Human health hazards associated with the administration of antimicrobials to slaughter animals. Part II. An assessment of the risks of resistant bacteria in pigs and pork.
Risks for the consumer regarding the acquisition of resistant bacteria and/or resistance genes via the consumption of pork are discussed. In general, Salmonella spp. and Escherichia coli that originate from animals do not easily transfer their resistance genes to the resident intestinal flora of humans. The prevalence of resistant E. coli in humans seems more associated with being a vegetarian (odds ratio (OR) 1.89) than with the consumption of meat and meat products. Other risk factors are treatment with antimicrobials (OR 2-5), becoming hospitalized (OR 5.93), or working in a health setting (OR 4.38). In the Netherlands, annually an estimated 45,000 people (0-150,000) become a carrier of resistant E. coli and/or resistance genes that ori ginate from pigs, while an estimated 345,000 persons (175,000-600,000) become a carrier of resistant E. coli and/or resistance genes that originate from hospitals, e.g. other patients. Any problems with resistant Salmonella spp. that stem from pigs are, in fact, an integral part of the total problem of food-borne salmonellosis. Sometimes there are outbreaks of a specific multi-resistant clone of S. typhimurium that causes problems in both farm animals and humans. The probability that in the next 30 years there is no or maximally one outbreak of a specific clone that originates from pig herds is estimated at about 75%. Antimicrobials used as a growth promoter can have a measurable influence on the prevalence of resistant bacteria. The likely chain of events regarding avoparcin and the selection and dissemination of resistance against vancomycin in the enterococci gives the impression that the impact of the use of antimicrobials in animals on the prevalence of resistance in humans is largely determined by whether resistance genes are, or become, located on a self-transferable transposon. Furthermore, consumer health risks of antimicrobials used in slaughter pigs are mainly determined by the selection and dissemination of bacterial resistance and much less by the toxicological properties of any residues in pork. It is also concluded that most of the problems with resistant bacteria in humans are associated with the medical use of antimicrobials, and that the impact of particularly the veterinary use of antimicrobials is limited. However, the impact of antimicrobials used as a feed additive appears to be much greater than that of antimicrobials used for strictly veterinary purposes. The use of antimicrobials as a feed additive should therefore be seriously reconsidered.


Nutrient-induced stimulation of protein synthesis in mouse skeletal muscle is limited by the mTORC1 repressor REDD1.
In skeletal muscle, the nutrient-induced stimulation of protein synthesis requires signaling through the mechanistic target of rapamycin complex 1 (mTORC1). Expression of the repressor of mTORC1 signaling, regulated in development and DNA damage 1 (REDD1), is elevated in muscle during various atrophic conditions and diminished under hypertrophic conditions. The question arises as to what extent REDD1 limits the nutrient-induced stimulation of protein synthesis. The objective was to examine the role of REDD1 in limiting the response of muscle protein synthesis and mTORC1 signaling to a nutrient stimulus. Wild type REDD1 gene (REDD1(+/+)) and disruption in the REDD1 gene (REDD1(-/-)) mice were feed deprived for 16 h and randomized to remain feed deprived or refed for 15 or 60 min. The tibialis anterior was then removed for analysis of protein synthesis and mTORC1 signaling. In feed-deprived mice, protein synthesis and mTORC1 signaling were significantly lower in REDD1(+/+) than in REDD1(-/-) mice. Thirty minutes after the start of refeeding, protein synthesis in REDD1(+/+) mice was stimulated by 28%, reaching a value similar to that observed in feed-deprived REDD1(-/-) mice, and was accompanied by increased phosphorylation of mTOR (Ser2448), p70S6K1 (Thr389), and 4E-BP1 (Ser65) by 81%, 167%, and 207%, respectively. In refed REDD1(-/-) mice, phosphorylation of mTOR (Ser2448), p70S6K1 (Thr389), and 4E-BP1 (Ser65) were significantly augmented above the values observed in refed REDD1(+/+) mice by 258%, 405%, and 401%, respectively, although protein synthesis was not coordinately increased. Seventy-five minutes after refeeding, REDD1 expression in REDD1(+/+) mice was reduced (∼15% of feed-deprived REDD1(+/+) values), and protein synthesis and mTORC1 signaling were not different between refed REDD1(+/+) mice and REDD1(-/-) mice. The results show that REDD1 expression limits protein synthesis in mouse skeletal muscle by inhibiting mTORC1 signaling during periods of feed deprivation and that a reduction in its expression is necessary for maximal stimulation of protein synthesis in response to refeeding.


Risk of Non-Melanoma Skin Cancer in Connective Tissue Disease and The Impact of Immunosuppressive Therapy.
The risk of skin cancer in connective tissue disease and the impact of immunosuppressive therapy on this risk has not been well studied. The objective of this study is to investigate the risk of non-melanoma skin cancer in patients with connective tissue disease and to assess the impact of immunosuppressive therapy on this risk. This is a retrospective case control cohort study of 8281 patients with connective tissue disease (systemic lupus erythematosus, Sjogren's disease and scleroderma) and 8281 age, race, and gender matched controls followed for a 5-year period between 2002-2012, who obtained their care from a large integrated multispecialty group practice in Northern California. The odds ratio for developing squamous cell skin cancer among patients with connective tissue disease was 1.47 (95% CI, 1.14-1.90) (P=0.003) while the odds ratio for developing all non-melanoma skin cancer was 1.26 (95% CI, 1.08-1.49) (P=0.005). Patients on immunosuppressive medication for at least one year had an OR of 1.69 (95% CI, 1.16-2.45) of developing non-melanoma skin cancer (P=0.006) when controlled for age, race, gender, type of connective tissue disease, smoking status, and health care utilization. Our study shows an increased risk of non-melanoma skin cancer among patients with connective tissue disease. We also note that patients on immunosuppressive therapy for at least one year had an increased incidence of non-melanoma skin cancer. Further studies are needed to confirm these findings. J Drugs Dermatol. 2020;19(5):  doi:10.36849/JDD.2020.4781.


[Invasive pulmonary aspergillosis: a study of 33 cases].
Invasive pulmonary aspergillosis (IPA) is an infectious complication appearing mainly in immunosuppressed patients, whose diagnosis is often difficult and lately made, and that usually bears a dismal prognosis. Patients diagnosed as having IPA from 1989 to 1994 were retrospectively analyzed. Probable IPA was diagnosed on the basis of a positive culture for Aspergillus together with a consistent radiological image. Confirmed IPA was diagnosed if there was, in addition to the former, a pathological examination showing Aspergillus hifae invading pulmonary parenchyma and/or pulmonary vessels. There were 25 men and 8 women with a mean age of 53.7 +/- 16.9 years (range: 22-86 years). IPA was confirmed in 11 cases and probable in 22. Sixty three percent of the patients had hematologic malignancy or solid cancer, whereas 30.3% did not have prior granulocytopenia or immunosuppressive therapy. The mean (SD) interval between admission and diagnosis was 40.2 (37.1) days (range: 1-180 days), and the diagnosis was made while the patient was still alive in 75% of the cases. Fifteen percent of the patients had extrapulmonary aspergillosis. The most frequent finding both on X-ray film of the chest and pulmonary computed tomography were bilateral multiple pulmonary nodules. Thirteen patients were treated with itraconazole, 6 with amphotericin B, 5 received both drugs, and 2 received fluconazole. Nineteen patients (57.6%) died and the case-fatality rate among treated patients was 46.1%. IPA presents mainly in immunosuppressed patients, but there was a not negligible proportion of patients lacking the classical risk factors. IPA is often a lately made diagnosis and in a quarter of the patients it is not made when the patient is alive. The most frequent radiological presentation are multiple bilateral nodules. The case-fatality rate of IPA is exceedingly high, even when if the patient has been adequately treated.


Antibiotic choice in UK general practice: rates and drivers of potentially inappropriate antibiotic prescribing.
To identify the rates of potentially inappropriate antibiotic choice when prescribing for common infections in UK general practices. To examine the predictors of such prescribing and the clustering effects at the practice level. The rates of potentially inappropriate antibiotic choice were estimated using 1 151 105 consultations for sinusitis, otitis media and externa, upper respiratory tract infection (URTI) and lower respiratory tract infection (LRTI) and urinary tract infection (UTI), using the Clinical Practice Research Datalink (CPRD). Multilevel logistic regression was used to identify the predictors of inappropriate prescribing and to quantify the clustering effect at practice level. The rates of potentially inappropriate prescriptions were highest for otitis externa (67.3%) and URTI (38.7%) and relatively low for otitis media (3.4%), sinusitis (2.2%), LRTI (1.5%) and UTI in adults (2.3%) and children (0.7%). Amoxicillin was the most commonly prescribed antibiotic for all respiratory tract infections, except URTI. Amoxicillin accounted for 62.3% of prescriptions for otitis externa and 34.5% of prescriptions for URTI, despite not being recommended for these conditions. A small proportion of the variation in the probability of an inappropriate choice was attributed to the clustering effect at practice level (8% for otitis externa and 23% for sinusitis). Patients with comorbidities were more likely to receive a potentially inappropriate antibiotic for URTI, LRTI and UTI in adults. Patients who received any antibiotic in the 12 months before consultation were more likely to receive a potentially inappropriate antibiotic for all conditions except otitis externa. Antibiotic prescribing did not always align with prescribing guidelines, especially for URTIs and otitis externa. Future interventions might target optimizing amoxicillin use in primary care.


