Hypertonic saline cough provocation test with salbutamol pre-treatment: evidence for sensorineural dysfunction in asthma.
Cough is one of the most common symptoms of asthma. However, studies using capsaicin, citric acid, or tartaric acid to document cough threshold have repeatedly failed to show statistically significant differences between asthmatic and healthy subjects. The studies using hypertonic aerosols as the cough stimulant have suggested an enhanced sensitivity in asthmatic subjects but the induced bronchoconstriction has made the interpretation of the results difficult. To determine the cough sensitivity to hypertonicity in healthy subjects, patients with chronic cough, and patients with asthma in a setting where the induction of bronchoconstriction is prevented. Nineteen healthy subjects, 21 non-asthmatic patients with chronic cough, and 26 asthmatic patients with chronic cough underwent an incremental hypertonic saline challenge including a pre-treatment with 0.4 mg of salbutamol. Spirometry was performed before the challenge, after salbutamol, and after the challenge. The patients with cough also underwent skin testing, histamine challenge, exhaled nitric oxide measurement, ambulatory peak flow monitoring, kept cough diary, and filled in the Leicester Cough Questionnaire. Eighteen patients repeated the saline challenge. The challenge did not induce bronchoconstriction in any group. The osmolality to provoke 15 cumulative coughs was significantly smaller in the asthmatic patients than in the healthy subjects (P<0.001) and in the cough patients without asthma (P=0.04). According to multivariate analysis among all the 47 patients with cough, female sex (P<0.001) and large spontaneous peak flow variation in the ambulatory recording (P=0.001) were associated with high sensitivity to saline. The saline challenge response was well repeatable (intraclass correlation coefficient 0.90). The findings of the present study are not affected by induced bronchoconstriction. Asthma or, more specifically, spontaneous, reversible airway obstruction is associated with an enhanced sensitivity to the cough-provoking effect of hypertonic saline. This suggests a pathological function of the sensorineural apparatus in this disorder.


Clinical predictors of mechanical ventilation in Fisher/Guillain-Barré overlap syndrome.
Some patients with Fisher syndrome (FS) developed subsequent descending tetraparesis (Fisher/Guillain-Barré overlapping syndrome: FS/GBS). The assumption is that such descending progression may frequently lead to respiratory failure. To investigate whether patients with FS/GBS more often require artificial ventilation than those with typical GBS and which clinical and serological findings are useful predictors. Medical records were reviewed of patients who had acute ophthalmoplegia, ataxia and areflexia, as well as subsequent tetraparesis with monophasic course. Forty-five patients fulfilled the FS/GBS criteria. Clinical and serological features were analysed, and clinical predictors of mechanical ventilation were investigated. FS/GBS patients more frequently required mechanical ventilation than did GBS patients (24% vs 10%, p = 0.04). The former also needed artificial ventilation earlier than the latter (p = 0.03), but none of the FS patients required it. As the initial symptom, ventilated FS/GBS patients more frequently showed titubation than non-ventilated patients (55% vs 18%, p = 0.04). During the course of the illness, descending tetraparesis was more common in 11 ventilated FS/GBS patients than in the other 34 non-ventilated patients (64% vs 21%, p = 0.02). The need for artificial ventilation was not associated with anti-GQ1b IgG antibodies, monospecific anti-GT1a IgG antibodies or IgG antibodies to various ganglioside complexes. FS/GBS patients significantly needed mechanical ventilation more often. Such patients showing titubation and descending tetraparesis need to be carefully monitored as the illness progresses because those clinical features are helpful predictors of respiratory failure.


Antihormonal treatment of paraphilic patients in German forensic psychiatric clinics.
The aim of this study was to investigate which antihormonal treatment strategies are used in German forensic psychiatric institutions. Forensic clinics were asked about the number of treated patients. Four hundred seventy-four patients were committed for sex offences; 12% received either CPA (n = 29) or LHRH- agonists (n = 29). Differences in efficacy were small. Several side effects confirm the importance of a protocol for minimizing medical complications.


Wave-Function Symmetry Control of Electron-Transfer Pathways within a Charge-Transfer Chromophore.
Despite a diverse manifold of excited states available, it is generally accepted that the photoinduced reactivity of charge-transfer chromophores involves only the lowest-energy excited state. Shining a visible-light laser pulse on an aqueous solution of the chromophore-quencher [Ru(tpy)(bpy)(μNC)OsIII(CN)5]- assembly (tpy = 2,2';6,2''-terpyridine and bpy = 2,2'-bipyridine), we prepared a mixture of two charge-transfer excited states with different wave-function symmetry. We were able to follow, in real time, how these states undergo separate electron-transfer reaction pathways. As a consequence, their lifetimes differ in 3 orders of magnitude. Implicit are energy barriers high enough to prevent internal conversion within early excited-state populations, shaping isolated electron-transfer channels in the excited-state potential energy surface. This is relevant not only for supramolecular donor/acceptor chemistry with restricted donor/acceptor relative orientations. These energy barriers provide a means to avoid chemical potential dissipation upon light absorption in any molecular energy conversion scheme, and our observations invite to explore wave-function symmetry-based strategies to engineer these barriers.


Gastrointestinal endoscopic surgery.
A working knowledge of normal endoscopic anatomy and the effects of positioning must be mastered to take advantage of endoscopic procedures involving the gastrointestinal system. Endoscopic procedures involving the gastrointestinal system offer advantages over conventional surgery in that they are less invasive and provide direct visualization. Laparoscopy can be used to evaluate horses with abdominal pain before and after surgery. It can be used to evaluate and biopsy abdominal masses and parenchymal organs such as the spleen, liver, and kidney. Endoscopic gastrointestinal surgical procedures such as colopexy and adhesionolysis are limited at this time but should continue to be developed as instrumentation and technology evolve.


Correction to: Retransplantation after post transplant lymphoproliferative disorder: overcoming the obstacles!
In the original publication of the article, while submitting the case report.


Bacterial chaperone protein, Skp, induces leukocyte chemotaxis via C5a receptor.
C5a receptor has been identified as a leukocyte chemotactic receptor to two intrinsic chemical mediators, C5a and the S19 ribosomal protein dimer, so far. We found an Escherichia coli protein that also induced the chemotactic responses of monocytes and polymorphonuclear leukocytes via the C5a receptor. We identified the E. coli-derived chemoattractant to be Skp by the molecular size and the N-terminal amino acid sequence. Skp is a periplasmic chaperone protein widely present in gram-negative bacterial species. Immunoabsorption experiments indicated that Skp was the major leukocyte chemotactic factor in the E. coli extract. Receptor-antagonizing experiments with analogue peptides of S19 ribosomal protein and of C5a suggested that Skp induced the receptor activation by the two-step binding mechanism as in the cases of the intrinsic mediators, sharing the ligand-binding site of the receptor among them at each binding step. The C5a receptor would play a role in the host defense directly recognizing the bacteria-derived protein, besides identifying the signals of the intrinsic chemical mediators.


Involvement of the p110 alpha isoform of PI3K in early development of mouse embryos.
Class I of phosphoinositide 3-kinases (PI3Ks) is characterized as a group of intracellular signal proteins possessing both protein and lipid kinase activities. Recent studies implicate class I of PI3Ks acts as indispensable mediators in early development of mouse embryos, but the molecular mechanisms are poorly defined. In this paper, mouse one-cell embryos were used to investigate a possible contribution of the catalytic subunit of PI3K, p110 alpha, to cell cycle progression. The expression level of p110 alpha was determined in four phases of one-cell embryos. Silencing of p110 alpha by microinjection of p110 alpha shRNA into one-cell embryos resulted in a G2/M arrest and prevented the activation of Akt and M-phase promoting factor (MPF). Further, microinjection of the synthesized mRNA coding for a constitutively active p110 alpha into one-cell embryos induced cell cleavage more effectively than microinjection of wild-type p110 alpha mRNA, whereas microinjection of mRNA of kinase-deficient p110 alpha delayed the first mitotic cleavage. Taken together, this study demonstrates that p110 alpha is significant for G2/M transition of mouse one-cell embryos and further emphasizes the importance of Akt in PI3K pathway.


Effect of clothing weight on body weight.
In clinical settings, it is common to measure weight of clothed patients and estimate a correction for the weight of clothing, but we can find no papers in the medical literature regarding the variability in clothing weight of adults with weather, season and gender. Fifty adults (35 women) were weighed four times during a 12-month period with and without clothing. Clothing weights were determined and regressed against minimum, maximum and average daily outdoor temperature. The average clothing weight (±s.d.) throughout the year was significantly greater in men than in women (1.2±0.3 vs 0.8±0.3 kg, P<0.0001). The average within-person minimum and the average within-person maximum clothing weights across the year were 0.9±0.2 and 1.5±0.4 kg for men, and 0.5±0.2 and 1.1±0.4 kg for women, respectively. The within-person s.d. in clothing weight was 0.3 kg for both men and women. Over the 55 °C range in the lowest to the highest outdoor temperatures, the regressions predicted a maximal change in clothing weight of only 0.4 kg in women and 0.6 kg in men. The clothing weight of men is significantly greater than that of women, but there is little variability throughout the year. Therefore, a clothing adjustment of approximately 0.8 kg for women and 1.2 kg for men is appropriate regardless of outdoor temperature.


[The effect of polyamine on the structure of the free amino acid pool in the blood plasma and in the liver of animals with chronic alcoholic intoxication].
The effect of polyamine on the total content of free amino acids and the structure of the amino acid fund in blood plasma and hepatic tissue was studied in rats with chronic alcoholic intoxication. Rapid utilization of exogenic introduced amino acids and a normalizing effect of the drug on the structure of the amino acid fund were demonstrated. The high concentration of aromatic amino acids in the drug may be among the causes of the increased content of this fraction of amino acids in the tissues studied after administration of the drug.


Replication patterns and cytopathology of cells infected with baculoviruses.
In vitro studies have contributed greatly to an understanding of viral cytopathology, molecular biology, and pathogenesis. A model of the role of baculoviruses in a host-parasite relationship is developing which reveals the virus as gaining control of many aspects of host cell biology including control of the cell replication machinery (apoptotic response, macromolecular synthesis), the cytoskeletal structure, the nuclear membrane and intranuclear architecture. Baculovirus replication is a collection of independent but inter-related processes which work within the framework of the host cell, with the in vivo goal of maximizing production of progeny virions. Further molecular dissection of baculovirus replication should yield insight into the processes and principles of viral and host regulatory systems, perhaps facilitating development of new generations of high efficiency sub-viral expression vector systems and the development of genetically improved strains of virus safe for field use in ecologically based pest management strategies.


