Ca2+-dependent protein phosphorylation in brush border membranes of rat kidney proximal tubules.
Ca2+-dependent protein phosphorylation was studied in isolated brush border membrane vesicles from rat kidney cortex. Phosphorylation of about 20 target proteins with gamma-32P-ATP was observed after opening the vesicles transiently by detergent treatment or by hypotonic shock indicating that phosphorylation takes place from the cytoplasmic side. Five of these polypeptides (Mr 12, 19, 29, 97 and 138 kD) showed increased phosphorylation in the presence of micromolar calcium. Addition of exogenous calmodulin did not enhance the calcium induced phosphorylation significantly, nor did trifluoperazine depress it, suggesting that Ca2+-calmodulin-dependent protein kinase is not involved in the studied Ca2+-induced phosphorylation. 12-O-tetradecanoylphorbol-13-acetate (TPA) minimized the Ca2+ dependency for the 12 and 97 kD polypeptides. Cytotoxin I inhibited the incorporation of phosphate into the 12 and 97 kD polypeptides in a dose-dependent manner. Excess phosphatidylserine could reverse this inhibition but stimulated also the phosphorylation of the 19 kD polypeptide. These findings suggest that at least the 12 and the 97 kD polypeptides are substrates for an endogenous protein kinase C. When studied under conditions where Ca2+ increases membrane phosphorylation, no effects could be observed on the kinetic parameters (Km and Vmax) of the sodium-dependent phosphate uptake.


Follicular lymphoma with progression to diffuse large B-cell lymphoma and concurrent CD5-negative mantle cell lymphoma-3 entities in a lymph node.
A 68-year-old woman with a history of follicular lymphoma had pathological findings of grade 3B follicular lymphoma, mantle cell lymphoma (MCL), and diffuse large B-cell lymphoma (DLBCL) identified in 1 lymph node. The DLBCL appeared to be a transformation of the follicular lymphoma. The nodules were diffusely and strongly positive for CD20, BCL6, and BCL2. CD43 highlighted smaller lymphocytes in a fraction of the nodules. BCL1 staining was variable with a mixture of nodular and mantle zone patterns. The diffuse areas showed weaker positivity for CD10, BCL2, and BCL6. CD3 and CD5 highlighted intermixed T cells. The Ki-67 proliferative index was overall estimated to be 60%. Fluorescent in situ hybridization performed on the lymph node was positive for CCND1/IGH. The patterns of BCL1 and BCL6 staining demonstrated 2 separate populations of neoplastic B lymphocytes.


A Ferrotoroidic Candidate with Well-Separated Spin Chains.
The search of novel quasi-1D materials is one of the important aspects in the field of material science. Toroidal moment, the order parameter of ferrotoroidic order, can be generated by a head-to-tail configuration of magnetic moment. It has been theoretically proposed that 1D dimerized and antiferromagnetic (AFM)-like spin chain hosts ferrotoroidicity and has the toroidal moment composed of only two antiparallel spins. Here, the authors report a ferrotoroidic candidate of Ba6 Cr2 S10 with such a theoretical model of spin chain. The structure consists of unique dimerized face-sharing CrS6 octahedral chains along the c axis. An AFM-like ordering at ≈10 K breaks both space- and time-reversal symmetries and the magnetic point group of mm'2'allows three ferroic orders in Ba6 Cr2 S10 : (anti)ferromagnetic, ferroelectric, and ferrotoroidic orders. Their investigation reveals that Ba6 Cr2 S10 is a rare ferrotoroid ic candidate with quasi 1D spin chain, which can be considered as a starting point for the further exploration of the physics and applications of ferrotoroidicity.


Pantothenate biosynthesis is critical for chronic infection by the neurotropic parasite Toxoplasma gondii.
Coenzyme A (CoA) is an essential molecule acting in metabolism, post-translational modification, and regulation of gene expression. While all organisms synthesize CoA, many, including humans, are unable to produce its precursor, pantothenate. Intriguingly, like most plants, fungi and bacteria, parasites of the coccidian subgroup of Apicomplexa, including the human pathogen Toxoplasma gondii, possess all the enzymes required for de novo synthesis of pantothenate. Here, the importance of CoA and pantothenate biosynthesis for the acute and chronic stages of T. gondii infection is dissected through genetic, biochemical and metabolomic approaches, revealing that CoA synthesis is essential for T. gondii tachyzoites, due to the parasite's inability to salvage CoA or intermediates of the pathway. In contrast, pantothenate synthesis is only partially active in T. gondii tachyzoites, making the parasite reliant on its uptake. However, pantothenate synthesis is crucial for the establishment of chronic infection, offering a promising target for intervention against the persistent stage of T. gondii.


The role of acrosin in sperm penetration through human cervical mucus.
Enzymes have been implicated in facilitating cervical mucus penetration by spermatozoa. One of these enzymes in the neutral proteinase acrosin, which is associated with the sperm acrosome. To determine the validity of this hypothesis, human spermatozoa were incubated with the following acrosin inhibitors: p-aminobenzamidine (AB), N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK), and p-nitropheyl-p'-guanidino benzoate (NPGB). An in vitro slide test system was developed which allowed inhibitor-treated and control spermatozoa to be evaluated against the same human cervical mucus sample. At inhibitor concentrations far exceeding those necessary for the inhibition of human acrosin, there was no effect on spermatozoal penetration into or through the mucus. These findings indicate that, in man, acrosin activity is neither necessary nor facilitory to sperm penetration of cervical mucus. Evidence is also presented that demonstrates the superiority of the newly developed double-interface slide test, especially for comparative purposes, over the tests currently in use.


D-penicillamine in pregnancy--to ban or not to ban?
The action of D-penicillamine on collagen can cause undesired side-effects in the treatment of cystinuria and Wilson's disease, it is on the other hand essential to the therapy of rheumatoid arthritis and scleroderma. Furthermore D-penicillamine can be potentially teratogenic, since it crosses the placental barrier. From the literature and our own observation of two pregnancies it is shown that among 87 pregnant women who received D-penicillamine 46 cases were treated during the whole period of pregnancy. Two infants from the latter group were found to have severe connective-tissue defects. We suggest that the dose of D-penicillamine in pregnant patients with cystinuria and Wilson's disease should be kept as low as possible. In the case of rheumatoid arthritis D-penicillamine should not be given during pregnancy.


Dilute performic acid--a versatile and easily to handle oxidant in general histology and histochemistry of structure bound sulphur compounds.
Performic acid normally leads to frequent and nearly inevitable detaching of commonly attached tissue sections, thus rendering such methods unrecommendable for histological and histochemical routine work. By the use of alcohol dilute solutions instead of the crude reagent, this problem can be managed easily. Special attention is drawn to the ambiguity of methods for tissue sulphur, based on the demonstration of oxidation induced strong basophilia.


COVID-19 among patients with orthopedic surgery: our experience from the Middle East.
We report our experiences with COVID-19 in one of the largest referral orthopedic centers in the Middle East and aimed to describe the epidemiology and clinical characteristics of these patients. During February 20 and April 20, 2020, patients who underwent orthopedic surgery and healthcare staff who were in contact with these patients were screened for COVID-19. To identify patients who were in the incubation period of COVID-19 during their hospital stay, all patients were tested again for COVID-19 4 weeks after discharge. Overall, 1244 patients underwent orthopedic surgery (1123 emergency and 121 elective) during the study period. Overall, 17 patients were diagnosed with COVID-19 during hospital admission and seven after discharge. Among the total 24 patients with COVID-19, 15 were (62.5%) males with a mean (SD) age of 47.0±1.6 years old. Emergency surgeries were performed in 20 (83.3%) patients, and elective surgery was done in the remaining 4 patients which included one case of posterior spinal fusion, spondylolisthesis, acromioclavicular joint dislocation, and one case of leg necrosis. A considerable number of infections occurred in patients with intertrochanteric fractures (n=7, 29.2%), followed by pelvic fractures (n=2, 8.3%), humerus fractures (n=2, 8.3%), and tibial plateau fractures (n=2, 8.3%). Fever (n=11, 45.8%) and cough (n=10, 37.5%) were the most common symptoms among patients. Laboratory examinations showed leukopenia in 2 patients (8.3%) and lymphopenia in 4 (16.7%) patients. One patient with a history of cancer died 2 weeks after discharge due to myocardial infarction. Among hospital staff, 26 individuals contracted COVID-19 during the study period, which included 13 (50%) males. Physicians were the most commonly infected group (n = 11), followed by operation room technicians (n = 5), nurses (n = 4), and paramedics (n = 4). Patients who undergo surgical treatment for orthopedic problems, particularly lower limb fractures with limited ambulation, are at a higher risk of acquiring COVID-19 infections, although they may not be at higher risks for death compared to the general population. Orthopedic surgeons in particular and other hospital staff who are in close contact with these patients must be adequately trained and given appropriate personal protective equipment during the COVID-19 outbreak.


Sialic acid content of infant saliva: comparison of breast fed with formula fed infants.
Sialic acid is found in especially high concentrations in brain gangliosides, and supplementary sialic acid is associated with increased learning behaviour in animals. It was hypothesised that breast fed infants may have higher concentrations of sialic acid in body fluids and tissues because human milk is a rich source of sialylated oligosaccharides, while formulas contain very little. The aim therefore was to compare the sialic acid content of saliva collected from full term infants who were either solely breast fed or formula fed until weaning at 3-5 months of age. Thirty three infants, 18 breast fed and 15 formula fed, were studied at a mean (SD) age of 5 (2) months. The breast fed infants, when compared with formula fed infants, were found to have almost two times more free sialic acid in saliva (mean (SE) 16.0 (2.7) v 8.2 (2.1) mg/l, p < 0.036) and nearly 50% more total sialic acid (47.3 (3.9) v 32.2 (4.4) mg/l, p < 0.014). The findings provide a preliminary indication that an exogenous source of sialic acids derived from human milk may contribute to higher concentrations of sialic acid in body fluids. There are important implications for the formulation of human milk substitutes.


Effects of preliminary normothermic flushing on hypothermic renal preservation.
Flushing of harvested kidneys with hypothermic perfusates is presumed to cause inappropriate circulation due to vasospasm. Effects of initial normothermic flushing prior to hypothermic flushing and cold preservation were tested in canine kidneys. Analysis of the perfusion characteristics revealed that a brief initial flushing with solutions at 37 degrees C. eliminates vasoconstriction and even facilitates subsequent hypothermic flushing. In amounts not exceeding 100 cc. it was not deleterious to transplant function. Larger volumes of normothermic flushing were proved to cause endothelial injury and resulted in poor transplant survival.