Histamine and endotoxin contamination of hay and respirable hay dust.
Three distinct samples collected from a barn in which an outbreak of respiratory problems occurred were examined for possible etiologic agents. No causal relationship could be established from the results of this study; however histamine concentrations as high as 0.5 ng/mg for bulk hay (in the absence of measurable creatinine levels) along with 6138.3 endotoxin units/mg of hay were present in the samples. Both endotoxin and histamine could be recovered from respirable hay dust. The authenticity of the histamine found in the hay was evaluated with high-performance liquid chromatography and radioimmunoassay. Histamine release caused by hay extracts was evaluated with the use of leukocytes from the farmer and a referent. Histamine is known to modulate the immune system, but the role of occupational or environmental exposure to histamine in respiratory disease is unknown.


Fertility-sparing surgery for borderline ovarian tumors: oncologic safety and reproductive outcomes.
To evaluate the oncological safety and reproductive outcomes of patients with borderline ovarian tumors (BOTs) treated with fertility-sparing surgery. Patients with BOTs who underwent radical or fertility-sparing surgery between 1997 and 2009 were identified from an institutional database. The recurrence rates were compared between the 2 surgery groups. To compare the reproductive outcomes, all patients who underwent fertility-sparing surgery were interviewed by telephone. One hundred forty-three patients underwent radical surgery, and 155 patients underwent fertility-sparing surgery. After a median interval of 38 months from the initial surgery (range, 10-77 months), 19 patients had a recurrence. The recurrence rate was similar in the radical and fertility-sparing surgery groups (4.9% and 7.7%, respectively; P = 0.280). In the fertility-sparing surgery group, however, the main site of recurrence was the remaining ovary that was successfully salvaged with a second round of fertility-sparing surgery. Of 51 patients who attempted to conceive, 45 patients (88.2%) were successful and resulted in 54 term deliveries. Fertility-sparing surgery for BOTs is safe and can result in future pregnancies, suggesting that such surgery should be considered for young patients who desire preservation of fertility.


Hallucinations and delirium in the dental office following triazolam administration.
A rare and unusual case of hallucinations following triazolam administration is reported. A review of the literature suggests that hallucinations following triazolam are rare; this is the first report of such a reaction when triazolam was used for oral conscious sedation in dentistry. A discussion of dental implications follows with emphasis on complete medical history evaluation before administering oral sedatives. We conclude that the proper selection of oral sedation candidates, coupled with recognition and management of adverse events, is essential.


Two neglected biologic risk factors in bone grafting and implantology: high low-density lipoprotein cholesterol and low serum vitamin D.
Following a failure of a bone graft or an implant placement, the hypothesis of a biological abnormality is rarely considered as a possible cause. A systematic search of peer-reviewed literature for dyslipidemia or vitamin D deficiency may explain this lack of consideration. Excess low-density lipoprotein cholesterol (dyslipidemia) is responsible for a slower bone metabolism or lower dental implant osseointegration. In addition, vitamin D is a key factor for linking innate and adaptive immunity. Both of these factors are compromised under the conditions of vitamin D deficiency. Therefore, vitamin D deficiency slows implant osseointegration and increases the risk of graft infection. Vitamin D is also involved in immune function and therefore allergic reactions.


T cell responses are required for protection from clinical disease and for virus clearance in severe acute respiratory syndrome coronavirus-infected mice.
A dysregulated innate immune response and exuberant cytokine/chemokine expression are believed to be critical factors in the pathogenesis of severe acute respiratory syndrome (SARS), caused by a coronavirus (SARS-CoV). However, we recently showed that inefficient immune activation and a poor virus-specific T cell response underlie severe disease in SARS-CoV-infected mice. Here, we extend these results to show that virus-specific T cells, in the absence of activation of the innate immune response, were sufficient to significantly enhance survival and diminish clinical disease. We demonstrated that T cells are responsible for virus clearance, as intravenous adoptive transfer of SARS-CoV-immune splenocytes or in vitro-generated T cells to SCID or BALB/c mice enhanced survival and reduced virus titers in the lung. Enhancement of the number of virus-specific CD8 T cells by immunization with SARS-CoV peptide-pulsed dendritic cells also resulted in a robust T cell response, earlier virus clearance, and increased survival. These studies are the first to show that T cells play a crucial role in SARS-CoV clearance and that a suboptimal T cell response contributes to the pathological changes observed in SARS. They also provide a new approach to SARS vaccine design.


Pressure-induced switching properties of the iron(iii) spin-transition complex [FeIII(3-OMeSalEen)2]PF6.
We investigated the effect of an externally applied pressure on the iron(iii) Schiff-base compound [Fe(3-OMeSalEen)2]PF6 (H-3-OMeSalEen, condensation product of 3-methoxy-substituted salicylaldehyde and N-ethylethylenediamine), which at ambient pressure displays a thermal spin transition with a 3 K wide hysteresis loop centered at 164 K. Raman spectrometry revealed the occurrence of a complete spin-state switching process for a pressure of P1/2 = 8-9 kbar at room temperature. The evolution of lattice parameters as a function of pressure was followed by X-ray diffraction measurements on single crystals, highlighting the important microscopic aspects at the origin of the pressure-induced transition, i.e. an anisotropic response and a high compressibility of the HS molecular lattice. Variable temperature magnetic susceptibility measurements at different applied pressures revealed the smoothening of the spin transition curves and a linear increase of the transition temperatures by ca. 16.4 (1.0) K kbar-1, in good agreement with the Clausius-Clapeyron law. The non-negligible influence of the pressure transmitting oils on the intrinsic transition properties was also evidenced and attributed to mechanical interactions between the particles and the solidified matrix.


When does overuse of antibiotics become a tragedy of the commons?
Over-prescribing of antibiotics is considered to result in increased morbidity and mortality from drug-resistant organisms. A resulting common wisdom is that it would be better for society if physicians would restrain their prescription of antibiotics. In this view, self-interest and societal interest are at odds, making antibiotic use a classic "tragedy of the commons". We developed two mathematical models of transmission of antibiotic resistance, featuring de novo development of resistance and transmission of resistant organisms. We analyzed the decision to prescribe antibiotics as a mathematical game, by analyzing individual incentives and community outcomes. A conflict of interest may indeed result, though not in all cases. Increased use of antibiotics by individuals benefits society under certain circumstances, despite the amplification of drug-resistant strains or organisms. In situations where increased use of antibiotics leads to less favorable outcomes for society, antibiotics may be harmful for the individual as well. For other scenarios, where a conflict between self-interest and society exists, restricting antibody use would benefit society. Thus, a case-by-case assessment of appropriate use of antibiotics may be warranted.


Pheochromocytomas and anesthesia.
The management of patients with pheochromocytoma remains a challenge for the anesthetist despite the advent of new drugs and techniques. Safe anesthesia still depends on adequate preoperative preparation and assessment together with careful monitoring during the perioperative period.


Does team training work? Principles for health care.
Teamwork is integral to a working environment conducive to patient safety and care. Team training is one methodology designed to equip team members with the competencies necessary for optimizing teamwork. There is evidence of team training's effectiveness in highly complex and dynamic work environments, such as aviation and health care. However, most quantitative evaluations of training do not offer any insight into the actual reasons why, how, and when team training is effective. To address this gap in understanding, and to provide guidance for members of the health care community interested in implementing team training programs, this article presents both quantitative results and a specific qualitative review and content analysis of team training implemented in health care. Based on this review, we offer eight evidence-based principles for effective planning, implementation, and evaluation of team training programs specific to health care.


Metabolomics of Therapy Response in Preclinical Glioblastoma: A Multi-Slice MRSI-Based Volumetric Analysis for Noninvasive Assessment of Temozolomide Treatment.
Glioblastoma (GBM) is the most common aggressive primary brain tumor in adults, with a short survival time even after aggressive therapy. Non-invasive surrogate biomarkers of therapy response may be relevant for improving patient survival. Previous work produced such biomarkers in preclinical GBM using semi-supervised source extraction and single-slice Magnetic Resonance Spectroscopic Imaging (MRSI). Nevertheless, GBMs are heterogeneous and single-slice studies could prevent obtaining relevant information. The purpose of this work was to evaluate whether a multi-slice MRSI approach, acquiring consecutive grids across the tumor, is feasible for preclinical models and may produce additional insight into therapy response. Nosological images were analyzed pixel-by-pixel and a relative responding volume, the Tumor Responding Index (TRI), was defined to quantify response. Heterogeneous response levels were observed and treated animals were ascribed to three arbitrary predefined groups: high response (HR, n = 2), TRI = 68.2 ± 2.8%, intermediate response (IR, n = 6), TRI = 41.1 ± 4.2% and low response (LR, n = 2), TRI = 13.4 ± 14.3%, producing therapy response categorization which had not been fully registered in single-slice studies. Results agreed with the multi-slice approach being feasible and producing an inverse correlation between TRI and Ki67 immunostaining. Additionally, ca. 7-day oscillations of TRI were observed, suggesting that host immune system activation in response to treatment could contribute to the responding patterns detected.