Akt phosphorylation is required for heat acclimation-induced neuroprotection.
Long-term heat exposure, known as heat acclimation (HA; 30 days at 34 +/- 1 degrees C) is neuroprotective against traumatic brain injury. Acclimated mice were previously found to display improved functional recovery as well as an increase in the levels of the specific erythropoietin receptor. As the activation of this receptor is known to facilitate functional recovery on one hand and the phosphorylation and activation of Akt, an intracellular kinase which regulates anti-apoptotic pathways on the other, in this study we investigated whether HA affects Akt phosphorylation prior to and following injury and whether this step is required for development of HA-induced neuroprotection. Akt phosphorylation was blocked using Triciribine (TCN), a compound shown to block the phosphorylation process without affecting upstream effectors of this kinase, and several post-injury functional end-point measures were subsequently evaluated. Acclimation led to a post-injury increase in the levels of phosphorylated Akt, resulting in higher levels when compared with normothermic controls at 4 h post-injury (63.6 +/- 5.2% and 42.7 +/- 3.7%, respectively, p </= 0.05). This increase was diminished following TCN administration. Post-injury TCN treatment abolished the HA-induced functional benefits, including effects on motor and cognitive functions as well as the attenuation of edema formation. We therefore suggest that Akt phosphorylation is essential for HA-induced neuroprotection after traumatic brain injury.


Effect of co-solutes and process variables on crystallinity and the crystal form of freeze-dried myo-inositol.
The purpose of this study was to elucidate how co-solutes affect the crystallization of small solute molecules during freeze-drying and subsequent storage. Crystallization profiles of myo-inositol and its mixture with dextran 40k in frozen solutions and dried solids were assessed by thermal analysis (DSC), powder-X-ray diffraction, and simultaneous DSC and PXRD analysis. Higher mass ratios of dextran maintained myo-inositol in the non-crystalline mixture state, in frozen solutions, during freeze-drying process, and exposure of dried solids to higher temperatures. Co-lyophilization with a lower mass ratio of dextran resulted in solids containing a variety of myo-inositol crystal forms and crystallinity depending on the composition and thermal history of the process. Heating of some inositol-rich amorphous solids showed crystallization of myo-inositol in the metastable form and its transition to stable form before melting. Heat-treatment of inositol-rich frozen solutions resulted in high crystallinity stable-form inositol solids, leaving dextran in the amorphous state. Sufficient direct molecular interactions (e.g., hydrogen bonding) should explain the stability of dextran-rich amorphous solids. Optimizing solute composition and processes should be a potent way to control crystal form and crystallinity of components in freeze-dried formulations.


Radioactivity in mushrooms, mosses and soil samples of defined biotops in SW Bavaria-two years after "Chernobyl".
Mushrooms, the moos/grass layer and soil samples have been collected in autumn 1987 from two sites in SW-Bavaria in continuation of a former investigation (Elstner et al. 1987). There were still relatively high amounts of 137Cs and 134Cs in all samples. The ratio 137Cs/134Cs changed according to the different half-life times of the two radioisotopes, indicating in nearly all cases the Chernobyl accident as source. The distribution of the radioisotopes within the mushroom populations shows considerable variation, even within the same species and location. Besides 137Cs, 134Cs and 40K no other radioisotopes were detected.


Iron overload cardiomyopathy: better understanding of an increasing disorder.
The prevalence of iron overload cardiomyopathy (IOC) is increasing. The spectrum of symptoms of IOC is varied. Early in the disease process, patients may be asymptomatic, whereas severely overloaded patients can have terminal heart failure complaints that are refractory to treatment. It has been shown that early recognition and intervention may alter outcomes. Biochemical markers and tissue biopsy, which have traditionally been used to diagnose and guide therapy, are not sensitive enough to detect early cardiac iron deposition. Newer diagnostic modalities such as magnetic resonance imaging are noninvasive and can assess quantitative cardiac iron load. Phlebotomy and chelating drugs are suboptimal means of treating IOC; hence, the roles of gene therapy, hepcidin, and calcium channel blockers are being actively investigated. There is a need for the development of clinical guidelines in order to improve the management of this emerging complex disease.


Synthesis and evaluation of some new retinoids for cancer chemoprevention.
Many derivatives and analogs of the natural vitamin A compounds have been synthesized, and some of these retinoids are very active in various bioassays that may be predictive for cancer chemopreventive activity. Analogs have been synthesized in which parts of one, or more, of the three traditional subdivisions of the retinoid structure--the terminal polar group, the conjugated side chain, and the cyclohexenyl region--have been altered or replaced. In addition, structural alterations have produced compounds (arotinoids) in which two, or more, aromatic rings replace much of the classical retinoid structure. Further structural modifications that effected replacement of the entire retinoid structure by aromatic groups and certain linking groups have been made. Some of the arotinoids and further structural variants, which appear superficially to represent immense alteration of the classical retinoid structure, have potent activity in various retinoid bioassays. Four types of new classical retinoids (retinoylamino acids, bifunctional analogs, retinyl ethers, and 4-oxoretinoid derivatives) have been synthesized and evaluated in our laboratories. All-trans- and 13-cis-retinoyl alpha-amino acids are active in a hamster trachea organ culture assay and show much promise in potentiating immune responses. Retinoic acid analogs that have a bifunctional terminus bind to CRABP even though they have a bulky alpha-substituent, induce differentiation of F9 embryonal carcinoma cells, and have good activity in an immune-potentiation assay. A new retinyl either, retinyl propynyl ether, has chemopreventive activity against mammary carcinogenesis in vivo. Oral administration of this retinoid produces high and sustained concentrations in the rat mammary gland. New congeners of 4-oxoretinoic acid demonstrate high activity in a spectrum of bioassays.


Changes in bone mineral density and bone turnover markers in patients undergoing hematopoietic stem cell transplant.
Hematopoietic stem cell transplant (HSCT) is frequently complicated by endocrine abnormalities and loss of bone mass. This prospective study was conducted to evaluate the bone loss post-HSCT. A total of 50 patients was evaluated pretransplantation, and 25 had HSCT (17 males, 8 females; 19 allogenic, 6 autologous). Bone mineral density (BMD) and bone markers were measured at baseline, 3-6 months and 12 months. The mean age and body mass index were 25.1 ± 16.3 years and 19.4 ± 4.5 kg/m(2), respectively. There were 15 adults (60%), and 10 adolescents (40%). There was a significant decline in BMD from the baseline at total femur (-8.7%; P < 0.0001), femoral neck (-5.0%; P = 0.003), femoral trochanter (-6.0%; P = 0.001), and Ward's triangle (-9.9%; P < 0.0001) at 6 months posttransplantation. From the 6 months to 12 months, there was a significant improvement in BMD at above sites except at Ward's triangle. The decline in BMD was nonsignificant at the whole body (-0.3%, P = 0.748) and the lumbar spine (-2.7%, P = 0.130) at 6 months posttransplant. Younger patients with allogenic graft and steroid use are more likely to have significant loss of BMD at hip posttransplant. Serum osteocalcin decreased, and N-telopeptide increased at 3-6 months, which return to baseline at 1-year posttransplant. A significant bone loss is observed at 6 months in patients with post-HSCT. The bone loss occurs predominantly at cortical bone. There is recovery of bone mass at 12 months posttransplant except at Ward's triangle. Bone loss after HSCT is multifactorial.


Lipid Nanoparticles with Accessible Nickel as a Vaccine Delivery System for Single and Multiple His-tagged HIV Antigens.
Lipid-based nanoparticles (NPs) with a small amount of surface-chelated nickel (Ni-NPs) were developed to easily formulate the HIV his-tagged Tat protein, as well as to formulate and co-deliver two HIV antigens (his-p24 and his-Nef) on one particle. Female BALB/c mice were immunized by s.c. injection with his-Tat/Ni-NP formulation (1.5 μg Tat-his/mouse) and control formulations on day 0 and 14. The day 28 anti-Tat specific IgG titer with his-Tat/Ni-NP was significantly greater than that with Alum/his-Tat. Furthermore, splenocytes from his-Tat/Ni-NP immunized mice secreted significantly higher IFN-γ than those from mice immunized with Alum/his-Tat. Although Ni-NPs did not show better adjuvant activity than Tat-coated anionic NPs made with sodium dodecyl sulfate (SDS/NPs), they were less toxic than SDS/NPs. The initial results indicated that co-immunization of mice using his-p24/his-Nef/Ni-NP induced greater antibody response compared to using Alum/his-p24/his-Nef. Co-delivery of two antigens using Ni-NPs also increased the immunogenicity of individual antigens compared to delivery of a single antigen by Ni-NPs. In conclusion, Ni-NPs are an efficient delivery system for HIV vaccines including both single antigen delivery and multiple antigen co-delivery.


Reducing radiation exposure from computed tomography of the brain in children--report of a practical approach.
To reduce radiation exposure to paediatric neurosurgical patients from computed tomography (CT), a CT scanning protocol - lower radiation dose and selective scan segment (LDSS) protocol was used for CT brain at the authors' hospital. To evaluate the amount of reduction in radiation exposure by using this LDSS protocol compared to their usual protocol, the authors prospectively documented their findings. From May 2010 to June 2011, paediatric neurosurgical patients requiring CT brain, and when it was not a clinical emergency and there was baseline CT or MRI brain available, were evaluated for the LDSS protocol. The LDSS protocol used a lower tube current-time product and a shorter scan length to attain a lower total radiation dose. The CT scanning parameters of the patients' usual CT brain and LDSS CT were recorded and compared. Adverse events were also recorded. A total of 24 paediatric patients were included. Using the LDSS protocol, the effective doses were between 9% and 80% of the usual protocol and, in 20 patients, ≤50% of the usual protocol. The tube voltage was 120 kV. For patients below 10 years old, 100 mA s was adequate for the purposes of their CTs; in some patient categories, it was lower than 100 mA s. For patients aged 10 or above, 150 mA s was used. The scan length varied. Radiation exposure from CT brain in paediatric neurosurgical patients could be reduced by adopting a CT scanning protocol, which aimed dynamically at a lower tube current-time product and a shorter scan length than the usual settings at a hospital.


Relationships of herbivore feeding and plant flavonoids (Coleoptera: Chrysomelidae and Anacardiaceae: Rhus).
Eighteen leaf flavonoid compounds were isolated from several populations of Rhus tripartita from xeric habitats in Israel, ornamental Schinus terebinthifolius from Israel, and three species of Rhus (vulgaris, natalensis, tenuinervis) from mesic habitats in Kenya. Foodplant preference testing of Rhus-feeding Leaf Beetles (Blepharida sacra from Israel and B. marginalis and B. conradsi from Kenya) correlated well with the flavonoid composition of the different foodplants. Blepharida sacra and B. marginalis foodplant preferences demonstrated an herbivore sibling relationship but there is evidence, including foodplant ecology and distribution, that they are separate species. The herbivorefoodplant coevolution of the xeric B. sacra-R. tripartita is distinct from that of the mesic B. marginalis-R. vulgaris/natalensis, however, this study also indicates possible ancestral relationships between the herbivore species as well as between the plant species.


