Prevention of exacerbations of chronic obstructive pulmonary disease with tiotropium, a once-daily inhaled anticholinergic bronchodilator: a randomized trial.
Patients with chronic obstructive pulmonary disease (COPD) frequently develop exacerbations, leading to major clinical and health resource use ramifications. To prospectively evaluate the effectiveness of a long-acting inhaled anticholinergic bronchodilator, tiotropium, in reducing COPD exacerbations and exacerbation-related health care utilization. Randomized, double-blind study. 26 Veterans Affairs medical centers. 1829 patients with moderate to severe COPD (mean baseline FEV(1), 36% predicted). Once-daily tiotropium (18 microg) or placebo for 6 months. Patients otherwise received usual care, except for other anticholinergic bronchodilators. The coprimary end points were the percentage of patients with a COPD exacerbation and the percentage of patients with a COPD-related hospitalization. Tiotropium significantly reduced the percentage of patients experiencing 1 or more exacerbations compared with placebo (27.9% vs. 32.3%, respectively; difference, -5.7 percentage points [95% CI, -10.4 to -1.0 percentage points]; P = 0.037). Fewer tiotropium patients were hospitalized because of COPD exacerbation (7.0% vs. 9.5%, respectively; difference, -3.0 percentage points [CI, -5.9 to -0.1 percentage points]; P = 0.056), although this difference was of borderline statistical significance. Analysis of secondary outcomes indicates that tiotropium may lengthen the time to first COPD exacerbation (P = 0.028) and reduce health care utilization for exacerbations, including the frequency of hospitalizations (P = 0.047), unscheduled clinic visits (P = 0.019), and days of antibiotic treatment (P = 0.015). Tiotropium did not statistically significantly reduce all-cause hospitalization rates. Trial participants were enrolled from 1 health care system, and 99% were men. The follow-up period extended for only 6 months. Tiotropium reduces COPD exacerbations and may reduce related health care utilization in patients with moderate to severe COPD.


The tegmental proliferation region in the sea lamprey.
The diencephalic/midbrain tegmental domain of the developing lamprey was characterized by the special features of the ventricular zone and distribution of some postmitotic neuronal populations, using proliferating cell nuclear antigen (PCNA) and HNK-1 immunocytochemistry. In late prolarvae and early larvae, the tegmental ventricular zone differentiated a striking arrangement of thin longitudinal crests with strongly PCNA-immunoreactive cells protruding toward the ventricle and separated by shallow valleys whose cells were faintly or moderately PCNA-immunoreactive. The tegmental ventricular zone was composed of a large caudal region located ventral to the pretectum and the midbrain tectum, and of a smaller rostral wedge-shaped region that extended dorsally between the dorsal and the ventral thalamus, in the last one, ventricular crests coursing in the zona limitans intrathalamica. In other diencephalic and midbrain regions, the ventricular zone lacked this crest-like organization and exhibited different smooth-surfaced domains rather homogeneous in thickness and appearance. The distribution of HNK-1-immunoreactive cell populations in the tegmental domain appears to be closely related with the pattern of proliferation crests. Together, our results reveal a shared basic organization in the tegmental domains of the diencephalon and midbrain of developing lamprey, indicating early appearance of the domain in vertebrate phylogeny.


Precise in vivo functional analysis of DNA variants with base editing using ACEofBASEs target prediction.
Single nucleotide variants (SNVs) are prevalent genetic factors shaping individual trait profiles and disease susceptibility. The recent development and optimizations of base editors, rubber and pencil genome editing tools now promise to enable direct functional assessment of SNVs in model organisms. However, the lack of bioinformatic tools aiding target prediction limits the application of base editing in vivo. Here, we provide a framework for adenine and cytosine base editing in medaka (Oryzias latipes) and zebrafish (Danio rerio), ideal for scalable validation studies. We developed an online base editing tool ACEofBASEs (a careful evaluation of base-edits), to facilitate decision-making by streamlining sgRNA design and performing off-target evaluation. We used state-of-the-art adenine (ABE) and cytosine base editors (CBE) in medaka and zebrafish to edit eye pigmentation genes and transgenic GFP function with high efficiencies. Base editing in the genes encoding troponin T and the potassium channel ERG faithfully recreated known cardiac phenotypes. Deep-sequencing of alleles revealed the abundance of intended edits in comparison to low levels of insertion or deletion (indel) events for ABE8e and evoBE4max. We finally validated missense mutations in novel candidate genes of congenital heart disease (CHD) dapk3, ube2b, usp44, and ptpn11 in F0 and F1 for a subset of these target genes with genotype-phenotype correlation. This base editing framework applies to a wide range of SNV-susceptible traits accessible in fish, facilitating straight-forward candidate validation and prioritization for detailed mechanistic downstream studies.


What is normal sleep in the elderly?
Changes in the sleep and daytime alertness of the elderly are common and are secondary to a variety of causes. These changes cannot be attributed solely to the aging process. The role of occult sleep disorders in producing these changes in significant, and severe sleep disruption in the healthy elderly is almost always secondary to a sleep disorder such as sleep apnea. A number of precautions must be kept in mind when interpreting studies of sleep in the elderly.


Venous invasion in renal cell carcinoma: a correlative clinical and radiologic study.
Angiography, cavography, ultrasound (US), computed tomography (CT), and low-field magnetic resonance imaging (MRI) were used to explore venous tumor invasion in 86 patients with renal cell carcinoma. The findings confirm previous experience that ultrasound and dynamic CT are reliable methods for the evaluation of venous invasion. Low-field magnetic resonance imaging does not improve tumor staging. In case of equivocal findings, vena cavography should be used as a complementary examination.


Sequence-based HLA-A, B, C, DP, DQ, and DR typing of 100 Luo infants from the Boro area of Nyanza Province, Kenya.
One hundred healthy infants enrolled as controls in a tuberculosis vaccine study in Nyanza Province, Kenya provided anonymized samples for DNA sequence-based typing at the HLA-A, -B, -C, -DPB1, -DQA1, -DQB1, -DRB1, and -DRB3/4/5 loci. The purpose of the study was to characterize allele frequencies in the local population, to support studies of T cell immunity against pathogens, including Mycobacterium tuberculosis. There are no detectable deviations from Hardy Weinberg proportions for the HLA-B, -C, -DRB1, -DPB1, -DQA1 and -DQB1 loci. A minor deviation was detected at the HLA-A locus due to an excess of HLA-A*02:02, 29:02, 30:02, and 68:02 homozygotes. The genotype data are available in the Allele Frequencies Net Database under identifier 3393.


MSGANet-RAV: A multiscale guided attention network for artery-vein segmentation and classification from optic disc and retinal images.
Retinal and optic disc images are used to assess changes in the retinal vasculature. These can be changes associated with diseases such as diabetic retinopathy and glaucoma or induced using ophthalmodynamometry to measure arterial and venous pressure. Key steps toward automating the assessment of these changes are the segmentation and classification of the veins and arteries. However, such segmentation and classification are still required to be manually labelled by experts. Such automated labelling is challenging because of the complex morphology, anatomical variations, alterations due to disease and scarcity of labelled data for algorithm development. We present a deep machine learning solution called the multiscale guided attention network for retinal artery and vein segmentation and classification (MSGANet-RAV). MSGANet-RAV was developed and tested on 383 colour clinical optic disc images from LEI-CENTRAL, constructed in-house and 40 colour fundus images from the AV-DRIVE public dataset. The datasets have a mean optic disc occupancy per image of 60.6% and 2.18%, respectively. MSGANet-RAV is a U-shaped encoder-decoder network, where the encoder extracts multiscale features, and the decoder includes a sequence of self-attention modules. The self-attention modules explore, guide and incorporate vessel-specific structural and contextual feature information to segment and classify central optic disc and retinal vessel pixels. MSGANet-RAV achieved a pixel classification accuracy of 93.15%, sensitivity of 92.19%, and specificity of 94.13% on LEI-CENTRAL, outperforming several reference models. It similarly performed highly on AV-DRIVE with an accuracy, sensitivity and specificity of 95.48%, 93.59% and 97.27%, respectively. The results show the efficacy of MSGANet-RAV for identifying central optic disc and retinal arteries and veins. The method can be used in automated systems designed to assess vascular changes in retinal and optic disc images quantitatively.


Mitochondrial abnormalities in temporal lobe of autistic brain.
Autism spectrum disorder (ASD) consists of a group of complex developmental disabilities characterized by impaired social interactions, deficits in communication and repetitive behavior. Multiple lines of evidence implicate mitochondrial dysfunction in ASD. In postmortem BA21 temporal cortex, a region that exhibits synaptic pathology in ASD, we found that compared to controls, ASD patients exhibited altered protein levels of mitochondria respiratory chain protein complexes, decreased Complex I and IV activities, decreased mitochondrial antioxidant enzyme SOD2, and greater oxidative DNA damage. Mitochondrial membrane mass was higher in ASD brain, as indicated by higher protein levels of mitochondrial membrane proteins Tom20, Tim23 and porin. No differences were observed in either mitochondrial DNA or levels of the mitochondrial gene transcription factor TFAM or cofactor PGC1α, indicating that a mechanism other than alterations in mitochondrial genome or mitochondrial biogenesis underlies these mitochondrial abnormalities. We further identified higher levels of the mitochondrial fission proteins (Fis1 and Drp1) and decreased levels of the fusion proteins (Mfn1, Mfn2 and Opa1) in ASD patients, indicating altered mitochondrial dynamics in ASD brain. Many of these changes were evident in cortical pyramidal neurons, and were observed in ASD children but were less pronounced or absent in adult patients. Together, these findings provide evidence that mitochondrial function and intracellular redox status are compromised in pyramidal neurons in ASD brain and that mitochondrial dysfunction occurs during early childhood when ASD symptoms appear.


Selective cytotoxic actions of aspirin on parietal cells: a principal factor in the early stages of aspirin-induced gastric damage.
Electronmicroscopic observations are reported on the effects of orally administered aspirin on the rat gastric mucosa to establish the cytotoxic events that occur during the early stages of the development of gastric damage induced by this drug. The results show that aspirin, apart from damaging superficial mucous cells, causes selective damage to the acid-secreting parietal cells located deep in the fundic mucosa. It is suggested that the selective accumulation of salicylates in the parietal cells may account for the specific cytotoxic actions of aspirin on these cells.


Insect protein digestion improves purity of Steinernema carpocapsae in vitro culture and reduces culture period.
Insect protein, used for in vitro culture media for entomopathogenic nematode, produces nematodes of high quality. However, the time-consuming culture and poor purity of nematodes hinder the commercial application of insect protein media. We show that hydrolyzed insect protein improves nematode purity in in vitro culture. The results revealed that nematode purity was increased by more than 90 %, and the culture period was reduced by 6 days. Estimated economic efficiency of using hydrolyzed insect protein medium was increased by 44.25 % over that obtained with non-hydrolyzed insect medium.


