Interleukin-6 knockout reverses macrophage differentiation imbalance and alleviates cardiac dysfunction in aging mice.
Several interleukins (ILs) have been shown to be involved in aging, but the effects of IL-6 on aging-related cardiac dysfunction remain unknown. In this study, the expression and sources of cardiac IL-6 in aging hearts were investigated for the first time. The results showed that cardiac IL-6 expression in mice gradually increased with age, and the expression at 16 months, 20 months and 25 months was higher than that at 3 months. In addition, cardiac macrophages (Møs) were shown to be the main sources of IL-6 in aging mice. IL-6 knockout (KO) significantly alleviated cardiac dysfunction, increased M2 macrophage (Mø2) differentiation, reduced M1 macrophage (Mø1) differentiation and protected against cardiomyocyte apoptosis in aging mice. IL-6 KO also reversed the stimulatory effect of doxorubicin (DOX) treatment on Mø1s and the inhibitory effect of DOX treatment on Mø2s in vitro. Furthermore, the mRNA expression of both aging markers and apoptosis-related markers was markedly inhibited by IL-6 KO. Our results suggest that aging can be significantly reversed by IL-6 KO and that the mechanisms of this effect are related to alleviation of Mø1/Mø2 imbalance and protection against apoptosis in cardiomyocytes.


Abdominal aortic aneurysm in high-risk patients. Outcome of selective management based on size and expansion rate.
Low mortality rates for elective surgical treatment of abdominal aortic aneurysms justify an aggressive approach in most patients. However, in high-risk patients with small aneurysms and no symptoms, the decision to operate remains a delicate balance of risk and benefit. Our observations include 99 high-risk patients with asymptomatic abdominal aortic aneurysms initially measuring 3 to 6 cm in the largest transverse diameter, who have been followed 1 to 9 years (average 2.4 years) with serial echographic measurements. Elective operations were performed for aneurysmal enlargement greater than 6 cm or symptom development. An additional 11 patients with aneurysms initially greater than 6 cm, whose initial evaluation did not result in elective surgery, were also followed. Serial data documented a mean expansion rate of 0.4 cm/year for aneurysms smaller than 6 cm. Forty-one of these 99 high-risk patients with small aneurysms eventually underwent an elective resection with two deaths (4.9%). Thirty-four patients (34%) died from causes unrelated to their unoperated aneurysms, and 21 patients (21%) are alive without symptoms. Three of the 99 patients suffered aneurysm rupture and emergency operation with two deaths. Thus, of the 99 high-risk patients with small aneurysms, four have died of elective aneurysm surgery or rupture (4%). A protocol of re-echo (or computerized tomography) examination at 3-month intervals appears to define which of these high-risk patients require elective aneurysm surgery, and has limited rupture to less than 5%. Improved criteria may emerge from recent advances in high-resolution computerized tomography.


Developmental regulation of the sheep beta-lactoglobulin gene in the mammary gland of transgenic mice.
beta-Lactoglobulin (BLG) is the most abundant whey protein in sheep milk but it is not present in mouse milk. We have previously shown that transgenic mice carrying the BLG gene express it specifically in the mammary gland and secrete BLG into milk at high concentrations. Here we demonstrate that BLG transcription is correctly initiated in mice and that BLG synthesis is restricted to the secretory epithelial cells of the mammary gland. We have also determined the temporal pattern of milk protein gene expression and find that the BLG transgene is regulated coordinately with mouse beta-casein and that the patterns of regulation of BLG in mouse and sheep share some similarities.


How to Fluorescently Label the Potassium Channel: A Case in hERG.
hERG (Human ether-a-go-go-related gene) potassium channel, which plays an essential role in cardiac action potential repolarization, is responsible for inherited and druginduced long QT syndrome. Recently, the Cryo-EM structure capturing the open conformation of hERG channel was determined, thus pushing the study on hERG channel at 3.8 Å resolution. This report focuses primarily on summarizing the design rationale and application of several fluorescent probes that target hERG channels, which enables dynamic and real-time monitoring of potassium pore channel affinity to further advance the understanding of the channels.


Preparation of monoclonal antibodies to the beta A subunit of ovarian inhibin using a synthetic peptide immunogen.
Seven different synthetic peptides were prepared corresponding to regions of the beta A subunit of 32 KDa human ovarian inhibin predicted to contain possible continuous B cell epitopes. These were coupled to tuberculin as a carrier and used to immunize mice previously given a priming dose of human tuberculosis vaccine. Only one of these peptides, corresponding to sequence 82-114, consistently gave good titres of antibodies reactive with intact 32 KDa bovine inhibin by ELISA. From one of the mice immunized with this peptide, six stable hybridomas were prepared. Five of these secreted an IgM and the sixth (clone E4) secreted an IgG2b antibody. Immunoblotting experiments on follicular fluid concentrates, after treatment with sodium dodecyl sulphate and mercaptoethanol, showed strong reactivity of the antibody from clone E4 only with bands of about 13 kDa and 58 kDa corresponding to forms of the beta A subunit previously described. This monoclonal antibody, and an antibody to the alpha subunit previously made in this laboratory using synthetic peptides, should prove useful reagents for further study of inhibins and activins.


Site-specific fluorescent labeling to visualize membrane translocation of a myristoyl switch protein.
Fluorescence approaches have been widely used for elucidating the dynamics of protein-membrane interactions in cells and model systems. However, non-specific multi-site fluorescent labeling often results in a loss of native structure and function, and single cysteine labeling is not feasible when native cysteines are required to support a protein's folding or catalytic activity. Here, we develop a method using genetic incorporation of non-natural amino acids and bio-orthogonal chemistry to site-specifically label with a single fluorescent small molecule or protein the myristoyl-switch protein recoverin, which is involved in rhodopsin-mediated signaling in mammalian visual sensory neurons. We demonstrate reversible Ca(2+)-responsive translocation of labeled recoverin to membranes and show that recoverin favors membranes with negative curvature and high lipid fluidity in complex heterogeneous membranes, which confers spatio-temporal control over down-stream signaling events. The site-specific orthogonal labeling technique is promising for structural, dynamical, and functional studies of many lipid-anchored membrane protein switches.


A Phenomenological Approach to Study Mechanical Properties of Polymeric Porous Structures Processed Using Supercritical CO₂.
This work proposes a modeling of the mechanical properties of porous polymers processed by scCO₂, using a phenomenological approach. Tensile and compression tests of alginate/gelatin and cellulose acetate/graphene oxide were modeled using three hyperelastic equations, derived from strain energy functions. The proposed hyperelastic equations provide a fair good fit for mechanical behavior of the nanofibrous system alginate/gelatin (deviations lower than 10%); whereas, due to the presence of the solid in the polymer network, a four-parameter model must be used to fit the composite cellulose acetate/graphene oxide behavior. Larger deviations from the experimental data were observed for the system cellulose acetate/graphene oxide because of its microporous structure. A finite element method was, then, proposed to model both systems; it allowed a realistic description of observable displacements and effective stresses. The results indicate that materials processed using scCO₂, when submitted to large stresses, do not obey Hooke´s law and must be considered as hyperelastic.


[Effect of maximum anchorage extraction on upper airway in adolescent patients with bimaxillary protrusion].
To investigate the effect of maximum anchorage on upper airway in adolescent patients with bimaxillary protrusion and extraction from three-dimensional direction on cone-beam computed tomography(CBCT). A retrospective evaluation was carried out on pre- and post-orthodontic treatment records of 50 adolescents with bimaxillary protrusion treated by maximum anchorage extraction. The changes on upper airway pre- and post-treatment were measured. The data were analyzed using SPSS 17.0 software package. It was found that the volume, mean cross-section area, minimum cross-section area of oropharyngeal and hypopharyngeal airway were significantly decreased after treatment (P<0.001). The cross-section morphology of oropharyngeal and hypopharyngeal airway showed more ellipsoid(P<0.001). Pearson's correlation analysis indicated that the lower incisor retraction and hyoid retraction was correlated with mean cross-section area of hypopharyngeal airway (P<0.05). There was significant correlation in the upper and lower incisor retraction and hyoid retraction(P<0.01). The upper airway dimension should be given serious consideration on the decision when closing the space with maximum anchorage in adolescent patients.


Trichomycete fungi (Zygomycota) associated with the digestive tract of Simulium goeldii Cerqueira & Nunes de Mello and Simulium ulyssesi (Py-Daniel & Coscarón) (Diptera: Simuliidae) larvae, in Central Amazônia, Brazil.
Trichomycete fungi are associated with digestive tract of black fly larvae. These fungi have not been studied in Brazil, knowing them and their relationship with black flies could be an alternative to control vector populations. The objectives of this study were to survey the Trichomycete fungi associated with larvae of Simulium goeldii and S. ulyssesi, and to determine if there is specificity and/or difference in the infection rates in these species. Black flies were collected in Central Amazônia, Brazil. Three genera of Trichomycetes were found: Harpella, Genistellospora and Smittium. All these fungi were common to both black fly species.


Complex genetic regulation of bone mineral density and insulin-like growth factor-I in C57BL/6J-Chr #A/J/NaJ chromosome substitution strains.
Low bone mineral density (BMD) is a phenotype associated with osteoporosis and increased risk of fracture. Since 60-80% of variation in BMD is associated with genetic factors, we used the novel approach of chromosome substitution strains (CSS) to identify chromosomes that harbor genes that regulate BMD. Specifically, we evaluated 24 wk old C57BL/6J-Chr #(A/J)/NaJ CSS (n = 7 to 18) in which each chromosome in the host C57BL/6J strain is replaced by a corresponding chromosome from the donor A/J strain (19 autosomes, X, Y). We determined several A/J chromosomes contribute to body weight (BW), percent body fat (BF), whole body areal BMD (aBMD), and serum insulin-like growth factor (IGF)-I in both a positive and negative manner when compared with C57BL/6J. Specifically, C57BL/6J-Chr 5(A/J)/NaJ (B.A-5) (males) and B.A-13 (females) contributed to increased BW, whereas B.A-3, 4, 8, 9, 12, and 18 (males) and B.A-3, 4, and 11 (females) contributed to reduced BW. B.A-5 (males) and B.A-13 (females) contributed to increased BF, whereas B.A-12 (males) and B.A-3, 4, 10, and 11 (females) contributed to reduced BF. B.A-14 (females) contributed to increased aBMD and B.A-1, 2, 6, 9, 10, and 18 (males) and B.A-8, 9, and 10 (females) contributed to reduced aBMD. To determine if similar chromosomes regulate aBMD and IGF-I, we determined serum concentrations of IGF-I. B.A-14 and Y (males) and B.A-6 (females) contributed to increased IGF-I and male B.A-3 and female B.A-8 contributed to reduced IGF-I. Overall, we identified several sex-dependent and -independent chromosomes that regulate aBMD and IGF-I in adult mice.