[A Monte-Carlo study for propensity score methods].
To evaluate the statistical property of the estimators on exposure effect and value of propensity score methods in practical use. Simulation data, propensity score methods (PSM) were conducted to assess bias and efficiency, under both with/without model misspecification conditions. Under model misspecification conditions, propensity score methods had better robustness than model based methods. For large and complicated data, propensity score methods had more flexibility in practical use than model based methods.


Temperament and sleep behaviors in infants and toddlers living in low-income homes.
Temperament is a dynamic trait that can be shaped by maturity and environmental experiences. In this study, we sought to determine whether and the extent to which temperament was predicted by sleeping behaviors in an understudied sample of primarily Black and White infants and toddlers living in low-income homes (N = 150). Sleeping behaviors were assessed at 15-19 months of age with caregiver report of the Tayside Children's Sleep Questionnaire. Temperament was examined as effortful control, negativity, and surgency with the Infant Behavior Questionnaire Very Short Form at 9-12 months of age and with the Early Childhood Behavior Questionnaire Short Form at 20-24 months of age. Covariates were maternal education, household income, and child sex and race. Continuous variables were standardized, then missing data from independent variables were multiply imputed in 20 datasets. Regression analyses showed that about 1 SD improvement in toddler sleep behaviors significantly predicted about 1/5 SD better toddler effortful control. However, sleep behaviors did not significantly predict toddler surgency or negative affect. This study shows that for a sample of infants and toddlers in low-income homes, how a child learns to regulate sleeping behaviors may influence the development of overall effortful control about six months later.


Resonance Raman and ligand binding studies of the oxygen-sensing signal transducer protein HemAT from Bacillus subtilis.
HemAT-Bs is a heme-containing signal transducer protein responsible for aerotaxis of Bacillus subtilis. The recombinant HemAT-Bs expressed in Escherichia coli was purified as the oxy form in which oxygen was bound to the ferrous heme. Oxygen binding and dissociation rate constants were determined to be k(on) = 32 microm(-1) s(-1) and k(off) = 23 s(-1), respectively, revealing that HemAT-Bs has a moderate oxygen affinity similar to that of sperm whale myoglobin (Mb). The rate constant for autoxidation at 37 degrees C was 0.06 h(-1), which is also close to that of Mb. Although the electronic absorption spectra of HemAT-Bs were similar to those of Mb, HemAT-Bs showed some unique characteristics in its resonance Raman spectra. Oxygen-bound HemAT-Bs gave the nu(Fe-O(2)) band at a noticeably low frequency (560 cm(-1)), which suggests a unique hydrogen bonding between a distal amino acid residue and the proximal atom of the bound oxygen molecule. Deoxy HemAT-Bs gave the nu(Fe-His) band at a higher frequency (225 cm(-1)) than those of ordinary His-coordinated deoxy heme proteins. CO-bound HemAT-Bs gave the nu(Fe-CO) and nu(C-O) bands at 494 and 1964 cm(-1), respectively, which fall on the same nu(C-O) versus nu(Fe-CO) correlation line as that of Mb. Based on these results, the structural and functional properties of HemAT-Bs are discussed.


Trophic effects of different enteral diets in the rat intestine.
The trophic effect of 4 commercial diets compared to chow was studied in 50 rats. The 4 commercial diets were: a free amino acid diet, a diet based on soy protein hydrolysate and two diets based on intact protein with and without extra fibre added. Bowel weight, mucosa weight and DNA content were measured after 7 days of treatment. The enteral formula based on a soy protein hydrolysate had a significant growth stimulating effect on the proximal small bowel compared to rat chow. In the ileum and in the colon all diets without fibre supplementation caused significantly lower bowel weights than rat chow.


Web-based dietary assessment and advice helps inflammatory bowel disease patients to improve their diet quality.
Time to evaluate diet quality and give dietary advice is limited in clinical IBD practice. The Eetscore is a web-based tool that assesses diet quality according to the Dutch dietary guidelines and provides personalised dietary advice. We aimed to assess diet quality of IBD patients using the Eetscore and to study changes in diet quality, health-related quality of life (HRQoL) and clinical disease activity over time. A prospective cohort study was performed in 195 adult IBD patients. Participants were invited to fill out questionnaires (Eetscore-FFQ, short IBDQ and p-HBI/p-SCCAI) at baseline and after 1 and 4 months. The Eetscore calculates diet quality based on 16 food components (10 points per component, total score 0-160; the higher the better) and provides dietary advice per component based on the assessment. At baseline, mean diet quality was 98±19. Diet quality was positively associated with age, female gender and level of education. Component scores were highest for red meat, wholegrain products, and sweetened beverages, and lowest for legumes, nuts, and processed meat. Over time, diet quality increased to 107±21 at 4 months (p<0.001). Each 10-point improvement in diet quality was associated with an increase in HRQoL (β=0.4 (95%CI 0.02; 0.7), p=0.04). Clinical disease activity did not change. In conclusion, diet quality of IBD patients significantly improved following personalised dietary advice of the Eetscore. Improvement of diet quality was associated with a slight improvement in HRQoL. The Eetscore is a practical and useful tool to monitor and support a healthy diet in IBD patients.


Prognostic value of the standardized uptake value maximum change calculated by dual-time-point (18)F-fluorodeoxyglucose positron emission tomography imaging in patients with advanced non-small-cell lung cancer.
The purpose of this study was to investigate the prognostic value of the standardized uptake value maximum (SUVmax) change calculated by dual-time-point (18)F-fluorodeoxyglucose positron emission tomography (PET) imaging in patients with advanced non-small-cell lung cancer (NSCLC). We conducted a retrospective review of 115 patients with advanced NSCLC who underwent pretreatment dual-time-point (18)F-fluorodeoxyglucose PET acquired at 1 and 2 hours after injection. The SUVmax from early images (SUVmax1) and SUVmax from delayed images (SUVmax2) were recorded and used to calculate the SUVmax changes, including the SUVmax increment (ΔSUVmax) and percent change of the SUVmax (%ΔSUVmax). Progression-free survival (PFS) and overall survival (OS) were determined by the Kaplan-Meier method and were compared with the studied PET parameters, and the clinicopathological prognostic factors in univariate analyses and multivariate analyses were constructed using Cox proportional hazards regression. One hundred and fifteen consecutive patients were reviewed, and the median follow-up time was 12.5 months. The estimated median PFS and OS were 3.8 and 9.6 months, respectively. In univariate analysis, SUVmax1, SUVmax2, ΔSUVmax, %ΔSUVmax, clinical stage, and Eastern Cooperative Oncology Group (ECOG) scores were significant prognostic factors for PFS. Similar results were significantly correlated with OS, except %ΔSUVmax. In multivariate analysis, ΔSUVmax and %ΔSUVmax were significant factors for PFS. On the other hand, ECOG scores were only identified as independent predictors of OS. Our results demonstrated the prognostic value of the SUVmax change in predicting the PFS of patients with advanced NSCLC. However, SUVmax change could not predict OS.


Assessment of thyroid cancer risk in more than 334,000 patients with inflammatory bowel disease: a case-control study and a meta-analysis.
Potential risk of thyroid cancer in patients with inflammatory bowel disease has not been well investigated. The aim of the study was to reveal the relationship between history of inflammatory bowel disease and risk of thyroid cancer. First, 1392 patients with inflammatory bowel disease and 1392 controls were included in a case-control study. All patients did not receive immunosuppressive therapy. A multivariate logistic regression analysis was adopted to determine the relationship between history of inflammatory bowel disease and risk of thyroid cancer. Second, a literature search was performed and eight articles were collected. Pooled odds ratios with 95% confidence intervals were reported for relevant risk estimates in fixed or random effect model. In the case-control study, thyroid cancer was more common in patients with inflammatory bowel disease than in controls (P = 0.032). After Bonferroni correction, association of thyroid cancer risk with history of total inflammatory bowel disease or its two subtypes was not found. In the meta-analysis, patients with total inflammatory bowel disease or ulcerative colitis showed an increased risk of thyroid cancer, but patients with Crohn's disease did not. Furthermore, inflammatory bowel disease patients with immunosuppressive therapy showed an increased risk of the cancer, but patients without immunosuppressive therapy did not have this finding. Risk of thyroid cancer probably elevates in patients with inflammatory bowel disease. Inflammatory bowel disease (particularly ulcerative colitis) itself and use of immunosuppressant might contribute to the development of the cancer.


Potential of lactic acid bacteria to reduce the growth ofFusarium culmorum in the malting process.
25 microorganisms were used for tested for their potential to reduce growth of toxigenic and gushing-activeFusarium culmorum strains during the malting process. Twelve isolates were found to substantially decrease the growth ofF. culmorum under the conditions applied.


Recognition of gasoline in fire debris using machine learning: Part I, application of random forest, gradient boosting, support vector machine, and naïve bayes.
The detection and identification of ignitable liquid (IL) residues in fire debris are two very challenging tasks in a fire investigation. To this day, the recognition of IL in fire debris includes the chemical analysis of the fire debris composition, followed by the examination and interpretation of the analysis result by a trained forensic examiner. Throughout the last decade, chemometrics and artificial intelligence have become increasingly important. In the present study, machine learning algorithms capable of recognizing gasoline residues in fire debris based on GC-MS data have been developed. Four methods, including random forest, gradient boosting, support vector machine, and naïve bayes are applied and used to classify fire debris samples into the two categories "with gasoline" or "without gasoline". A fifth method (logistic regression) did not converge due to well separated classes. A database comprising 360 measurements, including fire debris samples of real cases as well as fire debris samples spiked with known amounts of weathered gasoline (up to 99.6%), was available to train the machine learning algorithms (using 85% of the data) and to subsequently test the performance of the methods when classifying unknown samples (using 15% of the data). In general, the methods perform very well, as three of it succeeded to classify all test samples correctly without any false positive or false negative allocations. One (naïve bayes) was not trained enough to classify other (non-gasoline) IL correctly as "no gasoline". Furthermore, the random forest method reveals which chemical compounds are most relevant for the algorithm to classify the samples. In general, the presented approach is highly promising and could easily be extended or adapted to other types of IL. Similar to the neural network presented in the accompanying paper, such methods have the potential to serve as a fast screening technique for fire debris samples, thus supporting the forensic examiner by providing an additional independent opinion. Nonetheless, the definite identification of IL residues in fire debris always has to be accomplished by a forensic examiner.


