Leaf and environmental parameters influencing transpiration: Theory and field measurements.
The influence of variations in the boundary air layer thickness on transpirtion due to changes in leaf dimension or wind speed was evaluated at a given stomatal resistance (r s) for various combinations of air temperature (T a) and total absorbed solar energy expressed as a fraction of full sunlight (S ffs). Predicted transpiration was found to either increase or decrease for increases in leaf size depending on specific combinations of T a, S ffs, and r s. Major reductions in simulated transpiration with increasing leaf size occurred for shaded, highly reflective, or specially oriented leaves (S ffs=0.1) at relatively high T a when r s was below a critical value of near 500 s m-1. Increases in S ffs and decreases in T a lowered this critical resistance to below 50 s m-1 for S ffs=0.7 and T a=20°C. In contrast, when r s was above this critical value, an increase in leaf dimension (or less wind) resulted in increases in transpiration, especially at high T a and S ffs. For several combinations of T a, S ffs, and r s, transpiration was minimal for a specific leaf size. These theoretical results were compared to field measurements on common desert, alpine, and subalpine plants to evaluate the possible interactions of leaf and environmental parameters that may serve to reduce transpiration in xeric habitats.


Nutritional status of lacto-ovo vegetarian Trappist monks.
The nutritional status of members of a lacto-ovo vegetarian Trappist community was studied. Body weights and heights were normal. An analysis of four weeks' menus showed that, for the most part, sufficient nutrients were provided. A 24-hr. dietary recall revealed that a number of the subjects had low intakes of some nutrients, particularly the B-vitamins and calcium, iron, magnesium, and zinc. Improvement in the nutritional intakes and resulting nutritional status of the men must rely on an availability of accurate educational materials and each individual's determination to gain the knowledge needed to make a wiser selection of food.


Characterization of cat messenger RNA decay suggests that turnover occurs by endonucleolytic cleavage in a 3' to 5' direction.
Turnover of the chloramphenicol acetyltransferase (cat) messenger RNA in Escherichia coli was investigated by analysis of cellular mRNA decay intermediates and the transcript sequence. Analysis of the sequence has revealed the presence of a repetitive extragenic palindromic (REP) element at the 3' end of the transcript as well as several 5'-UUAU-3' sequences, two elements which have roles in modulating turnover of other E. coli mRNAs. For cat mRNA, however, removal of the REP sequence has no effect on the half-life of the transcript, indicating that the REP sequence does not stabilize the upstream region of this message. Results from mapping of the mRNA decay products by several techniques suggest that the message is instead subject to endonucleolytic attack at five sites 5' of the REP element. The sequence UUAU is present at three of these five sites. It also appears that the cat mRNA is sequentially cleaved in a 3' to 5' direction during turnover of this mRNA in vivo. A model for cat mRNA turnover is discussed.


A Comparative Study of Associated Microbiota Between Pig Farm and Pig Slaughterhouse in Guangdong, China.
The goal of this study was to compare the microbiota in different pig-present settings in China. Bioaerosol samples from pig farms and slaughterhouses and nasal samples from pig farmers and slaughterhouse workers were collected in Guangdong, southern China. The bacterial genomic DNA was isolated and subjected to 16S sequencing. The data were analyzed using QIIME2 with the DADA2 pipeline. A total of 14,923,551 clean reads and 2785 operational taxonomic units (OTUs) were obtained, which were mostly grouped into 4 phyla (Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria) and 220 families. The microbiota richness of nasal samples in pig-present workers was higher than that of bioaerosols collected in the vicinity of the pig enclosures. There were 31.7% (620/1954) shared OTUs between pig farm bioaerosols and pig farmers which was higher than that between pig slaughterhouses and slaughterhouse workers (23.4%, 364/1553) (p < 0.001). Acinetobacter and Pseudomonas were the most abundant in pig-present bioaerosols, and Staphylococcus, Pseudomonas, and Corynebacterium were dominant bacterial genus in pig farmers. The bacterial patterns are also specific to the location of sample collected. The results suggest that bioaerosol microbiota interact with human nasal microbes in the vicinity of the pig farm enclosures, providing the basis for further analysis of microbial transmission across hosts in pig-present settings.


Effect of neoadjuvant chemotherapy on locally advanced upper tract urothelial carcinoma: a pooled analysis.
The outcome of neoadjuvant chemotherapy (NAC) has been established in bladder cancer but remains controversial in upper tract urothelial carcinoma (UTUC). In this work, we explored the therapeutic effect of NAC in patients with locally advanced UTUC. We conducted a literature search on articles published from 1995 up to April 2020 in PubMed/Medline, the Cochrane Library, Embase, Google Scholar. A total of 19 eligible studies with 6,283 patients were identified, from which the overall survival (OS), cancer-specific survival (CSS), progression-free survival (PFS), disease-free survival (DFS), pathological complete response (pCR) rate and pathological partial response (pPR) rate were extracted. All analyses were conducted using Review Manager 5.3 and Stata statistical software (version 15). In total, 6,283 UTUC patients were included from 19 eligible studies out of which 1,474 patients received NAC and subsequent radical nephroureterectomy (RNU), whereas 4,809 patients received RNU only. Compared with single RNU, patients with NAC and subsequent RNU exhibited longer OS, CSS, PFS, DFS by hazard ratio (HR) 2.14 [95% confidence interval (CI): 1.75-2.63; P<0.001], HR 2.07 (95% CI: 1.49-2.87; P<0.001), HR 2.00 (95% CI: 1.42-2.83; P<0.001), and HR 3.76 (95% CI: 2.16-6.56; P<0.001). pCR rate and pPR rate of NAC are 0.10 (0.07-0.13) and 0.40 (95% CI: 0.32-0.49, P <0.001) respectively. This work revealed that NAC and subsequent RNU provided better survival outcomes in patients with locally advanced UTUC when compared with single RNU.


Sensitivity analysis for random measurement error using regression calibration and simulation-extrapolation.
Sensitivity analysis for random measurement error can be applied in the absence of validation data by means of regression calibration and simulation-extrapolation. These have not been compared for this purpose. A simulation study was conducted comparing the performance of regression calibration and simulation-extrapolation for linear and logistic regression. The performance of the two methods was evaluated in terms of bias, mean squared error (MSE) and confidence interval coverage, for various values of reliability of the error-prone measurement (0.05-0.91), sample size (125-4000), number of replicates (2-10), and R-squared (0.03-0.75). It was assumed that no validation data were available about the error-free measures, while correct information about the measurement error variance was available. Regression calibration was unbiased while simulation-extrapolation was biased: median bias was 0.8% (interquartile range (IQR): -0.6;1.7%), and -19.0% (IQR: -46.4;-12.4%), respectively. A small gain in efficiency was observed for simulation-extrapolation (median MSE: 0.005, IQR: 0.004;0.006) versus regression calibration (median MSE: 0.006, IQR: 0.005;0.009). Confidence interval coverage was at the nominal level of 95% for regression calibration, and smaller than 95% for simulation-extrapolation (median coverage: 85%, IQR: 73;93%). The application of regression calibration and simulation-extrapolation for a sensitivity analysis was illustrated using an example of blood pressure and kidney function. Our results support the use of regression calibration over simulation-extrapolation for sensitivity analysis for random measurement error.


Chronic Fatigue Syndrome: An Evaluation of a Community Based Management Programme for Adolescents and their Families.
Young people with chronic fatigue syndrome (CFS), families and clinicians may differ in their attributions about CFS and consequently in their approach to treatment. Research that clarifies the best treatment approaches is clearly needed. We have sought to develop a model that engages young people and their families in a collaborative way. The approach adopts an optimistic and holistic stance using an active rehabilitation model paying attention to the integrated nature of the physiological and psychological aspects of the illness. This small study set out to evaluate this approach from a service user perspective. Semi-structured interviews were carried out with young people and their parents separately in order to elicit their views on key treatment elements and their perceived degree of recovery. Improvements are indicated in all key areas addressed and qualitative information suggests that families value this approach. Further research is needed to address treatment issues for families who choose not to opt into the service model.


Formaldehyde adducts of glutathione. Structure elucidation by two-dimensional n.m.r. spectroscopy and fast-atom-bombardment tandem mass spectrometry.
Aqueous mixtures of formaldehyde and glutathione react to form a variety of cyclized adducts in addition to S-hydroxymethylglutathione. The adducts are in labile equilibrium with each other and are not readily separated. The structures of two of the other major adducts were determined by concerted application of 13C-1H two-dimensional chemical-shift correlation, fast-atom-bombardment mass spectrometry and tandem mass spectrometry to the adduct mixtures in aqueous solution.


Effects of water restriction and dirt on grooming behavior in neotropical rodents (Trinomys setosus and T. yonenagae) (Echimyidae).
Grooming in rodents presents an evolutionarily conserved behavioral pattern that may cause water loss since saliva is deposited during washing onto large body surfaces. Trinomys yonenagae and Trinomys setosus are sister species of spiny rats occurring in Brazil, the former inhabiting a paleodesert of fixed dunes in the Caatinga, the latter being found in mesic environments of the Atlantic Forest. Consequently, it is expected that both species evolved under different selective pressures related to water balance, with T. yonenagae presenting mechanisms for dealing with water deprivation not found in T. setosus. Reduction of self-cleaning expression seems to offer a possible way to save water, as previously suggested by studies of the sand-dwelling spiny rat. Therefore, we propose to investigate grooming under four conditions: 'control' (C), a regimen of 'water restriction' (WR), of 'dirt' (D), and the combination of both conflicting stimuli (WR + D), in T. setosus, T. yonenagae, and Rattus norvegicus to compare the behavioral responses of these species. The main differences are observed in the forest dweller: T. setosus expresses a low relative duration of face washing under C, whose value is intermediate between the ones found in the two other species. WR treatment does not alter this pattern, however, the addition of dirt (D, WR + D) significantly increases the relative duration of washing in relation to C. Locomotor activity is decreased both in T. setosus and Wistar rats when they are under WR, a situation that could jeopardize antipredatory performance. T. yonenagae, the sand dweller, maintains a significantly lower expression of washing under C, as previously suggested, and under WR, D and WR + D. In addition, differently from the other two species the sand dweller maintains a normal activity level during all treatments. This study suggests differences in grooming as a strategy alluding to water balance by the two spiny rats inhabiting different ecosystems. A significantly clear pattern that saves water is observed in T. yonenagae, which probably has contributed to his evolution in one of the hottest semiarid areas of the world.