Subfunctionalization of Parental Polyamine Oxidase (PAO) Genes in the Allopolyploid Tobacco Nicotiana tabacum (L.).
Polyamines play an important role in developmental and environmental stress responses in plants. Polyamine oxidases (PAOs) are flavin-adenine-dinucleotide-dependent enzymes associated with polyamine catabolism. In this study, 14 genes were identified in the tobacco genome that code for PAO proteins being named based on their sequence homology with Arabidopsis PAOs (AtPAO1-5): NtPAO1A-B; NtPAO2A-C, NtPAO4A-D, and NtPAO5A-E. Sequence analysis confirmed that the PAO gene family of the allopolyploid hybrid Nicotiana tabacum is not an exact combination of the PAO genes of the maternal Nicotiana sylvestris and paternal Nicotiana tomentosiformis ones. The loss of the N. sylvestris homeolog of NtPAO5E and the gain of an extra NtPAO2 copy, likely of Nicotiana othophora origin, was revealed. The latter adds to the few pieces of evidence suggesting that the paternal parent of N. tabacum was an introgressed hybrid of N. tomentosiformis and N. othophora. Gene expression analysis indicated that all 14 PAO genes kept their expression following the formation of the hybrid species. The homeologous gene pairs showed similar or opposite regulation depending on the investigated organ, applied stress, or hormone treatment. The data indicate that the expression pattern of the homeologous genes is diversifying in a process of subfunctionalization.


Sex, size, and prey caloric value affect diet specialization and consumption of an invasive prey by a native predator.
Escaping the control of natural enemies is thought to heavily influence the establishment success and impact of non-native species. Here, we examined how the profitability of alternative prey in combination with the presence of a competitor and predator aggressive behavior explain individual differences in diet specialization and the consumption of the invasive green porcelain crab Petrolisthes armatus by the native mud crab predator Panopeus herbstii. Results from bomb calorimetry estimates show that invasive P. armatus has high caloric value relative to alternative native prey. Laboratory assays indicated that specialization and consumption of invasive P. armatus was mostly exhibited by large, female P. herbstii, but the presence of a competitor and predator aggressiveness did not influence diet and the consumption of P. armatus. Thus, intrinsic factors (e.g., sex and body size) seem to explain consumption of P. armatus and dietary specialization in P. herbstii, more generally. Although there are still many predator individuals that do not consume P. armatus, the proportion of individuals that have begun to specialize on P. armatus suggests that for some, it has become more profitable relative to alternative native prey. Given the high caloric value of P. armatus, we suggest that it is likely that differences in the cost of its consumption, including attack, capture, and handling times relative to alternative prey, determine its net profitability to individual predators.


Physical modeling of ribosomes along messenger RNA: Estimating kinetic parameters from ribosome profiling experiments using a ballistic model.
Gene expression is the synthesis of proteins from the information encoded on DNA. One of the two main steps of gene expression is the translation of messenger RNA (mRNA) into polypeptide sequences of amino acids. Here, by taking into account mRNA degradation, we model the motion of ribosomes along mRNA with a ballistic model where particles advance along a filament without excluded volume interactions. Unidirectional models of transport have previously been used to fit the average density of ribosomes obtained by the experimental ribo-sequencing (Ribo-seq) technique in order to obtain the kinetic rates. The degradation rate is not, however, accounted for and experimental data from different experiments are needed to have enough parameters for the fit. Here, we propose an entirely novel experimental setup and theoretical framework consisting in splitting the mRNAs into categories depending on the number of ribosomes from one to four. We solve analytically the ballistic model for a fixed number of ribosomes per mRNA, study the different regimes of degradation, and propose a criterion for the quality of the inverse fit. The proposed method provides a high sensitivity to the mRNA degradation rate. The additional equations coming from using the monosome (single ribosome) and polysome (arbitrary number) ribo-seq profiles enable us to determine all the kinetic rates in terms of the experimentally accessible mRNA degradation rate.


The metabolic responses to high carbohydrate meals with different glycemic indices consumed during recovery from prolonged strenuous exercise.
This study investigated the metabolic responses to high glycemic index (HGI) or low glycemic index (LGI) meals consumed during recovery from prolonged exercise. Eight male, trained athletes undertook 2 trials. Following an overnight fast, subjects completed a 90-min run at 70% VO(2max). Meals were provided 30 min and 2 h following cessation of exercise. The plasma glucose responses to both meals were greater in the HGI trial compared to the LGI trial (P < 0.05). Following breakfast, there were no differences in the serum insulin concentrations between the trials; however, following lunch, concentrations were higher in the HGI trial compared to the LGI trial (P < 0.05). This suggests that the glycemic index of the carbohydrates consumed during the immediate post-exercise period might not be important as long as sufficient carbohydrate is consumed. The high insulin concentrations following a HGI meal later in the recovery period could facilitate further muscle glycogen resynthesis.


Diagnostic performance of FibroTest, SteatoTest and ActiTest in patients with NAFLD using the SAF score as histological reference.
Blood tests of liver injury are less well validated in non-alcoholic fatty liver disease (NAFLD) than in patients with chronic viral hepatitis. To improve the validation of three blood tests used in NAFLD patients, FibroTest for fibrosis staging, SteatoTest for steatosis grading and ActiTest for inflammation activity grading. We pre-included new NAFLD patients with biopsy and blood tests from a single-centre cohort (FibroFrance) and from the multicentre FLIP consortium. Contemporaneous biopsies were blindly assessed using the new steatosis, activity and fibrosis (SAF) score, which provides a reliable and reproducible diagnosis and grading/staging of the three elementary features of NAFLD (steatosis, inflammatory activity) and fibrosis with reduced interobserver variability. We used nonbinary-ROC (NonBinAUROC) as the main endpoint to prevent spectrum effect and multiple testing. A total of 600 patients with reliable tests and biopsies were included. The mean NonBinAUROCs (95% CI) of tests were all significant (P < 0.0001): 0.878 (0.864-0.892) for FibroTest and fibrosis stages, 0.846 (0.830-0.862) for ActiTest and activity grades, and 0.822 (0.804-0.840) for SteatoTest and steatosis grades. FibroTest had a higher NonBinAUROC than BARD (0.836; 0.820-0.852; P = 0.0001), FIB4 (0.845; 0.829-0.861; P = 0.007) but not significantly different than the NAFLD score (0.866; 0.850-0.882; P = 0.26). FibroTest had a significant difference in median values between adjacent stage F2 and stage F1 contrarily to BARD, FIB4 and NAFLD scores (Bonferroni test P < 0.05). In patients with NAFLD, SteatoTest, ActiTest and FibroTest are non-invasive tests that offer an alternative to biopsy, and they correlate with the simple grading/staging of the SAF scoring system across the three elementary features of NAFLD: steatosis, inflammatory activity and fibrosis.


Lumbosacral arachnoid cyst with tethered cord: A rare case report.
Arachnoid cysts are cerebrospinal fluid collections in the spine that can present with neurological symptoms or be discovered accidentally. Intradural location of such cysts especially in the lumbosacral region is relatively rare. The association of such cysts with other congenital anomalies such as tethered cord lends evidence to the developmental origin of arachnoid cysts. We report a case of lumbosacral arachnoid cyst with tethered cord in a 6-year-old male child and discuss the etiopathogenesis and management options.


Fine and gross motor ability in males with ADHD.
In the this study, both fine and gross motor ability of males with attention-deficit-hyperactivity disorder (ADHD) were compared with a group of control children. Three groups of males with the following ADHD subtypes: predominantly inattentive (ADHD-PI; n = 50), hyperactive/impulsive (ADHD-HI; n = 16), or combined (ADHD-C; n = 38) were compared with 39 control males. Mean ages for the control group were 10 years 4 months (SD 1 year 4 months, range 7 years 8 months to 12 years 11 months); for the ADHD-PI group, 10 years (SD 1 year 2 months, range 7 years 10 months to 13 years); for the ADHD-HI group, 9 years 11 months (SD 1 year 2 months), range 7 years 11 months to 12 years 6 months); and for the ADHD-C group 10 years 2 months (SD 1 year 4 months, range 8 to 13 years). The Australian Disruptive Behaviours Scale and Connors' Parent Rating Scale-Revised were used to assess ADHD symptomatology. Verbal IQ was estimated using two verbal subtests of the Wechsler Intelligence Scale for Children, and movement ability was assessed using the Movement Assessment Battery for Children (MABC) and the Purdue Pegboard test. Findings demonstrated that the children with ADHD had significantly poorer movement ability than control children. A high percentage of these children displayed movement difficulties consistent with developmental coordination disorder (DCD). In addition, the current study found that the type and degree of movement difficulty differed between subtypes. The Total Impairment score, as derived from the MABC, was less severe for the ADHD-HI group than the other two ADHD groups, but more severe than for the control group. Males with ADHD-PI and ADHD-C had significantly poorer fine motor ability (p < 0.001) than control males, whereas the ADHD-HI group did not differ significantly from any of the other groups. As children with ADHD only and the control group did not differ significantly on fine motor ability but were significantly better than children categorized with both ADHD and DCD, it was argued that poorer fine motor ability found in children with ADHD could not be attributed to deficits in attention and concentration, but rather to factors relating to their motor ability.


Epidermal growth factor induces the functional expression of dopamine receptors in the GH3 cell line.
GH3 cells are a clonal strain from a rat pituitary tumor that synthesizes and secretes both PRL and GH. The peculiarity of these cells is that they do not express receptors for dopamine; thus the hormone release is insensitive to the inhibitory effect of dopamine and D2 receptor agonists. Exposure of GH3 cells to epidermal growth factor for 4 consecutive days markedly altered the cell morphology, from a spherical appearance to an elongated flattened shape, and increased the cell size. These morphological changes were accompanied by the functional expression of D2 dopamine receptors as shown by the presence of a specific, saturable, and stereoselective high affinity binding for [3H]spiroperidol in epidermal growth factor-treated cells and by the fact that the selective D2 agonist quinpirole recovered the property to inhibit PRL secretion in the cell cultures exposed to the neurotrophic factor. The effect of EGF on the functional expression of D2 receptors was dose dependent (EC50 = 8 pM) and reversible. These data suggest that EGF elicits major effects on the expression of specific genes leading to the differentiation of GH3 cells into lactotroph-like cells endowed with dopamine D2 receptors.


