Self-reported health problems and obesity predict sickness absence during a 12-month follow-up: a prospective cohort study in 21 608 employees from different industries.
To study whether self-reported health problems predict sickness absence (SA) from work in employees from different industries. The results of a health risk appraisal (HRA) were combined with archival data of SA of 21 608 employees (59% female, 56% clerical). Exposure variables were self-reported health problems, labelled as 'work disability (WD) risk factors' in the HRA, presence of problems with occupational well-being and obesity. Age, socioeconomic grading and the number of SA days 12 months before the survey were treated as confounders. The outcome measure was accumulated SA days during 12-month follow-up. Data were analysed separately for males and females. A Hurdle model with negative binomial response was used to analyse zero-inflated count data of SA. The HRA results predicted the number of accumulated SA days during the 12-month follow-up, regardless of occupational group and gender. The ratio of means of SA days varied between 2.7 and 4.0 among those with 'WD risk factors' and the reference category with no findings, depending on gender and occupational group. The lower limit of the 95% CI was at the lowest 2.0. In the Hurdle model, 'WD risk factors', SA days prior to the HRA and obesity were additive predictors for SA and/or the accumulated SA days in all occupational groups. Self-reported health problems and obesity predict a higher total count of SA days in an additive fashion. These findings have implications for both management and the healthcare system in the prevention of WD.


Online course increases nutrition professionals' knowledge, skills, and self-efficacy in using an ecological approach to prevent childhood obesity.
To assess the impact of an online continuing education course on the knowledge, skills, and self-efficacy of nutrition professionals to use an ecological approach to prevent childhood obesity. Quasi-experimental design using intervention and delayed intervention comparison groups with pre/post-course assessments. Online continuing education course. Nutrition and health professionals in an online course (n = 105) and a delayed intervention comparison group (n = 37). A 6-week, facilitated online course titled, Preventing Childhood Obesity: An Ecological Approach. Changes in knowledge, skills, and self-efficacy in using an ecological approach to address childhood obesity. Paired and independent sample t tests, factor analysis, regression analysis. In contrast to a comparison group, nutrition and health professionals who participated in a 6-week online course had statistically significant increases (P < .01) in their knowledge, skills, and self-efficacy related to using an ecological approach to prevent childhood obesity. A facilitated online course can be effective at increasing the knowledge, skills, and self-efficacy of community-based nutrition and health professionals in using an ecological approach to prevent childhood obesity in their communities.


Enhancement of bioactivity on medical polymer surface using high power impulse magnetron sputtered titanium dioxide film.
This study utilizes a novel technique, high power impulse magnetron sputtering (HIPIMS), which provides a higher ionization rate and ion bombardment energy than direct current magnetron sputtering (DCMS), to deposit high osteoblast compatible titanium dioxide (TiO2) coatings with anatase (A-TiO2) and rutile (R-TiO2) phases onto the biomedical polyetheretherketone (PEEK) polymer substrates at low temperature. The adhesions of TiO2 coatings that were fabricated using HIPIMS and DCMS were compared. The in vitro biocompatibility of these coatings was confirmed. The results reveal that HIPIMS can be used to prepare crystallinic columnar A-TiO2 and R-TiO2 coatings on PEEK substrate if the ratio of oxygen to argon is properly controlled. According to a tape adhesion test, the HIPIMS-TiO2 coatings had an adhesion grade of 5B even after they were immersed in simulated body fluid (SBF) environments for 28days. Scratch tests proved that HIPIMS-TiO2 coatings undergo cohesive failure. These results demonstrate that the adhesive force between HIPIMS-TiO2 coating/PEEK is stronger than that between DCMS-TiO2 coating/PEEK. After a long period (28days) of immersion in SBF, a bone-like crystallinic hydroxyapatite layer with a corresponding Ca/P stoichiometry was formed on both HIPIMS-TiO2. The osteoblast compatibility of HIPIMS-TiO2 exceeded that of the bare PEEK substrate. It is also noticeable that the R-TiO2 performed better in vitro than the A-TiO2 due to the formation of many negatively charged hydroxyl groups (-OH(-)) groups on R-TiO2 (110) surface. In summary, the HIPIMS-TiO2 coatings satisfied the requirements for osseointegration, suggesting the possibility of using HIPIMS to modify the PEEK surface with TiO2 for spinal implants.


Determination of single-kidney glomerular filtration rate (GFR) with CT urography versus renal dynamic imaging Gates method.
To present a single-kidney CT-GFR measurement and compare it with the renal dynamic imaging Gates-GFR. Thirty-six patients with hydronephrosis referred for CT urography and 99mTc-DTPA renal dynamic imaging were prospectively included. Informed consent was obtained from all patients. The CT urography protocol included non-contrast, nephrographic, and excretory phase imaging. The total CT-GFR was calculated by dividing the CT number increments of the total urinary system between the nephrographic and excretory phase by the products of iodine concentration in the aorta and the elapsed time, then multiplied by (1- Haematocrit). The total CT-GFR was then split into single-kidney CT-GFR by a left and right kidney proportionality factor. The results were compared with single-kidney Gates-GFR by using paired t-test, correlation analysis, and Bland-Altman plots. Paired difference between single-kidney CT-GFR (45.02 ± 13.91) and single-kidney Gates-GFR (51.21 ± 14.76) was 6.19 ± 5.63 ml/min, p<0.001, demonstrating 12.1% systematic underestimation with ±11.03 ml/min (±21.5%) measurement deviation. A good correlation was revealed between both measurements (r=0.87, p<0.001). The proposed single-kidney CT-GFR correlates and agrees well with the reference standard despite a systematic underestimation, therefore it could be a one-stop-shop for evaluating urinary tract morphology and split renal function. • A new CT method can assess split renal function • Only using images from CT urography and the value of haematocrit • A one-stop-shop CT technique without additional radiation dose.


The influence of different 5-methoxyindoles on the process of protein/peptide secretion characterized by the formation of granular vesicles in the mouse pineal gland. An in vitro study.
The effects of different 5-methoxyindoles on the process of protein/peptide secretion characterized by the formation of granular vesicles (GV) have been studied in mouse pinealocytes maintained in explant culture. All 5-methoxyindoles studied clearly influence the number of granular vesicles in the pinealocytes. Comparing all present results it appeared that, in this system, melatonin was the least effective of all 5-methoxyindoles tested in stimulating secretion. 5-Methoxyindole-3-acetic acid, irrespective of the duration of the experiment and of the presence of noradrenaline, increased the number of GV. For all other 5-methoxyindoles, 5-methoxytryptamine, 5-methoxytryptophan, 5-methoxytryptophol and melatonin, it appeared that the effects depend on the duration of application and on the presence or absence of noradrenaline in the medium. Moreover, depending on the experimental conditions and the 5-methoxyindole tested, antagonistic as well as synergistic effects between 5-methoxyindoles and noradrenaline were observed. The present results, which suggest that the 5-methoxyindoles are also active in the pineal gland itself, demonstrate that, as far as the formation of granular vesicles is concerned, there exists a very complex mechanism of regulation, involving the sympathetic innervation and the 5-methoxyindoles (which are themselves under the influence of this innervation). The physiological significance of this system is discussed in relation to a proposed working hypothesis.


The effect of chromatographically pure sample of adenosine 5'-tetraphosphate on sheep and rabbit blood platelets.
1. The commercially available trisodium salt of adenosine 5'-tetraphosphate (ATetraP) (Sigma) was found to be contaminated with ATP, ADP and AMP, and therefore unsuitable for use in platelet studies. 2. The more stable barium salt of ATetraP was converted to the ammonium salt and found to be chromatographically homogeneous. This sample was tested for its influence on sheep blood platelets in citrated-rich plasma by the photometric method. 3. The ammonium salt of ATetraP (5-105 mumol.1(-1)) induced platelet aggregation which showed no tendency towards disaggregation. 4. The log dose-response lines for ATetraP and for adenosine diphosphate were parallel. On a molar basis, the tetraphosphate and only 1.5% of the aggregating activity of ADP. 5. The initial rate of aggregation induced by the tetraphosphate was inhibited by adenosine 5'-monophosphate analogues which are selective ADP-antagonists. These compounds also dispersed aggregates produced by ATetraP. 6. Platelets made refractory to ADP were also refractory to ATetraP. 7. Like ADP, ATetraP induced the change in shape of rabbit platelets and in this respect had only 3.4% the activity of ADP. 8. It is concluded that ATetraP per se can induced platelet aggregation and platelet shape change, and appears to exert its effect at the same site on the platelet surface as does ADP.


Lunasin potentiates the effect of oxaliplatin preventing outgrowth of colon cancer metastasis, binds to α5β1 integrin and suppresses FAK/ERK/NF-κB signaling.
The effect of lunasin on colon cancer metastasis was studied using three human colon cancer cell lines in vitro and a liver metastasis model of colon cancer in vivo. Lunasin bound with α5β1 integrin and internalized into the nucleus of KM12L4 human colon cancer cells. Lunasin (10 μM) inhibited the activation of focal adhesion kinase (FAK) by 28%, 39% and 60% in RKO, HCT-116 and KM12L4 human colon cancer cells, respectively. Lunasin caused an increase in the expression of the inhibitor of kappa B alpha (IκB-α), a decrease in nuclear p50 NF-κB and a reduction in the migration of cancer cells. Lunasin (4 mg/kg bw) inhibited metastasis and potentiated the effect of oxaliplatin by reducing the expression of proliferating cell nuclear antigen. Liver metastatic nodules were reduced from 28 (PBS) to 14 (lunasin, P = 0.047) while combination of lunasin and oxaliplatin to 5 (P = 0.004). The tumor burden was reduced from 0.13 (PBS) to 0.10 (lunasin, P = 0.039) to 0.04 (lunasin + oxaliplatin, P < 0.0001). Moreover, lunasin potentiated the effect of oxaliplatin in modifying expression of proteins involved in apoptosis and metastasis including Bax, Bcl-2, IKK-α and p-p65. Lunasin inhibited metastasis of human colon cancer cells by direct binding with α5β1 integrin suppressing FAK/ERK/NF-κB signaling, and potentiated the effect of oxaliplatin in preventing the outgrowth of metastasis.


