Experience-dependent neural substrates involved in vocal pitch regulation during singing.
Proper singing requires the integration of auditory feedback mechanisms with the vocal motor system, such that vocal pitch can be precisely controlled. To determine the neural substrates involved in audio-vocal integration, non-musicians and experienced singers underwent fMRI scanning while they sang a single tone with either unaltered ("simple") or pitch-shifted auditory feedback; in pitch-shifted trials, subjects were instructed either to ignore or compensate for the shifted feedback. We hypothesized that the anterior cingulate cortex (ACC), superior temporal gyrus (STG), and anterior insula may be involved in audio-vocal integration due to their functional roles during singing and their anatomical connectivity. Although singers were more accurate than non-musicians in simple singing, both groups recruited similar functional networks. Singers ignored the shifted feedback better than non-musicians, and both groups also displayed different patterns of neural activity for this task: singers recruited bilateral auditory areas and left putamen, while non-musicians recruited the left supramarginal gyrus and primary motor cortex. While there were no significant group differences in performing the compensate task, singers displayed enhanced activity in the ACC, superior temporal sulcus, and putamen, whereas non-musicians exhibited increased activity in the dorsal premotor cortex, a region involved with sensorimotor interactions. We propose two neural substrates for audio-vocal integration: the dorsal premotor cortex may act as a basic interface, but with vocal training and practice, the ACC, auditory cortices, and putamen may be increasingly recruited as people learn to monitor their auditory feedback and adjust their vocal output accordingly.


Risk factors for unfavorable outcome of pulmonary tuberculosis in adults in Taipei, Taiwan.
This study was undertaken to identify factors associated with unfavorable outcomes in patients with pulmonary tuberculosis (PTB) in Taipei, Taiwan in 2007-2008. Taiwanese adults with culture-positive PTB diagnosed in Taipei during the study period were included in this retrospective cohort study. Unfavorable outcomes were classified as treatment default, death, treatment failure, or transfer. Of 1616 eligible patients, 22.6% (365) had unfavorable outcomes, mainly death. After controlling for patient sociodemographic factors, clinical findings, and underlying disease, independent risk factors for unfavorable outcomes included advanced age, unemployment, end-stage renal disease requiring dialysis, malignancy, acid-fast bacilius smear-positivity, multidrug-resistant TB, and notification from ordinary ward or intensive care unit. In contrast, patients receiving directly observed treatment, and with a high school or higher education were significantly less likely to have unfavorable outcomes. This study advanced our understanding by revealing that a high school or higher education might lower the risk of an unfavorable outcome. Our results also confirmed the risk factors for unfavorable outcomes shown in previous research. Future TB control programmes in Taiwan should target particularly high-risk patients including those who had lower educational levels.


In Situ Detection of Water Leakage for Textile-Reinforced Composites.
By incorporating electrically conductive yarns into a waterproof membrane, one can detect epoxy resin cracking or liquid leakage. Therefore, this study examined the electrical conductivity variations of several yarns (metallic or carbon-based) for cracking and water detection. The first observations concerned the detectors' feasibility by investigating their conductivity variations during both their resin implementation processes and their resin cracking. Throughout this experiment, two phenomena were detected: the compression and the separation of the fibres by the resin. In addition, the resin cracking had an important role in decreasing the yarns' conductivity. The second part of this study concerned water detection. Two principles were established and implemented, first with yarns and then with yarns incorporated into the resin. First, the principle of absorption was based on the conductivity variation with the yarns' swelling after contact with water. A short circuit was established by the creation of a conductive path when a drop of water was deposited between two conductive, parallel yarns. Through the influence of the yarns' properties, this study explored the metallic yarns' capacity to better detect water with a short circuit and the ability of the carbon-based yarns to detect water by the principle of absorption.


The chemical inducer, BTH (benzothiadiazole) and root colonization by mycorrhizal fungi (Glomus spp.) trigger resistance against white rot (Sclerotinia sclerotiorum) in sunflower.
White rot caused by Sclerotinia sclerotiorum (SS) is one of the most devastating plant diseases of sunflower. Controlling this pathogen by available tools hardly result in acceptable control. The aim of this study was to elucidate the effects of plant resistance inducers, BTH (benzothiadiazole in Bion 50 WG) and arbuscular mycorrhizal fungi (AMF) on disease development of white rot in three sunflower genotypes. Defence responses were characterized by measuring the disease severity and identifying cellular/histological reactions (e.g. autofluorescence) of host plants upon infection. Depending on the host genotype, a single application of inducers reduced disease symptoms. Histological examination of host responses revealed that BTH and/or AMF pre-treatments significantly impeded the development of pathogenic hyphae in Iregi szürke csíkos and P63LE13 sunflower plants and it was associated with intensive autofluorescence of cells. Both localized and systemic induction of resistance was observed. Importantly, the frequency of mycorrhization of hybrid P63LE13 and PR64H41 was significantly increased upon BTH treatment, so it had a positive effect on the formation of plant-mycorrhiza interactions in sunflower. To our knowledge, this is the first report on the additive effect of BTH on mycorrhization and the positive effect of these inducers against SS in sunflower.


Emergency presentation of cystic lymphangioma of the colon: A case report and literature review.
Colonic lymphangioma is an unusual benign malformation. The clinical presentation of lymphangiomas vary from incidental discovery on imaging to presenting with acute abdomen. We present the case of a 73-year-old male, undergoing surgery due to acute abdomen associated with severe anemia, in whose case a lymphangioma of the cecum was recognised only in the postoperative histopathological examination. The management of colonic lymphangioma depends on the individual situation; close surveillance or endoscopic therapy may be appropriate for asymptomatic lesions smaller than 2.5cm in diameter. Surgical intervention can be considered for larger lesions or in patients who develop complications. The interest in our case lies in the relatively rare diagnosis of colon lymphangioma and how the cardinal sign was anemia, which may be due to the serosanguineous cystic contents of lymphangiomatosis from the internal bleeding and can cause fatal complications that require emergency surgery.


Clinical significance of novel biomarkers to predict the natural course of hepatitis B infection.
Chronic hepatitis B (CHB) can be divided into immune tolerance (IT), immune clearance (IC), hepatitis B e antigen (HBeAg)-negative inactive/quiescent carrier (ENQ), and HBeAg-negative hepatitis (ENH) phases. The conventional biomarkers used to distinguish these phases have limitations. We examined the clinical significance of hepatitis B virus (HBV) RNA and hepatitis B core-related antigen (HBcrAg) as novel biomarkers. One hundred eighty-nine patients without treatment currently were categorized by CHB phase (IT = 46, IC = 45, ENQ = 49, ENH = 49). The associations of HBV RNA and HBcrAg with HBV DNA and alanine transaminase (ALT) were analyzed. The decision tree model was used to distinguish the four phases in the natural course of CHB. The concentrations of HBV RNA and HBcrAg were highest in the IT and IC phases (P < 0.01). Serum HBV RNA was similar to HBcrAg in treatment-naïve patients. HBV RNA and HBcrAg correlated with HBV DNA in the HBeAg+ and HBeAg- status (HBV RNA: e+ r = 0.51, e- r = 0.62; HBcrAg: e+ r = 0.51, e- r = 0.71), but their association with HBV DNA differed among phases. The accuracy, sensitivity, and specificity of HBcrAg with ALT in distinguishing the CHB phases were 95.65%, 95.83%, and 95.55%, respectively. Serum HBV RNA and HBcrAg may be useful to monitor CHB progression.


Persistent transgene expression following intravenous administration of a liposomal complex: role of interleukin-10-mediated immune suppression.
Studies conducted in non-tumor-bearing, immunocompetent mice have shown that intravenous administration of liposome-DNA complex elicits an inflammatory response that results in a failure to sustain adequate transgene expression. In the present study, however, we investigated the effects of a cationic liposomal DOTAP:cholesterol (DOTAP:Chol)-DNA complex on cytokine production and transgene expression in both experimental lung tumor-bearing (TB) mice and non-tumor-bearing (NTB) syngeneic mice and nude mice. Intravenous injection of DOTAP:Chol-luciferase (luc) DNA complex resulted in tumor necrosis factor-alpha levels that were 50% lower and interleukin-10 levels that were 50-60% higher in TB mice than in NTB mice. Furthermore, a significant increase in luc expression (P = 0.001) that persisted for 7 days was observed in TB mice. In contrast, luc expression decreased significantly from day 1 to day 2 in NTB mice. Also, luc expression was two- to threefold higher in TB mice that were given multiple injections of DOTAP:Chol-luc complex than in mice who received a single injection. In contrast, luc expression was significantly suppressed following multiple injections in NTB mice (P = 0.01). Further analysis revealed IL-10 protein expression by the tumor cells in TB mice. Injection of anti-IL-10 antibody in TB mice resulted in a significant decrease in luc expression (P = 0.01) compared with that in mice injected with a control antibody. Based on these findings, we conclude that transgene expression persists in TB mice and is partly mediated by IL-10. Additionally, multiple injections of liposome-DNA complex can increase transgene expression in TB mice. These findings have clinical applications in the treatment of cancer.


Isotherm and kinetic investigations of sawdust-based biochar modified by ammonia to remove methylene blue from water.
Chemical industry effluent may pose significant environmental risks to both human health and the economy if it is not properly managed. As a result, scientists and decision-makers are paying increasing attention to developing a sustainable, low-cost wastewater treatment technique. This work aims to investigate the adsorption of Methylene Blue (MB) dye present in water using biochar derived from sawdust modified by boiling in an ammonia solution (SDBA). The properties of SDBA were characterized by BET, SEM, XRD, BJH, FT-IR, DTA, EDX and TGA analyses. The presence of -OH and -NH groups in SDBA was confirmed by FTIR, which proved that the NH4OH treatment of biochar successfully added nitrogen groups on its surface. The influence of pH (2 to 12), MB dye initial concentration (20 to 120 mg/L), adsorbent dosage (0.5 to 4.0 g/L) and contact time (0 to 180 min) on the adsorption process has been investigated. The adsorption of MB dye is more favorable at basic pH, with optimum adsorption at pH 8. Using a starting concentration of 20 mg/L of MB dye and a 4.0 g/L SDBA dose, the maximum percent clearance of MB dye was 99.94%. Experimental results were fitted to the Freundlich (FIM), Tempkin (TIM) and Langmuir (LIM) isotherm models (IMs). The FIM fitted the equilibrium data well, with a 643.74 mg/g Qm. Various error function models were used to test the data obtained from IMs. According to Error Function results, experimental data showed that it fits better for LIM and FIM. Kinetic studies indicated that the MB dye adsorption procedure followed pseudo-second-order (PSOM) kinetics based on film diffusion (FDM), pseudo-first-order (PFOM) and intra-particle diffusion models (IPDM). MB dye sorption on the SDBA involved electrostatic interaction, surface participation, hydrogen bond and π-π interactions. The adsorption mechanism of MB dye by SDBA was proposed as physical adsorption via the electrostatic attraction process. SDBA is an effective adsorbent in removing MB dye from water. Six adsorption-desorption cycles of the MB dye were run through the regeneration of SDBA with only a minimal amount of adsorption capacity loss, demonstrating the reusability of manufactured SDBA.