Primary structure of the carbohydrate chain of soybean agglutinin. A reinvestigation by high resolution 1H NMR spectroscopy.
In an earlier report (Lis, H., and Sharon, N. (1978) J. Biol. Chem. 253, 3468-3476) it was concluded that the plant glycoprotein soybean agglutinin contains two types of N-glycosidically linked carbohydrate units, both consisting of mannose and N-acetylgucosamine, in a molar ratio of 9:2. Both were described to have the same core structure, Man alpha (1 leads to 6)[Man alpha (1 leads to 3)]Man beta (1 leads to 4)GlcNAc beta (1 leads to 4)GlcNAc, but different patterns of branching. Here we present data showing that the asparagine-linked carbohydrate moiety of soybean agglutinin is homogeneous and possesses the following structure: (formula, see text) This conclusion is based on high resolution 1H NMR spectroscopy at 500 MHz of the isolated glycopeptide, and at 360 MHz of the oligosaccharide Man9GlcNAc obtained after digestion of the crude soybean agglutinin glycopeptide by endo-beta-N-acetylglucosaminidase H. The revised structure is identical in all respects with that of the high mannose N-glycosidic units of porcine thyroglobulin, of bovine lactotransferrin, and of the glycoprotein from Chinese hamster ovary cell membranes.


[Systemic manifestations and diseases related to positive serology for Yersinia pseudotuberculosis. Apropos of 9 observations].
A series of 9 cases of systemic affections associated with positive serology for yersinia pseudotuberculosis is reported. The clinical presentation of four cases was classical reactive arthritis with or without erythema nodosa; in two cases the patients had sacroileitis or Reiter's syndrome. The other three patients had unusual presentations: one Löfgren's syndrome, one cutaneous leukocytoclasis vasculitis with polyarthritis and glomerulonephritis, and one rhizomelic pseudopolyarthritis. Four patients (45 p. 100) had a positive HLA B27 antigen. In the absence of absolute bacteriological proof of cause and effect the authors discuss the relationship between these syndromes and Yersinia pseudotuberculosis infection. Although a Yersinia infection has not been formally established as the cause, there is some indirect evidence pointing to the reality of systemic Yersinia pseudotuberculosis infection.


Removal of bioaerosols by the combination of a photocatalytic filter and negative air ions.
This study focused on the investigation of the effectiveness of negative air ionization (NAI), photocatalytic oxidation (PCO), and the combination of NAI and PCO on the removal of aerosolized Escherichia coli, Candida famata, and λ vir phage under different relative humidity. The experiments were conducted with a stainless steel reactor equipped with a negative air ion generator, a photocatalytic filter, and two ultraviolet lamps with 365 nm wavelength. The removal efficiency ( η ) , defined as one minus the ratio of the outlet concentration to the inlet concentration of the appropriate bioaerosol, was used to evaluate the effectiveness of the removal methods. The combination of NAI and PCO was the most efficient removal method for aerosolized E. coli ( η = 0.304 ± 0.06 - 0.364 ± 0.008 ) , C. famata ( η = 0.433 ± 0.08 - 0.598 ± 0.047 ) , and λ vir phage ( η = 0.689 ± 0.02 - 0.903 ± 0.06 ) . In this removal method, the contributions of NAI were higher than those of PCO for the removal of E. coli and C. famata; for the removal of λ virus phage the contributions of NAI and PCO were comparable NAI was the least efficient removal method for bioaerosols, and the removal efficiencies are: η = 0.175 ± 0.04 - 0.245 ± 0.03 for E. coli; η = 0.216 ± 0.007 - 0.297 ± 0.044 for C. famata; and η = 0.299 ± 0.12 - 0.384 ± 0.02 for λ vir phage.


Characteristics of basal taurine release in the rat striatum measured by microdialysis.
Taurine is a sulfur-containing amino acid thought to be an osmoregulator, neurotransmitter or neuromodulator in the brain. Our objective was to establish how much taurine is released in the striatum and examine the mechanisms controlling extracellular taurine concentrations under resting conditions. The experiments were made on rats by microdialysis in vivo. Changes in taurine were compared with those in glutamate, glycine and the non-neuroactive amino acid threonine. Using the zero net flux approach we showed the extracellular concentration of taurine to be 25.2 +/- 5.1 muM. Glutamate was increased by tetrodotoxin and decreased by Ca2+ omission, glycine and threonine were not affected and both treatments increased extracellular taurine. The basal taurine release was increased by the taurine transport inhibitor guanidinoethanesulfonate and reduced by the anion channel blocker 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid.


[Bioelectric activity of the myocardium and central hemodynamics in patients with vibration disease].
Examination in occupational clinic covered 60 male sufferers from vibration disease caused by local vibration (28 patients with I and I-II stages, 26 with II stage, 6 with residual signs). Nearly 36% of examinees demonstrated moderate disorders of bioelectric myocardial activity in left ventricle. Hyperkinetic type of central hemodynamics was diagnosed in 75% of the subjects, hypokinetic one-in 8.3%, eukinetic one-in 16.7%. The hyperkinetic type may promote arterial hypertension during vibration disease.


Association between Cerebral Performance Category, Modified Rankin Scale, and discharge disposition after cardiac arrest.
Cerebral Performance Category (CPC), Modified Rankin Scale (mRS) and discharge disposition are commonly used to determine outcomes following cardiac arrest. This study tested the association between these outcome measures. Retrospective chart review of subjects who survived to hospital discharge between 1/1/2006 and 12/31/2009 was conducted. Charts were reviewed for outcomes (CPC, mRS, and discharge disposition). Discharge disposition was classified in 6 categories: home with no services, home with home healthcare, acute rehabilitation facility, skilled nursing facility, long term acute care facility, and hospice. Intra-and inter-rater reliabilities were calculated for outcome measures. Rates of "good outcome" (defined as a CPC of 1-2, mRS of 0-3, or discharge disposition to home or acute rehabilitation facility) were also determined. Kendall's tau correlation coefficients explored relationships among measures. A total of 211 charts were reviewed. Mean age was 60 years (SD 16), the majority (75%) were white males, in- and out-of hospital cardiac arrests were equally prevalent, and ventricular dysrhythmia was most common (N=109, 52%). Half of the subjects were comatose following resuscitation and 75 (35%) received therapeutic hypothermia. Inter-rater percentage agreement for CPC and mRS abstraction was 95.24% (kappa 0.89, p<0.001) and 95.24% (kappa 0.90, p<0.001) respectively. "Good outcomes" were found in 44 subjects (20%) using the CPC definition, 47 subjects (22%) using the mRS definition, and 129 subjects (61%) subjects using discharge disposition definition. There was fair relationship between the CPC and mRS (tau 0.43) and poor relationships between CPC and discharge disposition (tau 0.23) and between mRS and discharge disposition (tau 0.25). Determination of the CPC, mRS and discharge disposition at hospital discharge is reliable from chart review. These instruments provide widely differing estimates of "good outcome". Agreement between these measures ranges from poor to fair. A more nuanced outcome measure designed for the post-cardiac arrest population is needed.


Associations of Life's Simple 7 with cerebral white matter hyperintensities and microstructural integrity: UK Biobank cohort study.
The American Heart Association Life's Simple 7 (LS7) metric was used to define optimal cardiovascular and brain health, but the associations with macrostructural hyperintensities and microstructural white matter damage are unclear. The objective was to determine the association of LS7 ideal cardiovascular health factors with macrostructural and microstructural integrity. A total of 37,140 participants with available LS7 and imaging data from UK Biobank were included in this study. Linear associations were implemented to examine the associations of LS7 score and subscores with white matter hyperintensity load (WMH) (WMH volume normalized by total white matter volume and logit-transformed) and diffusion imaging indices (fractional anisotropy [FA], mean diffusivity, orientation dispersion index [OD], intracellular volume fraction, isotropic volume fraction [ISOVF]). In individuals (mean age 54.76 years; 19,697 females, 52.4%), higher LS7 score and subscores were strongly associated with lower WMH and microstructural white matter injury, including OD, ISOVF, FA. Both interaction analyses and stratified analyses of LS7 score and subscores with age and sex showed a strong association with microstructural damage markers, with remarkable age and sex differences. The association of OD was pronounced in females and populations younger than 50 years and FA, mean diffusivity and ISOVF were pronounced in males and populations older than 50 years. These findings suggest that healthier LS7 profiles are associated with better profiles of both macrostructural and microstructural markers of brain health, and indicate that ideal cardiovascular health is associated with improved brain health.


Fine-scale variation in microhabitat conditions influences physiology and metabolism in an Antarctic insect.
Microhabitats with distinct biotic and abiotic properties exist within landscapes, and this microhabitat variation can have dramatic impacts on the phenology and physiology of the organisms occupying them. The Antarctic midge Belgica antarctica inhabits diverse microhabitats along the Western Antarctic Peninsula that vary in macrophyte composition, hygric qualities, nutrient input, and thermal patterns. Here, we compare seasonal physiological changes in five populations of B. antarctica living in close proximity but in different microhabitats in the vicinity of Palmer Station, Antarctica. Thermal regimes among our sample locations differed in both mean temperature and thermal stability. Between the warmest and coldest sites, seasonal mean temperatures differed by 2.6˚C and degree day accumulations above freezing differed by a factor of 1.7. Larval metabolic and growth rates varied among the sites, and adult emergence occurred at different times. Distinct microhabitats also corresponded with differences in body composition, as lipid and carbohydrate content of larvae differed across sites. Further, seasonal changes in carbohydrate and protein content were dependent on site, indicating fine-scale variation in the biochemical composition of larvae as they prepare for winter. Together, these results demonstrate that variation in microhabitat properties influences the ontogeny, phenology, physiology, and biochemical makeup of midge populations living in close proximity. These results have implications for predicting responses of Antarctic ecosystems to environmental change.


The utility of imputation for molecular mismatch analysis in solid organ transplantation.
HLA genotyping has undergone a rapid progression in resolution since the development of DNA-based typing methods. Despite the advent of high-resolution next-generation sequencing, the bulk of solid organ genotyping is performed at intermediate resolution, which provides multiple possible two-field results for each classical HLA loci. As a result, several methodologies have been developed to impute the most likely allele-level (two-field) HLA genotype for the purposes of donor-recipient compatibility analysis. The advent of molecular mismatch analysis, however, has placed a new emphasis on the accuracy of imputation. While seminal molecular mismatch studies have relied on the imputation of intermediate resolution genotyping, several recent studies have performed analysis showing that imputation generates inaccuracies in epitope identification. While the clinical impact of these errors is not clear, it is important that these concerns do not preclude future progress in understanding the utility of molecular mismatch analysis in transplantation. In the future, advances in genotyping methods will result in routine two-field resolution that will abrogate these concerns. In the meantime, however, studies are needed in order to address the role of molecular mismatch in diverse patient populations and to carefully address the potential of molecular mismatch analysis in the context of imputation.


