HIF-1α inhibitor PX-478 preserves pancreatic β cell function in diabetes.
During progression of type 2 diabetes, pancreatic β cells are subjected to sustained metabolic overload. We postulated that this state mediates a hypoxic phenotype driven by hypoxia-inducible factor-1α (HIF-1α) and that treatment with the HIF-1α inhibitor PX-478 would improve β cell function. Our studies showed that the HIF-1α protein was present in pancreatic β cells of diabetic mouse models. In mouse islets with high glucose metabolism, the emergence of intracellular Ca2+ oscillations at low glucose concentration and the abnormally high basal release of insulin were suppressed by treatment with the HIF-1α inhibitor PX-478, indicating improvement of β cell function. Treatment of db/db mice with PX-478 prevented the rise of glycemia and diabetes progression by maintenance of elevated plasma insulin concentration. In streptozotocin-induced diabetic mice, PX-478 improved the recovery of glucose homeostasis. Islets isolated from these mice showed hallmarks of improved β cell function including elevation of insulin content, increased expression of genes involved in β cell function and maturity, inhibition of dedifferentiation markers, and formation of mature insulin granules. In response to PX-478 treatment, human islet organoids chronically exposed to high glucose presented improved stimulation index of glucose-induced insulin secretion. These results suggest that the HIF-1α inhibitor PX-478 has the potential to act as an antidiabetic therapeutic agent that preserves β cell function under metabolic overload.


Deleterious effect of cyclosporins on the ischemic kidney in the rat and the protection by the calcium antagonist verapamil.
Cyclosporine A (CsA) nephrotoxicity has been suggested to be aggravated in the presence of ischemia, as occurs after renal transplantation. Cyclosporine G (CsG) may be less nephrotoxic than CsA. This study evaluated in the rat (1) the effect of CsA and CsG on blood flow and the function of the kidney subjected to 60 min of warm ischemia and (2) the protective effect of the calcium antagonist verapamil (VP). After left nephrectomy, ischemia was induced in the right kidney by the clamping of the kidney pedicle for 60 min, which resulted in a significant increase in serum creatinine (SCr) to 2.30 +/- 0.25 mg/dL by Day 1 with 25% mortality by Day 7. The administration of CsA or CsG (20 mg/kg i.v. daily for 7 days) after 60 min of renal ischemia significantly increased SCr and mortality compared with ischemia alone. In another set of experiments, 60 min of warm ischemia was applied to the right kidney and RBF was measured in both kidneys with a laser Doppler flowmeter. Blood flow in the ischemic kidney returned to the preischemic level by 15 min after the removal of the vascular clamp in the control animals. In contrast, in animals treated with CsA, a significant decrease in RBF was seen in both kidneys; however, blood flow in the ischemic kidney was significantly lower than that in the nonischemic kidney. CsG also decreased RBF in both kidneys, although in the left (nonischemic) kidney, RBF remained significantly higher with CsG than with CsA.(ABSTRACT TRUNCATED AT 250 WORDS)


Plasmoacanthoma.
Plasmoacanthoma is an extremely rare verrucous tumor located on periorificial regions characterized by dense dermal plasmacytic infiltrates. Some authors classify it as a form of reactive plasma cell proliferation which represents a heterogeneous spectrum of mucocutaneous disorders. These plasma cell proliferations have been considered to be a benign immunologic inflammatory reaction to known or unknown stimuli. However, the etiology of plasmoacanthoma remains highly speculative. We report the case of a 40-year-old woman who presented with a lobulated warty lesion affecting the lower lip. Biopsy from the lesion was compatible with plasmoacanthoma, which remains an underreported disease in the dermatology literature.


Establishing Clinical Pharmacy Services With Prescribing Privileges in a Federally Qualified Health Center Primary Care Clinic.
To describe the process and cost of establishing clinical pharmacy services with prescribing privileges in a federally qualified health center (FQHC) primary care clinic. The primary care clinic was located in a low-income area of Southern California and served patients with Medicaid and Medicare. The primary care clinic had preventive medicine and family medicine physicians, a family medicine residency program, behavioral health services, and a registered dietician. New clinical pharmacy services were established at this FQHC primary care clinic. The medication assistance program was a stepping stone to establish rapport with the physicians. Credentialing and privileging was implemented for clinical pharmacists. An open protocol collaborative practice agreement was developed to allow clinical pharmacists to manage ambulatory patients. From August 2014 to June 2015, the clinical pharmacist interacted with 392 patients and spent 336 hours educating patients and providing disease state management. The pharmacist also provided consults to residents and providers. Diabetic patients made up 76% of all clinical pharmacy encounters. There were 86 face-to-face clinical pharmacy appointments with the pharmacist. The average time for clinical pharmacy appointments was 77 minutes. By describing ways to develop rapport with providers, how to credential and privilege pharmacists, and explain resources and costs of setting up a service, the hope is that more clinical pharmacists will be able to incorporate into independent or FQHC primary care clinics for improved management of ambulatory patients.


Comparative Efficacy of Local Steroid and Therapeutic Ultrasound for Quicker Functional Improvement in Tennis Elbow.
Lateral epicondylitis or Tennis elbow is very common repetitive stress injury or cumulative trauma disorder among active professionals and tennis players. Local steroid injection can provide quick functional improvement as sought by most of the patients. Therapeutic ultrasound and eccentric exercises can also improve the function. This study was carried out to see the comparative efficacy of these two modalities of treatments in terms of quicker functional improvement and this randomized interventional clinical trial was carried out in a private rehabilitation hospital in Dhaka. Patients attended the authors private chamber from January 2013 to December 2013 with clinical diagnosis of tennis elbow were included. Eighty adult patients diagnosed as lateral epicondylitis or tennis elbow were randomly divided into two groups. Group A received local steroid injection (triamcinolone acetate) and group B received therapeutic ultrasound for two weeks. Both the groups received eccentric exercises and advised to avoid provocative activities like twisting and lifting activities. Pain, tenderness and functional improvements were measured in traditional methods like visual analogue scale (VAS) and tenderness grade 0 to 4. Data were analyzed by SPSS. Most of the patients were female. None of them were tennis player. Younger housewives were common sufferer. Almost all patients in group A had very little or no pain or tenderness and no difficulty in lifting object at one week and two weeks. VAS was 13.62±6.503 and 4.15±5.517 respectively. Group B patients had mild improvement of pain and function at one week and moderate improvement at two weeks. VAS was 52.57±7.80 and 25.57±5.392 respectively. Similarly tenderness' were markedly improved in Group A at one and two weeks. They were 0.68±0.616 and 0.02±0.405 respectively for Group A and 1.60±0.553 and 1.14±0.335 respectively for Group B. Results were statistically significant in both groups at the end of two weeks. Local corticosteroid injections at extensor origin in tennis elbow patients are found to have better relief of pain and tenderness and quicker functional improvement than therapeutic ultrasound treatment. None of the patients were followed up for long term efficacy in either group.


Spontaneous release of leukocyte migration inhibition factor (LMIF) by lymphocytes in chronic lymphatic leukemia patients.
Leukocyte migration inhibition factor (LMIF) activity expressed as a migration index was studied in sera of 13 chronic lymphatic leukemia (CLL) patients and 14 healthy subjects. LMIF activity was present in 92.8% of CLL patients and 14% of controls. The LMIF activity of lymphocyte culture supernatants in the same patients and 7 controls was also studied. A LMIF-like activity was found both, in the sera and phytohemagglutinin (PHA) unstimulated cell supernatants in 9 of 13 patients (69.2%). These results demonstrate the ability of malignant B cells to release LMIF or LMIF-like factors spontaneously.


A serine/threonine protein kinase gene isolated by an in vivo binding procedure using the Arabidopsis floral homeotic gene product, AGAMOUS.
During the course of characterizing fragments bound to an Arabidopsis floral protein AGAMOUS in vivo, a gene encoding a putative serine/threonine protein kinase was found on one of the fragments. The deduced 426 amino acid residues of the gene, named APK2a, are 65% identical to a previously reported Arabidopsis serine/threonine protein kinase, APK1a. The gene is composed of 6 exons and maps at 10 cM from the upper end of chromosome 1. Northern hybridization experiments indicated that the gene is strongly expressed in leaves, moderately in roots, and very weakly in flowers. Further in situ analysis of the expression in floral buds showed that the APK2a gene is expressed at pedicels, is not expressed at the floral organ primordia of wild type floral buds, but is moderately expressed in the floral organ primordia of the agamous mutant. In vitro binding assay suggest that the AGAMOUS protein binds to a sequence similar to, but different from, the known MADS-binding consensus sequences, the CArG box, located 3' downstream of the APK2a gene. These results suggest that APK2a expression is negatively regulated by the AG protein. A close homologue of the APK2a gene, named APK2b, was also isolated from the Arabidopsis cDNA library. The expression pattern of the APK2b gene differs from that of APK2a. It is strongly expressed in leaves, moderately in flowers, and weakly in roots.


Characterization of Industry 4.0 Lean Management Problem-Solving Behavioral Patterns Using EEG Sensors and Deep Learning.
Industry 4.0 leaders solve problems all of the time. Successful problem-solving behavioral pattern choice determines organizational and personal success, therefore a proper understanding of the problem-solving-related neurological dynamics is sure to help increase business performance. The purpose of this paper is two-fold: first, to discover relevant neurological characteristics of problem-solving behavioral patterns, and second, to conduct a characterization of two problem-solving behavioral patterns with the aid of deep-learning architectures. This is done by combining electroencephalographic non-invasive sensors that capture process owners' brain activity signals and a deep-learning soft sensor that performs an accurate characterization of such signals with an accuracy rate of over 99% in the presented case-study dataset. As a result, the deep-learning characterization of lean management (LM) problem-solving behavioral patterns is expected to help Industry 4.0 leaders in their choice of adequate manufacturing systems and their related problem-solving methods in their future pursuit of strategic organizational goals.


Shifting social-ecological fire regimes explain increasing structure loss from Western wildfires.
Structure loss is an acute, costly impact of the wildfire crisis in the western conterminous United States ("West"), motivating the need to understand recent trends and causes. We document a 246% rise in West-wide structure loss from wildfires between 1999-2009 and 2010-2020, driven strongly by events in 2017, 2018, and 2020. Increased structure loss was not due to increased area burned alone. Wildfires became significantly more destructive, with a 160% higher structure-loss rate (loss/kha burned) over the past decade. Structure loss was driven primarily by wildfires from unplanned human-related ignitions (e.g. backyard burning, power lines, etc.), which accounted for 76% of all structure loss and resulted in 10 times more structures destroyed per unit area burned compared with lightning-ignited fires. Annual structure loss was well explained by area burned from human-related ignitions, while decadal structure loss was explained by state-level structure abundance in flammable vegetation. Both predictors increased over recent decades and likely interacted with increased fuel aridity to drive structure-loss trends. While states are diverse in patterns and trends, nearly all experienced more burning from human-related ignitions and/or higher structure-loss rates, particularly California, Washington, and Oregon. Our findings highlight how fire regimes-characteristics of fire over space and time-are fundamentally social-ecological phenomena. By resolving the diversity of Western fire regimes, our work informs regionally appropriate mitigation and adaptation strategies. With millions of structures with high fire risk, reducing human-related ignitions and rethinking how we build are critical for preventing future wildfire disasters.