An intricate relationship between executive function and second-language ability in a cohort of Uyghur-Chinese bilingual children.
The relationship between executive function and second-language ability remains contentious in bilingual children; thus, the current study focused on this issue. In total, 371 Uyghur-Chinese bilingual children ranging from 3 to 6 years old were assessed by a battery of tasks measuring language ability (expressive vocabulary tests, receptive vocabulary tests, and phonological awareness of both their first-language and second-language) and executive function (working memory, inhibition, and switching). Our results indicated that age is a crucial moderator of the relationship between second-language ability and executive function. Specifically, executive function unilaterally predicted second-language ability in children who were 3-4 years old, whereas second-language ability and executive function bilaterally influenced each other in children who were 4-5 and 5-6 years old. These findings suggest that executive function and second-language ability have an intertwined and causal relationship among preschool children during development. The theoretical implications of these findings are discussed. RESEARCH HIGHLIGHTS: A counterfactual model showed causality between executive function and second language ability in Uyghur-Chinese bilingual children. Executive function unilaterally predicted second language ability in the 3- to 4-year-old age group. Executive function and second language ability bilaterally influenced each other in the 4- to 5 and 5- to 6-year-old age groups.


[Prenatal diagnosis and genetic analysis of a special case with complex structural rearrangements of chromosome 8].
To present on a prenatally diagnosed case with complex structural rearrangements of chromosome 8. Chromosome karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) were carried out for a fetus with increased nuchal thickness. The karyotype of the amniotic fluid sample showed extra materials on 8p. FISH revealed a centromeric signal at the terminal of 8p with absence of telomeric signal. CMA revealed partial deletion of 8p23.3 [(208049_2256732)×1], partial duplication of 8p23.3p23.2 [(2259519_3016818)×3], and partial duplication of 8q [8q11.1q12.2(45951900_60989083)×3]. The complex structural rearrangements of chromosome 8 in this case has differed from the commonly seen inv dup del(8p).


Effects of microclimate and human parameters on outdoor thermal sensation in the high-density tropical context of Dhaka.
A thermal comfort questionnaire survey was carried out in the high-density, tropical city Dhaka. Comfort responses from over 1300 subjects were collected at six different sites, alongside meteorological parameters. The effect of personal and psychological parameters was examined in order to develop predictive models. Personal parameters included gender, age, activity, profession-type (indoor or outdoor-based), exposure to air-conditioned space and sweat-levels. Psychological parameters, such as 'the reason for visiting the place' and 'next destination is air-conditioned', had statistically significant effects on thermal sensation. Other parameters, such as 'body type', 'body exposure to sun', 'time living in Dhaka', 'travelling in last_30 min', and 'hot food' did not have any significant impact. Respondents' humidity, wind speed and solar radiation sensation had profound impacts and people were found willing to adjust to the thermal situations with adaptive behaviour. Based on actual sensation votes from the survey, empirical models are developed to predict outdoor thermal sensation in the case study areas. Ordinal linear regression techniques are applied for predicting thermal sensation by considering meteorological and personal conditions of the field survey. The inclusion of personal and weather opinion factors produced an improvement in models based on meteorological factors. The models were compared with the actual thermal sensation using the cross-tabulation technique. The predictivity of the three models (meteorological, thermos-physiological and combined parameter) as expressed by the gamma coefficient were 0.575, 0.636 and 0.727, respectively. In all three models, better predictability was observed in the 'Slightly Warm' (71% in meteorological model) and 'Hot' (64.9% in combined parameter model) categories-the most important ones in a hot-humid climate.


[18F]FAPI PET/CT in the evaluation of focal liver lesions with [18F]FDG non-avidity.
This prospective study was aimed to investigate the potential utility of [18F]fibroblast activation protein inhibitor (FAPI) PET/CT for evaluating focal liver lesions (FLLs) with [18F]FDG non-avidity. From January 2021 to March 2022, this prospective study included 80 FLLs that were not avid on [18F]FDG PET/CT from 37 patients, then underwent [18F]FAPI PET/CT. All patients with FLL(s) with biopsy-proof or follow-up confirmation were categorized into four subgroups (20 hepatocellular carcinomas [HCCs]/5 non-HCC malignancies/4 inflammatory FLLs/8 benign noninflammatory FLLs). The diagnostic value of [18F]FAPI for detecting liver malignancy was determined by visual evaluation. Differences in the maximum standardized uptake value (SUVmax) and lesion-to-background ratio (LBR) obtained from [18F]FAPI PET/CT among the four subgroups were analyzed by semiquantitative analysis. Among the thirty-seven enrolled participants (34 males; median age 57 years, range 48-67 years), on visual evaluation, the sensitivity, specificity, and accuracy of [18F]FAPI PET for detecting liver malignancy in the patient-based analysis were 96.0% (24/25), 58.3% (7/12), and 83.8% (31/37), respectively. On semiquantitative analysis, the SUVmax and LBR of [18F]FAPI PET in liver malignancy (33 HCC lesions; 19 non-HCC malignant lesions) were significantly higher than those in 11 benign noninflammatory FLLs [HCC: SUVmax: 6.4 vs. 4.5, P = 0.017; LBR: 5.1 vs. 1.5, P = 0.003; non-HCC: SUVmax: 5.5 vs. 4.5, P = 0.008; LBR: 4.4 vs. 1.5, P = 0.042]. Notably, there was no significant difference in the SUVmax of [18F]FAPI PET between 33 HCC lesions and 17 inflammatory FLLs (6.4 vs. 8.2, P = 0.37), but the LBR of [18F]FAPI PET in HCC were significantly lower than that in inflammatory FLLs (5.1 vs. 9.1, P = 0.003). [18F]FAPI PET/CT shows high sensitivity in detecting HCC and non-HCC malignancy with [18F]FDG non-avidity. [18F]FAPI might be a promising radiopharmaceutical for the differential diagnosis of benign noninflammatory FLLs and liver malignancy with [18F]FDG non-avidity.


Influence of anthropogenic activities on the spatial distribution of heterotrophic, indicator and pathogenic bacteria around Andaman and Nicobar Islands.
To examine the influence of anthropogenic activities on the marine ecosystem around the Andaman and Nicobar Islands, a study was conducted to investigate the abundance of heterotrophic, indicator and pathogenic bacteria during the monsoon season. We noticed the higher abundance of heterotrophic, indicator (Escherichia coli and Enterococcus faecalis) and pathogenic bacterial counts (Aeromonas hydrophila, Enterobacter aerogens, Escherichia coli strain O157:H7, Shigella dysenteriae and Vibrio parahaemolyticus) near the 10o channel, which is the principal route to reach Andaman mainland. Most of the stations are offshore (chosen to cover shipping tracks) at a distance ranging from 41 to 266 km from the coast. The nearest station to the coast was at 21 km away. Apart from those mentioned above, several other pathogenic bacteria were also detected such as Klebsiella pneumoniae, Salmonella paratyphi, Vibrio cholera and Vibrio vulnificus but they are sparsely detected at few stations only.


Fluorinase mediated chemoenzymatic synthesis of [(18)F]-fluoroacetate.
A novel preparation of sodium [(18)F]-fluoroacetate is described where 5'-[(18)F]-fluoro-5'-deoxyadenosine is generated by a fluorinase catalysed reaction of S-adenosyl-l-methionine (SAM) with no carrier added [(18)F]-fluoride and then oxidation to [(18)F]-fluoroacetate by a Kuhn-Roth oxidative degradation.


Physiological interpretation of Doppler shift waveforms: the femorodistal segment in combined disease.
A new method is presented for assessing the femorodistal segment in multisegmental arterial disease, using the Laplace transform technique of Doppler waveform analysis. Blood velocity/time waveforms were obtained at femoral and ankle levels in three groups of limbs--50 without arterial disease, 12 with isolated aortoiliac stenoses, and 32 with femoropopliteal occlusions, with and without proximal disease. The waveforms were analysed for Laplace transform and pulsatility index values. The omega 0 coefficients of the Laplace transform analysis at femoral and ankle levels were compared in each subject, as the omega 0 gradient (femoral/ankle omega 0): and pulsatility index damping factor (femoral/ankle P1) was also calculated. The omega 0 gradient was shown to detect femoropopliteal occlusion in the presence of multisegmental arterial disease and to give some indication of its haemodynamic significance. The diagnostic accuracy of the omega 0 gradient was superior to that of pulsatility index damping factor. When combined with its existing ability to detect aortoiliac stenosis, this new application of the Laplace transform method offers the possibility both of a system for complete localisation of significant arterial lesions, and potential for follow-up of vascular surgical procedures in the lower limb, from two simple Doppler recordings.


Refusal to eat, capacity, and ethics in stroke patients: a report of 3 cases.
Patients have the right to make decisions about their health care, including refusing a particular treatment or a life-sustaining activity such as eating. However, patients must be able to understand and appreciate the consequences of their actions. Brain injury caused by a stroke has the potential to affect people's capacity to understand and appreciate their particular medical condition or its consequences. The purpose of this report was to describe the unique medical and ethical challenges presented by 3 rehabilitating stroke patients who refused to eat. Two patients had left-hemisphere strokes, with resultant aphasia. The third patient had a right-brain stroke and was able to state his position verbally but denied many of the consequences of his stroke. The refusal to eat and, therefore, possible impending death forced the attending physiatrist and rehabilitation team members to reevaluate the issue of capacity in the stroke patient. None of the patients died, and oral intake varied from fair to poor. This report elucidates the particular diagnostic, management, legal, and ethical issues surrounding the difficult but likely not uncommon issue of refusal to eat in stroke patients. It highlights the concept of capacity, which guides clinical decision making in such patients, and suggests specific clinical courses of action to take.


