfate exerted similar synergistic antiviral effects without concomitant increases in toxicities. When sCD4 and dextran sulfate were combined, apparent antagonism was observed. We confirmed that no combination of sCD4 plus 3'-azido-2',3'-dideoxythymidine, 2',3'-dideoxyinosine, or 2',3'-dideoxycytidine significantly increased the inhibitory effect on colony formation of human myeloid-monocytic bone marrow cells in vitro at the concentrations used in this study. These data might have clinical relevance for the treatment of patients infected with human immunodeficiency virus.


Growth and nitrogen retention of children consuming all of the day's protein intake in one meal.
To ascertain that linear growth and weight gain continue while consuming a diet in which all the day's protein is given in one meal, six children (ages: 8.5 to 25 months, weight: 6 to 8 kg) who were convalescing from malnutrition were placed on such a diet for 60 to 67 days. All diets provided 125 to 150 kcal and 2.0 to 2.5 g protein/kg/day, recalculated weekly. Energy was evenly divided among five feedings but the entire day's protein was given at the third meal. Increases in height age relative to increases in chronological age were appropriate for the stage of recovery. Height quotient (height age/chronological age times 100) increased during the study in four children and remained constant in two. Increases in weight age were greater than corresponding increases in chronological age. Nine-day control periods prior to and after the growth study allowed comparative N balances, which confirmed previous findings that apparent N retention from the unevenly distributed protein (32 +/- 10% of intake) was less, although not significantly so, than that from the control diet (34 +/- 15% of intake). Fecal facts were elevated during the study period (18 +/- 7.5% of intake) relative to the control period values (9 +/- 3.9% of intake) in all but one child.


Contralateral facial nerve palsy following mandibular second molar removal: is there co-relation or just coincidence?
Peripheral facial nerve palsy (FNP) is the most common cranial nerves neuropathy. It is very rare during dental treatment. Classically, it begins immediately after the injection of local anaesthetic into the region of inferior dental foramen and it's homolateral to the injection. Recovery takes a few hours, normally as long the anaesthetic lasts. The authors present a 44-year-old patient who presented a contralateral delayed-onset facial paralysis arising from dental procedure and discuss the plausible pathogenesis mechanism of happen and a possible relationship between dental procedure and contralateral FNP.


Age-related Macular Degeneration: Current Knowledge of Zinc Metalloproteinases Involvement.
Advanced age-related macular degeneration (AMD) is the leading cause of blindness in the elderly with limited therapeutic options. The disease is characterized by photoreceptor loss in the macula and reduced Retinal Pigment Epithelium (RPE) function, associated with matrix degradation, cell proliferation, neovascularization and inflammation. Matrix metalloproteinases (MMPs), a disintegrin and metalloproteinases (ADAMs) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) play a critical role in the physiology of extracellular matrix (ECM) turnover and, in turn, in ECM pathologies, such as AMD. A balance between the activities of MMPs and Tissue Inhibitors of Metalloproteinase (TIMPs) is crucial for the integrity of the ECM components; indeed, a dysregulation in the ratio of these factors produces profound changes in the ECM, including thickening and deposit formation, which eventually might lead to AMD development. This article reviews the relevance and impact of zinc metalloproteinases on the development of AMD and their roles as biomarkers and/or therapeutic targets. We illustrate some studies on several inhibitors of MMPs currently used to dissect physiological properties of MMPs. Moreover, all molecules or technologies used to control MMP and ADAM activity in AMD are analyzed. This study underlines the changes in the activity of MMPs expressed by RPE cells, highlights the functions of already used MMP inhibitors and consequently suggests their application as therapeutic agents for the treatment of AMD.