Involvement of mitogen-activated protein kinase (MAPK) pathway in LH- and meiosis-activating sterol (MAS)-induced maturation in rat and mouse oocytes.
Gonadotropic stimulation of meiotic resumption in mice is dependent upon mitogen-activated protein kinase (MAPK) activation in the somatic compartment of the follicle. By contrast, spontaneous resumption of meiosis is independent of MAPK activation. In view of the suggested role of meiosis-activating sterol (MAS) in oocyte maturation we have (i) compared MAPK activation in rat preovulatory follicles stimulated by LH or by accumulation of endogenous MAS by using an inhibitor of MAS conversion, AY9944; (ii) examined whether stimulation of meiosis by MAS is dependent upon MAPK activation using denuded oocytes (DO) of Mos- null mice (hereafter Mos(-/-)) with oocytes unable to activate MAPK. Rat preovulatory follicles responded to LH or AY9944 stimulation by MAPK activation. Inhibition of MAPK phosphorylation blocked both LH- and AY9944 triggered resumption of meiosis. In mouse cumulus-enclosed oocytes (CEOs) and DOs AY9944 stimulated GVB in wild-type and Mos(-/-) mouse CEOs cultured with hypoxanthine (Hx). Addition of MAS or AY9944 to mouse DOs cultured with Hx induced resumption of meiosis only in wild-type and Mos(+/-) oocytes, but they were ineffective in Mos(-/-) oocytes. The observed sluggish activation of MAPK induced by AY9944 in rat follicle-enclosed oocytes (FEO) may cause the delay in meiotic resumption in response to MAS and AY9944 stimulation. Further, it is incompatible with the suggested role of MAS as an obligatory mediator of LH in the induction of meiotic maturation. MAPK/MOS activation, whether in the somatic compartment or in denuded oocytes, is required for MAS- like LH-, FSH-, or EGF-induced resumption of meiosis.


Effect of acute administration of nicotine on in vivo release of noradrenaline in the hippocampus of freely moving rats: a dose-response and antagonist study.
The effect of systemic administration of (-)-nicotine on release of noradrenaline in the hippocampus was studied by in vivo microdialysis in freely moving rats, using dialysate containing nomifensine (5 microM). (-)-Nicotine, at both 0.4 and 0.8 mg/kg but not 0.2 mg/kg, rapidly and significantly increased extracellular levels of noradrenaline. Extracellular levels of dopamine were also increased, but this was only significant after the larger dose. Both 0.4 and 0.8 mg/kg also produced a significant increase in extracellular levels of the metabolites of dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid. Extracellular levels of the metabolite of 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, increased after 0.8 mg/kg but this effect was only apparent much later. Injection of a second 0.8 mg/kg challenge of (-)-nicotine, 150 min after the first, produced similar increases in extracellular levels of noradrenaline, dopamine, 3-4-dihydroxyphenylacetic acid and homovanillic acid. Over the experimental period, there was no further increase in extracellular levels of 5-hydroxyindoleacetic acid. Increases in extracellular levels of noradrenaline, dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid, in response to 0.8 mg/kg (-)-nicotine, were prevented by the systemic administration of mecamylamine, but not hexamethonium (both at 5 mg/kg). Mecamylamine also inhibited the delayed increase in extracellular levels of 5-hydroxyindoleacetic acid, produced by the first injection of (-)-nicotine. These results suggest that (-)-nicotine, dose-dependently stimulated the release and metabolism of amine transmitters by an action at central nicotinic receptors. However, the precise site of action, i.e. at nerve terminals, cell bodies or both, requires further elucidation.


Endoplasmic Reticulum Stress of Oral Squamous Cell Carcinoma Induces Immunosuppression of Neutrophils.
The endoplasmic reticulum (ER) stress of cancer cells not only determined cancer cell fate but also indirectly triggered proinflammatory or immunosuppressive responses of macrophages. In addition, ER stressed neutrophils were known to acquire immunosuppressive activity with surface expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1). Since the importance of tumor ER stress and immunosuppressive neutrophils has been emphasized in head and neck cancers, we hypothesized that the ER stress of oral squamous cell carcinoma (OSCC) could transform neutrophils into LOX-1 expressing immunosuppressive phenotype. Two human OSCC cell lines, SCC25 and OML1, were treated with either vehicle or thapsigargin (THG), an ER stress inducer. These tumor conditioned media (TCM) were collected accordingly. Then human peripheral blood neutrophils from healthy donors were cultured in these TCM. The results showed that neutrophils cultured in THG-treated TCM had higher expression of LOX-1 compared with those cultured in vehicle-treated TCM. Moreover, by interleukin-2/anti-CD3/anti-CD28 activated autologous T cell proliferation assay, neutrophils conditioned by THG-treated TCM were shown to inhibit T cell proliferation more significantly than those conditioned by vehicle-treated TCM. These novel findings indicated that the ER stress of OSCC could be transmitted to neutrophils which in turn expressed LOX-1 and obtained immunosuppressive ability. Our findings further supported the existence of "transmissible" ER stress between tumor cells and neutrophils.


Microcystic adenoma of the pancreas (glycogen-rich cystadenoma) with stromal amyloid deposits.
We report a case of a pancreatic glycogen-rich microcystic serous adenoma with stromal amyloid deposits, focusing on the significance of isolated amyloid deposits in tumours. The architectural pattern was characterized by thin-walled cysts lined by a single layer of flat or cuboidal epithelial cells intensely stained by the PAS-reaction only before diastase digestion, suggesting the presence of glycogen. Tumour stroma was composed of broad fibrocollagenous tissue with lamellar hyalinized areas which were positively stained by Congo red and showed green birefringence and dichroism with polarized light. For amyloid protein characterization, immunohistochemical studies were performed with anti-beta amyloid protein and anti-amyloid precursor pre-A4695. The former antibody diffusely stained tumour stroma, while the latter stained only scattered stroma cells. This is the first documented case of amyloid deposition in pancreatic serous adenoma. We indicate that the source of amyloid is an APP-like precursor secreted by stromal myofibroblasts.


Association between serum uric acid and carotid disease in patients with atherosclerotic acute ischemic stroke.
The role of serum uric acid in ischemic stroke is controversial. On the one hand, it has a role as neuroprotectant in acute phase, but on the other hand, it may promote atherosclerosis in carotid arteries. Our aim is to investigate the association of serum uric acid levels at admission of acute ischemic stroke patients with carotid disease. Cross-sectional study of patients admitted due to acute ischemic stroke. Clinical and laboratory variables were recorded. The carotid disease was defined based on the findings of carotid echography intima to media thickness and stenosis. Patients were grouped according to these findings. Robust statistical methods were applied into analysis. A total of 245 patients were recruited through a stroke registry. Their values of serum uric acid were related to both carotid intima to media thickness and stenosis showing a positive relationship between serum uric acid levels and intima to media thickness by Pearson correlation ( p < 0.05). Similarly, after adjusting for all potential confounders, eGFR, glucose, age and serum uric acid levels, (OR 1.26 (95% CI 1.04 -1.52, p¼0.01), were identified as independent predictors for having a intima to media thickness ≥1 mm. Similarly, by grouping patients in tertiles of the serum uric acid distribution, we found a predominantly greater carotid disease in the tertile with the highest levels of serum uric acid ( p < 0.005). Our study supports the hypotheses that serum uric acid levels have different roles in the case of ischemic stroke. Its effects on the vascular wall contribute to the development of atherosclerosis and carotid disease.


Metabolic inflexibility and unusual catabolism in Leigh-like syndrome due to m.10191T>C.
Hypercatabolism is a well-known feature of mitochondrial diseases but some patients may present with hypometabolism, as the following case. Case report using standard investigation methods. The patient is a 32 years-old female with a Leigh-like syndrome due to the mtDNA variant m.10191 T > C in MT-ND3. Leigh-like syndrome is characterized by symmetric basal ganglia or brainstem lesions plus involvement of organs other than the brain. The patient presented with hypometabolism, which did not respond to ketogenic diet but responded to fasting. The patient showed a Warburg-like effect, which resulted in reliance on glucose due to the exclusion of oxidative phosphorylation with an extremely low VO2max. The patient only entered substantial ketosis when all gluconeogenic substrates were removed. Prolonged survival in the index patient may have possibly resulted from this previously unreported protective mechanism to reduce oxidative stress. The unusual Warburg-like phenomenon was interpreted as a possible mechanism of patients with a mitochondrial disease to survive into adulthood. This case shows that mitochondrial disease can manifest with hypometabolism and that an unusual Warburg-like effect may be responsible in some patients with mitochondrial disease to survive into adulthood.


Thymosin β4 attenuates liver fibrosis via suppressing Notch signaling.
Thymosin β4 (Tβ4) has been shown to have beneficial effects in a number of pathological processes. Although there was research reporting the endogenous expression of Tβ4 influences hepatic stellate cells (HSCs) activation, the effect of exogenous administration of Tβ4 in hepatic fibrosis remains unclear. In the current study, we used CCl4-induced liver fibrosis model mice to investigate the effect of Tβ4 administration on fibrosis in vivo and the underlying mechanism. Our study indicates that Tβ4 attenuates hepatic fibrosis and down-regulates the expression of fibrogenic genes in hepatic liver. In addition, Tβ4 inhibits the expression of pro fibrogenic and proliferation genes in activated HSCs. Further study revealed that Tβ4 attenuates liver fibrosis through inhibition of the Notch signaling, as Tβ4 significantly reduces the expression of Notch2 and Notch 3 that were increased in hepatic liver. Our data indicate that Tβ4 might be an effective anti-fibrotic drug for the treatment of liver fibrosis.


Neurodevelopmental Disorders: Sensing Tourette's Tics Away.
Though still shrouded in mystery, Gilles de la Tourette's syndrome is widely regarded as an archetypal neurodevelopmental disorder of central, motor control. New evidence that its cardinal manifestation - prominent tics - may be ameliorated by a peripheral, sensory intervention compels us to revise not only our conception of the syndrome, but of the motor system itself.