Lactobacillus insicii sp. nov., isolated from fermented raw meat.
The analysis of the bacterial microbiota of retain samples of pork salami revealed an isolate (strain TMW 1.2011T) that could neither be assigned to typical genera of starter organisms nor to any other known meat-associated species. Cells were Gram-stain-positive, short, straight rods occurring singly, in pairs or short chains. Phylogenetic analysis of the 16S rRNA gene sequence and specific phenotypic characteristics showed that strain TMW 1.2011T belonged to the phylogenetic Lactobacillus alimentarius group, and the closest neighbours were Lactobacillus nodensis JCM 14932T (97.8 % 16S rRNA gene sequence similarity), Lactobacillus tucceti DSM 20183T (97.4 %), 'Lactobacillus ginsenosidimutans' EMML 3041 (97.3 %), Lactobacillus versmoldensis DSM 14857T (96.9 %) and Lactobacillus furfuricola JCM 18764T (97.2 %). Similarities using partial gene sequences of the alternative chronometers pheS, dnaK and rpoA also support these relationships. DNA-DNA relatedness between the novel isolate and L. nodensis JCM 14932T, L. versmoldensis DSM 14857T and L. tucceti DSM 20183T, L. furfuricola JCM 18764T and 'L. ginsenosidimutans' EMML 3041 were below 70 % and the DNA G+C content was 36.3 mol%. The cell-wall peptidoglycan type is l-Lys-Gly-d-Asp. Based on phylogenetic, chemotaxonomic and physiological evidence, strain TMW 1.2011T represents a novel species of the genus Lactobacillus, for which the name Lactobacillus insicii sp. nov. is proposed. The type strain is TMW 1.2011T ( = CECT 8802T = DSM 29801T).


Combined Multivariate Statistical Techniques and Water Quality Index (WQI) to Evaluate Spatial Variation in Water Quality.
In present study, Water Quality Index (WQI) has been assessed of the Rawal Lake which is a major source of drinking water for people in the Federal Capital, Islamabad, and its adjacent city Rawalpindi in Pakistan. For this, the principal component analysis (PCA) and WQI were applied as an integrated approach to quantitatively explore difference based on spatial variation in 11 water quality parameters of the five major feeding tributaries of the Rawal Lake, Pakistan. The results of temperature in water, total dissolved solids, pH, electrical conductivity, chlorides and sulfates were well within the allowable World Health Organisation's (WHO) limits. However, the heavy metals like cadmium and lead were above permissible limits by the WHO in tributaries of Bari Imam and Rumli. Moreover, this has been proven by the Pearson correlation which suggested strong positive correlation (0.910*) between lead and cadmium. The results of present study were subjected to statistical analysis, i.e., PCA which gave three major factors contributing 96.5% of the total variance. For factor 1, pH, TDS, alkalinity, chlorides, sulfates and zinc have highest factor loading values (>0.60) and presented that these parameters were among the most significant parameters of first factor. As per the WQI results, the water was categorised in two major classes indicating that water of Bari Imam and Rumli is highly contaminated with heavy metals and totally unsuitable for drinking purposes. Based on the results of the present study, it is suggested to make heavy metals consideration as an integrated component in future planning for maintaining water quality of the Rawal Lake and its tributaries.


Female vs. Male Ampelmännchen-Gender-Specific Reaction Times to Male and Female Traffic Light Figures.
Traffic signs are important visual guiding signals for the safe navigation through complex road traffic. Interestingly, there is little variation in the traffic signs for cars around the world. However, remarkable variation exists for pedestrian traffic signs. Following up from an earlier study, we investigated the visual efficacy of female vs. male German Ampelmännchen pedestrian traffic signs. In a Stroop-like test, 30 subjects were presented with female and male go and no-go traffic light figures that were shown either in the corresponding or opposing color. Subjects had to indicate, based either solely on the form or the color of the figure, whether they were allowed to go. Accuracy and response times across all subjects did not differ for the female vs. male signs, indicating that Ampelfrau and Ampelmann signs have equal visual efficacy. However, subjects responded faster to signs of their own vs. the opposite gender. This preference for signs of one's own gender is in accordance with effects in social psychology described by social learning theory. An introduction of such novel traffic lights may, thus, contribute to higher compliance with the traffic sign signals.


Cloning, DNA sequence, functional analysis and transcriptional regulation of the genes encoding dipicolinic acid synthetase required for sporulation in Bacillus subtilis.
Dipicolinic acid (DPA) is a small polar molecule that accumulates to high concentrations in bacterial endospores, and is thought to play a role in spore heat resistance, or the maintenance of heat resistance. Previous work has shown that mutations in the spoVF locus of Bacillus subtilis prevent the formation of DPA, and give rise to heat-sensitive spores. Addition of exogenous DPA during spore development led to the restoration of heat resistance. This suggested that the spoVF locus encoded dipicolinic acid synthetase, the enzyme thought to catalyse the single reaction needed to synthesise DPA from dihydroxydipicolinic acid, an intermediate in the lysine biosynthetic pathway. We have now cloned and sequenced the spoVF locus of Bacillus subtilis and show that it comprises two coordinately regulated genes, now designated dpaA and dpaB. Expression of fragments of the dpa operon in Escherichia coli has shown that the two gene products together specify DPA synthetase activity. The promoter of the dpa operon, which lies just upstream of the first gene, has been identified by primer extension analysis. Sequences in this region show strong sequence similarity to several promoters recognized by the sigma K form of RNA polymerase. Transcription from this promoter was detected four hours after the onset of sporulation, at about the same time that sigma K activity is known to appear. Furthermore, transcription was abolished by mutations in a series of genes that are known to be required for the synthesis of active sigma K. These results are in accordance with previous work indicating that DPA synthetase activity was present only during the late stages of sporulation and specifically in the mother cell compartment. Transcription was enhanced by a gerE mutation, indicating that, like the previously described cotA gene, spoVF is negatively regulated by GerE. The mother-cell-specific synthesis of an enzyme responsible for a compound that accumulates to high concentrations in the prespore raises interesting questions about intercellular transport mechanisms.


Near-Infrared Coloring via a Contrast-Preserving Mapping Model.
Near-infrared gray images captured along with corresponding visible color images have recently proven useful for image restoration and classification. This paper introduces a new coloring method to add colors to near-infrared gray images based on a contrast-preserving mapping model. A naive coloring method directly adds the colors from the visible color image to the near-infrared gray image. However, this method results in an unrealistic image because of the discrepancies in the brightness and image structure between the captured near-infrared gray image and the visible color image. To solve the discrepancy problem, first, we present a new contrast-preserving mapping model to create a new near-infrared gray image with a similar appearance in the luminance plane to the visible color image, while preserving the contrast and details of the captured near-infrared gray image. Then, we develop a method to derive realistic colors that can be added to the newly created near-infrared gray image based on the proposed contrast-preserving mapping model. Experimental results show that the proposed new method not only preserves the local contrast and details of the captured near-infrared gray image, but also transfers the realistic colors from the visible color image to the newly created near-infrared gray image. It is also shown that the proposed near-infrared coloring can be used effectively for noise and haze removal, as well as local contrast enhancement.


Reaching Consensus on Outcomes for Successful Cannulation of an Arteriovenous Fistula: Patient and Healthcare Provider Perspectives.
There is a lack of consensus on what constitutes successful arteriovenous fistula (AVF) cannulation. The purpose of this study was to describe outcomes of successful cannulation of the AVF from both patient and healthcare provider perspectives. This was a mixed method study. Results reflecting the patient's description of success through interviews were reported previously. A sample of nursing and physician experts in vascular access completed a one-time survey. Results from healthcare providers suggest they consider cannulator ability to assess the AVF, knowledge of AVF anatomy, and patient-centered care as most important to cannulation success. Patient comfort, patient-centered care, and available support staff (i.e., expert cannulators) were perceived by both patients and healthcare provider groups as contributing to success. Strategies that promote patient comfort, patient-centered care, and having access to cannulation experts have the potential to improve cannulation outcomes.


A Case of Acquired Hemophilia A in a Patient with Exposure to COVID-19.
Acquired Hemophilia A (AHA) is a very rare autoimmune condition involving immune-mediated depletion of Factor VIII, resulting in spontaneous hemorrhage. Failure to recognize AHA as a possible etiology of hemorrhage can result in delayed diagnosis and treatment. The COVID-19 pandemic has given rise to several hematologic conditions and complications, with a rare manifestation being Acquired Hemophilia A (AHA). An interesting case of AHA following SARS-CoV-2 infection is described, along with the treatment approach and a brief review of several cases describing AHA in association with COVID-19.


Excited-State Deactivation of Branched Phthalocyanine Compounds.
The excited-state relaxation dynamics and chromophore interactions in two phthalocyanine compounds (bis- and trisphthalocyanines) are studied by using steady-state and femtosecond transient absorption spectral measurements, where the excited-state energy-transfer mechanism is explored. By exciting phthalocyanine compounds to their second electronically excited states and probing the subsequent relaxation dynamics, a multitude of deactivation pathways are identified. The transient absorption spectra show the relaxation pathway from the exciton state to excimer state and then back to the ground state in bisphthalocyanine (bis-Pc). In trisphthalocyanine (tris-Pc), the monomeric and dimeric subunits are excited and the excitation energy transfers from the monomeric vibrationally hot S1 state to the exciton state of a pre-associated dimer, with subsequent relaxation to the ground state through the excimer state. The theoretical calculations and steady-state spectra also show a face-to-face conformation in bis-Pc, whereas in tris-Pc, two of the three phthalocyanine branches form a pre-associated face-to-face dimeric conformation with the third one acting as a monomeric unit; this is consistent with the results of the transient absorption experiments from the perspective of molecular structure. The detailed structure-property relationships in phthalocyanine compounds is useful for exploring the function of molecular aggregates in energy migration of natural photosynthesis systems.


