Association between insomnia severity and presenteeism among Japanese daytime workers.
Presenteeism is the loss of productivity due to a worker's health problems, despite the worker being present at the workplace. Although the association between presenteeism and insomnia complaints is well known, few studies have examined the association between insomnia severity and presenteeism. This study aimed to explore the association between insomnia severity and presenteeism. This study included 1925 participants (1543 males, 379 females, and three of other genders) in total. The mean age of the participants was 49.94 ± 9.82 years. The inclusion criteria were full-time employment, working 8 hr per day and 5 days per week, and having no night shifts. The insomnia severity was classified based on the Insomnia Severity Index and Athens Insomnia Scale criteria. Logistic regression analysis showed that moderate and severe insomnia severity were associated to a greater magnitude with presenteeism than mild insomnia severity. Severe insomnia severity was associated to a greater magnitude with presenteeism than moderate insomnia severity. In summary, logistic regression analysis showed that increased insomnia severity based on Insomnia Severity Index classification was associated with increased odds of presenteeism, but increased insomnia severity based on Athens Insomnia Scale classification was not fully consistent with increased odds of presenteeism. Because increased insomnia severity is associated with worsening of presenteeism, early detection of and early intervention against insomnia complaints are important for reducing presenteeism. This study was the first to examine the associations between presenteeism and insomnia severity classification of no insomnia, mild, moderate, and severe insomnia severity.


Graft survival and risk factors of penetrating keratoplasty for microbial keratitis.
To evaluate graft survival rates and prognostic factors in relation to penetrating keratoplasty for microbial keratitis. The records of 95 patients treated with penetrating keratoplasty for microbial keratitis during a twenty-year period were reviewed. Data were analysed by construction of survival curves using the Kaplan-Meier non parametric method. The overall survival of a clear cornea was 72% after one year, 71% after two years and 52% after five years. A significantly lower survival rate (p<0.05) was found in the presence of preoperative local risk factors such as wear of contact lenses and trauma and in inflamed eyes. In contrast, systemic diseases like diabetes, cancer or rheumatoid arthritis did not affect survival and neither did recipient age nor the degree of vascularization of the eye. Male donor buttons showed superior survival compared to female ones (p < 0.05), while females seemed to constitute the best recipients. Recurrence rates of microbial keratitis postoperatively were 11%, 16% and 24% after one, two and five years, respectively. Corresponding graft rejection rates were 9%, 15% and 27%. Patients suffering from microbial keratitis have a relatively high risk of graft failure. To improve the prognosis care should be taken to minimize local risk factors. Surgery should whenever possible be performed on quiet eyes. The unexpected finding of a better prognosis for male donor buttons might suggest the preferred use of male donors in patients suffering from microbial keratitis, but the observation needs further documentation.


Use of cardiovascular procedures among black persons and white persons: a 7-year nationwide study in patients with renal disease.
Black persons historically undergo fewer invasive cardiovascular procedures than white persons. To determine whether acquisition of Medicare health insurance and comprehensive care for severe illness reduce ethnic disparity in use of cardiovascular procedures. 7-year longitudinal analyses in a cohort from the United States Renal Data System. Health care institutions in the United States. Nationwide random sample of 4987 adult black and white patients with incident end-stage renal disease (ESRD) from 303 dialysis facilities in 1986 to 1987. Medical history and service use records, physical examination, and laboratory data. Main outcome measures were receipt of a coronary catheterization or revascularization procedure before (baseline) and after (follow-up) development of ESRD and acquisition of Medicare, adjusted for clinical and socioeconomic variables. At baseline, 9.9% of white patients and 2.8% of black patients had had a cardiac procedure; the odds were almost three times greater in white than in black patients (adjusted odds ratio, 2.92 [95% CI, 2.04 to 4.18]). During follow-up, white patients were only 1.4 times more likely than black patients to have a procedure (adjusted relative risk, 1.41 [CI, 1.13 to 1.77]); rates were 7.8% for white persons and 8.5% for black persons. In patients with Medicare coverage before development of ESRD, the initial three-fold difference in procedure use was eliminated over follow-up (odds ratio, 1.05 [CI, 0.56 to 1.60]). For procedures after hospital admission for myocardial infarction or coronary disease, no difference between ethnic groups was seen during follow-up (relative risk, 1.12 [CI, 0.68 to 1.85]). Differences between ethnic groups in use of cardiovascular procedures narrowed markedly once a serious illness (ESRD) developed and adequate insurance coverage was ensured; the disparity was eliminated in patients with previous Medicare insurance or a stronger indication for a procedure. These findings suggest that almost equal access to care is attainable by combining insurance with delivery of comprehensive, clinically appropriate care.


Tracking the Evolution of Polymer Interface Films during the Process of Thermal Annealing at the Domain and Single Molecular Levels using Scanning Tunneling Microscopy.
Structural evolution of polymer (NTZ12) interface films during the process of annealing is revealed at the domain and single molecular levels using the statistical data measured from scanning tunneling microscopy images and through theoretical calculations. First, common features of the interface films are examined. Then, mean values of surface-occupied ratio, size and density of the domain are used to reveal the intrinsic derivation of the respective stages. Formation of new domains is triggered at 70 °C, but domain ripening is not activated. At 110 °C, the speed of formation of new domains is almost balanced by the consumption due to the ripening process. However, formation of new domains is reduced heavily at 150 °C but restarted at 190 °C. At the single molecular level, the ratio of the average length of linear to curved backbones is increased during annealing, whereas the ratios of the total length and the total number of linear to curved skeletons reaches a peak value at 150 °C. The two major conformations of curved backbones for all samples are 120° and 180° bending, but the ripening at 150 °C reduces 180° folding dramatically. Molecular dynamic simulations disclose the fast relaxing process of curved skeletons at high temperature.


Serological monitoring of paediatric visceral leishmaniasis By IFA and ELISA methods.
Visceral leishmaniasis (VL) is endemic in all Mediterranean countries including Turkey, and children are at greater risk than adults in endemic areas. In VL patients, serological assays are considered to be sensitive for the diagnosis and/or follow up. The aim of this study was to assess the usefulness of serology for following up of childhood VL in Turkey. Sera obtained from twenty parasitologically confirmed children with VL were tested using IFAT and ELISA. The patients were monitored clinically and serologically (range: 20-500 days) during and after treatment. All VL patients were treated with meglumine antimonate. Anti-Leishmania antibodies in successfully treated VL patients showed a steep decline but, in three patients who had relapsed, an increase was detected. Significantly lower values were observed after treatment with both serological techniques. Mean ELISA optical density values before and after treatment were: 0.78 ± 0.36 (0.26-1.76) and 0.38 ± 0.24 (0.09-0.83) respectively, (p < 0.001) and mean IFAT values (log10 transformed titers) before and after treatment were: 3.02 ± 0.90 (1.81-4.51) and 2.16 ± 0.75 (1.20-3.90) respectively, (p < 0.001). ELISA and IFAT are valuable not only for diagnosis but also for monitoring of drug therapy in childhood visceral leishmaniasis as rapid and non-invasive techniques.


Relationship between structural gel and mechanical gel for ABA triblock copolymer in solutions: a molecular dynamics simulation.
Polymer gel exists ubiquitously in our daily life, as in food, cosmetics, drugs, and so on. From the structural point of view, the 3D network can be found in a structural gel. In most experimental work, the gel is identified by the sharp increase in modules; that is, the gel should have similar properties as those of a solid, which is named as mechanical gel. However, not all structural gels have strong mechanical responses. Therefore, studying the relationship between structural gel and mechanical gel is very important. In this work, we investigate the structure and mechanical properties of symmetric ABA copolymers with solvophobic end blocks during the sol-gel transition. Three typical systems with weak, middle, and strong solvophobicities are simulated. It is found that the gelation concentration, gel structure, and mechanical response of structural gel are strongly affected by the solvophobicity of ABA block copolymer. We also find that only the gel formed in strong solvophobic systems has a strong mechanical response. Furthermore, the influence of solvophobicity of A-block on the static and dynamic properties of ABA block copolymers in solutions is also studied to give a molecular understanding of physical gelation.


Quantitative in vitro and in vivo pharmacological profile of CE-178253, a potent and selective cannabinoid type 1 (CB1) receptor antagonist.
Cannabinoid 1 (CB1) receptor antagonists exhibit pharmacological properties favorable for the treatment of obesity and other related metabolic disorders. CE-178253 (1-[7-(2-Chlorophenyl)-8-(4-chlorophenyl)-2-methylpyrazolo[1,5-a]-[1,3,5]triazin-4-yl]-3-ethylaminoazetidine-3-carboxylic acid hydrochloride) is a recently discovered selective centrally-acting CB1 receptor antagonist. Despite a large body of knowledge on cannabinoid receptor antagonists little data exist on the quantitative pharmacology of this therapeutic class of drugs. The purpose of the current studies was to evaluate the quantitative pharmacology and concentration/effect relationships of CE-178253 based on unbound plasma concentration and in vitro pharmacology data in different in vivo preclinical models of FI and energy expenditure. In vitro, CE-178253 exhibits sub-nanomolar potency at human CB1 receptors in both binding (Ki = 0.33 nM) and functional assays (Ki = 0.07 nM). CE-178253 has low affinity (Ki > 10,000 nM) for human CB2 receptors. In vivo, CE-178253 exhibits concentration-dependent anorectic activity in both fast-induced re-feeding and spontaneous nocturnal feeding FI models. As measured by indirect calorimetry, CE-178253 acutely stimulates energy expenditure by greater than 30% in rats and shifts substrate oxidation from carbohydrate to fat as indicated by a decrease the respiratory quotient from 0.85 to 0.75. Determination of the concentration-effect relationships and ex vivo receptor occupancy in efficacy models of energy intake and expenditure suggest that a greater than a 2-fold coverage of the Ki (50-75% receptor occupancy) is required for maximum efficacy. Finally, in two preclinical models of obesity, CE-178253 dose-dependently promotes weight loss in diet-induced obese rats and mice. We have combined quantitative pharmacology and ex vivo CB1 receptor occupancy data to assess concentration/effect relationships in food intake, energy expenditure and weight loss studies. Quantitative pharmacology studies provide a strong a foundation for establishing and improving confidence in mechanism as well as aiding in the progression of compounds from preclinical pharmacology to clinical development.


C2 and C3 pain dermatomes in man.
This report defines the C2 and C3 pain dermatomes by the distribution of: the hypalgesia clearing after surgical root decompression; the dysaesthesias produced by electrical root stimulation; and the hypalgesia produced by anaesthetic root block. The C2 pain dermatome, so defined, consists of an occipital parietal area 6-8 cm wide, ascending paramedially from the subocciput to the vertex. The C3 pain dermatome is a craniofacial area including the scalp around the ear, the pinna, the lateral cheek over the angle of the jaw, the submental region and the lateral and anterior aspects of the upper neck. These C2 and C3 pain dermatomes do not overlap and are smaller than the C2 and C3 tactile dermatomes described in the literature.