[Predictive Factors and Treatment of Aseptic Pseudoarthrosis of the Humeral Shaft].
PURPOSE OF THE STUDY Aseptic pseudoarthrosis (nonunion, PSA) of the humeral shaft is one of the serious complications in the treatment of fractures in this area. In a retrospective study, the authors described different methods of treating patients with this complication using predictive factors. MATERIAL AND METHODS Based on the definition of nonunion and the criteria, 14 patients (N=14) with the mean age of 56 years (32-78), namely 8 men and 6 women were included in the study. All patients were followed up clinically and radiologically at 1, 3, 6, 9, 12 months and every 3 months until the condition stabilized. The follow-up period was 11-72 months, with the average of 22 months. According to the AO classification, the fractures were classified as: 4-A1, 1-A2, 4-A3, 1-B1, 3-B2 and 1-C2. Based on the Weber-Čech classification, 2 cases of pseudoarthrosis were hypertrophic, 4 oligotrophic and 8 atrophic. According to the Non-Union Scoring System (NUSS), 5 patients reached the score under 25 points, 3 patients received 25-50 points, 4 patients 51-75 points and two patients over 76 points. In five cases, the fracture displacement index (FDI) was above 100%. In 11 patients, PSA was in the proximal and middle third and in three patients in the distal third of the humerus. METHODS Of the whole group (N=14), the nail was used in 5 cases (36%) - in 3 cases NUSS of up to 50 points and in 2 cases over 76 points. The plate was used in 9 cases (64%). In patients with NUSS of up to 50 points - in 1 case after conservative treatment, in 1 case for augmentation of nail, in 3 cases for augmentation of Prévot nails. In 4 cases with NUSS of 51-75 points a plate was replaced. Osteosynthesis was always supplemented by Judet's decortication or resection of atrophic bone and spongioplasty by autologous graft. RESULTS Shoulder function measured by the Constant Murley score (C-M) improved from 53 to 89 points (from the range of 36-76 points to the range of 75-100 points). The function of the elbow joint also improved, namely from the Mayo Elbow Performance Score (MEPS) of 65 points to 90 points (from the range of 45-70 points to the range of 80-100 points). In 11 cases (79%) the nonunion was healed and in 3 cases (21%) the failure to heal was reported. In the group with unhealed nonunions, in one case with NUSS of 51-75 points a thermoplastic casting was used and in two patients with NUSS above 76 points a palliative surgery was performed using a 12mm interlocking intramedullary nail. DISCUSSION The results of our study focused on the treatment of aseptic nonunion of the humeral shaft confirm the conclusions arrived at by other authors. As to the osteosynthesis materials, the plate remains the gold standard and provides sufficient stability for bone healing. For unhealed treatment-resistant PSA, we offer a stable intramedullary nail instead of prosthetic replacement or amputation, which provides good stability required for limb function. CONCLUSIONS When planning the treatment of aseptic pseudoarthrosis of the humeral shaft, it is necessary to have a thorough knowledge of etiopathogenesis with all predictive factors. The NUSS classification provides good guidance in treating this complication. Key words: aseptic nonunion, NUSS classification, predictive factors.


A competency framework for colonoscopy training derived from cognitive task analysis techniques and expert review.
Colonoscopy is a difficult cognitive-perceptual-motor task. Designing an appropriate instructional program for such a task requires an understanding of the knowledge, skills and attitudes underpinning the competency required to perform the task. Cognitive task analysis techniques provide an empirical means of deriving this information. Video recording and a think-aloud protocol were conducted while 20 experienced endoscopists performed colonoscopy procedures. "Cued-recall" interviews were also carried out post-procedure with nine of the endoscopists. Analysis of the resulting transcripts employed the constant comparative coding method within a grounded theory framework. The resulting draft competency framework was modified after review during semi-structured interviews conducted with six expert endoscopists. The proposed colonoscopy competency framework consists of twenty-seven skill, knowledge and attitude components, grouped into six categories (clinical knowledge; colonoscope handling; situation awareness; heuristics and strategies; clinical reasoning; and intra- and inter-personal). The colonoscopy competency framework provides a principled basis for the design of a training program, and for the design of formative assessment to gauge progress towards attaining the knowledge, skills and attitudes underpinning the achievement of colonoscopy competence.


Molecular cloning of rhodanese gene from soil metagenome of cold desert of North-West Himalayas: sequence and structural features of the rhodanese enzyme.
Rhodanese is a multifunctional, sulfur transferase that catalyzes the detoxification of cyanide by sulphuration in a double displacement (ping pong) mechanistic reaction. In the present study, small-insert metagenomic library from soil sample collected from Ladakh (3,000-3,600 m.a.s.l) in northwestern Himalayas, India was constructed. Function-driven screening of ~8,500 colonies led to the isolation of one esterase-positive clone (clone-est) harboring 2.43 kb insert. Sequence analysis of the insert identified two ORF's, phosM encoding phosphoesterase and rodM encoding rhodanese. The 800 bp rodM gene encoded a polypeptide of 227 amino acids (RodM). The RodM showed maximum homology with the rhodanese-like protein from Cyanobacterium synechococcus species with a score identity of only 51 %. Putative 3D structure of RodM developed by homology modeling resembles to homodimeric protein of SUD sulfur transferase of Wolinella succinogenes with properly structured active-site cysteine (Cys) residue. Rhodanese has been reported from few culturable microorganisms.


Symptom clustering in older Taiwanese children with cancer.
To derive symptom clusters occurring in a large group of older pediatric patients with cancer in Taiwan and to examine whether each cluster differed based on gender, type of cancer and disease, pain, and functional status. Descriptive, correlational study. Pediatric oncology inpatient unit and outpatient clinics in Taiwan. 144 pediatric patients with cancer, aged 10-18 years. Subjects completed the Memorial Symptom Assessment Scale 10-18, the Play Performance Scale for Children, and a demographic questionnaire. Medical records provided disease and treatment data. Cluster analysis techniques were used to identify the symptoms that clustered together by demographic characteristics, as well as disease, pain, and functional status. Symptom cluster, pain status, and functional status. Five clusters were identified from the statistical analysis. The symptoms that clustered together were somewhat surprising, and some can be explained by cultural differences. Patients with pain reported statistically significant higher distress in all five clusters. Five symptom clusters are identified in older Taiwanese children with cancer. The way and possible rationale of how these symptoms clustered together is discussed. This is the first study that used a statistical procedure to derive symptom clusters experienced by pediatric oncology patients. Knowledge from this study can provide a starting point to investigate the stability of symptom clusters with different states of disease, different populations, and over various periods of time.


Therapeutic artifact: Case series.
Artifacts are pervasive in every realm of biological science, and this includes the field of medicine. Unless explicitly recognized by scientific experts, artifacts can be found and may influence research and findings in the medical domain, just as they do in other branches of biological science. Therapeutic artifacts are changes that occur in the body as a result of medical interventions. These artifacts can sometimes pose challenges in forensic investigations, as they may be misinterpreted or mistaken for signs of trauma or foul play. This case series presents three cases illustrating different types of treatment artifacts encountered in forensic medicine. These three cases highlight the importance of considering medical interventions and therapies when interpreting autopsy findings. It is crucial for forensic experts to have a comprehensive understanding of therapeutic artifacts to accurately differentiate them from genuine injuries or pathologies. By recognizing and properly interpreting these artifacts, forensic investigations can be conducted with increased accuracy and ensure that justice is served.


Pleural malakoplakia caused by Rhodococcus equi infection in a patient after stem cell transplantation.
Malakoplakia is a disease especially of the urinary tract with typical plaques most frequently observed in the urinary bladder's mucosa. In the context of immunosuppression malakoplakia can also occur in other organs. Some of these extravesical malakoplakias are associated with an infection by Rhodococcus equi, a rare human pathogen well known from veterinary medicine. Here we present the first case of a pleural malakoplakia without lung involvement caused by a proved Rhodococcus equi infection.


Review of heterogeneous photochemical reactions of NOy on aerosol - A possible daytime source of nitrous acid (HONO) in the atmosphere.
As an important precursor of hydroxyl radical, nitrous acid (HONO) plays a key role in the chemistry of the lower atmosphere. Recent atmospheric measurements and model calculations show strong enhancement for HONO formation during daytime, while they are inconsistent with the known sources in the atmosphere, suggesting that current models are lacking important sources for HONO. In this article, heterogeneous photochemical reactions of nitric acid/nitrate anion and nitrogen oxide on various aerosols were reviewed and their potential contribution to HONO formation was also discussed. It is demonstrated that HONO can be formed by photochemical reaction on surfaces with deposited HNO3, by photocatalytic reaction of NO2 on TiO2 or TiO2-containing materials, and by photochemical reaction of NO2 on soot, humic acids or other photosensitized organic surfaces. Although significant uncertainties still exist in the exact mechanisms and the yield of HONO, these additional sources might explain daytime observations in the atmosphere.


Application of multiple DNA fingerprinting techniques to study the genetic relationships among three members of the subgenus Trypanozoon (Protozoa: Trypanosomatidae).
Three different DNA fingerprinting techniques, the mobile genetic element (MGE)-PCR, simple sequence repeat (SSR)-PCR and random amplified polymorphic DNA (RAPD)-PCR, were used to define a large set of genetic markers to study genetic similarity within and among Trypanosoma brucei, Trypanosoma equiperdum and Trypanosoma evansi strains (n=18) from China, Africa and South America and to investigate their genetic relationships. Using the three fingerprinting techniques, >890 bands (ranging in size from 0.2 to 2kb) were defined for all 18 strains of Trypanosoma. Within each of the strains, 39-59 bands were defined. The similarity coefficients between strains ranged from approximately 41 to 94%, with a mean of 65%. There was more genetic similarity among strains within T. evansi (mean of approximately 79%) compared with T. equiperdum ( approximately 65%) and T. brucei ( approximately 59%). The similarity coefficient data were used to construct the dendrogram, which revealed that (irrespective of species) the majority of strains from China and South America grouped together to the exclusion of those from Africa. The exceptions were a T. brucei strain from Africa and a T. equiperdum strain of unknown origin. Hence, employing data sets generated using the three different fingerprinting methods, it was not possible to unequivocally distinguish among T. brucei, T. evansi and T. equiperdum, although there was a tendency for T. evansi strains to group together to the exclusion of T. brucei. The findings provide support for the hypothesis that T. evansi originated from a mutated form of T. equiperdum and stimulate further investigations of the genetic make-up and evolution of members of the subgenus Trypanozoon.