The functional profile of young adults with suspected Developmental Coordination Disorder (DCD).
We assessed the non-academic and academic functioning of young adults with DCD, and investigated the emotional influences and the role of strategy use within this population. A random sample of 2379 adolescents and young adults aged 19-25 (1081 males [45.4%]; mean age=20.68, SD=3.42) was used to develop the instruments. From this sample, three study groups were identified (n=429) based on the Adolescents & Adults Coordination Questionnaire: probable DCD (n=135; 67.2% males), suspected borderline DCD (n=149; 51.4% males) and control (145; 70.5% males). Participants completed the Daily Life Functions Questionnaire (assessing non-academic and academic functioning), the Recent Emotional State Test (assessing feelings resulting from task performance), the Internal Factors Attributed to Success Questionnaire and the Problem Solving Questionnaire to assess strategy and executive strategy use. A MANOVA revealed statistically significant differences between-groups (F[7,422]=16.19; p<.001; η=.197); post hoc analyses revealed differences for all measures (except the Problem Solving Questionnaire), with the probable DCD and suspected borderline DCD groups performing worse than controls. Severity of motor deficits was correlated with all outcomes except strategy use. Logistic regression revealed that non-academic functioning was the most significant predictor of group placement (B=-1.32; p<.001); academic functioning/handwriting was the second most significant predictor (B=0.44; p=.047). Deficits in motor coordination continue into adulthood and have an effect on academic and non-academic function, as well as on the emotional state of the individual.


Mucocutaneous lymph node syndrome in a young adult.
Mucocutaneous lymph node syndrome (MLNS) is an acute febrile illness of unknown cause first described in Japan but more recently reported in the United States. The mean age in confirmed cases was 3.8 years (range, 3 months to 13 years), and the mortality is 1% to 2% when associated with coronary artery thrombosis and aneurysm. We describe the first case of MLNS in a young adult, with the additional clinical finding of associated arthritis.


Identification of a novel gene--SSK1--in human endothelial cells exposed to shear stress.
To identify transcriptionally regulated genes potentially involved in the effect of shear stress on endothelial gene expression, we performed a differential display analysis of mRNAs from human umbilical vein endothelial cell (HUVEC) exposed to laminar shear stress (12 dynes/cm2) in comparison to HUVEC maintained in static condition. We identified a cDNA fragment overexpressed by laminar shear stress. The full-length, SSK1, was 3653 long and encoded for a novel protein of 1050 amino acids. Northern blot demonstrates that SSK1 mRNA is expressed at high levels also in placenta, a weak transcript was present in heart, skeletal muscle, kidney and pancreas. Homology searches of the protein databases showed that SSK1 is related to numerous serine-threonine kinases. The highest homology was found with a very recently described gene, BUBR1, an analogue of BUB1, which is a kinase involved in the regulation of cell cycle. The most conserved residues in catalytic domains II, III, VIb, VII, VIII and IX of serine-threonine protein kinases were found in the C terminal region of SSK1 which further supports the kinase nature of the new protein. The putative serine-threonine kinase SSK1 may represent a tool by which mechanical forces regulates phosphorylation events within endothelial cells.


Modulation of UV-light-induced skin inflammation by D-alpha-tocopherol and L-ascorbic acid: a clinical study using solar simulated radiation.
In this clinical trial we studied whether oral supplementation with D-alpha-tocopherol (alpha-Toc), L-ascorbic acid (Asc), or alpha-Toc combined with Asc influenced the solar simulated radiation (SSR) induced skin inflammation in healthy volunteers. We investigated the following groups in a prospective, randomized and placebo controlled study: Group (1) alpha-Toc 2 g/day, group (2) Asc 3 g/day, group (3) alpha-Toc 2 g/day combined with Asc 3 g/day, and group (4) placebo. Before and 50 days after supplementation we analyzed alpha-Toc and Asc concentrations in keratinocytes. The dose response curve of UV erythema was determined by reflectance spectrophotometry and the minimal erythema dose (MED) by visual grading before and after supplementation. 50 days after supplementation alpha-Toc keratinocyte levels were increased in groups (1) and (3), Asc concentrations were elevated in groups (2) and (3), and the a/gamma-Toc ratio increased in groups (1) and (3). The dose response curve of UVR induced erythema showed a significant flattening and the MED increased from 103 +/- 29 mJ/cm2 (before supplementation) to 183 +/- 35 mJ/cm2 (after supplementation) in group (3), while there were no significant changes in groups (1) and (2) after vitamin supplementation. Alpha-Toc and Asc act synergistically in suppression of the sunburn reaction.


Group therapist countertransference to trauma and traumatogenic situations.
In this introduction to the special issue on "Group Therapist Countertransference to Trauma and Traumatogenic Situations," the author notes that for many therapists it is not only group member disclosures and re-enactments, but also the co-occurring, sociopolitical contexts in which they live that may involve traumatic challenges. He considers three historically evolving views of countertransference: (1) the "classical"position, (2) the "totalistic" view, and (3) the "intersubjective/relational" view. He views the therapist's intense emotionality (as opposed to clinical detachment) in response to trauma as inevitable, and considers the particular roles of "bystander," "perpetrator," and "victim" induced in the therapist by traumatic re-enactments in the group. He concludes with a synopsis of some of the key points made in each of the contributions to the special issue.


Metabolic dysfunction-associated steatotic liver disease in older adults is associated with frailty and social disadvantage.
The burden of metabolic dysfunction-associated steatotic liver disease (MASLD) is growing rapidly, as is the number of older adults globally. However, relatively few studies have been performed evaluating the prevalence and risk factors for MASLD in older adults. As such, we aimed to identify the prevalence of MASLD in older adults, as well as sociodemographic, clinical, functional and biochemical associations. The study population included older adults without a history of cardiovascular disease, dementia or independence-limiting functional impairment who had participated in the ASPirin in Reducing Events in the Elderly (ASPREE) randomised trial. MASLD was defined using the Fatty Liver Index (FLI). Associations were identified using Poisson regression with robust variance for FLI ≥ 60 vs FLI < 30. 9097 Australian participants aged ≥70 years had complete biochemical and anthropometric data to identify MASLD. The study population had a mean age of 75.1 ± 4.3 years and was 45.0% male. Almost one-third (33.0%) had prevalent MASLD, and the prevalence decreased with increasing age (adjusted RR [aRR] 0.96, 95% CI: 0.96-0.97). MASLD was also negatively associated with social advantage (aRR 0.94, 95% CI: 0.90-0.99) and exercise tolerance and was positively associated with diabetes mellitus (aRR: 1.22, 95% CI: 1.16-1.29), hypertension (aRR: 1.31, 95% CI: 1.22-1.41), male sex (aRR: 1.66, 95% CI: 1.57-1.74), pre-frailty (aRR: 1.99, 95% CI: 1.82-2.12) and frailty (aRR: 2.36, 95% CI: 2.16-2.56). MASLD and nonalcoholic fatty liver disease (NAFLD) results were 100% concordant. This study in a large cohort of relatively healthy community-dwelling older adults shows that MASLD is common, decreases with age and is associated with poorer metabolic health, social disadvantage and frailty.


[The treatment of middle-aged and elderly patients suffering from trochanteric fractures of the femur].
Treatment of 198 elderly patients with trochanteric fractures included different conservative and operative therapy methods. For the first time in Russia a new approach of extrafocus osteosynthesis was recommended for "decompensated" patients. It is proved, that for this category of the injured, active surgery tactics results in a considerable improvement of treatment outcome.


Frequency and clinical relevance of inconsistent code status documentation.
Accurate and complete documentation of hospitalized patients' code status is important to ensure that healthcare providers take appropriate action in the event of a cardiac arrest. Determine the frequency and clinical relevance of incomplete and inconsistent code status documentation. Point-prevalence study. Academic medical centers. Patients admitted to general internal medicine wards. Frequency and clinical relevance of inconsistent code status documentation across 5 documentation sources. Thirty-eight (20%; 95% confidence interval [CI], 14%-26%) of 187 patients had complete and consistent code status documentation. Another 27 (14%; 95% CI, 9%-19%) patients had no code status documentation. The remaining 122 (65%; 95% CI, 58%-72%) patients had at least 1 code status documentation inconsistency. Of these, 38 (20%; 95% CI, 14%-26%) patients had a clinically relevant code status documentation inconsistency. Multivariate logistic regression analysis demonstrated that increased age (odds ratio [OR] = 1.07 [95% CI, 1.05-1.10] for every 1-year increase in age, P < 0.001) and patients receiving comfort measures (OR = 9.39 [95% CI, 1.35-65.19], P = 0.02) were independently associated with a clinically relevant code status documentation inconsistency. Incomplete and inconsistent documentation of code status occurred frequently in hospitalized patients, especially elderly patients and patients receiving comfort measures. Having multiple, poorly integrated code status documentation sources leads to a significant number of concerning inconsistencies that create opportunities for healthcare providers to inappropriately deliver or withhold resuscitative measures that conflict with patients' expressed wishes. Institutions need to be aware of this potential documentation hazard and take steps to minimize code status documentation inconsistencies.