[Acne vulgaris. The role of hormones].
Peripherally active androgens control cellular functions by binding to androgen receptors. Follicular keratinocytes and sebocytes are target cells for androgens, which, directly or indirectly, stimulate keratinocyte proliferation and the volume of sebaceous glands as well as the sebum secretion rate. Acne often begins with the adrenarche, namely with the up- regulation of adrenal synthesis of dehydroepiandrosterone sulfate, a hormone that is upstream to testosterone. The majority of acne patients exhibit normal levels of circulating androgens, while sebaceous glands from acne regions exhibit a stronger sensitivity to androgens than the sebaceous glands from other parts of the body. Hormone-like active lipids, hormones in diet and neuropeptides may also play a role in the development of acne lesions. The target of antiandrogen treatment of the skin is mainly the sebaceous gland and the primary effect is sebostasis.


European Society of Endocrine Surgeons (ESES) and European Network for the Study of Adrenal Tumours (ENSAT) recommendations for the surgical management of adrenocortical carcinoma.
Radical surgery provides the best chance of cure for adrenocortical carcinoma (ACC), but perioperative surgical care for these patients is yet to be standardized. A working group appointed jointly by ENSAT and ESES used Delphi methodology to produce evidence-based recommendations for the perioperative surgical care of patients with ACC. Papers were retrieved from electronic databases. Evidence and recommendations were classified according to the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) system, and were discussed until consensus was reached within the group. Twenty-five recommendations for the perioperative surgical care of patients with ACC were formulated. The quality of evidence is low owing to the rarity of the disease and the lack of prospective surgical trials. Multi-institutional prospective cohort studies and prospective RCTs are urgently needed and should be strongly encouraged. The present evidence-based recommendations provide comprehensive advice on the optimal perioperative care for patients undergoing surgery for ACC.


Socioeconomic and drug use determinants of smoking status in an urban adult population of Black African descent.
We examined the influence of socioeconomics and drug use on current smokers (N = 137) and nonsmokers (N = 143) from an urban adult population of Black African descent. Median participant age was 33 years (range = 20-59). Smokers consumed a median of eight cigarettes/day (range = 0-35). Interestingly, 86% smoked fewer than 15 cigarettes/day and only 8% smoked menthol cigarettes. Socioeconomic and drug use variables significantly associated with smoking status in univariate analyses were included in a multiple logistic regression model that controlled for gender and age. Compared with nonsmokers, smokers were less likely to be university educated, more likely to be divorced, separated, or widowed, more likely to be current alcohol users, and more likely to be current marijuana users. Unexpectedly, household income and employment status were not associated with smoking status. Among current alcohol users, smokers consumed three times the number of drinks per month that nonsmokers consumed (p<.001). Among current marijuana users, smokers consumed more than five times the number of joints per month that nonsmokers consumed (p<.001). Overall, lower education levels, divorce, and alcohol and marijuana use were significantly associated with increased likelihood of smoking among this population.


Immunofluorescent analysis of extracellular matrix (ECM) components in glomeruli of the hepatic glomerulosclerosis.
An immunofluorescence study was carried out to evaluate alterations in the distribution and/or intensity of extracellular matrix (ECM) components, immunoglobulins and complement (C3) in the glomeruli of 7 autopsy patients with hepatic glomerulosclerosis. As compared with the findings in normal renal tissues, an increase in type IV collagen, laminin and fibronectin was observed in expanded glomerular mesangial areas and along glomerular capillary walls. Depositions of IgA, mainly IgA1, and C3 in glomeruli were in parallel with those of the ECM components. These ECM components were markedly decreased in glomeruli showing global sclerosis. In contrast, type I collagen was observed at central portions of the sclerotic glomeruli. There was an increase in the type I collagen along Bowman's capsules, especially at the sites of capsular adhesion and crescent formation. It appears that hyperproduction and/or infiltration of glomerular ECM components and interstitial collagen is closely linked to the progression of glomerular sclerosis in patients with liver diseases.


Force characteristics of in vivo tissue-engineered myocardial constructs using varying cell seeding densities.
Experiments have been successfully performed culminating in functional, vascularized, three-dimensional cardiac muscle tissue. Past experience in tissue engineering has led us to the understanding that cell seeding density plays a critical role in the formation and function of both in vitro and in vivo engineered tissues. Therefore, to improve upon the mechanics of this model and to facilitate the formation of myocardial tissue with improved functional performance, we sought to optimize the seeding density of cardiomyocytes in these constructs. Neonatal cardiac myocytes were isolated from 2-day-old Fischer 344 rat hearts. Silicone chambers containing fibrin gel were seeded with varying numbers of cardiac cells (1, 5, 10, and 20 million). Control chambers were prepared using fibrin gel alone. All of the chambers were then implanted around the femoral vessels of isogenic rats. Six constructs per cell seeding density group were implanted. Histological and immunohistochemical evaluation was performed via hematoxylin and eosin, von Gieson, and alpha-sarcomeric actin staining protocols. Linear contractile force measurements were obtained for each construct following 4 weeks of in vivo implantation. After an implantation period of 4 weeks, the newly formed cardiac constructs contained within the chambers were harvested. The femoral vessels within the constructs were found to be patent in all cases. With direct electrical stimulation, the constructs were able to generate an average active force that varied depending on their seeding density. Constructs with seeding densities of 1, 5, 10, and 20 million cells produced an average active force of 208, 241, 151, and 108 microN, respectively. The control constructs did not generate any active force on electrical stimulation. This study demonstrates the in vivo survival, vascularization, organization, and function of transplanted myocardial cells. It is also apparent that cell seeding density plays a direct role in the force generation and mechanical properties of these engineered constructs. Among different groups using varying cell seeding densities, we found that the group with 5 million cells generated maximum active force.


Nanotechnology, Green Synthesis and Biological Activity Application of Zinc Oxide Nanoparticles Incorporated Argemone Mxicana Leaf Extract.
Nanomaterial is a rapidly growing area that is used to create a variety of new materials and nanotechnology applications from medical, pharmaceuticals, chemical, mechanical, electronics and several environmental industries including physical, chemical and biological nanoparticles are very important in our daily life. Nanoparticles with leaf extract from the healthy plant are important in the area of research using biosynthesis methods. Because of it's used as an environmentally ecofriendly, other than traditional physical and chemical strategies. In particular, biologically synthesized nanoparticles have become a key branch of nanotechnology. The present work presents a synthesis of zinc oxide nanoparticles using an extract from the Argemone leaf Mexicana. Biosynthetic nanoparticles are characterized by X-ray diffraction (XRD), Ultraviolet visible (UV-vis) spectroscopy analysis, a Fourier Transform Infrared Spectroscopy analysis (FTIR) and a scanning electron microcopy (SEM), X-ray analysis with dispersive energy (EDAX). XRD is used to examine the crystalline size of zinc oxide nanoparticles. The FTIR test consists in providing evidence of the presence of targeted teams. UV is used for optical properties and calculates the energy of the bandwidth slot. The scanning microscope emission reveals the morphology of the surface and the energy dispersive X-ray analysis confirms the basic composition of zinc oxide nanoparticles. It is found that zinc nanoparticles are capable of achieving high anti-fungal efficacy and therefore have a high potential antimicrobial activity of ZnO NPs, like antibacterial and high antioxidant. Zinc Oxide nanoparticles from the Argemone Mexicana leaf extract have several antimicrobial applications, such as medical specialty, cosmetics, food, biotechnology, nano medicine and drug delivery system. ZnO nanoparticles are important because they provide many practical applications in industry. The most important use of nanoparticles of ZnO would be strong antibacterial and antioxidant activity with a simple and efficient biosynthesis method may be used for future work applications.


Prebiotic fiber increases hepatic acetyl CoA carboxylase phosphorylation and suppresses glucose-dependent insulinotropic polypeptide secretion more effectively when used with metformin in obese rats.
Independently, metformin (MET) and the prebiotic, oligofructose (OFS), have been shown to increase glucagon-like peptide (GLP-1) secretion. Our objective was to determine whether using OFS as an adjunct with MET augments GLP-1 secretion in obese rats. Male, diet-induced obese Sprague Dawley rats were randomized to: 1) high-fat/-sucrose diet [HFHS; control (C); 20% fat, 50% sucrose wt:wt]; 2) HFHS+10% OFS (OFS); 3) HFHS + MET [300 mg/kg/d (MET)]; 4) HFHS+10% OFS+MET (OFS+MET). Body composition, glycemia, satiety hormones, and mechanisms related to dipeptidyl peptidase 4 (DPP4) activity in plasma, hepatic AMP-activated protein kinase (AMPK; Western blots), and gut microbiota (qPCR) were examined. Direct effects of MET and SCFA were examined in human enteroendocrine cells. The interaction between OFS and MET affected fat mass, hepatic TG, secretion of glucose-dependent insulinotropic polypeptide (GIP) and leptin, and AMPKα2 mRNA and phosphorylated acetyl CoA carboxylase (pACC) levels (P < 0.05). Combined, OFS and MET reduced GIP secretion to a greater extent than either treatment alone (P < 0.05). The hepatic pACC level was increased by OFS+MET by at least 50% above all other treatments, which did not differ from each other (P < 0.05). OFS decreased plasma DPP4 activity (P < 0.001). Cecal Bifidobacteria (P < 0.001) were markedly increased and C. leptum decreased (P < 0.001) with OFS consumption. In human enteroendocrine cells, the interaction between MET and SCFA affected GLP-1 secretion (P < 0.04) but was not associated with higher GLP-1 than the highest individual doses. In conclusion, the combined actions of OFS and MET were associated with important interaction effects that have the potential to improve metabolic outcomes associated with obesity.


Effects of drugs on glucose measurements with handheld glucose meters and a portable glucose analyzer.
Thirty drugs used primarily in critical care and hospital settings were tested in vitro to observe interference on glucose measurements with 6 hand-held glucose meters and a portable glucose analyzer. Paired differences of glucose measurements between drug-spiked samples and unspiked control samples were calculated to determine bias. A criterion of +/- 6 mg/dL was used as the cutoff for interference. Ascorbic acid interfered with the measurements on all glucose devices evaluated. Acetaminophen, dopamine, and mannitol interfered with glucose measurements on some devices. Dose-response relationships help assessment of drug interference in clinical use. High dosages of these drugs may be given to critically ill patients or self-administered by patients without medical supervision. Package inserts for the glucose devices may not provide adequate warning information. Hence, we recommend that clinicians choose glucose devices carefully and interpret results cautiously when glucose measurements are performed during or after drug interventions.