Extended Depth of Focus Versus Monofocal IOLs in Patients With High Myopia: Objective and Subjective Visual Outcomes.
To compare the objective and subjective outcomes between the extended depth of focus (EDOF) Mini Well intraocular lens (IOL) and the aspheric monofocal Mini-4-Ready IOL (both SIFI S.p.A.) in patients with high myopia. In this prospective comparative study, 40 patients with high myopia (axial length ≥ 26 mm) were enrolled: 20 patients were bilaterally implanted with the EDOF Mini Well IOL (EDOF group) and 20 patients were bilaterally implanted with the Mini-4-Ready IOL (monofocal group). Three-month follow-up data included corrected and uncorrected distance visual acuity at 4 m and 80, 67, and 40 cm, defocus curves, subjective and objective contrast sensitivity, objective optical quality (calculated with Optical Quality Analysis System; Visiometrics SL), halometry, and reading performance. Subjective visual quality was evaluated with National Eye Institute Refractive Error Quality of Life Instrument 42 scores. All visual acuities were significantly better in the EDOF group (P ⩽ .04) except monocular and binocular uncorrected and corrected distance visual acuities for distance (P ≥ .50). Defocus curves for myopic and hyperopic values were better in the EDOF group (P ⩽ .05), apart from +0.50 to -0.50 D (P ≥ .16). Contrast sensitivity curves was worse in the EDOF group in the mesopic-with-glare condition (P ⩽ .04). No differences were found in halometric values (P ≥ .15) and OQAS outcomes (P ≥ .47). National Eye Institute Refractive Error Quality of Life Instrument 42 subscale scores were better for expectation, near vision, activity limitations, and dependence on correction in the EDOF group (P ⩽ .04). Intermediate and near visual acuities were better in the EDOF group than in the monofocal group, with a comparable visual quality index between groups. [J Refract Surg. 2022;38(3):158-166.].


Thyroid uptake of liquid versus capsule 131I tracers in hyperthyroid patients treated with liquid 131I.
The amount of 131I used to treat hyperthyroid patients is based in part on the 24-hour thyroid uptake of a diagnostic amount of radioiodine (tracer). We compared the 24-hour uptake of an 131I tracer administered in liquid or capsule form to the 24-hour uptake of 131I therapy administered as liquid. Sixty-five hyperthyroid patients with Graves' disease were evaluated and subsequently treated with radioiodine. The liquid group (45 patients) received a liquid 131I tracer (1.85 MBq [0.05 mCi]) and the capsule group (20 patients) received a capsule 131I tracer (1.63 MBq [0.044 mCi]). Probe calibration factors were the same for the liquid and capsule 131I standards. All patients received therapeutic amounts of 131I [114.7-1106.3 MBq [3.1-29.9 mCi]) in liquid form. Therapy uptakes were obtained using the same collimated uptake probe modified with a calibrated lead shield to attenuate the high photon flux. The mean therapeutic uptake was the same for both groups (58%). The mean diagnostic uptake for the capsule group, however, was less than the mean diagnostic uptake for the liquid group (44% vs. 63%). The mean diagnostic uptake for the capsule group was significantly lower than the mean therapeutic uptake for this group (44% vs. 58%), whereas the mean diagnostic and therapeutic uptakes were similar for the group receiving a liquid tracer (63% vs. 58%). In conclusion, diagnostic uptakes performed with a liquid tracer more accurately predicted liquid therapy uptakes than diagnostic uptakes performed with a capsule tracer. This raises concern about the bioavailability of 131I in capsule form and has implications for determining the amount of 131I to administer for therapy. Patients whose 131I therapy was based on the uptake of a capsule tracer received a higher than intended amount of radiation to the thyroid gland.


Impact of water supply and sanitation on diarrheal morbidity among young children in the socioeconomic and cultural context of Rwanda (Africa).
This project studied the frequency and intensity of water contamination at the source, during transportation, and at home to determine the causes of contamination and its impact on the health of children aged 0 to 5 years. The methods used were construction of the infrastructure for three sources of potable water, administration of a questionnaire about socioeconomic status and sanitation behavior, anthropometric measurement of children, and analysis of water and feces. The contamination, first thought to be only a function of rainfall, turned out to be a very complex phenomenon. Water in homes was contaminated (43.4%) with more than 1100 total coliforms/100 ml due to the use of unclean utensils to transport and store water. This socioeconomic and cultural problem should be addressed with health education about sanitation. The latrines (found in 43.8% of families) presented a double-edged problem. The extremely high population density reduced the surface area of land per family, which resulted in a severe nutritional deficit (15% of the children) affecting mainly young children, rendering them more susceptible to diarrhea (three episodes/child/year).


Paroxysmal exercise-induced dystonia within the phenotypic spectrum of ECHS1 deficiency.
ECHS1 encodes a mitochondrial enzyme involved in the degradation of essential amino acids and fatty acids. Recently, ECHS1 mutations were shown to cause a new severe metabolic disorder presenting as Leigh or Leigh-like syndromes. The objective of this study was to describe a family with 2 siblings affected by different dystonic disorders as a resulting phenotype of ECHS1 mutations. Clinical evaluation, MRI imaging, genome-wide linkage, exome sequencing, urine metabolite profiling, and protein expression studies were performed. The first sibling is 17 years old and presents with generalized dystonia and severe bilateral pallidal MRI lesions after 1 episode of infantile subacute metabolic encephalopathy (Leigh-like syndrome). In contrast, the younger sibling (15 years old) only suffers from paroxysmal exercise-induced dystonia and has very mild pallidal MRI abnormalities. Both patients carry compound heterozygous ECHS1 mutations: c.232G>T (predicted protein effect: p.Glu78Ter) and c.518C>T (p.Ala173Val). Linkage analysis, exome sequencing, cosegregation, expression studies, and metabolite profiling support the pathogenicity of these mutations. Expression studies in patients' fibroblasts showed mitochondrial localization and severely reduced levels of ECHS1 protein. Increased urinary S-(2-carboxypropyl)cysteine and N-acetyl-S-(2-carboxypropyl)cysteine levels, proposed metabolic markers of this disorder, were documented in both siblings. Sequencing ECHS1 in 30 unrelated patients with paroxysmal dyskinesias revealed no further mutations. The phenotype associated with ECHS1 mutations might be milder than reported earlier, compatible with prolonged survival, and also includes isolated paroxysmal exercise-induced dystonia. ECHS1 screening should be considered in patients with otherwise unexplained paroxysmal exercise-induced dystonia, in addition to those with Leigh and Leigh-like syndromes. Diet regimens and detoxifying agents represent potential therapeutic strategies. © 2016 International Parkinson and Movement Disorder Society.


Addition of Cocoa Powder, Cane Sugar, and Carrageenan to Milk Enhances Growth of Listeria monocytogenes.
Previously we found that under similar conditions Listeria monocytogenes achieved populations in chocolate milk that were 10 times greater than those in other fluid milk products. The current studies were undertaken to determine why the bacterium grew so well in chocolate milk. Autoclaved samples of milk with 2% milkfat, 2% milk + sugar, 2% milk + cocoa, and 2% milk + sugar + cocoa were inoculated with one of two strains of L. monocytogenes and incubated at 13°C. Carrageenan was also added to one-half of all samples containing cocoa. Growth curves were derived and generation times and maximum populations were calculated for each combination of product and strain of the bacterium. Strain V7 grew faster than strain CA in all products, with most rapid growth occurring in samples containing cocoa (with or without added sugar). Addition of carrageenan further reduced the generation time of this strain. Overall, growth rates ranged from 3 h 55 min (V7 in 2% milk + sugar + cocoa + carrageenan) to 4 h 53 min (CA in 2% milk + cocoa). Product type was primarily responsible for differences in maximum populations achieved by L. monocytogenes . In each instance, final numbers reached were at least 108 cells/ml with highest levels in samples containing all ingredients. The data suggest that sugar, cocoa and carrageenan when added to milk contributed to enhancing growth of L. monocytogenes .


Similar permeability responses to nitric oxide synthase inhibitors of venules from three animal species.
The influence of nitric oxide (NO) on microvascular permeability remains unclear. NO synthase (NOS) inhibitors have been reported to increase as well as to decrease permeability in different experimental models and animal species. We tested the hypothesis that NOS inhibitors influence venular permeability differently in amphibians and mammals. Permeability coefficients to albumin (P(alb)(s)) were measured on in situ mesenteric venules of the frog and rat and excised pig coronary venules before and after exposure to NOS inhibitors. Despite individual variability in magnitude of responses, NOS inhibitors resulted in a reduction in P(alb)(s) in each species. Superfusion with 10(-5) M N(G)-monomethyl-l-arginine (l-NMMA) reduced P(alb)(s) of frog mesenteric venules by 42% (from a median of 11.4 x 10(-7) cm s(-1), n = 12, P < 0.01) and by 67% in porcine coronary venules (from 12.5 x 10(-7) cm s(-1), n = 5, P < 0.05). The response was attenuated in rat mesenteric venules; 10(-4) M N(G)-nitro-l-arginine methyl ester (l-NAME) reduced P(alb)(s) by 23% (from 7.6 x 10(-7) cm s(-1), n = 9, P = 0.01). The inactive d-enantiomers of the NOS inhibitors were without effect on P(alb)(s) in each model. In pig venules, perfusion with blood modified the permeability responses to l-NMMA, suggesting that effects of NO on permeability are modified by one or more elements of blood. These data support a role of nitric oxide release on venular permeability to albumin that is conserved among the three animal species.


Iron-Catalyzed α-Arylation of Deoxybenzoins with Arenes through an Oxidative Dehydrogenative Approach.
A novel α-arylation of deoxybenzoins with non-prefunctionalized arenes is developed through an iron-catalyzed oxidative dehydrogenative approach. The reaction shows broad substrate scope and functional group tolerance and thus provides efficient access to synthetically useful 1,2,2-triarylethanones. A reasonable mechanism is also proposed.


Venous thromboembolic disease: the role of US.
Deep vein thrombosis (DVT) is a ubiquitous process that in the acute setting can lead to pulmonary embolism. Chronically, permanent changes that develop within the venous system following an episode of DVT can produce the postphlebitic syndrome, which is associated with pain, swelling, and ulceration. The postphlebitis syndrome can often mimic acute DVT or coexist with it. The clinical evaluation of DVT is ineffective and necessitates a reliable noninvasive diagnostic technique. Compression ultrasound (US) has proved to be the diagnostic method of choice for detection of extremity clot. Femoral and popliteal veins are routinely evaluated for acute clot, but uncertainty exists concerning the need to evaluate the calf veins similarly. US also can be used to diagnose chronic venous changes, which are indicated by the presence of incompetent valves and retrograde blood flow. Upper-extremity venous thrombosis, often induced by indwelling catheters, can also be diagnosed with US.