Effect of COVID-19 on BPSD severity and caregiver distress: Trend data from national dementia-specific behavior support programs in Australia.
Globally, Coronavirus disease 2019 (COVID-19) caused a significant disruption to the physical and mental well-being of all individuals, including those living with dementia. Social restrictions and lockdown measures due to COVID-19 have worsened the feelings of loneliness and behaviours and psychological symptoms of dementia (BPSD). National BPSD support programs in Australia are offered by Dementia Support Australia (DSA) through the Dementia Behavior Management Advisory Service (DBMAS) and the Severe Behavior Response Teams (SBRT). This study aims to investigate the impact of COVID-19 on BPSD severity and related caregiver distress among referrals to DSA programs. A retrospective comparative analysis was conducted on the intake data of referrals to DSA between two periods: Pre-COVID-19 Period (January 2018-Decmeber 2019) and COVID-19 Period (January 2020-July 2021). Referrals were compared on demographic characteristics (e.g., age), and BPSD severity (i.e., neuropsychiatric symptoms such as agitation) and caregiver distress as measured by the Neuropsychiatric Inventory (NPI). NPI scores were compared on a month-to-month basis between the specified periods. Across the two periods, there were a total of 23,180 referrals eligible for the analysis. While no differences were noted in age, sex, or dementia subtype, there were elevated levels of NPI severity and caregiver distress scores during COVID-19 Period compared to the Pre-COVID-19 Period. The month-to-month trends of these differences (Figure 1, Figure 2) reflect the timing of outbreaks across Australia. Specifically, there were no significant differences at the start of 2020 prior to the declaration of the pandemic, with an initial rise in NPI severity and distress through April after initial measures were implemented nationally in March 2020. These levels of severity and distress continued to rise through the remainder of 2020, alongside outbreaks in specific regions within Australia, such as Victoria (June-October), and New South Wales (December/January). NPI severity and caregiver distress then began to return to Pre-COVID levels from February-May before elevating again with the outbreak of the Delta variant in Australia. COVID-19 has a significant impact on the severity levels of BPSD and related caregiver distress.


Multifocal segmental hyperthermic and hyperhidrotic naevus flammeus: a peculiar variant of eccrine angiomatous hamartoma?
Eccrine angiomatous hamartoma (EAH) is a lesion that tends to appear as a plaque or nodular single lesion that may be painful and/or hyperhidrotic. Histopathologically, EAH is manifested by lobules of mature eccrine glands and ducts closely associated with well-differentiated, thin-walled blood vessels. Rarely, EAH can present as a macular hyperhidrotic lesion following a segmental distribution.


Learning capacities during sleep: a preliminary study in a group of schizophrenic inpatients.
The relationship between sleep and learning processes is analysed in a sample of schizophrenic patients, starting from more recent hypotheses about the function of REM sleep in learning and memory processes. This is done by means of two experiments: in the first AA. evaluate the possibility to elicit a simple motor conditional reflex acquired during daytime in different sleep stages. With the second experiment daytime learning performances are evaluated with and without a reinforcement administered during REM sleep. Results for the first experiment underline a qualitative difference between REM and nREM sleep in a reflexological perspective. In nREM sleep the conditional response is better maintained than in REM sleep. The second experiment confirms the possibility to improve daytime learning performances after an additional presentation of learning material in REM. The joint study of sleep abnormalities and learning and cognitive impairment in schizophrenic patients is finally suggested.


Temporal variation in habitat structure and shrubsteppe bird dynamics.
We investigate the role of temporal variation in habitat physiognomy in influencing the dynamics of shrubsteppe bird populations and communities. During a 3-y (1977-1979) study of 14 sites in the northern Great Basin of North America, annual precipitation varied substantially, with one of the driest years on record followed by one of the wettest. This resulted in significant physiognomic variation (increasing height and coverage of vegetation, decreasing horizontal patchiness), mediated largely by changes in the annual elements of the flora, particularly forbs and grasses. Shrub species coverage values, on the other hand, demonstrated no statistically detectable year-to-year changes, nor were they correlated with any physiognomic variation. Despite large scale physiognomic changes, no bird species' abundance varied in a statistically significant fashion; neither could variation in bird abundances be correlated with variation in either physiognomy or shrub species coverages.Multivariate analyses revealed essentially the same patterns as the univariate analyses: substantial changes in physiognomy, few changes in shrub species coverages or bird species abundances, and little correlation of temporal variation among the three data sets. Calculation of the Euclidean distances that sites "moved" in multivariate physiognomic, bird species, or shrub species hyperspaces yields synthetic gradients of annual "turnover" of sites with respect to those data sets. Sites identified as demonstrating high physiognomic turnover were characterized by high coverage of grass and forbs, while low turnover sites had greater coverage of shrubs and higher shrub species diversity. Relatively high bird turnover sites had greater numbers of Western Meadowlarks and Black-throated Sparrows, while more stable sites had high numbers of Brewer's Sparrows. Physiognomically, high bird turnover sites were grassier and had greater total vegetation coverage, while low bird turnover sites had more bare ground and higher horizontal patchiness. A site's position on the avifauna turnover axis, however, was uncorrelated with its position on the physiognomic turnover axis. Shrub species showed virtually no annual turnover.Reanalysis of a previous Principal Components Analysis (PCA) of these same data sets that was applied without regard to year of sampling revealed that the first physiognomic component (41% of the total physiognomic variability) did in fact have a strong temporal element, and that this element was consistent with the changes in univariate characters noted above. No other physiognomic component could be associated with annual variation, nor could any components of parallel bird abundance or shrub species coverage PCA's.Regional patterns indicate that sites tended to be very consistent from year to year in their relationships to one another as defined by their relative locations in either physiognomic or shrub species hyperspaces, but varied independently of one another with respect to their bird species abundances and composition. Examination of the temporal consistency of site relationships between "bird space" and "vegetation space" reveals that bird communities are to a large degree independent of a site's physiognomic position, but instead are strongly associated with its position in shrub species hyperspace.The overall patterns that emerge from these analyses are consistent with the so-called "checkerboard effect" that results from the apparently random annual redistribution of individual birds, and leads to the conclusion that populations of shrubsteppe birds are not existing at maximum density or "carrying capacity." Such observations are consistent with contentions that these populations lack close biological coupling with coexisting species and that interactions among these species (e.g. competition) are likely to play little if any role in the organization of their communities.


1α,25-dihydroxyvitamin D inhibits de novo fatty acid synthesis and lipid accumulation in metastatic breast cancer cells through down-regulation of pyruvate carboxylase.
Both increased de novo fatty acid synthesis and higher neutral lipid accumulation are a common phenotype observed in aggressive breast cancer cells, making lipid metabolism a promising target for breast cancer prevention. In the present studies, we demonstrate a novel effect of the active metabolite of vitamin D, 1α,25-dihydroxyvitamin D (1,25(OH)₂D) on lipid metabolism in malignant breast epithelial cells. Treatment of MCF10CA1a breast epithelial cells with 1,25(OH)₂D (10 nM) for 5 and 7 days decreased the level of triacylglycerol, the most abundant form of neutral lipids, by 20%(±3.9) and 50%(±5.9), respectively. In addition, 1,25(OH)₂D treatment for 5 days decreased palmitate synthesis from glucose, the major fatty acid synthesized de novo (48%±5.5 relative to vehicle). We have further identified the anaplerotic enzyme pyruvate carboxylase (PC) as a target of 1,25(OH)₂D-mediated regulation and hypothesized that 1,25(OH)₂D regulates breast cancer cell lipid metabolism through inhibition of PC. PC mRNA expression was down-regulated with 1,25(OH)₂D treatment at 2 (73%±6 relative to vehicle) and 5 (56%±8 relative to vehicle) days. Decrease in mRNA abundance corresponded with a decrease in PC protein expression at 5 days of treatment (54%±12 relative to vehicle). Constitutive overexpression of PC in MCF10CA1a cells using a pCMV6-PC plasmid inhibited the effect of 1,25(OH)₂D on both TAG accumulation and de novo palmitate synthesis from glucose. Together, these studies demonstrate a novel mechanism through which 1,25(OH)₂D regulates lipid metabolism in malignant breast epithelial cells.


Three-Enzyme Phosphorylase Cascade for Integrated Production of Short-Chain Cellodextrins.
Cellodextrins are linear β-1,4-gluco-oligosaccharides that are soluble in water up to a degree of polymerization (DP) of ≈6. Soluble cellodextrins have promising applications as nutritional ingredients. A DP-controlled, bottom-up synthesis from expedient substrates is desired for their bulk production. Here, a three-enzyme glycoside phosphorylase cascade is developed for the conversion of sucrose and glucose into short-chain (soluble) cellodextrins (DP range 3-6). The cascade reaction involves iterative β-1,4-glucosylation of glucose from α-glucose 1-phosphate (αGlc1-P) donor that is formed in situ from sucrose and phosphate. With final concentration and yield of the soluble cellodextrins set as targets for biocatalytic synthesis, three major factors of reaction efficiency are identified and partly optimized: the ratio of enzyme activity, the ratio of sucrose and glucose, and the phosphate concentration used. The efficient use of the phosphate/αGlc1-P shuttle for cellodextrin production is demonstrated and the soluble product at 40 g L-1 is obtained under near-complete utilization of the donor substrate offered (88 mol% from 200 mm sucrose). The productivity is 16 g (L h)-1 . Through a simple two-step route, the soluble cellodextrins are recovered from the reaction mixture in ≥95% purity and ≈92% yield. Overall, this study provides the basis for their integrated production.


Coordinating care: the community health record.
Stepping boldly into the future, the PeaceHealth network connects all the communities it serves in Oregon, Washington and Alaska with a fully digital online electronic patient record.


Proteomic Profiling of Paracoccidioides brasiliensis in Response to Phenacylideneoxindol Derivative: Unveiling Molecular Targets and Pathways.
The treatment of paracoccidioidomycosis (PCM) is a challenge, and the discovery of new antifungal compounds is crucial. The phenacylideneoxindoles exhibited promising antifungal activity against Paracoccidioides spp., but their mode of action remains unknown. Through proteomic analysis, we investigated the effects of (E)-3-(2-oxo-2-phenylethylidene)indolin-2-one on P. brasiliensis. In addition, we investigated the metabolic alterations of P. brasiliensis in response to the compound. Furthermore, the effects of the compound on the membrane, ethanol production, and reactive oxygen species (ROS) production were verified. We identified differentially regulated proteins that revealed significant metabolic reorganization, including an increase in ethanol production, suggesting the activation of alcoholic fermentation and alterations in the rigidity of fungal cell membrane with an increase of the ergosterol content and formation of ROS. These findings enhance our understanding of the mode of action and response of P. brasiliensis to the investigated promising antifungal compound, emphasizing its potential as a candidate for the treatment of PCM.