Development of Osteoarthritis in Adults With Type 2 Diabetes Treated With Metformin vs a Sulfonylurea.
Metformin may have a protective association against developing osteoarthritis (OA), but robust epidemiological data are lacking. To determine the risk of OA and joint replacement in individuals with type 2 diabetes treated with metformin compared with a sulfonylurea. This retrospective cohort study used claims data from the Optum deidentified Clinformatics Data Mart Database between December 2003 and December 2019. Participants included individuals aged 40 years or older with at least 1 year of continuous enrollment and type 2 diabetes. Individuals with type 1 diabetes or a prior diagnosis of OA, inflammatory arthritis, or joint replacement were excluded. Time-conditional propensity score matching was conducted using age, sex, race, Charlson comorbidity score, and treatment duration to create a prevalent new-user cohort. Data were analyzed from April to December 2021. Treatment with metformin or a sulfonylurea. The outcomes of interest were incident OA and joint replacement. Cox proportional hazard models were used to calculate adjusted hazard ratios (aHRs) of incident OA and joint replacement. In a sensitivity analysis, individuals only ever treated with metformin were compared with individuals only ever treated with a sulfonylurea, allowing for longer-term follow up of the outcome (even after stopping the medication of interest). After time-conditional propensity score matching, the metformin and control groups each included 20 937 individuals (mean [SD] age 62.0 [11.5] years; 24 379 [58.2%] males). In the adjusted analysis, the risk of developing OA was reduced by 24% for individuals treated with metformin compared with a sulfonylurea (aHR, 0.76; 95% CI, 0.68-0.85; P < .001), but there was no significant difference for risk of joint replacement (aHR, 0.80; 95% CI, 0.50-1.27; P = .34). In the sensitivity analysis, the risk of developing OA remained lower in individuals treated with metformin compared with a sulfonylurea (aHR, 0.77; 95% CI, 0.65-0.90; P < .001) and the risk of joint replacement remained not statistically significant (aHR, 1.04; 95% CI, 0.60-1.82; P = .89). In this cohort study of individuals with diabetes, metformin treatment was associated with a significant reduction in the risk of developing OA compared with sulfonylurea treatment. These results further support preclinical and observational data that suggest metformin may have a protective association against the development of OA; future interventional studies with metformin for the treatment or prevention of OA should be considered.


Involvement of fatty acid pathways and cortical interaction of the pronuclear complex in Caenorhabditis elegans embryonic polarity.
Cell polarity is essential for many decisions made during development. While investigation of polarity-specific factors has yielded great insights into the polarization process, little is known on how these polarity-specific factors link to the basic cellular mechanisms that function in non-polarity aspects of the cell. To better understand the mechanisms that establish embryonic polarity, we investigated genes required for polarity in the one-cell C. elegans embryo that are also required for other non-polarity functions. This has led to the identification of the Pod-class of mutants that are characterized by osmosensitive embryos and defects in anterior-posterior polarity. Mutation in either of two loci of this class, emb-8 and pod-2, disrupts embryonic polarization and results in osmotically-sensitive embryos. Loss of emb-8, a previously uncharacterized polarity gene, causes mislocalization of PAR-3 and PAR-2 that molecularly mark the anterior and posterior cortices. emb-8 encodes NADPH-cytochrome P450 reductase, a protein supplying electrons to cytochrome P450-family enzymes, some of which catalyze fatty acid modifications. Cloning of the previously characterized polarity gene pod-2 reveals it encodes acetyl-CoA carboxylase, an enzyme that catalyzes the first step in de novo fatty acid synthesis. Depletion of fatty acid synthase, the next enzyme in the biosynthetic pathway, by RNA-interference (RNAi) also causes similar loss of one-cell polarity. Furthermore, pod-2 polarity defects can be rescued by addition of exogenous fatty acids. By following the behavior of the pronucleus in emb-8 and pod-2 mutant embryos, we demonstrate that loss of polarity correlates with impaired interaction between the pronucleus-centrosome complex and the posterior cortex. The characterization of emb-8 and pod-2 mutant embryos suggests that the pronucleus-centrosome complex interaction with the cortex plays a direct role in establishing polarity and that fatty acid pathways are important for this polarizing event.


Influence of body mass index and age on day-of-surgery discharge, prolonged admission, and 90-day readmission after fast-track unicompartmental knee arthroplasty.
Background and purpose - The indications for unicompartmental knee arthroplasty (UKA) have become less restrictive and, today, high age and high BMI are not considered contraindications by many surgeons. While the influence of these patient characteristics on total knee arthroplasty is well documented, evidence on UKA is lacking. We investigated the effect of BMI and age on day of surgery (DOS) discharge, prolonged admission, and 90-day readmission following UKA surgery.Patients and methods - This retrospective cohort study included 3,897 UKA patients operated on between 2010 and 2018 in 8 fast-track arthroplasty centers. Patients were divided into 5 BMI groups and 5 age groups. Differences between groups in the occurrence of DOS discharge, prolonged admission > 2 days, and 90-day readmission was investigated using a chi-square test and mixed-effect models adjusted for patient characteristics using surgical center as a random effect.Results - Median LOS was 1 day. DOS discharge was achieved in 26% of patients with no statistically significant differences between BMI groups. DOS discharge was less likely in UKA patients aged > 70 years (age 71-80; odds ratio [OR] 0.7 [95% CI 0.6-0.9]). Prolonged admission was not affected by BMI or age in the adjusted analysis. 90-day readmission was more likely in patients with BMI > 35 (OR 1.9 [CI 1.1-3.1]) and patients aged 71-80 (OR 1.5 [CI 1.1-2.1]).Interpretation - Age > 70 years decreased the likelihood of DOS discharge after UKA. High BMI as well as advanced age increased the likelihood of 90-day readmission. This should be noted by surgeons operating on patients with high BMI and age.


Sustained white cell cytokine activation in idiopathic dilated cardiomyopathy despite haemodynamic improvement with medical therapy.
Cytokine production in idiopathic dilated cardiomyopathy (IDC) may depend on neurohumoral stimulation, haemodynamic impairment or auto-antibody production. We aimed to ascertain the impact of haemodynamic improvements with standard medical therapy and neurohumoral blockade on white cell tumour necrosis factor- alpha (TNF-alpha ) production in patients with IDC. Twenty-seven patients with IDC and NYHA class I to IV heart failure but without evidence of oedema, reduced peripheral perfusion, or elevated plasma endotoxin concentrations were evaluated for indicators of cytokine activation. Plasma TNF-alpha concentrations were raised (p < 0.001) in patients prior to commencement of medical therapy as compared to controls (n = 27). In addition, endotoxin-free cultured whole blood TNF-alpha production was enhanced (p < 0.02) in the patients. Although plasma TNF-alpha tended to decrease, excessive whole blood TNF-alpha production remained unaltered following marked improvements in haemodynamics and functional class (increase in absolute left ventricular ejection fraction = 8.7 +/- 2.6%, p < 0.01, 37% in NYHA functional class I after therapy) with six to 12 months of medical therapy (diuretic, angiotensin converting enzyme inhibitor and beta-blocker). Against a role for neurohumoral substances in promoting excessive white cell TNF-alpha synthesis the angiotensin II receptor antagonist, losartan, failed to modulate white cell TNF- alpha production in patients with IDC. We concluded that white cell TNF-alpha overproduction is sustained in patients with IDC despite haemodynamic improvement with standard medical therapy and blockade of angiotensin II receptors. These data suggest that mechanisms other than haemodynamic impairment and neurohumoral activation contribute to excess white cell TNF-alpha production in IDC.


Integration of ganglioside GT1b receptor into DPPE and DPPC phospholipid monolayers: an X-ray reflectivity and grazing-incidence diffraction study.
Using synchrotron grazing-incidence x-ray diffraction (GIXD) and reflectivity, the in-plane and out-of-plane structures of mixed-ganglioside GT(1b)-phospholipid monolayers were investigated at the air-liquid interface and compared with monolayers of the pure components. The receptor GT(1b) is involved in the binding of lectins and toxins, including botulinum neurotoxin, to cell membranes. Monolayers composed of 20 mol % ganglioside GT(1b), the phospholipid dipalmitoyl phosphatidylethanolamine (DPPE), and the phospholipid dipalmitoyl phosphatidylcholine (DPPC) were studied in the gel phase at 23 degrees C and at surface pressures of 20 and 40 mN/m, and at pH 7.4 and 5. Under these conditions, the two components did not phase-separate, and no evidence of domain formation was observed. The x-ray scattering measurements revealed that GT(1b) was intercalated within the host DPPE/DPPC monolayers, and slightly expanded DPPE but condensed the DPPC matrix. The oligosaccharide headgroups extended normally from the monolayer surfaces into the subphase. This study demonstrated that these monolayers can serve as platforms for investigating toxin membrane binding and penetration.


Occurrence of nuclear beta(II)-tubulin in cultured cells.
Microtubules are cylindrical organelles that play critical roles in cell division. Their subunit protein, tubulin, is a target for various antitumor drugs. Tubulin exists as various forms, known as isotypes. In most normal cells, tubulin occurs only in the cytosol and not in the nucleus. However, we have recently reported the finding of the beta(II) isotype of tubulin in the nuclei of cultured rat kidney mesangial cells. Mesangial cells, unlike most normal cell lines, have the ability to proliferate rapidly in culture. In efforts to determine whether nuclear beta(II)-tubulin occurred in other cell lines, we examined the distribution of the beta(I), beta(II), and beta(IV) mammalian tubulin isotypes in a variety of normal and cancer human cell lines by immunofluorescence microscopy. We have found that, in the normal cell lines, all three isotypes are present only in the cytoplasm. However, the beta(II) isotype of tubulin is located not only in the cytoplasm, but also in the nuclei of the following cell lines: LNCaP prostate carcinoma, MCF-7, MDA-MB-231, MDA-MB-435, and Calc18 breast carcinoma, C6 and T98G glioma, and HeLa cells. In contrast, the beta(I) and beta(IV) isotypes, which are also synthesized in cancer cells, are not localized to the nucleus but are restricted to the cytoplasm. We have also seen beta(II) in breast cancer excisions. In most of these cells, beta(II) appears to be concentrated in the nucleoli. These results suggest that transformation may lead to localization of beta(II)-tubulin in cell nuclei, serving an as yet unknown function, and that nuclear beta(II) may be a useful marker for detection of tumor cells.