Conventional heparin and semisynthetic heparin analogue (SSHA) alteration of blood coagulation after embolic occlusion of human renal circulation.
Blood coagulation and fibrinolytic variables were measured in a peripheral vein in a study of 21 consecutive patients before and after angio-embolization of renal carcinoma. The first ten patients received conventional heparin, 5000 IU twice daily, and the following eleven a semi-synthetic heparin analogue (SSHA), 50 mg twice daily, for 5 days. The first injection was given 2 hours before embolization and the last injection at least 12 hours before the last blood sampling. Both groups showed increased levels of FPA on day 5-7, indicating that the anticoagulant influence had ceased. F VII levels decreased only in the SSHA group from embolization to day 3, but were increased in both groups on day 5-7. Levels of thrombin-antithrombin complexes (TAT) were significantly increased in the heparin group 2 hours after embolization, indicating that thrombin activity had been formed. The corresponding TAT level in the SSHA group was not significantly increased. The differences could possibly indicate a different mechanism of action on blood coagulation of SSHA as compared with heparin, with involvement of extrinsic pathway and maybe by-passing antithrombin III inhibition.


A fluorescence spectroscopic method for rapid detection of bacterial endospores: Proof of concept.
Current spore detection methods rely on culture techniques, with limitations of time, efficiency, and sensitivity. The bacterial spore coat contains calcium dipicolinic acid (CaDPA) as a major constituent, which could serve as a biomarker for bacterial endospores. We report proof of concept for a rapid and sensitive technique for the detection of bacterial endospores by using ratiometric fluorescence-based sensors. This method is based on the detection of CaDPA, which enhances the luminescence of lanthanide ions when complexed with a semiconducting polymer. A CaDPA standard curve was generated at an excitation-emission wavelength (λ) of λ275-λ544 by using a spectrophotometer. The intensity was recorded after chelating semiconducting fluorescent polyfluorene (PFO) dots with terbium (lanthanide) ions, sensitized by different volumes of CaDPA (0.1 μM). The resultant standard curve showed a linear relationship (R2 = 0.98) in the experimental concentration range of 2.5 to 25 nM CaDPA, with corresponding intensity (arbitrary units) of 545 to 2,130. Endospores of the aerobic sporeformer Bacillus licheniformis ATCC 14580 were produced at 37°C for 15 d on brain heart infusion agar plates. The efficiency of sporulation was evaluated by spore staining and plating techniques. Total CaDPA content on spores was estimated after suspending decreasing concentrations of spores (logs 9.0 through 1.0 cfu/mL, at 1-log intervals) in HPLC-grade water (to serve as control) and skim milk samples. In HPLC-grade water, for higher spiking levels such as (mean ± SD) 9.2 ± 0.03, 8.4 ± 0.05, 7.1 ± 0.13, and 6.3 ± 0.02 logs, the corresponding mean CaDPA from the standard curve were 9.4, 7.2, 6.2, and 5.3 nM, respectively. For lower spiking levels of 4.2 ± 0.05, 3.1 ± 0.04, 2.0 ± 0.11, and 1.36 ± 0.09 logs, we observed mean CaDPA contents of 3.8, 3.3, 2.2, and 1.3 nM, respectively. For raw skim milk spiked with B. licheniformis ATCC 14580 spores, the mean CaDPA content on spores was approximately 2.5, 3.8, and 5.0 nM for spiking levels of 5.21, 6.39, and 9.47 log cfu/mL, respectively. Trials were conducted in replicates of 3 and means were compared. Trials conducted using HPLC-grade water showed a linear relationship for the CaDPA content of endospores and for endospore counts with the standard CaDPA concentration curve. For skim milk-spiked samples, we observed reduced fluorescence detection, which was 5 times lower than that of spiked samples in HPLC-grade water. The reduced fluorescence in skim milk could be due to the turbidity of the solution or to interference from proteins, amino acids, and other ions in milk. This study thus provides proof of concept for a potential application of this technique to rapidly detect bacterial endospores in the dairy and food industry. Further work is required to remove the interference of ionic components in milk to improve detection limits in milk and other dairy product matrices such as cheese, whey proteins, and reconstituted powders.


Neglected lymph nodal metastases in patients with renal cancer: when to extend the anatomical template of lymph node dissection during nephrectomy.
The role of lymph node dissection (LND) in patients with renal cell carcinoma (RCC) is still controversial. However, detecting lymph node invasion (LNI) is key due to prognostic implications and to identify patients who might benefit from adjuvant therapies such as adjuvant pembrolizumab. Out of 796 patients, 261 (33%) received eLND, of whom 62 (8%) for suspicious lymph node (LN) metastases at preoperative staging (cN1). eLND was divided in 3 anatomical areas: (1) hilar, (2) side-specific (pre-/para-aortic or pre-/para-caval) and (3) inter-aorto-caval nodes. Overall maximum LN diameter was measured by a dedicated radiologist for each patient. Multivariable logistic regression models (MVA) were tested for the effect of maximum LN diameter in predicting the presence of nodal metastases outside the anatomical area of cN1. LNI was confirmed in 50% of cN1, whilst only 13 out of 199 cN0 patients were pN1 at final histology (6.5%; p < 0.001). In a per-patient analysis, of 62 cN1 patients, 24% vs. 18% vs. 8% harboured pN1 disease only inside vs. in-outside vs. only outside the suspicious anatomical field of cN1 at preoperative CT/MRI scan. At MVA, increasing diameter of suspicious LNs was independently associated with risk of finding positive LNs outside the suspicious anatomical field (OR 1.05, 95%CI 1.02-1.11; p = 0.02). Roughly 50% of cN1 patients undergoing eLND will harbour LN metastases, also outside the suspicious radiological area, and maximum LNs diameter at preoperative imaging correlates with such risk. Thus, an eLND might be justified in patients with large suspicious LN metastases, to better stage this patient population and to improve postoperative treatment management.


Total dural calcification with secondary hyperparathyoidism: a rare entity.
A case of total, dense dural calcification in an adult female patient with secondary hyperparathroidism is presented. The often reported feature in the skull is that of a combination of osteopenia and osteosclerosis giving a granular bone texture described sometimes as "grains of sand" or "salt and pepper" appearance [6]. Extensive dural calcification with this condition has very rarely been reported [2, 3]. The purpose of this paper is to document the existence of the latter in a patient followed up for 19 years.


Modification by islet-activating protein of direct and indirect inhibitory actions of adenosine on rat atrial contraction in relation to cyclic nucleotide metabolism.
Adenosine decreased the force of contraction of isolated rat left atria (direct action) and inhibited the positive inotropic action of isoproterenol (indirect action). The direct action of adenosine was not accompanied by changes in cyclic nucleotide levels, while the isoproterenol-induced increase in cyclic AMP level was reduced by the nucleoside. Reserpine pretreatment and pindolol did not affect the direct action of adenosine. Both the direct and indirect actions of adenosine were enhanced by dipyridamole and dilazep and inhibited by 1-methyl-3-isobutylxanthine and theophylline. Among adenosine derivatives, N6-phenylisopropyladenosine was most potent, and 2'-deoxyadenosine was least effective in inducing the negative inotropic action in the absence and presence of isoproterenol. In atria pretreated with islet-activating protein (IAP) both the direct and indirect actions of adenosine, and the actions of N6-phenylisopropyladenosine were markedly attenuated compared with those in nontreated atria. The isoproterenol-induced increase in cyclic AMP level was not reduced by adenosine in IAP-treated atria. These results indicate that adenosine produces a dual action on rat atria: it decreases the force of contraction directly by a mechanism independent of cyclic AMP metabolism, and inhibits the positive inotropic action of isoproterenol by reducing the drug-induced accumulation of cyclic AMP. IAP attenuates both of these inhibitory actions of adenosine on rat atria.


Freezing of gait in Parkinson's disease: where are we now?
Freezing of gait (FOG) is defined as a brief, episodic absence or marked reduction of forward progression of the feet despite the intention to walk. It is one of the most debilitating motor symptoms in patients with Parkinson's disease (PD) as it may lead to falls and a loss of independence. The pathophysiology of FOG seems to differ from the cardinal features of PD and is still largely unknown. In the present paper, we review the studies that were performed since 2011 on methods to provoke and assess FOG and discuss new insights into behavioral and neural mechanisms underlying this clinical phenomenon. We conclude that most of the work reviewed confirms that gait pattern generation disturbances are central to FOG. The finding that FOG reflects a combined motor and cognitive de-automatization deficit, which may not be sufficiently offset by executive control, probably acts as parallel mechanism.


Histopathological discrepancy and variation of surgical management in mucinous ovarian cystadenoma and pseudomyxoma peritonei.
Mucinous cystadenoma occurs in 10-15% of all ovarian tumors. Diagnosis and treatment should be decided precisely as it has a chance to develop into pseudomyxoma peritonei (PMP). Management of PMP might be challenging especially when repeated surgery is needed. The first case, a 22-year-old lady with recurrent stomach enlargement for seven months. She had history of laparotomy surgery due to an ovarian tumor. Whole abdomen contrast CT scan showed a large cyst with mucinous fluid. We decided to do re-laparotomy and found a left ovarian cyst. Histological examination results confirm ovarian mucinous cystadenoma. The second case was, 55-year-old woman, with abdominal enlargement for six months. She had a history of laparotomy and chemotherapy due to pseudomyxoma peritonei. Post chemotherapy MRI showed persistent pseudomyxoma and two multilocular cysts from both adnexa. Debulking laparotomy was then conducted. We obtained 8 L of mucinous pseudomyxoma along with mucinous cyst from both ovaries. The final diagnosis concluded as a pseudomyxoma and we decide to close the follow-up of the patient. Pseudomyxoma is caused by the production of mucin originating from intra-abdominal organs. Open surgery should be prioritized when the mucinous cystadenoma is detected to do a complete peritoneum evaluation and avoid perioperatively ruptured mucinous neoplasm. Pseudomyxoma often needed repeated surgical treatment and may exhibit different surgical findings and different pathologies. Repeated surgery is logical and still no need for adjuvant chemotherapy in both cases. Accurate and precise diagnosis should be prioritized in order to prevent repeated surgery.


Value of image enhancement and injection of contrast medium for right ventricular volume determination by two-dimensional echocardiography in congenital heart disease.
To determine factors that influence the accuracy of echocardiographically estimated right ventricular volume and to improve the echocardiographic input information by applying image enhancement techniques, quantitative contrast echocardiography (4-chamber view) and biplane angiocardiography were performed in 23 children during routine diagnostic cardiac catheterization. Volumes calculated on the basis of unprocessed and processed echocardiographic cross sections (area-length method and sphere model) underestimated angiocardiographic volumes significantly (p less than 0.01), and more so in end-diastole (50.6%) than in end-systole (35.9%). Thus, ejection fraction was significantly (p less than 0.01) underestimated; mean values were 0.48 +/- 0.12 and 0.60 +/- 0.08, respectively. The best comparison between echocardiography and angiocardiography at end-diastole was achieved with the sphere model using image enhancement techniques and injection of contrast media, where y = 0.54x - 6.8, r = 0.97, sy.x = 7.3. Correlations, however, in which unprocessed echocardiograms were used showed only slightly less good correlations. With the 6 image-enhancement techniques, a more homogeneous structure of the image and a more distinct outline of the internal surface was achieved. The statistical error improved only slightly. The echocardiographic 4-chamber view allows right ventricular volume determination with an acceptable accuracy. Its underestimation is related to inadequate visualization of trabeculations and mainly to the models used. Application of image enhancement techniques allows easier outlining of the internal cavity surface. The advantage gained by the combination of contrast infection and image enhancement techniques does not warrant the routine central injection of available contrast material.