Study on the Salivary Microbial Alteration of Men With Head and Neck Cancer and Its Relationship With Symptoms in Southwest China.
This study explored the association between oral microbes and head and neck cancer (HNC) as well as symptoms related to patients with HNC before surgical treatment. Fifty-six patients with HNC and 64 matched healthy controls were recruited from West China hospital in Southwest China. The demographic, clinical, and symptom data were collected. Salivary samples were collected to determine the microbial characteristics using 16S rRNA gene sequencing. Patients with HNC presented increased Capnocytophaga abundances. The oral microbial markers as Capnocytophaga (area under the curve=0.81) achieved a high classification power between the HNC patients and healthy controls. Moreover, using Capnocytophaga in conjunction with symptom of voice/speech difficulty achieved an overall predicting accuracy of 92.5% comparing with using Capnocytophaga alone (79.2% accuracy) in distinguishing the HNC patients from healthy controls. Salivary microbial profiles and HNC symptoms may be potential biomarkers for HNC screening.


Regulation of nuclear transport: central role in development and transformation?
Transport of macromolecules into and out of the nucleus is generally effected by targeting signals that are recognized by specific members of the importin/exportin transport receptor family. The latter mediate passage through the nuclear envelope-embedded nuclear pore complexes (NPCs) by conferring interaction with NPC constituents, as well as with other components of the nuclear transport machinery, including the guanine nucleotide-binding protein Ran. Importantly, nuclear transport is regulated at multiple levels via a diverse range of mechanisms, such as the modulation of the accessibility and affinity of target signal recognition by importins/exportins, with phosphorylation/dephosphorylation as a major mechanism. Alteration of the level of the expression of components of the nuclear transport machinery also appears to be a key determinant of transport efficiency, having central importance in development, differentiation and transformation.


Neuron and neuroblast numbers and cytogenesis in the dentate gyrus of aged APPswe/PS1dE9 transgenic mice: Effect of long-term treatment with paroxetine.
Altered neurogenesis may influence hippocampal functions such as learning and memory in Alzheimer's disease. Selective serotonin reuptake inhibitors enhance neurogenesis and have been reported to reduce cerebral amyloidosis in both humans and transgenic mice. We have used stereology to assess the longitudinal changes in the number of doublecortin-expressing neuroblasts and number of granular neurons in the dentate gyrus of APPswe/PS1dE9 transgenic mice. Furthermore, we investigated the effect of long-term paroxetine treatment on the number of neuroblasts and granular neurons, hippocampal amyloidosis, and spontaneous alternation behaviour, a measure of spatial working memory, in transgenic mice. We observed no difference in granular neurons between transgenic and wild type mice up till 18months of age, and no differences with age in wild type mice. The number of neuroblasts and the performance in the spontaneous alternation task was reduced in aged transgenic mice. Paroxetine treatment from 9 to 18months of age reduced hippocampal amyloidosis without affecting the number of neuroblasts or granular neurons. These findings suggest that the amyloidosis affects the differentiation of neuroblasts and spatial working memory, independent of changes in total granular neurons. Furthermore, while long-term paroxetine treatment may be able to reduce hippocampal amyloidosis, it appears to have no effect on total number of granular neurons or spatial working memory.


Exploring social media technologies for novice EFL school teachers to collaborate and communicate: A case in the Czech Republic.
With an increasing number of international schools, traditional EFL teaching methods may not satisfy students' needs. This study aims to investigate perceptions of social media technologies (e.g., Web 2.0) and willingness to adopt such technologies to collaborate and communicate in multicultural classrooms among novice EFL schoolteachers in the Czech Republic. The participants were 100 novice EFL schoolteachers in Prague and the South Moravian regions of the Czech Republic. The study used a mixed research method consisting of a survey (stage 1) and a semi-structured interview (stage 2). The survey examined the participants' appraisal and concerns of using social media technologies to collaborate and to communicate as well as the level of willingness to use social media technologies. A hierarchical cluster analysis using participants' responses regarding their attitudes and behavioural tendency towards using Web 2.0 social media technologies in language classrooms identified three clusters of teachers. The teachers who were most likely to adopt social Web 2.0 technologies were those who had the highest ratings on both appraisals and concerns regarding the use of social media in language classrooms. The results from the semi-structured interviews were consistent with those from the survey. Together, the results from the two stages demonstrated that most pre-service teachers favoured using Web 2.0 technology for collaboration and communication among colleagues and stakeholders in a broader community, but they displayed contrasting levels of appraisal of and concerns towards using social media technologies. Participants believed that this might be due to their different levels of ICT proficiency, workload, and working environment. The political and practical implications in K-12 education in the Czech context are also discussed.


Neurogenetics of dopaminergic receptor supersensitivity in activation of brain reward circuitry and relapse: proposing "deprivation-amplification relapse therapy" (DART).
It is well known that after prolonged abstinence, individuals who use their drug of choice experience a powerful euphoria that often precipitates relapse. While a biological explanation for this conundrum has remained elusive, we hypothesize that this clinically observed "supersensitivity" might be tied to genetic dopaminergic polymorphisms. Another therapeutic conundrum relates to the paradoxical finding that the dopaminergic agonist bromocriptine induces stronger activation of brain reward circuitry in individuals who carry the DRD2 A1 allele compared with DRD2 A2 allele carriers. Because carriers of the A1 allele relative to the A2 allele of the DRD2 gene have significantly lower D2 receptor density, a reduced sensitivity to dopamine agonist activity would be expected in the former. Thus, it is perplexing that with low D2 density there is an increase in reward sensitivity with the dopamine D2 agonist bromocriptine. Moreover, under chronic or long-term therapy with D2 agonists, such as bromocriptine, it has been shown in vitro that there is a proliferation of D2 receptors. One explanation for this relates to the demonstration that the A1 allele of the DRD2 gene is associated with increased striatal activity of L-amino acid decarboxylase, the final step in the biosynthesis of dopamine. This appears to be a protective mechanism against low receptor density and would favor the utilization of an amino acid neurotransmitter precursor like L-tyrosine for preferential synthesis of dopamine. This seems to lead to receptor proliferation to normal levels and results in significantly better treatment compliance only in A1 carriers. We propose that low D2 receptor density and polymorphisms of the D2 gene are associated with risk for relapse of substance abuse, including alcohol dependence, heroin craving, cocaine dependence, methamphetamine abuse, nicotine sensitization, and glucose craving. With this in mind, we suggest a putative physiological mechanism that may help to explain the enhanced sensitivity following intense acute dopaminergic D2 receptor activation: "denervation supersensitivity." Rats with unilateral depletions of neostriatal dopamine display increased sensitivity to dopamine agonists estimated to be 30 to 100 x in the 6-hydroxydopamine (6-OHDA) rotational model. Given that mild striatal dopamine D2 receptor proliferation occurs (20%-40%), it is difficult to explain the extent of behavioral supersensitivity by a simple increase in receptor density. Thus, the administration of dopamine D2 agonists would target D2 sensitization and attenuate relapse, especially in D2 receptor A1 allele carriers. This hypothesized mechanism is supported by clinical trials utilizing amino acid neurotransmitter precursors, enkephalinase, and catechol-O-methyltransferase (COMT) enzyme inhibition, which have resulted in attenuated relapse rates in reward deficiency syndrome (RDS) probands. If future translational research reveals that dopamine agonist therapy reduces relapse in RDS, it would support the proposed concept, which we term "deprivation-amplification relapse therapy" (DART). This term couples the mechanism for relapse, which is "deprivation-amplification," especially in DRD2 A1 allele carriers with natural D2 agonist therapy utilizing amino acid precursors and COMT and enkepalinase inhibition therapy.


Using Fixation-Related Potentials for Inspecting Natural Interactions.
Brain-Computer Interfaces (BCI) offer unique windows into the cognitive processes underlying human-machine interaction. Identifying and analyzing the appropriate brain activity to have access to such windows is often difficult due to technical or psycho-physiological constraints. Indeed, studying interactions through this approach frequently requires adapting them to accommodate specific BCI-related paradigms which change the functioning of their interface on both the human-side and the machine-side. The combined examination of Electroencephalography and Eyetracking recordings, mainly by means of studying Fixation-Related Potentials, can help to circumvent the necessity for these adaptations by determining interaction-relevant moments during natural manipulation. In this contribution, we examine how properties contained within the bi-modal recordings can be used to assess valuable information about the interaction. Practically, three properties are studied which can be obtained solely through data obtained from analysis of the recorded biosignals. Namely, these properties consist of relative gaze metrics, being abstractions of the gaze patterns, the amplitude variations in the early brain activity potentials and the brain activity frequency band differences between fixations. Through their observation, information about three different aspects of the explored interface are obtained. Respectively, the properties provide insights about general perceived task difficulty, locate moments of higher attentional effort and discriminate between moments of exploration and moments of active interaction.