Abnormalities of brain function during a nonverbal theory of mind task in schizophrenia.
Theory of mind (ToM), the specific ability to attribute thoughts and feelings to oneself and others is generally impaired in schizophrenia. Previous studies demonstrated a deficit of the attribution of intentions to others among patients having formal thought disorder. During nonverbal tasks, such a function requires both the visual perception of human figures and the understanding of their intentions. These processes are considered to involve the superior temporal sulcus and the medial prefrontal cortex, respectively. Are the functional patterns of activation associated with those processes abnormal in schizophrenia? Seven schizophrenic patients on medication performed a nonverbal attribution of intentions task as well as two matched physical logic tasks, with and without human figures, while H2O15 PET-scanning was performed. Data from the patients were compared to those of eight healthy controls matched for verbal IQ and sex. The experimental design allowed dissociating the effect of the perception of human figures from that of the attribution of intentions. During attribution of intentions, significant activations in the right prefrontal cortex were detected in the control subjects. Those activations were not found in the schizophrenic group. However, in both groups, the perception of human figure elicited bilateral activation of the occipitotemporal regions and of the posterior part of the superior temporal sulcus. Schizophrenic patients performing a nonverbal attribution of intentions task have an abnormal cerebral activity.


Interaction of proteins with weak amphoteric charged membrane surfaces: effect of pH.
Weak amphoteric charged membranes were prepared by the graft copolymerization of poly(ethylene glycol) (PEG) derivatives with pendant ionizable groups onto polyethylene (PE) porous membranes. Two types of weak amphoteric charged membranes and two types of weak single charged membranes were prepared. The pH dependence of the protein (fluorescein isothiocyanate-labeled bovine serum albumin, FITC-BSA) adsorption onto the membranes was investigated by fluorescence spectroscopy. The interfacial charge properties of the membranes and protein were also characterized at different pH values by streaming potential and electrophoretic light scattering (ELS) measurements, respectively. The adsorbed amount onto each ionic PEG chain grafted membrane showed a uniform maximum value near the isoelectric point (IEP) of the protein (pH 4.1). On both sides of the IEP (pHs 3.3 and 7.2), the adsorption experiments and zeta (zeta) potential measurements were well correlated: the contribution of electrostatic interaction was dominant for the protein adsorption behavior. In the alkaline condition (pH 10.2), the adsorption experiments contradict the zeta potential measurements. It suggested that the conformational change of protein molecule influenced the adsorption behavior. Finally, these results indicated the potential of controlling the protein-ionic PEG chain interaction on the membrane surfaces by the pH adjustment of the outer solution.


Predictors of scleral rupture and the role of vitrectomy in severe blunt ocular trauma.
We reviewed retrospectively 40 eyes that had received blunt trauma and had been explored for scleral rupture. Twenty-nine eyes had scleral rupture. Of these 29, ten had ruptures seen preoperatively. Nineteen had occult ruptures. The preoperative findings predictive of scleral rupture were a visual acuity of light perception or no light perception, an intraocular pressure of less than 10 mm Hg, hyphema, and chemosis. Of the 29 ruptures, 27 involved the superior hemisphere and 25 involved the anterior hemisphere of the globe. Ten of 29 eyes (34%) with scleral rupture and eight of 11 eyes (73%) without rupture achieved a final visual acuity of 5/200 or better over an average follow-up period of 6.7 months. Factors prognostic of ambulatory vision for eyes with ruptured and intact globes included an initial visual acuity of 5/200 or better, absence of scleral rupture, and a rupture length of less than 11 mm in eyes with ruptures. The vitrectomized eyes also had a better result, suggesting that early pars plana vitrectomy is of benefit in selected rupture cases.


Inhibition of the voltage-gated potassium channel Kv1.5 by hydrogen sulfide attenuates remodeling through S-nitrosylation-mediated signaling.
The voltage-gated K+ channel plays a key role in atrial excitability, conducting the ultra-rapid rectifier K+ current (IKur) and contributing to the repolarization of the atrial action potential. In this study, we examine its regulation by hydrogen sulfide (H2S) in HL-1 cardiomyocytes and in HEK293 cells expressing human Kv1.5. Pacing induced remodeling resulted in shorting action potential duration, enhanced both Kv1.5 channel and H2S producing enzymes protein expression in HL-1 cardiomyocytes. H2S supplementation reduced these remodeling changes and restored action potential duration through inhibition of Kv1.5 channel. H2S also inhibited recombinant hKv1.5, lead to nitric oxide (NO) mediated S-nitrosylation and activated endothelial nitric oxide synthase (eNOS) by increased phosphorylation of Ser1177, prevention of NO formation precluded these effects. Regulation of Ikur by H2S has important cardiovascular implications and represents a novel and potential therapeutic target.


Pentobarbitone stimulation of protein phosphorylation in rat brain in vitro.
Effect of pentobarbitone on protein phosphorylation in rat brain in vitro was investigated. It was observed that at short term incubations pentobarbitone greatly enhanced incorporation of 32P into cerebral phosphoproteins which came to lower level after prolonged incubation. Dose response curve of the drug on protein phosphorylation was unconventional. The drug did not appreciably affect lipid peroxidation in brain homogenates. Effect of pentobarbitone on protein phosphorylation is not due to its inhibition of mitochondrial electron flow which would have given opposite results.


Coxsackie B1 virus-induced changes in cell membrane-associated functions are not responsible for altered sensitivity to bacterial invasiveness.
To analyze the possible mechanisms by which coxsackie B1 virus infection affects the invasiveness of Shigella flexneri, we have studied the influence of intracellular levels of Na+ and K+, ATPase activity, cytoplasmic membrane potential, cAMP level and cell communication through gap junctions. 3h after adsorption of viable or UV-inactivated coxsackie B1 virus the Na(+)-K+ gradient of the cell collapsed, ATPase activity decreased, the cytoplasmic membranic potential-dependent tetraphosphonium ion uptake were reduced. No changes in cAMP or intercellular cell communication were observed. S. flexneri invasiveness in HEp-2 cell pretreated with viable or UV-inactivated coxsackie B 1 virus was enhanced, but bacterial invasiveness was unchanged in K(+)-depleted HEp-2 cells, cell cultures with high intracellular Na+ content or ouabain pre-treated cells compared to control cells. We found no correlation between the enhanced bacterial invasiveness in the early phase of coxsackie B 1 virus infection in HEp-2 cell cultures and intracellular K+ depletion, high intracellular Na+ content, inhibited Na(+)-K+ ATPase activity or membranic depolarization.


Expression, characterization and mutagenesis of a novel glutamate decarboxylase from Bacillus megaterium.
To search for a novel glutamate decarboxylase (GAD) with an optimum pH towards near-neutrality in order to improve production of gamma-aminobutyric acid (GABA) in recombinant hosts. A novel glutamate decarboxylase, BmGAD, from Bacillus megaterium was overexpressed and purified. BmGAD was approximately 53 kDa by SDS-PAGE analysis. Its optimum activity was at pH 5 and 50 °C. BmGAD had a specific activity of 59 ± 5.2 U mg(-1) at pH 6, which is the highest value reported so far. The apparent Km and Vmax values of BmGAD were 8 ± 0.5 mM and 150 ± 4.7 U mg(-1), respectively. Through site-directed mutagenesis, two BmGAD mutants (E294R and H467A) showed higher Vmax values than that of wild-type, with the values of 210 ± 6.9 and 180 ± 4.1 U mg(-1) at pH 5 and 50 °C, respectively. The unusual high activity of BmGAD at pH 6 makes it an attractive GABA-producing candidate in industrial application.


Individualization of breast cancer based on histopathological features and molecular alterations.
Histopathological findings and molecular alterations well reflect the biological properties of individual primary breast carcinomas. Specifically, pT (size of the invasive component), pN (number of metastatic lymph nodes), histological or nuclear grade, lymphovascular invasion, hormone receptors, and HER2 (c-erbB-2) gene overexpresson or amplification are known to be effective markers for assessing the risk of operable primary breast carcinoma, albeit incompletely. It is expected that additional molecular markers and novel diagnostic tools will be developed in the future to facilitate a more accurate characterization of higher risk node-negative breast carcinomas.


Duplexed sandwich immunoassays on a fiber-optic microarray.
In this paper, we describe a duplexed imaging optical fiber array-based immunoassay for immunoglobulin A (IgA) and lactoferrin. To fabricate the individually addressable array, microspheres were functionalized with highly specific monoclonal antibodies. The microspheres were loaded in microwells etched into the distal face of an imaging optical fiber bundle. Two microsphere-based sandwich immunoassays were developed to simultaneously detect IgA and lactoferrin, two innate immune system proteins found in human saliva. Individual microspheres could be interrogated for the simultaneous measurement of both proteins. The working concentration range for IgA detection was between 700 pM and 100 nM, while the working concentration range for lactoferrin was between 385 pM and 10 nM. The cross-reactivity between detection antibodies and their non-specific targets was relatively low in comparison to the signal generated by the specific binding with their targets. These results suggest that the degree of multiplexing on this fiber-optic array platform can be increased beyond a duplex.


A parallel fluid-solid coupling model using LAMMPS and Palabos based on the immersed boundary method.
The study of viscous fluid flow coupled with rigid or deformable solids has many applications in biological and engineering problems, e.g., blood cell transport, drug delivery, and particulate flow. We developed a partitioned approach to solve this coupled Multiphysics problem. The fluid motion was solved by Palabos (Parallel Lattice Boltzmann Solver), while the solid displacement and deformation was simulated by LAMMPS (Large-scale Atomic/Molecular Massively Parallel Simulator). The coupling was achieved through the immersed boundary method (IBM). The code modeled both rigid and deformable solids exposed to flow. The code was validated with the Jeffery orbits of an ellipsoid particle in shear flow, red blood cell stretching test, and effective blood viscosity flowing in tubes. It demonstrated essentially linear scaling from 512 to 8192 cores for both strong and weak scaling cases. The computing time for the coupling increased with the solid fraction. An example of the fluid-solid coupling was given for flexible filaments (drug carriers) transport in a flowing blood cell suspensions, highlighting the advantages and capabilities of the developed code.


Burst generation mediated by cholinergic input in terminal nerve-gonadotrophin releasing hormone neurones of the goldfish.
Peptidergic neurones play a pivotal role in the neuromodulation of widespread areas in the nervous system. Generally, it has been accepted that the peptide release from these neurones is regulated by their firing activities. The terminal nerve (TN)-gonadotrophin releasing hormone (GnRH) neurones, which are one of the well-studied peptidergic neurones in vertebrate brains, are characterised by their spontaneous regular pacemaker activities, and GnRH has been suggested to modulate the sensory responsiveness of animals. Although many peptidergic neurones are known to exhibit burst firing activities when they release the peptides, TN-GnRH neurones show spontaneous burst firing activities only infrequently. Thus, it remains to be elucidated whether the TN-GnRH neurones show burst activities and, if so, how the mode switching between the regular pacemaking and bursting modes is regulated in these neurones. In this study, we found that only a single pulse electrical stimulation of the neuropil surrounding the TN-GnRH neurones reproducibly induces transient burst activities in TN-GnRH neurones. Our combined physiological and morphological data suggest that this phenomenon occurs following slow inhibitory postsynaptic potentials mediated by cholinergic terminals surrounding the TN-GnRH neurones. We also found that the activation of muscarinic acetylcholine receptors induces persistent opening of potassium channels, resulting in a long-lasting hyperpolarisation. This long hyperpolarisation induces sustained rebound depolarisation that has been suggested to be generated by a combination of persistent voltage-gated Na(+) channels and low-voltage-activated Ca(2+) channels. These new findings suggest a novel type of cholinergic regulation of burst activities in peptidergic neurones, which should contribute to the release of neuropeptides.