Researchers, Patients, and Stakeholders Evaluating Comparative-Effectiveness Research: A Mixed-Methods Study of the PCORI Reviewer Experience.
The Patient-Centered Outcomes Research Institute (PCORI) includes patients and stakeholders alongside scientists in reviewing research applications using unique review criteria including patient-centeredness and patient and/or stakeholder engagement. To support extension of this unique collaborative model to other funders, information from the reviewers on the review process is needed to understand how scientists and nonscientists evaluate research proposals together. Thus, this study aimed to describe reviewers' perspectives of the interactions during the in-person review panel; to examine the value and challenges of including scientists, patients, and stakeholders together; and to understand the perceived importance of PCORI's review criteria. This study utilized anonymous, cross-sectional surveys (N = 925 respondents from 5 funding cycles: 470 scientists, 217 patients, 238 stakeholders; survey completion rates by cycle: 70-89%) and group interviews (N = 18). Reviewers of all types describe PCORI Merit Review as respectful, balanced, and one of reciprocal influence among different reviewer types. Reviewers indicate strong support and value of input from all reviewer types, receptivity to input from others, and the panel chair's incorporation of all views. Patients and stakeholders provide real-world perspectives on importance to patients, research partnership plans, and study feasibility. Challenges included concerns about a lack of technical expertise of patient/stakeholder reviewers and about scientists dominating conversations. The most important criterion for assigning final review scores was technical merit-either alone or in conjunction with patient-centeredness or patient/ stakeholder engagement. PCORI Merit Reviewers' self-reports indicate that the perspectives of different reviewer types are influential in panel discussions and Merit Review outcomes.


Direct encapsulation of AIE-active dye with β cyclodextrin terminated polymers: Self-assembly and biological imaging.
Aggregation-induced emission (AIE) phenomenon has attracted great attention recently and been extensively explored for biomedical applications. Nevertheless, the direct utilization of AIE-active dyes for biomedical applications has demonstrated to be enormous challenge owing to the hydrophobic nature of these AIE-active dyes. In this work, we reported the fabrication of amphiphilic AIE-active copolymers through the specific host-guest interaction between β cyclodextrin (β-CD) and an adamantine terminating tetraphenylethene derivative (TPE-Ad). In this construction system, β-CD was acted as the bridge to link TPE-Ad with PEG. The TPE-β-CD-PEG copolymers were characterized by various equipments in detail. Cytocompatibility and cell uptake behavior of TPE-β-CD-PEG were also examined to evaluate their biomedical application potential. Results demonstrated that TPE-β-CD-PEG copolymers were prone to self-assemble into luminescent nanoparticles, which exhibited high water dispersity, AIE feature and excellent biocompatibility. These features endowed TPE-β-CD-PEG great potential for biomedical applications.


Progression and regression of sleep-disordered breathing with changes in weight: the Sleep Heart Health Study.
The relationship of weight changes to the incidence, progression, and remission of sleep-disordered breathing (SDB) is not well defined. This study aims to determine the relationship between change in weight and progression or remission of SDB by polysomnography. We performed a longitudinal cohort study of the cardiovascular consequences of sleep apnea in diverse US communities. Sleep apnea and polysomnographic indicators of SDB were assessed 5 years apart. A total of 2968 men and women (mean age, 62 years) participated in the study. Men were more likely to have an increase in Respiratory Disturbance Index (RDI) with a given increase in weight than were women, and this was not explained by differences in starting weight, waist circumference, age, or ethnicity. In a linear regression analysis, both men and women had a greater increase in RDI with weight gain than a decrease in RDI with weight loss. In a categorical analysis of larger degrees of change, this sex difference was also evident. Associations were similar in diverse ethnic groups. However, SDB progressed over time, even in those with stable weight. Modest changes in weight were related to an increase or decrease in SDB, and this association was stronger in men than in women.


Association of deranged cerebrovascular reactivity with brain injury following cardiac arrest: a post-hoc analysis of the COMACARE trial.
Impaired cerebrovascular reactivity (CVR) is one feature of post cardiac arrest encephalopathy. We studied the incidence and features of CVR by near infrared spectroscopy (NIRS) and associations with outcome and biomarkers of brain injury. A post-hoc analysis of 120 comatose OHCA patients continuously monitored with NIRS and randomised to low- or high-normal oxygen, carbon dioxide and mean arterial blood pressure (MAP) targets for 48 h. The tissue oximetry index (TOx) generated by the moving correlation coefficient between cerebral tissue oxygenation measured by NIRS and MAP was used as a dynamic index of CVR with TOx > 0 indicating impaired reactivity and TOx > 0.3 used to delineate the lower and upper MAP bounds for disrupted CVR. TOx was analysed in the 0-12, 12-24, 24-48 h time-periods and integrated over 0-48 h. The primary outcome was the association between TOx and six-month functional outcome dichotomised by the cerebral performance category (CPC1-2 good vs. 3-5 poor). Secondary outcomes included associations with MAP bounds for CVR and biomarkers of brain injury. In 108 patients with sufficient data to calculate TOx, 76 patients (70%) had impaired CVR and among these, chronic hypertension was more common (58% vs. 31%, p = 0.002). Integrated TOx for 0-48 h was higher in patients with poor outcome than in patients with good outcome (0.89 95% CI [- 1.17 to 2.94] vs. - 2.71 95% CI [- 4.16 to - 1.26], p = 0.05). Patients with poor outcomes had a decreased upper MAP bound of CVR over time (p = 0.001), including the high-normal oxygen (p = 0.002), carbon dioxide (p = 0.012) and MAP (p = 0.001) groups. The MAP range of maintained CVR was narrower in all time intervals and intervention groups (p < 0.05). NfL concentrations were higher in patients with impaired CVR compared to those with intact CVR (43 IQR [15-650] vs 20 IQR [13-199] pg/ml, p = 0.042). Impaired CVR over 48 h was more common in patients with chronic hypertension and associated with poor outcome. Decreased upper MAP bound and a narrower MAP range for maintained CVR were associated with poor outcome and more severe brain injury assessed with NfL. Trial registration ClinicalTrials.gov, NCT02698917 .


A general approach to the synthesis of 5-S-functionalized pyrimidine nucleosides and their analogues.
A general and efficient approach was developed for the introduction of S-functionality at the C-5 position of cytosine and uracil nucleosides and their analogues. The key step is a palladium-catalyzed C-S coupling of the corresponding 5-bromo nucleoside derivative and alkyl thiol. The butyl 3-mercaptopropionate coupling products were further converted to the corresponding disulphides, the stable precursors of 5-mercaptopyrimidine nucleosides.


Measurements of wall shear stress and aortic pulse wave velocity in swine with familial hypercholesterolemia.
To assess measurements of pulse wave velocity (PWV) and wall shear stress (WSS) in a swine model of atherosclerosis. Nine familial hypercholesterolemic (FH) swine with angioplasty balloon catheter-induced atherosclerotic lesions to the abdominal aorta (injured group) and 10 uninjured FH swine were evaluated with a 4D phase contrast (PC) magnetic resonance imaging (MRI) acquisition, as well as with radial and Cartesian 2D PC acquisitions, on a 3T MR scanner. PWV values were computed from the 2D and 4D PC techniques, compared between the injured and uninjured swine, and validated against reference standard pressure probe-based PWV measurements. WSS values were also computed from the 4D PC MRI technique and compared between injured and uninjured groups. PWV values were significantly greater in the injured than in the uninjured groups with the 4D PC MRI technique (P = 0.03) and pressure probes (P = 0.02). No significant differences were found in PWV between groups using the 2D PC techniques (P = 0.75-0.83). No significant differences were found for WSS values between the injured and uninjured groups. The 4D PC MRI technique provides a promising means of evaluating PWV and WSS in a swine model of atherosclerosis, providing a potential platform for developing the technique for the early detection of atherosclerosis.


Disseminated Intravascular Coagulation Score Predicts Mortality in Patients with Liver Disease and Low Fibrinogen Level.
Alongside its original diagnostic intention, the International Society on Thrombosis and Haemostasis' (ISTH) disseminated intravascular coagulation (DIC) score predicts mortality in various patient groups. We investigated whether coagulopathy quantified by the DIC score can predict 30-day mortality in patients with liver disease and low fibrinogen levels. We retrospectively analyzed all patients admitted to the Vienna General Hospital between 2003 and 2014 with a fibrinogen level of <150 mg/dL, a history of liver disease, and ≥2 pathological DIC parameters. We used a Cox regression and receiver operating characteristic analysis to assess the predictive value of the ISTH DIC score in its original (DIC-2001) and revised form (DIC-2018). A total of 1,333 patients were screened, and 388 of these patients (38% female, median age: 58 years, interquartile range: 48-66 years) were analyzed. The DIC-2001 (hazard ratio [HR]: 2.08, 95% confidence interval [CI]: 1.78-2.59, p < 0.001) and DIC-2018 (HR: 1.73, 95% CI: 1.51-2.05, p < 0.001) predicted 30-day mortality. The results remained robust in several sensitivity analyses. The ISTH DIC-2001 and DIC-2018 scores predicted 30-day mortality in patients with liver disease and low fibrinogen levels. The DIC score deserves further investigation in this population as it likely reflects different dimensions of the underlying disease.


Gene therapy with tumor-specific promoter mediated suicide gene plus IL-12 gene enhanced tumor inhibition and prolonged host survival in a murine model of Lewis lung carcinoma.
Gene therapy is a promising therapeutic approach for cancer. Targeted expression of desired therapeutic proteins within the tumor is the best approach to reduce toxicity and improve survival. This study is to establish a more effective and less toxic gene therapy of cancer. Combined gene therapy strategy with recombinant adenovirus expressing horseradish peroxidase (HRP) mediated by human telomerase reverse transcriptase (hTERT) promoter (AdhTERTHRP) and murine interleukin-12 (mIL-12) under the control of Cytomegalovirus (CMV) promoter (AdCMVmIL-12) was developed and evaluated against Lewis lung carcinoma (LLC) both in vivo and in vitro. The mechanism of action and systemic toxicities were also investigated. The combination of AdhTERTHRP/indole-3-acetic acid (IAA) treatment and AdCMVmIL-12 resulted in significant tumor growth inhibition and survival improvement compared with AdhTERTHRP/IAA alone (tumor volume, 427.4 ± 48.7 mm3 vs 581.9 ± 46.9 mm3, p = 0.005 on day 15; median overall survival (OS), 51 d vs 33 d) or AdCMVmIL-12 alone (tumor volume, 362.2 ± 33.8 mm3 vs 494.4 ± 70.2 mm3, p = 0.046 on day 12; median OS, 51 d vs 36 d). The combination treatment stimulated more CD4+ and CD8+ T lymphocyte infiltration in tumors, compared with either AdCMVmIL-12 alone (1.3-fold increase for CD4+ T cells and 1.2-fold increase for CD8+ T cells, P < 0.01) or AdhTERTHRP alone (2.1-fold increase for CD4+ T cells and 2.2-fold increase for CD8+ T cells, P < 0.01). The apoptotic cells in combination group were significantly increased in comparison with AdCMVmIL-12 alone group (2.8-fold increase, P < 0.01) or AdhTERTHRP alone group (1.6-fold increase, P < 0.01). No significant systematic toxicities were observed. Combination gene therapy with AdhTERTHRP/IAA and AdCMVmIL-12 could significantly inhibit tumor growth and improve host survival in LLC model, without significant systemic adverse effects.