Resistance-associated mutations in HIV-1 among patients failing first-line antiretroviral therapy.
Thirty-five HIV-1 infected patients showing clinical and/or immunological failure to first line antiretroviral therapy (ART) according to WHO criteria were recruited from the ART center of Lok Nayak Hospital, New Delhi to detect the presence of resistance-mutations in reverse transcriptase (RT) and protease (PR) region of pol gene of HIV-1. Plasma viral load (PVL) was estimated. HIV-1 pol gene region encoding complete protease and reverse transcriptase (codons; 1-232 to 1-242) was reverse transcribed, followed by nested PCR. The PCR product was sequenced and analyzed. Plasma samples from 94.3% of patients with PVL >log(10) 3.0 c/mL could be amplified and analyzed. Virologic failure was detected in 65.7% of patients according to WHO criteria (PVL >log(10) 4.0). All patients were found to be infected with subtype C. One or more resistance-mutations were observed among 90.9% of study sequences. Nucleoside reverse transcriptase inhibitor (NRTI) resistance mutations were seen among all patients, with M184V and thymidine analogue mutations (TAM) being most frequently detected (75.6% and 72.7%, respectively). Nonnucleoside reverse transcriptase inhibitor (NNRTI) resistance-mutations were detected in 63.6% of sequences, of which Y181C/I (47.6%), K103N (33.3%) and G190S (28.6%) are the most common. None of the sequences showed major protease inhibitors (PIs) resistance mutation. High prevalence of NRTI and NNRTI drug resistance mutations among the study participants warrants the use of genotypic resistance testing to prevent accumulation of resistance mutations, which would limit future treatment options.


Novel Passive Two-Stage Magnetic Targeting Devices for Distal Locking of Interlocking Nails.
Interlocking nailing is a common surgical operation to stabilize fractures in long bones. One of the difficult parts of the surgery is how to locate the position and direction of a screw hole on the interlocking nail, which is invisible to the naked eye after insertion of the nail into the medullary canal. Here, we propose a novel two-stage targeting process using two passive magnetic devices to locate the position and direction of the screw hole without radiation for the locking screw procedure. This involves a ring-shape positioning magnet inside the nail to generate a magnetic field for targeting. From the accuracy test results of these two-stage targeting devices, the search region can be identified in less than 20 seconds by the 1st-stage targeting device, while the total targeting time to locate the drilling position and direction takes less than 4 minutes, with 100% successful rate in 50 attempts. The drilling test further combines the two-stage targeting process and drilling process on the swine tibia, and it is shown that a 100% successful rate is achieved in all 10 attempts, where the total time needed is less than 5 minutes.


Non-expanded adipose stromal vascular fraction cell therapy for multiple sclerosis.
The stromal vascular fraction (SVF) of adipose tissue is known to contain mesenchymal stem cells (MSC), T regulatory cells, endothelial precursor cells, preadipocytes, as well as anti-inflammatory M2 macrophages. Safety of autologous adipose tissue implantation is supported by extensive use of this procedure in cosmetic surgery, as well as by ongoing studies using in vitro expanded adipose derived MSC. Equine and canine studies demonstrating anti-inflammatory and regenerative effects of non-expanded SVF cells have yielded promising results. Although non-expanded SVF cells have been used successfully in accelerating healing of Crohn's fistulas, to our knowledge clinical use of these cells for systemic immune modulation has not been reported. In this communication we discuss the rationale for use of autologous SVF in treatment of multiple sclerosis and describe our experiences with three patients. Based on this rationale and initial experiences, we propose controlled trials of autologous SVF in various inflammatory conditions.


Parallel technique of endobronchial balloon catheter tamponade for transient alleviation of massive hemoptysis.
Endobronchial balloon tamponade is an alternative method that can be used to control massive hemoptysis. Several different techniques have been used for this purpose. We describe a new parallel method, in which biopsy forceps introduced through a bronchoscope channel is used to grasp a balloon catheter. As the bronchoscope is advanced to the bleeding site, the balloon catheter is pulled into position, and subsequently inflated. There are several advantages of this technique. It needs no specialized catheter or guide wire, the procedure is relatively easy to perform, and applicable to other purposes such as introduction of an additional suction catheter.


Understanding and Improving the Burden and Unmet Needs of Informal Caregivers of Homebound Patients Enrolled in a Home-Based Primary Care Program.
The growing homebound population is heavily reliant on informal caregivers, who are increasingly burdened by their roles. This study describes informal caregivers of the homebound who remain caregivers at a 9-month follow-up and examines the impact of a home-based primary care (HBPC) program on caregiver burden and unmet needs using a prospective design with a pre-post intervention assessment. Informal caregivers of the urban homebound are similar to caregivers of other populations and have a broad range of unmet needs. The intervention described is the regular provision of multidisciplinary HBPC. Hundred fourteen caregivers of newly admitted patients complete a baseline interview. For the 56 caregivers who complete a 9-month follow-up interview, participation in HBPC is associated with a statistically significant decrease in overall caregiver burden. At 9 months, caregivers demonstrate an overall decrease in unmet needs, with a statistically significant decrease across two measured domains. These results suggest that the regular provision of multidisciplinary care in the home can mitigate the deleterious impact of informal caregiving.


Rolling adhesion of human NK cells to porcine endothelial cells mainly relies on CD49d-CD106 interactions.
Acute vascular rejection in pig-to-primate xenotransplantation involves recognition and damage of porcine (po) endothelial cells (EC) by human (hu) leukocytes, probably including natural killer (NK) cells. To study such interactions we analyzed rolling and static adhesion of hu NK cells to po EC. The effects of blocking hu and po adhesion molecules on the adhesion hu NK cells to po EC monolayers was analyzed under shear stress (10 min, 37 degrees C, 0.7 dynes/cm2) or under static conditions (10 min, 37 degrees C). All used cell populations were phenotypically characterized by flow cytometry. Blocking of CD106 on po EC or its ligand CD49d on hu NK cells decreased rolling adhesion of both fresh and activated hu NK cells by more than 75%. Masking of CD62L on fresh but not activated hu NK resulted in a 44% decrease in rolling adhesion, in line with the diminished cell surface expression of CD62L upon activation. Antibodies to CD31, CD54, CD62E, and CD62P on EC or CD11a, CD18, and CD162 on NK cells had only minor effects on rolling adhesion. The adhesion of the FcgammaRIII- hu NK cell line NK92 to po EC was inhibited by 95% after masking po CD106 whereas antibodies to po CD31, CD54, CD62E, or CD62P had no effect, thereby excluding effects of Fc-receptor-dependent binding of hu NK cells to po EC. Static adhesion of activated NK cells was reduced by approximately 60% by blocking either CD49d or CD106, by 47% by blocking CD11a, and by 82% upon simultaneous blocking of CD11a and CD49d. Interactions between hu CD49d and po CD106 are crucial for both rolling and firm adhesion of hu NK cells to po EC and thus represent attractive targets for specific therapeutic interventions to prevent NK cell-mediated responses against po xenografts.


[Inhibitory effects of BIO-1211 on bronchoconstriction and neutrophil adhesion in rats].
To determine the inhibitory effects of BIO-1211, a very late antigen-4 (vla-4) antagonist, on bronchoconstriction and neutrophil adhesion in rats. For evaluating ovalbumin-induced bronchoconstriction in the sensitized rats, the changes in lung resistance (RL) and lung dynamic compliance (C(dyn)) were determined after antigen challenge. Neutrophils from the rats were used to determine fibronectin and serum-induced cell adhesion. The effect of BIO-1211 on wheezing was determined after inhalation of histamine and acetylcholine in guinea pigs. BIO-1211 aerosol at 1, 3 and 10 mg/ml significantly inhibited the changes in lung resistance and lung dynamic compliance after antigen challenge in the sensitized rats in a dose-dependent manner. BIO-1211 at 25, 50, 100 and 200 microgram/ml inhibited the fibronectin-induced neutrophil adhesion by 23.5%, 24.6%, 61.4% and 58.1%, respectively, and serum-induced adhesion by 29.9%, 35.9%, 35.3% and 15.4%, respectively. Inhalation of 10 mg/ml BIO-1211 did not show any protection against histamine and acetylcholine-induced bronchoconstriction. BIO-1211 inhibits bronchoconstriction and neutrophil adhesion, which may be associated with its effect against bronchoconstriction in rats.


Diagnostics to support the eradication of yaws-Development of two target product profiles.
Yaws is targeted for eradication by 2030, using a strategy based on mass drug administration (MDA) with azithromycin. New diagnostics are needed to aid eradication. Serology is currently the mainstay for yaws diagnosis; however, inaccuracies associated with current serological tests makes it difficult to fully assess the need for and impact of eradication campaigns using these tools. Under the recommendation of the WHO Diagnostic Technical Advisory Group (DTAG) for Neglected Tropical Diseases(NTDs), a working group was assembled and tasked with agreeing on priority use cases for developing target product profiles (TPPs) for new diagnostics tools. The working group convened three times and established two use cases: identifying a single case of yaws and detecting azithromycin resistance. One subgroup assessed the current diagnostic landscape for yaws and a second subgroup determined the test requirements for both use cases. Draft TPPs were sent out for input from stakeholders and experts. Both TPPs considered the following parameters: product use, design, performance, configuration, cost, access and equity. To identify a single case of yaws, the test should be able to detect an analyte which confirms an active infection with at least 95% sensitivity and 99.9% specificity. The high specificity was deemed important to avoid a high false positive rate which could result in unnecessary continuation or initiation of MDA campaigns. If used in settings where the number of suspected cases is low, further testing could be considered to compensate for imperfect sensitivity and to improve specificity. The test to detect azithromycin resistance should be able to detect known genetic resistance mutations with a minimum sensitivity and specificity of 95%, with the caveat that all patients with suspected treatment failure should be treated as having resistant yaws and offered alternative treatment. The TPPs developed will provide test developers with guidance to ensure that novel diagnostic tests meet identified public health needs.


Isolation of carboxylester lipase (CEL) isoenzymes from Candida rugosa and identification of the corresponding genes.
The yeast Candida rugosa produces extracellular lipases which are widely used for industrial purposes. A commercial lipase preparation from this yeast can be separated into several isoenzymes which differ in carbohydrate content, isolelectric point, substrate specificity, and primary sequence. We have here purified and characterized three lipases, which also hydrolyze p-nitrophenyl esters, from a commercial preparation of this yeast. These three carboxylester lipases (CELs) elute differently on hydrophobic interaction chromatography, and have different carbohydrate contents and substrate specificities. Sequence analysis of their amino termini and peptides generated by LysC treatment showed that CEL-1 and CEL-3 probably have identical primary structure while CEL-2 was proven to be a different enzyme. Sequence comparison showed that both CEL-1 and CEL-3 are products of the LIP1 gene and that CEL-2 is the gene product of LIP2, cloned by Longhi et al. (Biochim. Biophys. Acta 1131, 227-232, 1992).