Prevalence of cystic echinococcosis in relatives of patients undergoing surgery for hepatic cystic echinococcosis in an endemic region.
Cystic echinococcosis (CE) is an endemic disease in southern Chile. The aim of this study was to ascertain the prevalence of CE among relatives of patients who underwent surgical intervention for this disease in Cautín, a province of southern Chile. Cross-sectional study. Relatives of patients who underwent surgery for hepatic echinococcosis (HE), who lived at the same address, during the period 2000-2020 were studied. A total of 288 relatives of 322 patients who underwent surgery for HE participated in a CE screening. All these relatives were interviewed and underwent abdominal ultrasonography, chest X-ray and immunodiagnostic studies (relatives who had been diagnosed with or had undergone surgery for CE were excluded). Descriptive statistics were applied. Prevalence calculation, odds ratio (OR), and their respective 95% confidence intervals (95% CI) were determined. Abdominal or thoracic CE was verified in 42 relatives of subjects operated on for HE (mean age 41±8 years; 73.8% women; 38.1% of cases had two or more cysts), all of them new and asymptomatic cases. CE was detected in the lungs, liver, peritoneum, and spleen in 16.7%; 71.4%; 7.1%; and 4.8%, respectively. The overall prevalence of EQ during the studied time period was 14,6% (17.9% and 12.3% in relatives of first and second degree respectively (OR:1.56; CI 95%: 0.81; 3.01). There is a high prevalence of CE in relatives of patients undergoing surgery by this disease in the province of Cautín, Chile.


Seawater acidification and elevated temperature affect gene expression patterns of the pearl oyster Pinctada fucata.
Oceanic uptake of anthropogenic carbon dioxide results in decrease in seawater pH and increase in temperature. In this study, we demonstrated the synergistic effects of elevated seawater temperature and declined seawater pH on gene expression patterns of aspein, calmodulin, nacrein, she-7-F10 and hsp70 in the pearl oyster Pinctada fucata. Under 'business-as-usual' scenarios, four treatments were examined: (1) ambient pH (8.10) and ambient temperature (27 °C) (control condition), (2) ambient pH and elevated temperature (+3 °C), (3) declined pH (7.70) and ambient temperature, (4) declined pH and elevated temperature. The results showed that under warming and acidic seawater conditions, expression of aspein and calmodulin showed no significant differences among different time point in condition 8.10 T. But the levels of aspein and calmodulin in conditions 8.10 T+3, 7.70 T and 7.70 T+3, and levels of nacrein, she-7-F10 in all the four treatments changed significantly. Low pH and pH × temperature interaction influenced the expression of aspein and calmodulin significantly after hours 48 and 96. Significant effects of low pH and pH × temperature interaction on the expression of nacrein were observed at hour 96. The expression level of she-7-F10 was affected significantly by pH after hours 48 and 96. The expression of hsp70 was significantly affected by temperature, pH, temperature × pH interaction at hour 6, and by temperature × pH interaction at hour 24. This study suggested that declined pH and pH × temperature interaction induced down regulation of calcification related genes, and the interaction between declined seawater pH and elevated temperature caused up regulation of hsp70 in P. facata. These results demonstrate that the declined seawater pH and elevated temperature will impact the physiological process, and potentially the adaptability of P. fucata to future warming and acidified ocean.


The human rhodopsin kinase promoter in an AAV5 vector confers rod- and cone-specific expression in the primate retina.
Adeno-associated virus (AAV) has proven an effective gene delivery vehicle for the treatment of retinal disease. Ongoing clinical trials using a serotype 2 AAV vector to express RPE65 in the retinal pigment epithelium have proven safe and effective. While many proof-of-concept studies in animal models of retinal disease have suggested that gene transfer to the neural retina will also be effective, a photoreceptor-targeting AAV vector has yet to be used in the clinic, principally because a vector that efficiently but exclusively targets all primate photoreceptors has yet to be demonstrated. Here, we evaluate a serotype 5 AAV vector containing the human rhodopsin kinase (hGRK1) promoter for its ability to target transgene expression to rod and cone photoreceptors when delivered subretinally in a nonhuman primate (NHP). In vivo fluorescent fundus imaging confirmed that AAV5-hGRK1-mediated green fluorescent protein (GFP) expression was restricted to the injection blebs of treated eyes. Optical coherence tomography (OCT) revealed a lack of gross pathology after injection. Neutralizing antibodies against AAV5 were undetectable in post-injection serum samples from subjects receiving uncomplicated subretinal injections (i.e., no hemorrhage). Immunohistochemistry of retinal sections confirmed hGRK1 was active in, and specific for, both rods and cones of NHP retina. Biodistribution studies revealed minimal spread of vector genomes to peripheral tissues. These results suggest that AAV5-hGRK1 is a safe and effective AAV serotype/promoter combination for targeting therapeutic transgene expression protein to rods and cones in a clinical setting.


Retigabine as add-on treatment of refractory epilepsy--a cost-utility study in a Swedish setting.
To calculate comparative incremental cost-effectiveness ratios (cost per quality-adjusted life year, QALY) and net marginal benefits for retigabine as add-on treatment for patients with uncontrolled focal seizures as compared to add-on lacosamide treatment and no add-on treatment, respectively. Calculations were performed using a validated decision-tree model. The study population consisted of adult patients with focal-onset epilepsy in published randomized placebo-controlled add-on trials of retigabine or lacosamide. Healthcare utilization and QALY for each treatment alternative were calculated. Probabilistic sensitivity analysis was performed using the specification of this model as a basis for Monte Carlo simulations. 2009 prices were used for all costs. Results were reported for a 2-year follow-up period. Retigabine add-on treatment was both more effective and less costly than lacosamide add-on treatment, and the cost per additional QALY for the retigabine no add-on (standard) therapy comparison was estimated at 2009€ 15,753. Using a willingness-to-pay threshold for a QALY of € 50,000, the net marginal values were estimated at 2009€ 605,874 for retigabine vs lacosamide and 2009€ 2,114,203 for retigabine vs no add-on, per 1,000 patients. The probabilistic analyses showed that the likelihood that retigabine treatment is cost-effective is at least 70%. The estimated cost per additional QALY, for the retigabine vs no add-on treatment comparison, is well within the range of newly published estimates of willingness to pay for an additional QALY. Thus, add-on retigabine treatment for people with focal-onset epilepsy with no/limited response to standard antiepileptic treatment appears to be cost-effective.


Strigea falconispalumbi (Trematoda: Strigeidae): argentophilic structures of the miracidium.
A study of the argentophilic structures of the miracidium of the strigeid digenean Strigea falconispalumbi VIBORG, 1795 was carried out. Miracidia were hatched under laboratory conditions from eggs obtained from adult S. falconispalumbi from the intestine of a naturally infected Buzzard (Buteo buteo). Miracidia were stained with 0.5%. Silver Nitrate solution using standard techniques. The epidermal plates of the miracidium are arranged in four tiers according to the formula (6 + 8 + 4 + 3) = 21. The terabratorium bears 20 papillare. Papillae are also present at the bases of the epidermal plates of the first tier, and also at the margins of the terabratorium between the epidermal plates. The two pores of the excretory ducts are positioned laterally between the third and fourth tiers of epidermal plates.


Association of circulating PLA2G7 levels with cancer cachexia and assessment of darapladib as a therapy.
Cancer cachexia (CCx) is a multifactorial wasting disorder characterized by involuntary loss of body weight that affects many cancer patients and implies a poor prognosis, reducing both tolerance to and efficiency of anticancer therapies. Actual challenges in management of CCx remain in the identification of tumour-derived and host-derived mediators involved in systemic inflammation and tissue wasting and in the discovery of biomarkers that would allow for an earlier and personalized care of cancer patients. The aim of this study was to identify new markers of CCx across different species and tumour entities. Quantitative secretome analysis was performed to identify specific factors characteristic of cachexia-inducing cancer cell lines. To establish the subsequently identified phospholipase PLA2G7 as a marker of CCx, plasma PLA2G7 activity and/or protein levels were measured in well-established mouse models of CCx and in different cohorts of weight-stable and weight-losing cancer patients with different tumour entities. Genetic PLA2G7 knock-down in tumours and pharmacological treatment using the well-studied PLA2G7 inhibitor darapladib were performed to assess its implication in the pathogenesis of CCx in C26 tumour-bearing mice. High expression and secretion of PLA2G7 were hallmarks of cachexia-inducing cancer cell lines. Circulating PLA2G7 activity was increased in different mouse models of CCx with various tumour entities and was associated with the severity of body wasting. Circulating PLA2G7 levels gradually rose during cachexia development. Genetic PLA2G7 knock-down in C26 tumours only partially reduced plasma PLA2G7 levels, suggesting that the host is also an important contributor. Chronic treatment with darapladib was not sufficient to counteract inflammation and tissue wasting despite a strong inhibition of the circulating PLA2G7 activity. Importantly, PLA2G7 levels were also increased in colorectal and pancreatic cancer patients with CCx. Overall, our data show that despite no immediate pathogenic role, at least when targeted as a single entity, PLA2G7 is a consistent marker of CCx in both mice and humans. The early increase in circulating PLA2G7 levels in pre-cachectic mice supports future prospective studies to assess its potential as biomarker for cancer patients.


Cardiac surgery in a patient with severe thrombocytopenia: how low is too low?
Platelets play a very important role in hemostasis, especially after cardiac surgery. Excessive bleeding after such surgery may lead to increased need for transfusion and its incumbent increase in post-operative morbidity and mortality. Although most cardiac surgeons will offer a surgical option to a patient with moderate thrombocytopenia (platelet count around 70 × 10 9 /L), successful cardiac surgery has not been reported in patients with significantly lower platelets counts (less than 40 × 10 9 /L). We report a case of severe thrombocytopenia (19 × 10 9 /L) where coronary artery bypass grafting was performed with minimal blood loss post-operatively, discuss the patient's management and provide insights while dealing with such patients.