Refractory Acute Antibody Mediated Rejection in Liver Transplant After Desensitization of Preformed Donor Specific Antibody-Validity of Bortezomib and Everolimus: A Case Report.
Here, we report a case of living donor liver transplantation (LDLT) complicated with severe acute antibody-mediated rejection (aAMR), although desensitization was performed for preformed donor-specific anti-human leukocyte antigen antibody (DSA). LDLT was performed in a 59-year-old woman with alcoholic cirrhosis with a graft from her 60-year-old husband as a living donor. She had reproductive history of 4 gravidity and parity with her husband. Preoperative serologic studies showed positive complement-dependent cytotoxic crossmatch and anti-human leukocyte antigen-A26 antibody was identified as DSA. Desensitization for preformed DSA with rituximab and plasma exchange was performed before LDLT. We decided to perform LDLT using her husband right liver as living donor graft since the DSA mean fluoro-intensity was down to negative range. The immunosuppressive regimen was comprised with steroid and tacrolimus. However, the recipient developed acute cellular rejection on day 5 after LDLT, followed by severe aAMR. Re-administration of rituximab followed by 4 courses of plasma exchange failed to treat aAMR. The DSA mean fluoro-intensity was successfully suppressed after bortezomib was administered however impaired serologic liver function test and cholestasis were remained. The liver function test and cholestasis in the graft were improved after Everolimus was administered. The recipient was discharged on postoperative day 196. In conclusion, we report a case of LDLT who developed aAMR after desensitization of preformed DSA and was successfully treated with intensive therapy with bortezomib and everolimus.


Therapeutic education for empowerment and engagement in patients with Parkinson's disease: A non-pharmacological, interventional, multicentric, randomized controlled trial.
In 1997 the European Parkinson's Disease Associations launched the Charter for People with Parkinson's disease that stated the right of patients to be informed and trained on the disease, its course, and treatments available. To date, few data analyzed the effectiveness of education program on motor and non-motor symptoms of PD. The aim of this study was to evaluate the efficacy of an education program as it was a pharmacological treatment, thus choosing as the primary endpoint the change in daily OFF hours, the most widely used outcome in pharmaceutical clinical trials on PD patients with motor fluctuations. Secondary outcomes were change in motor and non-motor symptoms, quality of life and social functioning. The long-term efficacy of the education therapy was also evaluated by analyzing data collected at 12- and 24-weeks follow-up outpatient visits. One hundred and twenty advanced patients and their caregivers were assigned to the intervention or control group in a single-blind, multicentric, prospective, randomized study evaluating an education program structured in individual and group sessions over a 6-weeks period.At the end of study, the intervention group showed a significant reduction in daily OFF hours compared to control patients (-1.07 ± 0.78 vs. 0.09 ± 0.35, p < 0.0001) and a significant improvement was also reported in most secondary outcomes. Patients retained significant medication adherence and daily OFF hours reduction at 12- and 24-weeks follow-up. The results obtained demonstrated that education programs may translate in a notable improvement in motor fluctuations and non-motor symptoms in advanced PD patients.Clinical Trial Registration:Clinicaltrials.gov, identifier NCT04378127.


Passive signal propagation and membrane properties in median photoreceptors of the giant barnacle.
1. The light-induced electrical responses of barnacle photoreceptors spread decrementally along the cells' axons. The decay of the depolarizing and hyperpolarizing components of the visual signal was studied by recording intracellularly from single receptor axons of the median ocellus of the giant barnacle.2. The resistance of the photoreceptor neurone decreases markedly when the cell is depolarized with respect to its dark resting potential of -60 mV. This rectification results in differential attenuation of the depolarizing and hyperpolarizing components of the visual signal as they spread down the axon. Consequently, the visual signal entering the synaptic region is conspicuously distorted.3. Bathing the photoreceptor axons in sodium-free or calcium-free saline or in isotonic sucrose does not significantly affect the spread of the visual signal to the terminals. Thus the signal is not amplified by an ionic mechanism along the axon.4. Membrane characteristics of the photoreceptor for hyperpolarizing voltage changes were estimated from (a) the ratio of the amplitudes of the visual signals recorded simultaneously in the axon and in the soma, (b) the time constant, and (c) the input resistance of the cell. All three independent measurements are consistent with a length constant 1 to 2 times the total length of the cell (lambda = 10-18 mm) and an unusually high membrane resistivity of about 300 kOmega cm(2). This resistivity enables the receptor potential to spread passively to the terminal region.5. Electron microscopic examination of receptor axons reveals an investment of glial lamellae, but demonstrates neither unusual structures which would lead to a high apparent membrane resistivity, nor junctions between cells which would seal off the extracellular space. Thus the observed high resistivity appears to be an intrinsic property of the receptor membrane.


Protective effects of taurine on endothelial cells impaired by high glucose and oxidized low density lipoproteins.
Endothelial dysfunction, common to diabetes and cardiovascular diseases, is an early step in the development of atherosclerosis and diabetic angiopathies. Deficiencies of taurine have been related to diabetes and cardiovascular diseases. We investigated whether taurine provides protective action against endothelial dysfunction induced by hyperglycemia and/or oxidized low density lipoproteins (oxLDL). Quiescent human umbilical cord venous endothelial cells were exposed for 20 h to high glucose (35 mM) and/or oxLDL (60 microg/ml) alone and in presence of taurine (0.5-2.5 mg/ml). Apoptosis, caspase-3 activity, soluble(s) and cell surface expressions of vascular cellular (VCAM-1) and intercellular (ICAM-1) adhesion molecules were determined. Results are given as a percentage of the low glucose medium control. Apoptosis, VCAM-1 and ICAM-1 expressions were related to cell number. Hyperglycemia increased apoptosis to 162.5 +/- 19.2%, caspase-3 activity to 153.2 +/- 10.3%, cell-surface expression of VCAM-1 to 125.1 +/- 5.8%, the expression of ICAM-1 to 123.7 +/- 2.8% and sICAM-1 to 146.5 +/- 7.9%. Taurine (0.5-2.5 mg/ml) restored apoptosis, caspase-3 activity and expressions of VCAM-1 and ICAM-1. OxLDL (60 microg/ml) increased apoptosis to 114.8 +/- 3.1%; taurine (2.5 mg/ml) reduced this apoptosis to 40.5 +/- 4.1%. The combination of hyperglycemia and oxLDL increased apoptosis to 211.7 +/- 11.6%. This increase was normalized by taurine (2.5 mg/ml) to 97.9 +/- 12.8%. Taurine protects HUVECs from endothelial dysfunction induced by hyperglycemia through down-regulation of apoptosis and adhesion molecules. Counteracting the combination of oxLDL and hyperglycemia requires pharmacological concentrations of taurine.


ATP-dependent 6-phosphofructokinase from the hyperthermophilic bacterium Thermotoga maritima: characterization of an extremely thermophilic, allosterically regulated enzyme.
The ATP-dependent 6-phosphofructokinase (ATP-PFK) of the hyperthermophilic bacterium Thermotoga maritimawas purified 730-fold to homogeneity. The enzyme is a 140-kDa homotetramer composed of 34 kDa subunits. Kinetic constants were determined for all substrates in both reaction directions at pH 7 and at 75 degrees C. Rate dependence (forward reaction) on fructose 6-phosphate (F-6-P) showed sigmoidal kinetics with a half-maximal saturation constant ( S(0.5)) of 0.7 mM and a Hill coefficient of 2.2. The apparent K(m) for ATP was 0.2 mM and the apparent V(max) value was about 360 U/mg. The enzyme also catalyzed in vitro the reverse reaction with an apparent K(m) for fructose 1,6-bisphosphate and ADP of 7.6 mM and 1.4 mM, respectively, and an apparent V(max) of about 13 U/mg. Divalent cations were required for maximal activity; Mg(2+), which was most effective, could partially be replaced by Mn(2+) and Fe(2+). Enzyme activity was allosterically regulated by classical effectors of ATP-PFKs of Eukarya and Bacteria; it was activated by ADP and inhibited by PEP. The enzyme had a temperature optimum of 93 degrees C and showed a significant thermostability up to 100 degrees C. Using the N-terminal amino acid sequence of the subunit, the pfk gene coding for ATP-PFK was identified and functionally overexpressed in Escherichia coli. The purified recombinant ATP-PFK had identical kinetic and allosteric properties as the native enzyme purified from T. maritima. The deduced amino acid sequence showed high sequence similarity to members of the PFK-A family. In accordance with its allosteric properties, ATP-PFK of T. maritima contained the conserved allosteric effector-binding sites for ADP and PEP.


Microkeratome-assisted mushroom keratoplasty with minimal endothelial replacement.
To report the outcome of a new keratoplasty technique aimed at treating full-thickness opacities with minimal removal of recipient endothelium. Interventional case report. A deep central scar was removed using microkeratome-assisted mushroom-shaped keratoplasty, consisting of a large anterior stromal lamella (9.0-mm in diameter) and a small posterior button (5.0-mm in diameter) including deep stroma and endothelium. Complete suture removal was performed 3 months after surgery. Visual acuity, refraction, and computerized corneal topography were evaluated pre- and postoperatively. Best spectacle-corrected visual acuity improved from 20/60 to 20/20 at 6 months postoperatively. Postoperative refraction was -2.50-1.00 x 20 degrees. Microkeratome-assisted mushroom keratoplasty may offer visual and refractive advantages over conventional keratoplasty surgery. Because most of the recipient endothelium is preserved and may spread onto the posterior surface of the small donor button, graft decompensation secondary to immunologic rejection may be avoided with this technique.