Elastin-like polypeptides: A strategic fusion partner for biologics.
Elastin-like peptides (ELPs) are derivatives of tropoelastin with a unique property that allows them to stay soluble below a certain critical temperature but reversibly form aggregates above that temperature. Since they are derived from tropoelastin, ELPs are biocompatible, non-toxic, and non-immunogenic. The unique properties of ELPs have made them a desirable class of fusion tags used in several biomedical applications including targeted drug delivery and enhancing the half-life of protein drugs. The most significant property of an ELP is that when fused to other proteins, the phase transition property of the ELP is maintained, and the target protein can be purified using the thermally driven property of the ELP. The ELP tag purification process is simple and inexpensive, and involves cycling the protein above and below the transition temperature of the ELP fusion followed by centrifugation to obtain the desired protein, without any need for chromatography. Consequently, using ELPs as a purification tag should be potentially interesting to biopharmaceutical companies who spend a significant percentage of their manufacturing costs on expensive protein purification techniques such as chromatography and filtration. However, ELP tags have not yet been used for commercial protein purification due to some challenges of translating this technique, which has been demonstrated mostly in academic laboratories, to a biotechnology manufacturing environment. The article first reviews the state-of-the-art in protein "ELPylation," and discusses some applications which have benefitted from using ELP as a fusion tag. Then, the article discusses the main advantages of using ELP as a purification tag, and evaluates the remaining hurdles for its implementation in industrial protein production. Biotechnol. Bioeng. 2016;113: 1617-1627. © 2016 Wiley Periodicals, Inc.


Efficacy of deep biopsy for subepithelial lesions in the upper gastrointestinal tract.
Accurate diagnosis of subepithelial lesions (SELs) in the gastrointestinal tract depends on a variety of methods: endoscopy, endoscopic ultrasound and different types of biopsy. Making an error-free diagnosis is vital for the subsequent application of an appropriate treatment. To evaluate the efficacy of deep biopsy via the endoscopic submucosal dissection (ESD) technique for SELs in the upper gastrointestinal tract. It was a case series study. Deep biopsy via the ESD technique was completed in 38 patients between November 2012 and October 2014. Thirty-eight SELs in the upper gastrointestinal tract of varying size (very small ≤ 1 cm, small 1-2 cm and large ≥ 2 cm) by means of the ESD technique after an incision with an electrosurgical knife of the overlying layers and revealing a small part of the lesion were biopsied under direct endoscopic view. Deep biopsy via the ESD technique was diagnostic in 28 of 38 patients (73.3%; 95% CI: 59.7-89.7%). The diagnostic yield for SELs with a clear endophytic shape increased to 91.3%. An evident endophytic appearance of a subepithelial lesion, the mean number of biopsied samples (6.65 ±1.36) and the total size in length of all samples per case (19.88 ±8.07 mm) were the main criteria influencing the positiveness of deep biopsy in the diagnostic group compared to the nondiagnostic one (p = 0.001; p = 0.025; p = 0.008). Deep biopsy via the ESD technique is an effective and safe method for the diagnosis of SELs especially with a clear endophytic appearance in a large number of biopsied samples.


Systematic screening for bioactive peptides.
Using simultaneous multiple peptide synthesis by the multipin approach, the feasibility of systematic large-scale pharmacological screening of peptide ligands was investigated. The method involves the assembly of small quantities of peptides (ca. 50 nmol) on plastic pins derivatized with an ester linker based on glycolate and 4-(hydroxymethyl)benzoate. These esters are stable under peptide synthesis and side-chain deprotection conditions but cleave under relatively mild basic conditions to generate peptides having C-terminal acid, amide and methylamide. A two-step approach to side-chain deprotection and cleavage from the solid support allows potentially toxic reagents to be removed (washed) from the peptide prior to cleavage. Consequently, the resulting peptide solutions can be used in bioassays with minimal processing. A series of angiotensin II and substance P analogs were synthesized and evaluated in an in vivo rat model and in vitro radioreceptor assay, respectively. Structure-activity studies on analogs of these bioactive peptides are well documented. The data obtained were consistent with that reported in the literature.


Porcine adenoviruses: an update on genome analysis and vector development.
Although porcine adenoviruses (PAdV) are present in the swine populations worldwide, they usually do not cause any disease, or the infection is only manifested in a mild diarrhoea or respiratory signs. The importance of adenoviruses, however, is constantly growing as there is a possibility of developing them into viral vector vaccines against more significant swine pathogens. A short summary of the well-established facts of porcine adenoviruses is given and recent developments of the genetic analysis of these viruses are discussed in detail. The possibilities of vector development and examples of vector vaccines already reported in the literature are mentioned.


Y chromosome haplotypes in Central-South Italy: implication for reference database.
One hundred and fifty individuals have been sampled across Central-South Italy and genotyped for Y chromosome STRs by PowerPlex Y system. Comparison with previous Italian databases revealed that majority of Y chromosome variation still need to be sampled. Identification of locus duplications, distribution of genetic variation and firstly identified alleles point to the necessity of more focused sampling strategies for reference databases.


Novel peptide linkers for highly potent antibody-auristatin conjugate.
Auristatins are highly potent antimitotic agents that have received considerable attention because of their activities when targeted to tumor cells in the form of antibody-drug conjugates (ADCs). Our lead agent, SGN-35, consists of the cAC10 antibody linked to the N-terminal amino acid of monomethylauristatin E (MMAE) via a valine-citrulline p-aminobenzylcarbamate (val-cit-PABC) linker that is cleaved by intracellular proteases such as cathepsin B. More recently, we developed an auristatin F (AF) derivative monomethylauristatin F (MMAF), which unlike MMAE contains the amino acid phenylalanine at the C-terminal position. Because of the negatively charged C-terminal residue, the potency of AF and MMAF is impaired. However, their ability to kill target cells is greatly enhanced through facilitated cellular uptake by internalizing mAbs. Here, we explore the effects of linker technology on AF-based ADC potency, activity, and tolerability by generating a diverse set of dipeptide linkers between the C-terminal residue and the mAb carrier. The resulting ADCs differed widely in activity, with some having significantly improved therapeutic indices compared to the original mAb-Val-Cit-PABC-MMAF conjugate. The therapeutic index was increased yet further by generating dipeptide-based ADCs utilizing new auristatins with methionine or tryptophan as the C-terminal drug residue. These results demonstrate that manipulation of the C-terminal peptide sequence used to attach auristatins to the mAb carrier can lead to highly potent and specific conjugates with greatly improved therapeutic windows.


Photoelectron spectroscopy and density functional theory study of TiAlO(y) (-) (y=1-3) and TiAl(2)O(y) (-) (y=2-3) clusters.
Small titanium-aluminum oxide clusters, TiAlO(y) (-) (y=1-3) and TiAl(2)O(y) (-) (y=2-3), were studied by using anion photoelectron spectroscopy. The adiabatic detachment energies of TiAlO(y) (-) (y=1-3) were estimated to be 1.11±0.05, 1.70±0.08, and 2.47±0.08eV based on their photoelectron spectra; those of TiAl(2)O(2) (-) and TiAl(2)O(3) (-) were estimated to be 1.17±0.08 and 2.2±0.1eV, respectively. The structures of these clusters were determined by comparison of density functional calculations with the experimental results. The structure of TiAlO(-) is nearly linear with the O atom in the middle. That of TiAlO(2) (-) is a kite-shaped structure. TiAlO(3) (-) has a kite-shaped TiAlO(2) unit with the third O atom attaching to the Ti atom. TiAl(2)O(2) (-) has two nearly degenerate Al-O-Ti-O-Al chain structures that can be considered as cis and trans forms. TiAl(2)O(3) (-) has two low-lying isomers, kite structure and book structure. The structures of these clusters indicate that the Ti atom tends to bind to more O atoms.


Slow, continuous enzyme replacement via spinal CSF in dogs with the paediatric-onset neurodegenerative disease, MPS IIIA.
Intra-cerebrospinal fluid (CSF) injection of recombinant human lysosomal enzyme is a potential treatment strategy for several neurodegenerative lysosomal storage disorders including Sanfilippo syndrome (Mucopolysaccharidosis type IIIA; MPS IIIA). Here we have utilised the MPS IIIA Huntaway dog model to compare the effectiveness of the repeated intermittent bolus injection strategy being used in the trials with an alternate approach; slow, continual infusion of replacement enzyme (recombinant human sulphamidase; rhSGSH) into the spinal CSF using a SynchroMed II® pump attached to a spinal infusion cannula. The ability of each enzyme delivery strategy to ameliorate lesions in MPS IIIA brain was determined in animals treated from ∼three- to six-months of age. Controls received buffer or no treatment. Significant reductions in heparan sulphate (primary substrate) were observed in brain samples from dogs treated via either cisternal or lumbar spinal CSF bolus injection methods and also in slow intra-spinal CSF infusion-treated dogs. The extent of the reduction differed regionally. Pump-delivered rhSGSH was less effective in reducing secondary substrate (GM3 ganglioside) in deeper aspects of cerebral cortex, and although near-amelioration of microglial activation was seen in superficial (but not deep) layers of cerebral cortex in both bolus enzyme-treated groups, pump-infusion of rhSGSH had little impact on microgliosis. While continual low-dose infusion of rhSGSH into MPS IIIA dog CSF reduces disease-based lesions in brain, it was not as efficacious as repeated cisternal or spinal CSF bolus infusion of rhSGSH over the time-frame of these experiments.


Effectiveness, User Engagement and Experience, and Safety of a Mobile App (Lumi Nova) Delivering Exposure-Based Cognitive Behavioral Therapy Strategies to Manage Anxiety in Children via Immersive Gaming Technology: Preliminary Evaluation Study.
Childhood anxiety disorders are a prevalent mental health problem that can be treated effectively with cognitive behavioral therapy, in which exposure is a key component; however, access to treatment is poor. Mobile-based apps on smartphones or tablets may facilitate the delivery of evidence-based therapy for child anxiety, thereby overcoming the access and engagement barriers of traditional treatment. Apps that deliver therapeutic content via immersive gaming technology could offer an effective, highly engaging, and flexible treatment proposition. In this paper, we aim to describe a preliminary multi-method evaluation of Lumi Nova, a mobile app intervention targeting mild to moderate anxiety problems in children aged 7-12 years using exposure therapy delivered via an immersive game. The primary objective is to evaluate the effectiveness, user engagement and experience, and safety of the beta version of Lumi Nova. Lumi Nova was co-designed with children, parents, teachers, clinicians, game industry experts, and academic partnerships. In total, 120 community-based children with mild to moderate anxiety and their guardians were enrolled to participate in an 8-week pilot study. The outcome measures captured the app's effectiveness (anxiety symptoms, child-identified goal-based outcomes, and functional impairment), user engagement (game play data and ease-of-use ratings), and safety (mood ratings and adverse events). The outcome measures before and after the intervention were available for 30 children (age: mean 9.8, SD 1.7 years; girls: 18/30, 60%; White: 24/30, 80%). Additional game play data were automatically generated for 67 children (age: mean 9.6, SD 1.53 years; girls: 35/67, 52%; White: 42/67, 63%). Postintervention open-response data from 53% (16/30) of guardians relating to the primary objectives were also examined. Playing Lumi Nova was effective in reducing anxiety symptom severity over the 8-week period of game play (t29=2.79; P=.009; Cohen d=0.35) and making progress toward treatment goals (z=2.43; P=.02), but there were no improvements in relation to functional impairment. Children found it easy to play the game and engaged safely with therapeutic content. However, the positive effects were small, and there were limitations to the game play data. This preliminary study provides initial evidence that an immersive mobile game app may safely benefit children experiencing mild to moderate anxiety. It also demonstrates the value of the rigorous evaluation of digital interventions during the development process to rapidly improve readiness for full market launch.