Highly Effective Antibacterial Vesicles Based on Peptide-Mimetic Alternating Copolymers for Bone Repair.
It is an important challenge for bone repair to effectively deliver growth factors and at the same time to prevent and cure inflammation without obvious pathogen resistance. We designed a kind of antibacterial peptide-mimetic alternating copolymers (PMACs) to effectively inhibit and kill both Gram-positive and Gram-negative bacteria. The minimum inhibition concentrations (MICs) of the PMACs against E. coli and S. aureus are 8.0 μg/mL, which are much lower than that of antibacterial peptides synthesized by other methods such as widely used ring-opening polymerization of N-carboxyanhydride. Furthermore, the PMACs can self-assemble into polymer vesicles (polymersomes) in pure water with low cytotoxicity (IC50 > 1000 μg/mL), which can encapsulate growth factors in aqueous solution and release them during long-term antibacterial process for facilitating bone repair. We also find that the alternating structure is essential for the excellent antibacterial activity. The in vivo tests in rabbits confirmed that the growth-factor-encapsulated antibacterial vesicles have better bone repair ability compared with control groups without antibacterial vesicles. Overall, we have provided a novel method for designing PMAC-based highly effective intrinsically antibacterial vesicles that may have promising biomedical applications in the future.


Inhibition of Vpx-Mediated SAMHD1 and Vpr-Mediated Host Helicase Transcription Factor Degradation by Selective Disruption of Viral CRL4 (DCAF1) E3 Ubiquitin Ligase Assembly.
The lentiviral accessory proteins Vpx and Vpr are known to utilize CRL4 (DCAF1) E3 ligase to induce the degradation of the host restriction factor SAMHD1 or host helicase transcription factor (HLTF), respectively. Selective disruption of viral CRL4 (DCAF1) E3 ligase could be a promising antiviral strategy. Recently, we have determined that posttranslational modification (neddylation) of Cullin-4 is required for the activation of Vpx-CRL4 (DCAF1) E3 ligase. However, the mechanism of Vpx/Vpr-CRL4 (DCAF1) E3 ligase assembly is still poorly understood. Here, we report that zinc coordination is an important regulator of Vpx-CRL4 E3 ligase assembly. Residues in a conserved zinc-binding motif of Vpx were essential for the recruitment of the CRL4 (DCAF1) E3 complex and Vpx-induced SAMHD1 degradation. Importantly, altering the intracellular zinc concentration by treatment with the zinc chelator N,N,N'-tetrakis-(2'-pyridylmethyl)ethylenediamine (TPEN) potently blocked Vpx-mediated SAMHD1 degradation and inhibited wild-type SIVmac (simian immunodeficiency virus of macaques) infection of myeloid cells, even in the presence of Vpx. TPEN selectively inhibited Vpx and DCAF1 binding but not the Vpx-SAMHD1 interaction or Vpx virion packaging. Moreover, we have shown that zinc coordination is also important for the assembly of the HIV-1 Vpr-CRL4 E3 ligase. In particular, Vpr zinc-binding motif mutation or TPEN treatment efficiently inhibited Vpr-CRL4 (DCAF1) E3 ligase assembly and Vpr-mediated HLTF degradation or Vpr-induced G2 cell cycle arrest. Collectively, our study sheds light on a conserved strategy by the viral proteins Vpx and Vpr to recruit host CRL4 (DCAF1) E3 ligase, which represents a target for novel anti-human immunodeficiency virus (HIV) drug development.IMPORTANCE The Vpr and its paralog Vpx are accessory proteins encoded by different human immunodeficiency virus (HIV)/simian immunodeficiency virus (SIV) lentiviruses. To facilitate viral replication, Vpx has evolved to induce SAMHD1 degradation and Vpr to mediate HLTF degradation. Both Vpx and Vpr perform their functions by recruiting CRL4 (DCAF1) E3 ligase. In this study, we demonstrate that the assembly of the Vpx- or Vpr-CRL4 E3 ligase requires a highly conserved zinc-binding motif. This motif is specifically required for the DCAF1 interaction but not for the interaction of Vpx or Vpr with its substrate. Selective disruption of Vpx- or Vpr-CRL4 E3 ligase function was achieved by zinc sequestration using N,N,N'-tetrakis-(2'-pyridylmethyl)ethylenediamine (TPEN). At the same time, zinc sequestration had no effect on zinc-dependent cellular protein functions. Therefore, information obtained from this study may be important for novel anti-HIV drug development.


Theoretical analysis of bonding in N-heterocyclic carbene-rhodium complexes.
The natural orbitals for chemical valence and the Ziegler-Rauk bond energy decomposition analysis were used to describe the donor/acceptor character of the N-heterocyclic carbenes (NHC)-metal bond in two groups of square-planar rhodium(I) complexes: (NHC)RhCl(cod) (1-X; cod = 1,5-cyclooctadiene) and (NHC)RhCl(CO)(2) (2-X), with a group X = H, Cl, NO(2), or CN located on the NHC ligand. The results show that the NHC-metal bond consists of the components originating from donation (sigma symmetry) and back-donation (two contributions of the pi symmetry, out-of-plane and in-plane, accompanied by one sigma-back-bonding component). The charge-flow measures from NOCV indicate that the total back-bonding contribution is of comparable importance to donation. The out-of-plane pi component contributes to ca. 50% of the total back-bonding charge-flow. The energy measures from the Ziegler-Rauk analysis show that the total back-bonding energy corresponds to ca. 40% of the orbital interaction energy. The ligand trans to NHC (CO or cod) strongly affects the back-bonding component; for the complexes 1-X, the back-donation is substantially enhanced compared to 2-X. The back-bonding component increases with an increase in the pi-withdrawing ability of X for both, 1-X and 2-X. However, this effect is relatively small. Back-bonding components of the two bonds involving the metal are strongly coupled; an increase in NHC-Rh leads to a decrease in Rh-olefin/CO(trans). The changes in the back-bonding are too small to be followed by the trends in bond energies, which are finally determined by the electrostatic and Pauli repulsion energy.


An improved staining method for rat microglial cells using the lectin from Griffonia simplicifolia (GSA I-B4).
A simple method for the lectin histochemical visualization of rat microglial cells is described. Advantages include ease of fixation of brain tissue using paraformaldehyde, and rapidity of tissue processing by vibratome sectioning. Furthermore, in addition to providing good structural preservation, the method achieves improved lectin binding, resulting in complete labeling of all microglial cells and in superior visualization of cellular processes. The lectin histochemical technique for rat microglia has the potential to be adapted to any mammalian species, and should prove valuable for neuroscientists interested in studying this glial cell type.


The combination of Lonicerae Japonicae Flos and Forsythiae Fructus herb-pair alleviated inflammation in liver fibrosis.
Objective: To explore the active components and epigenetic regulation mechanism underlying the anti-inflammatory effects of Lonicerae Japonicae Flos and Forsythiae Fructus herb-pair (LFP) in carbon tetrachloride (CCl4)-induced rat liver fibrosis. Methods: The main active ingredients and disease-related gene targets of LFP were determined using TCMSP and UniProt, and liver fibrosis disease targets were screened in the GeneCards database. A network was constructed with Cytoscape 3.8.0 and the STRING database, and potential protein functions were analyzed using bioinformatics analysis. Based on these analyses, we determined the main active ingredients of LFP and evaluated their effects in a CCl4-induced rat liver fibrosis model. Serum biochemical indices were measured using commercial kits, hepatocyte tissue damage and collagen deposition were evaluated by histopathological studies, and myofibroblast activation and inflammation were detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. High-performance liquid chromatography-mass spectrometry was performed to determine the levels of homocysteine, reduced glutathione, and oxidized glutathione, which are involved in inflammation and oxidative stress. Results: The main active components of LFP were quercetin, kaempferol, and luteolin, and its main targets were α-smooth muscle actin, cyclooxygenase-2, formyl-peptide receptor-2, prostaglandin-endoperoxide synthase 1, nuclear receptor coactivator-2, interleukinβ, tumor necrosis factor α, CXC motif chemokine ligand 14, and transforming growth factor β1. A combination of quercetin, kaempferol, and luteolin alleviated the symptoms of liver fibrosis. Conclusion: The results of this study support the role of LFP in the treatment of liver fibrosis, and reveal that LFP reduces collagen formation, inflammation, and oxidative stress. This study suggests a potential mechanism of action of LFP in the treatment of liver fibrosis.


A modified I-PCR to detect the factor VIII Inv22 for genetic diagnosis and prenatal diagnosis in haemophilia A.
To explore the effectiveness of modified inversion-polymerase chain reaction (I-PCR) to detect the factor VIII (FVIII) intron 22 inversion (Inv22) for genetic diagnosis and prenatal diagnosis in haemophilia A (HA). Both modified I-PCR and LD-PCR were applied to analyse the FVIII Inv22 for 24 patients with HA. Prenatal diagnosis was performed on six foetuses. Foetal blood samplings were carried out by cordocentesis from 22 to 26 weeks of gestation. Ten patients with FVIII Inv22 in 10 HA families were found, and the remaining 14 patients were found without the Inv22 in 19 HA families. Prenatal diagnosis confirmed that four foetuses were normal and all of them born normally. However, two foetuses had been identified as abnormal and undergone abortion. Compared with LD-PCR, modified I-PCR is more rapid and convenient for detecting the FVIII Inv22 in genetic diagnosis. It is recommended that a patient undergoes both modified I-PCR (to detect the FVIII Inv22) and biochemical assay (to measure the FVIII activity of umbilical cord blood) in prenatal diagnosis. When we have more experience, the DNA samples from chorionic villus or amniotic fluid can be analysed for prenatal diagnosis using the modified I-PCR alone.


[Study on the extraction recovery of ginsenoside Re in plasma by different solid-phase cartridges].
To optimize the solid-phase extraction method by comparison of the extraction recovery of ginsenoside Re plasma samples. After extracted by different solid-phase cartridges with water, acetonitrile, and different content methanol elution, the plasma samples were analyzed on an Zorbax SB-C18 column with acetonitrile-water gradient elution. From the recovery achieved, the best solid phase cartridge was found. This method consists of using 40% methanol as the wash solvent, and 80% methanol for the elution. Among the three kinds of solid-phase being tested, Waters Oasis HLB cartridge was found to be the best one. The average extraction recovery of the Waters Oasis HLB cartridges was between 103%-113%, it can be used in the analysis of ginsenoside Re in plasma samples.