D-Mannose ameliorates DNCB-induced atopic dermatitis in mice and TNF-α-induced inflammation in human keratinocytes via mTOR/NF-κB pathway.
D-mannose is a C-2 epimer of glucose, widely distributed in nature. Atopic dermatitis (AD) is a chronic inflammatory disease characterized by repetitious itching. The present study aimed to explore the protective effect and the underlying mechanism of D-mannose against the development of atopic dermatitis. We tested the effect of D-mannose by establishing DNCB (2,4-dinitrochlorobenzene)-induced AD mice models in vivo and culturing keratinocytes (HaCaT and NHEK) in vitro. The skin lesion severity was evaluated by histochemical staining. Cytokine expression levels were measured by real-time PCR and ELISA assay. The expression of the mammalian target of rapamycin (mTOR)/ nuclear transcription factor κB (NF-κB)-signaling-related molecules were determined by western blotting. Here, we found that topical supplementation of D-mannose remarkably attenuated skin lesions and recovered skin barrier function in AD mice model induced by DNCB. Furthermore, in vivo and in vitro experiments indicated that D-mannose inhibited tumor necrosis factor-α (TNF-α)-mediated increased expression of inflammatory cytokines. D-mannose also markedly downregulated TNF-α-stimulated activation of mTOR/NF-κB signaling pathway that was crucial for regulating the inflammatory condition. However, these effects were abolished by treatment with inhibitors of mTOR or NF-κB in HaCaT and NHEK. As far as we know, this is the first study uncovering the effective role of D-mannose via skin topical application. We found that D-mannose plays a regulatory role on inflammatory keratinocytes, suggesting its therapeutic utilization as a potential drug against atopic dermatitis.


Cloning, in vitro expression and bioactivity of duck interleukin-18.
The encoding sequence for duck IL-18 was obtained, using reverse transcription-polymerase chain reaction, from mRNA harvested from Con A-stimulated Gushi (GS) duck splenic mononuclear cells. Recombinant duck IL-18 (rduIL-18) was produced in a prokaryotic expression system. In vitro bioactivity of rduIL-18 was determined in a lymphocyte proliferation assay and in vivo bioactivity of rduIL-18 was assessed by addition to a vaccine. Monoclonal antibody (mAb) and polyclonal antibodies (pAbs) specific for rduIL-18 were generated and subsequently characterized by ELISA, Western blot and neutralizing assays. Sequence analysis of GS duck IL-18 demonstrated an open reading frame (ORF) of 603 base pairs encoding for a 200 amino acid precursor protein. The duck encoding sequence shares 85.3% similarity to the chicken equivalent, at the nucleotide level. A His-duIL-18 fusion protein was recognized in Western blot by mAbs against duck and chicken IL-18 (chIL-18), but not by mAb against human IL-18. Recombinant duIL-18 induced in vitro proliferation of Con A-stimulated duck splenocytes and enhanced the immune response of ducks vaccinated with an inactivated oil emulsion vaccine against avian influenza virus. PAb and mAb 5B2 against rduIL-18 had neutralizing ability, inhibiting the biological activities of both recombinant duIL-18 and endogenous duIL-18. The results indicate that rduIL-18 has the potential to be used as an immunoadjuvant, and the mAb against rduIL-18 further facilitates basic immunobiological studies of the role of IL-18 in the avian immune system.


Technique for placement of peritoneovenous shunts in presence of abdominal adhesions.
Abdominal adhesions may cause kinking of the peritoneal limb of peritoneovenous shunts during placement. Use of a stylet in the peritoneal limb can facilitate placement and avoid kinking that could lead to shunt dysfunction.


Prenatal diagnosis of Beckwith-Wiedemann syndrome.
The diagnosis of Beckwith-Wiedemann syndrome (BWS) typically is made after birth. To our knowledge, no established guidelines exist for the prenatal diagnosis of this condition. We present two new cases of prenatally diagnosed BWS and propose a diagnostic schema. Published reports obtained from Medline searches were reviewed and combined with our cases. Our proposed schema of two major criteria (abdominal wall defect, macroglossia, macrosomia) or one major plus two minor criteria (nephromegaly/dysgenesis, adrenal cytomegaly, aneuploidy/abnormal loci, polyhydramnios) fits all published reports of prenatally diagnosed BWS. The prenatal diagnosis of BWS can be reliably made by applying our proposed guidelines. This schema allows for uniform fetal diagnosis of the syndrome and helps prepare for prenatal counseling and peri- and post-natal management strategies.


Calcium imaging of multiple neurons in freely behaving C. elegans.
Caenorhabditis elegans is a popular model organism to study how neural circuits and genes regulate behavior. To reliably correlate circuit function with behavior, it is important to record neuronal activity in freely behaving worms. As neural circuits are composed of multiple neurons that cooperate to process information, it is highly desirable to simultaneously record the activity of multiple neurons in the circuitry. However, such a system has not been available in C. elegans. Here, we report the CARIBN II (Calcium Ratiometric Imaging of Behaving Nematodes version II) system. This system provides smoother data collection and more importantly permits simultaneous imaging of calcium transients from multiple neurons in freely behaving worms. Using this system, we imaged the activity of AVA and RIM, two key neurons in the locomotion circuitry that regulate backward movement (reversal) in locomotion behavior. We found that AVA activity increases while RIM activity decreases during the same reversal events in spontaneous locomotion, consistent with the recent report that the AVA and RIM are involved in promoting the initiation of reversals. The CARIBN II system provides a valuable tool for dissecting the neural basis of behavior in C. elegans.


Prognosis is similar for patients who undergo primary PCI during regular-hours and off-hours: A report from SCAAR.
Timely percutaneous coronary intervention (PCI) improves prognosis in ST-elevation myocardial infarction (STEMI). However, recent reports indicate that patients with STEMI who present during non-regular working hours (off-hours) have a worse prognosis. The aim of this study was to compare outcome between patients with STEMI who underwent primary PCI during off-hours and regular hours. We retrieved data from the Swedish Coronary Angiography and Angioplasty Registry (SCAAR) for all patients who underwent primary PCI in Region Västra Götaland due to STEMI between January 2004 and May 2013. We modeled unadjusted and adjusted Cox proportional-hazards regression and logistic regression models for the outcomes death, cardiogenic shock, stent thrombosis and in-stent restenosis. A propensity score-adjusted Cox proportional-hazards model, which adjusted for traditional cardiovascular risk factors was predefined as the primary statistical model. Death at any time during the study period was pre-specified as primary end-point. During the study period 4.611 (65%) patients underwent primary PCI due to STEMI during off-hours and 2,525 (35%) during regular hours. The risk of dying was similar among the groups for the primary endpoint death at any time during the study period (HR 1.00, 95% CI 0.89-1.12, P = 0.991) and for secondary end-point death within 30 days (HR 1.03; 95% CI 0.85-1.25, P = 0.735). The risks of developing cardiogenic shock, stent thrombosis, or in-stent restenosis were similar between the groups. In our region, short- and long-term prognosis for patients with STEMI who undergo primary PCI is similar for patients presenting during off-hours and regular hours.


General practice management of overweight and obesity in children and adolescents in Australia.
Childhood obesity is rapidly increasing in prevalence worldwide, but healthcare capacity to address this problem seems limited. The purpose of this study was to describe the prevalence and rate of management of childhood overweight and obesity in Australian general practice. A cross-sectional study consisting of 3978 general practitioners (GPs), randomly selected using Medicare Australia claims, who recorded 42,515 encounters with children age 2-17 including 12,925 sub-sampled encounters with self or carer-reported height and weight collected. Prevalence of overweight and obesity, rate of management of overweight and obesity, content of encounters in overweight and nonoverweight children, content of encounters in those managed for overweight and obesity, and management to prevalence ratio. A total of 29.6% of sub-sampled children were classified as overweight (18.3%) or obese (11.4%). GPs managed overweight and obesity during 215 encounters, or once per 200 encounters with children age 2-17 and once per 58 encounters with overweight or obese children. The content of encounters in overweight and non-overweight children did not differ. Children who were managed for overweight or obesity presented with these conditions as reasons for the encounter significantly more often [66.5 (95% confidence interval (CI): 59.7-73.3) vs. 1.2 (95% CI: 1.0-1.3)] and were managed for more problems, particularly depression [4.2 (95% CI: 1.5-6.9) vs. 0.8 (95% CI: 0.7-0.9)], than average per 100 encounters. Consultations for overweight or obesity were significantly longer than average [16.7 (95% CI: 14.7-18.7) vs. 12.4 (95% CI: 12.2-12.5) minutes]. Overweight and obesity are prevalent in children presenting to Australian general practice but GPs do not use most of the available opportunities to manage this problem.


Gluconeogenic key enzymes in normal and intrauterine growth-retarded newborn rats.
Intrauterine growth retardation was induced in rats by ligation of the artery of one of both uterine horns. Activities of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, fructose-1,6-diphosphatase and glucose-6-phosphatase in liver were measured at 0, 1, 3 and 6 h after delivery in newborn rats from normal and sham-operated litters, and from ligated and contralateral uterine horns. Lower activities of fructose-1,6-diphosphatase were found in small-for-gestational-age animals in comparison with animals from contralateral horns. When small-for-gestational-age animals were compared with animals from sham litters (which could be regarded as more satisfactory controls), the activities of two other gluconeogenic enzymes (pyruvate carboxylase and glucose-6-phosphatase) appeared to be lower as well. It is concluded that a delay in the development of these gluconeogenic enzymes could play a role in neonatal hypoglycemia in small-for-gestational-age rats.


Sporadic late onset nemaline myopathy with concurrent dermatological symptoms responding to immunosuppressive treatment.
Sporadic late onset nemaline myopathy is a rare, progressive muscle disease, presenting in adulthood, mainly affecting proximal limb and bulbar muscles. Muscle biopsies show characteristic nemaline rods. The putative mechanism is considered immune-related. Other manifestations aside from neuromuscular symptoms have not been described previously. We present a case with atypical sporadic late onset nemaline myopathy (SLONM) of a non-HIV, non-MGUS subtype, where skin manifestations preceded neuromuscular symptoms, and a residual thymus with the histology of thymic follicular hyperplasia was detected during the diagnostic workup. Thorough dermatological investigations could not explain the skin presentations. Muscle biopsy revealed variation in fiber diameter, ragged-red and COX-negative fibers associated with discrete fibrosis. Electron microscopy detected atrophic muscle fibres with disorganization of the myofibrils, nemaline rods and abnormal mitochondria. Single-fiber EMG suggested signs of a neuromuscular transmission defect, EMG showed signs of myopathy. Analyses of antibodies associated with myasthenia gravis were negative. The patient showed improvement after intravenous immunoglobulin treatment regarding both the skin and the muscle symptoms. Our case highlights the heterogeneity of SLONM with its varied spectrum of presentation. A unique combination of dermatological symptoms and SLONM could be seen with skin lesions as primary presenting symptoms. An association can be considered between the different manifestations, presumably based on immune etiology, where immunosuppressive therapy has been beneficial.