Identifying the effective behaviour change techniques in nutrition and physical activity interventions for the treatment of overweight/obesity in post-treatment breast cancer survivors: a systematic review.
Updated evidence for the treatment of obesity in cancer survivors includes behavioural lifestyle interventions underpinning at least one theoretical framework. The aim of this systematic review was to assess the effectiveness of theory-based lifestyle interventions for the treatment of overweight/obesity in breast cancer survivors and to report effective behavioural change techniques (BCTs) and components used in these interventions. Four databases were searched for RCTs published between database inception and July 2022. The search strategy included MeSH terms and text words, using the PICO-framework to guide the eligibility criteria. The PRISMA guidelines were followed. Risk-of-bias, TIDier Checklist for interventions' content, and the extent of behaviour change theories and techniques application were assessed. To evaluate the effectiveness of interventions, trials were categorised as "very," "quite," or "non" promising according to their potential to reduce body weight, and BCTs promise ratios were calculated to assess the potential of BCTs within interventions to decrease body weight. Eleven RCTs met the inclusion criteria. Seven trials were classified as "very", three as "quite" and one study was "non" promising. Studies' size, design, and intervention strategies varied greatly, but the weight-loss goal in all studies was ≥ 5% of the initial body weight through a 500-1000 kcal/day energy deficit and a gradually increased exercise goal of ≥ 30 min/day. Social Cognitive Theory was the most commonly used theory (n = 10). BCTs ranged from 10 to 23 in the interventions, but all trials included behaviour goal setting, self-monitoring, instructions on the behaviour, and credible source. The risk-of-bias was "moderate" in eight studies and "high" in three. The present systematic review identified the components of theory-based nutrition and physical activity behaviour change interventions that may be beneficial for the treatment of overweight/obesity in breast cancer survivors. The strategies mentioned, in addition to reported behavioural models and BCTs, should be considered when developing weight-loss interventions for breast cancer survivors.


Texture analysis on MRI images of non-Hodgkin lymphoma.
The aim here is to show that texture parameters of magnetic resonance imaging (MRI) data changes in lymphoma tissue during chemotherapy. Ten patients having non-Hodgkin lymphoma masses in the abdomen were imaged for chemotherapy response evaluation three consecutive times. The analysis was performed with MaZda texture analysis (TA) application. The best discrimination in lymphoma MRI texture was obtained within T2-weighted images between the pre-treatment and the second response evaluation stage. TA proved to be a promising quantitative means of representing lymphoma tissue changes during medication follow-up.


Is elevated plasma B-natriuretic peptide in amyloidosis simply a function of the presence of heart failure?
This study sought to determine plasma levels of B-natriuretic peptide (BNP) in patients with light-chain-associated amyloidosis and correlate them with the presence or absence of heart failure (HF) and the presence or absence of echocardiographic abnormalities. Patients with normal echocardiographic results had significantly lower BNP levels than those with echocardiographic features of cardiac amyloidosis, whereas BNP levels in the group with HF did not differ from those in patients with asymptomatic cardiac amyloidosis. This observation supports previous observations, suggesting that the elevation of BNP in cardiac amyloidosis may be due not only to elevated ventricular filling pressure but also to direct myocyte damage due to extracellular deposits of amyloid.


A 28 kDa protein of normal mouse serum binds lipopolysaccharides of gram-negative and lipoteichoic acids of gram-positive bacteria.
A 28 kDa protein from normal mouse serum known to bind to the inner core region of bacterial lipopolysaccharide (LPS) was found to bind also to bacterial poly(glycerophosphate) lipoteichoic acid (LTA). Twenty-nine preparations of LTA were isolated from 19 different bacterial species, purified, chemically analysed, and tested for their ability to bind the 28 kDa protein in a complement-dependent hemolysis and hemolysis inhibition assay. All but one were active in one or both systems and one half of the preparations were active in both. Reactivity patterns were not strictly correlated with the chemical structure of LTA considering the substitution of the poly(glycerophosphate) chain with alanine ester and glycosyl residues and the type of lipid anchor. The isolated lipid anchor alone was unable to bind the serum factor. Comparing the binding to LTA and LPS from Acinetobacter calcoaceticus indicated complete cross-reactivity of LTA and LPS in various serological approaches. Thus, LPS and LTA which are unique amphiphiles in Gram-negative and Gram-positive bacteria, respectively, share a similar function in terms of binding the 28 kDa mouse serum protein.


Nitric oxide synthase and NADPH-diaphorase after acute hypobaric hypoxia in the rat caudate putamen.
Changes in the production system of nitric oxide (NO), a multifunctional biological messenger known to participate in blood-flow regulation, neuromodulation, and neuroprotection or neurotoxicity, were investigated in the caudate putamen of adult rats submitted to hypobaric hypoxia. Employing immunohistochemistry, Western blotting, enzymatic assay, and NADPH-diaphorase staining, we demonstrate that neuronal nitric oxide synthase (nNOS) expression and constitutive nitric oxide synthase (cNOS) activity were transiently activated by 7 h of exposure to a simulated altitude of 8325 m (27,000 ft). In addition, endothelial nitric oxide synthase (eNOS) immunoreactivity and blood vessel NADPH-diaphorase staining peaked immediately after the hypoxic stimulus, whereas inducible nitric oxide synthase (iNOS) expression and activity remained unaltered. Nitrotyrosine formation, a marker of protein nitration, was evaluated by immunohistochemistry and Western blotting, and was found to increase parallel to nitric oxide synthesis. We conclude that the nitric oxide system undergoes significant transient alterations in the caudate putamen of adult rats submitted to acute hypobaric hypoxia.


DNA-protein cross-linking between thymine and tyrosine in chromatin of gamma-irradiated or H2O2-treated cultured human cells.
Formation of DNA-protein cross-links between thymine and tyrosine in chromatin of gamma-irradiated or H2O2-treated cultured human cells is reported. Chromatin was isolated from cells, and subsequently hydrolyzed and derivatized. Analysis of derivatized hydrolysates by gas chromatography/mass spectrometry with selected-ion monitoring showed that 3-[(1,3-dihydro-2,4-dioxopyrimidin-5-yl)-methyl]-L-tyrosine (Thy-Tyr cross-link) was formed. The presence of this DNA-protein cross-link in control cells was also observed at a level of approximately 7 molecules per 10(6) DNA nucleotides. Exposure of cells to ionizing radiation at doses between 8.7 and 82 Gy (J.kg-1) increased the amount of the Thy-Tyr cross-link linearly up to approximately fourfold over the background level. At doses higher than 82 Gy, the yield approached a plateau. Treatment of cells with H2O2 (0.5 to 10 mM) also increased the amount of the Thy-Tyr cross-link in a concentration-dependent manner. Addition of dimethyl sulfoxide and o-phenanthroline in the culture medium afforded partial inhibition of cross-link formation. Addition of catalase inhibitor KCN prior to H2O2 treatment increased the yield of cross-linking over the level observed with H2O2 treatment alone. Pretreatment of cells with ascorbic acid for 24 h without H2O2 caused formation of the Thy-Tyr cross-link. This DNA-protein cross-link in chromatin of cells is proposed to be formed by mechanisms involving a radical addition reaction and/or a radical-radical combination involving thymine and tyrosine radicals. Hydroxyl radical mediated by chromatin-bound metal ions is proposed to cause the formation of the Thy-Tyr cross-link in H2O2-treated cells.


A nomogram for planning respiratory therapy.
A nomogram is presented which aids in monitoring and selecting therapy in the treatment of the patient with hypoxemia. It facilitates accurate bedside determination of total shunt fraction and estimation of the arterial oxygen tension (PaO2) attainable at inspired oxygen concentrations of 21 percent to 100 percent. The nomogram permits rapid consideration of changes in hemoglobin concentration, arteriovenous oxygen content difference, and shunt fraction on the PaO2. The uses of the nomogram in several therapeutic contexts are discussed.


Developing Educational Health Modules to Improve Vocal Wellness in Mask-Wearing Occupational Voice Users.
To develop educational modules to improve vocal wellness and optimize communication in mask-wearing occupational voice users. Module development focused on identifying accurate, understandable, and actionable steps to improve vocal wellness in the workplace. We i) interviewed eight voice-specialized speech-language pathologists and researchers on current speech and voice recommendations for mask-wearers, ii) developed educational content using the standardized Patient Education Materials Assessment Tool (PEMAT), iii) assessed the ability of nine mask-wearing community members to learn educational content, and iv) compared behavioral, acoustical, and perceptual changes in four mask-wearing healthcare professionals following educational training. We created three educational modules that described key vocal health and communication strategies, including microphone amplification, postural alignment, clear speech, hydration, vocal naps, and vocal warm-ups. PEMAT scores were 96% and 93% on understandability and actionability, respectively. Mask-wearing healthcare professionals increased use of 4 out of the 6 strategies following educational training and were able to retain information at rates >90% at 1-week follow-up. We developed a set of free-to-use educational modules to promote vocal wellness among mask-wearing occupational voice users (see VSMechLab.com). Future work should examine the impact of these strategies on voice measures in a larger group of mask-wearing community members.


Progesterone therapy in women with complex partial and secondary generalized seizures.
This open trial assessed the effects of adjunctive progesterone therapy on seizure frequency in 25 women with catamenial exacerbation of complex partial (CPS) and secondary generalized motor (SGMS) seizures. Progesterone was well tolerated by 23 of the 25 women and had readily reversible dose-related side effects of asthenia and emotional depression in two. Eighteen women (72%) experienced a decline in seizure frequency during a 3-month treatment period compared with the 3 months prior to therapy (p < 0.01). Average daily CPS frequency declined by 54% (p < 0.01), SGMS by 58% (p < 0.02).


Deep anterior resection with circular stapled anastomosis of congenital megacolon: clinical results.
This report deals with instrumental suture in the deep anterior rectum resection when treating Hirschsprung's disease. The concept includes: ensuring diagnosis, cleaning the intestines, antibiotic prophylaxis, intraoperative testing of the anastomosis and postoperative X-ray by a standardized technique. Our first experience in the use of the stapler was gained with ten patients. One case of anastomosis insufficiency occurred. With the stapler apparatus, safe anastomosis was possible. The stapler facilitates anastomoses in deep resections. Therefore, a deep resection of the pathological segment in Hirschsprung's disease is possible. The practicality of the stapler and the favourable clinical experience justify the continued application of the instrumental suture technique in children's surgery.