[Analysis on clinical characteristics and prognosis of patients with mitral valve prolapse].
Objective: To evaluate the clinical characteristics and prognosis of patients with mitral valve prolapse (MVP). Methods: We retrospectively analyzed the clinical characteristics and prognosis of 148 MVP patients who underwent mitral valve surgery in Fuwai hospital from January 2012 to December 2015.The patients were divided into mucoid degeneration group (52 cases) and without mucoid degeneration group(19 cases) according to pathological examination of leaflets and chordate. Results: The clinical symptoms of MVP patients included dyspnea (59.5%(88/148)), chest distress and pain (52.7%(78/148)), and palpitations (36.5% (54/148)). Mitral valve repair was performed in 144 cases (97.3%), and mitral valve replacement was performed in 4 cases (2.7%). Posterior leaflet prolapse was the most common form of MVP (68.9%, 102/148). Pathological examination revealed myxomatous degeneration in 73.2% patients (52/71), fibrosis in 8.5% patients (6/71), and fibrinoid necrosis in 8.5% patients (6/71). Patients with mucoid degeneration had less atrial fibrillation before surgery (5.8%(3/52) vs. 42.1%(8/19), P<0.01), smaller preoperative left atrium diameter ((43.2±6.5) mm vs. (48.2±8.9) mm, P<0.05), more posterior leaflet prolapse (94.2%(49/52) vs. 63.2%(12/19), P<0.01), redundant chordae (26.9%(14/52) vs. 0, P<0.05) and leaflet thickening (76.9%(40/52) vs. 52.6%(10/19), P<0.05) when compared with patients without mucoid degeneration.Echocardiography examination at the postoperative follow-up of 39.0(22.3, 57.0) months revealed smaller left atrium diameter((38.5±7.1) mm vs. (45.3±8.3) mm, P<0.01), left ventricular end-diastolic diameter ((48.9±6.2) mm vs. (57.5±7.6) mm, P<0.01), reduced left ventricular ejection fraction ((61.2±7.1)% vs. (65.1±6.2)%, P<0.01) and less moderate or severe mitral regurgitation (1.4%(2/148) vs. 100.0%(148/148), P<0.01) compared with the corresponding preoperative values. Conclusions: Dyspnea is the main symptom, and mucoid degeneration characterized by redundant chordae and leaflet thickening are the main pathological features of MVP patients.The surgical treatment of MVP patients is related with satisfactory outcome results.


Effect of in vitro X-irradiation on lymphocyte and granulocyte function.
Granulocyte transfusions have been complicated by graft-versus-host disease (GVHD) in the recipients. This risk can be eliminated by irradiation of the cell product. The effect of in vitro irradiation on elements of lymphocyte and granulocyte function was therefore studied in order to determine the dose of irradiation which blocked lymphocyte function without affecting the function of granulocytes. Lymphocyte blast transformation after stimulation with mitogens was reduced by 90% after irradiation with 1500 rad and by 97% after 5 000 rad. The response to microbial agents and allogeneic cells was far more radio-sensitive, being completely abolished after irradiation with 1 000 and 500 rad, respectively. Mobility was the function of polymorphonuclear leucocytes (PMN) most affected by irradiation, being slightly but significantly reduced after irradiation with 10 000-20 000 rad. The bactericidal activity was reduced only after irradiation with 40 000 rad or more, while the hexose monophosphate shunt activity and the myeloperoxidase activity were largely unaffected by irradiation with doses of up to 120 000 rad. Hence the results indicate that the irradiation of leucocytes intended for transfusion with a dose of 2 000 rad is likely to prevent GVHD without causing any apparent damage to the PMN.


Neuropeptide Y (NPY)-induced suppression of activity in the rat: evidence for NPY receptor heterogeneity and for interaction with alpha-adrenoceptors.
The receptor mechanisms mediating the neuropeptide Y (NPY)-induced suppression of behavioural activity have been examined in the rat. The interaction of NPY with central noradrenergic mechanisms was also studied. The non-selective alpha-adrenoceptor antagonist, phentolamine (15-60 nmol intracerebroventricularly, i.c.v.), caused a dose-related antagonism (up to 50%) of the NPY-induced suppression of activity. The selective alpha 2-adrenoceptor antagonist, idazoxan (0.125 mg/kg intraperitoneally, i.p.), was even more effective, while the selective alpha 1-adrenoceptor antagonist, prazosin, was without effect. In addition, we examined whether the recently postulated subdivision of peripheral NPY receptors was also applicable to the brain. The ability of the C-terminal 13-36 amino acid fragment of NPY (postulated to activate NPY-Y2 receptors) to reproduce the effects of the full molecule (postulated to activate both NPY-Y1 and -Y2 receptors) was tested. NPY-(13-36) (0.4-10.0 nmol i.c.v.) failed to produce any suppression of activity. On the contrary, it produced an increase in locomotor activity and rearings at low doses. This effect was not blocked by phentolamine. We conclude that the NPY-induced suppression of activity is produced to a large extent by modulation of alpha 2-adrenergic transmission. Our results also provide evidence for heterogeneity among the central NPY receptors, with the NPY-induced suppression of activity being mediated by the NPY-Y1 receptor subtype.


Dietary arginine affects energy metabolism through polyamine turnover in juvenile Atlantic salmon (Salmo salar).
In the present study, quadruplicate groups of juvenile Atlantic salmon (Salmo salar) were fed plant protein-based diets with increasing arginine inclusions (range 28·8-37·4 g/kg DM) to investigate whether arginine supplementation affects growth and lipid accumulation through an elevated polyamine turnover. Dietary lysine was held at a constant concentration, just below the requirement. All other amino acids were balanced and equal in the diets. Arginine supplementation increased protein and fat accretion, without affecting the hepatosomatic or visceralsomatic indices. Dietary arginine correlated with putrescine in the liver (R 0·78, P= 0·01) and with ornithine in the muscle, liver and plasma (P= 0·0002, 0·003 and 0·0002, respectively). The mRNA of ornithine decarboxylase, the enzyme producing putrescine, was up-regulated in the white adipose tissue of fish fed the high-arginine inclusion compared with those fed the low-arginine diet. Concomitantly, spermidine/spermine-(N1)-acetyltransferase, the rate-limiting enzyme for polyamine turnover that consumes acetyl-CoA, showed an increased activity in the liver of fish fed the arginine-supplemented diets. In addition, lower acetyl-CoA concentrations were observed in the liver of fish fed the high-arginine diet, while ATP, which is used in the process of synthesising spermidine and spermine, did not show a similar trend. Gene expression of the rate-limiting enzyme for β-oxidation of long-chain fatty acids, carnitine palmitoyl transferase-1, was up-regulated in the liver of fish fed the high-arginine diet. Taken together, the data support that increased dietary arginine activates polyamine turnover and β-oxidation in the liver of juvenile Atlantic salmon and may act to improve the metabolic status of the fish.


[Gerontological aspects of improving the quality of medical and social assistance to older displaced persons in the context of the COVID-19 pandemic in Ukraine.]
This work aimed to study the organization of medical-social services for the elderly internally displaced persons at the level of primary medico-sanitary aid (PMSA) in Ukraine, assessment of the main tasks and specifics of organization and provision of palliative and hospice care (PHC) under COVID-19 conditions. As a result of the sociological investigation, using the questionnaire methods and experts' evaluations, we have analyzed the socio-psychological characteristics of the elderly internally displaced persons, the specifics of their social adaptation, financing and medico-social servicing at the level PMSA, including structural modernization and optimization of organizational provision. The introduction of an affordable and economically viable system allows for the interaction of specialists from health care and social protection institutions, and optimizes compliance with the standards and principles of medical care. Within the framework of the PНC, we use the European approaches of formation of the strategy of palliative care expansion under COVID-19 pandemic. One of the main tasks is the provision of constant PHC. Introduction of the organization-structural system of the medical-social care at the PMSA level and provision of accessible and highly qualitative constant PHC are actual for the elderly displaced people, who are in the group of high risk COVID-19 death group.


Thyroid ultrasonography B-mode and color-Doppler.
Recently, ultrasonography of the thyroid gland has proved to be a useful clinical diagnostic method. The newly developed high-resolution ultrasonography with color-Doppler flow mapping function can reveal fine details of the thyroid gland and the hemodynamic features of thyroid neoplasmas. Ultrasonography can deliver a diagnostic accuracy of over 90% in thyroid carcinoma, especially papillary carcinoma. However, in the case of follicular carcinoma, neither conventional B-mode ultrasound imaging nor aspiration biopsy cytology can deliver satisfactory results. Color-Doppler imaging examination of 212 follicular tumors has been carried out, resulting in the characteristic finding that a high-velocity pulsative blood flow penetrates the tumor in the case of follicular carcinoma, which led to new diagnostic criteria with color-Doppler examination. The differential diagnostic grading score using color-Doppler examination and FFT analysis can show a dramatic diagnostic accuracy of 81.0%, a sensitivity of 88.9% and a specificity of 74.2%. Ultrasound with the color-Doppler function can play a more important role in the differential diagnosis of thyroid tumors.


Development of mouse embryos following the destruction of blastomeres.
Preimplantation mouse embryos in different stages of development were cultured in vitro following the puncture of one or more blastomeres through the zona pellucida. The percentage of embryos that developed to the blastocyst stage was lower when puncturing was done in later stages than in earlier stages, thus showing that regulation capability diminishes as development proceeds. Cell debris of punctured blastomeres were always found under the zona pellucida, in any sector of the blastocyst, and did not prevent the sealing between the underlying cells. The plasma membrane of the punctured blastomeres vacuolized and disappeared within minutes of the operation. The space left by punctured external blastomeres of 16-cell morulae was filled by neighbour peripheral cells and not by inner cells, suggesting cell differentiation prior to blastocoel formation.


3-Hydroxy-3-methylglutaryl--coenzyme A reductase inhibitors in the treatment of hypercholesterolemia.
A new class of drugs, which inhibit de novo cholesterol biosynthesis, significantly reduces the blood cholesterol concentrations in hypercholesterolemic patients. Four separate inhibitors have lowered plasma total cholesterol and low-density lipoprotein (LDL) levels in humans by 20% to 40%: mevastatin (Compactin), lovastatin (mevinolin), pravastatin (CS-514, Eptastatin, and SQ 31000), and simvastatin (Synvinolin, MK-733). In addition to lowering total and LDL cholesterol concentrations, the plasma concentration of the potentially atherogenic B apolipoprotein is also reduced by 20% to 40%. The reduction in the levels of circulating atherogenic lipoprotein particles occurs as a result of decreased synthesis and enhanced removal of LDLs by the LDL receptor pathway in hepatocytes. Moreover, the levels of high-density lipoprotein cholesterol, which are inversely related to atherosclerosis, increase in concentration with treatment by these drugs. If the short-term safety of these drugs extends to ongoing long-term studies and if cardiovascular morbidity and mortality are affected by their use, this class of hypolipidemic agent will markedly facilitate the effective treatment of hypercholesterolemia.