Observation of non-linear biomass-capacitance correlations: reasons and implications for bioprocess control.
Electrical capacitance has been discussed as a real time measure for living biomass concentration in technical bioreactors such as brewery (fermentation) tanks. Commonly, a linear correlation between biomass concentration and capacitance is assumed. While following the growth and subsequent lipid formation of the yeast Arxula adeninivorans we observed non-linearity between biomass concentration and capacitance. Capacitance deviation from linearity coincided with incipient lipid formation and depended on the intracellular lipid content. As the extent of deviation between capacitance and biomass concentration was proportional to the lipid concentration, it was considered as a quantitative measure of intracellular product formation. The correlation between shifts in dielectric relaxation (summarized as characteristic frequency of the Cole-Cole equation) and lipid content could not be explained by interfacial polarization on the lipid droplets alone. However, the parameters of the Cole-Cole equation were found to be a clear indicator for different phases of growth and lipid production. Integrating all results in a redundancy analysis (RDA), we were able to accurately describe the formation of cellular lipid inclusions. Our measurements are thus potentially valuable as components of future bioprocess control strategies targeting intracellular products such as proteins or biopolyesters.


A Novel Semiconductor-Based Flow Cytometer with Enhanced Light-Scatter Sensitivity for the Analysis of Biological Nanoparticles.
The CytoFLEX is a novel semiconductor-based flow cytometer that utilizes avalanche photodiodes, wavelength-division multiplexing, enhanced optics, and diode lasers to maximize light capture and minimize optical and electronic noise. Due to an increasing interest in the use of extracellular vesicles (EVs) as disease biomarkers, and the growing desire to use flow cytometry for the analyses of biological nanoparticles, we assessed the light-scatter sensitivity of the CytoFLEX for small-particle detection. We found that the CytoFLEX can fully resolve 70 nm polystyrene and 98.6 nm silica beads by violet side scatter (VSSC). We further analyzed the detection limit for biological nanoparticles, including viruses and EVs, and show that the CytoFLEX can detect viruses down to 81 nm and EVs at least as small as 65 nm. Moreover, we could immunophenotype EV surface antigens, including directly in blood and plasma, demonstrating the double labeling of platelet EVs with CD61 and CD9, as well as triple labeling with CD81 for an EV subpopulation in one donor. In order to assess the refractive indices (RIs) of the viruses and EVs, we devised a new method to inversely calculate the RIs using the intensity vs. size data together with Mie-theory scatter efficiencies scaled to reference-particle measurements. Each of the viruses tested had an equivalent RI, approximately 1.47 at 405 nm, which suggests that flow cytometry can be more broadly used to easily determine virus sizes. We also found that the RIs of EVs increase as the particle diameters decrease below 150 nm, increasing from 1.37 for 200 nm EVs up to 1.61 for 65 nm EVs, expanding the lower range of EVs that can be detected by light scatter. Overall, we demonstrate that the CytoFLEX has an unprecedented level of sensitivity compared to conventional flow cytometers. Accordingly, the CytoFLEX can be of great benefit to virology and EV research, and will help to expand the use of flow cytometry for minimally invasive liquid biopsies by allowing for the direct analysis of antigen expression on biological nanoparticles within patient samples, including blood, plasma, urine and bronchoalveolar lavages.


Antihypertensive effects of L-tryptophan are not mediated by brain serotonin.
L-Tryptophan produces a significant antihypertensive effect in spontaneously hypertensive rats while D-tryptophan does not change blood pressure. Both isomers of tryptophan significantly increase brain serotonin to the same extent at a time when the antihypertensive effect of L-tryptophan is maximal. Thus, the antihypertensive effects of L-tryptophan do not appear to be mediated by brain serotonin.


Additions to Phaeosphaeriaceae (Pleosporales): Elongaticollum gen. nov., Ophiosphaerella taiwanensis sp. nov., Phaeosphaeriopsis beaucarneae sp. nov. and a new host record of Neosetophoma poaceicola from Musaceae.
A novel ascomycetous genus, Elongaticollum, occurring on leaf litter of Hedychium coronarium (Zingiberaceae) in Taiwan, is described and illustrated. Elongaticollum is characterized by dark brown to black, superficial, obpyriform, pycnidial conidiomata with a distinct elongate neck, and oval to oblong, hyaline, aseptate conidia. Phylogenetic analyses (maximum likelihood, maximum parsimony and Bayesian) of combined ITS, LSU, SSU and tef1-α sequence data revealed Elongaticollum as a distinct genus within the family Phaeosphaeriaceae with high statistical support. In addition, Ophiosphaerella taiwanensis and Phaeosphaeriopsis beaucarneae are described as new species from dead leaves of Agave tequilana and Beaucarnea recurvata (Asparagaceae), respectively. Neosetophoma poaceicola is reported as a new host record from dead leaves of Musa acuminata (Musaceae). Newly described taxa are compared with other similar species and comprehensive descriptions and micrographs are provided.


CGRP receptor activity in mice with global expression of human receptor activity modifying protein 1.
CGRP is a potent vasodilator and nociceptive neuropeptide linked to migraine. CGRP receptors are heterodimers of receptor activity modifying protein 1 (RAMP1) and either calcitonin receptor-like receptor (CLR; forms canonical CGRP receptor) or calcitonin receptor (CT receptor; forms AMY1 receptor). The goal of this study was to test whether transgenic mice globally expressing human RAMP1 have increased CGRP receptor activity and whether the receptors are sensitive to human selective antagonist telcagepant. cAMP production was measured in primary cultures of aortic smooth muscle and trigeminal ganglia neurons from global hRAMP1 mice and non-transgenic littermates. Functional activity and inhibition were compared with clonal cell lines expressing combinations of CLR or CT receptors with RAMP1. Cultured smooth muscle from global hRAMP1 mice had a 10-fold greater CGRP-induced cAMP maximal response (Rmax) than non-transgenic littermates, with similar EC50 s. In contrast, cultured trigeminal ganglia from global hRAMP1 mice had a 40-fold leftward shift of the EC50 , with similar Rmax values as littermates. In both hRAMP1 cultures, telcagepant blocked CGRP-induced cAMP production, but was not effective in non-transgenic cultures. IC50 values were closer to those observed for CT receptor/hRAMP1 than CLR/hRAMP1 in clonal cell lines. Overexpression of hRAMP1 increases CGRP signalling by changing the maximal response or ligand sensitivity, depending on tissue type. Furthermore, telcagepant inhibited transgenic hRAMP1 CGRP receptors, but the degree of inhibition suggests that the transgenic mice are only partially humanized or both canonical CGRP and AMY1 receptors are functional in trigeminal ganglia neurons and vascular smooth muscle.


A Linear Diffusion Model of Adsorption Kinetics at Fluid/Fluid Interfaces.
The paper presents a new model for kinetically controlled adsorption at the fluid/fluid interface. The main purpose of the presented approach is to relate easy to estimate bulk surfactant concentration with Gibbs surface excess. Two adsorption isotherms are involved in the new model development: Frumkin and Szyszkowski isotherms. Additionally the Johannsen time profile of concentration in the adsorption layer is assumed and estimated in the model derivation. The proposed approach assumes the near interface, adsorptive layer which is described based on Fick's transient diffusion law. The solution to the model contains the estimation of effective diffusivities with adsorptive layer thickness as well. The experimental results of toluene/water + sodium dodecyl sulfate are presented and used for model verification.


The first ADC bearing the ferroptosis inducer RSL3 as a payload with conservation of the fragile electrophilic warhead.
The iron-dependent, non-apoptotic cell death, known as ferroptosis is an emerging strategy for the development of anticancer drugs. RSL3 was identified as an activator of ferroptosis through the inhibition of the glutathione peroxidase 4 (GPX4) which plays a crucial role in the cellular lipid oxidative stress. RSL3 is characterized by the presence of an electrophilic chloroacetyl moiety, namely warhead which covalently bonds to the catalytic and nucleophilic selenocysteine residue (Sec46) of GPX4. Like the major ferroptosis inducers, RSL3 suffers from lack of selectivity toward tumor cells. In this study, we report the first synthesis of an antibody-drug conjugate (ADC) containing RSL3 fragment and trastuzumab with the aim to deliver the agent selectively to tumors. The synthesis uses a judiciously chosen strategy to preserve the vital but fragile warhead. Full characterization of the ADC was accomplished, demonstrating the generation of a homogeneous DAR 8 conjugate. The robustness of the synthesis was successfully applied to another ADC associating the anti-CD74 mAb milatuzumab. The ADC induces ferroptotic cell death through reactive oxygen species accumulation and increases the activity of doxorubicin. The ADC associating trastuzumab and RSL3 may therefore offer potential applications in vectorized therapy alone or in combination with conventional chemotherapies.


Pelagic and coastal green turtles (Chelonia mydas) experience differences in chemical exposure and effect.
Green turtles foraging in coastal areas are exposed to land-based chemical pollutants that accumulate in the habitats to which they show high site fidelity. However, prior to coastal recruitment, they may be exposed to a different range of chemical threats. The recent development of species-specific in vitro bioassays for marine turtles allows for an effect-based assessment of toxicological endpoints. Blood was collected from green turtles of two life-stages, 'recent recruits' and 'coastal residents', in Hervey Bay and Moreton Bay. Organic contaminants were extracted from blood using the QuEChERS method, and cytotoxicity of the extracts measured in green turtle skin cells. Although not statistically significant, extracts from 'coastal residents' exhibited greater mean toxicity compared to 'recent recruits', possibly indicative of increased chemical accumulation from coastal habitat exposure. The bioassay results also indicated that turtles foraging in Hervey Bay are at greater risk of chemical exposure than those foraging in Moreton Bay.


Determinants of prevalence, acquisition, and persistence of human papillomavirus in healthy Mexican military men.
Human papillomavirus (HPV) infection is sexually transmitted, but the nature of the infection in males is poorly understood. We sought to identify determinants of HPV infection, acquisition, and persistence in 1,030 healthy military men in Mexico. From July 2000 to July 2003, trained interviewers administered a questionnaire, conducted a genital examination, and collected samples. The presence of multiple HPV types in genital cells from the urethra, urethral meatus, scrotum, penile shaft, and coronal sulcus was evaluated. At baseline 1,030 participants and after 1-year follow-up 336 individuals were sampled using a highly sensitive DNA reverse blot strip assay. HPV prevalence was 44.6%; infection with high-risk types was observed in 34.8% participants and 51.1% were multiply infected. After 1-year follow-up, 165 men remained free of HPV, 68 cleared their infection, 45 acquired one, and 37 remained infected with the same HPV type. The period prevalence was 50.9%, the incidence rate was 17.9/1,000 men-months [95% confidence interval (95% CI), 13.0-23.9], clearance was 54%, and persistence was 29.4%. At baseline, the number of partners before age 20 years, a history of a sexually transmitted disease, and the presence of condilomas significantly increased the association with HPV infection. Having anal intercourse with males was associated with the risk of acquiring a HPV infection (odds ratio, 5.2; 95% CI, 1.2-23). The odds ratio for persistent infection was 0.10 (95% CI, 0-0.87) in men who reported being circumcised compared with those who did not. High-risk sexual behavior increases the risk of HPV infection in males, whereas circumcision may lower the risk of persistence.