The influence of extent and local management on the outcome of radiotherapy for brain metastases.
The results of cranial irradiation for brain metastases in 164 consecutive patients have been reviewed to evaluate a policy of localized high dose irradiation for solitary metastases. Fifty of the 164 patients receiving whole brain irradiation (35 Gy in 15 daily fractions) were selected for boosts delivering 15 Gy in 8 daily fractions to the site of solitary deposits. No difference in overall survival or the incidence of death from progressive brain metastases was seen between the patients receiving a boost and those who did not. Overall median survival was 112 days with 62% of patients dying from metastatic disease outside the brain. Factors associated with increased survival were early response to radiotherapy and, in breast cancer patients, a disease-free interval of 2 years. Palliation of presenting symptoms was achieved in 86% of patients at 3 weeks from starting radiotherapy. It is concluded that whereas whole brain irradiation results in useful symptom control, there is no advantage for high dose treatment in the majority of patients with solitary metastases even in the absence of metastatic disease elsewhere.


Molecular and biochemical analysis of phthalate and terephthalate degradation by Rhodococcus sp. strain DK17.
Alkylbenzene-degrading Rhodococcus sp. strain DK17 is able to utilize phthalate and terephthalate as growth substrates. The genes encoding the transformation of phthalate and terephthalate to protocatechuate are organized as two separate operons, located 6.7kb away from each other. Interestingly, both the phthalate and terephthalate operons are induced in response to terephthalate while expression of the terephthalate genes is undetectable in phthalate-grown cells. In addition to two known plasmids (380-kb pDK1 and 330-kb pDK2), a third megaplasmid (750-kb pDK3) was newly identified in DK17. The phthalate and terephthalate operons are duplicated and are present on both pDK2 and pDK3. RT-PCR experiments, coupled with sequence analysis, suggest that phthalate and terephthalate degradation in DK17 proceeds through oxygenation at carbons 3 and 4 and at carbons 1 and 2 to form 3,4-dihydro-3,4-dihydroxyphthalate and 1,2-dihydro-1,2-dihydroxyterephthalate, respectively. The 3,4-dihydroxyphthalate pathway was further corroborated through colorometric tests. Apparently, the two dihydrodiol metabolites are subsequently dehydrogenated and decarboxylated to form protocatechuate, which is further degraded by a protocatechuate 3,4-dioxygenase as confirmed by a ring-cleavage enzyme assay.


Critical assessment of the spectroscopic activity assay for monitoring trypsin activity in organic-aqueous solvent.
Solvent-assisted protein digestion involves enzymatic hydrolysis in mixed aqueous-organic solvents. With trypsin, acetonitrile is the modifying solvent of choice, recommended at concentrations from 10 to 80% to improve protein sequence coverage in mass spectrometry. Spectroscopic activity assays employing substrate mimics such as N-benzoyl arginine ethyl ester (BAEE) appear to show a relative enhancement of trypsin activity in mixed solvent systems. However, as reported here, the changing solvent polarity induces bias in the absorbance measurement, lending upward of 35% error in the apparent trypsin activity as the acetonitrile is raised to 70%. Furthermore, time-dependent spectroscopic and mass spectrometric measurements reveal a progressive deactivation of trypsin over a 5- to 10-min period in as little as 30% acetonitrile.


Physical and pharmacokinetic characterization of Soluvec™, a novel, solvent-free aqueous ivermectin formulation.
Purpose: To describe application of the Quicksol™ solvent-free approach to solubilize ivermectin (IVM). Methods: Lyophilized IVM complexed with hydroxypropyl-β-cyclodextrin (HP-β-CD) was resolubilized in aqueous polysorbate-80, generating Soluvec™. Lyophilizate was examined by Fourier-transform infrared spectroscopy and differential scanning calorimetry; Soluvec, by dynamic light scattering. Pharmacokinetics was evaluated in dogs allocated to subcutaneous (SC) or intramuscular (IM) Soluvec or oral IVM. Results: IVM in Soluvec was tightly bound by HPβCD, forming nearly monodisperse 28 nm particles with solubility ∼2500-times that of free IVM. SC and IM Soluvec increased IVM exposure, peak IVM and extended duration of IVM exposure, versus oral dosing. Conclusion: The Quicksol method generated Soluvec, a concentrated aqueous parenteral IVM formulation with pharmacokinetic properties suitable for veterinary or human use.


First case series on the use of imiquimod for morphoea.
Morphoea is characterized by fibrosis, which is mediated by cytokines including transforming growth factor (TGF)-beta. Our objective was to use imiquimod 5% cream (Aldara), an inducer of interferon-gamma, known to inhibit TGF-beta, to treat morphoea. Patients with morphoea were treated with imiquimod and evaluated during their follow-up visits to 6 months. The dyspigmentation, induration and erythema of 12 patients with morphoea lesions improved. The histology of the skin also showed a decrease in dermal thickness. This is the first case series describing the successful application of imiquimod in the management of morphoea.


Inclusive Practices toward LGBT Older Adults in Healthcare and Social Services: A Scoping Review of Quantitative and Qualitative Evidence.
Objectives: To determine what constitutes inclusive practices toward LGBT older adults in healthcare and social services and the supportive competencies of these practices. Methods: A comprehensive scoping review of the existing literature was conducted.Results: To be competent when addressing LGBT elder needs, personnel must have knowledge on LGBT realities, openness and be able to put certain skills into practice.Conclusions: Given the limited powers conferred on them by their respective positions, institutions must also play a role in encouraging the inclusion of LGBT older adults. Beyond the importance of taking into account the three dimensions of competence in staff training, changes must be made at all levels of the organizational structures.Clinical implications: Inclusive practices require 1) Educating personnel on the diversity of the life journeys of LGBT older adults; 2) Promoting relational and communication skills and open, friendly attitudes toward the diversity of LGBT identities; 3) Developing policies and procedures to create and maintain inclusive and safe environments for LGBT older adults.


Beta-amyloid protein structure determines the nature of cytokine release from rat microglia.
Activated microglia represent a major source of inflammatory factors in Alzheimer's disease and a possible source of cytotoxic factors. beta-Amyloid (Abeta) peptide, the predominant component in amyloid plaques, has been shown to activate microglia and stimulate their production of inflammatory factors. The present study was performed to analyze the responses of microglia to different forms of Abeta, with regard to release of the proinflammatory cytokines interleukin-1alpha (IL-1alpha), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), IL-6, and interferon-gamma (IFN-gamma), as well as the IL-1 receptor antagonist (IL-1ra). Primary cultures of microglia from rat neonatal cerebral cortex were incubated with freshly dissolved Abeta1-40 or Abeta1-42, Abeta1-40 fibrils, Abeta1-40 betaamy balls, or vehicle. Abeta1-40 fibrils did not significantly stimulate any of these cytokines. Freshly dissolved Abeta1-40 resulted in a marked increase in the release of IL-1beta, and freshly dissolved Abeta1-42 significantly stimulated both IL-1alpha and IFN-gamma secretion. The Abeta1-40 betaamy balls stimulated the secretion of IL-1alpha and IL-1beta. Incubation with Abeta peptides did not affect the secretion of IL-1ra, IL-6, or TNF-alpha. In the case of IL-1beta, the response is correlated with the presence of Abeta peptide as monomers and oligomers.


Glossy mutants of maize. VIII. Accumulation of fatty aldehydes in surface waxes of gl5 maize seedlings.
In corn seedlings (Zea mays L.) homozygous for the mutation gl5, the surface waxes are characteristically altered. In this mutant the main wax constituents (83.5%) are aldehydes while in the normal waxes alcohols predominate (62.7%). Moreover, in the normal waxes aldehydes and alcohols are made up mainly of the C32 term (99%), whereas in gl5 waxes the principal aldehyde is still C32 (90.7%) but the free alcohol composition pattern is noticeably modified. Here the predominant terms are C24, C26, and C28, with C32 representing only 16.6% of the total. The results indicate that the mutant induces a block in the synthesis of fatty alcohols while accumulating fatty aldehydes, the substrates from which the alcohols originate.


Expression of NTPDase1 and caveolins in human cardiovascular disease.
Pathological circumstances like inflammation or ischemic insult facilitate the release of adenine nucleotides from several types of cells. These extracellular nucleotides are rapidly converted to adenosine by ectonucleotidases, mainly ectonucleoside triphosphate diphosphohydrolase1 (NTPDase1/CD39) and CD73. NTPDase1/CD39 can interact with caveolins, structural proteins of signal-transducing microdomains termed caveolae. Caveolins are thought to have physiological roles in heart ageing and cardiac diseases. The aim of this study was to investigate the expression of NTPDase1 together with caveolins in chronic human cardiovascular diseases and elucidate their role in human heart. The HPLC analysis showed significant increase in ATPase activity in pathological samples from patients with ischemic heart disease. Immunostaining also showed alterations in the expression and distribution of NTPDase1. Caveolin-1 and caveolin-2 expression was much alike in control and pathological cases, while expression of caveolin-3 was lower in pathological samples. Changes in the expression of NTPDase1 and caveolins seem to be independent of human cardiovascular disease.


Higher vitamin B6 status is associated with improved survival among patients with stage I-III colorectal cancer.
Folate-mediated 1-carbon metabolism requires several nutrients, including vitamin B6. Circulating biomarker concentrations indicating high vitamin B6 status are associated with a reduced risk of colorectal cancer (CRC). However, little is known about the effect of B6 status in relation to clinical outcomes in CRC patients. We investigated survival outcomes in relation to vitamin B6 status in prospectively followed CRC patients. A total of 2031 patients with stage I-III CRC participated in 6 prospective patient cohorts in the international FOCUS (folate-dependent 1-carbon metabolism in colorectal cancer recurrence and survival) Consortium. Preoperative blood samples were used to measure vitamin B6 status by the direct marker pyridoxal 5'-phosphate (PLP), as well as the functional marker HK-ratio (HKr)[3'-hydroxykynurenine: (kynurenic acid + xanthurenic acid + 3'-hydroxy anthranilic acid + anthranilic acid)]. Using Cox proportional hazards regression, we examined associations of vitamin B6 status with overall survival (OS), disease-free survival (DFS), and risk of recurrence, adjusted for patient age, sex, circulating creatinine concentrations, tumor site, stage, and cohort. After a median follow-up of 3.2 y for OS, higher preoperative vitamin B6 status as assessed by PLP and the functional marker HKr was associated with 16-32% higher all-cause and disease-free survival, although there was no significant association with disease recurrence (doubling in PLP concentration: HROS, 0.68; 95% CI: 0.59, 0.79; HRDFS, 0.84; 95% CI: 0.75, 0.94; HRRecurrence, 0.96; 95% CI: 0.84, 1.09; HKr: HROS, 2.04; 95% CI: 1.67, 2.49; HRDFS, 1.56; 95% CI: 1.31, 1.85; HRRecurrence, 1.21; 95% CI: 0.96,1. 52). The association of PLP with improved OS was consistent across colorectal tumor site (right-sided colon: HROS, 0.75; 95% CI: 0.59, 0.96; left-sided colon: HROS, 0.71; 95% CI: 0.55, 0.92; rectosigmoid junction and rectum: HROS, 0.61; 95% CI: 0.47, 0.78). Higher preoperative vitamin B6 status is associated with improved OS among stage I-III CRC patients.