Mnemonic and neuropathological effects of occluding the posterior cerebral artery in Macaca mulatta.
To investigate experimentally the mnemonic and neuropathological effects of blockage of the posterior cerebral arteries (PCA), a cerebrovascular accident that can lead to global anterograde amnesia in humans, we permanently occluded these arteries bilaterally in six monkeys and then evaluated their performance on a visual recognition task, after which we assessed the extent of their ischemic infarcts. The latter showed substantial individual variation, ranging from almost no damage in one case to massive unilateral injury of both the ventromedial o occipitotemporal cortex and hippocampal formation in another. In the four remaining cases, however, the infarcts fell within a narrow range, being confined almost entirely to the hippocampal formation and parahippocampal gyrus, and then only to restricted portions of these structures, unilaterally in one case, and bilaterally in the three others. Performance on the recognition task was related to the presence and bilaterality of the hippocampal injury. Thus, the case without any hippocampal damage performed at a rate equal to that of normal controls; the case with unilateral hippocampal damage was mildly impaired; and the three cases with bilateral infarctions, involving between 20 and 55% of the hippocampal formation, showed substantial impairment, with scores averaging 20% below those of normal controls. The only subfields of the hippocampus damaged in common in these cases were CA1 and CA2. Paradoxically, the memory loss found in these three animals with only partial bilateral hippocampal damage was significantly greater than that found in animals with total bilateral ablation of the hippocampal formation, whose scores averaged only 10% below those of normal controls. Possible explanations for this extremely puzzling outcome are proposed.


Factors influencing the outcome of treatment in patients referred to a temporomandibular joint clinic.
A retrospective review was performed in 350 consecutive patients referred for functional disturbances of the masticatory system. Two and one-half years after the first visit, the records of all patients were examined for clinical findings, diagnosis, and observations related to treatment. Number of visits, length of the treatment period, and evaluation of signs and symptoms at last visit (factors which showed a wide variation) were analyzed for correlation with signs and symptoms at first visit and some background factors. A total of 50 variables were considered. The number of visits and length of treatment indicated the severity of signs and symptoms at the first visit. Multiple regression analysis showed that tenderness on muscle palpation at first visit was the most important of the independent variables. About one-fourth of the variance of number of visits and length of treatment could be explained (R2 = 0.28 and 0.27, respectively), while the corresponding value for the variance of the evaluation at last visit was 15% (R2 = 0.15). The results tend to emphasize the heterogeneity of patients with functional disturbances of the masticatory system and the difficulty of prognosticating the outcome of treatment in such patients, even when attempts are made to divide them into subgroups.


Modelling salinity inhibition effects during biodegradation of perchlorate.
To determine the mathematical kinetic rates and mechanisms of acclimated perchlorate (ClO)-reducing microbial cultures by incorporating a term to relate the inhibitory effect of high salinity during biological reduction of concentrated perchlorate solutions. Salt toxicity associated with the biodegradation of concentrated perchlorate (200, 500, 1100, 1700 and 2400 mg l(-1) as ClO) was investigated using two microbial cultures isolated from a domestic wastewater treatment plant [return activated sludge (RAS) and anaerobic digester sludge (ADS)]. Experiments were performed in wastewaters containing various sodium chloride concentrations, ranging from 0% to 4.0% (w/v) NaCl (ionic strength: 0.14-0.82 mol l(-1), total dissolved solids: 5.3-42.6 g l(-1)) at near-neutral values of pH (6.7-7.8). Perchlorate biodegradation was stimulated through stepwise acclimation to high salinity. The ADS culture was capable of reducing perchlorate at salinities up to 4% NaCl, while the RAS culture exhibited complete inhibition of perchlorate degradation at 4% NaCl, probably resulting from either a toxic effect or enzyme inactivation of the perchlorate-reducing microbes. Further, a kinetic growth model was developed based on experimental data in order to express an inhibition function to relate specific growth rate and salinity. Biological reduction of concentrated perchlorate wastewaters using either acclimated RAS or ADS cultures is feasible up to 3% or 4% NaCl, respectively. In addition, the kinetic model including a salinity inhibition term should be effective in many practical applications such as improving reactor design and management, furthering the understanding of high salinity inhibition, and enhancing bioremediation under high salinity loading conditions. Applications of these findings in water treatment practice where ion exchange or membrane technologies are used to remove perchlorate from water can have the potential to increase the overall attractiveness of these processes by eliminating the need to dispose of a concentrated perchlorate solution.


Concerted epithelial and stromal changes during progression of Barrett's Esophagus to invasive adenocarcinoma exposed by multi-scale, multi-omics analysis.
Esophageal adenocarcinoma arises from Barrett's esophagus, a precancerous metaplastic replacement of squamous by columnar epithelium in response to chronic inflammation. Multi-omics profiling, integrating single-cell transcriptomics, extracellular matrix proteomics, tissue-mechanics and spatial proteomics of 64 samples from 12 patients' paths of progression from squamous epithelium through metaplasia, dysplasia to adenocarcinoma, revealed shared and patient-specific progression characteristics. The classic metaplastic replacement of epithelial cells was paralleled by metaplastic changes in stromal cells, ECM and tissue stiffness. Strikingly, this change in tissue state at metaplasia was already accompanied by appearance of fibroblasts with characteristics of carcinoma-associated fibroblasts and of an NK cell-associated immunosuppressive microenvironment. Thus, Barrett's esophagus progresses as a coordinated multi-component system, supporting treatment paradigms that go beyond targeting cancerous cells to incorporating stromal reprogramming.


[Study of the hypothalamo-pituitary adrenal function in 16 patients after surgery for pituitary tumor (author's transl)].
The hypothalamic pituitary adrenocortical function has been studied in 16 patients operated from pituitary tumors (13 adenomas; 3 craniopharyngiomas). Comparisons have been made between corticotropin and cortisol response to lysine vasopressin, insulin induced-hypoglycemia and metyrapone IV and per os. Among these different stimulating tests, insulin induced hypoglycemia and metyrapone per os seem to give the more accurate informations metyrapone per os being more convenient because harmless. Three different groups of patients have been distinguished : one without adrenocortical deficiency; one with a complete deficiency and a third group with a partial deficiency. Correlations have been studied between the degree of the adrenocortical deficiency, the volume of the tumor and the presence of the absence of other anterior pituitary dysfunctions.


To die or not to die, does it change the function? Behavior of transgenic mice reveals a role for developmental cell death.
In humans, perturbations in the developmental neuronal death leading to an excess of neurons could be associated with developmental neuropsychiatric disorders. Hu-bcl-2 transgenic mice appear to be a valuable tool to study the functional role of developmental programmed cell death. Indeed, the over-expression of the anti-apoptotic gene bcl-2 decreases developmental neuronal death and Hu-bcl-2 mice present supernumerary neurons in several brain regions. A detailed behavioral analysis of these mice revealed selective deficits. Hu-bcl-2 mice have normal vision, general activity and motor skills. Only the most complex behavior like anxiety and learning abilities are impaired in these mice.


Probabilistic DHP adaptive critic for nonlinear stochastic control systems.
Following the recently developed algorithms for fully probabilistic control design for general dynamic stochastic systems (Herzallah & Káarnáy, 2011; Kárný, 1996), this paper presents the solution to the probabilistic dual heuristic programming (DHP) adaptive critic method (Herzallah & Káarnáy, 2011) and randomized control algorithm for stochastic nonlinear dynamical systems. The purpose of the randomized control input design is to make the joint probability density function of the closed loop system as close as possible to a predetermined ideal joint probability density function. This paper completes the previous work (Herzallah & Káarnáy, 2011; Kárný, 1996) by formulating and solving the fully probabilistic control design problem on the more general case of nonlinear stochastic discrete time systems. A simulated example is used to demonstrate the use of the algorithm and encouraging results have been obtained.


The role of chemical fractionation in risk assessment of toxic metals: a review.
The identification of highly toxic metals like Cd, Ni, Pb, Cr, Co or Cu in ambient particulate matter (PM) has garnered a lot of interest recently. Exposure to toxic metals, including carcinogenic ones, at levels above recommended limits, can significantly affect human health. Prolonged exposure to even trace amounts of toxic or essential metals can also have negative health impacts. In order to assess significant risks, it is crucial to govern the concentrations of bioavailable/bio-accessible metals that are available in PM. Estimating the total metal concentrations in PM is only an approximation of metal toxicity. This review provides an overview of various procedures for extracting soluble toxic metals from PM and the importance of chemical fractionation in risk assessment. It is observed that the environmental risk indices such as bioavailability index (BI), contamination factor (CF) and risk assessment code (RAC) are specifically influenced by the concentration of these metals in a particular fraction. Additionally, there is compelling evidence that health risks assessed using total metal concentrations may be overestimated, therefore, the metal toxicity assessment is more accurate and more sensitive to the concentration of the bioavailable/bio-accessible fraction than the total metal concentrations. Hence, chemical fractionation of toxic metals can serve as an effective tool for developing environmental protection laws and improving air quality monitoring programs for public health.


Analysis of FMRFamide-like peptide (FLP) diversity in phylum Nematoda.
This study reports a series of systematic BLAST searches of nematode ESTs on the Genbank database, using search strings derived from known nematode FLPs (those encoded by Caenorhabditis elegans flp genes as well as those isolated from other nematodes including Ascaris suum), as well as query sequences representative of theoretical FLPs. Over 1000 putative FLP-encoding ESTs were identified from multiple nematode species. A total of 969 ESTs representing sequelogs of the 23 known C. elegans flp genes were identified in 32 species, from clades I, III, IV and V. Numerical analysis of EST numbers suggests that flp-1, flp-11 and flp-14 are amongst the most highly expressed flp genes. Speculative BLAST searches were performed using theoretical FLP C-termini as queries, in an attempt to identify putative novel FLP sequences in the EST database. These searches yielded eight multi-species sequelogs encoding FLPs with novel signatures that are believed to identify distinct flp genes. These novel genes encode 25 distinct previously unidentified FLPs, and raise the current total of known nematode flp genes to 31. Additionally, software-based analyses of the presence of signal peptides were performed, with signal peptides being identified on at least one member of each group of flp ESTs, further confirming their status as secreted peptides. The data reveal that nematode FLPs encompass the most complex neuropeptide family known within the metazoa. Moreover, individual FLPs and FLP motifs are highly conserved across the nematodes with little evidence for inter-clade or inter-lifestyle variation, supporting their fundamental role in free-living and parasitic species.