Esophagectomy with three-field lymph node dissection for esophageal carcinoma with a nonrecurrent inferior laryngeal nerve.
In rare cases, the inferior laryngeal nerve branches directly from the vagus trunk. A 58-year-old man with carcinoma of the thoracic esophagus was referred to our hospital. A nonrecurrent anomaly of the right recurrent laryngeal nerve associated with an aberrant right subclavian artery was detected preoperatively by computed tomography and magnetic resonance imaging. This artery ran on the right side between the esophagus and the vertebral column. Recognition of this nerve before upper mediastinal lymph node dissection was thought to be important for avoiding unexpected neural injuries. For a successful esophagectomy with three-field lymph node dissection in patients associated with this anomaly, a cervico-abdominal procedure followed by a thoracic procedure, which is the reverse of the usual process, could be useful for a safe operation. However, this anomaly made it difficult to dissect lymph node along the left recurrent laryngeal nerve.


The impact of cigarette smoking in predicting stroke using CHADS2 and CHA2DS2-VASc schemas.
To determine the impact of smoking status in the prediction of stroke using CHADS2 and CHA2DS2-VASc schemes. Five hundred twenty-eight consecutive patients with arrhythmic symptoms and without any documented arrhythmia from Queen Mary Hospital, Hong Kong, were followed up to determine the incidence of ischemic stroke, new-onset atrial fibrillation (AF), or all-cause mortality. Smoking status was classified into nonsmokers and smokers. The pairwise comparisons of C-statistics for outcomes were performed. During a median follow-up period of 6.2 years, 65 (12.3%) individuals developed ischemic stroke. Smokers experienced higher annual incidence of stroke, a new-onset AF, and all-cause death compare to nonsmokers, with corresponding hazard ratio (HR) of stroke, AF, and all-cause death being 2.51 (95% confidence intervals, CI 1.36als, CIse death bein 1.15a3.24), and 1.95 (95% CI 1.161.95 (95% CIath being 2.51 (95% confidence corr2 and CHA2DS2-VASc for stroke were 0.60 (95% CI 0.51 for stp = 0.09) and 0.59 (95% CI 0.50 (95%, p = 0.15) respectively, whereas the C-statistics of CHADS2 and CHA2DS2-VASc were 0.66 (95% CI 0.61 were 0p = 0.005), 0.75 (95% CI 0.7 CI 0.7p < 0.0001), respectively among nonsmokers. After incorporating smoking, both the CHADS2-smoking and CHA2DS2-VASc-smoking achieved better C-statistics for new-onset ischemic stroke prediction superior to baseline score systems in male groups. Cigarette smoking status has impact on stroke stratification using CHADS2 and CHA2DS2-VASc scheme. The discrimination of the CHADS2 and CHA2DS2-VASc scheme for stroke can be significantly improved if smoking status is additionally considered.


Interleukin-6 in transcatheter arterial embolization for patients with hepatocellular carcinoma. Effects of serine protease inhibitor.
Modulation of serum levels of circulating cytokines and inflammatory responses with a serine protease inhibitor was studied in 34 patients with hepatocellular carcinoma (HCC) after transcatheter arterial embolization (TAE). The 34 patients were randomly divided into two groups: 17 patients received 500 mg gabexate mesilate, a serine protease inhibitor, intravenously twice a day for 5 days after TAE, and the remaining 17 patients did not receive the drug. In the patients not given the drug, circulating interleukin-6 (IL-6) markedly increased 1 day after TAE, reached a peak (approximately 8 times the pretreatment value) after 4 days, and remained elevated 7 days after TAE. In comparison, in the patients given the drug, circulating IL-6 was at a significantly lower level at 4 and 7 days after TAE (P < 0.05, respectively). Both groups did not show significant change in circulating interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) during the week after TAE. The drug also showed a tendency to keep patient temperature below 38 degrees C, and the elevation of serum C-reactive protein (CRP) concentration to less than 1 mg/dl after TAE (P < 0.05, respectively). The serum level of circulating IL-6 can be modulated by serine protease inhibitor, and this may contribute to suppressing inflammatory responses, such as fever and acute-phase protein synthesis, in the liver after TAE.


Transplantation of immortalized bladder epithelial cell lines in denuded mouse bladder.
The temporal and spatial regeneration of denuded mouse bladders was characterized using antibody markers to mucosal and submucosal elements in bladder tissue. Mechanical stripping of bladder mucosa resulted in a plane of cleavage in the submucosa leaving the muscle layers intact. Fully regenerated bladders were observed after 14 to 21 days although submucosal elements showed abnormal vacuolation. Implantation of immortalized epithelial cell line BBN3 into denuded bladders resulted in solid tumor formation while, in contrast, transplantation of MB331 showed cystic structures with no indication of invasion. Confirmation of the epithelial and implanted origin of cells lining the lumina of implanted bladders were shown using different keratin antibodies. The benign behavior of MB331 in vivo is suggestive of a cell line representing a preneoplastic stage in carcinogenesis and demonstrates an approach to assess the in vivo phenotype of cell parameters established in vitro.


Lanthanides are transported by ionophore A23187 and mimic calcium in the histamine secretion process.
Ionophore A23187 induced histamine release from peritoneal rat mast cells in the presence of lanthanum or terbium as it did in the presence of calcium. Low concentrations of lanthanides (10(-5) to 2 X 10(-4) M) were more efficient than similar concentrations of calcium. The effect of low concentrations of calcium and lanthanides were additive. Increasing the concentration of lanthanides above 10(-3) M decreased histamine release. This decrease was partly reversed by calcium. Calmodulin inhibitors, phenothiazines, R24571 and mepacrine, inhibited the histamine release induced by either calcium or lanthanides. Zn2+, a calmodulin inhibitor transported by A23187, inhibited more potently the calcium-dependent histamine release. Lanthanides decrease histamine release induced by 48/80 in the absence of added calcium. These data show that ionophore A23187 can transport lanthanides across the plasma membrane of mast cells, allowing the trivalent cations to substitute for calcium in the activation of calmodulin or calmoduline-like proteins.


The role of social support in anxiety for persons with COPD.
This study examined the contribution of perceived social support to the presence of anxiety in persons with chronic obstructive pulmonary disease (COPD). A cross-sectional survey sample of 452 persons with COPD (61.3% female; 53.5% older than 65; 70.8% without a college degree or higher educational achievement, and 54.8% with household income of $40,000 or less) completed a telephone survey. Measures included the anxiety subscale of the Hospital Anxiety and Depression Scale (HADS-A), 5 social support subscales from the Positive and Negative Social Exchanges (PANSE) Scale, a COPD Severity Score (CSS; a weighted algorithmic combination of symptoms and the need for various COPD medical interventions), and the Geriatric Depression Scale, Short Form (GDS-SF). Zero order correlations and a series of multiple regression analyses were calculated. Multiple regression analysis revealed that the receipt of instrumental support, feeling let down by the failure of others to provide needed help, and unsympathetic or insensitive behavior from others each positively predicted a higher level of patient anxiety in COPD patients, after controlling for demographic variables, smoking status, comorbid depression (GDS) and severity of illness (CSS). Additionally, the control variable of depression was the strongest predictor of anxiety, suggesting a high degree of co-morbidity in this sample. Anxiety and depression are serious co-morbid mental health concerns for persons with COPD. It is important to examine both positive and negative aspects of perceived social support for COPD patients and how they may impact or interact with these mental health concerns.


Impact of flow, gradient, and left ventricular function on outcomes after transcatheter aortic valve replacement.
To assess the impact of low flow with and without preserved left ventricular ejection fraction (LVEF) on outcomes after transcatheter aortic valve replacement (TAVR). Prior studies have shown that patients with low flow, AVG, and LVEF have worse outcomes after TAVR. It is unclear whether low AVG and LVEF remain prognostic after adjusting for flow, and how the outcomes of patients with low flow with and without preserved LVEF compare after TAVR. The goal of this study was to provide insight into these open questions. Data from 340 TAVR patients at Brigham and Women's Hospital from 2011 through 2015 were analyzed. Low flow was defined as stroke volume index (SVI) ≤35 mL/m2 , low AVG as mean gradient < 40 mmHg, and reduced LVEF as < 50%. Low flow was present in 96 (28.2%) patients, 48 (50.0%) of whom also had reduced LVEF. At 1 year, low flow was associated with increased mortality (21.9 vs 7.4%; P = 0.0002) and heart failure (HF) (20.8 vs 5.3%; P = 0.0113). Among patients with low flow, those with preserved LVEF had increased mortality (HR 5.17, 95% CI 2.73-9.80; P < 0.001) and HF (HR 7.69, 95% CI 3.86-15.31; P < 0.001). After adjusting for clinical factors, patients with low flow had increased mortality (HR 6.51, 95% CI 2.98-14.22; P < 0.001) and HF (HR 5.52, 95% CI 2.34-12.98; P < 0.001), while neither low AVG nor low LVEF were associated with increases in mortality or HF. In patients undergoing TAVR, low flow was an independent predictor of 1-year mortality and HF, and a stronger predictor than either low AVG or LVEF. Patients with low flow and preserved EF had increased mortality and HF at 1-year, while those with low flow and reduced EF had outcomes similar to patients with normal flow.


Endocannabinoid analogues exacerbate marble-burying behavior in mice via TRPV1 receptor.
Activation of cannabinoid CB(1) receptor is shown to inhibit marble-burying behavior (MBB), a behavioral model for assessing obsessive-compulsive disorder (OCD). Anandamide, an endogenous agonist at CB(1) receptor also activates the transient receptor potential vanilloid type 1 (TRPV1) channels but at a higher concentration. Furthermore, anandamide-mediated TRPV1 effects are opposite to that of the CB(1) receptor. Therefore, the present study was carried out to investigate the influence of low and high doses of anandamide on MBB in CB(1) and TRPV1 antagonist pre-treated mice. The results revealed that i.c.v. administration of lower doses of anandamide (1-10 μg/mouse) or its analogues (AM404 or URB597; 1-5 μg/mouse) inhibited MBB indicating the anticompulsive activity. Conversely, at higher doses (40 or 20 μg/mouse) these compounds increased MBB similar to capsaicin (TRPV1 agonist, 100 μg/mouse) exhibiting a pro-compulsive effect. Pretreatment with AM251 (CB(1) antagonist, 1 μg/mouse) antagonized the anticompulsive effect of these compounds, while their pro-compulsive effect at higher doses was attenuated by inactive dose of capsazepine (TRPV1 antagonist, 10 μg/mouse). However, capsazepine per se at a higher dose (100 μg/mouse) inhibited MBB. When given daily for 14 days, the anticompulsive effect of anandamide and its analogues gradually disappeared, whereas capsazepine either alone or with URB597 produced consistent inhibition of MBB comparable to fluoxetine. Thus, the study indicates the biphasic influence of anandamide on MBB, and chronic administration of capsazepine either alone or with URB597 might be an effective tool in the treatment of OCD.