Analysis of late-blight disease resistance and freezing tolerance in transgenic potato plants expressing sense and antisense genes for an osmotin-like protein.
The expression patterns of plant defense genes encoding osmotin and osmotin-like proteins imply a dual function in osmotic stress and plant pathogen defense. We have produced transgenic potato (Solanum commersonii Dun.) plants constitutively expressing sense or antisense RNAs from chimeric gene constructs consisting of the cauliflower mosaic virus 35S promoter and a cDNA (pA13) for an osmotin-like protein. Transgenic potato plants expressing high levels of the pA13 osmotin-like protein showed an increased tolerance to the late-blight fungus Phytophthora infestans at various phases of infection, with a greater resistance at an early phase of fungal infection. There was a decrease in the accumulation of osmotin-like mRNAs and proteins when antisense transformants were challenged by fungal infection, although the antisense transformants did not exhibit any alterations in disease susceptibility. Expression of pA13 sense and antisense RNAs had no effect on the development of freezing tolerance in transgenic plants when assayed under a variety of conditions including treatments with abscisic acid or low temperature. These results provide evidence of antifungal activity for a potato osmotin-like protein against the fungus P. infestans, but do not indicate that pA13 osmotin-like protein is a major determinant of freezing tolerance.


Biochemical characterization of an organic solvent-tolerant glycosyltransferase from Bacillus licheniformis PI15 with potential application for raspberry ketone glycoside production.
Raspberry ketone is a primary aroma component of the red raspberry. The glycosylation of this compound is a potential approach used to improve its pharmaceutical properties. In this work, raspberry ketone glycosides are produced in bacteria for the first time. Bacillus licheniformis PI15, an organic solvent-tolerant glycosyltransferase-producing strain, was isolated from chemically polluted soil. The cloning and heterologous expression of a glycosyltransferase, which was designated PI-GT1, in Escherichia coli BL21 resulted in the expression of an active and soluble protein that accounted for 15% of the total cell protein content. Purified PI-GT1 was highly active and stable over a broad pH range (6.0-10.0) and showed excellent pH stability. PI-GT1 maintained almost 60% of its maximal activity after 3 H of incubation at 20-40 °C and demonstrated optimal activity at 30 °C. Additionally, PI-GT1 displayed high stability and activity in the presence of hydrophilic solvents with log P ≤ -0.2 and retained more than 80% of its activity after 3 H of treatment. Supplementation with 10% DMSO markedly improved the glycosylation of raspberry ketone, resulting in a value 26 times higher than that in aqueous solution. The organic solvent-tolerant PI-GT1 may have potential uses in industrial chemical and pharmaceutical synthesis applications.


The crippling effects of a cyberattack at an academic level 1 trauma center: An orthopedic perspective.
Malicious cyberattacks are increasing in frequency and severity with healthcare institutions spending an average of over 10 million dollars to resolve the consequences of healthcare data breaches. This cost does not include the effect of a downtime event should a healthcare system electronic medical record (EMR) lose functionality. An Academic Level 1 trauma center suffered a cyberattack resulting in a total EMR downtime of 25 days. Orthopedic operative time was used as a surrogate for OR functionality during the event and a framework with specific examples is presented to promote rapid adaptation during downtime events. Operative time losses were identified by calculating a running average of weekday total in room operative time during a total downtime event secondary to a cyberattack. This data was compared to week-of-the-year matched data from the year prior and the year after the attack. A framework for creating adaptations to a total downtime event was created by repeatedly interviewing different provider groups and identifying how they adjusted care to the challenges faced. Total weekday in room operative time during the attack decreased by 53.4% ± 12.2% and 53.2% ± 14.9% when comparing the matched period one year prior and one year after, respectively. Immediate challenges to patient care were identified by small groups of highly motivated individuals, with self-assigned agile teams formed. These teams sequenced system processes, identified failure points, and created real-time solutions. A frequently updated EMR backup mirror and hospital disaster insurance were crucial for mitigating the impact of the cyberattack. Cyberattacks are expensive and their downstream effects, including downtime events, can be crippling. Agile team formation, process sequencing, and understanding EMR backup times are tactics used to combat the challenges of a prolonged total downtime event. Level III retrospective cohort.


Inhibition of β1 integrin induces its association with MT1-MMP and decreases MT1-MMP internalization and cellular invasiveness.
Integrin signaling plays a fundamental role in the establishment of focal adhesions and the subsequent formation of invadopodia in malignant cancer cells. Invadopodia facilitate localized adhesion and degradation of the extracellular matrix (ECM), which promote tumour cell invasion and metastasis. Degradation of ECM components is often driven by membrane type-1 matrix metalloproteinase (MT1-MMP), and we have recently shown that regulation of enzyme internalization is dependent on signaling downstream of β1 integrin. Phosphorylation of the cytoplasmic tail of MT1-MMP is required for its internalization and delivery to Rab5-marked early endosomes, where it is then able to be recycled to new sites of invadopodia formation and promote invasion. Here we found that inhibition of β1 integrin, using the antibody AIIB2, inhibited the internalization and recycling of MT1-MMP that is necessary to support long-term cellular invasion. MT1-MMP and β1 integrin were sequestered at the cell surface when β1-integrin was inhibited, and their association under these conditions was detected using immunoprecipitation and mass spectrometry analyses. Sequestration of β1 integrin and MT1-MMP at the cell surface resulted in the formation of large invadopodia and local ECM degradation; however, the impaired internalization and recycling of MT1-MMP and β1 integrin ultimately led to a loss of invasive behaviour.


Closure of large myelomeningocoele with Ramirez technique.
The authors present their 7-year experience in operative closure of large open myelomeningocoele with a technique introduced by Ramirez et al. in 1986. It consists of medial advancement of bilateral bipedicled musculocutanous flap based on the latissimus dorsi and maximus gluteus without any relaxing incisions or skin grafting. Thirty-five babies were operated on according to this technique on first day of life. In 16 cases, large thoracic or thoracolumbosacral defect was noted, while in further 19 newborns, lumbosacral or lumbar defect occurred. The dimensions of the entire defect ranged from 4.8x5 to 8x10.8 cm. In seven babies, the affected area had more than 50 cm(2). In all patients, three-layer tension-free closure of the dorsal wound was performed in one stage. The mean operative time was 115 min (90-195 min). All but two wounds healed uneventfully. In two babies with wide lumbosacral defects, minor skin necrosis with wound dehiscence was noted which subsequently required conservative treatment. Follow-up period ranges from 6 months to 7 years. In none of the 32 children was any late complication of the reconstructive procedure noted. Ramirez technique is a very reliable method of closure of myelomeningocoele regardless of the size of the defect and its topography. It may find a wide application in babies affected by neural tube defect.


Preparation, characterization, dissolution, and permeation of flibanserin - 2-HP-β-cyclodextrin inclusion complexes.
Flibanserin (FLB), an antiserotonin drug, is used to treat women with hypoactive sexual appetite disorder. FLB shows low bioavailability (~33%) probably due to its low water solubility. The current study investigated the impact of hydroxypropyl-β-cyclodextrin (HP-β-CD) and sodium lauryl sulfate (SLS) on the dissolution and permeation of FLB. HP-β-CD-FLB inclusion complexes were prepared using physical mixing and kneading at 1:1 and 1:2 M ratios and characterized using differential scanning calorimetry, Fourier transform infrared spectroscopy, and powder X-ray diffractometry. The dissolution and permeation of the complexes through a cellophane membrane were performed in, 0.1, 0.3 and 0.5% SLS in phosphate buffer (pH 6.8). Derived from the slope of the linear phase solubility diagram, the apparent stability constant (K 1:1) was 372.54 M-1. Kneading changed the crystalline form of FLB to an amorphous appearance characterized by minimal crystalline peaks, indicating successful inclusion complex formation. In addition, the HP-β-CD-FLB inclusion complexes showed twofold increased dissolution efficiency at 6 h. The cumulative FLB amount permeated at 6 h increased from 14.1% to 21.88% and 34.56% in the presence of 0.1% and 0.3% of SLS, respectively. However, increasing SLS to 0.5% did not show an increase in FLB permeation. Therefore, the HP-β-CD-FLB inclusion complex has an improved dissolution rate compared to FLB alone. The presence of SLS in the dissolution medium increases the dissolution rate of pure FLB and its complex with HP-β-CD. kneaded 1:1 complex was formulated bioadhesive buccal tablets and showed enhanced drug release.


DER revises waste management rules.
A number of improvements were made in new regulations designed to help physicians manage their infectious and chemotherapeutic wastes more easily. This article summarizes those improvements.


The role of perceptual, cognitive, and motor abilities in street-crossing decisions of young and older pedestrians.
The present experiment investigated the role of perceptual, cognitive, and motor abilities in street-crossing behaviour with ageing. Previous research has shown that older pedestrians make many unsafe crossing decisions when cars are approaching at high speeds, and miss many crossing opportunities when car speeds are low. The older subjects seem to ignore information about the speed of the approaching cars and to preferentially use simplifying heuristics based on vehicle distance. The objective of the present study was to better understand the underlying age-related changes that lead to these behaviours, with a specific focus on perceptual factors. Twenty young (age 20-30), 21 younger-old (age 61-71), and 19 older-old (age 72-83) participants took part in the experiment. All participants individually carried out a simulated street-crossing task and took a battery of functional tests assessing perceptual, cognitive, and motor abilities. In line with earlier findings, the seniors made a greater number of incorrect crossing decisions, with many risky decisions when the vehicle was approaching at a high speed and many missed opportunities at a low speed. Correlation and regression analyses pointed out several functional performance measures as predictors of the way the pedestrians took or did not take information about vehicle speed into account in their decisions. Processing speed and visual attention abilities were shown to play the most important role in explaining the variance in incorrect decisions: these abilities allowed participants to focus their attention on the relevant speed information and to make timely, correct decisions. Time-to-arrival estimates, which informed the pedestrians about the time available for crossing, were found to be the second most predictive factor. Walking speed, by way of which the pedestrians adapted their crossing pace to the perceived available time, also came into play. Inhibition abilities ended up as the last functional predictor; they allowed the pedestrians to ignore irrelevant information and inhibit automatic but unsuitable responses. The present study provided a multidimensional explanation of increased gap-selection difficulties with ageing, including a combination of perceptual, cognitive, as well as physical performance declines with increasing age. The findings have implications for improving older pedestrians' safety in terms of speed limits, road design, and training.