Long-term results of coronary artery bypass surgery in patients with severely depressed left ventricular function.
The objective of the present study was to evaluate medium- and long-term results of coronary artery bypass grafting (CABG) in patients with severe left ventricular dysfunction (LVD). Prospective evaluation (clinical follow-up and equilibrium radionuclide angiography scan) of all the patients with severe LVD who underwent CABG from November 1986 to November 1991 at the Tel Aviv Medical Center and were referred to the Post Cardiac Surgery Follow-up Clinic at this institution. Seventy-four consecutive patients (65 men, 9 women, aged 43 to 82 years; mean age, 68.2 years) with left ventricular ejection fraction (LVEF) of 30% or less who underwent isolated CABG (without automatic implantable cardioverter-defibrillator implantation, aneurysmectomy, valve replacement, or other open heart procedures) during a 5-year period and were discharged from hospitalization were prospectively evaluated. Preoperatively, 62% of patients had angina, 65% had congestive heart failure (CHF), and the LVEF ranged from 10 to 30%. The mean number of grafts was 2.98 per patient; the internal mammary artery (IMA) was used in 54 patients. The patients were followed up 4 to 96 months (mean, 64.9 months) post-surgery for survival, clinical status, and left ventricular function. Survival was 96%, 93.2%, 91.9%, 87.8%, 86.5%, 83.8%, and 83.8%, at 1, 2, 3, 4, 5, 6, and 7 years, respectively. Postoperatively, mean angina class improved from 2.9 to 1.4 (p < 0.0001) and mean CHF class improved from 2.7 to 1.8 (p < 0.0001). Mean LVEF improved from 23.5% preoperatively to 35.7% postoperatively (p < 0.0001). The following occur in patients with coronary artery disease and severe LVD undergoing CABG: (1) good medium- and long-term survival is attained; (2) angina class improves; (3) CHF class improves; (4) LVEF objectively improves; and (5). IMA can be used safely as a conduit.


The relative importance of modifiable potential risk factors of type 2 diabetes: a meta-analysis of two cohorts.
Lifestyle factors predict type 2 diabetes occurrence, but their effect in high- and low-risk populations is poorly known. This study determines the prediction of low-risk lifestyle on type 2 diabetes in those with and without metabolic syndrome in a pooled sample of two representative Finnish cohorts, collected in 1978-1980 and 2000-2001. Altogether 8,627 individuals, aged 40-79 years, and free of diabetes and cardiovascular disease at baseline were included in this study. A low-risk lifestyle was defined based on body mass index, exercise, alcohol consumption, smoking, and serum vitamin D concentration. The metabolic syndrome was defined according to the International Diabetes Federation including obesity, blood pressure, serum HDL cholesterol, serum triglycerides, and fasting glucose. During a 10-year follow-up, altogether 226 type 2 diabetes cases occurred. Overweight was the strongest predictor of type 2 diabetes (population attributable fraction (PAF) = 77%, 95% confidence interval (CI): 53, 88%). Together with lack of exercise, unsatisfactory alcohol consumption, smoking, and low vitamin D concentration it explained 82% of the cases. Altogether 62% (CI: 47, 73%) of the cases were attributable to the metabolic syndrome and 92% (CI: 67, 98%) to the most unfavourable combination of its components. The metabolic syndrome did not modify the prediction of lifestyle factors but persons with normal blood pressure benefited more from positive changes in exercise, alcohol consumption, and smoking than those with elevated blood pressure (P for interaction = 0.01). In conclusion, modification of lifestyle factors apparently reduces type 2 diabetes risk, especially in persons with normal blood pressure.


Novel predictors of intrapulmonary vascular dilatations in cirrhosis: extending the role of pulse oximetry and echocardiography.
Intrapulmonary vascular dilatations (IPVDs) are a criterion for the diagnosis of hepatopulmonary syndrome in patients with liver cirrhosis. We aimed to show that IPVDs are more common than suspected in a heterogenous cirrhotic population and to identify new diagnostic parameters. Forty-three consecutive patients with cirrhosis admitted to our Gastroenterology department were included in this prospective study. History, physical examination, ECG and, when warranted, pulmonary function tests and chest radiograph were used to exclude patients with significant cardiac or pulmonary disease. Contrast enhanced transthoracic echocardiography (CEE) was used to determine the presence of IPVDs. Pulse oximetry readings were taken in the supine and standing positions. We found 12 patients with IPVDs. Statistical analysis proved the correlation between IPVDs and systolic pulmonary artery pressure (sPAP) (p= .049), right ventricle wall width (RVW) (p = .013) and E/A ratio (p = .034) but not left atrial or ventricular diameter. Orthodeoxia was also present more frequently in patients with positive CEE. The difference between supine and standing oxygen saturation (changeSat) proved a fair diagnostic test for detecting IPVDs, with an area under the receiver operated curve (AUROC) of 0.823. Our study shows that RVW, sPAP, E/A and orthodeoxia determined by pulse oximetry are valuable novel predictors of IPVDs, encouraging the routine use of pulse oximetry and echocardiography in cirrhotic patients.


Iron fortification of wheat flour: bioavailability studies.
Bioavailability refers to that fraction of nutrients which is utilized by the body out of the total indigested amount. Various direct and indirect methods for the determination of bioavailability are available. We determined the bioavailability of iron from fortified wheat flour using both in vitro and in vivo methods. The bioavailability data will be used to make the recommendation for a fortification strategy in Pakistan. The in vitro bioavailability of iron from fortified wheat flour was determined using in vitro enzymatic digestion and fermentation by simulating the condition of the small intestine and colon in the laboratory. Different products were prepared from the fortified ferrous sulfate (FeSO4) and unfortified wheat flour. The total iron of the samples was measured by the wet-digestion method and analyzed on an atomic absorption spectrometer (AAS). To obtain the percentage of iron released, the samples were subjected to pepsin digestion and dialysis. The dialysate was collected at 3, 6, 9, and 12 hours and read on an AAS. The retentates from the above were subjected to the fermentation condition of the colon by inoculating it with human fecal inoculum and incubating it for 24 hours at 37 degrees C under anerobic conditions. The dialysate was collected at 3- and 6-hour intervals and read on an AAS. More iron was released from fortified wheat flour (4.6%), leavened chapati (6.8%), and Nan (15.1%) than from the unfortified control samples. Fermentation and leavening resulted in a better release, which was evident from in vitro digestion results.


Hybrid Polymer-Network Hydrogels with Tunable Mechanical Response.
Hybrid polymer-network gels built by both physical and covalent polymer crosslinking combine the advantages of both these crosslinking types: they exhibit high mechanical strength along with excellent fracture toughness and extensibility. If these materials are extensively deformed, their physical crosslinks can break such that strain energy is dissipated and irreversible fracturing is restricted to high strain only. This mechanism of energy dissipation is determined by the kinetics and thermodynamics of the physical crosslinking contribution. In this paper, we present a poly(ethylene glycol) (PEG) based material toolkit to control these contributions in a rational and custom fashion. We form well-defined covalent polymer-network gels with regularly distributed additional supramolecular mechanical fuse links, whose strength of connectivity can be tuned without affecting the primary polymer-network composition. This is possible because the supramolecular fuse links are based on terpyridine⁻metal complexation, such that the mere choice of the fuse-linking metal ion adjusts their kinetics and thermodynamics of complexation⁻decomplexation, which directly affects the mechanical properties of the hybrid gels. We use oscillatory shear rheology to demonstrate this rational control and enhancement of the mechanical properties of the hybrid gels. In addition, static light scattering reveals their highly regular and well-defined polymer-network structures. As a result of both, the present approach provides an easy and reliable concept for preparing hybrid polymer-network gels with rationally designed properties.


F-actin forms transient perinuclear shells at the mitosis-interphase transition.
Intermediate filaments and microtubules are known to be involved in establishing and maintaining nuclear shape. F-actin may also be involved in determining nuclear shape, since we have found it associated with reforming nuclei very briefly after cell division. We stained cells from vertebrate tissue cultures (3T3 and NRK-49F) and epidermal cells from an insect with rhodamine-phalloidin and Hoechst #33342 to localize F-actin in relation to the nucleus. We found that F-actin forms shells only around nuclei during reorganization in late mitosis and early interphase. We suggest that perinuclear F-actin shells may be generally present in eukaryotes, but that they are easily missed because of their delicacy and transience.


A comparative study of antimicrobial, anti-quorum sensing, anti-biofilm, anti-swarming, and antioxidant activities in flower extracts of pecan (Carya illinoinensis) and chestnut (Castanea sativa).
Antibiotic resistance, which has increased rapidly in recent years because of uncontrolled and unconscious antibiotic consumption, poses a major threat to public health. The inadequacy of existing antibiotics has increased the need for new, effective, and less toxic antibiotic raw materials or antibiotic derivatives. Pecan (Carya illinoinensis) and Chestnut (Castanea sativa) flowers possess abundant pollen contents and exhibit similar morphological features. The purpose of this study was to compare these two flower extracts in terms of their antimicrobial and antioxidant activities. Total phenolic content, total flavonoid contents, and phenolic components were also analyzed in aquatic and ethanolic extracts. Antioxidant activities were measured using ferric reducing/antioxidant capacity (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods. Antimicrobial and antifungal activities were compared by means of agar diffusion tests against bacteria including Staphylococcus aureus, Bacillus cereus, Mycobacterium smegmatis, Acinetobacter haemolyticus, and Chromobacterium violaceum, and the yeasts Candida albicans and Candida parapsilosis. Anti-quorum sensing (anti-QS), anti-biofilm, and anti-swarming (SW) activities were also studied against Chromobacterium violaceum ATCC 31532, Chromobacterium violaceum ATCC 12472, and Pseudomonas aeruginosa PA01, respectively. Both extracts were rich in ellagic acid and gallic acid and exhibited similar antioxidant properties. Both flower extracts exhibited high antimicrobial and antifungal activities as well as anti-biofilm, anti-QS, and anti-SW activities.


Psychiatric aspects of heart transplantation.
There are many practical psychiatric, social, and ethical problems which accompany heart transplantation. These include pre- and postoperative anxiety and depressive conditions, post-operative delirium, and social and family dysfunctional syndromes. This paper reviews the literature critically in the following five areas: pre-transplant evaluation, coping with surgery, postoperative sequelae, rehabilitation, and management. Although most recipients have a good outcome from the physical and psychiatric points of view, a substantial minority experience family conflicts and sexual dysfunction after surgery. An average of 45% of patients from all studies returned to full-time employment. The distinctive role and contribution of the psychiatrist on the transplant team is discussed and important areas for future research are outlined.