Porcine vocal fold elasticity evaluation using Brillouin spectroscopy.
The vocal folds are critically important structures within the larynx which serve the essential functions of supporting the airway, preventing aspiration, and phonation. The vocal fold mucosa has a unique multilayered architecture whose layers have discrete viscoelastic properties facilitating sound production. Perturbations in these properties lead to voice loss. Currently, vocal fold pliability is inferred clinically using laryngeal videostroboscopy and no tools are available for in vivo objective assessment. The main objective of the present study is to evaluate viability of Brillouin microspectroscopy for differentiating vocal folds' mechanical properties against surrounding tissues. We used Brillouin microspectroscopy as an emerging optical imaging modality capable of providing information about local viscoelastic properties of tissues in noninvasive and remote manner. Brillouin measurements of the porcine larynx vocal folds were performed. Elasticity-driven Brillouin spectral shifts were recorded and analyzed. Elastic properties, as assessed by Brillouin spectroscopy, strongly correlate with those acquired using classical elasticity measurements. These results demonstrate the feasibility of Brillouin spectroscopy for vocal fold imaging. With more extensive research, this technique may provide noninvasive objective assessment of vocal fold mucosal pliability toward objective diagnoses and more targeted treatments.


Energy saving strategies of honeybees in dipping nectar.
The honeybee's drinking process has generally been simplified because of its high speed and small scale. In this study, we clearly observed the drinking cycle of the Italian honeybee using a specially designed high-speed camera system. We analysed the pattern of glossal hair erection and the movement kinematics of the protracting tongue (glossa). Results showed that the honeybee used two special protraction strategies to save energy. First, the glossal hairs remain adpressed until the end of the protraction, which indicates that the hydraulic resistance is reduced to less than 1/3 of that in the case if the hairs remain erect. Second, the glossa protracts with a specific velocity profile and we quantitatively demonstrated that this moving strategy helps reduce the total energy needed for protraction compared with the typical form of protraction with constant acceleration and deceleration. These findings suggest effective methods to optimise the control policies employed by next-generation microfluidic pumps.


Weather system scale variation in radon-222 concentration of indoor air.
Radon-222 concentration measurements using the charcoal adsorption technique were made continuously over periods of up to 2 years in three houses, two rural and one urban. The measurement records for all houses show a variation in radon concentration on a seasonal scale on which is superimposed shorter periods of relatively large magnitude fluctuations on a scale longer than the typically 4-5-day measurement duration. Regression analysis using meteorological data from a weather station remote from the houses shows that much of the variation in both monthly and 3-day mean radon concentration can be explained by regional scale external temperature variation. Wind speed, rainfall and barometric pressure apparently influence the 3-day values to a much smaller degree. Differences in the radon levels between the houses reflect the different geological radon potential of the two areas, with notably higher levels over limestone than glacial till and sandstone. Other differences within and between houses reflect house construction and occupancy factors.


First example of the correlated calculation of the one-bond tellurium-carbon spin-spin coupling constants: Relativistic effects, vibrational corrections, and solvent effects.
This work reports on the comprehensive calculation of the NMR one-bond spin-spin coupling constants (SSCCs) involving carbon and tellurium, (1) J((125) Te,(13) C), in four representative compounds: Te(CH3 )2 , Te(CF3 )2 , Te(CCH)2 , and tellurophene. A high-level computational treatment of (1) J((125) Te,(13) C) included calculations at the SOPPA level taking into account relativistic effects evaluated at the 4-component RPA and DFT levels of theory, vibrational corrections, and solvent effects. The consistency of different computational approaches including the level of theory of the geometry optimization of tellurium-containing compounds, basis sets, and methods used for obtainig spin-spin coupling values have also been discussed in view of reproducing the experimental values of the tellurium-carbon SSCCs. Relativistic corrections were found to play a major role in the calculation of (1) J((125) Te,(13) C) reaching as much as almost 50% of the total value of (1) J((125) Te,(13) C) while relativistic geometrical effects are of minor importance. The vibrational and solvent corrections account for accordingly about 3-6% and 0-4% of the total value. It is shown that taking into account relativistic corrections, vibrational corrections and solvent effects at the DFT level essentially improves the agreement of the non-relativistic theoretical SOPPA results with experiment. © 2016 Wiley Periodicals, Inc.


A line-scanning semi-confocal multi-photon fluorescence microscope with a simultaneous broadband spectral acquisition and its application to the study of the thylakoid membrane of a cyanobacterium Anabaena PCC7120.
We describe the construction and characterization of a laser-line-scanning microscope capable of detection of broad fluorescence spectra with a resolution of 1 nm. A near-infrared femtosecond pulse train at 800 nm was illuminated on a line (one lateral axis, denoted as X axis) in a specimen by a resonant scanning mirror oscillating at 7.9 kHz, and total multi-photon-induced fluorescence from the linear region was focused on the slit of an imaging polychromator. An electron-multiplying CCD camera was used to resolve fluorescence of different colours at different horizontal pixels and fluorescence of different spatial positions in a specimen at different vertical pixels. Scanning on the other two axes (Y and Z) was achieved by a closed-loop controlled sample scanning stage and a piezo-driven objective actuator. The full widths at half maximum of the point-spread function of the system were estimated to be 0.39-0.40, 0.33 and 0.56-0.59 mum for the X (lateral axis along the line-scan), Y (the other lateral axis) and Z axes (the axial direction), respectively, at fluorescence wavelengths between 644 and 690 nm. A biological application of this microscope was demonstrated in a study of the sub-cellular fluorescence spectra of thylakoid membranes in a cyanobacterium, Anabaena PCC7120. It was found that the fluorescence intensity ratio between chlorophyll molecules mainly of photosystem II and phycobilin molecules of phycobilisome (chlorophyll/phycobilin), in the thylakoid membranes, became lower as one probed deeper inside the cells. This was attributable not to position dependence of re-absorption or scattering effects, but to an intrinsic change in the local physiological state of the thylakoid membrane, with the help of a transmission spectral measurement of sub-cellular domains. The efficiency of the new line-scanning spectromicroscope was estimated in comparison with our own point-by-point scanning spectromicroscope. Under typical conditions of observing cyanobacterial cells, the total exposure time became shorter by about 50 times for a constant excitation density. The improvement factor was proportional to the length of the line-scanned region, as expected.


Epidemiology of Heart Failure in Europe.
Heart failure (HF) is a major public health problem. For acute HF, therapeutic developments have been scarce in the past decades. In contrast to acute HF, chronic HF was the object of several successful controlled studies conducted in the past 30 years, which encouraged the use of drugs and devices able to improve the outcomes of ambulatory patients. For both patients with acute and chronic HF, observational research remains an important research tool to confirm the results of the controlled trials in the real world, to collect periodic reports, and to assess the quality-of-care indicators.


Deoxyspergualin directly suppresses antibody formation in vivo and in vitro.
The effect of deoxyspergualin (DSG, NKT-01) on humoral immunity was investigated both in vitro and in vivo. DSG inhibited the primary and secondary responses to T cell-dependent antigens and the response to T cell-independent antigens in thymic and athymic mice. However, natural antibodies in non-sensitized mice were affected less by the administration of DSG. The agent produced a dose-dependent inhibition of B cell proliferation and antibody production to lipopolysaccharide in vitro. Suppression of secondary antibody response was also shown, whenever antigen stimulation was not given, antibody production was not affected. These results suggest that DSG affects the proliferative stage of B lymphocytes in such a way as to inhibit their growth and antibody production.


Safety and tolerability of SARS-Cov-2 vaccination in patients with myasthenia gravis: A multicenter experience.
During the COVID-19 pandemic, myasthenia gravis (MG) patients have been identified as subjects at high risk of developing severe COVID-19, and thus were offered vaccination with priority. The lack of direct data on the safety and tolerability of SARS-CoV-2 vaccines in MG have contributed to vaccine hesitancy. To address this issue, the safety and tolerability of SARS-CoV-2 vaccines were assessed in a large cohort of MG patients from two referral centers. Patients with confirmed MG diagnosis, consecutively seen between October and December 2021 at two MG centers, were enrolled. Demographics, clinical characteristics, and information regarding SARS-CoV-2 infection/vaccination were extracted from medical reports and/or collected throughout telephonic or in-person interviews. Ninety-eight (94.2%) of 104 patients included were administered at least two vaccine doses 4 weeks before the interview or earlier, and among them, 63 of 98 (64.2%) have already received the "booster" dose. The most frequently used vaccines were BNT162b2-Pfizer-BioNTech and mRNA-1273-Moderna. Overall, only minor side effects were reported, most commonly local pain and fever. MG worsening after vaccination was observed in eight of 104 (7.7%) cases. The frequency of worsening among muscle-specific tyrosine kinase MG cases (3/9, 33.3%) was significantly higher compared to other serological subgroups. Spontaneous symptom regression was observed in six of eight cases. Twelve of 104 (11.5%) patients had SARS-CoV-2 infection, and none of the SARS-CoV-2-infected MG patients worsened after vaccination. Our data support the safety and tolerability of mRNA COVID-19 vaccines, which should be strongly recommended in MG patients, who could be at higher risk of complications if exposed to SARS-CoV-2 infection.


Assessment of silver nanoparticle-induced physiological and molecular changes in Arabidopsis thaliana.
In this study, the effect of silver nanoparticles and silver ions on Arabidopsis thaliana was investigated at physiological and molecular levels. The seedlings were grown in sublethal concentrations of silver nanoparticles and silver ions (0.2, 0.5, and 1 mg/L) in 1/4 Hoagland's medium for 14 days under submerged hydroponic conditions. Significantly higher reduction in the total chlorophyll and increase in anthocyanin content were observed after exposure to 0.5 and 1 mg/L silver nanoparticles as compared to similar concentrations of silver ions. Lipid peroxidation increased significantly after exposure to 0.2, 0.5, and 1 mg/L of silver nanoparticles and 0.5 and 1 mg/L of silver ions. Qualitative analysis with dichloro-dihydro-fluorescein diacetate and rhodamine 123 fluorescence showed a dose-dependent increase in reactive oxygen species production and changes in mitochondrial membrane potential in the roots of seedlings exposed to different concentrations of silver nanoparticles. Real-time PCR analysis showed significant upregulation in the expression of sulfur assimilation, glutathione biosynthesis, glutathione S-transferase, and glutathione reductase genes upon exposure to silver nanoparticles as compared with silver ions. Overall, based on the physiological and molecular level responses, it was observed that exposure to silver nanoparticles exerted more toxic response than silver ions in A. thaliana.