Expression of growth factor/receptor genes in postthymic T cell malignancies.
This study was undertaken to explain the molecular basis for the diverse pathology and clinical behavior of postthymic T cell malignancies. Total cellular RNAs were extracted from four HTLV-1 positive and ten HTLV-1-negative T cell lymphomas and cell lines, and investigated for homology with cloned DNA probes specific for interleukin-2 (IL-2), IL-2 receptor (IL-2R), transforming growth factor beta (TGF-beta), platelet-derived growth factor (PDGF), and epidermal growth factor receptor (EGF-R). Tumor cells associated with clinically high grade HTLV-1-positive adult T cell leukemia (ATL) and large cell morphology (T immunoblastic lymphomas) were found to have higher levels of expression of IL-2 and TGF-beta genes than low grade T cell neoplasms (mycosis fungoides and Sezary's syndrome). High expression of IL-2R gene was restricted to Ki-1-positive lymphomas and to one ATL. Cell lines corresponding to the advanced stage of a cutaneous T cell lymphoma (CTCL) showed enhanced expression of PDGF. Therefore, high grade T cell malignancies had consistently elevated expression of growth factor/receptor (GF/R) genes. Expression of EGF-R was negligible in all T cell malignancies. An inverse relationship was found between the expression of T cell antigen receptor (differentiation antigen) and GF/R (activation antigen) genes, accounting for the frequent aberrant expression of T cell antigens in high grade T cell lymphomas. The results suggest that post-thymic T cell malignancies derived from activated T cells produce and secrete GF, conferring a growth advantage on neoplastic T cells, and correlating well with their histologic subtype and clinical behavior.


Heterologous Seminal Plasma Reduces the Intracellular Calcium and Sperm Viability of Cryopreserved Stallion Spermatozoa.
Despite the vital role of seminal plasma (SP) in maintaining sperm function and aiding gamete interaction in many species, SP is usually removed before cryopreservation of stallion sperm to improve cryosurvival of sperm. The present study assessed if the vital sperm functional parameters of genetically superior stallions producing poor quality semen can be enhanced by the supplementation of heterologous SP from the stallion producing high quality semen. Spermatozoa from poor quality semen producing stallions were divided into three aliquots: two aliquots were supplemented with SP obtained from good quality semen producing stallions at the rate of 20% and 30%, respectively, whereas the third aliquot remained as control (0% SP) and incubated at 37°C for 30 minutes. Sperm membrane integrity, mitochondrial membrane potential (MMP), mitochondrial superoxide (mtROS) generation, and intracellular calcium status were assessed at different time intervals during incubation by flow cytometry. It was observed that the dead sperm population increased (p < 0.01) during incubation in both the 20% and 30% SP-supplemented groups. However, no significant changes were observed in MMP in both the control and treatment groups at different time intervals. Interestingly, it was found that sperm mtROS production increased (p < 0.01) during incubation in the SP-supplemented groups compared with the control group. The proportion of live spermatozoa with high intracellular calcium was reduced (p < 0.01) during incubation in the SP-incubated groups. Collectively, heterologous SP addition could not repair the damages caused by the cryopreservation and further resulted in deterioration of semen quality as observed in our study by reducing viability, increasing reactive oxygen species (ROS) production possibly due to high proportion of dead cells, or some factors (yet to be identified) that are inducive of oxidative stress in stallion spermatozoa.


Potential energy surface and unimolecular dynamics of stretched n-butane.
The potential energy surface (PES) and unimolecular reaction dynamics of stretched n-butane are investigated, as a model for a stretched "normal" alkane or straight chain polymer. The nature of the PES for stretched n-butane depends on the extent of stretching. If it is less than that required to reach the inflection points in the C[Single Bond]C stretch potentials and the C[Single Bond]C torsions are considered free rotors, there is only one potential energy minimum, with each bond elongated. However, for stretching past these inflection points, the PES has three minima and each has one bond longer than the other two, i.e., C[Single Bond]C[Single Bond]C[Single Bond]C, C[Single Bond]C[Single Bond]C[Single Bond]C, and C[Single Bond]C[Single Bond]C[Single Bond]C. There are three transition states (TSs) connecting these minima. A linear alkane, consisting of n carbon atoms and stretched past its C[Single Bond]C inflection points, has (n-1) minima and (n-1)(n-2)2 TSs connecting them. For stretching less than that required to reach the C[Single Bond]C inflection points, the only unimolecular pathways are dissociations to form the C+C[Single Bond]C[Single Bond]C, C[Single Bond]C+C[Single Bond]C, and C[Single Bond]C[Single Bond]C+C products. However, with stretching past the C[Single Bond]C inflection points, isomerizations between the three potential energy minima may also occur. The relative importance of isomerization versus dissociation depends on the relative size of their barriers. For slight stretching past the C[Single Bond]C inflection points, the isomerization barriers are much lower than those for dissociation and relaxation between the minima is much faster than dissociation. Thus, the molecule samples these minima randomly during its dissociation, with a density of states that comprises the complete PES. With extensive stretching past the inflection points, isomerizations between the potential energy minima do not occur and only dissociation for the excited minima occurs, e.g., C[Single Bond]C[Single Bond]C[Single Bond]C-->C+C[Single Bond]C[Single Bond]C. For intermediate stretching past the C[Single Bond]C inflection points, the rates for the isomerization and dissociation pathways are competitive and both must be considered in modeling the dissociation kinetics. Microcanonical chemical dynamics simulations are performed to study the unimolecular kinetics of n-butane in three stretched configurations: Stretched less than the C[Single Bond]C inflection point; stretched slightly beyond the C[Single Bond]C inflection point; and stretched significantly beyond the C[Single Bond]C inflection point. The resulting unimolecular dynamics were found to be in excellent agreement with RRKM theory. Frequency nu factors, determined by fitting the trajectory unimolecular rate constants to the classical harmonic RRKM rate constant expression, depend upon the extent of stretching and vary from 1.0 x 10(12)-8.4 x 10(16) s(-1). For a molecule with a large number of vibrational degrees of freedom and high excess energy, it is shown that the classical harmonic RRKM and classical harmonic transition state theory rate constants, k(E) and k(T), are equivalent.


Structure characterization of the non-crystalline complexes of copper salts with native cyclodextrins.
The characterization of non-crystalline complexes is particularly difficult when techniques like X-ray diffraction or NMR cannot be used. We propose a simple procedure to characterize the physicochemical properties of amorphous new coordination compounds between cyclodextrins (CD) and Cu(2+) salts, by means of the integration of the information provided by several techniques including elemental analysis, flame atomic absorption, TGA, UV-Vis diffuse reflectance, colorimetry, FT-IR and EPR. On the basis of these procedures, we suggest geometrical and structural approximations resulting in an octahedral or distorted octahedral geometry with diverse positions for the metallic centre. According to the EPR spectrum, only one of the complexes may have rhombic symmetry. We also analyzed enthalpy-entropy compensation and the isokinetic effect. In addition, general trends in thermal stability, spectroscopic properties and inclusion in the cavity were analysed. This complete characterization methodology becomes essential for their future application as catalysts.


Nanostructured Brownian surfaces prepared through two-photon polymerization: investigation of stem cell response.
Nondeterministic phenomena are at the base of plenty of biological processes that comprise physiological signaling, cellular communications, and biological architectures. Among them, natural surface topographies are often characterized by "chaotic" features that are not trivial to be recreated in vitro. Recently, some methods have been proposed to resemble the hierarchical organization of the extracellular microenvironment, through the chemical preparation of randomly rough and self-affine fractal surfaces. Notwithstanding, this approach does not allow the fractal dimension to be modulated at a desired value, being moreover the self-affinity maintained just for one decade of spatial frequencies. Here, we propose the replication of in silico generated Brownian surfaces through a two-photon polymerization technique. As a result of the direct laser writing of the desired patterns, we were able to obtain highly reproducible self-affine (in a range of two spatial frequency decades) structures characterized by the desired predetermined Hurst exponents. Rat mesenchymal stem cells were moreover cultured on the obtained substrates, highlighting interesting phenomena concerning cell adhesion, cytoskeleton conformation, and actin polymerization, strictly depending on the fractal dimension of the surfaces.


Generation of even and odd high harmonics in resonant metasurfaces using single and multiple ultra-intense laser pulses.
High harmonic generation (HHG) opens a window on the fundamental science of strong-field light-mater interaction and serves as a key building block for attosecond optics and metrology. Resonantly enhanced HHG from hot spots in nanostructures is an attractive route to overcoming the well-known limitations of gases and bulk solids. Here, we demonstrate a nanoscale platform for highly efficient HHG driven by intense mid-infrared laser pulses: an ultra-thin resonant gallium phosphide (GaP) metasurface. The wide bandgap and the lack of inversion symmetry of the GaP crystal enable the generation of even and odd harmonics covering a wide range of photon energies between 1.3 and 3 eV with minimal reabsorption. The resonantly enhanced conversion efficiency facilitates single-shot measurements that avoid material damage and pave the way to study the controllable transition between perturbative and non-perturbative regimes of light-matter interactions at the nanoscale.


Metabolic activation of N-acylanthracyclines precedes their interaction with DNA topoisomerase II.
The N-acylanthracyclines AD32 (N-trifluoroacetyladriamycin-14-valerate) and AD143 (N-trifluoroacetyladriamycin-14-O-hemiadipate) are analogs of Adriamycin (ADR) undergoing clinical or advanced pre-clinical screening. Their principal metabolites, following the cleavage of the 14-acyl side-chain, are N-trifluoroacetyladriamycin (AD41) and its reduced form N-trifluoroacetyladriamycinol (AD92). Both these compounds are biologically active and detectable in treated patients, laboratory animals, and in tissue culture cells. Unlike ADR, AD32, as well as AD143 and metabolites, show no detectable binding to double-strand DNA. Their effects on DNA have been previously investigated in vivo and in vitro using the alkaline filter-elution assay. It has been shown that all of the compounds cause approximately equivalent amounts of protein-associated DNA breaks (PAB) and DNA-protein crosslinks in a mouse lymphoma and in tissue-culture leukemia cells. In order to establish whether the induction of PAB by the drugs requires DNA topoisomerase II mediation, cleavage mapping analysis was done with tested compounds using purified human topoisomerase II. DNA fragmentation was significantly enhanced in the presence of the enzyme and either AD41 or AD92. In contrast, no fragmentation enhancement was observed in the presence of the parental drugs AD32 or AD143. The results strongly suggest that metabolic activation of N-acylanthracyclines by nonspecific esterases is a prerequisite for their interaction with DNA topoisomerase II and for stabilization of the cleavable complex.


Nonmedical management of late luteal phase dysphoric disorder : a preliminary report.
The authors report the results of an open trial of group behavioral treatment for women with a prospectively confirmed premenstrual syndrome. Treatment consisted of five weekly group behavioral sessions outlining and supporting lifestyle changes in diet, exercise level, and stress management. Patients monitored symptoms and adherence to dietary and exercise recommendations daily. Results are discussed for the first 48 women to complete the trial. Eighty-three percent of patients demonstrated either a remission (n= 28) or 50% reduction in symptoms (n = 12) at one month following the group treatment. Improvements in physical and emotional symptoms were significantly associated with an increase in exercise.