Assessing the size-dependent effects of microplastics on zebrafish larvae through fish lateral line system and gut damage.
This study evaluated the size-dependent effects of high-density polyethylene (HDPE) fragments in zebrafish. Larvae were exposed to HDPE microplastic (MP) in three sizes, small (14.12 μm), medium (80.32 μm), and large (120.97 μm), at 20 mg/L. Size-dependent effects in terms of MP intake, subsequent gut damage, and behavioral changes were observed. The results showed that HDPE exposure did not affect the survivability of zebrafish larvae but caused two significant changes. First, exposure to large MPs caused the most serious damage to hair cells and mechanosensory receptors in the fish's lateral line system. Second, exposure to MPs < 100 μm resulted in their ingestion by larvae, thereby causing morphological changes in the gastrointestinal tract. All larvae exposed to MPs showed behavioral pattern changes associated with size differences. This study improves our understanding of the effects of MPs on aquatic organisms and highlights the need to implement efficient strategies for plastic waste management.


Peripheral clonal deletion of superantigen-reactive T cells is enhanced by cortisone.
The T cell receptor (TcR) V beta-specific expansion, deletion and induction of nonresponsiveness among murine T cells responding to superantigens in the periphery has been well characterized. Here we demonstrate that clonal deletion of staphylococcal enterotoxin (SE) B-reactive V beta 8.2+ cells can be significantly increased when mice are injected with hydrocortisone (HC) following superantigen stimulation in vivo. The induced sensitivity to HC persists for at least 30 days after SEB injection, making it unlikely that proliferating cells were uniquely responsible for the enhanced deletion. Superantigen-induced HC sensitivity was a general phenomenon and could also be observed among V beta 11+ cells after the injection of SEA. Experiments conducted on thymectomized mice indicated that HC-sensitive, SEB-responsive cells could not be accounted for by rapidly produced, immature lymphocytes recently exported from the thymus. Further, V beta 8.1+ peripheral lymphocytes from TcR transgenic mice expressing the Mls-1a superantigen were sensitive to HC. These results imply that the majority of cells remaining after superantigen-induced clonal expansion and deletion in vivo have indeed reacted with the superantigen. Implications for differential superantigen recognition by T cells expressing the same TcR V beta domain, perhaps due to a significant V alpha contribution to the interaction in vivo, are discussed.


Serological survey on some pathogens in wild brown hares (Lepus europaeus) in Central Italy.
To determine the exposure of wild brown hares [Lepus europaeus (L. europaeus), pallas] to Anaplasma phagocytophilum (A. phagocytophilum), Borrelia burgdorferi (B. burgdorferi) sensu lato, Encephalitozoon cuniculi (E. cuniculi), Leishmania sp., Neospora caninum (N. caninum) and Toxoplasma gondii (T. gondii). Two hundred twenty-two blood serum samples of wild brown hares captured in protected areas of the province of Pisa (Central Italy) were tested to detect antibodies against the reported pathogens. Thirty one (14.0%) animals resulted positive for at least one tested agent, with antibody titres ranging from 1:20 to 1:320. In particular, 13 (5.8%) samples were positive to B. burgdorferi s.l., 11 (4.9%) to N. caninum, 3 (1.3%) to T. gondii, 2 (0.9%) to A. phagocytophilum and 2 (0.9%) to Leishmania sp. No samples scored positive to E. cuniculi. Four animals (14.8%) resulted coinfected with 2 different pathogens. The obtained results showed that B. burgdorferi s.l. N. caninum, T. gondii, A. phagocytophilum and Leishmania sp. circulate in wild brown hares in Central Italy, suggesting a possible role of L. europaeus as reservoir of these pathogens. The obtained results showed that autochthonous wild brown hares living in Central Italy have been exposed to several pathogens circulating in this area, suggesting a possible role of L. europaeus as reservoir.


The effect of RU 41740 on the in vitro maturation of human B-cells.
We have tested the effect of a Klebsiella pneumoniae extract, RU 41740, and its lipopolysaccharidic fraction (LPS-LAP) on the in vitro activation of human B-cells. Two models have been used: the polyclonal activation induced by pokeweed mitogen and the specific antibody production to mannan, a polysaccharide extracted from the cell wall of Candida albicans. We have shown that RU 41740, and especially its lipopolysaccharidic fraction, significantly increases the production of immunoglobulins and specific antibodies. This effect is mediated by T-lymphocytes, since preincubation of isolated T-cells, but not of non-T-cells, resulted in the same effect. Together with the known enhancing effect of LPS-LAP on T-cell proliferation, these data strongly suggest that LPS-LAP increases the production of B-cell-activating lymphokines by T-cells.


Defibrotide decreases cholesterol amount in hypercholesterolemic rabbit aorta, with no modification of plasma or lipoprotein cholesterol.
Defibrotide, (D) an antithrombotic agent, when administered i.v. to cholesterol-fed rabbits decreased cholesterol in the aorta without changing total plasma cholesterol, triglyceride or phospholipid, nor the cholesterol, triglyceride, phospholipid and protein of plasma lipoproteins. Platelet aggregation was decreased in rabbits treated with D. There were fewer vascular lesions in the hearts and kidneys of animals treated with D than in animals fed cholesterol and treated with placebo. These data suggest that the antithrombotic activity of D and its ability to reduce platelet sensitivity could help to reduce the amount of cholesterol in the cardiovascular system in atherosclerosis-prone situations.


Drinking saccharin increases food intake and preference--II. Hydrational factors.
Rats that drink saccharin solution increase their short-term food intake and develop a preference for flavored food eaten when saccharin is ingested. In this paper, we examined whether these changes in feeding behavior were due to overhydration resulting from drinking hyposmotic saccharin solution. Consistent with this possibility, the short-term food intake of rats was increased by drinking hyposmotic 0.2% saccharin dissolved in water, unaffected by drinking isosmotic 0.9% NaCl, and decreased by drinking 0.2% saccharin dissolved in 0.9% NaCl. In addition, rats showed a sustained increase in saccharin-induced food intake after antidiuretic hormone treatment, which was designed to exacerbate their positive water balance. Less consistent with a hydrational explanation of saccharin-induced feeding was the finding that rats drinking only 2ml 0.2% saccharin solution increased food intake. Also, gastric intubation of similar volumes of water produced a small, transient increase in feeding behavior, which was apparent after the first intubation only and could not be preserved by adding water-contingent flavors to the food. Taken together, these results suggest that the hydrational effects of drinking hyposmotic saccharin solution contribute to, but cannot account for, the increase in food intake. Hydration had no observable influence on the acquisition of flavored food preference.


Prolonged STAT1 activation in neurons drives a pathological transcriptional response.
Neurons require physiological IFN-γ signaling to maintain central nervous system (CNS) homeostasis, however, pathological IFN-γ signaling can cause CNS pathologies. The downstream signaling mechanisms that cause these drastically different outcomes in neurons has not been well studied. We hypothesized that different levels of IFN-γ signaling in neurons results in differential activation of its downstream transcription factor, signal transducer and activator of transduction 1 (STAT1), causing varying outcomes. Using primary cortical neurons, we showed that physiological IFN-γ elicited brief and transient STAT1 activation, whereas pathological IFN-γ induced prolonged STAT1 activation, which primed the pathway to be more responsive to a subsequent IFN-γ challenge. This is an IFN-γ specific response, as other IFNs and cytokines did not elicit such STAT1 activation nor priming in neurons. Additionally, we did not see the same effect in microglia or astrocytes, suggesting this non-canonical IFN-γ/STAT1 signaling is unique to neurons. Prolonged STAT1 activation was facilitated by continuous janus kinase (JAK) activity, even in the absence of IFN-γ. Finally, although IFN-γ initially induced a canonical IFN-γ transcriptional response in neurons, pathological levels of IFN-γ caused long-term changes in synaptic pathway transcripts. Overall, these findings suggest that IFN-γ signaling occurs via non-canonical mechanisms in neurons, and differential STAT1 activation may explain how neurons have both homeostatic and pathological responses to IFN-γ signaling.


Structural features of the nicotinic acetylcholine receptor revealed by antibodies to synthetic peptides.
Antibodies were raised to the amino- and carboxy-terminal decapeptides of Torpedo californica acetylcholine receptor. Structural studies of the native receptor using the antipeptide antibodies as probes proved the existence of the carboxy terminal sequence in the alpha subunit predicted from its cDNA sequence and supported structural models of the native receptor that place the carboxy termini on the intracellular side. The amino termini of the subunits were not accessible on the surface of native receptor.


Differences in Fungal Disease Dynamics in Co-occurring Terrestrial and Aquatic Amphibians.
The fungal pathogen, Batrachochytrium dendrobatidis (Bd), has devastated biodiversity and ecosystem health and is implicated as a driver of mass amphibian extinctions. This 100-year study investigates which environmental factors contribute to Bd prevalence in a fully terrestrial species, and determines whether infection patterns differ between a fully terrestrial amphibian and more aquatic host species. We performed a historical survey to quantify Bd prevalence in 1127 Batrachoseps gregarius museum specimens collected from 1920 to 2000, and recent data from 16 contemporary (live-caught) B. gregarius populations from the southwestern slopes of the Sierra Nevada mountains in California, USA. We compared these results to Bd detection rates in 1395 historical and 1033 contemporary specimens from 10 species of anurans and 427 historical Taricha salamander specimens collected throughout the Sierra Nevada mountains. Our results indicate that Bd dynamics in the entirely terrestrial species, B. gregarius, differ from aquatic species in the same region in terms of both seasonal patterns of Bd abundance and in the possible timing of Bd epizootics.