Pulse methylprednisolone therapy for the treatment of extensive alopecia areata.
Since the mainstay of pathogenesis depends on autoimmune process, systemic steroids are widely used in the treatment of alopecia with various side effects. To avoid side effects of long-term steroid treatment, pulse methylprednisolone therapy appears to be a safe treatment option. The aim was to determine the effect of pulse methylprednisolone therapy for the treatment of adult alopecia areata. Demographical features of all patients were recorded before the treatment. Patients received methylprednisolone 500 mg intravenously for 3 consecutive days every month for 3 months. Patients were followed up for 3 months. Treatment responses were defined by complete regrowth (100%), significant regrowth (>50%) and minimal regrowth (<50%). Totally 15 patients were enrolled in this study. At the end of the study, two patients had significant regrowth and one patient had minimal regrowth in multifocal alopecia areata (n = 4); one patient had significant regrowth and one patient had minimal regrowth in alopecia universalis (n = 8); three patients had no regrowth in alopecia totalis (n = 3). The study suggests that pulse methylprednisolone therapy might be a therapeutic option for severe multifocal alopecia areata. However, in alopecia totalis or universalis, treatment results are unsatisfactory.


Gender, Competitiveness, and Intentions to Pursue STEM fields.
Building on the insights of gender theorists as well as a small body of emerging quantitative research, the authors examine whether and how students' self-perceptions of competitiveness are related to gendered patterns of future STEM expectations among a sample of U.S. high school students. Results of regression analyses reveal that female students' relatively lower self-perceptions of competitiveness (compared to male students) significantly contribute to their lower expectation of majoring in two historically male-dominated fields, physical science and engineering. Additional results revealed an interaction between gender and competitiveness for expectations to major in computer science, such that while girls' expectations significantly increase with their perceptions of competitiveness, boys' decisions to pursue computer science are unrelated to such perceptions.


Electrochemical synthesis and structural characterization of a novel mixed-valence copper(I)-copper(II) complex: {[bis(ethylenediamine)copper(II)] bis[diiodocuprate(I)]}.
A novel, mixed-valent copper(I)-copper(II) complex, {[bis(ethylene-diamine)copper(II)] bis[diiodocuprate(I)]} (1), has been prepared by electrochemical dissolution of a sacrificial copper anode in a solution of ethylenediamine (en), I2 and tetraethylammoniumperchlorate (TEAP) as supporting electrolyte in acetonitrile (AcN)and characterized by single-crystal X-ray structure determination. The crystal structure of the complex 1 shows that it consists of a CuI2 polymer formed from I- ligands bridging Cu(I) ions, with a nearly square planar geometry of bivalent Cu(II) atoms chelated by two ethylenediamine ligands. The results also show that direct electrosynthesis of the complex had high current efficiency, purity and electrolysis yield.


Comparison of the effects between animal-derived trypsin and recombinant trypsin on human skin cells proliferation, gene and protein expression.
Animal-derivative free reagents are preferred in skin cell culture for clinical applications. The aim of this study was to compare the performance and effects between animal-derived trypsin and recombinant trypsin for skin cells culture and expansion. Full thickness human skin was digested in 0.6 % collagenase for 6 h to liberate the fibroblasts, followed by treatment with either animal-derived trypsin; Trypsin EDTA (TE) or recombinant trypsin; TrypLE Select (TS) to liberate the keratinocytes. Both keratinocytes and fibroblasts were then culture-expanded until passage 2. Trypsinization for both cell types during culture-expansion was performed using either TE or TS. Total cells yield was determined using a haemocytometer. Expression of collagen type I, collagen type III (Col-III), cytokeratin 10, and cytokeratin 14 genes were quantified via RT-PCR and further confirmed with immunocytochemical staining. The results of our study showed that the total cell yield for both keratinocytes and fibroblasts treated with TE or TS were comparable. RT-PCR showed that expression of skin-specific genes except Col-III was higher in the TS treated group compared to that in the TE group. Expression of proteins specific to the two cell types were confirmed by immunocytochemical staining in both TE and TS groups. In conclusion, the performance of the recombinant trypsin is comparable with the well-established animal-derived trypsin for human skin cell culture expansion in terms of cell yield and expression of specific cellular markers.