Polycyclic aromatic hydrocarbons in upstream riverine runoff of the Pearl River Delta, China: an assessment of regional input sources.
Water samples collected from upstream tributaries of the Pearl River Delta (PRD) and from locations within the PRD (South China) were analyzed for 27 polycyclic aromatic hydrocarbons (PAHs). Average concentrations (aqueous plus particulate) of total 27 PAHs (Σ(27)PAH), 16 priority PAHs designated by the United States Environmental Protection Agency (USEPA) except naphthalene (Σ(15)PAH), and the seven carcinogenic PAHs (Σ(7)PAH) classified by the USEPA were 260 ± 410, 130 ± 310, and 15 ± 12 ng/L, respectively. Riverine PAHs were predominantly generated from coal and vegetation combustion, coke production, vehicle exhausts, and petroleum residues, accounting for 28%, 25%, 22% and 21%, respectively, on average. Upstream riverine fluxes of Σ(27)PAH and Σ(15)PAH amounted to 38.9 and 12.9 tons/year, respectively. The net contributions of Σ(27)PAH and Σ(15)PAH from sources within the PRD were estimated at 21.4 and 21.0 tons/year, respectively.


The partial adrenocortical hydroxylase deficiency syndrome in infertile women.
Eighteen infertile women desiring conception were selected for additional diagnostic studies on the basis of clinical evidence of hyperandrogenism characterized by acne and/or facial hirsutism. These 18 patients underwent stimulation tests with adrenocorticotropic hormone which demonstrated the presence of partial adrenocortical hydroxylase deficiences. All patients were then placed on glucocorticoid suppression therapy. Ten patients conceived and, of these, six required no therapy other than prednisone. Four conceived using a combination of prednisone and clomiphene therapy. Of othe eight patients who did not conceive, seven had other significant infertility factors. Therefore, of the patients without other significant fertility factors, 91% conceived. This study demonstrates that partial adrenocortical enzyme defects play a significant role in some women with clinical evidence of hyperandrogenism, and, in such women, either glococorticoid therapy alone or in combination with clomiphene is indicated.


Association between locomotive syndrome and anemia among community-dwelling older adults.
To investigate the association between locomotive syndrome and anemia among community-dwelling older adults. This cross-sectional study was conducted at specific health checkup centers in Japan between 2019 and 2020. We sent a questionnaire to older adults aged ≥65 years who participated in health checkups. A total of 2507 community-dwelling older adults were included in this study (mean age = 72.3 years, 51.4% women). Locomotive syndrome was measured using a 25-question Geriatric Locomotive Function Scale with a score range of 0-100, and was defined as ≥16 points. Anemia was defined using World Health Organization criteria, as a hemoglobin concentration of <13.0 g/dL in men and <12.0 g/dL in women. Logistic regression analyses were performed to investigate the association between locomotive syndrome and anemia, adjusted for age, sex, body mass index, depression symptoms, self-reported comorbidities (cancer, rheumatoid arthritis, knee osteoarthritis, fractures, and spinal disease), hypertension, and renal function. Of all the participants, 11.6% had locomotive syndrome and 12.8% had anemia. Even after adjustment, a relationship between locomotive syndrome and anemia was observed (adjusted odds ratio = 1.9, 95% confidence interval = 1.3-2.7). Community-dwelling older adults with anemia had a significantly higher prevalence of locomotive syndrome than those without. This finding suggests that older adults with anemia are at risk of locomotive syndrome, and appropriate measures should be taken for prevention. Geriatr Gerontol Int 2023; 23: 426-429.


Heart rate variability spectrum characteristics in children with sleep apnea.
Classic spectral analysis of heart rate variability (HRV) in pediatric sleep apnea-hypopnea syndrome (SAHS) traditionally evaluates the very low frequency (VLF: 0-0.04 Hz), low frequency (LF: 0.04-0.15 Hz), and high frequency (HF: 0.15-0.40 Hz) bands. However, specific SAHS-related frequency bands have not been explored. One thousand seven hundred and thirty-eight HRV overnight recordings from two pediatric databases (0-13 years) were evaluated. The first one (981 children) served as training set to define new HRV pediatric SAHS-related frequency bands. The associated relative power (RP) were computed in the test set, the Childhood Adenotonsillectomy Trial database (CHAT, 757 children). Their relationships with polysomnographic variables and diagnostic ability were assessed. Two new specific spectral bands of pediatric SAHS within 0-0.15 Hz were related to duration of apneic events, number of awakenings, and wakefulness after sleep onset (WASO), while an adaptive individual-specific new band from HF was related to oxyhemoglobin desaturations, arousals, and WASO. Furthermore, these new spectral bands showed improved diagnostic ability than classic HRV. Novel spectral bands provide improved characterization of pediatric SAHS. These findings may pioneer a better understanding of the effects of SAHS on cardiac function and potentially serve as detection biomarkers. New specific heart rate variability (HRV) spectral bands are identified and characterized as potential biomarkers in pediatric sleep apnea. Spectral band BW1 (0.001-0.005 Hz) is related to macro sleep disruptions. Spectral band BW2 (0.028-0.074 Hz) is related to the duration of apneic events. An adaptive spectral band within the respiratory range, termed ABW3, is related to oxygen desaturations. The individual and collective diagnostic ability of these novel spectral bands outperforms classic HRV bands.


Elicitor effects on reactive oxygen species in liquid cultures of Penicillium chrysogenum.
Activity of reactive oxygen species (ROS) was investigated in liquid cultures of Penicillium chrysogenum P2 supplemented with carbohydrates. Oligosaccharides lowered the ROS activity in all samples. The greatest effect occurred when oligosaccharides were added to samples 48 h after inoculation. The ROS decrease in the presence of oligoguluronate, oligomannuronate and mannan oligosaccharides was 18%, 36% and 54%, respectively (ROS levels varied notably with culture age and type of elicitor). The polysaccharides from which the oligosaccharides were derived showed little (5-10%) overall decrease of ROS.


Disclosing Frauds in Herbal Food Supplements Labeling: A Simple LC-MS/MS Approach to Detect Alkaloids and Biogenic Amines.
Food supplements are a category of products perceived safe and therefore commonly used by different categories of consumers without any particular attention or precaution. However, health risks associated with the consumption of supplements containing undeclared substances cannot be excluded. A variety of analytical methods are used to control supplement quality composition, but usually, these procedures are complex and time-consuming. Here, we report the results of a simple and fast liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, to detect and quantify simultaneously different categories of active molecules, such as biogenic ammines and natural alkaloids that at high doses can produce negative health effect in consumers. Three categories of products intended for body weight loss, energy boosting, and erectile dysfunction treatment, purchased through e-commerce sites and from local supermarkets, were analyzed (n = 91). The caffeine, synephrine, agmatine sulfate, yohimbine, phenethylamine, and icariin were correctly separated and identified with good precision (RSD < 20%) and recovery (89-109%). The identification and quantification of the analytes in real samples highlighted that the 26% of the samples were not compliant with labeling, confirming that frauds are very common also in the natural supplements market. This LC-MS/MS method could be easily used to test natural supplements in order to check the correct labeling and to protect consumers from potential health risks and food frauds.


Cloacal exstrophy: a report of 34 cases.
A clinical review is presented on 34 patients with cloacal exstrophy who were seen between 1963 and 1986. The patients were separated into 2 main groups: classical cloacal exstrophy (type I) and variant cloacal exstrophy (type II). In the classical cases 3 surface patterns were recognized: A-hemibladders confluent cranial to the bowel, B-hemibladders lateral to the bowel and C-hemibladders confluent caudal to the bowel. Surgical reconstruction was performed in 24 patients, with a 50 per cent survival rate. However, there was marked improvement in survival from 22 per cent between 1963 and 1978 to 90 per cent between 1979 and 1986.


Chemical composition and inherent properties of offal from calves maintained under two production systems.
The aim was to evaluate the influence of organ and calf type on the chemical composition, mineral concentration, fatty acid profile and inherent properties in offal. Organ type influenced (p<0.01) all evaluated characteristics, whereas calf type to a lesser extent affected mineral content. The highest contents of K, Zn and Fe were found in liver. The heart of veal calves contained the highest contents of Mg and Mn. In addition, the most favorable fatty acid profile was also found in hearts, by reason of the lowest SFA and highest PUFA percentage and h/H ratio. Significantly higher values of PUFA percentage, PUFA/SFA ratio, and content of CLA were found in organs of suckler beef, which is associated with the beneficial effects of grazing on the fatty acid profile. Results could help to increase the consumption of calves' offal and its utilization in meat processing.


Risk of Elevated Intraocular Pressure With Difluprednate in Patients With Non-Infectious Uveitis.
To evaluate the risk factors associated with clinically important intraocular pressure (IOP) elevation with topical difluprednate treatment in patients with non-infectious uveitis. Retrospective cohort study. Fifty-four eyes of 54 patients with non-infectious uveitis treated with topical difluprednate at the current institution were included. Demographics and clinical characteristics of uveitis patients were collected. The main outcome measure was development of clinically important IOP elevation defined as IOP ≥21 mmHg and an increase of ≥10 mmHg from baseline. A clinically important IOP elevation was observed in 17 patients (31.5%). The mean time to clinically important IOP elevation was 7.4±4.8 weeks (range 3-19). Statistically significant risk factors for incident clinically important IOP elevation were being a child (adjusted hazard ratio [aHR] 7.85 [95% CI 1.48-41.56], P = .02) and concurrent use of systemic steroids (aHR 5.31 [95% CI 1.18-24.00], P = .03). Patients with concurrent systemic corticosteroids developed clinically important IOP elevation earlier than those without systemic corticosteroid (mean 5.7±3.4 [range 3-14] vs 10.4±5.7 [range 4-19] weeks, P = .05). Incident IOP ≥30 mmHg occurred in 7 patients (13.0%). All patients responded well to the cessation of difluprednate and/or use of topical antiglaucomatous agents and no eyes required glaucoma surgery. This study demonstrated that clinically important IOP elevation is common in uveitis patients with topical difluprednate treatment. Children and patients with concurrent systemic corticosteroids are at substantial risk of developing clinically important IOP elevation.