Sagittal craniosynostosis combined with ossified cephalhematoma--a tricky and demanding puzzle.
Four cases of sagittal synostosis combined with ossified cephalhematoma prompted the authors to present the data and to discuss the implications. Large cephalhematoma of the vertex at birth with subsequent ossification occurred in all with final sizes of 6.5-10 by 4.0-5.5 by 0.8-1.8 cm. At surgery with 2-6 months, the mean skull index was 64.75, sagittal suture completely closed, and a disfiguring bony mass present in all. In three of the original cohort of 106 sagittal synostoses, ossified cephalhematoma was removed in one piece together with the suture. In contrast to pathogenesis of common parietal cephalhematomas, cephalhematoma in sagittal synostosis is rather induced by periosteal detachment of the midline by strong shearing forces because molding is hindered in the lateral direction. Frequent and fast complete ossification is possibly directed by the same local factors (e.g., Noggin) which lead to premature fusion of cranial sutures. Some treatment principles of ossified cephalhematoma in sagittal synostosis may be applied to surgery of common types.


Design, Synthesis, and Structure-Activity Relationship Studies of (4-Alkoxyphenyl)glycinamides and Bioisosteric 1,3,4-Oxadiazoles as GPR88 Agonists.
Increasing evidence implicates the orphan G protein-coupled receptor 88 (GPR88) in a number of striatal-associated disorders. In this study, we report the design and synthesis of a series of novel (4-alkoxyphenyl)glycinamides (e.g., 31) and the corresponding 1,3,4-oxadiazole bioisosteres derived from the 2-AMPP scaffold (1) as GPR88 agonists. The 5-amino-1,3,4-oxadiazole derivatives (84, 88-90) had significantly improved potency and lower lipophilicity compared to 2-AMPP. Compound 84 had an EC50 of 59 nM in the GPR88 overexpressing cell-based cAMP assay. In addition, 84 had an EC50 of 942 nM in the [35S]GTPγS binding assay using mouse striatal membranes but was inactive in membranes from GPR88 knockout mice, even at a concentration of 100 μM. In vivo pharmacokinetic testing of 90 in rats revealed that the 5-amino-1,3,4-oxadiazole analogues may have limited brain permeability. Taken together, these results provide the basis for further optimization to develop a suitable agonist to probe GPR88 functions in the brain.


Expression of housekeeping genes during the asexual cell cycle of Plasmodium falciparum.
In an earlier study, we found that calmodulin displayed an atypical expression for a housekeeping gene during the erythrocytic cycle of Plasmodium falciparum. The expression pattern was that of an inducible gene linked to the cell cycle, with a peak prior to replication, and not one of a gene that expresses itself in a constitutive way. In this work, we examined the expression pattern of other housekeeping genes, selecting genes from two functionally very different groups: those for three enzymes involved in carbohydrate metabolism--glucose-phosphate-isomerase (GPI), aldolase and glucose-6-phosphate-dehydrogenase (G6PD)--and for three proteins with structural and motor functions--actin-I, beta-tubulin and myosin. The mRNA of each gene was measured by reverse transcription-polymerase chain reaction in synchronic parasite samples that were 14, 28, 40 and 48 h old. GPI and G6PD achieved their maximum expression at 28 h, then declined, while aldolase increased its expression up to 40 h and remained high, but less so at 48 h. Actin and myosin showed the same pattern, increasing up to 48 h, while beta-tubulin expression peaked at 40 h. These findings confirm unconventional behavior in the expression of certain Plasmodium housekeeping genes and suggest the existence of different expression patterns for distinct functional groups.


Efficacy of loop diuretics in the management of undocumented patients with end-stage renal disease.
An estimated 6000 patients who are undocumented immigrants have end-stage renal disease (ESRD) and routinely present to public safety-net hospitals for life-saving emergent dialysis treatments. Because these patients lack a dialysis unit, they often do not have access to medication management consistently coordinated by a nephrologist, and this can result in more frequent emergency department (ED) utilization and cost of care. We hypothesized that patients who were taking loop diuretics had fewer ED visits for emergency dialysis. Loop diuretics can potentially take advantage of residual renal function and mitigate excess fluid gain that can induce heart failure and high potassium, the two most common indications for emergency dialysis. In our univariable analysis, patients on furosemide had 3.1 fewer ED visits on average compared with patients who are not on furosemide. After adjusting for vintage and serum potassium measures, the average number of ED visits was about 1.1 visits less in furosemide-treated patients compared with patients not receiving furosemide (95% confidence interval, -4.4 to 2.1). These results suggest that loop diuretics may have an important role in undocumented patients with ESRD with residual renal function. Further study to develop practical approaches to the care of undocumented patients with ESRD is greatly needed.


Cell surface and cell division.
A mathematical model of the regulation of cell division is suggested. The model is based on the hypothesis that the process giving rhythm to cell division is located in the cell membrane: i.e., the process of free-radical oxidation of membrane lipids. Much depends on the physical state of the membrane. In the membrane, phase transitions take place because of the changes in lipid composition. These transitions differ in normal and tumor cells: in normal cells they are sharp and hysteretic owing to the presence of a framework (membrane skeleton) on the surface of the membrane, while in tumor cells the integrity of the surface is violated so that the transitions are smooth. This model makes it possible to explain differences in the regulation of normal and cancer cell proliferation. Within the limits of the model, such phenomena as density dependent inhibition of growth, reverse transformation, influence of cyclic AMP and ions of Ca2+ on the cell cycle, the actions of serum and of proteases on the cycle, and so on, are explained. A rational scheme for the appearance of the selective damage found in tumor cells is proposed.


An alternative to interference indexes in neuropsychological time-sharing research.
Interference indexes are used as criterion measures in a significant proportion of time-sharing paradigms in order to control for the initial discrepancy between left- and right-handed tapping rates. Problems associated with these indexes include low reliability, low statistical power, and especially correlations with baseline tapping rates. An analysis of covariance procedure using raw-score concurrent tapping rates as criterion measures, and baseline tapping rates as covariates, can effectively address these problems as demonstrated through a reanalysis of data.


Inoperable chronic thromboembolic pulmonary hypertension treated with riociguat: A case study.
A 51-year-old woman presented with a one-year history of progressive dyspnea, WHO functional class III-IV and exercise-related syncope. Transthoracic echocardiography and computed tomography pulmonary angiography were performed, leading to a diagnosis of pulmonary arterial hypertension. She was referred to our pulmonary hypertension unit, where a complete study was performed, including ventilation/perfusion scan, which was consistent with chronic thromboembolic pulmonary hypertension. Risk factors for this condition were excluded and therapeutic options were evaluated. Imaging studies showed distal pulmonary disease so pulmonary endarterectomy was rejected. Further therapeutic options were evaluated and the patient was subsequently enrolled in an open-label uncontrolled trial with riociguat. After one year of treatment, significant improvement in functional class, 6-minute walk test and NT-proBNP were seen, without significant secondary effects.


Thyroid cancer: the case for total thyroidectomy.
Since there are no prospective studies concerning the treatment of thyroid cancer, there continues to be a considerable disagreement about the 'best' or most appropriate form of surgical treatment for patients with papillary or follicular thyroid cancer. Some surgeons recommend selective treatment depending upon the type of thyroid tumor and stage of the disease. Some advocate thyroid lobectomy and isthmusectomy, some near total thyroidectomy, and some total thyroidectomy for patients with papillary and follicular thyroid cancer. Total thyroidectomy for thyroid cancer would be the treatment of choice for virtually all patients with thyroid cancers if it could be done without complications. We therefore reviewed 160 consecutive patients who had total thyroidectomy for suspected or proven thyroid cancer to determine the complication rate of total thyroidectomy. One hundred and three patients had primary operations, 57 had reoperations with completion of total thyroidectomy and 124 had thyroid cancer. Serious complications (i.e. vocal cord paralysis or hypoparathyroidism) included two cases of transient bilateral recurrent nerve palsy, two patients with presumed transient unilateral vocal cord paralysis, three recurrent laryngeal nerves that were purposely sacrificed because of invasion of the nerve, and one case of permanent hypoparathyroidism. Two other patients developed postoperative wound infections. Only one of the permanent complications, the case of permanent hypoparathyroidism, could have been avoided by a lesser procedure. The experienced surgeon can perform a total thyroidectomy with minimal morbidity, and this procedure has certain theoretical and practical advantages. It should not be done, however, if it will result in a significant complication rate and, in selected patients, it may be preferable to leave a small amount of thyroid tissue to protect the blood supply to the parathyroid glands or recurrent laryngeal nerve.


A cool approach to reducing electrode-induced trauma: Localized therapeutic hypothermia conserves residual hearing in cochlear implantation.
The trauma caused during cochlear implant insertion can lead to cell death and a loss of residual hair cells in the cochlea. Various therapeutic approaches have been studied to prevent cochlear implant-induced residual hearing loss with limited success. In the present study, we show the efficacy of mild to moderate therapeutic hypothermia of 4 to 6 °C applied to the cochlea in reducing residual hearing loss associated with the electrode insertion trauma. Rats were randomly distributed in three groups: control contralateral cochleae, normothermic implanted cochleae and hypothermic implanted cochleae. Localized hypothermia was delivered to the middle turn of the cochlea for 20 min before and after implantation using a custom-designed probe perfused with cooled fluorocarbon. Auditory brainstem responses (ABRs) were recorded to assess the hearing function prior to and post-cochlear implantation at various time points up to 30 days. At the conclusion of the trials, inner ears were harvested for histology and cell count. The approach was extended to cadaver temporal bones to study the potential surgical approach and efficacy of our device. In this case, the hypothermia probe was placed next to the round window niche via the facial recess or a myringotomy. A significant loss of residual hearing was observed in the normothermic implant group. Comparatively, the residual hearing in the cochleae receiving therapeutic hypothermia was significantly conserved. Histology confirmed a significant loss of outer hair cells in normothermic cochleae receiving the surgical trauma when compared to the hypothermia treated group. In human temporal bones, a controlled and effective cooling of the cochlea was achieved using our approach. Collectively, these results suggest that therapeutic hypothermia during cochlear implantation may reduce traumatic effects of electrode insertion and improve conservation of residual hearing.