Monitoring of an esterification reaction by on-line direct liquid sampling mass spectrometry and in-line mid infrared spectrometry with an attenuated total reflectance probe.
A specially designed thermal vaporiser was used with a process mass spectrometer designed for gas analysis to monitor the esterification of butan-1-ol and acetic anhydride. The reaction was conducted at two scales: in a 150 mL flask and a 1L jacketed batch reactor, with liquid delivery flow rates to the vaporiser of 0.1 and 1.0 mLmin(-1), respectively. Mass spectrometry measurements were made at selected ion masses, and classical least squares multivariate linear regression was used to produce concentration profiles for the reactants, products and catalyst. The extent of reaction was obtained from the butyl acetate profile and found to be 83% and 76% at 40°C and 20°C, respectively, at the 1L scale. Reactions in the 1L reactor were also monitored by in-line mid-infrared (MIR) spectrometry; off-line gas chromatography (GC) was used as a reference technique when building partial least squares (PLS) multivariate calibration models for prediction of butyl acetate concentrations from the MIR spectra. In validation experiments, good agreement was achieved between the concentration of butyl acetate obtained from in-line MIR spectra and off-line GC. In the initial few minutes of the reaction the profiles for butyl acetate derived from on-line direct liquid sampling mass spectrometry (DLSMS) differed from those of in-line MIR spectrometry owing to the 2 min transfer time between the reactor and mass spectrometer. As the reaction proceeded, however, the difference between the concentration profiles became less noticeable. DLSMS had advantages over in-line MIR spectrometry as it was easier to generate concentration profiles for all the components in the reaction. Also, it was possible to detect the presence of a simulated impurity of ethanol (at levels of 2.6 and 9.1% mol/mol) in butan-1-ol, and the resulting production of ethyl acetate, by DLSMS, but not by in-line MIR spectrometry.


Activated alpha2macroglobulin increases beta-amyloid (25-35)-induced toxicity in LAN5 human neuroblastoma cells.
The presence of the alpha2macroglobulin receptor/low density lipoprotein receptor-related protein (alpha2Mr/LRP) and its ligands alpha2macroglobulin (alpha2M), apoliprotein E, and plasminogen activators was detected in senile plaques of Alzheimer's disease (AD). To explore a possible role of alpha2M in neurodegenerative processes occurring in AD, we analyzed the effect of alpha2M on Abeta 25-35-induced neurotoxicity. Treatment of LAN5 human neuroblastoma cells with 10 microM beta-amyloid peptide fragment 25-35 (Abeta 25-35) for 72 h resulted in a 50% decrease in cell viability as determined by MTT incorporation and cell counts. The addition of alpha2M to the culture medium of these cells did not determine any effect, but when the activated form alpha2M* was used a dose-dependent decrease in cell viability was observed, the maximum effect being reached at 140 and 280 nM. Moreover, treatment of LAN5 cells with alpha2M* in combination with Abeta 25-35 increased the neurotoxicity of the amyloid peptide by 25%. This neurotoxic effect of alpha2M* seems to be related to its capability to bind and inactivate TGFbeta in the culture medium, since it was mimicked by a TGFbeta neutralizing antibody. A possible involvement of receptor-mediated endocytosis was ruled out, since alpha2M receptor is not present on LAN5, as revealed by RT-PCR and Western blotting experiments. The presence of alpha2M* in amyloid deposits of Alzheimer's disease has been recently reported and a possible impairment of LRP internalization processes has been hypothesized. Our data suggest that the local accumulation of alpha2M* in AD plaques may increase Abeta 25-35-induced neurotoxicity by neutralizing TGFbeta-mediated neuroprotective mechanisms.


Molecular biology of renal injury: emphasis on the role of the renin-angiotensin system.
The developments in molecular biology of the past decade have created a powerful technology with important, if not revolutionary, clinical applications. This review discusses the molecular biology of renal injury focusing on the renin-angiotensin system as a model, first considering the molecular physiology of the renin angiotensin system within the kidney and then considering its abnormalities in renal injury. All of the components of the renin-angiotensin system are present within the kidney and are involved in modulation of glomerular microcirculation, in proximal tubular reabsorptive function, in control of glomerular/tubular balance, in modulation of medullary blood flow, and in growth and repair of the renal tubule. A new understanding of these multiple roles of the renin-angiotensin system within the kidney is made feasible by combining physiological studies with techniques such as mRNA analysis (e.g., Northern and slot blots, in situ hybridization, and RNA protection assays), transgenic animal studies, transfection studies, and restriction fragment length polymorphism analysis. The ways in which such approaches have been used to examine the role of the renin-angiotensin system in acute renal failure, proteinuric states, renal hypertension, and diabetes mellitus are discussed.


Identification of key residues determining species differences in inhibitor binding of microsomal prostaglandin E synthase-1.
Microsomal prostaglandin E synthase-1 (MPGES1) is induced during an inflammatory reaction from low basal levels by pro-inflammatory cytokines and subsequently involved in the production of the important mediator of inflammation, prostaglandin E(2). Nonsteroidal anti-inflammatory drugs prevent prostaglandin E(2) production by inhibiting the upstream enzymes cyclooxygenases 1 and 2. In contrast to these conventional drugs, a new generation of NSAIDs targets the terminal enzyme MPGES1. Some of these compounds potently inhibit human MPGES1 but do not have an effect on the rat orthologue. We investigated this interspecies difference in a rat/human chimeric form of the enzyme as well as in several mutants and identified key residues Thr-131, Leu-135, and Ala-138 in human MPGES1, which play a crucial role as gate keepers for the active site of MPGES1. These residues are situated in transmembrane helix 4, lining the entrance to the cleft between two subunits in the protein trimer, and regulate access of the inhibitor in the rat enzyme. Exchange toward the human residues in rat MPGES1 was accompanied with a gain of inhibitor activity, whereas exchange in human MPGES1 toward the residues found in rat abrogated inhibitor activity. Our data give evidence for the location of the active site at the interface between subunits in the homotrimeric enzyme and suggest a model of how the natural substrate PGH(2), or competitive inhibitors of MPGES1, enter the active site via the phospholipid bilayer of the membrane.


Substrate promiscuity of an aminoglycoside antibiotic resistance enzyme via target mimicry.
The misuse of antibiotics has selected for bacteria that have evolved mechanisms for evading the effects of these drugs. For aminoglycosides, a group of clinically important bactericidal antibiotics that target the A-site of the 16S ribosomal RNA, the most common mode of resistance is enzyme-catalyzed chemical modification of the drug. While aminoglycosides are structurally diverse, a single enzyme can confer resistance to many of these antibiotics. For example, the aminoglycoside kinase APH(3')-IIIa, produced by pathogenic Gram-positive bacteria such as enterococci and staphylococci, is capable of detoxifying at least 10 distinct aminoglycosides. Here we describe the crystal structures of APH(3')-IIIa in complex with ADP and kanamycin A or neomycin B. These structures reveal that the basis for this enzyme's substrate promiscuity is the presence of two alternative subsites in the antibiotic binding pocket. Furthermore, comparison between the A-site of the bacterial ribosome and APH(3')-IIIa shows that mimicry is the second major factor in dictating the substrate spectrum of APH(3')-IIIa. These results suggest a potential strategy for drug design aimed at circumventing antibiotic resistance.


The Holo-Diagram. III: A Practical Device for Predicting Fringe Patterns in Hologram Interferometry.
The holo-diagram-a map of the setup used during exposure and reconstruction of a hologram-is a practical device for engineering uses of holography. It is designed to simplify the making and evaluating of holograms and to optimize the information retrieval in hologram interferometry. Existing complex interference patterns can, with the use of the holo-diagram, be interpreted with a minimum of calculations. The holo-diagram can also be used to predict fringe patterns caused by simple basic motions, e.g., translation and rotation. Prediction is by a direct method, with no calculations, and very quickly produces an approximate view of the patterns. Patterns predicted in this paper, and verified by experiment, in general correspond with those presented by other workers with some interesting differences due to the generality of the holo-diagram. It is also stated that the theory of the holo-diagram is relevant both for diffuse and specular reflection, and diagrams for pure translation of an object in the plane of the holo-diagram are given. A direct study of the parallax (dynamic method) is compared with a study of the apparent localization in depth (static method).


Polysaccharides from Radix Peucedani: Extraction, Structural Characterization and Antioxidant Activity.
In this study, an ultrasound-assisted green extraction method was applied for the extraction of polysaccharides from Radix Peucedani based on deep eutectic solvents (DESs), and the result showed that a DES system composed of betaine and 1,2-propylene glycol with a molar ratio of 1:2 possessed the optimal extraction efficiency for polysaccharides. Single-factor and Box-Behnken designs were used to determine the optimum extraction conditions for the maximum yields of polysaccharides from Radix Peucedani by using DESs. The maximum yields of polysaccharides attained 11.372% within a DES water content of 19%, an extraction time of 36 min, an extraction temperature of 54 °C, a solid-liquid ratio of 1:30 and an ultrasonic irradiation power of 420 W. The physicochemical properties of polysaccharides were analyzed using ICS and FT-IR, and the structure morphology was observed by SEM. The polysaccharides extracted from Radix Peucedani exhibited general antioxidant activities in vitro including DPPH, Hydroxyl and ABTS+ radical-scavenging activity. The antioxidant mechanism of Radix Peucedani polysaccharides was investigated using network pharmacology and molecular docking methods. The result showed that the high binding activity of glucose and IL1B, galactose and CASP3 was recognized as a potential mechanism for the antioxidant effects of Radix Peucedani polysaccharides.