Expression of transgenes targeted to the Gt(ROSA)26Sor locus is orientation dependent.
Targeting transgenes to a chosen location in the genome has a number of advantages. A single copy of the DNA construct can be inserted by targeting into regions of chromatin that allow the desired developmental and tissue-specific expression of the transgene. In order to develop a reliable system for reproducibly expressing transgenes it was decided to insert constructs at the Gt(ROSA)26Sor locus. A cytomegalovirus (CMV) promoter was used to drive expression of the Tetracycline (tet) transcriptional activator, rtTA2(s)-M2, and test the effectiveness of using the ROSA26 locus to allow transgene expression. The tet operator construct was inserted into one allele of ROSA26 and a tet responder construct controlling expression of EGFP was inserted into the other allele. Expression of the targeted transgenes was shown to be affected by both the presence of selectable marker cassettes and by the orientation of the transgenes with respect to the endogenous ROSA26 promoter. These results suggest that transcriptional interference from the endogenous gene promoter or from promoters in the selectable marker cassettes may be affecting transgene expression at the locus. Additionally we have been able to determine the optimal orientation for transgene expression at the ROSA26 locus.


Commentary on Current Trends in Rising Drug Costs and Reimbursement Below Cost.
To quantify prescription drug price increases over a span of 3 years (2012-2015), as well as extrapolate current reimbursement rates expected by independent retail pharmacies. In addition, we investigate potential reasons for these increasing drug costs. Descriptive analysis. National average drug acquisition costs (NADAC) data published by the Centers for Medicare & Medicaid Services were examined. Specifically, December 2012, 2013, and 2014, and July 2015 NADAC files were analyzed to identify generic and branded products with the highest percentage price increases. Percentage price differences were also calculated for 17 first-in-class drugs and their "me-too" competitors. The margin and margin percentage were calculated for claims adjudicated through four major payers. The top 50 generic drug price increases ranged from 474% to over 18,000% from December 2012 to July 2015. The top 50 branded drug price increases ranged from 63% to 391% during the same time period. The percentage price difference for the first-in-class drugs versus their me-too analogues ranged from -2.3% to 61,259%. The margin for generic drug claims adjudicated ranged from -$237.11 to-$1,105.96. The margin for branded drug claims adjudicated ranged from $272.42 to $360.17. Several potential reasons for the surge in prescription drug prices include manufacturer competition, industry consolidation, and capitalization on me-too drugs. This increase has compelled PBMs, health plan sponsors, and retail pharmacies to find novel ways to turn a profit, often at the expense of the consumer. Although there are no immediate solutions, legislation regulating PBM functions and the use of therapeutic interchange programs may offer health plans some assistance in managing drug costs.


[Fever : to treat or not to treat ?]
Fever is routinely faced during medical practice. Although it is part of the immune response against pathogens, it is often seen as deleterious and a source of discomfort leading to the use of antipyretic drugs. However, few clinical data support this common practice. On the contrary, to date, results tilt in favor of the beneficial effect of fever, such as a better immune response and inhibition of pathogen growth. Data encourage the treatment of fever only in certain conditions such as neurological symptoms, cardio-pulmonary failure, or fever of non-infectious origin. In fact recent evidence stress that systematic treatment of fever do not improve, but nor deteriorate, the disease outcome. This suggests to only treat the poorly tolerated fevers.


An anatomical subject-specific FE-model for hip fracture load prediction.
In order to reduce the socio-economic burden induced by osteoporotic hip fractures, finite element models have been evaluated as an additional diagnostic tool for fracture prediction. For a future clinical application, the challenge is to reach the best compromise between model relevance and computing time. Based on this consideration, the current study focused on the development and validation of a subject-specific FE-model using an original parameterised generic model and a specific personalization method. A total of 39 human femurs were tested to failure under a quasi-static compression in stance configuration. The corresponding FE- models were generated and for each specimen the numerical fracture load (FFEM) was compared with the experimental value (FEXP), resulting in a significant correlation (FEXP = 1.006 FFEM with r2 = 0.87 and SEE = 1220 N, p < 0.05) obtained with a reasonable computing time (30 mn). Further in vivo study should confirm the ability of this FE-model to improve the fracture risk prediction.


Decreased production and increased catabolism of apolipoprotein B-100 in apolipoprotein B-67/B-100 heterozygotes.
Apolipoprotein (apo) B-67 is a truncated form of apoB-100 due to deletion of an adenine at cDNA 9327. Heterozygotes have one allele making apoB-100; therefore, plasma apoB levels would be predicted to be at least 50% of normal. However, apoB-67 heterozygotes have total plasma apoB levels that are 24% of normal. To determine the mechanisms responsible for the lower-than-expected levels of apoB, in vivo kinetics of apoB-100 were performed in three apoB-67/apoB-100 heterozygotes and compared with those of six control subjects by using a primed-constant infusion of [5,5,5-2H3]leucine in the fed state. Kinetic parameters were calculated by multicompartmental modeling of the data. The mean total apoB plasma concentration of the apoB-67 subjects was 21.8+/-6.1 mg/dL, or 24% of that of control subjects (89.6+/-24.1 mg/dL, P=.002). ApoB-67 subjects had lower mean VLDL apoB-100 production rates (3.6+/-1.2 versus 13.9+/-3.5 mg x kg(-1) x d(-1), P=.002) and lower mean transport rates of apoB-100 into LDL (3.5+/-1.4 versus 12.6+/-4.1 mg x kg(-1) x d(-1), P=.008) compared with control subjects. The transport rate into IDL was not significantly different (1.2+/-0.5 versus 6.2+/-4.0 mg x kg(-1) x d(-1), P=.07). The fractional catabolic rate of VLDL apoB-100 was significantly higher in apoB-67 subjects than in control subjects (18.1+/-8.6 versus 7.6+/-1.6 mg x kg(-1) x d(-1), P=.017). ApoB-100 IDL and LDL fractional catabolic rates were not significantly different. VLDL apoB-100 pool size in apoB-67 subjects was 11% of that of control subjects (15.8+/-7.7 versus 141.6+/-33.7 mg, P=.0004) due to a 74% lower production rate (26% of control values) and a 2.4-fold higher fractional catabolic rate. LDL apoB-100 pool size in apoB-67 subjects was 22% of that of control subjects (665.3+/-192.4 versus 2968.3+/-765.2 mg, P=.002) due primarily to a lower production rate (27% of control values). Thus, both decreased production of VLDL and LDL apoB-100 and increased catabolism of VLDL apoB-100 are responsible for the low levels of apoB-100 in apoB-67 subjects.


Diversity of extended-spectrum beta-lactamase-producing Escherichia coli rods.
The aim of the study was to evaluate genetic relatedness and antimicrobial susceptibility of extended-spectrum beta-lactamase-producing E. coli strains isolated from patients hospitalized in the University Hospital in Bydgoszcz (Poland). The study included 33 extended-spectrum beta-lactamase-producing E. coli strains isolated from 31 patients. The chromosomal DNA was extracted from the strains and separated by pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was performed by the agar dilution method and carried out according to the European Committee on Antimicrobial Susceptibility Testing recommendations. According to the pulsed-field gel electrophoresis results, 32 distinct pulsotypes were revealed. Based on Molecular Analyst Fingerprinting software analysis, the studied isolates were divided into four subgroups: 6 (18.2%) isolates showing similarity greater than 90% (group A); 19 (57.6%) showing 80-90% similarity (group B); 7 (21.2%) showing 70-79% similarity (group C); and one isolate with less than 70% similarity (group D). Among E. coli isolates showing similarity greater than 90%, four antimicrobial patterns were noted. Among the isolates showing 80-90% similarity, 18 antimicrobial patterns were observed. E. coli isolates showing 70-79% similarity presented 6 antimicrobial patterns. Our results show a high degree of genetic diversity of extended-spectrum beta-lactamase-producing E. coli isolates. However, based on a similarity of ≥80%, almost 75% of E. coli isolates were clonally related. Although it is difficult to identify definitive transmission events based on the recovery of indistinguishable pulsed-field gel electrophoresis types alone, we speculate that extended-spectrum beta-lactamase-producing E. coli strains may have disseminated throughout the hospital.


Metal Complexes of a Novel Schiff Base Based on Penicillin: Characterization, Molecular Modeling, and Antibacterial Activity Study.
A novel Schiff base ligand of type HL was prepared by the condensation of amoxicillin trihydrate and nicotinaldehyde. The metal complexes of Co+2, Ni+2, Cu+2, and Zn+2 were characterized and investigated by physical and spectral techniques, namely, elemental analysis, melting point, conductivity, 1H NMR, IR, UV-Vis spectra, ESR, SEM, and mass spectrometry measurements. They were further analyzed by thermal technique (TGA/DTA) to gain better insight about the thermal stability and kinetic properties of the complexes. Thermal data revealed high thermal stability and nonspontaneous nature of the decomposition steps. The Coats-Redfern method was applied to extract thermodynamic parameters to explain the kinetic behavior. The molar conductance values were relatively low, showing their nonelectrolytic nature. The powder XRD pattern revealed amorphous nature except copper complex (1c) that crystallized in the triclinic crystal system. The EPR study strongly recommends the tetrahedral geometry of 1c. The structure optimization by MM force field calculation through ArgusLab 4.0.1 software program supports the concerned geometry of the complexes. The in vitro antibacterial activity of all the compounds, at their two different concentrations, was screened against four bacterial pathogens, namely, E. coli, P. vulgaris, K. pneumoniae, and S. aureus, and showed better activity compared to parent drug and control drug.


Selection of Metastatic Breast Cancer Cell-Specific Aptamers for the Capture of CTCs with a Metastatic Phenotype by Cell-SELEX.
Circulating tumor cells (CTCs) have the potential to predict metastasis, and the capture of CTCs based on their surface markers is mostly applied for CTC detection. Considering that the CTCs with a metastatic phenotype preferably form a metastatic focus and that aptamers have the ability to bind targets with high specificity and affinity, we selected aptamers directed toward metastatic cells by subtractive Cell-SELEX technology using highly metastatic MDA-MB-231 cells as the target cell and low-metastatic MCF-7 cells as the negative cell for the capture of metastatic CTCs. Affinity and selectivity assays showed that aptamer M3 had the highest affinity, with a KD of 45.6 ± 1.2 nM, and had good specificity against several other types of metastatic cancer cells. Based on these findings, we developed an M3-based capture system for CTC enrichment, which has the capability to specifically capture the metastatic cells MDA-MB-231 mixed with non-metastatic MCF-7 cells and CTCs derived from the peripheral blood from metastatic breast cancer patients. A further comparative analysis with the anti-EpCAM probe showed that M3 probe captured epithelial feature-deletion metastatic cells. We developed an aptamer-based CTC capture system through the selection of aptamers by taking whole metastatic cells, not known molecules, as targets, which provided a new insight into CTC capture and Cell-SELEX application.