Correlation of viral RNA biosynthesis with glucose-6-phosphate dehydrogenase activity and host resistance.
Changes in glucose-6-phosphate dehydrogenase (G6P DH; EC 126.96.36.199) activity caused by infection of tobacco ( Nicotiana tabacum L.) leaves with potato virus Y (PVY), cucumber mosaic virus, potato virus X, tobacco rattle virus and turnip mosaic virus, the subcellular localisation of G6P DH isozymes in mesophyll protoplasts derived from healthy and PVY-infected tobacco leaves, as well as G6P DH control and the relationship of its isozymes with the degree of tobacco resistance to PVY multiplication, were studied. The activities of G6P DH were markedly increased in locally and systemically infected leaves and the time courses of the activity linearly correlated with those of virus multiplication. In leaves infected with PVY, the activity time courses of the crude and the partially purified G6P DH were coincident. This probably indicates the involvement of coarse regulation of the enzyme. PVY content linearly correlated with enhanced G6P DH activity in leaf discs derived from susceptible, tolerant and resistant cultivars of tobacco. The increased activity of the enzyme in infected protoplasts and plant tissues was predominantly caused by the increased activity of chloroplastic isozymes. This was confirmed by the specific staining of isozymes after electrophoretic separation of chloroplastic proteins of tobacco leaves. These findings enable the degree of resistance to virus multiplication to be quantified for the use of gene manipulation and breeding.


A correlation of phylogenetic diversity in the Proteobacteria with the influences of ecological forces.
The Proteobacteria are physiologically and morphologically diverse, although they form a coherent set of four main lineages on phylogenetic analysis of ribosomal RNA. A rational and consistent taxonomic arrangement bringing today's phenotypic and phylogenetic conclusions about them into register is not yet possible. It is also difficult to understand the selective forces involved in their evolution that fostered such diversity. This latter problem is addressed in this essay and is based on the assumption that bacterial evolution could only have occurred in ecological consortia whose products of metabolism modified the environment, provided nutrition, and have a basis for selection of new capabilities.


Spindle Cell Melanoma Presenting as an Ulcer in a Black Diabetic.
Background. Melanoma in blacks is uncommon and exceedingly rare in association with a diabetic ulcer. We present a case of a spindle cell melanoma masquerading as a diabetic ulcer. Case Report. A 57-year-old overweight woman presented to The Maria Holder Diabetes Centre for the Caribbean with a nonhealing ulcer of the right heel after being treated by various primary care physicians over the preceding year. Her general and systematic examinations were unremarkable. There was a 1 × 1.5 cm ulcer with a necrotic base which bled easily on contact with no evidence of peripheral neuropathy nor arterial insufficiency. Microscopic examination of a biopsy of the lesion showed fascicles of spindle cells with plump nuclei and intracytoplasmic yellow-brown pigment. Immunohistochemistry confirmed a diagnosis of melanoma. Discussion. There should be a high index of suspicion of malignancy with nonhealing diabetic ulcer especially when coupled with short disease duration. This case highlights the importance of a biopsy and histological evaluation in ulcers presenting in recently diagnosed diabetics with no evidence of peripheral neuropathy or vascular disease. Melanoma should be considered in spindle cell lesions especially with pigment and residual nevus cells.


[Treating depression? All secrets exposed! The tricks of the trade!].
Depression is highly prevalent in the general population and is also costly to society. Although a wide range of pharmacological and psychological antidepressant therapies is available, the vast majority of people with depression remain unrecognised and untreated. Most depressed patients will respond to appropriate treatment, but the clinical management of depression involves far more than antidepressant drugs alone. Treatment failure, or refractory depression, can be overcome when therapeutic strategies adopt a logical, comprehensive and long-term approach. Understanding the interplay of clinical and neurochemical factors associated with the pathogenesis of depression can help physicians to find effective therapeutic alternatives for most of the depressed patients under their care.


Effect of unfractionated heparin, enoxaparin and sulodexide on the relations between secretion and expression of OPG, RANKL and vWF in HUVEC.
Heparin modulates function of vascular endothelium. We studied the effects of unfractionated heparin (UFH) vs. enoxaparin vs. sulodexide on the levels and gene expression of osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kB Ligand (RANKL) and von Willebrand factor (vWF) in Human Umbilical Vein Endothelial Cells (HUVEC) culture. HUVEC were isolated from human umbilical vein by a standard method. The supernatant concentrations (ELISA) and gene expression (Real Time-PCR) of OPG, RANKL and vWF in HUVEC were determined after incubation with various concentrations of UFH, enoxaparin and sulodexide for up to 16 hours. In control HUVEC strong positive correlation between OPG and vWF levels was observed, whereas sRANKL negatively correlated with OPG and vWF levels. Only in control HUVEC a negative correlation between the supernatant level of vWF and its gene expression was found. Already the lowest concentration of UFH caused 2.5-fold increase in OPG gene expression while higher UFH concentrations substantially increased RANKL mRNA level. A negative correlation between the OPG and sRANKL concentration was noticed in supernatant HUVEC which were incubated with enoxaparine. In conclusion, the observed interrelationships between OPG, RANKL and vWF levels in unstimulated HUVEC support the presumption of the pathophysiological links between these proteins. Of the tested heparin formulas UFH seems to be the most potent in altering the OPG, RANKL and vWF axis.


Multi-Dimensional Social Relations and Mental Health among Rural Older Adults.
To date, knowledge of the association between social-relational patterns and health among rural elders in non-Western social and cultural contexts is scant. Using regional population data in rural South Korea, this study examines comprehensive social relations among rural elders, and the extent to which heterogeneous patterns of social relations were associated with cognitive impairment and depression. Using comprehensive social-relational indicators with (a) the structure of social relations (b) the quality of social relations, we applied latent class analysis identified four subgroups of social relations among rural older adults: Restricted, Spouse Focused-Ambivalent, Community Engaged-Positive, and Diverse. We found that the Community-Engaged group was less likely to be depressed as well as have co-ocurring problems of cognitive impairment and depression.


Case study: factors in defining the nurse informatics specialist role.
Healthcare organizations, consultant groups, vendor companies, and academic institutions feel the challenge to enhance user experiences with information systems. To meet this challenge, organizations and companies are looking to better understand and utilize a variety of informatics roles to further marketing, business, or healthcare goals. Nursing is one practice area that can support the successful integration of information systems development, implementation, support, and user experience. However, the definition and development of such a role or position has met with mixed success. This article explores some of the issues and influences related to the role's development. The issues, impacts, and influences have been identified based on healthcare business assessment, job description analysis, employment and project evaluations, and professional standards set by the American Nurses Association.


A protein from the salivary glands of the giant Amazon leech with high sequence homology to a nicotinic acetylcholine receptor subunit.
A gene coding for a soluble protein with homology to the beta subunit of the nicotinic acetylcholine receptor from goldfish was isolated from a cDNA library of Haementeria ghilianii salivary glands. Comparison of the leech protein sequence with the database showed that the N terminus has high homology with the extracellular portion of acetylcholine receptor beta subunits, whilst the C terminus, highly charged, has homology to proteins which may be involved in chelating divalent cations. The leech protein was expressed in mammalian cells and the product compared to the native protein. Both proteins are glycosylated and form polymers which are disrupted by heat but not by reducing agents. A role for this protein in salivary gland secretion is suggested.


Neuroinflammation as a Therapeutic Target in Retinitis Pigmentosa and Quercetin as Its Potential Modulator.
The retina is a multilayer neuronal tissue located in the back of the eye that transduces the environmental light into a neural impulse. Many eye diseases caused by endogenous or exogenous harm lead to retina degeneration with neuroinflammation being a major hallmark of these pathologies. One of the most prevalent retinopathies is retinitis pigmentosa (RP), a clinically and genetically heterogeneous hereditary disorder that causes a decline in vision and eventually blindness. Most RP cases are related to mutations in the rod visual receptor, rhodopsin. The mutant protein triggers inflammatory reactions resulting in the activation of microglia to clear degenerating photoreceptor cells. However, sustained insult caused by the abnormal genetic background exacerbates the inflammatory response and increases oxidative stress in the retina, leading to a decline in rod photoreceptors followed by cone photoreceptors. Thus, inhibition of inflammation in RP has received attention and has been explored as a potential therapeutic strategy. However, pharmacological modulation of the retinal inflammatory response in combination with rhodopsin small molecule chaperones would likely be a more advantageous therapeutic approach to combat RP. Flavonoids, which exhibit antioxidant and anti-inflammatory properties, and modulate the stability and folding of rod opsin, could be a valid option in developing treatment strategies against RP.


The patient satisfaction with nursing care quality: the psychometric study of the Serbian version of PSNCQ questionnaire.
Patient satisfaction with nursing is the most important predictor of patients' overall satisfaction with their hospital care. According to the Law of Health Care of Republic of Serbia monitoring of patients' satisfaction with hospital service is mandatory; however, the questionnaire applied to that purpose includes only several questions directly addressing the nursing care issue. The aim of the present study was to evaluate psychometric properties of the Serbian version Patient Satisfaction Nursing Care Quality Questionnaire (PSNCQQ) and explore patients' satisfaction of nursing care they received and assess the relationship between patient satisfaction and patient characteristics. This cross-sectional study included a sample population of 240 patients who were discharged from surgical clinics of the Clinical Center of Vojvodina in Novi Sad. The PSNCQQ was translated into Serbian according to standard procedures for forward and backward translation. Factor analysis was used to determine the construct validity, and predictive validity of the questionnaire was previously assessed. Cronbach's α coefficient and item analysis was conducted to evaluate reliability of the scale. The Serbian version Patient Satisfaction Nursing Care Quality Questionnaire (PSNCQQ) showed a one-factor structure, Cronbach's α reliability coefficient was excellent 0.94 and was similar across hospital categories. The correlation coefficient between 19 items and the total scale was high, and ranged from 0.56 to 0.76. Patients' age, educational level and previous hospitalization period were important factors that affected their satisfaction with nursing care. The study provides a new tool for measuring patient satisfaction with nursing care in Serbia that may present a useful instrument for nursing care managers in improving the nursing care process.