Prevalence of plantar ulcer and its risk factors in leprosy: a systematic review and meta-analysis.
Plantar ulcers are a leading complication of leprosy that requires frequent visits to hospital and is associated with stigma. The extent of burden of ulcers in leprosy and its risk factors are scant impeding the development of targeted interventions to prevent and promote healing of ulcers. The aim of this review is to generate evidence on the prevalence of plantar ulcer and its risk factors in leprosy. Databases (Medline, Embase, Web of Science, CINAHL, BVS), conference abstracts and reference lists were searched for eligible studies. Studies were included that reported a point prevalence of plantar ulcer and/or its "risk factors" associated with development of ulcers (either causatively or predictively), including individual level, disease related and bio-mechanical factors. We followed PRISMA guidelines for this review. Random-effects meta-analysis was undertaken to estimate the pooled point prevalence of ulcers. Reported risk factors in included studies were narratively synthesised. This review is registered in PROSPERO: CRD42022316726. Overall, 15 studies (8 for prevalence of ulcer and 7 for risk factors) met the inclusion criteria. The pooled point prevalence of ulcer was 34% (95% CIs: 21%, 46%) and 7% (95% CIs: 4%, 11%) among those with foot anaesthesia and among all people affected by leprosy, respectively. Risk factors for developing ulcers included: unable to feel 10 g of monofilament on sensory testing, pronated/hyper-pronated foot, foot with peak plantar pressure, foot with severe deformities, and those with lower education and the unemployed. The prevalence of plantar ulceration in leprosy is as high as 34% among those with loss of sensation in the feet. However, the incidence and recurrence rates of ulceration are least reported. The inability to feel 10 g of monofilament appears to be a strong predictor of those at risk of developing ulcers. However, there is a paucity of evidence on identifying those at risk of developing plantar ulcers in leprosy. Prospective studies are needed to estimate the incidence of ulcers. Identifying individuals at risk of ulcers will help design targeted interventions to minimize risk factors, prevent ulcers and promote ulcer healing.


Single-copy T-DNA insertions in Arabidopsis are the predominant form of integration in root-derived transgenics, whereas multiple insertions are found in leaf discs.
Different patterns of T-DNA integration in Arabidopsis were obtained that depended on whether a root or a leaf-disc transformation method was used. An examination of 82 individual transgenic Arabidopsis plants, derived from 15 independent Agrobacterium-mediated transformations in which different cointegrate and binary constructs were used, indicated that the transformation method had a significant influence on the type and copy number of T-DNA integration events. Southern hybridizations showed that most of the transgenic plants produced by a leaf-disc method contained multiple T-DNA insertions (89%), the majority of which were organized as right-border inverted repeat structures (58%). In contrast, a root transformation method mostly resulted in single T-DNA insertions (64%), with fewer right-border inverted repeats (38%). The transformation vectors, including cointegrate and binary types, and the plant selectable markers, hygromycin phosphotransferase and dihydrofolate reductase, did not appear to influence the T-DNA integration patterns.


N5-phosphonoacetyl-L-ornithine (PALO): a convenient synthesis and investigation of influence on regulation of amino acid biosynthetic genes in Saccharomyces cerevisiae.
A scalable four-step synthesis of the ornithine transcarbamylase inhibitor N(5)-phosphonoacetyl-l-ornithine (PALO) is achieved through boroxazolidinone protection of ornithine. Investigations in the model organism Saccharomyces cerevisiae found that, in contrast to a previous report, PALO did not influence growth rate or expression of genes involved in arginine metabolism.


Protective effect of hop beta-acids on microbial degradation of thick juice during storage.
This study assessed the value of a commercial alkaline solution of hop beta-acids (HBA) for prevention of microbial degradation of thick juice, a concentrated intermediate product in the production of beet sugar. The antimicrobial effect of different concentrations of HBA against microbial degradation of thick juice was tested in a pilot-scale storage experiment. Chemical, biochemical and microbial parameters were monitored during thick juice storage. Thick juice degradation, indicated as a decrease in pH, was generally accompanied by an increase in the count of fastidious bacteria (FB) on Columbia Agar with Sheep Blood (CAwSB), which were mainly identified as Tetragenococcus halophilus. Addition of HBA delayed juice acidification and the development of FB in a concentration-dependent manner. The susceptibility of FB to HBA was determined by plating degraded thick juice (FB > 10(5) CFU ml(-1) on CAwSB plates with different concentrations of HBA (0-160 ppm). None of the HBA concentrations tested reduced the number of FB colonies formed, but increasing HBA concentrations extended the lag time of colony formation. HBA produce no measurable bactericidal effect, but retard the development of FB in thick juice. Moreover, HBA do not prevent the thick juice from deteriorating, but significantly delay its degradation. These results indicate that adding a commercially available HBA formulation can prolong the storage life of thick juice in the sugar industry, although degradation cannot be eliminated. Future research will focus on the detailed characterization of FB consistently isolated from degraded thick juice and on determining their role in thick juice degradation.


Genetic and phenotypic heterogeneity of multiple lentigines and precise diagnosis in four Chinese families with multiple lentigines.
Lentigines are well-defined, small, brown macules resulting from the accumulation of melanin content in the basement membrane zone with an increase in the number of melanocytes. Hereditary multiple lentigines (ML) can be associated with multiple genes and are not commonly encountered in clinical practice. Patients can solely have skin involvement or present with multisystemic deformative phenotypes. This study aimed to describe four unrelated Chinese families presenting with ML as their first visit symptom. We performed whole-exome sequencing (WES) and Sanger sequencing on all patients and immediate family members for precise molecular diagnosis. Two novel variants c.1548 T > A (p.Ser516Arg) and c.1811C > A (p.Thr604Lys) in SASH1, and two recurrent variants c.1403C > T (p.Thr468Met) and c.1493G > T (p.Arg498Leu) in PTPN11, were identified in these four families. We also summarized the genes associated with ML and differential diagnosis of pigment abnormality. We suggested that the molecular diagnosis of ML should be emphasized because it can help in the clinical differential diagnosis and further genetic counseling and prognosis.


Augmented therapeutic efficacy of 5-fluorouracil in conjunction with lantibiotic nisin against skin cancer.
Chemotherapy, a gold standard for treating most of the cancers, involves drastic side-effects and multidrug resistance. An attractive alternative is development of combination therapy employing antimicrobial peptides with chemotherapeutic drugs. In vivo studies: Anti-cancer therapeutic efficacy of 5-fluororuacil (5-FU) in conjunction with nisin (50 mg/kg body weight) was evaluated against murine skin cancer, in terms of tumor biostatistics, histopathology, electron microscopy, infrared spectroscopy and transcriptional studies. In vitro studies: Dose and time dependent cytotoxicity of agents were assessed against A431 cell line using MTT assay, LDH assay and acridine orange/ethidium bromide dual staining. Significant percentage decrease(s) in mean tumor volume and tumor burden were observed in nisin+ 5-FU combination treated groups as compared to alone treated groups. Histoarchitecture of treated skins demonstrated restoration towards normal skin tissue (being highest in the combination group). Modulation of apoptotic, angiogenic and proliferative genes were observed in treated groups. IC50 of combination was found to be 2 μg/ml as compared to nisin alone (32μg/ml) and 5-FU alone (16μg/ml) with combination index of 0.188. Dual staining showed that rate of induction of apoptosis was higher in the combination group as compared to single agents. Nisin and 5-FU in combination were found to be synergistic both in vivo and in vitro.


An example of 'naturally occurring' anti-Jsa (K6) in a Japanese female.
Anti-Jsa was found in a Japanese female who had never received a transfusion and had borne 3 Jsa-negative children. The antibody agglutinated Jsa cells suspended in saline and treated with proteolytic enzymes as well as reacting with those cells by the indirect antiglobulin test. This example is the first reported 'naturally occurring' anti-Jsa.


Combining automatic table classification and relationship extraction in extracting anticancer drug-side effect pairs from full-text articles.
Anticancer drug-associated side effect knowledge often exists in multiple heterogeneous and complementary data sources. A comprehensive anticancer drug-side effect (drug-SE) relationship knowledge base is important for computation-based drug target discovery, drug toxicity predication and drug repositioning. In this study, we present a two-step approach by combining table classification and relationship extraction to extract drug-SE pairs from a large number of high-profile oncological full-text articles. The data consists of 31,255 tables downloaded from the Journal of Oncology (JCO). We first trained a statistical classifier to classify tables into SE-related and -unrelated categories. We then extracted drug-SE pairs from SE-related tables. We compared drug side effect knowledge extracted from JCO tables to that derived from FDA drug labels. Finally, we systematically analyzed relationships between anti-cancer drug-associated side effects and drug-associated gene targets, metabolism genes, and disease indications. The statistical table classifier is effective in classifying tables into SE-related and -unrelated (precision: 0.711; recall: 0.941; F1: 0.810). We extracted a total of 26,918 drug-SE pairs from SE-related tables with a precision of 0.605, a recall of 0.460, and a F1 of 0.520. Drug-SE pairs extracted from JCO tables is largely complementary to those derived from FDA drug labels; as many as 84.7% of the pairs extracted from JCO tables have not been included a side effect database constructed from FDA drug labels. Side effects associated with anticancer drugs positively correlate with drug target genes, drug metabolism genes, and disease indications.


Unraveling the Heterogeneity of Hidradenitis Suppurativa with Phenotype Schema.
One of the major challenges faced with hidradenitis suppurativa (HS) is the variability in manifestations and treatment responses. Cazzaniga et al. (2020) conducted a cross-sectional study and latent class analysis to explain disease heterogeneity and formulate HS phenotypes. HS phenotypes might be useful for disease or treatment outcomes. Future studies should assess rater reliability and predictive validity for outcomes such as treatment response or disease progression.


Plasma treatment causes structural modifications in lysozyme, and increases cytotoxicity towards cancer cells.
Bacterial and mammalian proteins, such as lysozyme, are gaining increasing interest as anticancer drugs. This study aims to modify the lysozyme structure using cold atmospheric plasma to boost its cancer cell killing effect. We investigated the structure at acidic and neutral pH using various experimental techniques (circular dichroism, fluorescence, and mass spectrometry) and molecular dynamics simulations. The controlled structural modification of lysozyme at neutral pH enhances its activity, while the activity was lost at acidic pH at the same treatment conditions. Indeed, a larger number of amino acids were oxidized at acidic pH after plasma treatment, which results in a greater distortion of the lysozyme structure, whereas only limited structural changes were observed in lysozyme after plasma treatment at neutral pH. We found that the plasma-treated lysozyme significantly induced apoptosis to the cancer cells. Our results reveal that plasma-treated lysozyme could have potential as a new cancer cell killing drug.