Deletion of SIRT3 inhibits osteoclastogenesis and alleviates aging or estrogen deficiency-induced bone loss in female mice.
SIRT3 has been speculated to affect osteoclast activity through its important roles in regulating mitochondrial function. It remains unclear whether SIRT3 affects osteoclast activity in female mice which is relevant to postmenopausal osteoporosis. We hypothesized that deletion of SIRT3 could modulate bone remodeling in female mice under physiological aging process or ovariectomy (OVX)-induced bone loss. We found that SIRT3 level was markedly increased in primary bone marrow-derived macrophages (BMMs) from both 26-month-old aged mice and OVX mice. Knockdown of SIRT3 in vitro inhibited osteoclast differentiation and mitochondrial biogenesis, and deletion of SIRT3 increased trabecular bone mass in female mice due to impaired osteoclastogenesis. The effect of SIRT3 on bone remodeling appears to be age-dependent as revealed by comparing the effect of SIRT3 deletion on 5-week-old, 3-month-old and 6-month-old female mice. Interestingly, Sirt3-/- mice were more resistant to bone loss following estrogen deficiency resulting from OVX. Our findings demonstrated that SIRT3 could play critical roles in bone remodeling and estrogen deficiency-induced bone loss in female mice, suggesting that SIRT3 and its downstream effectors might be potential novel therapeutic targets for the management of postmenopausal osteoporosis.


Targeted NF1 cancer therapeutics with multiple modes of action: small molecule hormone-like agents resembling the natural anticancer metabolite, 2-methoxyoestradiol.
Both the number and size of tumours in NF1 patients increase in response to the rise in steroid hormones seen at puberty and during pregnancy. The size of tumours decreases after delivery, suggesting that hormone-targeting therapy might provide a viable new NF1 treatment approach. Our earlier studies demonstrated that human NF1 tumour cell lines either went through apoptosis or ceased growth in the presence of 2-methoxyoestradiol (2ME2), a naturally occurring anticancer metabolite of 17-β estradiol. Previous reports of treatment with sulfamoylated steroidal and non-steroidal derivatives of 2ME2 showed promising reductions in tumour burden in hormone-responsive cancers other than NF1. Here we present the first studies indicating that 2ME2 derivatives could also provide an avenue for treating NF1, for which few treatment options are available. STX3451, (2-(3-Bromo-4,5-dimethoxybenzyl)-7-methoxy-6-sulfamoyloxy-1,2,3,4-tetrahydroisoquinoline), a non-steroidal sulphamate analogue of 2ME2, was tested in dose-dependent studies of malignant and benign NF1 human tumour cell lines and cell lines with variable controlled neurofibromin expression. The mechanisms of action of STX3451 were also analysed. We found that STX3451-induced apoptosis in human malignant peripheral nerve sheath tumour (MPNST) cell lines, even in the presence of elevated oestrogen and progesterone. It inhibits both PI3 kinase and mTOR signalling pathways. It disrupts actin- and microtubule-based cytoskeletal structures in cell lines derived from human MPNSTs and in cells derived from benign plexiform neurofibromas. STX3451 selectively kills MPNST-derived cells, but also halts growth of other tumour-derived NF1 cell lines. STX3451 provides a new approach for inducing cell death and lowering tumour burden in NF1 and other hormone-responsive cancers with limited treatment options.


Basilar migraine with severe EEG abnormalities.
We report a patient suffering from a series of attacks fulfilling the criteria of basilar migraine. During the attack there was no normal EEG background pattern and the EEG showed predominantly delta activity. After the attack the EEG returned to (nearly) normal in a very short time. Up to now no such EEG findings have been reported in basilar migraine.


Next-generation finely controlled graded porous antibacterial bioceramics for high-efficiency vascularization in orbital reconstruction.
Eyeball loss due to severe ocular trauma, intraocular malignancy or infection often requires surgical treatment called orbital implant reconstruction to rehabilitate the orbital volume and restore the aesthetic appearance. However, it remains a challenge to minimize the postoperative exposure and infection complications due to the inert nature of conventional orbital implants. Herein, we developed a novel Ca-Zn-silicate bioceramic implant with multi-functions to achieve the expected outcomes. The porous hardystonite (Ca2ZnSi2O7) scaffolds with triply periodic minimal surfaces (TPMS)-based pore architecture and graded pore size distribution from center to periphery (from 500 to 800 μm or vice versa) were fabricated through the digital light processing (DLP) technique, and the scaffolds with homogeneous pores (500 or 800 μm) were fabricated as control. The graded porous scaffolds exhibited a controlled bio-dissolving behavior and intermediate mechanical strength in comparison with the homogeneous counterparts, although all of porous implants presented significant antibacterial potential against S. aureus and E. coli. Meanwhile, the pore size-increasing scaffolds indicated more substantial cell adhesion, cell viability and angiogenesis-related gene expression in vitro. Furthermore, the gradually increasing pore feature exhibited a stronger blood vessel infiltrating potential in the dorsal muscle embedding model, and the spherical implants with such pore structure could achieve complete vascularization within 4 weeks in the eyeball enucleation rabbit models. Overall, our results suggested that the novel antibacterial hardystonite bioceramic with graded pore design has excellent potential as a next-generation orbital implant, and the pore topological features offer an opportunity for the improvement of biological performances in orbital reconstruction.


MicroRNA137-loaded lipid nanoparticles regulate synaptic proteins in the prefrontal cortex.
Genome-wide association studies indicate that allele variants in MIR137, the host gene of microRNA137 (miR137), confer an increased risk of schizophrenia (SCZ). Aberrant expression of miR137 and its targets, many of which regulate synaptic functioning, are also associated with an increased risk of SCZ. Thus, miR137 represents an attractive target aimed at correcting the molecular basis for synaptic dysfunction in individuals with high genetic risk for SCZ. Advancements in nanotechnology utilize lipid nanoparticles (LNPs) to transport and deliver therapeutic RNA. However, there remains a gap in using LNPs to regulate gene and protein expression in the brain. To study the delivery of nucleic acids by LNPs to the brain, we found that LNPs released miR137 cargo and inhibited target transcripts of interest in neuroblastoma cells. Biodistribution of LNPs loaded with firefly luciferase mRNA remained localized to the mouse prefrontal cortex (PFC) injection site without circulating to off-target organs. LNPs encapsulating Cre mRNA preferentially co-expressed in neuronal over microglial or astrocytic cells. Using quantitative proteomics, we found miR137 modulated glutamatergic synaptic protein networks that are commonly dysregulated in SCZ. These studies support engineering the next generation of brain-specific LNPs to deliver RNA therapeutics and improve symptoms of central nervous system disorders.


Liver fibrosis alters the molecular structures of hepatic glycogen.
Liver fibrosis (LF) leads to liver failure and short survival. Liver glycogen is a hyperbranched glucose polymer, comprising individual β particles, which can bind together to form aggregated α particles. Glycogen functionality depends on its molecular structure. This study compared the molecular structure of liver glycogen from both LF and healthy rats, and explored underlying mechanisms for observed differences. Glycogen from both groups contained α and β particles; the LF group contained a higher proportion of β particles, with the glycogen containing fewer long chains than seen in the control group. Both glycogen branching enzyme and glycogen phosphorylase showed a significant decrease of activity in the LF group. Transcriptomics and proteomics revealed a functional deficiency of mitochondria in the LF group, which may lead to changes in glycogen structure. These results provide for the first time an understanding of how liver fibrosis affects liver glycogen metabolism and glycogen structure. HYPOTHESIS: We hypothesized that the molecular structure of liver glycogen from a rat model of liver fibrosis would be altered compared to the control group.


Late Presentation of Giant Osteochondroma of Talar Neck in Elderly: A Case Report and Review of Literature.
Osteochondroma is a benign bone tumor of the bone primarily seen in younger age groups. However, late presentation of the same is a rare incidence, as the symptoms develop rapidly due to compression of nearby structures. We present a case of a 55-year-old male patient with a giant osteochondroma originating from the neck of the talus. The patient presented with a huge 100 × 70 × 50 mm swelling over the ankle. The patient underwent an excision of the swelling. Histopathological examination of the swelling confirmed the findings of an osteochondroma. The patient had an uneventful recovery after the excision and resumed his functional activity completely. Giant osteochondroma around the ankle is an extremely rare entity. Even rarer is a late presentation in the 6th decade onward. However, the management like others involves the excision of the lesion.


Percutaneous Ultrasound-Guided Superior and Inferior Mesenteric Vein Access for Portal Vein Recanalization-Transjugular Intrahepatic Portosystemic Shunt: A Case Series.
To describe the technique and outcomes of mesenteric access under ultrasound guidance to perform portal vein recanalization-transjugular intrahepatic portosystemic shunt (PVR-TIPS). Four patients (3 male: 1 female, mean age: 46.2 years; range 38-64 years) with portal vein thrombosis (PVT) and cavernous transformation were eligible for PVR-TIPS. Due to inaccessible splenic vein (one patient with history of splenectomy and 3 patients with unavailable splenic vein during the procedure), noninvasive direct puncture of superior (n = 3) and inferior (n = 1) mesenteric vein was conducted under ultrasound guidance to obtain access for PVR-TIPS. Trans-mesenteric access and PVR-TIPS were successful in all patients at first attempt. No immediate complication was observed following the procedures. Follow-up imaging with computed tomography (CT) scan and Doppler ultrasound revealed patent TIPS and portal venous vasculature in all patients. Percutaneous noninvasive transmesenteric access is a feasible approach for PVR-TIPS in patients with inaccessible splenic veins. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Increasing variance in North Pacific climate relates to unprecedented ecosystem variability off California.
Changes in variance are infrequently examined in climate change ecology. We tested the hypothesis that recent high variability in demographic attributes of salmon and seabirds off California is related to increasing variability in remote, large-scale forcing in the North Pacific operating through changes in local food webs. Linear, indirect numerical responses between krill (primarily Thysanoessa spinifera) and juvenile rockfish abundance (catch per unit effort (CPUE)) explained >80% of the recent variability in the demography of these pelagic predators. We found no relationships between krill and regional upwelling, though a strong connection to the North Pacific Gyre Oscillation (NPGO) index was established. Variance in NPGO and related central Pacific warming index increased after 1985, whereas variance in the canonical ENSO and Pacific Decadal Oscillation did not change. Anthropogenic global warming or natural climate variability may explain recent intensification of the NPGO and its increasing ecological significance. Assessing non-stationarity in atmospheric-environmental interactions and placing greater emphasis on documenting changes in variance of bio-physical systems will enable insight into complex climate-marine ecosystem dynamics.