Surface-layer protein from Caulobacter crescentus: expression, purification and X-ray crystallographic analysis.
Protein surface layers are self-assembling, paracrystalline lattices on the surface of many prokaryotes. Surface-layer proteins have not benefited from widespread structural analysis owing to their resistance to crystallization. Here, the successful expression of a truncated version of RsaA, the surface-layer protein from Caulobacter crescentus, from a Caulobacter protein-expression system is reported. The purification, crystallization and initial X-ray diffraction analysis of the truncated RsaA, the largest surface-layer protein studied to date and the first from a Gram-negative bacterium, are also reported.


Efficacy of Ascorbic Acid (Vitamin C) and/N-Acetylcysteine (NAC) Supplementation on Nutritional and Antioxidant Status of Male Chronic Obstructive Pulmonary Disease (COPD) Patients.
Antioxidant therapy has a potential to be introduced as therapeutic modality for chronic obstructive pulmonary disease (COPD) patients. This study aimed to determine the effect of antioxidant supplementation [ascorbic acid and N-Acetylcysteine (NAC)] on nutritional and antioxidant status in male COPD patients. A parallel and single blind randomised controlled clinical trial (RCT) was conducted at two medical centers in Kuala Lumpur, Malaysia. Seventy-nine subjects were recruited and randomly divided into four trial arms (i.e., NAC, vitamin C, NAC+vitamin C and control groups) for six mo. The primary outcome was changes in body mass index by estimating power of 90% and significance level of p<0.05. Repeated Measure ANOVA showed that there was a significant interaction effect on BMI (p=0.046) and carbohydrate intake (p=0.030), especially in the NAC group. Plasma glutathione (GSH) increased significantly in all intervention groups, especially in vitamin C (p=0.005). A single supplementation of NAC or vitamin C improved nutritional and antioxidant status of subjects.


Resonance Rayleigh scattering for detection of proteins in HPLC.
An HPLC-resonance Rayleigh scattering (RRS) (HPLC-RRS) detection system is described for separation and detection of proteins. This system is based on the modification of a commercial HPLC instrument involving the addition of a pump and a T-shaped interface, and a common fluorescence detector was used for detection. The detection principle is based on the change of RRS intensity of the ion-association complex formed from biebrich scarlet (BS) and protein. The RRS signal was detected at lambdaex=lambdaem=376 nm. The utility of the presented method was demonstrated by the separation and determination of four proteins involving cytochrome (Cyt-c), lysozyme (Lys), HSA, and gamma-globulin (gamma-Glo). An LOD of 0.2-1.0 microg/mL was reached and a linear range was found between peak area and concentration in the range of 0.20-3.0 microg/mL for Cyt-c, 0.25-2.5 microg/mL for Lys, 1.5-10 microg/mL for HSA, and 2.0-15 microg/mL for gamma-Glo, with linear regression coefficients all above 0.99. The method presented has been applied to determine HSA and gamma-Glo in human serum samples synchronously.


A generalized model of gene dosage and dominant negative effects in macromolecular complexes.
Most genes and their corresponding products are supposed to be involved in genetic or biochemical interactions. A typical example is provided by macromolecular complexes, which may contain tens of proteins in defined stoichiometric proportions. Stoichiometric imbalances in such complexes can be a source of abnormal phenotypes. Comparable effects can also arise from negative dominance/transdominance, even though the underlying mechanisms are different. Here I propose a general yet simple biochemical model accounting for the effects of dosage changes and weak dominant/transdominant negative mutations in macromolecular complexes. The molecular alterations studied are predicted to lead to synergistic effects that can drive total multimer concentration and/or activity in a multiple heterozygote below a critical threshold required to ensure a normal phenotype, thus providing an explanation for the phenomenon of unlinked noncomplementation or nonallelic noncomplementation. The model also helps in understanding the basis of heterosis and the long-term consequences of gene dosage alterations and weak dominant/transdominant negative effects. Indeed, it can explain the observed extensive retention of paralogs in polyploids. Finally, because the effects of weak single-gene alterations can escape selection, they may accumulate in the population. This situation has important evolutionary consequences and may eventually lead to reproductive isolation and speciation.-Veitia, R. A. A generalized model of gene dosage and dominant negative effects in macromolecular complexes.


Chemical modification of SWNT alters in vitro cell-SWNT interactions.
Single-walled carbon nanotubes (SWNT) have been the focus of considerable attention as a material with extraordinary mechanical and electrical properties. SWNT have been proposed in a number of biomedical applications, including neural, bone, and dental tissue engineering. In these applications, it is clear that surrounding tissues will come into surface contact with SWNT composites, and compatibility between SWNT and host cells must be addressed. This investigation describes the gross physical and chemical effects of different SWNT preparations on in vitro cell viability and metabolic activity. Three different SWNT preparations were analyzed: as purchased (AP-NT), purified (PUR-NT), and functionalized with glucosamine (GA-NT), over concentrations of 0.001-1.0% (wt/vol). With the exception of the lowest SWNT concentrations, increasing concentrations of SWNT resulted in a decrease of cell viability, which was dependent on SWNT preparation. The metabolic activity of 3T3 cells was also dependent on SWNT preparation and concentration. These investigations have shown that these SWNT preparations have significant effects on in vitro cellular function that cannot be attributed to one factor alone, but are more likely the result of several unfavorable interactions. Effects, such as destabilizing the cell membrane, soluble toxic contaminants, and limitations in mass transfer as the SWNT coalesce into sheets, may all play a role in these interactions. Using comprehensive purification processes and modifying the NT-surface chemistry to introduce functional groups or reduce hydrophobicity or both, these interactions can be significantly improved.


Health Insurance Trends in United States Living Kidney Donors (2004 to 2015).
Some transplant programs consider the lack of health insurance as a contraindication to living kidney donation. Still, prior studies have shown that many adults are uninsured at time of donation. We extend the study of donor health insurance status over a longer time period and examine associations between insurance status and relevant sociodemographic and health characteristics. We queried the United Network for Organ Sharing/Organ Procurement and Transplantation Network registry for all living kidney donors (LKDs) between July 2004 and July 2015. Of the 53 724 LKDs with known health insurance status, 8306 (16%) were uninsured at the time of donation. Younger (18 to 34 years old), male, minority, unemployed, less educated, unmarried LKDs and those who were smokers and normotensive were more likely to not have health insurance at the time of donation. Compared to those with no health risk factors (i.e. obesity, smoking, hypertension, estimated glomerular filtration rate <60, proteinuria) (14%), LKDs with 1 (18%) or ≥2 (21%) health risk factors at the time of donation were more likely to be uninsured (p < 0.0001). Among those with ≥2 health risk factors, blacks (28%) and Hispanics (27%) had higher likelihood of being uninsured compared to whites (19%; p < 0.001). Study findings underscore the importance of providing health insurance benefits to all previous and future LKDs.


Correlation energy functional and potential from time-dependent exact-exchange theory.
In this work we studied a new functional for the correlation energy obtained from the exact-exchange (EXX) approximation within time-dependent density functional theory. Correlation energies have been calculated for a number of different atoms showing excellent agreement with results from more sophisticated methods. These results lose little accuracy by approximating the EXX kernel by its static value, a procedure which enormously simplifies the calculations. The correlation potential, obtained by taking the functional derivative with respect to the density, turns out to be remarkably accurate for all atoms studied. This potential has been used to calculate ionization potentials, static polarizabilities, and van der Waals coefficients with results in close agreement with experiment.


Long-term persistence of nonpathogenic clonal chromosome abnormalities in donor hematopoietic cells after allogeneic stem cell transplantation.
We describe the cases of two unrelated patients who exhibited multiple chromosomal abnormalities in donor cells after allogeneic peripheral blood stem cell transplantation (PBSCT). The patients were diagnosed with chronic myeloid leukemia and chronic lymphocytic leukemia, respectively, and both underwent nonmyeloablative conditioning with cyclophosphamide and fludarabine followed by PBSCT from their HLA-matched opposite-sex siblings. Post-transplant bone marrow cytogenetics showed full engraftment, and the early post-transplant studies demonstrated only normal donor metaphases. Subsequent studies of both patients, however, revealed a population of metaphase cells with abnormal, but apparently balanced, donor karyotypes. Chromosome studies performed on peripheral blood cells collected from both donors after transplantation were normal. Both patients remained in clinical remission during follow-up of approximately 8 years in one case, and 6 years in the other case, despite the persistence of the abnormal clones. Chromosomal abnormalities in residual recipient cells after bone marrow or PBSCT are not unusual. In contrast, only rare reports of chromosome abnormalities in donor cells exist, all of which have been associated with post-bone marrow transplant myelodysplastic syndrome or acute leukemias. The present cases demonstrate the rare phenomenon of persistent clonal nonpathogenic chromosome aberrations in cells of donor origin.


[Experimental study of ta chengchi tang decoction for relieving lung injury during acute necrotizing pancreatitis].
The objectives of the study were to investigate the changes of leukocyte adhesiveness and tumor necrosis factor (TNF) in the early stage of acute necrotizing pancreatitis (ANP), to go further into the relation of those changes to lung injury of ANP, and to evaluate the prohibitive effect of ta chengchi tang Decoction on leukocyte adhesion and TNF secretion. 14 canines were randomly divided into 3 groups. The treatment group (n = 5): experimental ANP model treated with ta chengchi tang Decoction; the saline control group (n = 5): experimental ANP model treated with normal saline; and the sham operative group (n = 4). The adhesion of leukocytes to vascular endothelial cells and TNF activities in serum and in bronchi-alveolar lavage fluid (BALF) were observed and measured, and the degrees of lung injury were assessed. The results showed that the adhesiveness of leukocytes was markedly increased and the activities of TNF were elevated in peripheral blood and BALF during ANP; that the leukocyte adhesiveness in the treatment group was weaker than that in the saline group (P < 0.05) and the serum and BALF TNF activities were also significantly lower than those in the saline group (P < 0.01); and that the degrees of lung injury in the treatment group were significantly milder than those in the saline group. These indicate that the increased leukocyte adhesiveness and hypersecretion of TNF take part in the pathogenesis of lung injury in ANP, and ta chengchi tang Decoction is demonstrated an efficacious medicine for alleviating the degree of lung injury mediated by both the leukocyte adhesion and TNF in ANP.