Leachables from plastic materials in contact with drugs. State of the art and review of current analytical approaches.
The plastics used in drug packaging systems and medical devices are composed of homologous polymers and generally contain additives such as antioxidants, plasticizers and others, to improve their physicochemical properties. However, these additives have potential drawbacks due to possible migration or leaching towards the drug product. Leaching can cause a change in the chemical composition of the drug which, in turn, could modify its therapeutic action and, in some cases, its organoleptic properties. Leachables may also be considered a health hazard due to their inherent toxicological properties. The analytical characterization (detection, identification, typification/qualification and quantification) of leachable substances is mandatory and this information must be included in the application dossier for the drug before it can receive regulatory approval. The main aim of this paper is to collect and contextualise the reported analytical approaches for characterising and/or controlling organic leachables from plastic materials in contact with drugs. We also describe the state of the art of leachables in conjunction with a valuable, broad-based compilation of directives and guidelines. We end by presenting an updated collection of leachables both gas and liquid chromatography studies as separation techniques over the last eight years. We decided to focus our review exclusively on organic leachables as there is already a wide body of research on inorganic impurities.


The Effect of immunonutrition in patients undergoing pancreaticoduodenectomy: a systematic review and meta-analysis.
Pancreaticoduodenectomy (PD) is a complex and traumatic abdominal surgery with a high risk of postoperative complications. Nutritional support, including immunonutrition (IMN) with added glutamine, arginine, and ω-3 polyunsaturated fatty acids, can improve patients' prognosis by regulating postoperative inflammatory response. However, the effects of IMN on PD patients' outcomes require further investigation. PMC, EMbase, web of science databases were used to search literatures related to IMN and PD. Data such as length of hospital stay, infectious complications, non-infectious complications, postoperative pancreatic fistula (POPF), delayed gastric emptying (DGE), mortality, systemic inflammatory response syndrome (SIRS) duration, IL-6, and C-reactive protein (CRP) were extracted, and meta-analyses were performed on these data to study their pooled results, heterogeneity, and publication bias. This meta-analysis involved 10 studies and a total of 572 patients. The results showed that the use of IMN significantly reduced the length of hospital stay for PD patients (MD = -2.31; 95% CI = -4.43, -0.18; P = 0.03) with low heterogeneity. Additionally, the incidence of infectious complications was significantly reduced (MD = 0.42; 95% CI = 0.18, 1.00, P = 0.05), with low heterogeneity after excluding one study. However, there was no significant impact on non-infectious complications, the incidence of POPF and DGE, mortality rates, duration of SIRS, levels of IL-6 and CRP. The use of IMN has been shown to significantly shorten hospital stays and decrease the frequency of infectious complications in PD patients. Early implementation of IMN is recommended for those undergoing PD. However, further research is needed to fully assess the impact of IMN on PD patients through larger and higher-quality studies.


NCOA5 is a master regulator of amino acid-induced mTOR activation and β-casein synthesis in bovine mammary epithelial cells.
The nuclear receptor co-activator 5 (NCOA5) is known as a co-activator or co-repressor that influences gene expression and cellular physiology, but its roles and detailed molecular mechanism is still largely unknown. In this study, we explored the role and molecular mechanism of NCOA5 in amino acid-induced activation of the mechanistic target of rapamycin (mTOR) and milk protein synthesis in bovine mammary epithelial cells (BMECs). Methionine (Met) and leucine (Leu) significantly up-regulated the expression of NCOA5. NCOA5 overexpression increased mTOR phosphorylation and β-casein synthesis, whereas its knockdown exhibited the opposite effects. Furthermore, inhibition of phosphatidylinositol 3-kinase (PI3K) completely abolished the stimulatory effects of Met and Leu on NCOA5 expression. ChIP-qPCR analysis detected that NCOA5 bound to the mTOR promoter, and this interaction was enhanced by the stimulation of Met and Leu. These above data reveal that NCOA5 is a key regulator of amino acid-induced PI3K-mediated mTOR activation and β-casein synthesis in BMECs.


Mitochondrial nanomedicine: Subcellular organelle-specific delivery of molecular medicines.
As mitochondria network together to act as the master sensors and effectors of apoptosis, ATP production, reactive oxygen species management, mitophagy/autophagy, and homeostasis; this organelle is an ideal target for pharmaceutical manipulation. Mitochondrial dysfunction contributes to many diseases, for example, β-amyloid has been shown to interfere with mitochondrial protein import and induce apoptosis in Alzheimer's Disease while some forms of Parkinson's Disease are associated with dysfunctional mitochondrial PINK1 and Parkin proteins. Mitochondrial medicine has applications in the treatment of an array of pathologies from cancer to cardiovascular disease. A challenge of mitochondrial medicine is directing therapies to a subcellular target. Nanotechnology based approaches combined with mitochondrial targeting strategies can greatly improve the clinical translation and effectiveness of mitochondrial medicine. This review discusses mitochondrial drug delivery approaches and applications of mitochondrial nanomedicines. Nanomedicine approaches have the potential to drive the success of mitochondrial therapies into the clinic.


Efficient joint model learning, segmentation and model updating for visual tracking.
The Tracking-by-segmentation framework is widely used in visual tracking to handle severe appearance change such as deformation and occlusion. Tracking-by-segmentation methods first segment the target object from the background, then use the segmentation result to estimate the target state. In existing methods, target segmentation is formulated as a superpixel labeling problem constrained by a target likelihood constraint, a spatial smoothness constraint and a temporal consistency constraint. The target likelihood is calculated by a discriminative part model trained independently from the superpixel labeling framework and updated online using historical tracking results as pseudo-labels. Due to the lack of spatial and temporal constraints and inaccurate pseudo-labels, the discriminative model is unreliable and may lead to tracking failure. This paper addresses the aforementioned problems by integrating the objective function of model training into the target segmentation optimization framework. Thus, during the optimization process, the discriminative model can be constrained by spatial and temporal constraints and provides more accurate target likelihoods for part labeling, and the results produce more reliable pseudo-labels for model learning. Moreover, we also propose a supervision switch mechanism to detect erroneous pseudo-labels caused by a severe change in data distribution and switch the classifier to a semi-supervised setting in such a case. Evaluation results on OTB2013, OTB2015 and TC-128 benchmarks demonstrate the effectiveness of the proposed tracking algorithm.


Molecular screening for somatic mutations in corticotropic adenomas of dogs with pituitary-dependent hyperadrenocorticism.
Pituitary tumorigenesis is now generally regarded as a multistep process of genomic damage leading to uncoupling of interdependent systems that control cell proliferation and differentiation. The alterations include mutations in genes encoding for proteins involved in signal transduction pathways, such as G-proteins and the p21 protein encoded for by the ras genes. Apart from their excessive secretion of ACTH, corticotropic adenomas are characterized by decreased sensitivity to inhibition by glucocorticoids. Therefore, mutations in the glucocorticoid receptor leading to decreased sensitivity to glucocorticoids may contribute to corticotropic tumor formation. In this study, 16 corticotropic adenomas of dogs with pituitary-dependent hyperadrenocorticism were screened for mutations in the Gs alpha, H-, K-, N-ras genes and the coding region of the DNA-binding domain of the glucocorticoid receptor. The cDNA fragment of the Gs alpha gene encompassed codons 159-240. The K-, and N-ras fragments spanned codons 1-71. The H-ras gene was only screened for mutations in codons 12/13 by direct sequencing of the PCR product. The cDNA fragment of the DNA-binding domain of the glucocorticoid receptor encompassed codons 410-500. The Gs alpha, K-ras, N-ras genes and the DNA-binding domain of the glucocorticoid receptor were screened by single-strand conformation polymorphism analysis. No mutations were found in the Gs alpha gene, the ras genes and the DNA-binding domain of the glucocorticoid receptor. It is concluded that mutations in the Gs alpha gene (codons 159-240), the K- and N-ras genes (codons 1-71), the H-ras gene (codons 12/13) and mutations in the DNA-binding domain of the glucocorticoid receptor do not play a role in the tumorigenesis of canine corticotropic adenomas.


Angiopoietin-like 4 promotes glucose metabolism by regulating glucose transporter expression in colorectal cancer.
Angiopoietin-like 4 (ANGPTL4) was recently shown to be associated with cancer progression but little is known about its contribution to cancer metabolism. The purpose of this study was to elucidate the role of ANGPTL4 in glucose metabolism in colorectal cancer (CRC). Immunohistochemical staining of CRC specimens classified 84 patients into two groups according to ANGPTL4 expression. Clinicopathological characteristics, gene mutation status obtained by next-generation sequencing, and fluorodeoxyglucose (FDG) uptake measured by positron emission tomography/computed tomography (PET/CT) were compared between the two groups. Furthermore, the impact of ANGPTL4 expression on cancer metabolism was investigated by a subcutaneous xenograft mouse model using the ANGPTL4 knockout CRC cell line, and glucose transporter (GLUT) expression was evaluated. There were significantly more cases of T3/4 tumours (94.3% vs. 57.1%, P < 0.001) and perineural invasion (42.9% vs. 22.4%, P = 0.046) in the ANGPTL4-high group than in the low group. Genetic exploration revealed a higher frequency of KRAS mutation (54.3% vs. 22.4%, P = 0.003) in the ANGPTL4-high tumours. All the FDG uptake parameters were significantly higher in ANGPTL4-high tumours. In vivo analysis showed a significant reduction in tumour size due to ANGPTL4 knockout with lower expression of GLUT1 and GLUT3, and suppression of AKT phosphorylation. ANGPTL4 regulates the expression of GLUTs by activating the PI3K-AKT pathway and thereby promoting glucose metabolism in CRC. These findings establish a new functional role of ANGPTL4 in cancer progression and lay the foundation for developing a novel therapeutic target.


[Relationship between individual radiographic findings and disability in rheumatoid arthritis].
To evaluate if the duration of disease influences the link between different radiographic specific features and disability in rheumatoid arthritis (RA) and the influence of disease duration on this relationship. Conventional X- rays of both hands of 96 patients with RA were evaluated independently by 2 readers using Kayes' modification of the Sharp method. Disability was evaluated with the Spanish version of the HAQ questionnaire. The mean HAQ was 1, 39 ± 0, 79. The mean total radiographic score was 0.8 (18% of the maximum possible score). Total and joint space narrowing scores only displayed a statistically significant correlation (r=0.33, r=.37, respectively, P<.05) with disability in the late RA group (>7 years). Erosion and malalignment scores were not correlated with HAQ. There was a statistically significant correlation between the eating, dressing and reach HAQ-categories and the total radiographic score in the late RA group (r=0.48, P<.001, r=0.42, P<.01, r=0.3, P<.05, respectively). This work suggests that HAQ disability and radiographic damage are only related in cases with late RA. In this group, the subtotal radiographic score most related with disability is the joint space narrowing score.