Dual A1 and A2A adenosine receptor antagonists, methoxy substituted 2-benzylidene-1-indanone, suppresses intestinal postprandial glucose and attenuates hyperglycaemia in fructose-streptozotocin diabetic rats.
Recent research suggests that adenosine receptors (ARs) influence many of the metabolic abnormalities associated with diabetes. A non-xanthine benzylidene indanone derivative 2-(3,4-dihydroxybenzylidene)-4-methoxy-2,3-dihydro-1 H-inden-1-one (2-BI), has shown to exhibit higher affinity at A1/A2A ARs compared to caffeine. Due to its structural similarity to caffeine, and the established antidiabetic effects of caffeine, the current study was initiated to explore the possible antidiabetic effect of 2-BI. The study was designed to assess the antidiabetic effects of several A1 and/or A2A AR antagonists, via intestinal glucose absorption and glucose-lowering effects in fructose-streptozotocin (STZ) induced diabetic rats. Six-week-old male Sprague-Dawley rats were induced with diabetes via fructose and streptozotocin. Rats were treated for 4 weeks with AR antagonists, metformin and pioglitazone, respectively. Non-fasting blood glucose (NFBG) was determined weekly and the oral glucose tolerance test (OGTT) was conducted at the end of the intervention period. Dual A1/A2A AR antagonists (caffeine and 2-BI) decreased glucose absorption in the intestinal membrane significantly (p < 0.01), while the selective A2A AR antagonist (Istradefylline), showed the highest significant (p < 0.001) reduction in intestinal glucose absorption. The selective A1 antagonist (DPCPX) had the least significant (p < 0.05) reduction in glucose absorption. Similarly, dual A1/A2A AR antagonists and selective A2A AR antagonists significantly reduced non-fast blood glucose and improved glucose tolerance in diabetic rats from the first week of the treatment. Conversely, the selective A1 AR antagonist did not reduce non-fast blood glucose significantly until the 4th week of treatment. 2-BI, caffeine and istradefylline compared well with standard antidiabetic treatments, metformin and pioglitazone, and in some cases performed even better. 2-BI exhibited good antidiabetic activity by reducing intestinal postprandial glucose absorption and improving glucose tolerance in a diabetic animal model. The dual antagonism of A1/A2A ARs presents a positive synergism that could provide a new possibility for the treatment of diabetes.


The biosynthesis of the histidine-rich protein of Plasmodium lophurae and the cloning of its gene in Escherichia coli.
Uninucleate trophozoites and schizonts of Plasmodium lophurae were labeled metabolically with [3H]proline. Analysis of labeled parasites indicated that the histidine-rich protein (HRP) was the major polypeptide synthesized by both developmental stages; in trophozoites it represented a larger proportion of total labeled polypeptides. Polyadenylated RNA was prepared from trophozoites and translated in a rabbit reticulocyte lysate system in the presence of [3H]histidine. As compared to the approx. 50 kDa mature protein, in the cell-free system HRP was synthesized as an approx. 58 kDa precursor polypeptide. Size-selected, polyadenylated RNA was used to construct a complementary double-stranded DNA library in pBR322 and plasmids containing HRP sequences were identified by hybridization with a synthetic oligonucleotide probe.


Evaluation of the non-randomness of protein compositions.
A method is described for assessing the non-randomness of protein compositions, based on the chi-squared statistic for the differences between the observed numbers of residues of each type and the numbers expected for a random distribution of codons. The analysis indicates that changes in at least 30% of the residues in natural proteins are selected against.


Updating "Intensity dataset acquired through laser scanning of lunar and Martian soil simulants" with additional lunar soil simulants specimens.
Updated dataset based on the laser scanning of three specimens of lunar soil simulants representing highland regions (LHS-1, AGK-2010, CHENOBI) and three specimens of lunar soil simulants representing mare regions (LMS-1, JSC-1A, OPRL2N). In previous studies, only a specimen of ilmenite was used as a lunar soil simulants. Measurements were executed using phase-shift and time-of-fly terrestrial laser scanners (Z + F IMAGER 5016, FARO Focus3D, and Leica ScanStation C10). The stored data contain radiometric information of point clouds from the measurement of lunar soil simulants. The data provide information on the effect of different types of simulants (mare and highland) on the absorption and dispersion of the laser beam. In overall, the dataset presented in this work can be used in investigation of the application of the laser technology in the measurement of lunar structures made of mare and highlands lunar regolith. Lunar highlands and mare regions are characterized by different physicochemical parameters. Therefore, both types of soil scatter the laser beam differently, what affects the range and accuracy of the laser measurements.


The treatment of herpes encephalitis.
The advent of acyclovir has accentuated the problem of diagnosis and particularly the need for brain biopsy in patients with suspected herpes simplex encephalitis. U.S.A. and U.K. practices are contrasted and the current U.K. study of acyclovir in herpes simplex encephalitis is mentioned in which retrospective and relatively non-invasive methods of diagnosis are employed.


Angiopeptin, a somatostatin-14 analogue, decreases adhesiveness of rat leukocytes to unstimulated and IL-1 beta-activated rat heart endothelial cells.
We have previously demonstrated that somatostatin-14 and its octapeptide analogue, angiopeptin, decrease the ability of rat heart endothelial cells to bind leukocytes [Leszczynski, et al., Reg. Pept. 43 (1993) 131-140]. Here, we examined whether exposure of leukocytes to angiopeptin modifies their adhesiveness to the unstimulated and to IL-1 beta-activated endothelium. Monolayers of unstimulated endothelial cells bind 274 +/- 12 leukocytes/mm2. Exposure of leukocytes for 1, 4 and 24 hours to angiopeptin (1 microM) reduced significantly (p < 0.05) adhesion of leukocytes from 274 +/- 12 to 188 +/- 10, 185 +/- 8 and 172 +/- 3 cells/mm2, respectively. Stimulation of endothelial cells with Il-1 beta (100 U/ml) for 24 hours increased endothelial adhesiveness from 274 +/- 12 to 381 +/- 17 adhering leukocytes/mm2. Exposure of leukocytes for 1, 4 and 24 hours to angiopeptin (1 microM) reduced significantly (p < 0.05) binding of leukocytes to IL-1 beta-activated endothelium from 381 +/- 17 to 237 +/- 8, 254 +/- 11 and 248 +/- 13 cells/mm2, respectively. Angiopeptin had no effect on the expression of lymphocyte function-associated molecule-1 (LFA-1; CD11a/CD18) by leukocytes, as assessed by flow cytometry. This suggests that angiopeptin modulates adhesive properties of leukocytes by (1) altering the expression of other than LFA-1 adhesion molecule(s) and/or (2) modulating the affinity of adhesion molecule(s) expressed by leukocytes. In conclusion, our results demonstrate that angiopeptin reduces leukocyte adhesiveness to unstimulated and to IL-1 beta-activated endothelium. It suggests that angiopeptin may suppress immune response via modulation of the leukocyte-endothelial interaction.


Spatiotemporal characteristics of agricultural nitrogen and phosphorus emissions to water and its source identification: A case in Bamen Bay,China.
The spatiotemporal characteristics and sources identification of agricultural nitrogen (N) and phosphorus (P) emissions to the gulf are rarely reported in tropical regions of China, mainly due to the lack of local reliable data and quantitative tools for spatiotemporal changes. In this study, we constructed a high-resolution NUFER (NUtrient Flow in food chains, Environment and Resources use) model based on geology, meteorology, land use data, statistical data, and field investigation to quantify the spatiotemporal characteristics and sources of N and P emissions. Bamen Bay (BMB), a bay with a mangrove national wetland Park in the Hainan Island, China, was chosen as a case study. The results showed that agricultural N emission to water in 2018 increased fivefold compared to 1990. Leaching was the main method of agricultural N emission and was mainly distributed in farms in the west and north. The contribution of N emission from crop system to water increased 20.3% in 28 years. Poultry and fruits have contributed the most to N output, and the trend is continuing. P emission to water increased sevenfold compared 1990. The contribution of P emission from animal system to water increased from 86.8% in 1990 to 90.1% in 2018 due to low removal rate of livestock manure. P emission was mainly via direct discharge of manure, mainly distributed in livestock breeding sites near the bay. Poultry has consistently contributed the most to P output in 28 years, accounting for 49.1% in 2018. Fertilizers and fodder were the largest sources of N and P. The average N and P loss rates of BMB were 5.32 t km2 yr-1 and 0.26 t km2 yr-1. The future agricultural transformation is essential, and it is necessary to reduce the application of N fertilizer and increase the removal rate of livestock manure. These results can provide reference for other typical agricultural pollution bays in exploring the spatiotemporal characteristics of N and P emissions to water and the identification of agricultural sources.


[Diagnostics of cardiac amyloidosis].
The early diagnosis of cardiac amyloidosis is decisive for the success of treatment of affected persons. The thorough clinical investigation of the patient should be followed by appropriate diagnostics using modern procedures. The main symptoms are dyspnea, loss of performance and edema and in later stages cardiac arrhythmias in the form of atrioventricular conduction disturbances and atrial fibrillation but ventricular arrhythmias occur more rarely. During heart failure due to cardiac amyloidosis an increase of cardiac enzymes frequently occurs (e.g., creatine kinase, troponin, N‑terminal pro-brain natriuretic peptide), which can be included in the risk stratification and treatment monitoring, taking certain limitations into consideration. The investigation of light chains in serum and/or urine should be carried out immediately, as soon as there is a clinical and echocardiographic suspicion of cardiac amyloidosis. Subsequently, either cardiac magnetic resonance imaging (MRI) or bone scintigraphy should be carried out, depending on the locally available options. Depending on the results of these two imaging procedures, a decision must be made as to whether further diagnostic steps (e.g., endomyocardial biopsy) are necessary. In the last decade bone scintigraphy has proven to be a blessing for the diagnostics of cardiac amyloidosis but many partial aspects and limitations necessitate special and careful consideration. A Perugini score of 2 or 3 is initially "indicative" of cardiac amyloidosis but not yet "confirmative" for a specific subtype. Only after an additional negative result of the light chain determination, can the diagnosis of ATTR amyloidosis be noninvasively made. Cardiac amyloidosis shows a particularly characteristic contrast enhancement in cardiac MRI, which mostly begins in the inner (subendocardial) layers of the basal left ventricular (LV) wall and frequently appears to be circular in the cross-sectional view of the left ventricle. Supplementary T1 and extracellular volume fraction mapping results, which are shown as color-coded maps, enable the rapid and elegant assessment of the myocardial structure and the extent of amyloid deposition. An additional investigation of the TTR gene is recommended in the case of ATTR amyloidosis for a differentiation between hereditary and acquired ATTR, as from this, further therapeutic consequences can be derived.