Incidence of cryptosporidiosis in immunodeficient cancer patients in Egypt.
A coproprotozoal study was carried out on 63 patients suffering from malignancy. The majority had cancer of haemopoietic system. All patients were under chemotherapy and included: Group A (33 children) and Group B (30 adults) of whom 20 immunocompetent diarrhoeic patients of matched age and sex were considered as controls. Stool samples were examined by merthiolate iodine-formaldehyde concentration technique (MIF). Modified Zeihl-Neelsen (ZN) stain was performed for Cryptosporidium oocysts. Detection of Cryptosporidium coproantigen by enzyme-linked immunoassay test (Ridascreen test), was used. Immunoglobulins (IgG, IgM, IgE & IgA), C3, C4 and CD4:CD8 ratio, were measured. According to their levels 25 out of 63 patients had both humoral and cellular immunodeficiency. The incidence of Cryptosporidium in cancer patients was 23.8%, while it was 37.7% and 91% in children and adults immunodeficient patients, respectively. ZN stain was able in diagnosed Cryptosporidium in 13 out of 35 immunodeficient cases while ELISA detected only 11 cases. Cryptosporidium infection in immunodeficient cancer patients had significantly more frequent and prolonged duration of diarrhoea than in negative ones.


Effects of presacral neurectomy on pelvic pain in women with and without endometriosis.
Presacral neurectomy (PSN) is used in treatment of central chronic pelvic pain (CPP); however, the confounding effect of concomitant resection of endometriosis remains uncertain. This study was undertaken to evaluate and compare the effectiveness of presacral neurectomy (PSN) in the presence and absence of endometriosis. Twenty-three women with midline CPP (age 30.3 +/- 7.9, range 21-46) unresponsive to medical therapy were recruited to the study. Endometriosis was absent in seven and present in sixteen subjects. Laparoscopic PSN using a harmonic scalpel was performed in all subjects; simultaneous excision of endometriotic lesions was also carried out in subjects with endometriosis. Intensity of dysmenorrhoea and pelvic pain was measured by visual analogue pain scale (VAPS) at 3 and 12 months postoperatively. Dysmenorrhoea decreased at 3 months by 75% (P = 0.018) in those without endometriosis and by 78% (P = 0.001) in those with endometriosis. At 12-months, dysmenorrhea increased in women with endometriosis (P = 0.008), but not in those without endometriosis. Pelvic pain not related to menses decreased by 67% (P = -0.0007) and by 87% (P = 0.028), respectively, in women with and without endometriosis. Dyspareunia, declined dramatically at 3 and 12 months to a median score of 0 (the majority of subjects had no discomfort; P < 0.001); the change in dyspareunia between 3 and 12 months was in favour of patients without endometriosis (P = 0.02) PSN using a harmonic scalpel results in long-term pain relief, especially in patients without endometriosis.


Rice embryo globulins: amino-terminal amino acid sequences, cDNA cloning and expression.
A globulin fraction prepared from rice embryos contained polypeptides or polypeptide groups of 49 kDa (designated REG1), 46 kDa (designated REG2), about 35 kDa, 32 kDa and 25 kDa. The amino-terminal sequences of REG1 and the major polypeptide in the 35-kDa group were identical, suggesting that the REG1 polypeptide undergoes partial proteolytic processing that removes a carboxy-terminal region. A cDNA clone, designated pcREG2, encoding REG2 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence of REG2 was found to be 68% identical to that of the maize GLB2 globulin. Reg2 mRNA was present at high levels during embryo development for up to 14 days after flowering (DAF). Lower levels were found 20 DAF when the maturation of embryos was almost completed, and at the dry mature stage. Reg2 mRNA almost disappeared upon imbibition of isolated dry mature embryos but it was re-induced at a low level by further treatment with ABA. The expression of Reg2 was not induced by ABA in suspension-cultured cells, unlike that of Osem, one of the late embryogenesis abundant protein (LEA) genes.


"Kiss myAsthma": Using a participatory design approach to develop a self-management app with young people with asthma.
Objective: Young people with asthma often lack engagement in self-management. Smartphone apps offer an attractive, immediate method for obtaining asthma information and self-management support. In this research we developed an evidence-based asthma app tailored to young peoples needs, created using a participatory design approach to optimize user engagement. This paper describes the participatory design process. Methods: This multi-phased research included concept generation and ideation of app design by young people with asthma, and development of asthma information by the research team. Clinical review was sought regarding safety and accuracy of app content. Participants suggestions for improvement and any problems with the app were logged throughout. Our young co-designers were invited back to test a high fidelity prototype app using a "think aloud" process and completed a usability questionnaire. Results: Twenty asthma patients aged 15-24 years contributed to the initial app design. Three respiratory specialists and two pharmacists suggested minor corrections to clinical terminology in the app which were all incorporated. Nine co-designers acted as expert reviewers of the prototype app, of whom eight completed a usability questionnaire. Median usability scores (maximum score 6) indicated high satisfaction with app content, usefulness and ease of use [median item score 5.3 (range 4.7-6.0)]. All feedback was incorporated to create an updated prototype app. Conclusions: A clinically sound asthma app has been developed which is considered highly acceptable to the young co-designers. A six-week test of the engagement, acceptability, and usefulness of the app in young people not involved in the participatory design will follow.


The relationship between apolipoprotein E polymorphism, lipoprotein (a) and fatty liver disease.
The apolipoprotein E (apo E) gene is known to affect plasma lipoprotein-lipid levels, and lipoprotein (a) [Lp(a)] is an atherogenic lipoprotein that is negatively correlated with triglyceride level. The purpose of this study was to investigate the relationship between the apo E genotype, plasma Lp(a) level, and fatty liver. A cross-sectional study was performed on 711 subjects who were diagnosed as having fatty liver by ultrasonography and on 711 sex- and age-matched control subjects. Apo E genotype, plasma Lp(a) concentrations, and serum levels of total cholesterol, triglyceride, LDL-cholesterol, and HDL-cholesterol were measured. Fatty liver subjects were found to have significantly higher triglyceride, and lower HDL-cholesterol levels. According to the multivariate logistic regression analysis, the odds ratios for fatty liver were 2.254 (95% CI 1.697-2.993) for hypertriglyceridemia, 1.841 (95% CI 1.449-2.339) for a high LDL-cholesterol level, 1.509 (95% CI 1.112-2.046) for a low HDL-cholesterol level, and 0.364 (95% CI 0.194-0.684) for a high HDL-cholesterol level. The odds ratios of epsilon4 and a high plasma Lp(a) level for fatty liver were 0.631 (95% CI 0.468-0.850) and 0.607 (95% CI 0.470-0.784) respectively. Our results suggest that the epsilon4 allele of apo E and plasma Lp(a) concentration may be associated with the pathogenesis of fatty liver.


Atmospheric Pressure Vaporization Mechanism for Coupling a Liquid Phase with Electron Ionization Mass Spectrometry.
A novel liquid chromatography-mass spectrometry (LC-MS) interfacing concept is presented and discussed. The new interface, called liquid-EI (LEI), is based on electron ionization (EI) but, differently from any previous attempt, the vaporization of solutes and mobile phase takes place at atmospheric pressure into a specifically designed region, called "vaporization microchannel", before entering the high-vacuum ion source. The interface is completely independent from the rest of the instrumentation and can be adapted to any gas chromatography/mass spectrometry (GC/MS) system, as an add-on for a rapid LC-MS conversion. Pressure drop and temperature gradient between LC and MS were considered to enhance the analyte response and reduce band broadening and/or solute carryovers. A fused silica liner, placed inside the vaporization microchannel, acts as an inert vaporization surface speeding up the gas-phase conversion of large molecules while lessening possible memory effects. The liner is easily replaceable for a quick and extremely simple interface maintenance. Proof of concept and detailed description of the interface are here presented.


Effect of calcium on the surface properties of phospholipid monolayers with respect to surfactant formulations in respiratory distress syndrome.
The effects of calcium, in the form of calcium chloride, at concentrations of 5 and 20 mM, were studied on the surface properties of physiologic relevance to specialised biomaterials which replace lung surfactant in Respiratory Distress Syndrome. The dynamic surface pressure, re-spreading ratio, compressibility, hysteresis area and recruitment index of pure films of the main phospholipids of pulmonary surfactant namely dipalmitoyl phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine and of binary mixed films of these phospholipids in the ratio of 2:3 were studied both in the presence and absence of calcium by in vitro analysis using a Wilhelmy balance. Surface excess films, of each of the surfactant systems, with initial concentration 15 A2 molecule were compressed at the rate of 50 seconds/cycle past collapse till a compression ratio of 4:1. The presence of 5 mM calcium caused a significant decrease in compressibility (p < 0.05 Mann-Whitney U test) of all the surfactant monolayer films. No further benefit was obtained by adding 20 mM calcium over that of 5 mM calcium. A significant beneficial effect of calcium (p < 0.05 Mann-Whitney U test) on film stability was observed when analysing the materials in a pulsating bubble surfactometer, in which liposomal suspensions of 1% concentration in the presence of 5 mM calcium were pulsated at a high frequency of 40 cycles per minute, corresponding to the respiratory frequency of neonates. The ultrastructure of the liposomal suspensions were also studied using cryogenic scanning electron microscopy and longitudinal micro-tubular structures were found on addition of 5 mM calcium, which could have resulted in the improved performance of the exogenous surfactants with respect to compressibility and stability.


Dopamine agonist treatment of schizophrenia with N-propylnorapomorphine.
We administered N-propylnorapomorphine, a potent aporphine-family dopamine (DA) agonist, to schizophrenic patients with active psychotic symptoms. After acute administration a significant antipsychotic action of N-propylnorapomorphine, maximal at an oral dose of 19 mg, was noted. The antipsychotic action predominated in subjects with neuroleptic-responsive symptoms, not in neuroleptic-nonresponders. However, when N-propylnorapomorphine was administered on a subchronic dosage schedule, no antipsychotic effect occurred. These observations suggest a rapid-onset tolerance phenomenon of psychosis to N-propylnorapomorphine and are consistent with results from preclinical experiments. These data support the idea that the acute antipsychotic response of DA agonists is mediated by the DA autoreceptor but fail to provide evidence for the potential clinical usefulness of this treatment approach.