Prolactin binding sites in the brain and kidneys of the toad, Bufo arenarum Hensel.
(125I)-ovine prolactin (oPRL) binding was found in several brain areas of the toad, Bufo arenarum Hensel. The olfactory bulb, cerebral hemispheres, and both dorsal and ventral mesencephalic regions showed saturable, high affinity, (125I)-oPRL binding, ranging between 5.6 to 29.9 fmol/mg protein, while the association constant (Ka) by Scatchard analysis was between 4.0 to 8.7 x 10(9) M-1. This binding was compared with the Scatchard plot of the kidney, which has been already described by other groups, and gave 41.7 fmol/mg protein and Ka 2.5 x 10(9) M-1. Liver showed no binding and in the cerebral hemispheres (125I)-oPRL was not displaced by non-lactogenic hormones, indicating that binding was hormone and tissue specific.


A large pericardial cyst mimicking a unilateral pleural effusion: A case report.
Pericardial cysts are a rare benign disorder with a variable clinical presentation depending on their size and location. The diagnosis of pericardial cysts is usually based on imaging examinations. The definitive treatment is surgical resection. A 36-year-old woman presented with progressive left-sided chest pain and exertional dyspnea, with symptoms resembling pleural effusion. The patient was diagnosed with a pericardial cyst based on imaging and video-assisted thoracoscopic surgery (VATS). VATS was performed. The patient's symptoms improved after successful removal of the pericardial cyst. Follow-up chest computed tomography exhibited no evidence of recurrence. Clinicians should include pericardial cysts in the differential diagnosis of pseudopleural effusion. VATS is a feasible and safe method to treat symptomatic and large pericardial cysts.


Objective signs of inferior alveolar nerve anesthesia in exodontia.
Objective determination of the adequacy of inferior alveolar nerve anesthesia is difficult. A dependable objective test based upon clinical observation of hundreds of patients during extraction of mandibular teeth, is presented. Following routine inferior alveolar nerve block anesthesia, time is allowed for subjective signs of "tingling" of the lower lip. Fully aware that the patient may respond to pressure in the absence of pain, the tooth to be extracted is tested. Gradual pressure of increasing intensity is applied with extracting forceps for 3 seconds, then suddenly released, providing a sudden stimulus. If the patient immediately moves or "jumps," this response indicates inadequate anesthesia. If the patient does not respond to the sudden release of pressure, that tooth can be removed painlessly even though the patient may still respond to pressure.


Stimuli-responsive nanocarriers for bacterial biofilm treatment.
Bacterial biofilm infections have been threatening the human's life and health globally for a long time because they typically cause chronic and persistent infections. Traditional antibiotic therapies can hardly eradicate biofilms in many cases, as biofilms always form a robust fortress for pathogens inside, inhibiting the penetration of drugs. To address the issues, many novel drug carriers emerged as promising strategies for biofilm treatment. Among them, stimuli-responsive nanocarriers have attracted much attentions for their intriguing physicochemical properties, such as tunable size, shape and surface chemistry, especially smart drug release characteristic. Based on the microenvironmental difference between biofilm infection sites and normal tissue, many stimuli, such as bacterial products accumulating in biofilms (enzymes, glutathione, etc.), lower pH and higher H2O2 levels, have been employed and proved in favor of "on-demand" drug release for biofilm elimination. Additionally, external stimuli including light, heat, microwave and magnetic fields are also able to control the drug releasing behavior artificially. In this review, we summarized recent advances in stimuli-responsive nanocarriers for combating biofilm infections, and mainly, focusing on the different stimuli that trigger the drug release. , , 。 , , 。 , , 。 , -, , , , 。 , , (, ), pHH2O2, ""。 , , , , 。 , , 。.


The construction of a novel xenograft bovine bone scaffold, (DSS)6-liposome/CKIP-1 siRNA/calcine bone and its osteogenesis evaluation on skull defect in rats.
Xenograft bone scaffolds have advantages such as mechanical strength, sufficient source and safety. Combined with siRNA properly targeting CKIP-1, a negative regulator of osteogenesis, may contribute to the repair result of calcine bone alone. Herein, we constructed a novel xenograft bovine bone scaffold namely (DSS)6-liposome/CKIP-1 siRNA/calcine bone, the characteristics of which were investigated by confirming the effect of (DSS)6-liposome, observing the appearance and testing mechanical strength of calcine bone, and observing the combined result of CKIP-1 siRNA by FAM immunofluorescence. In addition, cytotoxicity by CCK-8 and LDH activity of L929 cells and MC3T3-E1 osteoblasts cultured with the scaffold were tested in vitro, primary osteoblasts proliferation, the mRNA expressions of CKIP-1, ALP, COL1-α and OCN, the protein expressions of CKIP-1, BMP-2, COL-1 and Runx2 and calcium nodules were also determined by CCK-8, RT-qPCR, western-blot and Alizarin Red staining in vitro. Then, we successively established the skull defect model for evaluating the repair result of the novel scaffold by HE staining of 2, 4, 8 and 12 weeks, immumohistochemical stainings of 2, 4, 8 and 12 weeks such as ALP, COL-1α and OCN, Mirco-CT scanning of 4 and 12 weeks and the relative parameters and so on in vivo. It indicated that (DSS)6-liposome/CKIP-1 siRNA/calcine bone could successfully knock down the CKIP-1 mRNA and protein expressions, promote osteoblasts proliferation with the little cytotoxicity in vitro, increase the protein expressions of BMP-2, COL-1 and Runx2 in vitro, increase mRNA expressions of ALP, COL-1α and OCN in vitro and in vivo, and have a better bone defect repair effect with few side effects in rats after 12 weeks. Our research indicates (DSS)6-liposome/CKIP-1 siRNA/calcine bone could repair skull defects well in rats, and it may lay the foundation of applicating the novel xenograft bone scaffold in the clinical. These findings provide evidence that (DSS)6- liposome/CKIP-1 siRNA/calcine bone could be used as a novel xenograft bone scaffold for osteogenesis with the good safety.


Results of oophorectomy by menstrual and estrogen receptor states in patients with metastatic breast cancer.
The records of 204 women with metastatic breast carcinoma treated by oophorectomy were analyzed. Premenopausal women had a response rate of 50 percent. Forty-one percent of postmenopausal women responded. Those who responded had an average duration of response of 22 months and a length of survival twice that of the nonresponders. There was a better than 60 percent correlation between response to oophorectomy and response to further endocrine ablation. Response to endocrine manipulation is more a function of the hormonal sensitivity of the carcinoma than of menopausal status.


Complete genome sequences of three newly discovered cacao mild mosaic virus isolates from Theobroma cacao L. in Brazil and Puerto Rico and evidence for recombination.
To analyze the DNA virome associated with cacao (Theobroma cacao L.) trees showing virus-like symptoms in Brazil (BR) and Puerto Rico (PR) during 2018-2019, total DNA was isolated from symptomatic leaves and subjected to high-throughput Illumina sequencing. The assembled complete badnaviral genome sequences were verified by PCR amplification, cloning, and DNA sequencing. Based on pairwise distances and phylogenetic analysis, three badnaviral genomes were identified, and these viruses were found to be isolates of the previously described cacao mild mosaic virus (CaMMV). The three genomes were 7,520, 7,524, and 7,514 bp in size for the isolates CaMMV-BR321, CaMMV-BR322, and CaMMV-PR3, respectively. Each genome contained four predicted open reading frames: ORFs 1-3 and ORFY. The CaMMV-PR3 isolate was identified as a probable recombinant, with a CaMMV-BR-like virus as the major parent.


Purification and characterization of recombinant Cel7A from maize seed.
The corn grain biofactory was used to produce Cel7A, an exo-cellulase (cellobiohydrolase I) from Hypocrea jecorina. The enzymatic activity on small molecule substrates was equivalent to its fungal counterpart. The corn grain-derived enzyme is glycosylated and 6 kDa smaller than the native fungal protein, likely due to more sugars added in the glycosylation of the fungal enzyme. Our data suggest that corn seed-derived cellobiohydrolase (CBH) I performs as well as or better than its fungal counterpart in releasing sugars from complex substrates such as pre-treated corn stover or wood. This recombinant protein product can enter and expand current reagent enzyme markets as well as create new markets in textile or pulp processing. The purified protein is now available commercially.


Development and Application of a Quantitative PCR Detection Method to Quantify Venturia oleaginea in Asymptomatic Olive (Olea europaea) Leaves.
Olive leaf spot (OLS), caused by Venturia oleaginea, is one of the most common and serious diseases of olive trees in the Mediterranean region. Understanding the pathogen life cycle is important for the development of effective control strategies. Current knowledge is incomplete owing to a lack of effective detection methods. It is extremely difficult to culture V. oleaginea in vitro, so primers were designed to amplify and sequence the internal transcribed spacer ITS1-5.8S-ITS2 region of the fungus directly from infected olive leaves. Sanger sequencing indicated a unique ITS region present in the European strains screened, confirming the appropriateness of the target region for developing a quantitative PCR (qPCR) assay. Furthermore, high-throughput sequencing of the same region excluded the presence of other Venturia species in the olive phyllosphere. The qPCR assay proved very specific and sensitive, enabling the detection of approximately 26 copies of target DNA. The analysis of symptomless leaves during early stages of the epidemic from the end of winter through spring revealed a similar quantity of pathogen DNA regardless of the leaf growth stage. In contrast, the pathogen titer changed significantly during the season. Data indicated that leaf infections start earlier than expected over the season and very young leaves are as susceptible as adult leaves. These findings have important practical implications and suggest the need for improved scheduling of fungicide treatments. The qPCR assay represents a valuable tool providing quantitative results and enables detection of V. oleaginea in all olive organs, including those in which OLS cannot be studied using previously available methods.


First-time-in-human dose selection: allometric thoughts and perspectives.
Some of the many factors that influence dose selection in first-time-in-human studies are examined. These include animal toxicology, toxicokinetics, allometric scaling, pharmacokinetics, body surface area correlations, and integration of preclinical pharmacologic and toxicologic data. Appropriate preclinical evaluation and analysis may reduce the frequency and severity of unexpected toxic events arising during single-dose, phase I testing. However, significant intrinsic uncertainties in this process presently exist and will continue to exist well into the foreseeable future. With our present state of knowledge, we cannot provide a realistic and reasonable algorithm for ascertaining first-time-in-human doses: any decision tree would be too unwieldy. There are several rules of thumb that do have a place in the evaluation and decision-making process, however.