Probing the donor and acceptor substrate specificity of the γ-glutamyl transpeptidase.
γ-Glutamyl transpeptidase (GGT) is a two-substrate enzyme that plays a central role in glutathione metabolism and is a potential target for drug design. GGT catalyzes the cleavage of γ-glutamyl donor substrates and the transfer of the γ-glutamyl moiety to an amine of an acceptor substrate or water. Although structures of bacterial GGT have revealed details of the protein-ligand interactions at the donor site, the acceptor substrate site is relatively undefined. The recent identification of a species-specific acceptor site inhibitor, OU749, suggests that these inhibitors may be less toxic than glutamine analogues. Here we investigated the donor and acceptor substrate preferences of Bacillus anthracis GGT (CapD) and applied computational approaches in combination with kinetics to probe the structural basis of the enzyme's substrate and inhibitor binding specificities and compare them with human GGT. Site-directed mutagenesis studies showed that the R432A and R520S variants exhibited 6- and 95-fold decreases in hydrolase activity, respectively, and that their activity was not stimulated by the addition of the l-Cys acceptor substrate, suggesting an additional role in acceptor binding and/or catalysis of transpeptidation. Rat GGT (and presumably HuGGT) has strict stereospecificity for L-amino acid acceptor substrates, while CapD can utilize both L- and D-acceptor substrates comparably. Modeling and kinetic analysis suggest that R520 and R432 allow two alternate acceptor substrate binding modes for L- and D-acceptors. R432 is conserved in Francisella tularensis, Yersinia pestis, Burkholderia mallei, Helicobacter pylori and Escherichia coli, but not in human GGT. Docking and MD simulations point toward key residues that contribute to inhibitor and acceptor substrate binding, providing a guide to designing novel and specific GGT inhibitors.


Use of CAD/CAM to fabricate duplicate abutments for retrofitting an existing implant prosthesis: a clinical report.
Computer-assisted design/computer-assisted manufacturing (CAD/CAM) technology has been widely used in the field of dentistry. Among CAD/CAM custom abutments, zirconia abutments are becoming more popular due to their favorable mechanical and esthetic properties. However, recent review articles show that fractured zirconia abutments are a common prosthetic complication. In this clinical report, a 72-year-old man presented with a dislodged implant-supported fixed prosthesis because of fractured zirconia abutments. This clinical report describes a successful application of CAD/CAM technology to fabricate duplicate abutments, which were retrofitted to the existing prosthesis.


Protocols to Assess Coagulation Following In Vitro Infection with Hemorrhagic Fever Viruses.
During the course of infection with a hemorrhagic fever virus (HFV), the checks and balances associated with normal coagulation are perturbed resulting in hemorrhage in severe cases and, in some patients, disseminated intravascular coagulopathy (DIC). While many HFVs have animal models that permit the analyses of systemic coagulopathy, animal infection models do not exist for all HFVs and moreover do not always recapitulate the pathology observed in human tissues. Furthermore, molecular analyses of how coagulation is affected are not always straightforward or practical when using ex-vivo animal-derived samples, thus reinforcing the importance of cell culture studies. This chapter highlights procedures utilizing human umbilical vein endothelial cells (HUVECs) as a model system to evaluate components of the intrinsic (prekallikrein (PK), factor XII (FXII), kininogen, and bradykinin (BK)) and extrinsic (Tissue Factor (TF)) systems. Specifically, protocols are included for the generation of a coculture blood vessel model, plating and infection of HUVEC monolayers and assays designed to measure activation of PK and FXII, cleavage of kininogen, and to measure the expression of TF mRNA and protein.


Radioimmunoassay of glycosphingolipids: application for the detection of forssman glycolipid in tissue extracts and cell membranes.
Development of a radioimmunoassay for detecting glycosphingolipids has been difficult, primarily because of transfer of radiolabeled glycolipid antigen to the unlabeled antigen pool. This difficulty has been overcome by the use of a radiolabeled glycolipid-polymer. Thus, an assay system has been developed for measuring picomolar quantities of Forssman hapten glycolipid. This assay is based on competition for rabbit anti-Forssman antibodies between Forssman glycolipid and a radiolabeled Forssman polyacrylic hydrazide polymer. Antigen-antibody complexes are removed quickly and efficiently by binding to formalin-fixed Staphylococcus aureus and subsequent centrifugation. One nanogram of Forssman glycolipid can be readily detected both in plasma membrane preparations and in purified glycolipid fractions. The isoantigenic expression of Forssman glycolipid in human gastrointestinal tissues has been reported previously (Hakomori et al., 1977). Using the radioimmunoassay, the Forssman status of several additional cases has been determined quantitatively.


The utility of SARS-CoV-2 genomic data for informative clustering under different epidemiological scenarios and sampling.
Clustering pathogen sequence data is a common practice in epidemiology to gain insights into the genetic diversity and evolutionary relationships among pathogens. We can find groups of cases with a shared transmission history and common origin, as well as identifying transmission hotspots. Motivated by the experience of clustering SARS-CoV-2 cases using whole genome sequence data during the COVID-19 pandemic to aid with public health investigation, we investigated how differences in epidemiology and sampling can influence the composition of clusters that are identified. We performed genomic clustering on simulated SARS-CoV-2 outbreaks produced with different transmission rates and levels of genomic diversity, along with varying the proportion of cases sampled. In single outbreaks with a low transmission rate, decreasing the sampling fraction resulted in multiple, separate clusters being identified where intermediate cases in transmission chains are missed. Outbreaks simulated with a high transmission rate were more robust to changes in the sampling fraction and largely resulted in a single cluster that included all sampled outbreak cases. When considering multiple outbreaks in a sampled jurisdiction seeded by different introductions, low genomic diversity between introduced cases caused outbreaks to be merged into large clusters. If the transmission and sampling fraction, and diversity between introductions was low, a combination of the spurious break-up of outbreaks and the linking of closely related cases in different outbreaks resulted in clusters that may appear informative, but these did not reflect the true underlying population structure. Conversely, genomic clusters matched the true population structure when there was relatively high diversity between introductions and a high transmission rate. Differences in epidemiology and sampling can impact our ability to identify genomic clusters that describe the underlying population structure. These findings can help to guide recommendations for the use of pathogen clustering in public health investigations.


Screening for cognitive impairment: a triage for outpatient care.
The current increase in aged individuals in number and proportion of the general population warrants dependable strategies to improve early detection of cognitive impairment. It was the goal of the present study to develop a triage for bedside testing and outpatient services. In a prospective clinical cohort study at the outpatient Memory Clinic, University of Ulm, Germany, 232 subjects were diagnosed with Alzheimer's disease [AD; NINCDS-ADRDA criteria; n = 66; age 65.9 +/- 7.3 years (mean +/- SD); Mini Mental State Examination (MMSE) score 23.4 +/- 4.1], mild cognitive impairment (MCI; criteria of Petersen et al.; n = 48; age 66.4 +/- 7.1 years; MMSE score 28.3 +/- 1.5), and major depressive disorder (DSM-IV criteria; n = 61; age 63.4 +/- 8.0 years; MMSE score 28.6 +/- 1.6). Diagnosis was secured with extensive neuropsychological, clinical, radiological, and laboratory investigations. Six brief screening tests including the Memory Impairment Screen (MIS), Letter Sorting Test (LST), Verbal Fluency (VF), and Clock Drawing Test (CDT) were assessed independently from the diagnostic procedure. We compared single items and composite scores. LST yielded a diagnostic accuracy of 0.81 and 0.62 for AD and MCI patients versus controls, respectively. With the MIS, diagnostic accuracy was 0.89 and 0.71, respectively. With a combination of LST, MIS, VF, and CDT, a sensitivity for AD and MCI patients of 1.00 and 0.83 was achieved. Thus, single-item screening (e.g. LST, VF) taking little more than 1 min and suitable for bedside testing or brief screening in the general practitioner's office yields diagnostic accuracy comparable to standard laboratory tests for other diseases. A composite of screening tests suitable for application in general outpatient care in neurological and psychiatric services reliably detects patients with AD and MCI.


Unilateral Versus Bilateral Percutaneous Pedicle Screw Fixation in Oblique Lumbar Interbody Fusion.
To compare clinical and radiologic outcomes of unilateral pedicle screw (UPS) and bilateral pedicle screw (BPS) fixations after single-level oblique lateral interbody fusion procedures. This retrospective study included 74 patients receiving single-level oblique lateral interbody fusion at L3-4 or L4-5 (June 2014 to June 2017). These patients were divided into BPS (n = 36) and UPS (n = 38) groups. Perioperative outcomes included operative time, blood loss, medical expenses, and complication rates. Radiologic outcomes included fusion rates and cage subsidence rates. Clinical outcomes included disability measured by the Oswestry Disability Index and pain measured by the visual analog scale. The operative time was significantly shorter in the UPS group (76.2 ± 4.4 minutes) compared with the BPS group (127.0 ± 5.6 minutes, P < 0.001). There was a significant between-group difference in medical expenses ($11,044.8 ± $470.7 in UPS group vs. $15,018.4 ± $547.1 in BPS, P < 0.001). Blood loss and hospital stay did not differ between groups. Oswestry Disability Index and visual analog scale scores significantly decreased in both groups at different time points postoperatively compared with preoperatively. The UPS group had lower Oswestry Disability Index and visual analog scale scores than the BPS group at 7 days postoperatively, but no between-group difference was found after 1-month follow-up. Fusion rate was 86.8% in UPS group and 91.7% in BPS group (P > 0.05). Cage subsidence rates did not differ between groups. Oblique lateral interbody fusion with UPS fixation is an effective and reliable option for single-level lumbar diseases. Compared with BPS fixation, it resulted in less blood loss, required less operative time, and had comparable effects on radiologic and clinical outcomes.


[Diagnostic procedure in chronic cervicofacial adenopathies].
Results of clinical examination are fundamental for establishment of an etiologic diagnosis, but the additional data provided by paraclinical investigations are essential elements. Adenectomy should be performed when etiology is uncertain.