Individualized Constellation of Killer Cell Immunoglobulin-Like Receptors and Cognate HLA Class I Ligands that Controls Natural Killer Cell Antiviral Immunity Predisposes COVID-19.
Background: The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection causes coronavirus disease-2019 (COVID-19) in some individuals, while the majority remain asymptomatic. Natural killer (NK) cells play an essential role in antiviral defense. NK cell maturation and function are regulated mainly by highly polymorphic killer cell immunoglobulin-like receptors (KIR) and cognate HLA class I ligands. Herein, we tested our hypothesis that the individualized KIR and HLA class I ligand combinations that control NK cell function determine the outcome of SARS-CoV-2 infection. Methods: We characterized KIR and HLA genes in 200 patients hospitalized for COVID-19 and 195 healthy general population controls. Results: The KIR3DL1+HLA-Bw4+ [Odds ratio (OR) = 0.65, p = 0.03] and KIR3DL2+HLA-A3/11+ (OR = 0.6, p = 0.02) combinations were encountered at significantly lower frequency in COVID-19 patients than in the controls. Notably, 40% of the patients lacked both of these KIR+HLA+ combinations compared to 24.6% of the controls (OR = 2.04, p = 0.001). Additionally, activating receptors KIR2DS1+KIR2DS5+ are more frequent in patients with severe COVID-19 than patients with mild disease (OR = 1.8, p = 0.05). Individuals carrying KIR2DS1+KIR2DS5+ genes but missing either KIR3DL1+HLA-Bw4+ combination (OR = 1.73, p = 0.04) or KIR3DL2+HLA-A3/11+ combination (OR = 1.75, p = 0.02) or both KIR3DL1+HLA-Bw4+ and KIR2DL2+HLA-A3/11+ combinations (OR = 1.63, p = 0.03) were more frequent in the COVID-19 cohort compared to controls. Conclusions: The absence of KIR3DL1+HLA-Bw4+ and KIR3DL2+HLA-A3/11+ combinations presumably yields inadequate NK cell maturation and reduces anti-SARS-CoV-2 defense, causing COVID-19. An increased frequency of KIR2DS1+KIR2DS5+ in severe COVID-19 patients suggests vigorous NK cell response triggered via these activating receptors and subsequent production of exuberant inflammatory cytokines responsible for severe COVID-19. Our results demonstrate that specific KIR-HLA combinations that control NK cell maturation and function are underlying immunogenetic variables that determine the dual role of NK cells in mediating beneficial antiviral and detrimental pathologic action. These findings offer a framework for developing potential host genetic biomarkers to distinguish individuals prone to COVID-19.


ONO-1714, a new inducible nitric oxide synthase inhibitor, attenuates diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats.
Acute experimental pancreatitis (induced by cerulein) recently has been reported to cause marked diaphragmatic dysfunction, which may contribute to respiratory distress in this setting. In cerulein-induced acute pancreatitis, expression of inducible nitric oxide synthase is induced to produce a large amount of nitric oxide. Nitric oxide excessively produced has been implicated in diaphragmatic dysfunction induced by a variety of etiologies. The aims of the current study were, first, to examine whether nitric oxide overproduced through inducible nitric oxide synthase is involved in cerulein-induced impairment of diaphragmatic function, and second, if demonstrated, to assess effects of ONO-1714, an inducible nitric oxide synthase inhibitor, on diaphragmatic dysfunction associated with cerulein-induced acute pancreatitis. Prospective, randomized animal study. University research laboratory. Ninety-one male Sprague-Dawley rats, weighing 200-250 g. Rats were randomly divided into seven groups (n = 8 each): CONT-SAL, CAER-SAL, CONT-ONO, CAER-DEX, CAER-AMI, CAER-ONOhigh, and CAER-ONOlow. Groups labeled CAER received two consecutive intraperitoneal doses (50 microg/kg) of cerulein, whereas groups labeled CONT received two consecutive intraperitoneal injections of saline. Groups labeled SAL received intraperitoneal saline before cerulein or saline. The group labeled DEX received 2 mg/kg intraperitoneal dexamethasone, and the group labeled AMI received 100 mg/kg intraperitoneal aminoguanidine. The groups labeled ONO, ONOhigh, and ONOlow received ONO-1714 at 0.1 mg/kg, 0.1 mg/kg, and 0.03 mg/kg, respectively, before cerulein or saline. Diaphragmatic contractility and fatigability were assessed in vitro by using muscle strips excised from the costal diaphragms 6 hrs after the first dose of cerulein or saline. Expression of inducible nitric oxide synthase protein in the diaphragm was assessed by immunohistochemistry by using anti-inducible nitric oxide synthase antibody. Plasma concentrations of nitrite plus nitrate and diaphragmatic concentrations of malondialdehyde were measured. With another set of rats (n = 5 each group), diaphragmatic inducible nitric oxide synthase activity was determined. Twitch and tetanic tensions and tensions generated during fatigue trial were lower in group CAER-SAL than in group CONT-SAL. Cerulein increased diaphragmatic malondialdehyde and plasma nitrite plus nitrate concentrations. Positive immunostaining for inducible nitric oxide synthase protein was found in group CAER-SAL. Dexamethasone and aminoguanidine attenuated the diaphragmatic mechanical damages. A high dose of ONO-1714 attenuated cerulein-induced impairment of diaphragmatic contractility and endurance capacity, although a low dose of the drug failed to do so. Cerulein-induced diaphragmatic dysfunction was attributable, in part, to nitric oxide overproduced via inducible nitric oxide synthase. Pretreatment with ONO-1714 at a dose of 0.1 mg/kg attenuated diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats assessed by contractile profiles and endurance capacity. This beneficial effect of ONO-1714 may be attributable, in part, to inhibition of diaphragmatic lipid peroxidation induced by nitric oxide-derived free radicals.


[Enteral feeding in oncology].
Nutrition therapy is accepted to represent an important part of therapeutic strategy in cancer patients. In correlation to general clinical state, nutritional condition and anti-cancer therapy, nutrition has to be planned according to a stepwise concept. Under certain circumstances, enteral tube feeding is indicated. This procedure is affected by tumor lesions, digestion, resorption and tumor related metabolic disorders, and side effects of particulate anti-cancer therapy. In tumor patients, elemental (chemically defined) diets are predominant. Polymeric diets, however, occasionally represent another possible nutritional tool.


Globalization and the trends of medical technology trade in Turkey.
Medical technology trade is one of the most affected health areas by global regulations in the developing countries. The aim of the study is to examine recent changes in medical technology import and export and their results in Turkey. Data show that the total medical technology imports (MTI) increased from $ 34.6 million to $ 3427.9 million between 1970 and 2003. While MTI constituted 3.6% of total imports in 1970 and 1.3% in 1980, this ratio raised up to 4.9% in 2003. The ratio of MTI in total health expenditures were also increased from 7.6% in 1970 to 31.5% in 2003. Medical technology exports (MTE) have been increased from $ 0.9 million in 1970 to $ 303.2 million in 2003. The ratio of MTE to MTI increased from 2.7% to 13.9% between 1970 and 1990 and decreased after 1990, to 8.8%. Our study implied that the medical technology trade in Turkey has been negatively affected and in some respects differs from some other important industries in the globalization era. Nevertheless, detailed comparative studies in different developing countries such as China, Brazil, Mexico and India, are needed to explore the real state of medical technology trade, use and the effects of globalization on these topics.


Slow transit constipation: a disorder of pelvic autonomic nerves?
Slow transit constipation (STC) is a severe motility disorder, which in the majority of cases is of unknown etiology. In some, symptoms arise de novo in childhood, but a proportion of patients present in later life, including after pelvic surgery or childbirth. Our aims were: (1) to describe our current knowledge of the anatomy and function of the pelvic autonomic nerves with respect to colonic motility (experimental and observational studies); (2) to discuss evidence for pelvic nerve injury in STC arising after pelvic surgery or childbirth; and (3), on the basis that such patients are clinically indistinguishable from patients with chronic idiopathic STC, to evaluate whether there is evidence that pelvic autonomic neuropathy has an etiologic role in patients with chronic idiopathic STC. The outcome was as follows: (1) The clear importance of the pelvic autonomic nerves in colonic motor function is documented. (2) While there is an association between pelvic surgery and childbirth, and the onset of STC, there is little direct anatomical evidence that pelvic denervation occurs in these patients. However the phenotype of these patients is similar to results of experimental and observational studies. (3) Clinical, physiological, and histological similarities exist between patients whose symptoms arose following pelvic intervention and those whose symptoms arise de novo (idiopathic). We further present evidence for possible pathogenetic mechanisms underlying pelvic autonomic neuropathy in chronic idiopathic STC.


Gestational thyrotoxicosis with acute Wernicke encephalopathy: a case report.
A 35-year-old hyperthyroid woman who developed nausea, vomiting, tachycardia, nystagmus and mental disturbance, was referred to our hospital with a suspected diagnosis of thyroid storm. However, the thyroid gland was only slightly palpable, bruits were not audible, and exophthalmos was not present. Serum levels of thyroid hormone were increased, but TSH receptor antibodies were negative. Echography and color flow doppler ultrasonography revealed a slightly enlarged thyroid gland and a slightly increased blood flow, both of which were much less milder than those expected for severe hyperthyroid Graves' disease. Under the diagnosis of hyperthyroidism due to gestational thyrotoxicosis associated with Wernicke encephalopathy, vitamin B1 was administered on the first day of admission. Her consciousness became nearly normal on the second day except for slight amnesia. Her right abducent nerve palsy rapidly improved, but horizontal and vertical nystagmus, diminished deep tendon reflexes and gait ataxia improved only gradually. MRI findings of the brain were compatible with acute Wernicke encephalopathy. We concluded that history taking and physical findings are important to make a differential diagnosis of gestational thyrotoxicosis with acute Wernicke encephalopathy from Graves' thyroid storm, and that Wernicke encephalopathy should be treated as soon as possible to improve the prognosis.


Experience with surgical resection of heterotopic bone in spinal cord injury patients.
Extensive heterotopic ossification may occur after a spinal cord injury. Surgical removal of the heterotopic bone is considered only in patients who develop limitations of function or related skin pressure areas. In a prospective study of 21 patients, a total of 43 surgical procedures were performed to resect heterotopic bone. Long-term follow-up study was available for 18 patients who had 38 surgical procedures. This was a descriptive study since preoperative, surgical, and postoperative care varied slightly over time and were dependent on newer advances in surgical techniques and therapeutic agents. Primary complications included excessive bleeding, infection, fractures, and recurrence of the heterotopic ossification. Procedures are outlined that may reduce the incidence of postoperative infection. Fractures are related to the degree of regional osteoporosis. Recurrence is still a common problem, and any or all therapeutic agents that may reduce the extent of postoperative recurrence should be considered before surgical excision is planned.