Changes in brain dopamine receptors in MPTP parkinsonian monkeys following L-dopa treatment.
Twenty-two monkeys (Macaca fascicularis) were utilized in this study. Ten animals were rendered parkinsonian with serial injections of MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine). Five of these parkinsonian monkeys received L-dopa/carbidopa treatment, and five animals did not. The remaining twelve animals did not receive MPTP. Eight of these animals received no L-dopa treatment, two animals were treated chronically with L-dopa/carbidopa and two animals received L-dopa/carbidopa only on the day of sacrifice. All animals were given weekly scored neurologic examinations throughout the study. Their movement was quantitated in an activity box. All animals were sacrificed by an overdose of sodium pentobarbital. The parkinsonian animals were sacrificed 107-355 days after their last MPTP injection. The brains were removed and frozen. Punch samples were taken from the caudate and putamen for tissue dopamine determination. Selected areas of the basal ganglia were cut into 20 microns sections for quantitative receptor autoradiography. The density of D1 and D2 receptors was evaluated in the basal ganglia of these animals at the level of the anterior commissure. For the D2 assay, total binding was determined using various concentrations of [3H]spiperone in buffer containing 300 nm mianserin. For the D1 assay, total binding was determined using various concentrations of [3H]SCH-23390. Tissue isotope concentration was determined from the autoradiographs. The MPTP parkinsonian monkeys showed a mean striatal dopamine depletion of 93.5% and a mean clinical score of 9.0. The untreated parkinsonian monkeys demonstrated an increase in the number of D2 sites as compared to controls. This increase was greatest in the lateral putamen.(ABSTRACT TRUNCATED AT 250 WORDS)


Social disorganization/social fragmentation and risk of depression among older people in Japan: multilevel investigation of indices of social distance.
Previous studies reported that social disorganization/fragmentation could predict mental well-being of residents in a community. The aim of this study is to examine how area and individual level of social distance could predict likelihood of mental health among older people in Japan. We empirically derived an index of "social distance" by taking averaged differences in sociodemographic characteristics that are income, education, hometown of origin, the duration of residency, and life stage, between the study participants and their neighbors. We used the study participants (n = 9147) from the Aichi Gerontological Evaluation Study, which targeted residents with aged 65 years or over in a central part in Japan. Depressive symptoms of the study participants were assessed using the short version of the Geriatric Depression Scale (GDS-15). We also tested if area-level social capital would moderate the association between social distance and depressive symptoms. Using multilevel analyses, we found that higher social distance from neighbors was associated with increased depressive symptoms, independently of respondents' own values of income and educational attainment. At the individual level, each standard deviation in income-based and education-based social distance was associated with an odds ratio for depressive symptoms of 1.15 (95% CI: 1.01-1.30) and 1.17 (95% CI: 1.03-1.32), respectively. However, the area-aggregated indices of social distance were not associated with depressive symptoms. Additionally, area-level social capital indicating higher levels of trust between neighbors and social participation, buffered the adverse effect of social distance on depressive risk. In an instance of the "dark side" of social capital, we also found that stronger social cohesion increased depressive symptoms for residents whose hometown of origin differed from the communities where they currently resided.


Moonlighting vacuolar protease: multiple jobs for a busy protein.
In this Genomics Era with a wealth of annotated sequence data, it is easy to pigeonhole a protein into a particular function. However, Noa Matarasso et al. recently found a vacuolar protease that can also function as a transcription factor. This work illustrates that a protein can serve multiple roles in a cell, raising intriguing questions as to the extent that genomic information can be deciphered de novo.


Optimized inhalation aerosols. II. Inertial testing methods for particle size analysis of pressurized inhalers.
Pressurized metered dose inhaler (MDI) output from three different albuterol formulations was characterized using three inertial separation devices. Results were compared for the Delron six-stage cascade impactor (DC16), the Andersen Mark II eight-stage impactor (AC18), and Copley's twin-stage liquid impinger (LI). None of the devices tested in this study was ideal in all respects. All devices could differentiate between formulations in terms of respirable doses (albuterol amount with aerodynamic diameters less than 5.5 through 6.4 microns). Only the high-flow rate LI could differentiate among all three formulations when data were presented in terms of respirable percentage (RP) of drug collected. Values for RP were in excellent agreement for the independently calibrated impactors when the same evaporation chamber was used atop the impactors. The LI appeared to overestimate values for RP in vivo. Results are discussed in light of the debate surrounding the revision of USP aerosol testing requirements. Rigorous specifications for evaporation chambers and methodologies are necessary for meaningful inter- and intra-laboratory comparison of results when any of these devices are used.


Neural trans-differentiation potential of hepatic oval cells in the neonatal mouse brain.
Although the existence of the hepatic oval cell (HOC), the liver stem cell has been known for almost 70 years, little is known about the potential for this adult stem cell to trans-differentiate into cells of other tissues. While their origin remains enigmatic, HOCs share many similarities with hematopoietic stem cells. Recent studies have revealed that a small percentage of HOCs can arise from a bone marrow-derived stem cell source. Here we report that, like bone marrow stem cells, HOCs can survive transplantation to the neonatal mouse brain and show signs of trans-differentiation by adopting the morphology and antigenic phenotype of both macro- and microglia cells. Trans-differentiated microglia cells were functional, showing active phagocytosis when cotransplanted with latex microbeads in vivo. In addition to glial markers, a small number of transplanted HOCs were immunopositive for neuronal markers, but displayed ambiguous phenotype, making their characterization difficult.


How droplets move on laser-structured surfaces: Determination of droplet adhesion forces on nano- and microstructured surfaces.
Lateral adhesion forces are a fundamental property of liquid-solid interactions and a key aspect of dynamic droplet mobility. But, commonly applied conventional wetting analysis is limited to static and quasi-static methods and cannot resolve dynamic and spatial liquid-solid interactions. However, droplet mobility is assumed to be affected by chemical and topographic surface inhomogeneities introduced by femtosecond laser treatment. In this study, we used a customized droplet adhesion force instrument to determine lateral adhesion forces on various femtosecond laser-structured surface designs to obtain a deeper understanding of the dynamic droplet motion with regard to chemical and topographic surface features. We show that the droplet motion was highly affected by the chemical and topographical surface design and local inhomogeneities. The droplet mobility on femtosecond laser-structured surfaces could be classified into a static, a transfer, and a kinetic regime, which is essential for designing surfaces with extreme wetting characteristics and a wide range of scientific and industrial processes. Furthermore, with proper tailoring of surface structures and chemical modification, we were able to provoke adhesion forces on self-organized laser microstructures similar to those found on the natural lotus leaves.


The relationship between brachytherapy dose and outcome in patients with bulky endocervical tumors treated with radiation alone.
To evaluate the relationship between brachytherapy dose and outcome in patients treated with external radiotherapy (40 Gy to the whole pelvis) and intracavitary radium therapy for bulky endocervical tumors. Between 1962 and 1985, 98 patients with Stage IB-IIB bulky endocervical carcinomas (> or = 6 cm in diameter) treated with radiotherapy alone received 40 Gy to the whole pelvis followed by 2 or more intracavitary treatments. Twenty-five patients received < 6000 mg-hr of intracavitary treatment and 73 received > or = 6000 mg-hr (an average dose to point A of approximately 49 Gy). Brachytherapy exposures ranged from 4800-7885 mg-hrs. Patients who received < 6000 mg-hr tended to have unfavorable (narrow) vaginal anatomy (p < 0.01) and to be treated in the later years of the study (p < 0.01). The high-dose group included a somewhat greater proportion of patients with positive lymphangiograms or poor responses to initial external beam treatment. Despite having somewhat more favorable tumors, patients who received less than 6000 mg-hr had a higher rate of pelvic disease recurrence at 5 years (33%) than those who received higher doses (16%) (p = 0.03). Actuarial survival rates at 5 years were 44% and 60% for the low- and high-dose groups, respectively (p = 0.14). Among those who received more than 6000 mg-hr, there was no significant relationship between brachytherapy dose and pelvic disease control. Calculated actuarially, the rate of major (> or = grade 3) complications at 5 years was 23% in the low-dose group and 10% in the high-dose group (p = 0.1). The relatively high incidence of pelvic disease recurrence and complications in patients who receive less than 6000 mg-hr reflects the narrow therapeutic window for complication-free pelvic disease control in patients with bulky central disease and unfavorable normal tissue anatomy. The results also demonstrate a high pelvic control rate and acceptable morbidity in patients with favorable anatomy treated with high-dose radiotherapy alone.


In vitro effects of cholesteryl butyrate solid lipid nanospheres as a butyric acid pro-drug on melanoma cells: evaluation of antiproliferative activity and apoptosis induction.
Literature data show that butyric acid derivatives bear a dose-dependent differentiative anti-proliferative activity on cancer cell lines and that apoptosis induction may play a major role. Although it was recently shown that solid lipid nanospheres (SLNs) are a suitable tool for several in vivo drug administration routes, there is little available information on melanoma cell lines. This study was aimed at evaluating the anti-proliferative and apoptotic in vitro effects of cholesteryl butyrate (chol-but) SLNs on melanoma cells. Increasing concentrations of chol-but SLNs were used to test two melanoma cell lines. Both cell lines were treated with Na-butyrate (Na-but) and chol-but SLNs for viability. Those tested with chol-but SLNs were more effective than Na-butirate (3 to 72 h). The apoptotic effects of chol-but SLNs were evaluated between 3 and 72 h by annexin-V (ANX-V)/propidium iodide (PI) staining and the antiproliferative effect by PI staining. Apoptosis anti-proliferative-regulatory proteins as bcl-2, Fas/APO1 (CD95) and PCNA (PC10) were also investigated. Flow cytometric analyses evidenced a G(0/1)-S transition block and a 'sub-G(0/1)' apoptotic peak from 0.5 to 1.0 mM butyric acid. In ANX-V/PI flow cytometric staining, a dose- and time-dependent increase in the apoptotic cell percentage (ANX-V+) coupled with a down-regulation of PC10 and bcl-2 and a parallel up-regulation of Fas/APO1 (CD95) were found in both lines started after 3 to 24 h of chol-but SLNs treatment. Results show that chol-but SLNs exerts a dose/time-dependent effect in melanoma cell apoptosis induction between 3 and 24 h and a dose but not time-dependent effect after 24 h of treatment.


Total white blood cell counts and LPS-induced TNF alpha production by monocytes of pregnant, pseudopregnant and cyclic rats.
Pregnancy in the rat may be associated with an activated innate immune system. Therefore, we investigated monocyte function as well as total white blood cell (WBC) counts during the follicular phase of the ovarian cycle, pregnancy and pseudopregnancy in the rat. Rats were equipped with a permanent jugular vein cannula, and 0.43 ml blood samples were taken from this cannula during the 4 days of the regular oestrus cycle of the rat (n=12). Thereafter, six rats were rendered pregnant, and the other six rats were rendered pseudopregnant according to standard methods. Blood samples were withdrawn from the cannula on days 4, 7 and 11 of pseudopregnancy and on days 4, 7, 11 and 20 of pregnancy. From each blood sample, 0.4 ml was stimulated with lipopolysaccharide (LPS) and monocyte intracellular cytokine production measured using flow cytometry. 30 microl of the blood was used to measure WBC counts and differential WBC counts. The results showed that the number of WBC was significantly increased only on day 11 of pregnancy compared with the follicular phase, and that this was due to the increased numbers of polymorphonuclear (PMN) cells. The percentage of TNF alpha-producing monocytes was increased on all days of pseudopregnancy and on day 11 of pregnancy. The fact that the percentage of monocytes producing TNF alpha upon an LPS stimulus was increased during the post-implantation phase of pregnancy and during pseudopregnancy as compared to the follicular phase may indicate that these conditions are proinflammatory conditions. For the post-implantation phase of pregnancy, this is once more stressed by the increased numbers of WBC and PMN.