Enzymes related to catecholamine biosynthesis in Tetrahymena pyriformis. Presence of GTP cyclohydrolase I.
We first identified GTP cyclohydrolase I activity (EC 184.108.40.206) in the ciliated protozoa, Tetrahymena pyriformis. The Vmax value of the enzyme in the cellular extract of T. pyriformis was 255 pmol mg-1 protein h-1. Michaelis-Menten kinetics indicated a positive cooperative binding of GTP to the enzyme. The GTP concentration producing half-maximal velocity was 0.8 mM. By high-performance liquid chromatography (HPLC) with fluorescence detection, a major peak corresponding to D-monapterin (2-amino-4-hydroxy-6-[(1'R,2'R)-1',2',3'-trihydroxypropyl]pteridin e, D-threo-neopterin) and minor peaks of D-erythro-neopterin and L-erythro-biopterin were found to be present in the cellular extract of Tetrahymena. Thus, it is strongly suggested that Tetrahymena converts GTP into unconjugated pteridine derivatives. In this study, dopamine was detected as the major catecholamine, while neither epinephrine nor norepinephrine was identified. Indeed, this protozoa was shown to possess the activity of a dopamine synthesizing enzyme, aromatic L-amino acid decarboxylase. On the other hand, activities of tyrosine hydroxylase or tyrosinase which converts tyrosine into dopa, the substrate of aromatic L-amino acid decarboxylase, could not be detected in this protozoa. Furthermore, neither dopamine beta-hydroxylase activity nor phenylethanolamine N-methyltransferase activity could be identified by the HPLC methods.


Vicinal glycol-staining identifies secondary granules in human normal and Chédiak-Higashi neutrophils.
Neutrophil secondary granules contain large amounts of glycoprotein. We evaluated periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of these granules after alpha-amylase digestion to assess their content of vicinal glycol-containing glycoconjugates and the usefulness of this stain as a positive stain for secondary granules. Using this method, early stages of secondary granule genesis were observed prior to completion of primary granule genesis in myelocytes. Immature secondary granules appeared round to ovoid, but irregular in outline and demonstrated strong staining of the limiting membrane and matrix material. In mature granules, matrix staining was unaltered; however, membrane staining was decreased. Some immature primary granules in promyelocytes demonstrated strong PA-TCH-SP reactivity which was masked in mature primary granules of band and segmented neutrophils. The Golgi apparatus showed progressively increasing PA-TCH-SP reactivity toward its mature surface which was often convex in promyelocytes and myelocytes and concave in segmented neutrophils. The Chédiak-Higashi secondary granules were cytochemically and morphologically similar to those of normal individuals and were not statistically decreased in number when compared to controls. They were only rarely observed contacting or fusing with giant granules which had consumed all primary granules leaving an easily detected population of secondary granules. Thus the alpha-amylase-PA-TCH-SP method demonstrates a large amount of unmasked vicinal glycol-containing glycoconjugates in neutrophil secondary granules, which allows their differentiation from primary granules.


Blastic transformation after splenectomy in a patient with nonvillous splenic marginal zone lymphoma with p53 overexpression: a case report.
A 61-year-old man with no subjective symptom was admitted to our hospital for further examination of the causes of anemia (hemoglobin, 9.5 g/dL) and thrombocytopenia (platelets, 9.2 x 10(4)/microL), which had been pointed out in a medical checkup half a year previously. A bone marrow examination showed 73% lymphoid cells. Immunophenotyping of these cells were CD19+CD20+CD3-CD5-CD10-CD23-, and light chain restriction (kappa) was positive by fluorescence-activated cell sorting analysis. A computed tomography scan showed mild splenomegaly. To confirm the diagnosis histologically, we performed a splenectomy. Finally, we diagnosed the patient's disease as nonvillous splenic marginal zone lymphoma (SMZL). A month after the splenectomy, the white blood cell count was remarkably increased to 7 x 10(4)/microL with the blastic transformation of lymphoid cells. We first treated the patient with fludarabine and then with the CHOP regimen (cyclophosphamide, hydroxydaunomycin, vincristine [Oncovin], and prednisone), but the disease was so refractory that the patient died of the disease 13 months after the splenectomy. Immunohistochemical staining and a molecular examination for p53 were carried out with specimens from the splenectomy. We found overexpression of the p53 protein in lymphoid cells and a point missense mutation in codon 280 at exon 8 that changed AGA (Arg) to AGT (Ser). This case may indicate the existence of a more aggressive subset of SMZL, suggesting a reconsideration of the roles of splenectomy and p53 overexpression in the diagnostic and therapeutic approaches to patients with SMZL.


Hypoxemia during ambulatory gynecologic surgery as evaluated by the pulse oximeter.
Pulse oximetry was used to determine the incidence of intraoperative hypoxemia in 108 patients undergoing ambulatory gynecologic operation. Eleven (10%) experienced moderate desaturation (arterial oxygen saturation less than 90%), and 5 (5%) suffered severe hypoxemic episodes (arterial oxygen saturation less than 85%). Among patient risk factors--including operation, body habitus, smoking habits, history of asthma, age, and airway characteristics--an association with moderate hypoxemia was found only with nonlaparoscopic gynecologic operation, obesity, and age over 35 years, and an association with severe hypoxemia was found only with obesity and age over 35. Among operative events--including inspired oxygen concentration, position, mode of ventilation, and anesthesia phase--an association with moderate hypoxemia was found only with the lithotomy position, manual ventilation, and arousal. The cost per patient of monitoring with a pulse oximeter is about +1.35. A cost-benefit analysis reveals that a mortality rate of 1 in 40,000 among patients who actually become moderately hypoxemic would justify the cost of monitoring arterial oxygen saturation. We conclude that pulse oximetry should be part of routine anesthetic monitoring.


The genotoxicity of sodium saccharin and sodium chloride in relation to their cancer-promoting properties.
The literature indicates that sodium saccharin is non-reactive to DNA and inactive as a gene mutagen in vitro. At elevated dose levels it is capable of producing structural disturbances in eukaryotic chromosomes in vitro, and it shows intermittent activity as a very weak germ-cell and somatic-cell mutagen in vivo. Its possible mode of action in these respects is speculated on and related to its ability to promote bladder tumours in rats at elevated dose levels. A review of the toxicology of sodium chloride reveals a profile of genotoxic activities almost identical to that of sodium saccharin. It is suggested that the recorded genotoxic and cancer-promoting activities of these chemicals will only become apparent at elevated dose levels that define them as significant contributors to the biological medium (solvent) rather than as trace xenobiotic toxins (solutes). The possible activity of acid saccharin, or of its potassium, calcium and ammonium salts, as ionic genotoxins requires urgent evaluation.


Concurrent Mutations Functionally Relevant.
Although single hotspot mutations in oncogenes have been the focus of much research, the clinical relevance of oncogenes with multiple mutations-now shown to be common-has only recently come into the spotlight.


CD4/CD8 ratio as a predictor of the response to HBV vaccination in HIV-positive patients: A prospective cohort study.
Hepatitis B virus (HBV) and human immunodeficiency virus (HIV) share transmission mechanisms and thus coinfection is frequent. Active immunization against HBV is essential in HIV patients. Reports using standard and reinforced HBV vaccination schedules vary widely in seroconversion rates depending on the characteristics of the included patients. Regional data concerning HBV vaccination in HIV patients are scarce. We aim to determine the serological response to HBV vaccination using standard schedule in HIV-positive patients and to evaluate characteristics that predict seroconversion. We performed a single centre prospective study of HBV vaccination with standard schedule in HIV-positive patients. Adults with negative markers of HBV infection were included between November 2012 and December 2014. Anti-HBs titres were measured 4-8 weeks after completion of vaccination schedule. Clinical, laboratory values and HIV characteristics were analyzed to determine their association with seroconversion and adherence to the HBV vaccination schedule. The study included 245 HIV-positive patients, 68.9% were male and the mean age was 42.1 years. A total of 80.7% of the patients had undetectable HIV viral loads, 86.1% had CD4 counts >200, and 94.7% were on HAART. The response to vaccination was positive in 62% (95% CI, 56-68%) and mean anti-HBs titres of 646 IU/ml. 85.5% of the responders had anti-HBs titres >100 IU/ml. An age less than 45 years, no tobacco use and a CD4/CD8 ratio >0.4 were associated with seroconversion in multivariate analysis. The seroconversion rates were 86% in the subgroup of patients who met these criteria. A total of 97.9% of the study population completed the vaccination schedule. The CD4/CD8 ratio was the primary factor associated with positive serological conversion in the multivariate analysis. The seroconversion rates were higher in a selected group of patients who were particularly suitable for the use of the standard HBV vaccination schedule.


Intrauterine Bigatti Shaver (IBS ® ) successful placental remnants removal, after caesarean section for a cervical pregnancy with placenta accreta.
Placenta accreta located in a caesarean section scar is difficult to remove. The Intrauterine Bigatti Shaver (IBS®) has already been proven to be effective in placental remnant removal. Our case report highlights that the IBS® is also a safe method to remove placental remnants attached to a previous caesarean section scar performed for a cervical pregnancy and associated with placenta accreta.


Exploration of mRNA nanoparticles based on DOTAP through optimization of the helper lipids.
Lipid nanoparticles (LNPs) are one of the most efficient carriers for RNA packaging and delivery, and vaccines based on mRNA-LNPs have received substantial attention since the outbreak of the COVID-19 pandemic. LNPs based on 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) have been widely used in preclinical and clinical settings. A novel non-viral gene delivery system called LNP3 was previously developed, which was composed of DOTAP, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and cholesterol. One of the helper lipids in this carrier was DOPE, which belongs to phospholipids. Given that substituting DOPE with non-phospholipids as helper lipids can increase the delivery efficiency of some LNPs, this study aimed to examine whether non-phospholipids can be formulated with DOTAP as helper lipids. It was found that monoglycerides with C14:0, C16:0, C18:0, C18:1, and C18:2 mediated mRNA transfection, and the transfection efficiency varied between C18:0, C18:1, and C18:2. Furthermore, substituting of the glycerol with other moieties such as the cholesterol or the ethanolamine similarly mediated mRNA transfection. The introduction of cholesterol can further improve the transfection capacity of some DOTAP-based LNPs. One of the best-performing formulations, LNP3-MO, was used to mediate luciferase-mRNA expression in vivo, and the luminescence signal was found to be mainly enriched in the lung and spleen. In addition, the level of SARS-CoV-2 spike antibody in the serum increased after three doses of LNP3-MO mediated SARS-CoV-2 spike mRNA. Altogether, this study demonstrates that non-phospholipids are promising helper lipids that can be formulated with DOTAP to facilitate efficient delivery of mRNAs in vitro and in vivo with organ-specific targeting.