K-site splitting in KTiOPO4 at room temperature.
The room-temperature structure of potassium titanyl phosphate (KTiOPO4, KTP) with Pna2(1) symmetry has been studied by means of synchrotron radiation. Each of the two crystallographically unique K1 and K2 cations is split over two sites that are shifted along the c direction by 0.287 (13) and 0.255 (13) A for the K1a/b and K2a/b pairs, respectively. The refined populations of the minor K1b and K2b sites are 0.102 (12) and 0.132 (17), respectively. It is shown that accurate high-resolution synchrotron data (Rmerged = 0.015 for 25 010 reflections, 9456 unique, sintheta/lambda limit > 1.0) are required for the determination of a reliable structure model.


Dried blood spot sampling: coupling bioanalytical feasibility, blood-plasma partitioning and transferability to in vivo preclinical studies.
The adoption of dried blood spot (DBS) sampling and analysis to support drug discovery and development requires the understanding of its bioanalytical feasibility as well as the distribution of the analyte in blood. Demonstrated the feasibility of adopting DBS for four test analytes representing diverse physico-chemical as well as pharmacokinetic parameters. The key findings include the use of a single extraction procedure across all four analytes, assay range of 1 to 5000 ng/ml, stability in whole blood as well as on-card, and the non-impact of blood volume. In vivo data were used to calculate the blood-to-plasma ratio (using both AUC and average of individual time points), which was then used to predict plasma concentration from DBS data. The predicted data showed an excellent correlation with actual plasma data. Transition from plasma to DBS can be supported for preclinical studies by conducting a few well-defined bioanalytical experiments followed by an in vivo bridging study. Blood:plasma ratio derived from the bridging study can be used to predict plasma concentrations from DBS data.


Normal and Injured Ankle Ligaments on Ultrasonography With Magnetic Resonance Imaging Correlation.
Ultrasonography (US) has been increasingly used in the evaluation of ankle ligamentous injuries given its advantages as a dynamic, efficient, noninvasive, and cost-effective imaging method. Understanding the anatomy of the ankle ligaments is critical for correct diagnosis and treatment. This pictorial essay describes and illustrates the US scanning technique and potential pitfalls in evaluating the ankle ligaments and also provides an overview of the US appearance of normal and injured ankle ligaments with magnetic resonance imaging correlation. Highlighted structures include the lateral complex, medial/deltoid complex, spring (calcaneonavicular) ligament complex, and syndesmosis.


Alteration in the relationship between tanycytes and gonadotrophin-releasing hormone neurosecretory terminals following long-term metabolic manipulation in the sheep.
The activity of the hypothalamic-pituitary gonadal axis is influenced by energy reserves, such that an increase or a decrease in adiposity may perturb the secretion and action of gonadotrophin-releasing hormone (GnRH). This is considered to be a result of the signalling of hormones such as leptin, which act upon neuronal systems controlling GnRH secretion. Other work shows plasticity in the relationship between tanycytes and GnRH neurosecretory terminals in the median eminence across the oestrous cycle and we hypothesised that a similar plasticity may occur with altered metabolic status. We studied Lean, Normal and Fat ovariectomised ewes, which displayed differences in gonadotrophin status, and investigated the relationship between tanycytes and GnRH neuroterminals. Under both Lean and Fat conditions, an altered anatomical arrangement between these two elements was observed in the vicinity of the blood vessels of the primary plexus of the hypophysial portal blood system. These data suggest that such plasticity is an important determinant of the rate of secretion of GnRH in animals of differing metabolic status and that this also contributes to the relative hypogonadotrophic condition prevailing with metabolic extremes.


Mechanism of ACh-induced asynchronous calcium waves and tonic contraction in porcine tracheal muscle bundle.
Stimulation of the tracheal muscle bundle by acetylcholine (ACh) results in the generation of asynchronous repetitive Ca2+ waves (ACW) in intact tracheal smooth muscle (TSM) cells. We showed previously that ACW underlie cholinergic excitation-contraction coupling in porcine TSM and that Ca2+ entry through the L-type voltage-gated Ca2+ channel (VGCC) contributes partially to maintenance of the ACW. However, the mechanism of the ACW remains undefined. In this study, we pharmacologically characterized the mechanism of ACh-induced ACW in the intact porcine tracheal muscle bundle. We found that inhibition of receptor-operated channels/store-operated channels (ROC/SOC) by SKF-96365 completely abolished the nifedipine-insensitive component of ACh-mediated ACW and tonic contraction. Blockade of Na+/Ca2+ exchange with KB-R7943 or 2',4'-dichlorobenzamil or removal of extracellular Na+ resulted in nearly complete inhibition of the nifedipine-insensitive component of ACh-mediated ACW and tonic contraction. Inhibition of the sarco(endo)plasmic reticulum Ca2+-ATPase by cyclopiazonic acid abolished the ongoing ACW. Application of 2-aminoethoxydiphenyl borate (2-APB) or xestospongin C to inhibit the inositol 1,4,5-trisphosphate-sensitive sarcoplasmic reticulum (SR) Ca2+ release channels produced no effect on ACh-mediated ACW and tonic contraction. However, pretreatment with caffeine or ryanodine inhibited ACh-induced ACW. Furthermore, application of procaine or tetracaine prevented the generation and abolished the ongoing ACh-mediated ACW and tonic contraction. Collectively, these results indicate that the ACh-stimulated ACW in porcine TSM are produced by repetitive cycles of Ca2+ release from SR through 2-APB- and xestospongin C-insensitive Ca2+ release channels, and plasmalemmal Ca2+ entry involving reverse-mode Na+/Ca2+ exchange, ROC/SOC, and L-type VGCC is required to refill the SR via SERCA to support the ongoing ACW.


CYP2D6 Pharmacogenetics Testing and Post-Cesarean Section Pain Scores-a Preliminary Study.
Prospective observational study to analyze CYP2D6 pharmacogenetics in 55 Portuguese adult parturients undergoing elective cesarean section and to investigate the association between CYP2D6 alleles and pain score. DNA was extracted from peripheral blood by standard methods. Genetic analysis included allelic discrimination (CYP2D6*1, *2, *3, *4, *5, *6, *10, *17, and *41) and copy number determination with TaqMan probes by real-time polymerase chain reaction (PCR). Allele duplications were confirmed (long PCR and PCR-restriction fragment length polymorphism). Theoretical metabolic profiles prediction was based on genetic data and activity scores. Association was investigated between genotypes and predicted phenotypes with pain scores. Statistical analysis was performed by using a χ2 test, and significance was set at P < 0.05. The percentage of poor, intermediate, extensive, and ultrarapid metabolizers found were 9%, 38%, 46%, and 7%, respectively. The results reveal a positive association between alleles *4, *10, and pain. A positive association was found between predicted reduced or null activity of CYP2D6 and increased pain. It can be hypothesized that if CYP2D6 activity is reduced, tyramine metabolism will decrease, resulting in reduced formation of endogenous dopamine. Consequently, activation of the signal transduction pathways that controls pain and analgesic effect may be reduced, leading to an increase in pain. Therefore, we would recommend CYP2D6 genotyping to anticipate the needs for analgesia, which will help to adjust opioid dose and maximize clinical efficacy while reducing side effects.


Retroperitoneal bronchogenic cyst: a case report.
Bronchogenic cysts are among developmental disorders of the primitive foregut which are typically found above the diaphragm. Bronchial cysts discovered in the abdominal cavity or retroperitoneum are extremely rare. We present a rare case of a retroperitoneal bronchogenic cyst which was incidentally detected after a wrestling injury in a 23-year-old man who had a negative medical history. Although initial imaging studies suggested an adrenal tumor, histopathological analysis provided a definite diagnosis of bronchogenic cyst. Though rare, bronchogenic cysts must be considered in the differential diagnosis of retroperitoneal cystic lesions. This is the first case of a retroperitoneal bronchogenic cyst reported in Iran.


DNA mismatch repair detected in human cell extracts.
A system to study mismatch repair in vitro in HeLa cell extracts was developed. Preformed heteroduplex plasmid DNA containing two single base pair mismatches within the SupF gene of Escherichia coli was used as a substrate in a mismatch repair assay. Repair of one or both of the mismatches to the wild-type sequence was measured by transformation of a lac(Am) E. coli strain in which the presence of an active supF gene could be scored. The E. coli strain used was constructed to carry mutations in genes associated with mismatch repair and recombination (mutH, mutU, and recA) so that the processing of the heteroduplex DNA by the bacterium was minimal. Extract reactions were carried out by the incubation of the heteroduplex plasmid DNA in the HeLa cell extracts to which ATP, creatine phosphate, creatine kinase, deoxynucleotides, and a magnesium-containing buffer were added. Under these conditions about 1% of the mismatches were repaired. In the absence of added energy sources or deoxynucleotides, the activity in the extracts was significantly reduced. The addition of either aphidicolin or dideoxynucleotides reduced the mismatch repair activity, but only aphidicolin was effective in blocking DNA polymerization in the extracts. It is concluded that mismatch repair in these extracts is an energy-requiring process that is dependent on an adequate deoxynucleotide concentration. The results also indicate that the process is associated with some type of DNA polymerization, but the different effects of aphidicolin and dideoxynucleotides suggest that the mismatch repair activity in the extracts cannot simply be accounted for by random nick-translation activity alone.


Androgen receptor (AR) expression in normal and cancerous human endometrial tissues detected by RT-PCR and immunohistochemistry.
This study aimed at assessing the expression and cellular localisation of the androgen receptor (AR) in normal and malignant human endometrial tissues. Twelve normal endometrial samples and 25 uterine cancers were studied using RT-PCR and immunohistochemistry (IHC). One of the 12 (8%) normal endometrial samples showed AR PCR-product, whereas 4 of the 25 (16%) cancers displayed AR expression. Tumor tissues and normal tissues did not differ significantly in AR expression (p=0.66; Mann-Whitney U-test). Nuclear staining for AR was positive in 10 of the 25 (40%) cancers, in the glandular cancer cells, in the surrounding stroma and in the myometrium. There was a significant correlation between mRNA RT-PCR AR expression and the IHC AR staining in endometrial tumors (r=0.53; p=0.006). To the best of our knowledge, this is the first description of AR expression in both normal and pathological changes of the human endometrium using the RT-PCR techniques. Our observations confirmed the role of the AR in the stromal regulation of epithelial cells within the neoplastic endometrium.


Empiric therapy for cancer patients: comparative study of ticarcillin-tobramycin, ticarcillin-cephalothin, and cephalothin-tobramycin.
Three combinations of antibiotics (cephalothin-tobramycin, cephalothin-ticarcillin, and ticarcillin-tobramycin) were administered empirically to 186 patients with cancer who were suspected of having a life-threatening infection. In approximately one-half of these patients, gram-negative infection was documented bacteriologically and consisted of septicemia in 50% of these patients. The three antimicrobial regimens were similarly effective and resulted in a favorable clinical response in approximately 55% of the patients. The administration of the cephalothin-tobramycin combination was associated with a significantly higher frequency of nephrotoxicity than that of the other two regimens.