Impaired CD95-mediated apoptosis in autoimmunity and occurrence of a p22 Caspase-8 cleavage product in JIA.
Altered apoptosis can lead to autoimmune diseases such as systemic lupus erythematosus (SLE). Juvenile idiopathic arthritis (JIA) is another autoimmune disease characterized by autoinflammation. During this process activated T-cells accumulate in the synovial fluid. We hypothesized that resistance to CD95-mediated apoptosis could contribute to autoimmune phenotypes. We isolated highly activated T cells (CD45RO+ and CD95+) by magnetic separation from healthy controls, JIA patients and patients with other autoimmune diseases. In these purified cells, apoptosis was induced by stimulation with recombinant human soluble CD95 ligand (rhsCD95L) or dexamethasone and analyzed by flow cytometry. In addition, cleavage and expression of apoptosis mediators (Caspase-8 and -3) and regulators (FLIP, Bcl-2, Bcl-xL) were analyzed in mononuclear cells using immunoblot technique. Apoptosis upon CD95 stimulation, but not dexamethasone treatment, was reduced in JIA patients and patients with other autoimmune diseases compared to healthy controls. Additionally we observed a non-canonical cleavage pattern of Caspase-8 resulting in a p22 fragment and high expression of FLIP in SFMCs of patients with JIA. Expression and cleavage of proteins of the CD95 pathway is altered in JIA providing a possible explanation for resistance against death receptor-mediated apoptosis. Dexamethasone-induced apoptosis, however, is intact arguing against a general defect in apoptosis. The implications of the p22 fragment regarding apoptosis have to be further analyzed. The strong expression of FLIPShort in SFMCs may as well result from the highly activated status of the cells or be a feature of autoimmunity.


Folate deficiency presenting as pyrexia: a case report.
Folate deficiency is an uncommon cause of pyrexia. We describe the case of a 29-year-old male who presented with a pyrexial illness subsequently attributed to megaloblastic anaemia secondary to severe folate deficiency, after exclusion of other infective or inflammatory causes. A temperature chart documenting the course of the patient's pyrexia is presented and potential pathophysiological mechanisms are proposed. Folate deficiency is a reversible cause of pyrexia that should be considered in any patient who presents with a pyrexial illness of unknown cause.


Congenital disorders of glycosylation.
Congenital disorders of glycosylation (CDG) are genetic diseases due to defects in the synthesis or the attachment of the glycan moiety of glycoproteins and glycolipids. They can be divided into four groups: disorders of protein N-glycosylation, disorders of protein O-glycosylation, disorders of lipid glycosylation, and disorders of other glycosylation pathways and of multiple glycosylation pathways. Of the more than 40 reported CDG, some 80% are neurological or have an important neurological component. By far the most common neurological CDG is phosphomannomutase 2 deficiency. Isoelectrofocusing of serum transferrin, the most widely used screening test, picks up only CDG associated with sialic acid deficiency of N-linked glycans. Predominant neurological signs and symptoms are psychomotor retardation, epilepsy, hypotonia, hyporeflexia, strabismus, retinitis pigmentosa, polyneuropathy, myopathy, and cerebellar hypotrophy/hypoplasia. All known neurological CDG have an autosomal recessive inheritance except for IAP-CDG, an X-linked pure mental retardation syndrome. No curative or effective treatment is available for neurological CDG. Since at least 1% of the genome is involved in glycosylation, it is likely that the large majority of CDG is yet to be discovered. In 2008, a novel nomenclature was introduced using the gene symbol followed by -CDG, e.g., CDG-Ia becomes PMM2-CDG. CDG should be looked for in any unexplained neurological syndrome.


Cardiac and skeletal muscle disorders caused by mutations in the intracellular Ca2+ release channels.
Here we review the current knowledge about the mutations of the gene encoding the cardiac ryanodine receptor (RyR2) that cause cardiac arrhythmias. Similarities between the mutations identified in the RyR2 gene and those found in the gene RyR1 that cause malignant hyperthermia and central core disease are discussed. In vitro functional characterization of RyR1 and RyR2 mutants is reviewed, with a focus on the contribution that in vitro expression studies have made to our understanding of related human diseases.


Bhlha9 regulates apical ectodermal ridge formation during limb development.
Split hand/foot malformation (SHFM) and SHFM combined with long-bone deficiency (SHFLD) are congenital dysgeneses of the limb. Although six different loci/mutations (SHFM1-SHFM6) have been found from studies on families with SHFM, the causes and associated pathogenic mechanisms for a large number of patients remain unidentified. On the basis of the identification of a duplicated gene region involving BHLHA9 in some affected families, BHLHA9 was identified as a novel SHFM/SHFLD-related gene. Although Bhlha9 is predicted to participate in limb development as a transcription factor, its precise function is unclear. Therefore, to study its physiological function, we generated a Bhlha9-knockout mouse and investigated gene expression and the associated phenotype in the limb bud. Bhlha9-knockout mice showed syndactyly and poliosis in the limb. Moreover, some apical ectodermal ridge (AER) formation related genes, including Trp63, exhibited an aberrant expression pattern in the limb bud of Bhlha9-knockout mice; TP63 (Trp63) was regulated by Bhlha9 on the basis of in vitro analysis. These observations suggest that Bhlha9 regulates AER formation during limb/finger development by regulating the expression of some AER-formation-related genes and abnormal expression of Bhlha9 leads to SHFM and SHFLD via dysregulation of AER formation and associated gene expression.


14,15-EET induces the infiltration and tumor-promoting function of neutrophils to trigger the growth of minimal dormant metastases.
Infiltrating neutrophils are known to promote in the development of tumor. However, it is unclear whether and how neutrophils are involved in triggering the growth of dormant metastases. Here we show that 14,15-epoxyeicosatrienoic acid (14,15-EET) can trigger the growth of dormant micrometastases by inducing neutrophilic infiltration and converting neutrophil function. 14,15-EET triggered neutrophil infiltration in metastatic lesions by activating STAT3 and JNK pathways to induce the expression of human IL-8 and murine CXCL15 in corresponding tumor cells. The continuous expression of hIL-8/mCXCL15 was maintained by the sustained and enhanced activation of JNK pathway. 14,15-EET up-regulated miR-155 expression by activating STAT3 and JNK pathways. miR-155 in turn down-regulated the expression of SHIP1 and DET1, thus augmenting the activation of JNK and c-Jun. Moreover, the function of neutrophils was converted from tumor-suppressing to tumor-promoting by 14,15-EET in vivo. By inducing the production of G-CSF/IL-6 in vivo, 14,15-EET induced the enhancement of STAT3 activation in neutrophils to increase MMP-9 expression and decrease TRAIL expression. Neutrophil-derived MMP-9 was required for 14,15-EET to induce angiogenesis during the growth of dormant micrometastases. Depleting neutrophils or inhibiting hIL-8/mCXCL15 up-regulation resulted in the failure of 14,15-EET to promote the development of micrometastases. These findings reveal a mechanism through which the infiltration and tumor-promoting function of neutrophils could be induced to trigger the growth of dormant metastases, which might be a driving force for the tumor recurrence based on dormant metastases.


[The concept of frailty--from phenomenology to therapeutic approaches].
Frailty--the weakness of many (oldest) old persons--induces a lot of human sorrow with functional disabilities as well as health-economic costs. Sarcopenia (loss of muscle mass) is also a form of frailty and a co-factor of two of the "geriatric Is"--immobility and instability. Pathophysiologically, both phenomena show signs of an inflammatory state. Physical activity and a healthy diet can partially reverse these health problems. Therefore, therapeutic strategies emerge to treat these frequent and clinically relevant phenomena.


Increasing hip fracture incidence in California Hispanics, 1983 to 2000.
Hip fracture incidence in non-Hispanic whites (NHW) has decreased nationwide for the past 20 years. Little is known regarding hip fracture incidence among Hispanics, the largest, fastest growing minority in the United States. To assess the change in standardized hip fracture incidence from 1983 through 2000 in California Hispanics relative to other racial groups. Hospitalizations for individuals older than 55 years with hip fracture requiring repair in acute care hospitals. Annual population estimates based on US Census Bureau estimates. Incidence standardized to national gender-age strata. Change in annual incidence calculated by weighted linear regression with robust variance estimates. 372,078 hip fractures were identified. Age-adjusted annual incidence of hip fractures declined by 0.74% per year among women (655 to 568 per 100,000), but was unchanged among men (247 to 238 per 100,000). Among NHW women, the standardized annual incidence fell by 0.6% (4.0 fractures per 100,000) per year. Annual incidence among Hispanic women increased 4.9% (11.1 fractures per 100,000) per year. Annual incidence among Hispanic men increased by 4.2% (4.5 fractures per 100,000) per year and among NHW men by 0.5% (1.2 fractures per 100,000) per year. No significant change occurred among black or Asian women or men. Among California women, hip fracture incidence has doubled among Hispanics since 1983, while remaining unchanged or declining in other groups. Greater attention should be given to identification of individuals at risk for hip fracture and initiation of preventive measures in Hispanic populations.


Neighborhood environment, self-efficacy, and physical activity in urban adolescents.
To test the pathways between perceptions of built environment factors and physical activity in urban youth. Three hundred fifty high school students' perceptions of neighborhood, and barrier self efficacy were measured by a Web survey. Physical activities were assessed using a one-week diary and accelerometers. Land-use mix/accessibility and neighborhood satisfaction had direct pathways to walking. Barrier self-efficacy had a direct pathway to walking. In addition, land use, specifically neighborhood accessibility, influenced adolescents' walking behavior via self-efficacy. Similar pathways were found in MVPA models. Neighborhood factors appear to work together with self-efficacy to facilitate physical activity.


The polymorphisms of extracellular matrix-remodeling genes are associated with pelvic organ prolapse.
Extracellular matrix (ECM) synthesis and metabolism abnormalities may influence the pelvic supporting system and lead to the occurrence and development of pelvic organ prolapse (POP). Genetic polymorphisms of such related genes have been increasingly studied. This study aims to explore the association between the single-nucleotide polymorphisms (SNPs) of genes encoding ECM processing enzymes (a disintegrin and metalloproteinase with thrombospondin motifs [ADAMTSs]), ECM degrading enzymes (matrix metalloproteinases [MMPs]) and their tissue inhibitors of metalloproteinase (TIMPs), and POP. We conducted an association study including 48 women with POP at stages III and IV and 48 women without prolapse in Chinese groups. SNPs were identified using the target region sequencing technique. We performed Fisher's exact tests to assess the association between SNPs and POP in the unadjusted model and logistic regression analysis in the adjusted model, adjusting for delivery and pregnancy. There was a significant association between TIMP2 SNP rs2277698 (odds ratio [OR], 0.37; 95% confidence interval [CI], 0.16-0.82; P = 0.015), ADAMTS13 SNP rs149586801 (OR, 0.18; 95% CI, 0.05-0.69; P = 0.012), and ADAMTS1 SNPs rs370850 and rs422803 (OR, 3.71; 95% CI, 1.35-10.15; P = 0.011 for both), rs402007, rs428785, rs434857, and rs445784 (OR, 2.18; 95% CI, 1.05-4.56; P = 0.038 for the four), and POP in the adjusted model. TIMP2, ADAMTS13, and ADAMTS1 might be candidate genes for POP. Our results provide preliminarily new evidence for future investigation of these genes in the pathophysiology of POP.