MetaClean: a machine learning-based classifier for reduced false positive peak detection in untargeted LC-MS metabolomics data.
Despite the availability of several pre-processing software, poor peak integration remains a prevalent problem in untargeted metabolomics data generated using liquid chromatography high-resolution mass spectrometry (LC-MS). As a result, the output of these pre-processing software may retain incorrectly calculated metabolite abundances that can perpetuate in downstream analyses. To address this problem, we propose a computational methodology that combines machine learning and peak quality metrics to filter out low quality peaks. Specifically, we comprehensively and systematically compared the performance of 24 different classifiers generated by combining eight classification algorithms and three sets of peak quality metrics on the task of distinguishing reliably integrated peaks from poorly integrated ones. These classifiers were compared to using a residual standard deviation (RSD) cut-off in pooled quality-control (QC) samples, which aims to remove peaks with analytical error. The best performing classifier was found to be a combination of the AdaBoost algorithm and a set of 11 peak quality metrics previously explored in untargeted metabolomics and proteomics studies. As a complementary approach, applying our framework to peaks retained after filtering by 30% RSD across pooled QC samples was able to further distinguish poorly integrated peaks that were not removed from filtering alone. An R implementation of these classifiers and the overall computational approach is available as the MetaClean package at https://CRAN.R-project.org/package=MetaClean . Our work represents an important step forward in developing an automated tool for filtering out unreliable peak integrations in untargeted LC-MS metabolomics data.


Will health fund rationalisation lead to significant premium reductions?
It has been suggested that rationalisation of health funds will generate significant albeit unquantified cost savings and thus hold or reduce health fund premiums. 2001-2 Private Health Industry Administration Council (PHIAC) data has been used to analyse these suggestions. Payments by funds for clinical services will not vary after fund rationalisation. The savings after rationalisation will arise from reductions in management expenses, which form 10.9% of total fund expenditure. A number of rationalisation scenarios are considered. The highest theoretical industry wide saving found in any plausible scenario is 2.5%, and it is uncertain whether this level of saving could be achieved in practice. If a one off saving of this order were achieved, it would have no medium and long term impact on fund premiums increases given funds are facing cost increases of 4% to 5% per annum due to demographic changes and age standardised utilization increases. It is suggested discussions on fund amalgamation divert attention from the major factors increasing fund costs, which are substantially beyond fund control.


Properties and regulation of pH-dependent cation channels in the apical membrane of cultured proximal tubule cells.
The established opossum kidney (OK) cell line serves as a model system for ion and substrate transport in the renal proximal tubule. Previous experiments on OK cells revealed a channel-mediated Na+ conductance which is regulated by intracellular pH (pHi). In this study we report on patch clamp experiments determining the properties and pHi dependence of a cation channel located in the apical membrane. This channel is selective for sodium over chloride but discriminates poorly between the monovalent cations Na+,K+,Li+ and Cs+. Its open probability (P(o)) rises at depolarising membrane potentials. Under normal conditions the channel is inactive in the cell-attached patch mode and is activated upon excision. However, after excision the channel usually runs down within 30-90 s which cannot be overcome by either altering the Ca(2+)-concentration (10(-3) mol/l, 10(-6) mol/l, Ca(2+)-free) or adding 1 mmol/l Mg-ATP to the bath solution. In the cell-attached patch mode the channel could be activated by decreasing pHi from pH 7.4 to pH 6.5, by either the ammonium prepulse technique or the nigericin K+ method, in more than 50% of the experiments performed. In the renal proximal tubule such a non-selective cation channel would constitute a functional Na+ channel and might therefore support Na+ reabsorption especially during the intracellular acidification due to hormonal inhibition of the Na+/H+ exchanger.


Cytokine plasma levels: reliable predictors for radiation pneumonitis?
Radiotherapy (RT) is the primary treatment modality for inoperable, locally advanced non-small-cell lung cancer (NSCLC), but even with highly conformal treatment planning, radiation pneumonitis (RP) remains the most serious, dose-limiting complication. Previous clinical reports proposed that cytokine plasma levels measured during RT allow to estimate the individual risk of patients to develop RP. The identification of such cytokine risk profiles would facilitate tailoring radiotherapy to maximize treatment efficacy and to minimize radiation toxicity. However, cytokines are produced not only in normal lung tissue after irradiation, but are also over-expressed in tumour cells of NSCLC specimens. This tumour-derived cytokine production may influence circulating plasma levels in NSCLC patients. The aim of the present study was to investigate the prognostic value of TNF-alpha, IL-1beta, IL-6 and TGF-beta1 plasma levels to predict radiation pneumonitis and to evaluate the impact of tumour-derived cytokine production on circulating plasma levels in patients irradiated for NSCLC. In 52 NSCLC patients (stage I-III) cytokine plasma levels were investigated by ELISA before and weekly during RT, during follow-up (1/3/6/9 months after RT), and at the onset of RP. Tumour biopsies were immunohistochemically stained for IL-6 and TGF-beta1, and immunoreactivity was quantified (grade 1-4). RP was evaluated according to LENT-SOMA scale. Tumour response was assessed according to RECIST criteria by chest-CT during follow-up. In our clinical study 21 out of 52 patients developed RP (grade I/II/III/IV: 11/3/6/1 patients). Unexpectedly, cytokine plasma levels measured before and during RT did not correlate with RP incidence. In most patients IL-6 and TGF-beta1 plasma levels were already elevated before RT and correlated significantly with the IL-6 and TGF-beta1 production in corresponding tumour biopsies. Moreover, IL-6 and TGF-beta1 plasma levels measured during follow-up were significantly associated with the individual tumour responses of these patients. The results of this study did not confirm that cytokine plasma levels, neither their absolute nor any relative values, may identify patients at risk for RP. In contrast, the clear correlations of IL-6 and TGF-beta1 plasma levels with the cytokine production in corresponding tumour biopsies and with the individual tumour responses suggest that the tumour is the major source of circulating cytokines in patients receiving RT for advanced NSCLC.


Incidence, Reversal, and Prevention of Opioid-induced Respiratory Depression.
Opioid treatment of pain is generally safe with 0.5% or less events from respiratory depression. However, fatalities are regularly reported. The only treatment currently available to reverse opioid respiratory depression is by naloxone infusion. The efficacy of naloxone depends on its own pharmacological characteristics and on those (including receptor kinetics) of the opioid that needs reversal. Short elimination of naloxone and biophase equilibration half-lives and rapid receptor kinetics complicates reversal of high-affinity opioids. An opioid with high receptor affinity will require greater naloxone concentrations and/or a continuous infusion before reversal sets in compared with an opioid with lower receptor affinity. The clinical approach to severe opioid-induced respiratory depression is to titrate naloxone to effect and continue treatment by continuous infusion until chances for renarcotization have diminished. New approaches to prevent opioid respiratory depression without affecting analgesia have led to the experimental application of serotinine agonists, ampakines, and the antibiotic minocycline.


Robotic thymectomy-a new approach for thymus.
Advancements in modern technology bring many evolutions in minimally invasive surgery such as robot assisted approaches. Because of complete resection is so important in thymectomy operations, they became a new era for robotic surgery as a result of its superiorities (intuitive movements, tremor filtration, more degrees of manipulative freedom, motion scaling, and high-definition stereoscopic vision).


Frequency of bronchopneumonia in children with survival interval before death.
Many children do not survive after presentation in extremis. Some survive varying intervals and are found to have bronchopneumonia at death. The question is raised whether bronchopneumonia is a consequence of survival rather than the initiating disease leading to collapse. A prospective study of the deaths of 156 children divided them into two groups: 80 children with head injury and 76 with causes of death other than sudden infant death syndrome. In 43 of the total group of children, bronchopneumonia was found. In the total group, 76 survived more than a day. Of these 39 had bronchopneumonia, 32 died of head injury, and 7 had other causes of death. Of the children surviving less than a day, 4 had bronchopneumonia at death--only 1 with head injury. If bronchopneumonia is present, it is more likely to have developed after the collapse than to have caused it in this population.


Brugada syndrome unmasked by fever: a comprehensive review of literature.
The Brugada pattern is identified on the EKG by a coved ST-segment elevation accompanied by a negative T wave in the early precordial leads in the absence of a cardiac structural abnormality. Brugada pattern and Brugada syndrome should be differentiated, as the latter is associated with an increased risk of sudden cardiac death. The literature was searched using multiple databases to identify all the articles on Brugada pattern. Data were screened and analyzed by independent authors. Sixty articles, comprising 71 patients, were included in the study. The mean age of patients was 42.6 years, with a higher prevalence of Brugada pattern in men (83%) than women (17%). The most frequent findings associated with Brugada pattern was fever (83%). Other less common presentations included cough (21%), sore throat (10%), syncope (18%), abdominal pain (8%), and chest pain (7%). Comorbidities included pneumonia (30%), upper respiratory tract infections (14%) and smoking (14%). Among treatment modalities, 39% of patients had ICD placement performed, 44% received antibiotics, while 14% had supportive care. Adenosine was given to 3% of patients, while other antiarrhythmics like milrinone, amiodarone, sotalol, procainamide, flecainide, and nitroglycerin were given to 1% of patients. Most patients with Brugada syndrome had a satisfactory outcome, with only 4% mortality rate(WHAT ABOUT THE OTHER 11%?). Out of the 71 patients, 3% had persistent Brugada patterns, while 86% of patients recovered completely. There was no significant effect of ICD on mortality or Brugada pattern resolution (p 0.37). Our study shows that fever is the main reason for unmasking the Brugada pattern in patients with this channelopathy. ICD placement in such patients is not recommended as it has no mortality benefits.


The effects of time of disease occurrence, milk yield, and body condition on fertility of dairy cows.
The associations between occurrence of diseases, milk yield, and body condition score on conception risk after first artificial insemination (AI) were analyzed in an observational study on a convenience sample of 43 farms participating in a herd health program. Data were taken from 9369 lactations, from 4382 cows inseminated between 20 and 180 d in milk from 1990 to 1996. Two logistic regression models, one containing data from all lactations and a subset containing data from 1762 lactations with body condition scoring, were used to determine pregnancy risk at first AI. The effects of herd deviation in test-day milk yield, body condition score loss, and milk fat to protein ratio changes in early lactation were significant predictors of pregnancy risk, independent of disease; days in milk; farm; and seasonal factors. Three different methods of disease parameterization (incidence rates, binomial classes dependent on the interval in days since last occurrence with respect to AI, and a linear variable weighted for this interval) produced similar results. Metritis, cystic ovarian disease, lameness, and mastitis gave odds ratios for pregnancy risk ranging from 0.35 to 1.15, largely dependent on the interval in days from final disease occurrence to first AI. Displaced abomasum, milk fever, and retained fetal membranes resulted in odds ratios for pregnancy risk of 0.25, 0.85, and 0.55, respectively. These diseases showed little relationship between fertility and the number of days since last occurrence. Results of this study confirm the negative effects of milk yield, body score condition loss, and disease on dairy cow fertility. The effects of some diseases on first service conception were strongly dependent on the interval since last disease occurrence. This was especially valid for clinical mastitis, which has an extremely weak effect on conception if occurring prior to AI and is associated with > 50% reduction in pregnancy risk if occurring in the 3 wk directly after AI.