'Big Data' informed drug development: a case for acceptability.
Data, which help inform various stages of drug product development, are increasingly being collected using newer, more novel platforms, such as mobile applications, and analysed computationally as much larger 'Big Data' data sets, revealing patterns relating to human behaviour and interactions. Medicine acceptability gauges the ability and willingness of patients to take their dosage forms. It has become a crucial human component of drug product design. Vouching for the age appropriateness of medicinal products, acceptability related data are now expected by regulatory bodies. Shifting from traditional paper-based to electronic data-gathering platforms will allow the pharmaceutical industry to collect real-world, real-time, clinically relevant data, capable of informing current and future drug product development, reducing time and cost, and setting foundations for patient-centric drug product design.


Capsid Modifications for Targeting and Improving the Efficacy of AAV Vectors.
In the past decade, recombinant vectors based on a non-pathogenic parvovirus, the adeno-associated virus (AAV), have taken center stage as a gene delivery vehicle for the potential gene therapy for a number of human diseases. To date, the safety of AAV vectors in 176 phase I, II, and III clinical trials and their efficacy in at least eight human diseases are now firmly documented. Despite these remarkable achievements, it has also become abundantly clear that the full potential of first generation AAV vectors composed of naturally occurring capsids is not likely to be realized, since the wild-type AAV did not evolve for the purpose of therapeutic gene delivery. In this article, we provide a brief historical account of the progress that has been made in the development of capsid-modified, next-generation AAV vectors to ensure both the safety and efficacy of these vectors in targeting a wide variety of human diseases.


A new syndrome with ocular, skeletal and renal involvement.
A patient with retinitis pigmentosa, hypertension with interstitial nephropathy, short limb dwarfism with Madelung deformity of the forearms and an unclassified type of brachydactyly is described. Such bone dysplasia has never been reported to date either as a single entity or associated with renal and retinal diseases.


Insulin-like growth factor-1 receptor (IGF-1R) expression does not predict for resistance to trastuzumab-based treatment in patients with Her-2/neu overexpressing metastatic breast cancer.
Her-2/neu and the insulin-like growth factor-1 receptor (IGF-1R) share common postreceptor-signaling pathways, and pre-clinical models have implicated IGF-1R-signaling in resistance to treatment with the anti-Her-2/neu antibody trastuzumab. The present analysis was performed to evaluate the clinical relevance of IGF-1R expression within the context of trastuzumab-based therapy. We performed immunohistochemical (IHC) analysis for IGF-1R expression in tumor specimens from 72 patients receiving trastuzumab-based treatment for Her-2/neu-overexpressing metastatic breast cancer at a single institution. IGF-1R status was evaluated using different cut-offs for positivity regarding staining intensity and staining pattern. IGF-1R positivity was then correlated with clinical patient and biological tumor characteristics and the clinical course of disease of patients under trastuzumab-based therapy. No pattern or intensity of staining for IGF-1R correlated with any of the clinical or biological characteristics. Likewise, response, clinical benefit, progression-free and overall survival were independent of IGF-1R expression in both, univariate and multivariate analyses (all P>0.05). We conclude that IGF-1R expression is not a major predictor of the clinical efficacy of trastuzumab-based treatment in patients with Her-2/neu- overexpressing metastatic breast cancer.


Attenuation of negatively regulated ANP secretion by calcium in hypertrophied atria.
Abnormal intracellular Ca(2+)-handling has been described in various heart diseases associated with cardiac hypertrophy. The crucial role of Ca(2+) in the excitation-secretion coupling in atrial cardiomyocytes is not well established. To investigate modulation of atrial natriuretic peptide (ANP) secretion regulated by Ca(2+) in hypertrophied atria, responsiveness of stretch-induced ANP to Ca(2+) was studied using isolated perfused quiescent hypertrophied rat atria. Male Sprague-Dawley rats were given a single subcutaneous injection of 50 mg/kg monocrotaline (MCT) and were sacrificed at 5-6 weeks. In isolated perfused hypertrophied right atria from MCT rats, changes in atrial volume induced by increased atrial pressure caused proportional increases in mechanically stimulated extracellular fluid (ECF) translocation and stretch-induced ANP secretion. Stretch-induced ANP secretion was markedly increased by the depletion of extracellular Ca(2+). However, an accentuation of stretch-induced ANP secretion by Ca(2+) depletion was markedly attenuated in hypertrophied right atria, as compared to control right atria. Therefore, stretch-induced ANP secretion in terms of ECF translocation by Ca(2+) depletion in hypertrophied atria was significantly lower than in control right atria. However, no significant differences were observed between nonhypertrophied and control left atria. Depletion of extracellular Ca(2+) caused a decrease in intracellular calcium in single beating atrial myocytes, which was significantly attenuated in hypertrophied atrial myocytes. The results suggest that attenuation of Ca(2+)-induced negative regulation of ANP secretion in hypertrophied atria may be due to the disturbance of intracellular Ca(2+) regulation.


Gradual painless visual loss: glaucoma.
Glaucoma is one of the leading causes of blindness in adult Americans. This disease remains undetected in one half of the people with this disease. Primary open angle glaucoma is the most common form of the disease. Patients at risk for glaucoma usually have one or more of the following characteristics: advanced age, diabetes, myopia, elevated intraocular pressure, large cup to disc ratio, and a positive family history of the disease. Presently in the United States, treatment for primary open angle glaucoma is usually medical. Six classes of glaucoma medications are available, all of which have side effects. The second most prevalent form of glaucoma is angle closure glaucoma, the treatment for which is usually surgical. Although a cure for the various forms of glaucoma does not exist, an awareness of the problem and early detection and treatment minimize the impact of the disease.


Nutrient concentrations in a Pampasic first order stream with different land uses in the surrounding plots (Buenos Aires, Argentina).
The objective of this study was to assess the effect of land use on nutrient concentrations in a Pampasic stream. Soluble reactive phosphorus (SRP) concentrations in the stream were higher at a site surrounded by fertilized double-cropped wheat/soybeans than at unfertilized soybeans plots. Nitrate and SRP concentrations in the stream were lower at sites surrounded by soybeans than livestock. It is suggested that crop fertilization and cattle manure increased nutrients loads released to the stream. It is suggested that preservation and restoration of riparian habitats may benefit water quality by decreasing nutrient loads.


Future climates: Markov blankets and active inference in the biosphere.
We formalize the Gaia hypothesis about the Earth climate system using advances in theoretical biology based on the minimization of variational free energy. This amounts to the claim that non-equilibrium steady-state dynamics-that underwrite our climate-depend on the Earth system possessing a Markov blanket. Our formalization rests on how the metabolic rates of the biosphere (understood as Markov blanket's internal states) change with respect to solar radiation at the Earth's surface (i.e. external states), through the changes in greenhouse and albedo effects (i.e. active states) and ocean-driven global temperature changes (i.e. sensory states). Describing the interaction between the metabolic rates and solar radiation as climatic states-in a Markov blanket-amounts to describing the dynamics of the internal states as actively inferring external states. This underwrites climatic non-equilibrium steady-state through free energy minimization and thus a form of planetary autopoiesis.


"Remember this picture when you take more than you need": Constructing morality through instrumental ageism in COVID-19 memes on social media.
As the COVID-19 pandemic continues, a parallel pandemic of ageism is spreading through social media. This paper argues that COVID-19 health securitisation logic and the urgent need to disseminate public health information have allowed nuanced forms of ageism to be reproduced in online forums. I use a critical discourse analysis and social semiotic analysis to deconstruct the use of ageist representations of older adults in COVID-19 memes, which have been organised into four illustrative categories. The analysis attends to ageist representations that both reinforce stereotypical messages and exacerbate intergenerational tensions. Drawing upon moral anthropology, I propose that the memes employ "instrumental ageism," a nuanced form of ageism that advances the health securitisation agenda during the pandemic. The paper concludes with a recognition of the impact of ageist pandemic memes on intergenerational tensions and a call for attention to nuanced forms of ageism in our online and offline social worlds.


Changing the Codon Usage of hfq Gene has Profound Effect on Phenotype and Pathogenicity of Salmonella Typhimurium.
Genome recoding with bias codons (synonymous rare codons) or codon pair bias is being used as a method to attenuate virulence mostly in viruses. The target gene chosen for attenuation in general in bacteria is mostly toxin or virulence gene. We have used RNA chaperone hfq, a global post-transcriptional regulator of bacterial gene expression that regulates about 20 % genes in Salmonella, as the target of recoding. The hfq gene was recoded by replacing the codons of hfq gene with synonymous rare codons. Recoding decreased the expression of Hfq protein about two-fold in the mutant as compared to the parent strain. Recoding did not affect growth kinetics, but in growth competition the mutant strain was outcompeted by the parent strain. There was significant decrease in survivability of mutant strain in macrophage as compared to the parent strain. The biofilm formation was significantly impaired in case of recoded mutant. The mutants were also less motile as compared to the parent strain. Intraperitoneal infection of mice with the mutant strain had shown better survival as compared to parent strain. The results show that recoding is an effective method of reducing virulence.


Membrane protein structural biology in the era of single particle cryo-EM.
In the past few years, significant technological breakthroughs in single particle cryo-electron microscopy enabled a 'resolution revolution' of this technique. It also changed structural biology in an unprecedented way. For many biological macromolecules, obtaining well-ordered crystals of suitable size is no longer a prerequisite for determining their atomic structures. One of the most impacted areas is the structural biology of integral membrane proteins. New structures are now determined at a rapid pace. Despite these advances, further technological developments are still required to overcome new technical challenges that face membrane protein structural biology. In this review, I attempt to discuss some of these challenges.


Thoracic metastases from nasopharyngeal carcinoma presenting as hypertrophic pulmonary osteoarthropathy: scintigraphic and CT findings.
The imaging findings in a group of 33 patients who developed thoracic metastases following prior therapy for nasopharyngeal carcinoma (NPC) were reviewed. Four (12.1%) patients had scintigraphic or radiographic evidence of hypertrophic pulmonary osteoarthropathy (HPOA) on presentation. In one case this developed prior to radiographic or CT evidence of pulmonary metastatic disease. Evidence of hilar or mediastinal metastatic involvement was seen in three of four patient, and a pulmonary metastasis alone in one. Similarity in appearance of the hilar or mediastinal deposits to primary bronchial carcinoma and the presence of HPOA necessitated biopsy confirmation of NPC metastasis in all four cases. The scintigraphic and CT appearances of this unusual radiological association are discussed.