Reactogenicity and immunogenicity of tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis vaccine (Tdap) in pregnant and nonpregnant women.
Tetanus toxoid, reduced diphtheria toxoid, and acellular pertusiss (Tdap) vaccine is recommended during each pregnancy, regardless of prior receipt. Data on reactogenicity and immunogenicity, particularly after repeated Tdap, are limited. We compared local injection-site and systemic reactions and serologic response following Tdap in (1) pregnant and nonpregnant women and (2) pregnant women by self-reported prior Tdap receipt. Pregnant women (gestational age 20-34 weeks) and nonpregnant women receiving Tdap were enrolled in this observational study. Injection-site and systemic reactions were assessed for one week post-vaccination. Pertussis toxin, filamentous hemagglutinin, pertactin, fimbriae, tetanus and diphtheria specific IgG antibody titers were determined by standardized enzyme-linked immunosorbent assay at baseline and 28 days post-vaccination. Reactogenicity and serologic responses were compared by pregnancy status, and within pregnant women by self-reported prior Tdap receipt. 374 pregnant and 225 nonpregnant women were vaccinated. Severe local or systemic reactions or "any" fever were uncommon (≤3% for both groups). Moderate/severe injection-site pain was significantly higher in pregnant (17.9%) versus nonpregnant (11.1%) women, but did not prompt a healthcare visit. Proportions of other moderate/severe or any severe reactions were not significantly higher in pregnant compared to nonpregnant women. Moderate/severe (including pain) and severe reactions were not significantly higher in pregnant women receiving repeat versus first-time Tdap. Antibody titers increased from baseline to post-vaccination for all vaccine antigens in pregnant and nonpregnant women; post-vaccination titers against pertussis toxin and filamentous hemagglutinin were significantly higher in nonpregnant versus pregnant women (p < 0.01). Tdap was well-tolerated in pregnant and nonpregnant women. Pregnant women were more likely to report moderate/severe pain at the Tdap injection-site compared with nonpregnant women, but did not necessitate medical visits. Prior Tdap receipt did not increase occurrence of moderate/severe local or systemic reactions in pregnant women. Serologic responses to all vaccine antigens were robust. Clinical Trial firstname.lastname@example.com. NCT02209623. https://clinicaltrials.gov/ct2/show/NCT02209623.


Oncogenes.
We have discussed oncogenes and their protein products which act at various sites in tumor cells, with special consideration for tumors of the head and neck. In a few systems studied, application of this knowledge has led to preliminary therapeutic interventions. While abnormal expression and function of these genes and proteins are responsible for transformation to the malignant phenotype, further study of these oncogenes will also help illuminate the mechanisms of normal cellular growth and differentiation. Although cancers of the upper aerodigestive tract are heterogeneous in origin and the multi-step process of carcinogenesis is likely to vary in different tumors, the head and neck tumor spectrum is a good model for tumorigenesis and should provide valuable insight into general carcinogenesis and normal cellular growth controls.


Glutathione S-transferase Mu 2 inhibits hepatic steatosis via ASK1 suppression.
Hepatic steatosis is the main characteristic of some liver metabolism diseases. However, unclear molecular mechanism of hepatic steatosis impedes the therapy of this hepatic steatosis. Glutathione-S-transferase mu 2 (GSTM2), as a member of phase II drug metabolizing enzymes (DMEs), regulates cellular antioxidant and detoxificant. GSTM2 was highly up-regulated in hepatic steatosis tissues and high-fat diet (HFD) fed mice. Loss-of-function GSTM2 mouse model demonstrated that GSTM2 protected mice from excess fat accumulation. Mechanistically, GSTM2 interacted with ASK1 and suppressed its phosphorylation and the activation of subsequent downstream p38-JNK signalling. Moreover, GSTM2 overexpression in the liver effectively ameliorated hepatic lipid accumulation. Therefore, we identified GSTM2 as an important negative regulator in progression of hepatic steatosis via both its detoxification/antioxidant and inhibition of ASK1-p38/JNK signalling. This study showed potential therapeutic function of the DME in progression of hepatic steatosis.


Lymphatic drainage of the external ear.
Lymphoscintigraphy reveals inconsistencies in our knowledge of the lymphatic anatomy of the external ear. Fifteen external ears from 9 unembalmed human cadavers were studied. Six percent hydrogen peroxide was used to find the lymphatic vessels using a surgical microscope. They were injected with a radio-opaque mixture, dissected, photographed, and radiographed to demonstrate lymphatic vessels in the tissue. Final results were transferred to the computer for analysis. Four groups of lymph collecting vessels were found. The anterior branch, in all specimens, drained directly or indirectly (having merged with a vessel descending from the scalp) into the preauricular lymph nodes. The superior, middle, and inferior (lobule) branches drained to their multiple first tier lymph nodes. An accurate lymphatic map of the external ear is described to upgrade our anatomic knowledge. It will be of benefit for the clinical management of malignancies in this region.


Perspective-Taking in Referential Communication: Does Stimulated Attention to Addressees' Perspective Influence Speakers' Reference Production?
In two experiments, we investigated whether speakers' referential communication benefits from an explicit focus on addressees' perspective. Dyads took part in a referential communication game and were allocated to one of three experimental settings. Each of these settings elicited a different perspective mind-set (baseline, self-focus, other-focus). In the two perspective settings, speakers were explicitly instructed to regard their addressees' (other-focus) or their own (self-focus) perspective before construing their referential message. Results evidenced speakers' egocentricity bias. Even though speakers were explicitly aware of addressees' informational need, speakers still referred to information not known to their addressee. Speakers' self-reported perspective-taking behavior correlated with their actual reference behavior. Those who reported to have regarded addressees' perspective were also less likely to have leaked information about their own knowledge and attentional state. Findings are discussed in light of speakers' egocentricity bias and the role of speaker-addressee collaboration in language production.


Pirmenol in the termination of paroxysmal supraventricular tachycardia.
The effects of pirmenol in terminating paroxysmal supraventricular tachycardia were studied in 25 patients. Pirmenol was administered as 1 or 2 injections of 50 mg to 17 patients during a spontaneous attack, or as a 50-mg bolus followed by steady infusion of 2.5 mg/min in 8 patients during a tachycardia that was induced electrophysiologically. Termination was successful in 11 of 17 patients who had a spontaneous attack and in 3 of 8 patients who had induced tachycardia. Pirmenol was effective in 3 of 5 patients with atrioventricular nodal reentrant mechanism, but in none of 3 patients with a reentrant tachycardia with a retrogradely conducting atrioventricular bypass tract. Conversion to sinus rhythm was achieved in 14 of 25 patients (56%). No hemodynamic adverse effects occurred. Pirmenol increased the atrial effective refractory period, but had little effect on conduction in the atrioventricular node and His-Purkinje system. Reentry was abolished through a block in the retrograde part of the dual atrioventricular nodal pathway, which is typical of class I antiarrhythmic agents.


Effect of isoprenaline, carbachol, and Cs+ on Na+ activity and pacemaker potential in rabbit SA node cells.
Effects of isoprenaline, carbachol, and Cs+ on intracellular Na+ activity (a(i)Na) and spontaneous action potentials were studied in multicellular and single cell preparations isolated from rabbit sinoatrial (SA) nodes. a(i)Na was measured with double-barreled Na+-selective microelectrodes and the fluorescent Na+-indicator sodium-binding benzofuran isophthalate (SBFI). In spontaneously beating cells, aiNa measured with Na+-selective microelectrodes and SBFI were 4.5 +/- 1.2 mM (means +/- SD, n = 21) in multicellular preparations and 4.0 +/- 1.1 mM (n = 16) in single cells, respectively. Measurements of a(i)Na with microelectrodes showed that isoprenaline increased a(i)Na from 4.7 +/- 1.2 to 5.5 +/- 1.6 mM (n = 16, P < 0.01) and shortened the action potential cycle length (ACL) from 338 +/- 46 to 269 +/- 35 ms (n = 16, P < 0.01). However, increasing the action potential rate by pacing produced a much smaller increase in a(i)Na. Changes in a(i)Na and ACL produced by isoprenaline were blocked by Cs+. The selective hyperpolarization-activated inward current (If) blocker ZD-7288 decreased a(i)Na from 5.2 +/- 1.0 to 4.6 +/- 1.3 mM (n = 4, P < 0.01) and prolonged ACL from 394 +/- 20 to 553 +/- 68 ms (n = 4, P < 0.01). The If blocker substantially inhibited the increase in a(i)Na produced by isoprenaline. Carbachol and Cs+ decreased aiNa from 4.6 +/- 1.4 to 3.9 +/- 1.2 mM (n = 15, P < 0.01) and from 4.9 +/- 1.0 to 3.9 +/- 1.3 mM (n = 18, P < 0.01), respectively. In addition, carbachol and Cs+ prolonged ACL from 345 +/- 44 to 587 +/- 100 ms (n = 15, P < 0.01) and from 353 +/- 30 to 464 +/- 87 ms (n = 18, P < 0.01), respectively. However, carbachol and Cs+ almost did not change a(i)Na when SA node cells became quiescent in a 25.4 mM extracellular K+ concentration. The results suggest that isoprenaline, ZD-7288, carbachol, or Cs+ might have changed a(i)Na and action potential rate by possibly stimulating or inhibiting If carried by Na+. Measurements of a(i)Na with SBFI showed that isoprenaline, carbachol, and Cs+ produced a(i)Na changes that were similar to those measured with the microelectrodes.


Update in intensive care medicine: studies that challenged our practice in the last 5 years.
During the last 5 years, new randomized trials in critically ill patients have challenged a number of traditional treatment strategies in intensive care. The authors review eight studies that helped change their medical practices.