Digital image processing of crystalline specimens examined by electron microscopy.
Crystalline specimens imaged in the electron microscope are analysed using digital processing. Some principles of structural analysis using the method of Fourier decomposition are discussed. Complementary techniques, such as enhancement by gradient and Laplacian operators, have been found useful in analysing electron micrographs. The application of these techniques to some problems in Materials Science and Biology are reviewed. By selecting and phase-correcting spots in the computed diffraction pattern, it was possible to localize atoms, molecules, and their defective arrangement in evaporated gold, sputter-deposited tungsten films, and single crystals of cadmium selenide. Digital processing based on the theory of helical diffraction was used to explore the three-dimensional arrangement of molecules in cellular components of alveolar soft part sarcoma, Hirano bodies, and neurofibrillar tangles in the human brain.


Microstructure and Mechanical Properties of 34CrNiMo6 Steel Repaired by Friction Stir Processing.
Repairing damaged parts using proper repairing methods has become an important means to reduce manufacturing and operational costs and prolong the service life of 34CrNiMo6 steel structures. In the conventional fusion repairing method, welding wire and powder are often used as filling materials. Filling materials are often expensive or difficult to find. Some metallurgical issues (such as solidification crack, higher distortion) were also found with these methods. At the same time, most of the equipment that requires welding wire and powder is expensive. In this study, a new method based on friction stir processing (FSP) was successfully employed to repair 34CrNiMo6 steel, using a block as filling material. Filling blocks are much cheaper than conventional fusion repair consumables. As a result of solid-state repair, this method can also avoid the metallurgical issues of fusion repair. The microstructure and mechanical properties of the repaired samples were investigated using OM (Optical Microscope), SEM, EDS (Energy Dispersive Spectroscopy), XRD, and a Vickers hardness electronic universal tensile tester. The results showed that 34CrNiMo6 steel was successfully repaired by this method, with no defect. Tensile tests showed that the maximum ultimate strength (UTS) was 900 MPa and could reach 91.8% of that of the substrate. The fracture mode of the tensile samples was ductile/brittle mixed fracture. Hence, the repairing method based on FSP appears to be a promising method for repairing castings.


Engineered cationic antimicrobial peptides to overcome multidrug resistance by ESKAPE pathogens.
Multidrug resistance constitutes a threat to the medical achievements of the last 50 years. In this study, we demonstrated the abilities of two de novo engineered cationic antibiotic peptides (eCAPs), WLBU2 and WR12, to overcome resistance from 142 clinical isolates representing the most common multidrug-resistant (MDR) pathogens and to display a lower propensity to select for resistant bacteria in vitro compared to that with colistin and LL37. The results warrant an exploration of eCAPs for use in clinical settings.


Increasing arsenic accumulation as an implication of climate change: a case study using red algae.
Climate change due to an increasing concentration of carbon dioxide in the atmosphere is a global issue. It can impact aquatic environments by affecting water flow, pollutant transformation and migration, and other toxicant-related effects. We assessed the interactive effects of temperature warming and pH changes on variations in accumulation of total arsenic (AsT) in the red alga Sarcodia suae at different levels of arsenite (AsIII). Result showed that AsT variations in the alga were moderated by significant joint effects of warming temperature and/or increasing pH levels and their interactions with increasing AsIII concentrations. Our study suggests possible deleterious impacts on macroalgal populations due to toxicological effects associated with prevailing environmental conditions. Therefore, improved pollution management, climate change adaptation, and mitigation strategies are needed to deal with current environmental issues and As aggravation.


Correction: Anti-osteosarcoma effect of hydroxyapatite nanoparticles both in vitro and in vivo by downregulating the FAK/PI3K/Akt signaling pathway.
Correction for 'Anti-osteosarcoma effect of hydroxyapatite nanoparticles both in vitro and in vivo by downregulating the FAK/PI3K/Akt signaling pathway' by Renxian Wang et al., Biomater. Sci., 2020, 8, 4426-4437, DOI: 10.1039/D0BM00898B.