Fibronectin and integrin alpha 5 play essential roles in the development of the cardiac neural crest.
Cardiac neural crest (CNC) plays a requisite role during cardiovascular development and defects in the formation of CNC-derived structures underlie several common forms of human congenital birth defects. Migration of the CNC cells to their destinations as well as expansion and maintenance of these cells are important for the normal development of the cardiac outflow tract and aortic arch arteries; however, molecular mechanisms regulating these processes are not well-understood. Fibronectin (FN) protein is present along neural crest migration paths and neural crest cells migrate when plated on FN in vitro; therefore, we tested the role of FN during the development of the CNC in vivo. Our analysis of the fate of the neural crest shows that CNC cells reach their destinations in the branchial arches and the cardiac outflow tract in the absence of FN or its cellular receptor integrin α5β1. However, we found that FN and integrin α5 modulate CNC proliferation and survival, and are required for the presence of normal numbers of CNC cells at their destinations.


Inactivation of trypsin-like proteases by depsipeptides of p-guanidinobenzoic acid.
A number of esters of p-guanidinobenzoic acid have been synthesized which contain a glycolyl peptide as the departing group. In the case of several enzymes such as trypsin and plasma kallikrein, depsipeptides were obtained which were considerably more reactive than the ethyl ester in inactivation of the protease by acyl-enzyme formation; the depsipeptide processing -CH2CO-Phe-NH2 as a leaving group displayed the highest reactivity. They were less effective in the case of urokinase, plasmin, and urinary kallikrein. Boar acrosin was very susceptible to inactivation by both ethyl and peptidyl esters. Depsipeptides possessing a longer peptide chain and a secondary carbon as a leaving group showed lower activities. The results demonstrate the productive use of the departing group region of protease active centers to obtain selectivity.


[The GHRH test in Turner's syndrome].
GHRH test was performed in 11 girls suffering from Turner's syndrome ranging in age from 5.6-13.5 years. GH peak resulted lower than 10 ng/ml in three subjects, who had also shown reduced GH values after two conventional pharmacological stimuli (L-dopa- and insulin-induced hypoglycemia) and a value of mean GH concentration over 24 hours lower than 3 ng/ml. Both GH peak and area under the curve were not correlated with height, height velocity, bone age/chronological age ratio, GH peak after conventional pharmacological stimuli and mean GH value of spontaneous secretion. The comparison with the results of GHRH test in other kinds of short stature evidenced in girls with Turner's syndrome the presence of GH values (peak and area under the curve) higher than those in subjects with "classical" GH deficiency, lower than those in "short normal stature" and similar to those in subjects with "non classical" GH deficiency. In conclusion, our data suggest, even if within a certain variability of the responses, a possible involvement of GH deficiency to the pathogenesis of short stature in Turner's syndrome, suggesting the existence of a prevalent hypothalamic nature of GH deficiency.


Expression and Functions of the CB2 Receptor in Human Leukocytes.
The cannabinoid CB2 receptor was cloned from the promyeloid cell line HL-60 and is notably expressed in most, if not all leukocyte types. This relatively restricted localization, combined to the absence of psychotropic effects following its activation, make it an attractive drug target for inflammatory and autoimmune diseases. Therefore, there has been an increasing interest in the past decades to identify precisely which immune cells express the CB2 receptor and what are the consequences of such activation. Herein, we provide new data on the expression of both CB1 and CB2 receptors by human blood leukocytes and discuss the impact of CB2 receptor activation in human leukocytes. While the expression of the CB2 mRNA can be detected in eosinophils, neutrophils, monocytes, B and T lymphocytes, this receptor is most abundant in human eosinophils and B lymphocytes. We also review the evidence obtained from primary human leukocytes and immortalized cell lines regarding the regulation of their functions by the CB2 receptor, which underscore the urgent need to deepen our understanding of the CB2 receptor as an immunoregulator in humans.


Macrophage responses to a Zr-based bulk metallic glass.
Bulk metallic glasses (BMGs) are considered to be a competitive candidate of biomedical materials, owing to their appealing mechanical properties and high thermal processability. Based on the established biosafety of the Zr-based BMGs, macrophage responses to (Zr₅₅ Al₁₀ Ni₅ Cu₃₀)₉₉ Y₁ (atomic percent) BMG were investigated in the present study, in comparison with Ti-6Al-4V alloy. The adhesion of RAW 264.7 macrophages to both alloys was found to be mediated by protein adsorption. The Zr-based BMG is capable of supporting regular adhesion and proliferation of macrophages, indicating its good biocompatibility, which agrees with previous findings using other mammalian cells. A lower degree of morphological activation was revealed on Zr-based BMG substrates than on Ti-6Al-4V substrates, which is evidenced by smaller spreading areas and less ruffling on cell surfaces. Smaller amount of proinflammatory cytokine, tumor necrosis factor-alpha, was secreted by macrophages cultured on Zr-based BMGs, which further confirms the lower level of inflammation induced by BMG than by Ti alloys.


[Two cases of neuroendocrine carcinoma of the rectum].
We report two cases of neuroendocrine carcinoma of the rectum. CASE 1: A 50s woman was diagnosed as rectal cancer and underwent anterior resection of the rectum and lymphnode dissection. The histological diagnosis was neuroendocrine carcinoma with peritoneal dissemination. She was treated with chemotherapy as an outpatient. One year later from the operation, multiple liver metastases were revealed and she died eight months later. CASE 2: A 50s man underwent endoscopic mucosal resection (EMR) of the rectum as rectal tumor and histological diagnosis was an early well-moderate deferenciated carcinoma and its cut-tend was unclear. He received a careful follow-up. One year later, a follow-up colonoscopy revealed a submucosal tumor in the lower rectum. He was diagnosised with local reccurence of rectal cancer, and then underwent an abdominoperineal resection of the rectum and lymphnode dissection. The histological diagnosis was poorly differenciated neuroendocrine carcinoma with lymphnode metastasis. Two months later from the operation, a local reccurence was revealed and he was treated with octreotide and irradiation.


Optimizing dissolved air flotation design and saturation.
Dissolved air flotation (DAF) of iron hydroxide precipitates at working pressures lower than 3 atm, using modified flotation units to improve the collection of fragile coagula, was studied. Conventional DAF flotation was studied as a function of saturation pressure in the absence and presence of surfactants in the saturator. Without surfactants, the minimum saturation pressure required for DAF to occur was found to be 3 atm. But, by lowering the air/water surface tension in the saturator, DAF was possible at a saturation pressure of 2 atm. This behavior was found to occur in both batch and pilot DAF operation tests and almost complete recovery of the precipitates was attained. Results are explained in terms of the minimum "energy" which has to be transferred to the liquid phase to form bubbles by a cavity phenomenon. Further, studies were conducted changing equipment design and feed bubbles size distribution (mixing micro and "mid-sized" bubbles). Thus, bubbles entrance position in the collision-adhesion zone ("capture" zone) was compared to bubble entrance position in the water flow inlet below the floating bed. A "mushroom" type diffuser was used for the "capture zone" experiment and better performance was obtained. Results are explained in terms of different mass transfer phenomena in the collection zone and in the separation zone. Finally, results obtained with the use of a column flotation cell working as normal DAF and with a wide bubble size range are presented. Results indicate good performance and some gains in process kinetics with middle size bubbles.


Clinical significance of surfactant protein D as a serum marker for evaluating pulmonary fibrosis in patients with systemic sclerosis.
To determine the clinical significance of surfactant protein D (SP-D), a useful marker for evaluating various lung diseases, in patients with systemic sclerosis (SSc) and to clarify any clinical significance between SP-D and KL-6, which is known to be correlated with pulmonary fibrosis (PF) in SSc patients. We used a specific enzyme-linked immunosorbent assay to measure serum SP-D levels in 83 patients with SSc and 31 healthy control subjects. The serum levels of SP-D were significantly higher in patients with SSc than in healthy controls (mean +/- SD 81.9+/-59.2 versus 34.8+/-13.7 ng/ml). Serum SP-D levels in patients with diffuse cutaneous SSc were significantly higher than those in patients with limited cutaneous SSc (98.8+/-72.1 versus 66.8+/-40.0 ng/ml). Serum SP-D levels in patients with PF were significantly elevated compared with those in patients without PF (99.7+/-64.1 versus 65.3+/-49.4 ng/ml). Moreover, the incidences of decreased percentage diffusing capacity for carbon monoxide and decreased percentage vital capacity were also significantly greater in patients with elevated SP-D levels than in those with normal levels (67% versus 43% and 36% versus 17%, respectively). There was a significant positive correlation between serum levels of SP-D and KL-6. Serum SP-D and KL-6 levels showed almost the same sensitivities and specificities in the diagnosis of PF (68% versus 73% and 70% versus 74%, respectively). These two markers also predicted PF to almost the same degree (31% versus 33%, respectively). These results suggest that SP-D, as well as KL-6, may be a useful serum marker for evaluating PF in patients with SSc.


[Tracheal intubation guided by fibrobronchoscopy in patients with difficult airway. Predictive factors of the outcome].
Tracheal intubation (TI) guided by fibrobronchoscopy (FB) is one of the essential techniques in the approach to the difficult airway (DAW). Few works have been published on the possible causes of TI failure with this procedure. This study aims to discover which factors could predict TI failure with FB. An observational and retrospective study in which the last 122 consecutive TIs guided by FB (between January 2000 and April 2008) performed by our group were included. A multivariate analysis of the factors that could influence in the outcome was conducted: cause of the DAW, TI pathway, type of endotracheal tube, elective or urgent indication of the procedure, sedation level and experience of the bronchoscopist. Tracheal intubation in individuals who are going to undergo surgical interventions accounts for 92.6% of the total. The most frequent indications of TI by FB were: limitation of neck movement (60 cases), airway stenosis (24), increase of soft tissues (13), narrow oral aperture (9), airway compression (6), and vocal cord paralysis (6). In 10 (8.2%) cases, TI by FB was not possible. The variables that best predicted IT failure in the multivariate analysis were profound sedation/ general anesthesia (OR = 12.2; 95% CI, 1.8-84; p = 0.01) and limited experience of the bronchoscopist (OR = 25.3; 95% CI, 3.5-181.8; p = 0.001). TI guided by FB performed by bronchoscopist is successful in more than 90% of the cases with DAW. The skill and experience of the bronchoscopist is one of the primary determining factors of success of the procedure. Profound sedation may condition TI guided by FB failure.