Coronary reserve in Takayasu's arteritis: transesophageal echocardiographic analysis.
The incidence of coronary arteries lesion in Takayasu's arteritis varies from 9% to 10% and is usually discovered at autopsy. Recent studies have demonstrated the value of echocardiography in noninvasive detection of significant coronary stenosis. The aim of our study was to evaluate coronary reserve in 15 patients with Takayasu's arteritis using contrast transesophageal echocardiography (Optison). Transesophageal echocardiogram under basal conditions and in the hyperemic phase with dipyridamole challenge, myocardial perfusion studies and coronary angiography were performed on all patients. Seventy-three percent of them had histories of systemic hypertension, and the most frequent cardiovascular symptoms were shortness of breath (80%), headache (46%), angina (40%), and dizziness (33%). The transthoracic echocardiogram showed left ventricular systolic dysfunction in 6% and diastolic dysfunction in 53%. Aortic regurgitation was found in 67% of the patients, and 60% had mitral and/or tricuspid regurgitation. Coronary reserve was diminished in 3 patients with significant coronary lesions and in 2 patients with coronary dilation (33%). In the 3 patients with obstructive lesions and diminished coronary reserve, reversible perfusion defects were found with thallium-201 single-photon emission computed tomography. One patient with coronary dilation had normal perfusion, while the other demonstrated reversible and irreversible perfusion defects. One patient without coronary lesions and with normal coronary reserve had an irreversible fibrotic changes and the other reversible defects due to abnormal microcirculation. Transesophageal echocardiography is a feasible and promising technique for assessing coronary reserve in patients with Takayasu's arteritis.


Urn sampling and the proportional hazard model.
This paper investigates the urn sampling analogue for the score statistic relating survival to covariates assuming a proportional hazard model. The exact permutation distribution can be calculated as well as the exact low order moments for arbitrary censoring patterns. The asymptotic distribution of the score statistic is an easy consequence. The method is naturally extended to deal with the multivariate case, time varying covariates and interval censoring. Finally the relationship between the censoring process, the survival times and covariates are studied considering different reference sets for the distribution of the score statistic. Some assumptions about the censoring process are investigated and as a consequence the effect of censoring is clarified.


A helical string of alternately connected three-helix bundles for the cell wall-associated adhesion protein Ebh from Staphylococcus aureus.
The 1.1 MDa cell-wall-associated adhesion protein of staphylococci, Ebh, consists of several distinct regions, including a large central region with 52 imperfect repeats of 126 amino acid residues. We investigated the structure of this giant molecule by X-ray crystallography, circular dichroism (CD) spectrometry, and small-angle X-ray scattering (SAXS). The crystal structure of two repeats showed that each repeat consists of two distinct three-helix bundles, and two such repeats are connected along the long axis, resulting in a rod-like structure that is 120 A in length. CD and SAXS analyses of the samples with longer repeats suggested that each repeat has an identical structure, and that such repeats are connected tandemly to form a rod-like structure in solution, the length of which increased proportionately with the number of repeating units. On the basis of these results, it was proposed that Ebh is a 320 nm rod-like molecule with some plasticity at module junctions.


Anti-tumor evaluation of a novel methoxyphenyl substituted chlorin photosensitizer for photodynamic therapy.
Photodynamic therapy (PDT) is a non-invasive and innovative therapeutic approach which has been increasingly applied in clinical cancer therapy. As the central element of PDT, the development of novel photosensitizers (PSs) with longer absorption wavelength, proper lipophilic/hydrophilic profiles, target tissue selectivity, and higher photo-/lowest dark-cytotoxicity is a challenging task. Previously, we designed and synthesized a series of novel long-wavelength chlorin e6 (Ce6)-based PSs via introducing aromatic groups to the vinyl of Ce6 skeleton. The new formed compounds with π-extension system exhibited improved photodynamic effects and spectral characteristics. Among these π-conjugated chlorin PSs, (E)-32-(4-methoxyphenyl)-chlorin e6, named A15, was expected to be a potent antitumor candidate as a PDT agent due to its good photobiological properties. Herein, in this work, we evaluated the effectiveness of A15 in cancer PDT. In vitro, a novel rare earth probe, ATTA-Eu3+ was applied to detect the singlet oxygen (1O2) production of A15 in solution and human hepatoma HepG2 cells, respectively. Moreover, A15 exhibited strong phototoxicity and weak dark cytotoxity to HepG2 cells. In H22 tumor bearing mice, A15 showed excellent tumor accumulation ability via i.v. administration and induced tumor regression, followed by laser treatment. These results indicated that A15 is a potential novel π-extension chlorin-type PS for PDT applications.


Overexpression of GbRLK, a putative receptor-like kinase gene, improved cotton tolerance to Verticillium wilt.
Verticillium dahliae is a causative fungal pathogen and only a few genes have been identified that exhibit critical roles in disease resistance and few has shown positive effects on the resistance to Verticillium wilt in transgenic cotton. We cloned a receptor-like kinase gene (GbRLK) induced by Verticillium dahliae (VD) in the disease-resistant cotton Gossypium barbadense cv. Hai7124. Northern blotting revealed that the GbRLK was induced by VD at 96 h after inoculation. The functional GbRLK is from D subgenome since a single base deletion results in a frameshift or dysfunctional homologue in the A subgenome in tetraploid cotton. To verify the function of GbRLK, we developed the overexpression transgenic GbRLK cotton and Arabidopsis lines, and found that they all showed the higher resistance to Verticillium in the greenhouse and field trial. The results of the expression profile using transgenic and non-transgenic Arabidopsis thaliana revealed that the GbRLK regulated expressions of a series genes associated with biotic and abiotic stresses. Therefore, we propose that the increased resistance to Verticillium dahliae infection in transgnic plants could result from reduction in the damage of water loss and regulation of defense gene expression.


Body mass index percentiles among adolescent girls living in Edirne, Turkey.
Body mass index (BMI) is the simplest way to measure obesity; therefore, it is chosen by many authorities as a screening method for adolescent obesity. Body mass index is positively correlated with the complications of childhood and adolescent obesity, such as hypercholesterolemia, insulin resistance, hyper-tension and long-term development of cardiovascular diseases. The aim of the present study was to produce percentile curves for bodyweight, height and BMI in a representative sample of adolescent girls living in urban and rural areas of Edirne, Turkey, and to compare these percentile curves with curves from other countries. The present study was a cross-sectional study, including a representative sample of 1687 adolescent girls from rural and urban areas of Edirne, who were evaluated between May and July 2001. Bodyweight and height were measured using standard procedures. Body mass index (kg/m2) was calculated as the ratio of bodyweight to body height squared. Smoothed percentiles for these variables were calculated using polynominal regression models. Crude weight, height and BMI percentile values, as well as smoothed percentile curves are presented. Body mass index, weight and height reference curves for adolescent girls were produced. When we compared the BMI values of subjects in the present study with those of other countries, 85th and 95th percentiles of BMI in the present study were found to be generally lower than those for other ethnicities. Our findings show ethnic differences in BMI among adolescent girls. It will be usefull for each country to produce its own BMI percentiles.


Oxidant-mediated activation of mitogen-activated protein kinases and nuclear transcription factors in the cardiovascular system: a brief overview.
In response to oxidant stress, the cardiovascular system is known to express a number of genes, which could occur owing to the participation of mitogen-activated protein kinases such as MAPKs, ERK and JNK (SAPK) followed by stimulation of at least two well-defined transcription factors NF-KB and AP-1 (c-Fos and c-Jun). Oxidants activate cytosolic and membrane-bound PLA2 activities with the subsequent production of AA metabolites such as HETEs, which subsequently stimulate ERK and JNK (SAPK) activities leading to the activation of transcriptional factors and the ultimate stimulation of the transcription of several mitogen-stress-responsive genes. LacCer, a ceramide analogue present in atherosclerotic plaques, has been found to induce proliferation of aortic smooth muscle cells. LacCer is involved in Ras-GTP loading, activation of kinase cascades (MEK, Raf, p44 MAPK) and c-fos expression. TNF-alpha, on the other hand, induces c-fos, c-myc and c-jun expression. Recent investigations link ceramide and its analogues to the extracellular signal-regulated kinase (ERK) cascade, stress-activated protein kinase-c-Jun kinase (SAPK/JNK) cascade and apoptotic responses. These critical steps in the signalling pathways are sensitive to intracellular thiol-redox and protease(s)-antiprotease(s) status, both of which can be modified by oxidants. Because mobilisation of intracellular Ca2+ caused by a variety of signals also plays a role in the activation of the signalling pathways, an important aspect of future work will be to ascertain the roles of oxidants and Ca2+ individually and in combination in the activation of the signalling pathways. The following two important questions also deserve future attention: (1) How does NF-kB shield cells from apoptotic death? and (2) By what mechanisms does the activated NF-kB cause cellular transformation? Furthermore, the role of AP-1 acting as transcriptional activator seems clear, but the target genes remain to be defined.


The limiting rate of the ATP-mediated dissociation of actin from rabbit skeletal muscle myosin subfragment 1.
The ATP-induced dissociation of actoS1 has been studied at temperatures between -10 degrees C and +30 degrees C in a stopped-flow apparatus using ethylene glycol as antifreeze. At temperatures at and below 0 degrees C the observed rate of the dissociation of actin shows a hyperbolic dependence on ATP concentration. This is interpreted in terms of a rapid binding of ATP followed by an isomerisation of the ternary complex which results in actin dissociation. Ethylene glycol weakens ATP binding but the rate of the isomerisation is unaffected. The second order rate constant for the dissociation shows a break in the Arrhenius plot.


Comprehensive Genome profile testing in head and neck cancer.
Head and neck cancer (HNC) is a tumor occurring in various primary sites with limited chemotherapy options for its treatment. Recently, comprehensive genomic profiling (CGP) testing has become clinically widespread. In this study, we examined the utility of CGP in diagnosing and treating HNC. This study included 29 patients with HNC who underwent CGP testing at the Gifu University Hospital between December 2019 and April 2022. We analyzed the types of gene mutations and tumor mutational burden (TMB) based on the CGP results. Squamous cell carcinoma accounted for 55.2%, and other cancers accounted for 44.8%. And we investigated the correlation of prognosis with gene mutations and TMB. Gene mutations were detected in TP53(48.3%), CDKN2A (27.6%), CDKN2B (17.2%), NOTCH1 (17.2%), PIK3CA (17.2%), ARID1A (13.8%), and NF1 (13.8%). TP53, CDKN2A and CDKN2B mutations significantly decreased survival rate in HNC. Five cases (17.2%) were TMB-high and 82.8% were TMB-low. In SCC cases treated with immune checkpoint inhibitors, TMB-high had better Overall survival than TMB-low. And all patients with TMB-high were oropharyngeal cancer. Although there were no cases in which effective treatment was actually performed based on the results of CGP, many gene mutations have been detected and several gene mutations correlated with prognosis. Furthermore, TMB can be used as a biomarker to predict the therapeutic effects of immune checkpoint inhibitors in cases of SCC.