Immunoinformatics-based multi-epitope vaccine design for the re-emerging monkeypox virus.
On May 7, 2022, WHO reported a new monkeypox case. By May 2023 over 80,000 cases had been reported worldwide outside previously endemic nations. (This primarily affected the men who have sex with men (MSM) community in rich nations). The present research aims to develop a multi-epitope vaccine for the monkeypox virus (MPXV) using structural and cell surface proteins. The first part of the research involved retrieving protein sequences. The Immune Epitope Database (IEDB) was then used to analyze the B and T lymphocyte epitopes. After analyzing the sensitizing properties, toxicity, antigenicity, and molecular binding, appropriate linkers were utilizedto connect selected epitopes to adjuvants, and the structure of the vaccine was formulated. Algorithms from the field of immunoinformatics predicted the secondary and tertiary structures of vaccines. The physical, chemical, and structural properties were refined and validated to achieve maximum stability. Molecular docking and molecular dynamic simulations were subsequently employed to assess the vaccine's efficacy. Afterward, the ability of the vaccine to interact with toll-like receptors 3 and 4 (TLR3 and TLR4) was evaluated. Finally, the optimized sequence was then introduced into the Escherichia coli (E. coli) PET30A + vector. An immunoinformatics evaluation suggested that such a vaccine might be safe revealed that this vaccine is safe, hydrophilic, temperature- and condition-stable, and can stimulate innate immunity by binding to TLR3 and TLR4. Our findings suggest that the first step in MPXV pathogenesis is structural and cell surface epitopes. In this study, the most effective and promising epitopes were selected and designed throughprecision servers. Furthermore,through the utilization of multi-epitope structures and a combination of two established adjuvants, this research has the potential to be a landmarkin developing an antiviralvaccine against MPXV. However, additional in vitro and in vivo tests are required to confirm these results.


Site directed mutagenesis of StSUT1 reveals target amino acids of regulation and stability.
Plant sucrose transporters (SUTs) are functional as sucrose-proton-cotransporters with an optimal transport activity in the acidic pH range. Recently, the pH optimum of the Solanum tuberosum sucrose transporter StSUT1 was experimentally determined to range at an unexpectedly low pH of 3 or even below. Various research groups have confirmed these surprising findings independently and in different organisms. Here we provide further experimental evidence for a pH optimum at physiological extrema. Site directed mutagenesis provides information about functional amino acids, which are highly conserved and responsible for this extraordinary increase in transport capacity under extreme pH conditions. Redox-dependent dimerization of the StSUT1 protein was described earlier. Here the ability of StSUT1 to form homodimers was demonstrated by heterologous expression in Lactococcus lactis and Xenopus leavis using Western blots, and in plants by bimolecular fluorescence complementation. Mutagenesis of highly conserved cysteine residues revealed their importance in protein stability. The accessibility of regulatory amino acid residues in the light of StSUT1's compartmentalization in membrane microdomains is discussed.


Crossed and uncrossed olivocochlear pathways exacerbate temporary shifts in hearing sensitivity after narrow band sound trauma in normal ears of animals with unilateral hearing impairment.
Olivocochlear (OC) pathways have been shown to reduce the temporary threshold shifts (TTSs) caused by traumatic sounds. More recently they have been shown to exacerbate TTSs under certain conditions. One condition is the normal-hearing ear of animals with a chronic unilateral hearing loss. Testing with pure tone trauma showed that then (a) the normal-hearing ear had a lower-than-normal 'intrinsic' susceptibility to intense tones, (b) binaural trauma exacerbated TTSs in the normal-hearing ear through the activity of uncrossed OC (UOC) pathways, and (c) there was no effect on TTSs of the crossed OC (COC) pathway to the normal-hearing ear. The present study is an examination in such animals of effects with noise band trauma. The effects here confirm the previous finding that under such conditions the normal-hearing ear has a lower-than-normal susceptibility to loud sound, and binaural loud sounds exacerbate TTSs in the normal-hearing ear. They extend the previous study by demonstrating that with this traumatic sound, both COC and UOC pathways exacerbate TTSs. These effects contrast against the effects seen in animals with bilaterally normal hearing for the same noise band. Given the commonality of unilateral hearing losses in the normal human population, these data have implications for the functional effects of the OC pathways on loud sound-induced hearing damage.


In vitro - in vivo correlation of intranasal drug deposition.
In vitro - in vivo correlation (IVIVC) allows prediction of in vivo drug deposition from a nasally inhaled drug based on in vitro drug measurements. In vitro measurements include physical particle characterization and, more recently, deposition studies using anatomical models. Currently, there is a lack of IVIVC for deposition measurements in anatomical models, especially for deposition patterns in various nasal cavity regions. Therefore, improvement of in vitro and in vivo measurement methods and knowledge about nasal deposition mechanisms should help IVIVC in the future.


Mauling of the "Celtic Tiger": clinical characteristics and outcome of first-episode depression secondary to the economic recession in Ireland.
There is a dearth of studies describing clinical characteristics and outcome of patients who present with mood disorders related to economic recession. To describe a cohort of patients admitted with first-episode depression related to the Irish economic recession and compare this cohort with all other first-episode depressives admitted during the same time period (2009-2010). A cohort of 137 patients admitted with first-episode depression to an independent university teaching hospital was prospectively identified and followed up from admission over 2 years (mean follow-up 430 days, s.d. 176 days). The cohort was divided into "Celtic Tiger" (patients with first-episode depression secondary to the economic recession) and non-Celtic Tiger control patients (other first-episode depressed patients). Both groups were compared in terms of clinical characteristics at baseline and outcome over follow-up. The number of admissions due to first depressive episodes were higher in recession years 2009/10 than in pre-recession years 2008/9. Celtic Tiger patients were predominantly male and more severely depressed with more marked suicidal ideation (χ(2), p<0.001) than control patients. They were more likely to recover (χ(2), p=0.013), less likely to recur (χ(2), p<0.001) and had faster time to recovery (log rank, p<0.001) and slower time to full recurrence (log rank, p=0.001). The Celtic Tiger patients spent more time asymptomatic and less time at full and subthreshold depression levels over follow-up. Study setting of centre specializing in affective disorders treatment, retrospective nature of follow-up after initial prospective interview and lack of patient follow-up interview. The study describes a subgroup of patients with severe depression associated with economic recession with likely high suicide risk but very favourable outcome.


Cushing's syndrome: a review of diagnostic tests.
This review presents an analysis and interpretation of the published experimental data that form the basis for laboratory tests commonly used for screening, definitive diagnosis, and differential diagnosis in Cushing's syndrome. The single-dose overnight dexamethasone suppression test is excellent for screening outpatients since this test has a very low incidence of false-negative results (1.9% of 154 patients with Cushing's syndrome). The definitive diagnosis of Cushing's syndrome is best established by combining basal state measurements of the daily urine-free cortisol excretion and late evening plasma cortisol levels with the 2-mg low-dose dexamethasone suppression test. The etiology of Cushing's syndrome is best determined by combining measurements of basal state plasma adrenocorticotropin (ACTH) levels with the 8-mg high-dose dexamethasone suppression test. Under certain conditions, the basal state daily urine excretion of 17-hydroxycorticosteroids and 17-ketogenic steroids, the insulin tolerance test, and the metyrapone test may be useful in the definitive or differential diagnosis of Cushing's syndrome.


Well-defined reducible cationic nanogels based on functionalized low-molecular-weight PGMA for effective pDNA and siRNA delivery.
Nucleic acid-based gene therapy is a promising treatment option to cure numerous intractable diseases. For non-viral gene carriers, low-molecular-weight polymeric vectors generally demonstrate poor transfection performance, but benefit their final removals from the body. Recently, it was reported that aminated poly(glycidyl methacrylate) (PGMA) is one potential gene vector. Based on ethylenediamine (ED)-functionalized low-molecular-weight PGMA (denoted by PGED), a flexible strategy was herein proposed to design new well-defined reducible cationic nanogels (denoted by PGED-NGs) with friendly crosslinking reagents for highly efficient nucleic acid delivery. α-Lipoic acid (LA), one natural antioxidant in human body, was readily introduced into ED-functionalized PGMA and crosslinked to produce cationic PGED-NGs with plentiful reducible lipoyl groups. PGED-NGs could effectively complex plasmid DNA (pDNA) and short interfering RNA (siRNA). Compared with pristine PGED, PGED-NGs exhibited much better performance of pDNA transfection. PGED-NGs also could efficiently transport MALAT1 siRNA (siR-M) into hepatoma cells and significantly suppressed the cancer cell proliferation and migration. The present work indicated that reducible cationic nanogels involving LA crosslinking reagents are one kind of competitive candidates for high-performance nucleic acid delivery systems. Recently, the design of new types of high-performance nanoparticles is of great significance in delivering therapeutics. Nucleic acid-based therapy is a promising treatment option to cure numerous intractable diseases. A facile and straightforward strategy to fabricate safe nucleic acid delivery nanovectors is highly desirable. In this work, based on ethylenediamine-functionalized low-molecular-weight poly(glycidyl methacrylate), a flexible strategy was proposed to design new well-defined reducible cationic nanogels (denoted by PGED-NGs) with α-Lipoic acid, one friendly crosslinking reagent, for highly efficient nucleic acid delivery. Such PGED-NGs possess plentiful reducible lipoyl groups, effectively encapsulated pDNA and siRNA and exhibited excellent abilities of nucleic acid delivery. The present work indicated that reducible cationic nanogels involving α-lipoic acid crosslinking reagents are one kind of competitive candidates for high-performance nucleic acid delivery systems.


Using cells encapsulated in agarose microbeads to analyse nuclear structure and functions.
It is now generally agreed that the nuclei of higher eukaryotes, and particularly of mammalian cells, are highly structured and that different aspects of this structure contribute to the regulation of function (1, 2). Despite the general consensus, the key mechanisms that link nuclear structure and function have proved elusive. A major reason for this is a lack of techniques that allow nuclei to be manipulated in a way that preserves the complex architectural features that are present in vivo. Historically, significant progress in understanding the makeup of nuclei from mammalian cells has been made using cells that are permeabilised in a physiological buffer after being encapsulated in agarose microbeads. By using such beads, cells are protected from shear forces that otherwise can degrade crucial elements of the architecture that it is essential to preserve.


Initiating fibro-proliferation through interfacial interactions of myoglobin colloids with collagen in solution.
This work examines fibro-proliferation through interaction of myoglobin (Mb), a globular protein with collagen, an extracellular matrix fibrous protein. Designed colloids of Mb at pH 4.5 and 7.5 have been mixed with collagen solution at pH 7.5 and 4.5 in different concentrations altering their surface charges. For the Mb colloids, 100-200nm sizes have been measured from Transmission electron micrographs and zeta sizer. CD spectra shows a shift to beta sheet like structure for the protein in the colloids. Interaction at Mb/Collagen interface studied using Dilational rheology, Quartz crystal microbalance with dissipation and Differential Scanning calorimetry show that the perturbation is not only by the charge compensation arising from the difference in pH of the colloids and collagen, but also by the organized assembly of collagen at that particular pH. Results demonstrate that positive Mb colloids at pH 4.5, having more% of entrained water stabilize the collagen fibrils (pH 7.5) around them. Ensuing dehydration leads to effective cross-linking and inherently anisotropic growth of fibrils/fibres of collagen. In the case of Mb colloids at pH 7.5, the fibril formation seems to supersede the clustering of Mb suggesting that the fibro-proliferation is both pH and hydrophilic-hydrophobic balance dependent at the interface.