Exogenous administration of mesenchymal stem cells ameliorates dextran sulfate sodium-induced colitis via anti-inflammatory action in damaged tissue in rats.
Mesenchymal stem cells (MSCs) may modulate inflammatory responses resulting in improvement in inflammatory diseases, as well as tissue regeneration via cellular differentiation. We examined the therapeutic effects of exogenously administered MSCs in dextran sulfate sodium (DSS)-induced colitis in rats. Experimental colitis was produced in inbred male Lewis rats by administration of 4% DSS in drinking water for 7 days. MSCs (5x10(6) cells) which were isolated from whole marrow cells and cultured in an optimal medium for MSC outgrowth were administered to the treated rats via the tail vein on days 0, 2, and 4. On day 7, we evaluated colon length, histological changes, and colonic various mRNA expressions by RT-PCR. Localization of MSCs was evaluated using a green-fluorescent cell linker dye. To evaluate the anti-inflammatory action of MSCs, we assayed LPS-induced TNF-alpha secretion in a co-culture of MSCs and monocytes (THP-1 cells) using ELISA. MSCs reduced in bloody stools, weight loss, colon shortening, and microscopic injuries. In the rectum of MSCs-treated rats, mRNA expression of TNF-alpha, IL-1beta, and COX-2 decreased to 40, 15, and 15% of their respective control levels. MSCs significantly suppressed mRNA expression of VEGF, HGF, and b-FGF to 40, 25, and 25% of their respective control levels. Green-fluorescent-labeled MSCs were found only within the lamina propria in inflamed regions. LPS-induced TNF-alpha secretion by THP-1 cells was significantly suppressed by co-culture with MSCs dose-dependently. We conclude that exogenous MSCs accumulated in inflamed tissues and ameliorated DSS-induced colitis via a local anti-inflammatory action.


A case-report of the unprovoked thrombotic event in a patient with thymoma and severe FVII deficiency.
Factor VII deficiency is a rare bleeding disorder caused by a deficiency of clotting factor VII. However, there have been some case reports of venous thrombosis in patients with factor VII deficiency, especially underlying the prothrombotic risk factors exposure. Patients with factor VII deficiency require special considerations before undergoing surgery to minimize the risk of bleeding or thrombogenesis. Here, we described a patient with early-stage thymoma and severe factor VII deficiency who experienced an unprovoked thrombotic episode before thymectomy and a fatal thrombotic event after surgery. By adopting gene screening, a reported homozygous F7 mutation (p.His408Gln) and a novel heterozygous PROS1 mutation (p.Pro147Ala) were identified. The former resulted in severe factor VII deficiency but did not protect against thrombosis, and the latter was correlated with normal expression and cofactor activities of protein S through the thrombin generation test. The perioperative infusion of recombinant factor VII concentrate and the absence of antithrombotic prophylaxis may collectively contribute to her fatal thrombotic event after surgery. For the patients with severe factor VII deficiency undergoing surgery, uniform replacement therapy may not be recommended, and antithrombotic prophylaxis should be used in the case with thrombotic history to minimize the risk of bleeding and thrombogenesis.


Intracytoplasmic sperm injection allows normal pregnancy rates for males 40 with low hypoosmotic swelling test scores even when complicated by very low motility percentage.
To determine if the additional burden of low percentage motility reduces the chance that sperm with low hypoosmotic swelling (HOS) test scores will achieve a pregnancy following in vitro fertilization (IVF) with intracytoplasmic sperm injections (ICSI). Couples undergoing IVF-embryo transfer (ET) and ICSI for low HOS tests (< 50%) were retrospectively identified. The percentage motility was divided into deciles. Pregnancy rates were determined according to the deciles of motility. No differences in clinical or live delivered pregnancy rates per transfer were found in even the very lowest percent motility category. The added complicating factor of low percentage motility added to sperm with low HOS test scores does not reduce the effectiveness of IVF with ICSI.


Association of main meal frequency and skipping with metabolic syndrome in Korean adults: a cross-sectional study.
Reduced meal frequency can increase the risk of metabolic syndrome (MetS). However, limited studies have examined the association between meal frequency and skipping meals with MetS. This study aims to analyze the association between main meal frequency and meal skipping with MetS in Korean adults aged ≥ 19 years. In this study, we included data from 22,699 Korean adult participants from the 2016-2020 Korea National Health and Nutrition Examination Survey (KNHANES). The 24-h dietary recall method was used to classify the participants into three groups based on main meal frequency (one, two, or three meals per day) and seven groups based on the type of main meal they skipped. Multivariable logistic regression analysis was conducted to determine the association between main meal frequency and the types of main meals skipped with the odds of MetS and its associated components. Appropriate estimates were accounted for using sampling weights, stratification, and clustering. The prevalence of MetS in the study population was 33.8%. The average age of the participants was 47.2 years with 42.6% being men, and 57.4% being women. Men who consumed two meals per day had higher odds of MetS than those who consumed three meals per day (odds ratio [OR] 1.16, 95% confidence interval [CI] 1.01-1.33). Women who consumed two meals per day, and skipped breakfast had increased odds of having elevated fasting blood glucose levels (OR 1.18, 95% CI 1.02-1.35), and elevated triglycerides (OR 1.19, 95% CI 1.02-1.39). However, women who skipped dinner had reduced odds of having elevated fasting blood glucose levels (OR 0.74, 95% CI 0.58-0.94). Our findings suggest that meal frequency and the type of main meal skipped may be associated with MetS and emphasize the importance of consuming breakfast to prevent MetS.


Transition Metal/Lewis Acid Catalyzed Reactions of Zerumbone for Diverse Molecular Motifs.
Zerumbone is a naturally occurring humulene type sesquiterpene, isolated from the rhizomes of Zingiber zerumbet (L.) Smith with excellent therapeutic potential and is recognized as a valuable synthon for the construction of diverse array of natural product motifs. In this review, we intended to highlight our achievements in utilizing abundant natural product zerumbone and its derivatives for the development of pharmacologically relevant molecular scaffolds. We provided an account of the transition-metal catalyzed 1,4-conjugate addition reactions of zerumbone and its derivatives along with palladium-catalyzed cross-couplings, transition metal-based Lewis acid promoted interrupted Nazarov cyclisation reaction with substituted indoles and transannular cyclizations, photo-induced transformations of zerumbone and its epoxide.


Coexistent neuroblastoma and Hirschsprung's disease--another manifestation of the neurocristopathy?
Abnormalities of neural crest development may be manifested in a wide variety of ways ranging from von Recklinghausen's disease to Sipple's syndrome -- pheochromocytoma and medullary thyroid carcinoma. We report here yet another manifestation -- coexistent multiple neuroblastomas and Hirschsprung's disease in a newborn. In a review of the English literature we have found the combination of neuroblastoma and Hirschsprung's disease recorded only once previously [2]. However, the relationship between the two diseases was not discussed. The two diseases may be at the extremes of a spectrum of neural crest development pathology, i.e., Hirschsprung's representing a deficiency and neuroblastoma representing malignant proliferation of cells of neural crest origin.


How Heterogeneity in Glucokinase and Gap-Junction Coupling Determines the Islet [Ca2+] Response.
Understanding how cell subpopulations in a tissue impact overall system function is challenging. There is extensive heterogeneity among insulin-secreting β-cells within islets of Langerhans, including their insulin secretory response and gene expression profile, and this heterogeneity can be altered in diabetes. Several studies have identified variations in nutrient sensing between β-cells, including glucokinase (GK) levels, mitochondrial function, or expression of genes important for glucose metabolism. Subpopulations of β-cells with defined electrical properties can disproportionately influence islet-wide free-calcium activity ([Ca2+]) and insulin secretion via gap-junction electrical coupling. However, it is poorly understood how subpopulations of β-cells with altered glucose metabolism may impact islet function. To address this, we utilized a multicellular computational model of the islet in which a population of cells deficient in GK activity and glucose metabolism was imposed on the islet or in which β-cells were heterogeneous in glucose metabolism and GK kinetics were altered. This included simulating GK gene (GCK) mutations that cause monogenic diabetes. We combined these approaches with experimental models in which gck was genetically deleted in a population of cells or GK was pharmacologically inhibited. In each case, we modulated gap-junction electrical coupling. Both the simulated islet and the experimental system required 30-50% of the cells to have near-normal glucose metabolism, fewer than cells with normal KATP conductance. Below this number, the islet lacked any glucose-stimulated [Ca2+] elevations. In the absence of electrical coupling, the change in [Ca2+] was more gradual. As such, electrical coupling allows a large minority of cells with normal glucose metabolism to promote glucose-stimulated [Ca2+]. If insufficient numbers of cells are present, which we predict can be caused by a subset of GCK mutations that cause monogenic diabetes, electrical coupling exacerbates [Ca2+] suppression. This demonstrates precisely how metabolically heterogeneous β-cell populations interact to impact islet function.


Scalable and high-throughput production of an injectable platelet-rich plasma (PRP)/cell-laden microcarrier/hydrogel composite system for hair follicle tissue engineering.
Tissue engineering of hair follicles (HFs) has enormous potential for hair loss treatment. However, certain challenges remain, including weakening of the dermal papilla cell (DPC) viability, proliferation, and HF inducibility, as well as the associated inefficient and tedious preparation process required to generate extracellular matrix (ECM)-mimicking substrates for biomolecules or cells. Herein, we utilized gelatin methacryloyl (GelMA) and chitosan hydrogels to prepare scalable, monodispersed, and diameter-controllable interpenetrating network GelMA/chitosan-microcarriers (IGMs) loaded with platelet-rich plasma (PRP) and seeded with DPCs, on a high-throughput microfluidic chip. The ECM-mimicking hydrogels used for IGMs exhibited surface nano-topography and high porosity. Mass production of IGMs with distinct and precise diameters was achieved by adjusting the oil and aqueous phase flow rate ratio. Moreover, IGMs exhibited appropriate swelling and sustained growth factor release to facilitate a relatively long hair growth phase. DPCs seeded on PRP-loaded IGMs exhibited good viability (> 90%), adhesion, spreading, and proliferative properties (1.2-fold greater than control group). Importantly, PRP-loaded IGMs presented a higher hair inducibility of DPCs in vitro compared to the control and IGMs group (p < 0.05). Furthermore, DPC/PRP-laden IGMs were effectively mixed with epidermal cell (EPC)-laden GelMA to form a PRP-loaded DPC/EPC co-cultured hydrogel system (DECHS), which was subcutaneously injected into the hypodermis of nude mice. The PRP-loaded DECHS generated significantly more HFs (~ 35 per site) and novel vessels (~ 12 per site) than the other groups (p < 0.05 for each). Taken together, these results illustrate that, based on high-throughput microfluidics, we obtained scalable and controllable production of ECM-mimicking IGMs and DECHS, which simulate an effective micro- and macro-environment to promote DPC bioactivity and hair regeneration, thus representing a potential new strategy for HF tissue engineering.