First Report of Alternaria tenuissimain Causing Leaf Spot of Celery (Apium graveolens) in China.
Celery (Apium graveolens) is one of the most widely grown vegetables in the world. A survey in Anding District of Gansu Province in 2019 showed that the incidence of celery leaf spot was 25%-45%. The disease mainly occurs in late June and July. The leaf spot is conducive to the onset at high temperature and humidity environment. The initial symptoms were many small light brown, irregular-shaped on the leaves. The lesions gradually enlarged in the later stage of the disease, and multiple lesions coalesced to form large irregular brown spots, eventually the whole leaves died. A 3~4mm leaf tissue was cut from the junction of the diseased leaf and the healthy area, the leaf tisse was surface-sterilized in 1.5% NaClO for 1 min and washed with sterile water. Then, it was incubated on potato dextrose agar (PDA) and obtained the pure culture (Q1). After 5 days of cultivation at 25°C, the fungal colonies were olivaceous to dark olive with white margins and abundant aerial mycelia. The conidia were obclavate or ellipsoid, pale brown, with 3~4 longitudinal septa and 2~7 transverse septa, and measured 20.0 to 50.0 × 3.5 to 14.0μm (n=50). Conidiophores were septate, arising singly, and measured 3.5 to 40.0 × 2.5 to 4.5 μm (n=50). Based on morphological characteristics, the fungus was preliminarily identified as A.tenuissima (Simmons 2007). To further confirm the identification, the internal transcribed spacer region (ITS), translation elongation factor 1-α gene (TEF), RNA polymerase II second largest subunit (RPB2), major allergen Alt a 1 gene (Alt a 1), endopolygalacturonase gene (endoPG), anonymous gene region (OPA10-2) and glyceraldehyde 3-phos-phatedehydrogenase (GAPDH) were amplified and sequenced using primers ITS1/ITS4 (Peever et al. 2004), EF1-728F/EF1-986R (Carbone et al. 1999), RPB2-5F2/RPB2-5R (Sung et al. 2007), Alt-for/Alt-rev (Hong et al. 2005), EPG-specific/EPG-3b (Peever et al. 2004), OPA10-2R/OPA10-2L (Peever et al. 2004) and Gpd1/Gpd2 (Berbee et al. 1999) (GenBank accession no.MN046364, MW016001, MW016002, MW016003, MW016004, MW016005, MW016006). DNA sequences of TEF, RPB2, endoPG, OPA10-2 and GAPDH were 100% identical to those of A. tenuissima (MN256108, MK605866, KP789503, JQ859829 and MK683802), but ITS and Alt a 1 were 100% similarity with A. tenuissima (MN615420, JQ282277) and A. alternate (MT626589, KP123847). The ITS and Alt a 1 sequence did not distinguish A. tenuissima from the A. alternate complex. Maximum likelihood phylogenetic analyses were performed for the combined data set with TEF, RPB2, and endoPG using MEGA6 under the Tamura-Nei model (Kumar et al. 2016). The isolate Q1 clustered with type strain A. tenuissima CBS 918.96. The 20 celery plants of 4-7 leaf age were used test the pathogenicity of Q1, the ten plants were sprayed with 20ml of spore suspension (1×105 spores/ml), the control was sprayed with 20mL sterile water, which were placed in a growth chamber (25℃, a 14h light and 10h dark period, RH > 80%). Eight days after inoculation, 40% of the leaves formed lesions, which were consistent with the field observation,the control group was asymptomatic. The pathogen was reisolated from infected leaves to fulfill Koch's postulates. To our knowledge, this is the first report of A. tenuissima causing leaf spot on celery in China.


The productive and reproductive biology of flowering plants : VII. resource allocation and reproductive capacity in wild populations of Heloniopsis orientalis (Thunb.) C. Tanaka (Liliaceae).
The seasonal growth cycles and dry matter partitioning of Heloniopsis orientalis (Thunb.) C. Tanaka (Liliaceae) into its component organs throughout the year were studied and compared with one another in five selected populations at different altitudes in Toyama Prefecture, Japan (i.e., 100, 200, 900, 1,900, and 2,600 m above sea level). In addition, reproductive energy allociation (RA) and reproductive capacities of these five populations were critically analyzed in relation to environmental factors of each habitat.As a result, a remarkable cline in the growth cycle, dry matter allocation as well as significant reproductive characteristics including reproductive capacity and allocation was discovered along the environmental gradients (e.g., the length of the growing season) present as a result of different elevations. There is a clear negative correlation between the reproductive capacities and allocations in these five populations (r=0.9236, P<0.05), i.e., the populations of H. orientalis allocate increasingly greater energy to reproductive activities in response to the increase in elevation, but, on the contrary, show a marked decreasing tendency in reproductive capacity. Likewise, there is a tendency not only of increased energy allocation to total reproductive structures, but also toward producing a single propagule in successively harsher habitats. Specifically, the RA to total reproductive structures at the fruiting stage incrases from 9.97%, 13.03%, 15.25%, 23.06% to 25.52%, and also the relative amount of the energy invested to producing a single propagule increases from 1.000, 1.298, 2.496, 5.512 to 6.079 in response to an increase in elevation.


Congenic/consomic models of hypertension.
Human essential hypertension is a complex, multifactorial, quantitative trait under polygenic control. Despite major recent advances in genome sequencing and statistical tools, the genetic dissection of essential hypertension still provides a formidable challenge. Genetic models of essential hypertension such as the spontaneously hypertensive stroke-prone rat (SHRSP) provide the scientist with genetic homogeneity, not possible within a human population, to aid the search for causative genes. The principal strategy in the rat has been the identification of quantitative trait loci (QTL) responsible for blood-pressure regulation by genome-wide scanning. In this chapter we focus on congenic and consomic breeding strategies for the confirmation of QTL and the genetic dissection of the implicated regions.


Unique transcriptional profile of liver-resident memory CD8+ T cells induced by immunization with malaria sporozoites.
Sterile immunity against live Plasmodium infection can be achieved by immunization with radiation-attenuated sporozoites. This protection is known to be mediated in part by antigen-specific memory CD8(+) T cells, presumably those residing in the liver. We characterized and compared the transcriptional profile of parasite-specific memory CD8(+) T cells residing in the liver and spleen after immunization of mice with irradiated sporozoites. Microarray-based expression analysis of these memory CD8(+) T cells indicated that liver-resident memory cells display a distinct gene expression profile. We found major differences in the expression of immune function genes as well as genes involved in the cell cycle, cell trafficking, transcription and intracellular signaling. Importantly, the malaria parasite-induced liver-resident CD8(+) T cells display a transcriptional profile different to that described for CD8(+) T cells following other microbial challenges.


A Comparison of Probabilistic and Deterministic Match Strategies for Linking Prehospital and in-Hospital Stroke Registry Data.
Understanding and improving EMS stroke care requires linking data from both the prehospital and hospital settings. In the US, such data is collected in separate de-identified registries that cannot be directly linked due to lack of a common, unique patient identifier. In the absence of unique patient identifiers two common approaches to linking databases are deterministic matching, which uses combinations of non-unique matching variables to define matches, and probabilistic matching, which generates estimates of match probability based on the degree of similarity between records. This analysis seeks to compare these two approaches for matching EMS and stroke registry data. Stroke cases transported by EMS to Michigan hospitals participating in the Michigan Coverdell Acute Stroke Registry were linked to records from Michigan's EMS Information System (MI-EMSIS) between January 2018 and June 2019. Destination hospital, date-of-service, patient age, date-of-birth, and sex were used to perform deterministic and probabilistic linkages. Match rates and representativeness of the matched samples were compared between the two matching strategies. Multivariable logistic regression was used to identify characteristics associated with successful matching. During the 18-month study period there were 8,828 EMS transported confirmed stroke cases in the registry and 620,907 EMS transports to 38 Coverdell registry-participating hospitals. The probabilistic match linked 5985 (67.7%) strokes to EMS records; the deterministic match linked 4012 (45.5%). Within each strategy the characteristics of matched and unmatched cases were similar, with the exception that deterministically matched cases were less likely to be older than 89 (adjusted odds ratio [aOR]=0.3), white (aOR=0.8), and more likely to have subarachnoid hemorrhage (aOR=1.4) than unmatched cases. Probabilistic matching resulted in higher match rates and a more representative sample of EMS transported strokes, suggesting it may be superior in assessing EMS stroke care compared to a deterministic approach.


Impact of Aortic Valve Function and the Need for Aortic Valve Repair on Long-Term Outcomes of Valve-Sparing Aortic Root Replacement: 13-Year Experience of David Operation.
The best approach for aortic root disease remains controversial. Composite valve-graft conduit (CVG) replacement offers good results at short-term and long-term follow-up; on the other hand, valve-sparing aortic root replacement (VSARR) has proven to be an excellent treatment alternative. This study aimed to analyse the outcomes after VSARR and compare whether preoperative moderate or severe aortic regurgitation (AR) and or the need for aortic valve repair (AVR) during this procedure influenced survival and freedom from reoperation rates. From September 2005 to June 2018, 104 patients underwent VSARR using the reimplantation technique: 64% presented with preoperative moderate or severe AR, concomitant AVR was performed in 43.3%, Marfan syndrome was present in 16.3%, and 12.5% had a bicuspid aortic valve. Complete follow-up was obtained in 91% of the sample, echocardiographic results were available for 86% and the mean follow-up time was 1,893 days. In-hospital mortality was 2.9% and one death occurred 42 days after hospital discharge. In the latest echocardiographic assessment, 88.3% presented with mild AR or better. Freedom from reoperation at 8 years was 95.4%. There was no case of endocarditis and one patient had a stroke 2 years after the operation. There were no between-group differences in morbidity, mortality and complications during the follow-up. VSARR can be performed with low mortality rates and reasonable durability of the aortic valve. Neither moderate or severe AR nor the need for aortic valve repair during the procedure altered survival and freedom from reoperation.


A high-performance liquid chromatography method for determination of 4-aminopyridine in tissues and urine.
An analytical method was developed to measure 4-aminopyridine in tissues and urine to determine appropriate diagnostic samples in acute poisoning cases. Tissues from rats dosed with 4-aminopyridine were extracted with methylene chloride. Extracts were analyzed by high-performance liquid chromatography using an isocratic solvent system of acetonitrile and aqueous solution (15/85 v/v) consisting of 0.015 M sodium salt of l-heptane-sulfonic acid, 0.002 M tetramethylammonium bromide, and 0.01 M sodium dihydrogen phosphate adjusted to pH 3.0 with phosphoric acid. We concluded that suitable diagnostic samples for acute poisoning cases include stomach contents, kidney, liver, and urine.