Bacteria as a new model system for aging studies: investigations using light microscopy.
Aging-the decline in an individual's condition over time-is at the center of an active research field in medicine and biology. Some very basic questions have, however, remained unresolved, the most fundamental being: do all organisms age? Or are there organisms that would continue to live forever if not killed by external forces? For a long time it was believed that aging only affected organisms such as animals, plants, and fungi. Bacteria, in contrast, were assumed to be potentially immortal and until recently this assertion remained untested. We used phase-contrast microscopy (on an Olympus BX61) to follow individual bacterial cells over many divisions to prove that some bacteria show a distinction between an aging mother cell and a rejuvenated daughter, and that these bacteria thus age. This indicates that aging is a more fundamental property of organisms than was previously assumed. Bacteria can now be used as very simple model system for investigating why and how organisms age.


Pancreatic beta cell line MIN6 exhibits characteristics of glucose metabolism and glucose-stimulated insulin secretion similar to those of normal islets.
Glucose-stimulated insulin secretion, glucose transport, glucose phosphorylation and glucose utilization have been characterized in the insulinoma cell line MIN6, which is derived from a transgenic mouse expressing the large T-antigen of SV40 in pancreatic beta cells. Glucose-stimulated insulin secretion occurred progressively from 5 mmol/l glucose, reached the maximal level approximately seven-fold above the basal level at 25 mmol/l, and remained at this level up to 50 mmol/l. Glucose transport was very rapid with the half-maximal uptake of 3-O-methyl-D-glucose being reached within 15 s at 22 degrees C. Glucose phosphorylating activity in the cell homogenate was due mainly to glucokinase; the Vmax value of glucokinase activity was estimated to be 255 +/- 37 nmol.h-1.mg protein-1, constituting approximately 80% of total phosphorylating activity, whereas hexokinase activity constituted less than 20%. MIN6 cells exhibited mainly the high Km component of glucose utilization with a Vmax of 289 +/- 18 nmol.h-1.mg protein-1. Thus, glucose utilization quantitatively and qualitatively reflected glucose phosphorylation in MIN6 cells. In contrast, MIN7 cells, which exhibited only a small increase in insulin secretion in response to glucose, had 4.7-fold greater hexokinase activity than MIN6 cells with a comparable activity of glucokinase. These characteristics of MIN6 cells are very similar to those of isolated islets, indicating that this cell line is an appropriate model for studying the mechanism of glucose-stimulated insulin secretion in pancreatic beta cells.


Acute aortic dissection in a patient with Marfan's syndrome complicated by gestational hypertension.
To present a case of aortic dissection in a pregnant woman with Marfan's syndrome. A 34-year-old woman with Marfan's syndrome developed gestational hypertension and Type III aortic dissection during the third trimester. The diagnosis was confirmed by computed tomography and aortography. Although the dissection progressed, the woman was treated successfully. Elective caesarean section was performed. At follow-up the dissection had healed. Marfan's syndrome introduces a high risk of aortic dissection during pregnancy and the treatment chosen is crucial. As dilatation of the aorta is progressive, patients with Marfan's syndrome should be followed up closely and offered prophylactic surgery if necessary.


Clinical outcomes after endovascular thrombectomy in different triage methods.
The purpose of this study was to evaluate the effectiveness and safety of drip and ship (DS) for acute ischemic stroke (AIS) by comparing three treatment strategies: 1) patients seen at a primary stroke center, started on emergency intravenous thrombolysis and then transported to a comprehensive stroke center (drip and ship, DS); 2) patients immediately transferred to comprehensive stroke center without starting intravenous thrombolysis, for mechanical thrombectomy (non-drip and ship, non-DS); and 3) patients admitted directly to the comprehensive stroke center for assessment and subsequent bridging thrombolysis (mothership, MS). We retrospectively reviewed the data of patients that underwent mechanical thrombectomy for AIS from November 2020 to May 2022 at our institution. Patients were divided into three groups: DS, non-DS, and MS. Time course, multimodal CT features and clinical results were compared among the three groups. The study included 62 patients, with 19, 18, and 25 patients in DS, non-DS, and MS groups, respectively. Baseline characteristics did not differ among the three groups. The DS group had a significantly longer median onset to groin time than the MS group (395 min vs 244 min; P < 0.001), a significantly shorter onset to primary stroke center time than the non-DS group (90 min vs 463 min; P < 0.001), and a longer primary stroke center to groin puncture time than the non-DS group (277 min vs 162 min; P = 0.002). The onset to needle time was longer in the MS group than the DS group (151.2 min vs 111.8 min; P = 0.041). The intravenous thrombolysis to puncture time was shorter in the MS group compared with DS (56 min vs 278 min; P < 0.001). No significant differences were present among groups in post-operative variables measured. DS is a safe and effective method, with no increased risk of postoperative complications or death compared to non-DS and MS methods. The study provides a reference for the selection of transport modes for AIS patients.


Mastoparan, a wasp venom, activates platelets via pertussis toxin-sensitive GTP-binding proteins.
Mastoparan induced limited release of serotonin from intact human platelets, while neither intracellular calcium ion elevation nor arachidonic acid mobilization was observed. Cytolysis induced by mastoparan was negligible in the concentration range that induced serotonin release. In digitonin-permeabilized cells, mastoparan induced Ca(++)-independent release of serotonin and Ca(++)-dependent arachidonic acid release. Both serotonin release and arachidonic acid release were reduced by pertussis toxin, suggesting that platelet activation induced by mastoparan is mediated by GTP-binding proteins.


The journey of a sandwich: computer-based laboratory experiments about the human digestive system in high school biology teaching.
Teaching high school students about the digestive system can be a challenge for a teacher when s/he wants to overcome rote learning of facts without a deeper understanding of the physiological processes inside the alimentary tract. A series of model experiments illustrating the journey of a sandwich was introduced into teaching high school biology. Using a computer equipped with a commercially available data-acquisition system and a couple of sensors, it was possible to illustrate the basic underlying physical and chemical principles of digestion to the students. Students were able to investigate, through hands-on activities, the chewing force of the jaws, importance of the mechanical breakdown of food, enzymatic activity of pepsin and amylase, antibacterial activity of hydrochloric acid, and importance of the villi for absorption. Students found the experiments interesting and helpful for understanding the digestive process. Furthermore, the results from testing indicated that the students had a deeper understanding of the physiological processes.


Preliminary use of the Leo stent in the endovascular treatment of wide-necked cerebral aneurysms.
Currently available stents for intracranial use usually are Neuroform (Boston Scientific/Target, Fremont, CA) and Leo (Balt, Montmorency, France) stents. We present the results of our initial experience in using the Leo stent to treat patients with wide-necked cerebral aneurysms. Fifteen consecutive patients with wide-necked intracranial aneurysm were treated with a combined approach that consisted of delivery of a flexible self-expending neurovascular stent through a microcatheter to cover the neck of the aneurysm and subsequent filling of the aneurym with coils through the stent interstices. We assessed the clinical history, aneurysm dimensions, and technical details of the procedures, including any difficulties with stent deployment, degree of aneurysm occlusion, and complications. During a 1-year period, 15 patients with 17 broad-necked aneurysms (n = 13; average neck length, 5.1 mm; average aneruysm size, 9 mm) were treated with the Leo stent. Fifteen stents were deployed successfully; two failed. Of the coiled aneurysms, complete or subtotal (more than 95%) occlusion was achieved in 11 aneurysms, and partial occlusion was achieved in 2 aneurysms. One patient had multiple stents placed. One patient had a ruptured aneurysm at the time of treatment. Technical problems included difficulty in deploying the stent (n = 2). Two periprocedural thromboembolic complications occurred. One patient had palsy after thrombolysis was attempted. The other patient made an excellent functional recovery after undergoing successful thrombolysis of a thrombosed internal carotid artery stent. Preliminary data demonstrated that the Leo stent is useful device for the treatment of patients with wide-necked aneurysms. In cases with tortuous cerebral vasculature, delivery and deployment may be technically challenging. Clinically significant complications are uncommon.


Knee and foot contracture occur earliest in children with cerebral palsy: a longitudinal analysis of 2,693 children.
Background and purpose - Joint contracture is a common problem among children with cerebral palsy (CP). To prevent severe contracture and its effects on adjacent joints, it is crucial to identify children with a reduced range of motion (ROM) early. We examined whether significant hip, knee, or foot contracture occurs earliest in children with CP.Patients and methods - This was a longitudinal study involving 27,230 measurements obtained for 2,693 children (59% boys, 41% girls) with CP born 1990 to 2018 and registered before 5 years of age in the Swedish surveillance program for CP. The analysis was based on 4,751 legs followed up for an average of 5.0 years. Separate Kaplan-Meier (KM) curves were drawn for each ROM to illustrate the proportions of contracture-free legs at a given time during the follow-up. Using a clustered bootstrap method and considering the child as the unit of clustering, 95% pointwise confidence intervals were generated for equally spaced time points every 2.5 years for each KM curve.Results - Contracture developed in 34% of all legs, and the median time to the first contracture was 10 years from the first examination. Contracture was most common in children with a higher Gross Motor Function Classification System (GMFCS) level. The first contracture was a flexion contracture preventing dorsiflexion in children with GMFCS level I or II and preventing knee extension in children with GMFCS level III to V.Interpretation - Early interventions to prevent knee and foot contractures in children with CP should be considered.


Impact of Visual Thinking Strategies (VTS) on the Analysis of Clinical Images: A Pre-Post Study of VTS in First-Year Medical Students.
To assess the effectiveness of Visual Thinking Strategies (VTS) in medical education curricula, a pretest-posttest experimental study design was used to evaluate the impact of participating in VTS workshops on first-year medical students. A total of forty-one intervention and sixty comparative students completed the study which included the analysis of clinical images followed by a measurement of word count, length of time analyzing images, and quality of written observations of clinical images. VTS training increased the total number of words used to describe clinical images, the time spent analyzing the images, and the number of clinically relevant observations.