Compliance in the elderly.
Noncompliance is not a unique phenomenon associated with aging. Many of the same problems concerning taking medications most appropriately apply to 40 year olds as well as to 80 year olds. What is unique about the elderly is their greater sensitivity to medications, their greater propensity for the development of adverse effects, and the greater complexity of their regimens as they develop chronic illnesses through the course of their lives. In addition, decreasing acuity of special senses, problems with memory, and the interposition of other caregivers are all unique phenomena that have to be dealt with through expansion of the standard methods but only rarely through specific methods. Utilization of modern technology such as video cassette recorders, large print reproductions, and improved communication modalities for people with hearing and sight disabilities may overcome some of the problems leading to decreased comprehension and communication ability in elderly patients. Similarly, for those with financial barriers to appropriate utilization of medications, programs such as the Elderly Pharmaceutical Insurance Coverage (EPIC) in New York State may be helpful. Medicaid and AARP programs for obtaining medications at lower prices can help financially strapped patients. The necessity for physicians to learn about cultural attitudes toward family and governmentally supplied caregivers may also facilitate optimal medication utilization in our polycultural society. Medications can serve the elderly to great benefit. Misapplication and misuse of medications may result in undesirable outcomes, ranging from lack of maximum efficacy, annoying nuisance side effects, diminishing quality-of-life, and even life-threatening adverse outcomes.(ABSTRACT TRUNCATED AT 250 WORDS)


Trends and Contributing Factors for Prescribing Antipsychotics in Newly Diagnosed Parkinson's Disease Patients: A Population-Based Study.
Psychosis is very common in later stages of Parkinson's disease (PD) patients, but its prevalence in newly diagnosed patients is not rare. The use of antipsychotics in PD patients is complex given their association with worsening Parkinsonian motor symptoms and increased mortality rate. This study aims to examine factors that affect the use of antipsychotics in newly diagnosed PD patients and to identify changes in prescribing over time. The Secure Anonymized Information Linkage (SAIL) databank was used to identify a cohort of newly diagnosed PD patients aged 40 years and older in Wales. The cohort was longitudinally examined over 17 years to determine the incidence of new antipsychotic use. Logistic regression models were used to analyze the data and were adjusted for several potential confounding variables. A total of 9142 PD patients were identified after applying inclusion and exclusion criteria, of whom 58.6% were male. During the first year of PD diagnosis, 12% of the patients developed psychosis and were prescribed antipsychotics. Quetiapine was the most commonly prescribed (49%), followed by risperidone (10.7%). The use of antipsychotics in newly diagnosed PD patients was significantly lower in the years 2009-2016 compared to 2000-2008 (OR 0.37, 95% CI 0.32-0.43). Other significant prescribing factors included patient's age and history of dementia. A dramatic decline in antipsychotic use was identified across years, showing adherence to warnings against use of antipsychotics for PD patients. Given that psychosis is prevalent in PD patients, the continuous assessment of the safety risks of antipsychotics is a matter of priority.


Adaptive R&D contract for urgently needed drugs: Lessons from COVID-19 vaccine development.
This paper analyzes an incentive contract for new vaccine research and development (R&D) under pandemic situations such as COVID-19, considering the R&D contract's adaptability to the pandemic. We study how the public sector (government) designs the adaptive R&D contract and offers it to pharmaceutical enterprises. An agency-theoretic model is employed to explore the contract whose terms are an upfront grant as a fixed fee and a sales tax credit as an incentive tool, examining how the values of related parameters affect contract term determinations. We found that the adaptability factor derived from urgent policies such as emergency use authorization (EUA) as well as tax credits, can be utilized as practical incentive tools that lead vaccine developers to increase their effort levels for R&D success. We also found that public-private state-emergency contracts may not follow the conventional wisdom. Counterintuitively, dependency on tax credits (incentive part) decrease as the client's degree of risk averseness increases in the emergency contract.


The combined recombinant cathepsin L1H and cathepsin B3 vaccine against Fasciola gigantica infection.
Protections against Fasciola gigantica infection in mice immunized with the individual and combined cathepsin L1H and cathepsin B3 vaccines were assessed. The vaccines comprised recombinant (r) pro-proteins of cathepsin L1H and B3 (rproFgCatL1H and rproFgCatB3) and combined proteins which were expressed in Pichia pastoris. The experimental trials were performed in ICR mice (n = 10 per group) by subcutaneous injection with 50 μg of the recombinant proteins combined with Alum or Freund's adjuvants. At two weeks after the third immunization, mice were infected with 15 F. gigantica metacercariae per mouse by oral route. The percents of protection of rproFgCatL1H, rproFgCatB3 and combined vaccines against F. gigantica were approximately 58.8 to 75.0% when compared with adjuvant-infected control. These protective effects were similar among groups receiving vaccines with Alum or Freund's adjuvants. By determining the levels of IgG1 and IgG2a in the immune sera, which are indicative of Th1 and Th2 immune responses, it was found that both Th1 and Th2 humoral immune responses were significantly increased in vaccinated groups compared with the control groups, with higher levels of IgG1 (Th2) than IgG2a (Th1). Mice in vaccinated groups showed reduction in liver pathological lesions when compared with control groups. This study indicates that the combined rproFgCatB3 and rproFgCatL1H vaccine had a high protective potential than a single a vaccine, with Alum and Freund's adjuvants showing similar level of protection. These results can serve as guidelines for the testing of this F. gigantica vaccine in larger economic animals.


The development and evaluation of an oncological palliative care deprescribing guideline: the 'OncPal deprescribing guideline'.
Current data suggests that potentially inappropriate medicines (PIMs) are common in palliative cancer patients; however, there is a lack of criteria to assist clinicians in identifying PIMs in these patients. The aims of this study were to design and validate a deprescribing guideline for palliative cancer patients and to undertake a descriptive analysis of the identified PIMs. This prospective, non-interventional cohort study consisted of four major stages: developing an 'OncPal Deprescribing Guideline' from current evidence, the prospective recruitment of consecutive palliative cancer inpatients with an estimated <6-month prognosis, the assessment of all medications to identify PIMs using both a panel of medical experts without access to the guideline as well as a Clinical Pharmacist independently using the OncPal Deprescribing Guideline and the evaluation of the guideline by testing concordance. Descriptive data on the incidence of PIMs identified were also assessed. A total of 61 patients were recruited. The OncPal Deprescribing Guideline matched 94% of 617 medicines to the expert panel with a Kappa value of 0.83 [95% CI (0.76, 0.89)] demonstrating an 'outstanding' concordance. Forty-three (70%) patients were taking at least one PIM, with 21.4% of the total medicines assessed identified as PIMs. The medication-associated cost per patient/month was AUD$26.71. A guideline to assist in the de-escalation of inappropriate medications in palliative cancer patients was developed from current literature. The OncPal Deprescribing Guideline was successfully validated, demonstrating statistically significant concordance with an expert panel. We found that the incidence of PIMs was high in our patient group, demonstrating the potential benefits for the OncPal Deprescribing Guideline in clinical practice.


Localization of the active site of human tumour necrosis factor (hTNF) by mutational analysis.
In order to define the active site(s) of human tumour necrosis factor (hTNF), we mutagenized its gene at random and directly screened the resulting population for loss of cytotoxic activity on L929 cells. Four biologically inactive mutant proteins (Arg32----Trp, Leu36----Phe, Ser86----Phe and Ala84----Val) behaved similar to the wild-type in various physico-chemical assays. The residues were positioned on a 3D structural model and were found to cluster together at the base of the molecule at each side of the groove that separates two monomers in the trimeric structure. A very conservative mutation at one of these sites (Ala84----Val) almost completely abolished cytotoxic activity. Amino acid alterations in three other residues in close proximity to this receptor binding site were introduced: replacements at positions 29 and 146 clearly reduced cytotoxicity only when non-conservative alterations were introduced (Leu29----Ser and Glu146----Lys), suggesting an indirect influence on the active site. However, a conservative mutation at position 91 (Val----Ala) caused a significant drop (500-fold) in bioactivity which suggests that Val91 may also play a direct role in receptor recognition. Our results favor a model in which each TNF molecule has three receptor-interaction sites (between the three subunits), thus allowing signal transmission by receptor clustering.


Generation of a transgene-free induced pluripotent stem cell line (SMBCi011-A) from a patient with Prader-Willi syndrome.
Prader-Willi syndrome (PWS) is a neurodevelopmental disorder caused by deficiency of paternal gene expression in the 15q11.2-q13 chromosome imprinted region. Hyperphagia and dysgnosia are typical clinical features in the early-childhood of patient. We generated an induced pluripotent stem cell (iPSC) line SMBCi011-A from a 6 years old male PWS patient, the line expressed pluripotent signs and had ability to differentiate into three germ layers in vivo.


Predictive value of rTMS in the identification of responders to epidural motor cortex stimulation therapy for pain.
This study was designed to assess the value of repetitive transcranial magnetic stimulation (rTMS) to predict the efficacy of epidural motor cortex stimulation (EMCS) to treat neuropathic pain. We have included 59 patients treated by EMCS for more than 1 year and in whom active and sham 10Hz-rTMS sessions were performed as preoperative tests, targeted over the cortical representation of the painful area. Analgesic effects were rated on a visual analogue scale. The real rTMS efficacy was determined by subtracting the effect of the sham stimulation on pain scores from that of the active stimulation (active-sham calculation). Pain scores were significantly reduced by active rTMS and EMCS, but not by sham rTMS. Twenty-six of the 33 patients (79%) who responded to active rTMS and all the 21 patients (100%) who responded for active-sham calculation also responded to EMCS. The response observed in active-sham calculation had a positive predictive value of 1.0, but a negative predictive value of .6 regarding EMCS outcome. The analgesic effect of rTMS or EMCS was not influenced by the side, origin, or duration of pain or by the presence of motor or sensory deficit in the painful area. Poorer results were observed in case of lower limb pain for rTMS and in older patients for EMCS. This study confirms that neuropathic pain can be significantly relieved by motor cortex rTMS or EMCS. A positive outcome of EMCS can be predicted by a real response to rTMS, but not on clinical grounds. Single sessions of sham-controlled preoperative rTMS tests can be used to confirm the indication of EMCS therapy but have no value to exclude patients from this therapy. New rTMS protocols remain to be assessed to improve the usefulness of preoperative rTMS in EMCS practice.


A new species of Umanella Gauld (Hymenoptera: Ichneumonidae: Pimplinae) from Colombia.
A new species of Umanella Gauld 1991, U. tricoloripes Herrera-Florez, sp. n., is described from two males collected in Colombia. Umanella tricoloripes sp. nov. differs substantially from the other species of the genus in color pattern.