The rabbit as an animal model for post-natal vitreous matrix differentiation and degeneration.
This study evaluates whether rabbits are a suitable animal model to study post-natal vitreous differentiation and degeneration. Human and rabbit eyes of various ages were studied by complementary anatomical techniques, light microscopy, and transmission electron microscopy. The global vitreous matrix organization is similar in human and rabbit eyes, lamellae are an important aspect thereof and show striking morphological changes with increasing age. In humans, liquefaction is more conspicuous than in rabbits but changes in matrix histology consistent with liquefaction can also be observed in the latter. Lamellar development is consistent with vitreous differentiation, while increasing liquefaction is consistent with matrix degeneration. At the anatomical and histological levels, human and rabbit vitreous matrices are sufficiently similar to make the rabbit a promising animal model for the study of the pathogenesis of vitreous matrix differentiation and degeneration in more detail.


Oxygen delivery to and use by primary porcine hepatocytes in the HepatAssist 2000 system for extracorporeal treatment of patients in end-stage liver failure.
A system for the extracorporeal treatment of end-stage liver failure patients has been developed and is now in clinical studies. This HepatAssist 2000 system consists of perfusing the plasma of the patient through a hollow-fiber bioreactor containing primary porcine hepatocytes. The goal of the system is to assist the patient while the liver regenerates or a suitable liver is found for transplantation. During the development of this system in the laboratory, an experimental protocol has been developed to simulate patient treatments. The protocol substitutes donor calf serum for patient plasma, but is otherwise similar to clinical conditions. The protocol uses a blood gas analyzer to monitor dissolved oxygen and carbon dioxide tensions at the entrance to and exit from the bioreactor. Samples are withdrawn using a standard 1-ml syringe and analyzed off-line immediately. The blood gas measurements have been used to verify that the oxygenator, placed immediately prior to the bioreactor in the plasma circuit, is properly sized to allow complete equilibration with feed gas and to maintain a physiologic level of carbon dioxide. The measurements were also used to calculate overall bioreactor oxygen consumption rates under various conditions. In one set of studies, composition of the feed gas to the oxygenator was varied from 20% to 70% oxygen, while maintaining 5% carbon dioxide and balance nitrogen. Rates of oxygen consumption and liver cell function (metabolism of diazepam) were unchanged, and no signs of oxygen toxicity were observed. In another set of studies, oxygen consumption increased linearly with number of hepatocytes used in the clinically used range of 5-10 billion hepatocytes, whether all hepatocytes were placed in one device or split between two devices operating in series or in parallel. In a third set of studies, oxygen consumption was a weak function of flowrate through the bioreactor but was highest at the clinically used recirculation rate of 400 ml/min. These data indicate that blood gas meters--readily available in hospital clinical laboratories--can be used to assist in the design of extracorporeal cell therapies, to monitor the effectiveness of system components, and to assess the effects of system modifications. These same measurements can be made noninvasively in the clinical setting to monitor oxygen consumption rates during patient treatment.


Protective effects of Thai silk sericins and their related mechanisms on UVA-induced phototoxicity and melanogenesis: Investigation in primary melanocyte cells using a proteomic approach.
UV radiation causes excess production of melanin as a result of hyperpigmentation and skin disorders. Silk sericin exhibited bioactivities to skin and inhibited UV-induced phototoxicity and melanogenesis in skin cells; however, the mechanism related to sericin against UV-induced melanogenesis has not been investigated. This study aimed to investigate the protective effects of Thai silk sericins against UVA-induced phototoxicity and melanogenesis and their related mechanisms. Thai silk sericins exhibited cytoprotective effects against UV-induced toxicity in human primary melanocytes by attenuation of cytotoxicity, intracellular ROS generation, and mitochondrial potential impairment. Pre- and post-treatment with sericin significantly inhibited melanin synthesis and tyrosinase activity against UVA exposure. In addition, sericin S2 could reduce the basal melanin content in zebrafish embryos. The proteomic analysis demonstrated that Thai silk sericins altered the protein expression in melanocytes especially proteins related to stress, inflammatory, cytokine stimulation, cell proliferation, and cell survival processes that contribute to cytoprotective effect and inhibitory effect on melanogenesis of sericin. Moreover, we demonstrated the novel mechanism of Thai silk sericins in inhibiting UVA-induced melanogenesis via increasing BMP4 expression in MAPK/ERK signaling pathway. These evidences support the potential use of Thai silk sericins in prevention of hyperpigmentation in skin disorders especially after UVA exposure.


An assessment of early Child Life Therapy pain and anxiety management: A prospective randomised controlled trial.
Burns remain extremely painful and distressing in young children. The consequences of poorly managed pain and anxiety can be life-long. Whilst Child Life Therapy (CLT) has been shown to be effective in many situations, few studies have looked at the effectiveness of CLT in regard to reducing pain and anxiety in children undergoing burn dressing changes. A prospective, randomised controlled trial was conducted, comparing CLT versus standard care in relation to pain and anxiety scores of children undergoing their initial burn dressing change. Pain and anxiety were assessed by an independent observer and questionnaires completed by the child, parent/caregiver and nursing staff. 50 subjects were recruited in each treatment group; median age 2.3 years (CLT) and 2.2 years (standard care). The median total body surface area (TBSA) burnt was 0.8% (CLT) and 0.5% (standard care). The majority were partial thickness dermal burns (88% CLT, 94% standard care). Rates of parent anxiety and pre-procedural child pain and anxiety were similar. Combined and scaled pain and anxiety scores in the CLT group were significantly less than in the standard treatment group (p=0.03). Whilst pain was significantly better in the CLT group (p=0.02), fear scores, wound outcomes and the need for skin grafting were not statistically different in either group. The presence of a Child Life Therapist, with their ability to adapt to the environment, the child and their family, significantly reduced the experience of pain during paediatric burn dressings.


Recognition and management of disseminated intravascular coagulation in horses.
This article reviews normal hemostasis in order to provide the reader with the basis for understanding the pathogenesis and manifestations (both clinical and laboratory) of disseminated intravascular coagulation (DIC) in horses. DIC is subsequently discussed. The diagnosis and treatment of DIC in horses are also described.


Giant midoesophageal diverticulum--case report and review of the literature.
Midoesophageal diverticula >4cm in size are a medical rarity with a dozen reported cases so far. Usually, midoesophageal diverticula tend to be of small size and asymptomatic. We herein present a case of a woman suffering from a midoesophageal diverticulum in a very large dimension--never described in the current literature before--who successfully underwent surgical resection via thoracotomy. If symptomatic, common symptoms are dysphagia and regurgitation. The risk of malignant transformation is low. Treatment of midoesophageal diverticula is based on size and symptoms of the patient. Asymptomatic or small diverticula detected during routine endoscopy can be followed-up without further therapy, whereas symptomatic or large diverticula should be treated surgically by resection. Myotomy should be performed if any motility disorder is evident. Open as well as minimal invasive approach by thoracoscopical surgery is both feasible.


A cost-effective, machine learning-based new unified risk-classification score (NU-CATS) for patients with endometrial cancer.
Treatment for endometrial cancer (EC) is increasingly guided by molecular risk classifications. Here, we aimed at using machine learning (ML) to incorporate clinical and molecular risk factors to optimize risk assessment. The Cancer Genome Atlas-Uterine Corpus Endometrial Carcinoma (n = 596), Memorial Sloan Kettering-Metastatic Events and Tropisms (n = 1315) and the American Association for Cancer Research Project Genomics Evidence Neoplasia Information Exchange (n = 4561) datasets were used to identify genetic alterations and clinicopathological features. Software packages including Keras, Pytorch, and Scikit Learn were tested to build artificial neural networks (ANNs) with a binary output as either intra-abdominal metastatic progression ('1') vs. non-metastatic ('0'). Black patients with EC have worse prognosis than White patients, adjusting for TP53 or POLE mutation status. Over 75% of Black patients carry TP53 mutations as compared to approximately 40% of White patients. Older age is associated with an increasing likelihood of TP53 mutation, high risk histology, and distant metastasis. For patients above age 70, 91% of Black and 60% of White EC patients carry TP53 mutations. A ML-based New Unified classifiCATion Score (NU-CATS) that incorporates age, race, histology, mismatch repair status, and TP53 mutation status showed 75% accuracy in prognosticating intra-abdominal progression. A higher NU-CATS is associated with an increasing risk of having positive pelvic or para-aortic lymph nodes and distant metastasis. NU-CATS was shown to outperform Leiden/TransPORTEC model for estimating risk of FIGO Stage I/II disease progression and survival in Black EC patients. The NU-CATS, a ML-based, cost-effective algorithm, incorporates diverse clinicopathologic and molecular variables of EC and yields superior prognostication of the risk of nodal involvement, distant metastasis, disease progression, and overall survival.


Association of nocturia of self-report with estimated glomerular filtration rate: a cross-sectional study from the NHANES 2005-2018.
Nocturia is a manifestation of systemic diseases, in which chronic kidney disease (CKD) is an independent predictor of nocturia due to its osmotic diuretic mechanism. However, to our knowledge, previous studies have not examined the association between nocturia and estimated glomerular filtration rate (eGFR). The purpose of this study was to assess the association between nocturia exposure and eGFR in the general US population. This study presents a cross-sectional analysis of the general US population enrolled in the National Health and Nutrition Examination Survey (NHANES) from 2005 to 2018. To account for potential confounding factors, linear regression analysis was conducted to investigate the association between nocturia and eGFR. Stratified analyses and interaction tests were employed to examine the variables of interest. Additionally, sensitivity analyses were conducted across diverse populations. A total of 12,265 individuals were included in the study. After controlling for confounding factors, the results of the linear regression analysis indicated that a single increase in nocturnal voiding frequency was associated with a decrease in eGFR by 2.0 mL/min/1.73 m2. In comparison to individuals with a nocturnal urinary frequency of 0, those who voided 1, 2, 3, 4, and ≥ 5 times at night experienced a decrease in eGFR by 3.1, 5.4, 6.4, 8.6 and 4.0 mL/min/1.73 m2, respectively. Nocturia was found to be associated with a decreased eGFR of 4 mL/min/1.73 m2 when compared to individuals without nocturia. The sensitivity analysis yielded consistent findings regarding the association between nocturia and eGFR in both CKD and non-CKD populations, as well as in hypertensive and non-hypertensive populations. Nevertheless, inconsistent conclusions were observed across various prognostic risk populations within the CKD context. The presence of nocturia and heightened frequency of nocturnal urination have been found to be associated with a decline in eGFR.