Anti-aggregation-based spectrometric detection of Hg(II) at physiological pH using gold nanorods.
An efficient detection method for Hg (II) ions at physiological pH (pH7.4) was developed using tween 20-modified gold nanorods (NRs) in the presence of dithiothreitol (DTT). Thiol groups (-SH) at the end of DTT have a higher affinity towards gold atoms, and they can covalently interact with gold NRs and leads to their aggregation. The addition of Hg(II) ions prevents the aggregation of gold NRs due to the covalent bond formation between the -SH group of DTT and Hg(II) ions in the buffer system. The changes in the longitudinal surface plasmon resonance peak of gold NRs were characterized using a UV-visible spectrophotometer. The absorption intensity peak of gold NRs at 679nm was observed to reduce after interaction with DTT, and the absorption intensity was noted to increase by increasing the concentration of Hg(II) ions. The TEM analysis confirms the morphological changes of gold NRs before and after addition of Hg(II) ions in the presence of DTT. Further, the aggregation and disaggregation of gold NRs were confirmed by particle size and zeta potential analysis. The developed method shows an excellent linearity (y=0.001x+0.794) for the graph plotted between the absorption ratio and Hg(II) concentration (1 to 100pM) under the optimized conditions. The limit of detection was noted to be 0.42pM in the buffer system. The developed method was tested in simulated body fluid, and it was found to have a good recovery rate.


Predictions of fate from rosette size in four "biennial" plant species: Verbascum thapsus, Oenothera biennis, Daucus carota, and Tragopogon dubius.
Individual plants were marked in field populations of four biennial plant species, Verbascum thapsus L., Oenothera biennis L., Daucus carota L., and Tragopogon dubius Scop., and followed for 2 or 3 years. The relationship of both rosette size and age to the probability of an individual dying, remaining vegetative, or flowering was determined for each species. In all four species, a minimum size must be reached before flowering can be induced and above that minimum the probability of flowering increases directly with rosette size. Conversely, the probability of dying decreases with increasing rosette size. In general, the fate of a rosette appears to be independent of its age although in populations where the probability of death is high for individuals above the minimum size for flowering, the age of a rosette may influence its fate to some extent.


Travel times to hospitals in Australia.
Travel time to hospital is a key measure of health service accessibility, and impacts patients' experiences of care and health outcomes. Methods used to estimate travel time vary across studies. In Australia the smallest geographical areas defined by the Australian Bureau of Statistics for the release of population counts are mesh blocks (MBs) and the smallest geographical areas for the release of health-related statistics are statistical areas level 2 (SA2). SA2s are built up from whole MBs. This project used the Open Source Routing Machine (OSRM) HTTP server to compute estimated travel times between the centroid of each inhabited MB and each hospital in Australia, as well as the shortest travel times between MBs and any hospital. By computing population-weighted averages across MBs, the average travel times to hospitals and the shortest travel time to any hospital were estimated for each SA2. This dataset will promote consistency across studies investigating geographic influences on health care in Australia, and the methods are applicable to generating similar datasets for other countries.


A psychosocial analysis of parents' decisions for limiting their young child's screen time: An examination of attitudes, social norms and roles, and control perceptions.
Preschool-aged children spend substantial amounts of time engaged in screen-based activities. As parents have considerable control over their child's health behaviours during the younger years, it is important to understand those influences that guide parents' decisions about their child's screen time behaviours. A prospective design with two waves of data collection, 1 week apart, was adopted. Parents (n = 207) completed a Theory of Planned Behaviour (TPB)-based questionnaire, with the addition of parental role construction (i.e., parents' expectations and beliefs of responsibility for their child's behaviour) and past behaviour. A number of underlying beliefs identified in a prior pilot study were also assessed. The model explained 77% (with past behaviour accounting for 5%) of the variance in intention and 50% (with past behaviour accounting for 3%) of the variance in parental decisions to limit child screen time. Attitude, subjective norms, perceived behavioural control, parental role construction, and past behaviour predicted intentions, and intentions and past behaviour predicted follow-up behaviour. Underlying screen time beliefs (e.g., increased parental distress, pressure from friends, inconvenience) were also identified as guiding parents' decisions. Results support the TPB and highlight the importance of beliefs for understanding parental decisions for children's screen time behaviours, as well as the addition of parental role construction. This formative research provides necessary depth of understanding of sedentary lifestyle behaviours in young children which can be adopted in future interventions to test the efficacy of the TPB mechanisms in changing parental behaviour for their child's health. What is already known on this subject? Identifying determinants of child screen time behaviour is vital to the health of young people. Social-cognitive and parental role constructions are key influences of parental decision-making. Little is known about the processes guiding parents' decisions to limit their child's screen time. What does this study add? Parental role construction and TPB social-cognitive factors influence parental decisions. The beliefs of parents for their child's behaviour were identified. A range of beliefs guide parents' decisions for their child's screen time viewing.


Odor emission assessment of different WWTPs with Extended Aeration Activated Sludge and Rotating Biological Contactor technologies in the province of Cordoba (Spain).
In this study, five urban WWTPs (Wastewater Treatment Plant) with different biological treatment (Extended Aeration Activated Sludge - EAAS; Rotating Biological Contactor - RBC), wastewater type (Urban; Industrial) and size, were jointly evaluated. The aim was twofold: (1) to analyze and compare their odor emissions, and (2) to identify the main causes of its generation from the relationships between physico-chemical, respirometric and olfactometric variables. The results showed that facilities with EAAS technology were more efficient than RBC, with elimination yields of organic matter higher than 90%. In olfactometric terms, sludge managements facilities (SMFs) were found to be the critical odor source in all WWTPs compared to the Inlet point (I) or Post primary treatment (PP), and for seasonal periods with ambient temperature higher than 25 °C. Moreover, the global odor emissions quantified in all SMFs revealed that facilities with EAAS (C-WWTP, V-WWTP and Z-WWTP) had a lower odor contribution (19,345, 14,800 and 11,029 ouE/s·m2, respectively) than for those with RBC technology (P-WWTP and NC-WWTP) which accounted for 19,747 ouE/s·m2 and 80,061 ouE/s·m2, respectively. In addition, chemometric analysis helped to find groupings and differences between the WWTPs considering the wastewater (71.27% of total variance explained) and sludge management (64.52% of total variance explained) lines independently. Finally, odor emissions were adequately predicted from the physico-chemical and respirometric variables in the wastewater (r2 = 0.8738) and sludge (r2 = 0.9373) lines, being pH, volatile acidity and temperature (wastewater line), and pH, moisture, temperature, SOUR (Specific Oxygen Uptake Rate) and OD20 (Cumulative Oxygen Demand at 20 h) (sludge line) the most influential variables.


Microbial diversity in natural whey cultures used for the production of Caciocavallo Silano PDO cheese.
The microbial diversity of sixty-three Natural Whey Cultures (NWCs) for the manufacture of Caciocavallo Silano cheese PDO was studied. The NWCs were collected from different dairies covering the whole territory of PDO production including five different regions of southern Italy. The microbial species diversity was determined by direct DNA extraction from NWCs and Polymerase Chain Reaction (PCR) amplification of variable regions of the 16S rRNA gene followed by denaturing gradient gel electrophoresis (DGGE) and denaturing high performance liquid chromatography (DHPLC). DGGE and DHPLC fingerprinting yielded the same results in terms of number of bands/peaks and specific migration/retention time of the amplicons. The DHPLC technique was used in this study for the first time to assess a food-related mixed microbial community by a culture-independent approach and proved to be at least as effective as DGGE in profiling species diversity in NWCs. Cluster analysis of DGGE and DHPLC data revealed that the species-related groups of similarity were not dependent on the geographical origin of the NWCs. The presence of three groups of 10-14 NWCs at 100% of species similarity indicated that some species associations are very commonly occurring in the NWCs for Caciocavallo Silano cheese PDO. A RAPD-PCR analysis of the NWCs was also performed for the members of the above groups and it showed that, though characterized by the same species diversity, most of the identical NWCs included different biotypes. The sequences of DGGE bands and DHPLC peaks revealed the occurrence of mainly thermophilic lactic acid bacteria such as Lactobacillus delbrueckii, L. helveticus and Streptococcus thermophilus even though the mesophilic Lactococcus lactis also occurred in some NWCs. In conclusion, the results of this study indicate that the microbial diversity of NWCs used for the Caciocavallo Silano PDO cheese is not high, it is not dependent on the geographical origin and the same microbial species occur within the territory examined. The microbiota of fermented PDO products and its possible link with territory should be studied case by case in order to have useful evidences for the assessment of product quality and authenticity.


Generation of a CHIP isogenic human iPSC-derived cortical neuron model for functional proteomics.
The neuroprotective E3-ubiquitin ligase CHIP is linked to healthy aging. Here, we present a protocol using a patient-derived iPSC line with a triplication of the α-synuclein gene to produce gene-edited cells isogenic for CHIP. We describe iPSC differentiation into cortical neurons and their identity validation. We then detail mass spectrometry-based approaches (SWATH-MS) to identify dominant changes in the steady state proteome generated by loss of CHIP function. This protocol can be adapted to other proteins that impact proteostasis in neurons. For complete details on the use and execution of this protocol, please refer to Dias et al. (2021).


Clinical pharmacokinetics of urapidil.
Urapidil is a selective alpha 1-adrenoceptor antagonist with central antihypertensive action which is increasingly used in the treatment of hypertension. Urapidil is readily absorbed, is subject to moderate first-pass metabolism and is eliminated primarily as metabolites of much lower antihypertensive activity than the parent drug. The influences of age, renal and hepatic disease on the disposition of urapidil are reviewed. Studies on the relationship between pharmacodynamics and pharmacokinetics show that the optimum use of urapidil in clinical practice depends on an understanding of the pharmacokinetic properties of the drug.