Principles and theory in bioethics.
The following citations were selected from BIOETHICSLINE, the online database prepared at the Kennedy Institute of Ethics for the National Library of Medicine's MEDLARS system. Searching the keywords autonomy, beneficence, casuistry, justice, and virtues, as well as the text word principlism produced more than 400 citations. Only the citations concerned with theory and principle in the practice of bioethics are included here -- e.g., works about justice in resource allocation have been deleted.


Increased Synovial Fluid YKL-40 Levels are Linked with Symptomatic Severity in Knee Osteoarthritis Patients.
Elevated serum and synovial fluid (SF) YKL-40 levels have been detected in knee osteoarthritis (OA) patients. The current study was focused on the correlation between YKL-40 levels in serum or SF and symptomatic severity in patients with knee osteoarthritis. 144 patients with knee OA and 151 healthy individuals were recruited into this study. Symptomatic severity was determined using Western Ontario and McMaster Universities Arthritis Index (WOMAC) scores from OA patients. Serum and SF levels of YKL-40 were explored by enzyme-linked immunosorbent assay. We found that YKL-40 levels in SF but not serum were independently and positively related to WOMAC pain (r = 0.531, p = 0.001), physical disability (r = 0.380, p = 0.025), and total scores (r = 0.407, p = 0.01) in knee OA patients. YKL-40 in SF could represent a potential biomarker for assessing the symptomatic severity of OA.


Synchronous adenocarcinoma and lymphoma of the colon.
Synchronous lymphoma and adenocarcinoma of the colon are extremely rare. A 32-year-old woman was referred to us for colon cancer. Investigations revealed two primary tumors, one in the cecum and the other in the sigmoid colon. Subtotal colectomy with ileorectal anastomosis was performed. Histology revealed the cecal tumor to be non-Hodgkin's lymphoma, diffuse small cell type with plasmacytoid features. The sigmoid colon tumor was moderately differentiated adenocarcinoma. The patient received 6 cycles of chemotherapy postoperatively for lymphoma but died of recurrent disease after 17 months.


[The mental health paradigmatic transition].
This work is a result of authors' reflection on the necessity of conceptual changes in the area of mental health. The authors aimed at understanding the essential aspects involving the theme according to the conception of paradigm, as well as the process in which the crisis appears emerging anomalies and pointing out the inefficiency of the present paradigm. Thus, they suggested a change from the actual model to another one based on a perspective that emphasizes patient's freedom and the relational character of human expressiveness.


Comparison of HMOX1 expression and enzyme activity in blue-shelled chickens and brown-shelled chickens.
Blue egg coloring is attributed to biliverdin derived from the oxidative degradation of heme through catalysis by heme oxygenase (HO). The pigment is secreted into the eggshell by the shell gland. There is uncertainty as to whether the pigment is synthesized in the shell gland or in other tissues. To investigate the site of pigment biosynthesis, the expression of heme oxygenase (decycling) 1 (HMOX1), a gene encoding HO, and HO activity in liver and spleen were compared between blue-shelled chickens (n = 12) and brown-shelled chickens (n = 12). There were no significant differences in HMOX1 expression and HO activity in these tissues between the two groups. Since the liver and spleen, two important sites outside the shell gland where heme is degraded into biliverdin, CO and Fe(2+), did not differ in HO expression and activity we conclude that the pigment is most likely synthesized in the shell gland.


The cross-sectional survey on COVID-19 vaccine hesitancy and it predictors among Chinese parents of 3-17 years aged children in Shenzhen City.
Vaccinations programs on 3-17 years aged children in China have been launched in some cities since July 2021; and comparative evaluations are important to push the programs forward. Therefore, this study is conducted to explore the COVID-19 vaccine hesitancy and it predictors among Chinese parents of 3-17 years aged children; and their willingness to vaccinate their child/children. A cross-sectional study was conducted based on the online survey; and 3484 participants were recruited in health centers of Shenzhen, China. The prevalence of COVID-19 vaccine hesitancy was 20.7% among Chinese parents of 3-17 years aged children; and 2976 participants (89.0%) were reported to be willing or very willing to accept a COVID-19 vaccine for their child/children. Perceived trust degree of a COVID-19 vaccine (OR: 0.766), and perceived effectiveness of a COVID-19 vaccine (OR: 0.455) were negatively associated with vaccine hesitancy. Perceived risk degree of a COVID-19 vaccine (OR: 1.485) positively predicted vaccine hesitancy. Parents with a higher educational level (OR: 0.518) and working in public institution (OR: 0.741) had a lower level of vaccine hesitancy; whereas, parents with liberal professionals had a higher level of vaccine hesitancy (OR: 1.378). These findings indicated that Chinese parents of 3-17 years aged children in Shenzhen City had a higher level of COVID-19 vaccine acceptability, compared with adults in Western counties. The significant factors of perceived trust degree, effectiveness, and risk degree of a COVID-19 vaccine, parents' educational level and occupation could predict COVID-19 vaccine hesitancy. The research results can be useful in supporting the development of effective and targeted children COVID-19 vaccination campaigns in China and worldwide.


Alexithymia and body weight in obese patients.
The authors evaluated severely obese patients to determine whether being far different in body shape from the accepted standard may cause obese people to develop alexithymic personality traits. They evaluated the food- and weight-related attitudes in obesity surgery patients and in long-term follow-up of those who had previously had biliopancreatic diversion (BPD) for obesity. One quarter of the obese patients had alexithymic characteristics without any modification following stable weight loss, a rate of alexithymia similar to that observed in the nonclinical population. Furthermore, the frequency of alexithymia and the patients' scores on the Toronto Alexithymia Scale were similar in obese and post-BPD individuals. The authors concluded that being obese by itself does not influence the presence of alexithymic personality traits. However, they suggest that the improvement in food-related and weight-related attitudes following stable weight loss may be different in alexithymic and in nonalexithymic obese patients.


LYTIC CAPABILITY OF BACTERIOPHAGES (FAMILY MYOVIRIDAE) ON BURKHOLDERIA PSEUDOMALLEI.
Burkholderia pseudomallei, a gram-negative bacillus found in soil and water, is the causative agent of melioidosis. It can produce a biofilm, which increases resistance to antibacterial agents. Bacteriophages (phages) have been suggested as alternative antibacterial agents. In this study, the ability of six phages (family Myoviridae) to lyse B. pseudomallei isolates was examined using a microplate phage virulence assay. The six phages were more efficient in lysing soil than clinical B. pseudomallei isolates. Phage ST79 had the highest lytic capability, independent of inoculating phage quantity with a 4-log reduction of bacterial numbers after a 4 hour treatment. Three modified derivatives of ST79 were developed by multiple passages on phage-resistant B. pseudomallei isolates, leading to an increase in lytic capability from 62% to 80%. Phage ST79 at a multiplicity of infection (MOI) of 10 significantly reduces biofilm formation determined by a colorimetric method. The recovery of B. pseudomallei growth following phage treatment needs to be overcome if these lytic phages are to be used as biocontrol agents of B. pseudomallei in the environment.


Characterization and distribution of polycyclic aromatic hydrocarbon in sediments of Haihe River, Tianjin, China.
In this study sediment samples were collected from 13 sites of Haihe River in Tianjin City, China, sixteen of priority polycyclic aromatic hydrocarbons (PAHs) listed in USEPA were analyzed by means of GC-MS. The total concentrations of PAH ranged from 774.81 to 255371.91 ng/g dw, and two to four rings of PAHs were dominant in sediment samples. Molecular ratios, such as phenanthrene/anthracene, fluoranthene/pyrene and low-molecular-weight PAH /high-molecular-weight PAH, were used to study the possible sources of pollution. It indicated a mixed pattern of parolytic and petrogenic inputs of PAHs in sediments in Haihe River. The petrogenic PAHs may be mainly derived from the leakage of refined products, e.g., gasoline, diesel fuel and fuel oil vehicle traffics or gas stations from urban area. The pyrolytic PAHs might be from the discharge of industrial wastewater and the emission of atmospheric particles from petrochemical factories. In addition, the levels of PAHs in the urban and industrial areas are far beyond the values reported from other rivers and marine systems reported. This situation may be due to polluted discharging from some petrochemical industrial manufactories and worse traffic conditions in Tianjin.


Synthesis and studies on antioxidants: ethoxyquin (EQ) and its derivatives.
In our study ethoxyquin (EQ) and its two complexes with flavonoids were obtained from ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline, EQ) and quercetin (EQ-Q, 1:1) or rutin (EQ-R, 1:1). Cytotoxicity of the tested compounds was studied using the trypan blue exclusion method and the properties of the studied compounds were also analyzed with the TUNEL method evaluating their ability to induce apoptosis. It was shown that EQ induced apoptosis in cultured human lymphocytes, especially at 0.25 and 0.5 mM concentrations. The same effects were also observed after the incubation of lymphocytes with EQ-Q and EQ-R, but the numbers of apoptotic cells observed were lower than for EQ.


Evaluation of the botanical origin of commercial dry bee pollen load batches using pollen analysis: a proposal for technical standardization.
High quality of bee pollen for commercial purpose is required. In order to attend the consumer with the best identification of the botanical and floral origin of the product, 25 bee pollen batches were investigated using two techniques of pollen grain preparation. The first started to identify pollen loads of different colors in two grams of each well mixed batch, and the second to identify pollen grains in a pool made of all the pollen loads comprised in two grams. The best result was obtained by this last technique, when a pollen grain suspension was dropped on a microscope slide and circa 500 pollen grains were counted per sample. This analysis resulted in the recognition of monofloral and bifloral pollen batches, while the use of the first technique resulted in all samples receiving a heterofloral diagnosis.


CONSIDERATIONS ON ANATOMY AND PHYSIOLOGY OF LYMPH VESSELS OF UPPER AERO DIGESTIVE ORGANS AND CERVICAL SATELLITE LYMPH NODE GROUP.
The almost constant local regional development of the cancers of upper aero digestive organs requires the same special attention to cervical lymph node metastases, as well as to the primary neoplastic burning point. The surgical therapy alone or associated has a mutilating, damaging character, resulting in loss of an organ and function, most of the times with social implications, involving physical distortions with aesthetic consequences, which make the reintegration of the individual into society questionable. The problem of cervical lymph node metastases is vast and complex, reason why we approached several anatomical and physiological aspects of lymph vessels of the aero digestive organs. Among the available elements during treatment, the headquarters of the tumour, its histologic degree, and its infiltrative nature, each of them significantly influences the possibility of developing metastases.