Using INTERSPIA to Explore the Dynamics of Protein-Protein Interactions Among Multiple Species.
INTER-Species Protein Interaction Analysis (INTERSPIA) is a web application for identifying diverse patterns of protein-protein interactions (PPIs) in different species. Given a set of proteins of interest to the user, INTERSPIA first discovers additional proteins that are functionally associated with the input proteins as well as different or common patterns of PPIs among the proteins in multiple species through a server-side pipeline. Second, it visualizes the dynamics of PPIs in multiple species via an easy-to-use web interface. This article contains a basic protocol describing how to visualize diverse patterns of PPIs of input proteins in multiple species, and how to use them for functional analysis in the web interface. INTERSPIA is freely available at http://bioinfo.konkuk.ac.kr/INTERSPIA/. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Running INTERSPIA using a list of input proteins.


The basis of nuclear phospholipase C in cell proliferation.
Ca2+ is a highly versatile intracellular signal that regulates many biological processes such as cell death and proliferation. Broad Ca2+-signaling machinery is used to assemble signaling systems with a precise spatial and temporal resolution to achieve this versatility. Ca2+-signaling components can be organized in different regions of the cell and local increases in Ca2+ within the nucleus can regulate different cellular functions from the increases in cytosolic Ca2+. However, the mechanisms and pathways that promote localized increases in Ca2+ levels in the nucleus are still under investigation. This review presents evidence that the nucleus has its own Ca2+ stores and signaling machinery, which modulate processes such as cell proliferation and tumor growth. We focus on what is known about the functions of nuclear Phospholipase C (PLC) in the generation of nuclear Ca2+ transients that are involved in cell proliferation.


Expression, purification and crystallization of cysteine-rich human protein muskelin in Escherichia coli.
The increasing interest in the structural arrangements and functional interdependencies of individual modules within large multidomain proteins requires the development of new methods allowing efficient production and purification of large human proteins. Heterologous expression in bacteria is still the most convenient and widely-used approach. However, most of the existing tools are not well suited to expression of cysteine-rich proteins in a native-like soluble form, and with the increasing protein size refolding may result in obtaining non-native conformations or improper disulfide bridging pattern. Here, we present an efficient method of expression and purification of muskelin, a large, multidomain, cysteine-rich eukaryotic protein involved in cell adhesion and regulation of cytoskeleton dynamics. Using a broad range of purification and solubility tags, expression strains and conditions we optimized the procedure to acquire a natively folded protein of crystallization-scale quantity and purity. The correct protein conformation and disulfide bonding were anticipated from the results of circular dichroism spectra and Ellman's assay. Successful crystallization trials are a step towards muskelin crystal-structure determination, while the optimized expression and purification procedure can easily be applied to produce other eukaryotic proteins in the bacterial expression system.


Electrochemically driven extraction and recovery of ammonia from human urine.
Human urine contains high concentrations of nitrogen, contributing about 75% of the nitrogen in municipal wastewaters yet only 1% of the volume. Source separation of urine produces an ideal waste stream for nitrogen and phosphorus recovery, reducing downstream costs of nutrient treatment at wastewater treatment facilities. We examined the efficiency and feasibility of ammonia extraction and recovery from synthetic and undiluted human urine using an electrochemical cell (EC). EC processing of synthetic urine produced an ammonium flux of 384 ± 8 g N m(-2) d(-1) with a 61 ± 1% current efficiency at an energy input of 12 kWh kg(-1) N removed. EC processing of real urine displayed similar performance, with an average ammonium flux of 275 ± 5 g N m(-2) d(-1) sustained over 10 days with 55 ± 1% current efficiency for ammonia and at an energy input of 13 kWh kg(-1) N removed. With the incorporation of an ammonia stripping and absorption unit into the real urine system, 57 ± 0.5% of the total nitrogen was recovered as ammonium sulfate. A system configuration additionally incorporating stripping of the influent headspace increased total nitrogen recovery to 79% but led to reduced performance of the EC as the urine ammonium concentration decrease. Direct stripping of ammonia (NH3) from urine with no chemical addition achieved only 12% total nitrogen recovery at hydraulic retention times comparable with the EC systems. Our results demonstrate that ammonia can be extracted via electrochemical means at reasonable energy inputs of approximately 12 kWh kg(-1) N. Considering also that the hydrogen generated is worth 4.3 kWh kg(-1) N, the net electrical input for extraction becomes approximately 8 kWh kg(-1) N if the hydrogen can be used. Critical for further development will be the inclusion of a passive means for ammonia stripping to reduce additional energy inputs.


Taking Local Anesthesia to the Next Level: Four Strategies Clinicians May Consider.
The importance of effective local anesthesia delivery in dentistry cannot be overstated, as many dental procedures could not be performed without it. Despite the availability to oral healthcare professionals of various successful local anesthetics, the administration of local anesthesia continues to provoke anxiety among dental patients and remains an often unpleasant experience. A number of techniques may be utilized to help mitigate injection pain and improve the patient experience. This article reviews four strategies clinicians may consider to increase the effectiveness of their delivery of local anesthesia: warming or cooling of local anesthetic solutions, vibration and distraction techniques, liposomal encapsulation, an buffering or alkalinization of dental local anesthetics.


First-line Chemotherapy Responsiveness and Patterns of Metastatic Spread Identify Clinical Syndromes Present Within Advanced KRAS Mutant Non-Small-cell Lung Cancer With Different Prognostic Significance.
Unsuccessful KRAS-specific treatment approaches in non-small-cell lung cancer (NSCLC) might reflect underlying disease heterogeneity. We sought to define clinical "syndromes" within advanced KRAS mutant NSCLC to improve future clinical trials and create a clinical framework for future molecular development. To test a series of a priori hypotheses regarding KRAS-mutant NSCLC clinical syndromes, we conducted a multi-institutional retrospective medical record review. Survival probabilities were estimated using the Kaplan-Meier model. Between-group differences were assessed using the log-rank test. Multivariate Cox regression analyses and Wilcoxon rank sum testing were used to assess progression-free survival and overall survival (OS) differences. Among 218 patients with advanced KRAS-mutant NSCLC, OS and progression-free survival with first-line chemotherapy did not differ by intrathoracic versus extrathoracic spread, smoking intensity, or the specific KRAS mutation. Metastatic disease at diagnosis resulted in significantly worse OS than recurrent, unresectable disease (median OS, 14.6 vs. 40.9 months; P = .001). Among the patients with metastatic disease at diagnosis, nonscalp, soft tissue metastases (syndrome X; 6% of cases; 95% confidence interval [CI], 2.5%-10.1%) signified a poor prognosis (median OS, 7.5 vs. 15.9 months for the controls; P = .021). The response to first-line chemotherapy (syndrome Y; 41% of cases; 95% CI, 32.3%-50.6%) signified a good prognosis (median OS, 26.7 vs. 11.9 months; P = .002). The overlap between these 2 syndromes was minimal (2 of 111). Multivariate analysis confirmed these observations. The hazard ratio for death for syndromes X and Y was 2.64 (95% CI, 1.13-6.14) and 0.45 (95% CI, 0.28-0.76), respectively. Chemotherapy-responsive disease and nonscalp, soft tissue spread might represent distinct clinical syndromes within KRAS-mutant NSCLC. The molecular biology underlying this heterogeneity warrants future studies.


The antiangiogenic agent neovastat (AE-941) inhibits vascular endothelial growth factor-mediated biological effects.
Vascular endothelial growth factor (VEGF) is a potent regulator of angiogenesis, which exerts direct effects on vascular endothelial cells, including endothelial cell proliferation and survival, tubulogenesis, and vascular permeability. In this study, we examined whether Neovastat, a naturally occurring multifunctional antiangiogenic drug, could inhibit the endothelial cell response to VEGF stimulation. We demonstrated that Neovastat was able to block the VEGF-dependent microvessel sprouting from Matrigel-embedded rat aortic rings, and it also blocked the VEGF-induced endothelial cell tubulogenesis in vitro. In vivo studies showed that Neovastat was able to specifically inhibit VEGF-induced plasma extravasation in numerous tissues, including pancreas and skin. The mechanism of action of Neovastat on VEGF-mediated effects was also evaluated at the molecular level. Neovastat was shown to compete against the binding of VEGF to its receptor in endothelial cells and significantly inhibited the VEGF-dependent tyrosine phosphorylation of VEGF receptor-2, whereas it had no significant effect on VEGF receptor-1 activity. Moreover, the inhibition of receptor phosphorylation was correlated with a marked decrease in the ability of VEGF to induce pERK activation. Neovastat does not compete against the binding of basic fibroblast growth factor, indicating a preferential inhibitory effect on the VEGF receptor. Because Neovastat was shown previously to inhibit metalloproteinase activities, these results suggest that Neovastat is able to target multiple steps in tumor neovascularization, further emphasizing its use as a pleiotropic, multifunctional antiangiogenic drug.


Translating instruments into other languages: development and testing processes.
Cross-cultural influences affect perceptions and health practices, which are 2 areas of nursing concern. Culturally sensitive assessment instruments are needed, but many challenges exist in obtaining valid and reliable measurement. Translating questionnaires for cross-cultural research is fraught with methodological pitfalls related to colloquial phrases, jargon, idiomatic expressions, word clarity, and word meanings. It cannot be assumed that a particular concept has the same relevance across cultures. Simply translating an English version word-for-word into another language is not adequate to account for linguistic and cultural differences. Ideally, the perspectives of people from the culture about the concept of interest should be studied first, but often a practical alternative is to find and translate a tool developed in another culture. The purpose of this article is to describe important considerations in conducting translation for equivalence, types of equivalence, and strategies to translate instruments that promote equivalence and how to test the translated version for equivalence. These concepts and strategies are illustrated by describing the translation process of Hilton's Uncertainty Stress Scale into French and the use and testing of the French version with a French Canadian sample in Skrutkowski's study of perceived uncertainty in adult survivors of cancer.


(2,7-Di-meth-oxy-naphthalen-1-yl)(naph-thalen-1-yl)methanone.
The asymmetric unit of the title compound, C23H18O3, contains two independent mol-ecules (A and B). Each mol-ecule has essentially the same conformation (r.m.s. deviation of fitted mol-ecules = 0.173 Å) with the aromatic rings twisted in a near perpendicular manner. The dihedral angles between the two naphthalene ring systems are 79.07 (4) and 88.19 (4)° in the two independent mol-ecules. In the crystal, the A mol-ecules are connected by C-H⋯O inter-actions, forming chains along the b-axis direction. Further C-H⋯O inter-actions between the H atoms of the meth-oxy group and the O atoms of the carbonyl units link the A and B mol-ecules, forming a three-dimensional network.