The contribution of [Ca2+]i to the slowing of relaxation in fatigued single fibres from mouse skeletal muscle.
1. The contribution of Ca2+ vs. cross-bridges to the slowing of relaxation in fatigue was studied in intact, single fibres dissected from a mouse foot muscle. Fatigue was produced by repeated 350 ms tetani. The free myoplasmic Ca2+ concentration ([Ca2+]i) was measured with indo-1. 2. The rate of [Ca2+]i decline after a tetanus was reduced in fatigue but this reduction appeared smaller than the decrease in relaxation speed. 3. Under control conditions, steady-state [Ca2+]i-tension curves were constructed from measurements of tetani at various stimulus frequencies. Measurements from the final part of fatigue runs, where both tetanic [Ca2+]i and tension fell relatively fast, were used to construct similar curves in fatigue. A comparison of these curves revealed a reduction of both the maximum Ca(2+)-activated tension and the myofibrillar Ca2+ sensitivity in fatigue. 4. Calcium-derived tension records were generated by converting [Ca2+]i signals during tetani into tension by means of the steady-state [Ca2+]i-tension relation. These records would represent a situation where tension responds to changes of [Ca2+]i without delays due to, for instance, cross-bridge attachment or detachment. The relaxation speed of the calcium-derived tension was similar in control and fatigue; the lag between the calcium-derived and the real tension was about twice as long in fatigue. 5. Both the decline of [Ca2+]i and tension were markedly faster after a 10 s pause elicited during fatigue runs at the time of maximum slowing of relaxation. The relaxation of the calcium-derived tension was also faster after the pause. 6. The function of the sarcoplasmic reticulum (SR) Ca2+ pumps was analysed by plotting the rate of [Ca2+]i decline vs. [Ca2+]i during the tail of elevated [Ca2+]i after tetani. This analysis showed approximately a sevenfold reduction of the pump rate in fatigue. 7. In conclusion, the rate of [Ca2+]i decline after a tetanus is reduced in fatigue probably due to impaired SR Ca2+ pumping. This is not the cause of the slowed relaxation because it is counteracted by a reduced myofibrillar Ca2+ sensitivity. Thus, the slowing of relaxation in fatigued mouse muscle fibres would reflect slowed cross-bridge kinetics.


[Growth and photosynthetic characteristics of field-grown Coffea arabica under different watering and fertilization managements].
Straw mulching and drip irrigation have been widely used for coffee production. In order to know if these techniques are suitable in Yunnan, five-year-old coffee plants were separated into 8 groups to grow in the field under 2 fertilization rates (low and high) and 4 watering regimes: dry straw mulching (M), drip irrigation (I), drip irrigation + straw mulching (MI) and control treatment (CK), and the effects of various watering and fertilization treatments on the growth and photosynthetic characteristics of Coffea arabica with the density of 4500 individuals per hectare were measured. The results showed that during one year's growth period, the coffee plants had two growth peaks, one was in May when the rainy season just begun, and the another was in August to September, the middle of rainy season. Higher fertilization promoted the relative height and length growth rates of the branches, but watering treatments hadn't significant effects on them. In dry season, watering significantly promoted the Pn, gs, Tr and WUE, while water status had no significant influence on the internal fluorescence features of PSII. In wet season, high-fertilized plants had a higher leaf nitrogen content and Pn than low-fertilized plants, and Pn was positively correlated with leaf nitrogen content. In both seasons, higher fertilization increased the WUE regardless of the watering treatments. At the same time, the high-fertilization treatment significantly alleviated diurnal photoinhibition, companying with a higher energy utilization through photochemistry and a higher energy dissipation through xanthophyll cycle, which appears that high-fertilized coffee plants have the mechanism to acclimate to strong light environment. All the results indicated that wet season is the optimum time for the photosynthesis and growth of C. arabica, and C. arabica needs a high fertilization investment and fine watering managements during its whole growth period. Among the three man-made watering treatments, MI was the best, M and I had the nearly same effects but the former was more practical than the latter.


Ubiquitin-proteasome system involvement in Huntington's disease.
Huntington's disease (HD) is a genetic autosomal dominant neurodegenerative disease caused by the expansion of a CAG repeat in the huntingtin (htt) gene. This triplet expansion encodes a polyglutamine stretch (polyQ) in the N-terminus of the high molecular weight (348-kDa) and ubiquitously expressed protein htt. Normal individuals have between 6 and 35 CAG triplets, while expansions longer than 40 repeats lead to HD. The onset and severity of the disease depend on the length of the polyQ tract: the longer the polyglutamine stretch (polyQ) is, the earlier the disease begins and the more severe the symptoms are. One of the main histopathological hallmarks of HD is the presence of intraneuronal proteinaceous inclusion bodies, whose prominent and invariant feature is the presence of ubiquitin (Ub); therefore, they can be detected with anti-ubiquitin and anti-proteasome antibodies. This, together with the observation that mutations in components of the ubiquitin-proteasome system (UPS) give rise to some neurodegenerative diseases, suggests that UPS impairment may be causative of HD. Even though the link between disrupted Ub homeostasis and protein aggregation to HD is undisputed, the functional significance of these correlations and their mechanistic implications remains unresolved. Moreover, there is no consistent evidence documenting an accompanying decrease in levels of free Ub or disruption of Ub pool dynamics in neurodegenerative disease or models thus suggesting that the Ub-conjugate accumulation may be benign and just underlie lesion in 26S function. In this chapter we will elaborate on the different studies that have been performed using different experimental approaches, in order to shed light to this matter.


Spin-label ESR with nanochannels to improve the study of backbone dynamics and structural conformations of polypeptides.
Nanochannels of mesoporous silica materials were previously found useful for reducing the tumbling motion of encapsulated biomolecules while leaving the biomolecular structure undisturbed. Here we show that experiments of cw-ESR distance measurement in nano-confinement can benefit immediately from the above mentioned features of sufficiently slow molecular tumbling, enabling more accurate determination of interspin distances throughout the temperature range, from 200 to 300 K. A 26-residue prion protein peptide, which can fold into either a helical or hairpin structure, as well as its variants, are studied by using ESR. By comparing the spectra obtained in vitrified bulk solutions vs. mesopores, the spectra from the latter display typical slow-motional lineshapes, thereby enabling dipolar anisotropy to be unambiguously revealed throughout the temperature range, whereas the spectra from the former are dominated by the disordering of the side chain and the rotational tumbling of the peptide. The spectral changes regarding the two secondary structures in nano-confinement are found to show a strong correlation with the dynamic properties of the backbones. The effect of viscosity agent perturbation on the motion of an R1 nitroxide side chain, a commonly employed probe, could be substantial in a bulk solution condition, though it is absolutely absent in nanochannels. Under nano-confinement, the probe is proven sufficiently sensitive to the backbone motions. Overall, the distance distributions determined from the mesopore studies not only describe the conformational structures (by average distances), but also the backbone dynamics (by distribution widths) of the spin-labeled peptides.


Evaluation of nicotinamide microemulsion on the skin penetration enhancement.
This study purposed to evaluate a microemulsion containing nicotinamide for its characteristics, stability, and skin penetration and retention comparing with a solution of nicotinamide in 2:1 mixture of water and isopropyl alcohol (IPA). The microemulsion system was composed of 1:1 mixture of Span80 and Tween80 as a surfactant mixture, isopropyl palmitate (IPP) as an oil phase, and 2:1 mixture of water and IPA as an aqueous phase. Nicotinamide microemulsion was prepared by dissolving the active in the aqueous phase before simply mixing with the other components. It was determined for its characteristics and stability under various conditions. The skin penetration and retention studies of nicotinamide microemulsion and solution were performed by modified Franz diffusion cells, using newborn pig skin as the membrane. The results showed that nicotinamide microemulsion could be obtained as clear yellowish liquid, was water-in-oil (w/o) type, possessed Newtonian flow, and exhibited physicochemical stability when kept at 4 °C and room temperature (≈30 ± 2 °C) during 3 months. From the skin penetration data, the microemulsion could enhance the skin penetration of nicotinamide comparing with the solution. Additionally, nicotinamide microemulsion could provide much higher amount of skin retention than that of skin penetration, resulting in suitability for a cosmeceutical product.


The primary structure of sturgeon prolactin: phylogenetic implication.
The complete amino acid sequence of prolactin (PRL) from a chondrostean species, the sturgeon (Acipenser gueldenstaedti), has been determined. Sturgeon PRL was isolated from the pituitary glands by gel filtration on a Sephadex G-25 column and high-performance liquid chromatography on a reverse-phase column following acid-acetone extraction. Sturgeon PRL was identified by immunoblot reactivity using antisera against salmon and ovine PRL. It consists of 204 amino acid residues, which is the largest among known PRLs, and contains three disulfide bonds corresponding to those of tetrapod PRLs. Sequence comparison with PRLs from other vertebrates revealed that sturgeon PRL has slightly higher sequence identities (35-46%) with teleost PRLs than with tetrapod PRLs (30-40%). These structural characteristics imply that an ancestor of the ray-finned fishes had PRL with three disulfide bonds and at some point after divergence of Chondrostei, the disulfide bond in the amino-terminus of PRL was lost.


Curcumin Targeting NF-κB/Ubiquitin-Proteasome-System Axis Ameliorates Muscle Atrophy in Triple-Negative Breast Cancer Cachexia Mice.
Curcumin is a polyphenol plant-derived compound with anti-inflammatory, antioxidant stress, and anticancer properties that make it have the potential to treat cancer cachexia. However, the role of it in breast cancer cachexia remains unclear. The 4T1 cells were subcutaneously injected into BALB/c mice to induce breast cancer cachexia. After tumor formation, the animals were divided into groups and given curcumin or saline interventions. The therapeutic effect of curcumin on breast cancer cachexia was characterized by tumor growth, changes in body mass and gastrocnemius mass, muscle function test, histopathology, and serum nutrition indexes. Mitochondrial function in muscle tissue was observed by transmission electron microscopy and ATP detection, muscle inflammatory factors were detected by ELISA, muscle differential metabolites were detected by 1HNMR metabolomics, and the muscle tissue ubiquitination levels and NF-KB expression were also analyzed by RT-qPCR and Western blot. Dynamic in vivo bioluminescence imaging find that curcumin inhibited the growth of tumor in triple-negative breast cancer- (TNBC-) bearing mice, slowed down the loss of body weight and gastrocnemius weight, corrected the mitochondrial dysfunction and malnutrition status, and also significantly improved skeletal muscle function. ELISA analysis found that the level of inflammatory factors in muscle tissue was reduced. 1HNMR metabolomics analysis suggested that curcumin could regulate energy metabolism pathways. RT-qPCR and Western blot analysis found that the expression of myogenic factor myogenin was increased and the expression of myodegradation factor myostatin was decreased in the gastrocnemius; the level of ubiquitination and activation of the NF-κB pathway were also declined. Curcumin reduces ubiquitination, inflammation in skeletal muscle by regulating the NF-KB/UPS axis and improves muscle malignant metabolic phenotype and mitochondrial dysfunction, to alleviate muscle atrophy and loss of function in mice with breast cancer cachexia.