Assessment of soft- and hard-tissue changes following combined anterior segmental bi-jaw orthognathic surgery.
Anterior segmental Bi-jaw orthognathic surgery is indicated primarily for the correction of dentoalveolar protrusion. They are also indicated for correcting apertognathia, closing interproximal spaces between segments and can be incorporated with other osteotomies to obtain better results. The aim of this study was to analyze and compare the soft- and hard-tissue changes in patients who underwent combined anterior segmental bi-jaw orthognathic surgery. To compare and evaluate soft- and hard-tissue changes before and after combined anterior segmental bi-jaw orthognathic surgery by assessing (a) parameters in vertical dimension and (b) parameters in horizontal dimension and patient satisfaction was also assessed following surgery at 6 months' time interval. It is a prospective, single center and analytical study with sample size of 20 patients. It required routine setup for orthognathic surgery and manual tracing of lateral cephalograms. The surgical outcomes were assessed by hard tissue (Sella Nasion Point A (SNA) angle, Sella Nasion Point B (SNB) angle, Point A Nasion Point B (ANB) angle, NAPg, U1-NF, L1-MP angles; NA, NB, B-Pg, Nasion-Anterior nasal spine (N-ANS) distance, Anterior nasal spine-Gnathion (ANS-Gn) distance, and overjet and overbite) and soft tissue (facial convexity, NL, LM angles; LM fold, UL and LL protrusions, Upper lip length (ULL), Lower lip length (LLL), Sn-A, Si-B, Pg-Pg*, Ls-U1, Li-L1 distances, interlabial gap, and U1 exposure) variables pre and postsurgery. Descriptive statistics involved the mean and standard deviation, and recorded data were subjected to the statistical analysis using IBM SPSS 20 statistical package. The paired t-test, Pearson correlation coefficient were used. The level of significance P < 0.05 was taken as statistically significant and P < 0.01 as very significant. Overall improvement after surgery in both hard- and soft-tissue parameters was observed and significant changes were seen in every variable, except Si-B (P > 0.05). Correlations between soft- and hard-tissue changes were significant in both sagittal and vertical planes. Patients' satisfaction score showed that all patients had the high overall rate of satisfaction. Our study concludes that combined anterior segmental bimaxillary orthognathic surgery is a simple technique with minimal postoperative complications and limited relapse. The changes in facial esthetics and occlusion following orthognathic surgery depend highly on the stability achieved during the postoperative period.


Topical hyperglycemia rapidly suppresses EDRF-mediated vasodilation of normal rat arterioles.
Arteriolar dilation to endothelium-derived relaxing factor (EDRF) is suppressed early in diabetes mellitus. The purpose of this study was to determine whether acute exposure to a hyperglycemic media can suppress EDRF function of normal arterioles. Dilation of intestinal arterioles to iontophoretically applied acetylcholine (ACh) and nitroprusside was measured in normoglycemic rats before and after 1 h of topical exposure to isotonic solutions containing D-glucose concentrations of 200, 300, and 500 mg/100 ml. Exposure to a D-glucose concentration of 200 mg/100 ml had no effect on vasodilation to ACh. D-Glucose concentrations of both 300 and 500 mg/100 ml caused significant suppression of the responses: for example, at the approximate 50% effective dosage (100 nA), the dilatory response was decreased by 60% at a D-glucose concentration of 300 mg/100 ml and 55% at a D-glucose concentration of 500 mg/100 ml. Responses to nitroprusside were not significantly (P < 0.05) impaired after exposure to D-glucose concentrations of 200, 300, or 500 mg/100 ml. Exposure to an isotonic L-glucose concentration of 500 mg/100 ml for 1 h had no significant (P > 0.05) effect on responses to ACh. Pretreatment with superoxide dismutase, catalase, indomethacin, or meclofenamic acid preserved EDRF-mediated vasodilation during exposure to a D-glucose concentration of 500 mg/100 ml at almost all the ACh dosages tested. These results indicate that oxygen radicals formed in part by increased eicosanoid synthesis during exposure to D-glucose hyperglycemia interfere with the EDRF mechanism before its action on the microvascular smooth muscle.


Novel Dielectric Coagulometer Identifies Hypercoagulability in Patients with a High CHADS2 Score without Atrial Fibrillation.
Recent reports showed that the CHADS2 score predicted the risk of strokes in patients without atrial fibrillation (AF). Although the hypercoagulability may contribute to the thrombogenesis, it has not been fully investigated due to a lack of a sensitive evaluation modality. Recently a novel dielectric blood coagulometry (DBCM) was invented for evaluating the coagulability by measuring the temporal change in whole blood dielectric permittivity. We evaluated the utility of the DBCM for identifying the coagulability. For fundamental experiments, 133 citrated blood samples were drawn from subjects with or without heparin administration. A DBCM analysis was performed to find the adequate coagulation index, and to delineate its measurement range by adding recombinant human tissue factor (TF) or heparin. Then the coagulability was assessed by DBCM and conventional coagulation assays in 84 subjects without AF, who were divided into 3 groups by their CHADS2 score. Another 17 patients who received warfarin were also assessed by DBCM to evaluate the effect of anticoagulants. We calculated the derivative of the dielectric permittivity change after recalcification, and extracted the end of acceleration time (EAT) as a novel index. The EAT showed a dose-dependent shortening with the addition of serial dilutions of TF (×10-2 to ×10-4), and a dose-dependent prolongation with the addition of heparin (0.05 to 0.15 U/ml). The EAT was significantly shorter in the higher CHADS2 score group (19.8 ± 4.8, 18.6 ± 3.1, and 16.3 ± 2.7 min in the CHADS2 = 0, 1, and ≥2 groups, respectively, p = 0.0065 by ANOVA). Patients receiving warfarin had a significantly more prolonged EAT than those without warfarin (18.6±4.2 vs. 25.8±7.3 min, p <0.001). DBCM detected the whole blood coagulability with a high sensitivity. Subjects with higher CHADS2 scores exhibited hypercoagulability without AF.


Ifenprodil and ethanol enhance NMDA receptor-dependent long-term depression.
Long-term alterations in synaptic transmission are thought to underlie various types of alcohol-related brain disorders. While ethanol effects on synaptic potentiation are well documented, ethanol effects on synaptic depression have not been addressed. Herein, we performed experiments to assess the role of ethanol on long-term depression (LTD) formation. In rat hippocampal slices, prolonged low-frequency stimulation (LFS) of CA1 Schaffer collaterals (1 Hz for 7 min) induced saturable, long-lasting, reversible N-methyl-D-aspartate (NMDA) receptor-dependent LTD of stimulus-evoked dendritic population excitatory postsynaptic potentials. This depression (-26% LTD amplitude) was observed in young rats (12-20 days old), but not adult rats (28-35 days old). Induction of LTD was blocked (-3% LTD amplitude) when the LFS was delivered in the presence of the NMDA receptor antagonist D-2-amino-5-phosphonovaleric acid. When the conditioning LFS was delivered in the presence of ethanol, there was a significant enhancement in the induction of NMDA receptor-dependent LTD versus control LTD (-36% LTD amplitude). Ifenprodil, an N-methyl-D-aspartate receptor subunit 2B (NR2B)-selective antagonist, also significantly facilitated the induction of LTD (-40% LTD amplitude). Consistent with this result, ifenprodil did not affect the NMDA receptor-dependent component of the baseline synaptic response, whereas D-2-amino-5-phosphonovaleric acid caused significant depression of the NMDA component. These data indicate that whereas ethanol is known to inhibit NMDA receptor function in a variety of systems, it significantly enhances the induction of NMDA receptor-dependent LTD. Furthermore, since ifenprodil is known to select for ethanol-sensitive subtypes of NR2B-NMDA receptors, these data also suggest that NR2B-containing NMDA receptor subpopulations do not contribute to LTD, but instead may actually play inhibitory roles in LTD induction.


Long-distance signaling in bypass1 mutants: bioassay development reveals the bps signal to be a metabolite.
Root-to-shoot signaling is used by plants to coordinate shoot development with the conditions experienced by the roots. A mobile and biologically active compound, the bps signal, is over-produced in roots of an Arabidopsis thaliana mutant called bypass1 (bps1), and might also be a normally produced signaling molecule in wild-type plants. Our goal is to identify the bps signal chemically, which will then allow us to assess its production in normal plants. To identify any signaling molecule, a bioassay is required, and here we describe the development of a robust, simple, and quantitative bioassay for the bps signal. The developed bioassay follows the growth-reducing activity of the bps signal using the pCYCB1;1::GUS cell cycle marker. Wild-type plants carrying this marker, and provided the bps signal through either grafts or metabolite extracts, showed reduced cell division. By contrast, control grafts and treatment with control extracts showed no change in pCYCB1;1::GUS expression. To determine the chemical nature of the bps signal, extracts were treated with RNase A, Proteinase K, or heat. None of these treatments diminished the activity of bps1 extracts, suggesting that the active molecule might be a metabolite. This bioassay will be useful for future biochemical fractionation and analysis directed toward bps signal identification.


PYRIDOX(AM)INE 5'-PHOSPHATE OXIDASE3 of Arabidopsis thaliana maintains carbon/nitrogen balance in distinct environmental conditions.
The identification of factors that regulate C/N utilization in plants can make a substantial contribution to optimization of plant health. Here, we explored the contribution of pyridox(am)ine 5'-phosphate oxidase3 (PDX3), which regulates vitamin B6 homeostasis, in Arabidopsis (Arabidopsis thaliana). Firstly, N fertilization regimes showed that ammonium application rescues the leaf morphological phenotype of pdx3 mutant lines but masks the metabolite perturbance resulting from impairment in utilizing soil nitrate as a source of N. Without fertilization, pdx3 lines suffered a C/N imbalance and accumulated nitrogenous compounds. Surprisingly, exploration of photorespiration as a source of endogenous N driving this metabolic imbalance, by incubation under high CO2, further exacerbated the pdx3 growth phenotype. Interestingly, the amino acid serine, critical for growth and N management, alleviated the growth phenotype of pdx3 plants under high CO2, likely due to the requirement of pyridoxal 5'-phosphate for the phosphorylated pathway of serine biosynthesis under this condition. Triggering of thermomorphogenesis by growth of plants at 28 °C (instead of 22 °C) did not appear to require PDX3 function, and we observed that the consequent drive toward C metabolism counters the C/N imbalance in pdx3. Further, pdx3 lines suffered a salicylic acid-induced defense response, probing of which unraveled that it is a protective strategy mediated by nonexpressor of pathogenesis related1 (NPR1) and improves fitness. Overall, the study demonstrates the importance of vitamin B6 homeostasis as managed by the salvage pathway enzyme PDX3 to growth in diverse environments with varying nutrient availability and insight into how plants reprogram their metabolism under such conditions.