Hesperidin and naringin attenuate hyperglycemia-mediated oxidative stress and proinflammatory cytokine production in high fat fed/streptozotocin-induced type 2 diabetic rats.
Abnormal regulation of glucose and impaired carbohydrate utilization that result from a defective or deficient insulin are the key pathogenic events in type 2 diabetes mellitus (T2DM). The present study was hypothesized to investigate the beneficial effects of hesperidin and naringin on hyperglycemia-induced oxidative damage in HFD/STZ-induced diabetic rats. Diabetes was induced by feeding rats with an HFD for 2 weeks followed by an intraperitoneal injection of STZ (35 mg/kg body weight). An oral dose of 50mg/kg hesperidin or naringin was daily given for 4 weeks after diabetes induction. At the end of the experimental period, blood was obtained from jugular vein and livers were rapidly excised and homogenized for biochemical assays. In the diabetic control group, levels of glucose, glycosylated hemoglobin (HbA1c%), MDA, NO, TNF-α and IL-6 were significantly increased, while serum insulin, GSH, vitamin C, and vitamin E levels were decreased. Both hesperidin and naringin administration significantly reversed these alterations. Moreover, supplementation with either compound significantly ameliorated serum and liver MDA, NO and glutathione, and liver antioxidant enzymes. Although detailed studies are required for the evaluation of the exact mechanism of the ameliorative effects of hesperidin and naringin against diabetic complications, these preliminary experimental findings demonstrate that both hesperidin and naringin exhibit antidiabetic effects in a rat model of T2DM by potentiating the antioxidant defense system and suppressing proinflammatory cytokine production.


Extracranial internal carotid artery dissections: noniatrogenic traumatic lesions.
Noniatrogenic traumatic extracranial internal carotid artery dissections were encountered in six patients (five men and one woman) 31 to 62 years old. All but one had overt cerebral ischemia manifest by paresis (three), sensory deficits (three), aphasia (three), or amaurosis fugax (two), One patient had an asymptomatic carotid artery bruit. Cerebral arteriography established the diagnosis in all cases. Internal carotid artery impingement between the mandible and transverse processes of the second and third cervical vertebrae, or undue stretching over these vertebral structures, were the most likely primary mechanisms of injury. Secondary complications, a result of distal thromboembolism, were evident in two patients. Direct cerebral revascularization, staged internal carotid artery constriction and ligation, as well as intensive nonoperative therapy were valid therapeutic options. There were no deaths. Treatment relieved transient ischemic symptoms or arrested progression of established neurologic deficits in each case. In select patients, early surgical intervention may lessen attending neurologic sequelae.


Malingering detected by forced choice testing of memory and tactile sensation: a case report.
A man with a possible diagnosis of multiple sclerosis was evaluated in connection with his claim for Social Security Disability. Two neurological examinations had revealed essentially normal findings. Neuropsychological testing revealed moderately severe deficits, but testing of his motivation with two forced choice measures, the Portland Digit Recognition Test and a finger graphesthesia procedure, both yielded performances significantly worse than chance. It was concluded that the patient was faking some or all of his deficits, and that his abilities had not been measured accurately. Forced choice testing is a robust procedure for documenting poor motivation.


Production of recombinant proteins in clonal root cultures using episomal expression vectors.
We have developed a fully contained system for expressing recombinant proteins that is based on clonal root cultures and episomal expression vectors. Clonal root lines expressing green fluorescent protein (GFP) or human growth hormone were generated from Nicotiana benthamiana leaves infected with the tobacco mosaic virus-based vector 30B after exposure to Agrobacterium rhizogenes. These lines accumulated GFP at over 50 mg per kg fresh tissue, a level that is comparable with other plant production systems in early stage development. Accumulation of both hGH and GFP in the clonal root lines was sustained over a 3-year period, and in the absence of antibiotic selection. This technology shows promise for commercial production of vaccine antigens and therapeutic proteins in contained facilities.


Risk factors for generally reduced productivity--a prospective cohort study of young adults with neck or upper-extremity musculoskeletal symptoms.
This study prospectively assessed the importance of individual conditions and computer use during school or work and leisure time as risk factors for self-reported generally reduced productivity due to musculoskeletal complaints among young adults with musculoskeletal symptoms in the neck or upper extremities. A cohort of 2914 young adults (18-25 years, vocational school and college or university students) responded to an internet-based questionnaire concerning musculoskeletal symptoms related to individual conditions and computer use during school or work and leisure time that possibly affected general productivity. Prevalence ratios (PR) were used to assess prospective risk factors for generally reduced productivity. The selected study sample (N=1051) had reported neck or upper-extremity symptoms. At baseline, 280 of them reported reduced productivity. A follow-up of the 771 who reported no reduced productivity was carried out after 1 year. Risk factors for self-reported generally reduced productivity for those followed-up were symptoms in two or three locations or dimensions for the upper back or neck and the shoulders, arms, wrists, or hands [PR 2.30, 95% confidence interval (95% CI) 1.40-3.78], symptoms persisting longer than 90 days in the shoulders, arms, wrists, or hands (PR 2.50, 95% CI 1.12-5.58), current symptoms in the shoulders, arms, wrists, or hands (PR 1.78, 95% CI 1.10-2.90) and computer use 8-14 hours/week during leisure time (PR 2.32, 95% CI 1.20-4.47). A stronger relationship was found if three or four risk factors were present. For women, a relationship was found between generally reduced productivity and widespread and current symptoms in the upper extremities. The main risk factors for generally reduced productivity due to musculoskeletal symptoms among young adults in this study were chronic symptoms in the upper extremities and widespread symptoms in the neck and upper extremities.


Temperature perturbation related to the invisible ink vibrationally excited nitric oxide monitoring (VENOM) technique: a simulation study.
The limits of applicability of the invisible ink variant of the vibrationally excited nitric oxide monitoring (VENOM) technique for three distinct flow fields is reported in this work. This technique involves the generation of a grid of vibrationally excited NO (X,Π2) by exciting the NO A-X electronic transition at 226 nm, which subsequently relaxes via fluorescence and collisional quenching to produce vibrationally excited NO (X,Π2). This grid is then probed by two laser sheets tuned to distinct rotational states. The resulting images allow for the simultaneous measurement of temperature and velocity. The flow fields presented in this work provide a range of NO concentrations, vibrational lifetimes, pressures, temperatures, and collisional quenching, which explore the applicability of the invisible ink variant to a wide range of conditions. We have modelled the initial NO, O2, and N2 vibrational and rotational energy distribution resulting from the combination of fluorescence and quenching of electronically excited NO. The subsequent rethermalization of the sample, in particular the long-time vibrational relaxation, has been modelled using a forced harmonic oscillator model. The time-dependent temperature perturbation due to the invisible ink technique is evaluated for two distinct timescales: a short-timescale temperature rise resulting from collisional quenching and rotational/translational thermalization and a long-timescale temperature rise caused by vibrational thermalization. Under low pressures where fluorescence dominates quenching, there is minimal temperature perturbation of the flow field on the timescale of a VENOM measurement, and the short-timescale temperature perturbation only becomes significant at high NO seed concentrations. The predicted signal-to-noise ratio of the invisible ink method is unaffected for low-pressure, low-temperature flow fields. However, preserving signal-to-noise ratio for a high-temperature, high-pressure flow field could prove challenging due to the impact of quenching and self-absorption. Overall, we find that the invisible ink method is predicted to be a viable laser-based diagnostic for velocimetry and thermometry over a wide range of experimental conditions.


Distinct type-1 protein phosphatases are associated with hepatic glycogen and microsomes.
The type-1 protein phosphatase associated with hepatic microsomes has been distinguished from the glycogen-bound enzyme in five ways. (1) The phosphorylase phosphatase/synthase phosphatase activity ratio of the microsomal enzyme (measured using muscle phosphorylase a and glycogen synthase (labelled in sites-3) as substrates) was 50-fold higher than that of the glycogen-bound enzyme. (2) The microsomal enzyme had a greater sensitivity to inhibitors-1 and 2. (3) Release of the catalytic subunit from the microsomal type-1 phosphatase by tryptic digestion was accompanied by a 2-fold increase in synthase phosphatase activity, whereas release of the catalytic subunit from the glycogen-bound enzyme decreased synthase phosphatase activity by 60%. (4) 95% of the synthase phosphatase activity was released from the microsomes with 0.3 M NaCl, whereas little activity could be released from the glycogen fraction with salt. (5) The type-1 phosphatase separated from glycogen by anion-exchange chromatography could be rebound to glycogen, whereas the microsomal enzyme (separated from the microsomes by the same procedure, or by extraction with NaCl) could not. These findings indicate that the synthase phosphatase activity of the microsomal enzyme is not explained by contamination with glycogen-bound enzyme. The microsomal and glycogen-associated enzymes may contain a common catalytic subunit complexed to microsomal and glycogen-binding subunits, respectively. Thiophosphorylase a was a potent inhibitor of the dephosphorylation of ribosomal protein S6, HMG-CoA reductase and glycogen synthase, by the glycogen-associated type-1 protein phosphatase. By contrast, thiophosphorylase a did not inhibit the dephosphorylation of S6 or HMG-CoA reductase by the microsomal enzyme, although the dephosphorylation of glycogen synthase was inhibited. The I50 for inhibition of synthase phosphatase activity by thiophosphorylase a catalysed by either the glycogen-associated or microsomal type-1 phosphatases, or for inhibition of S6 phosphatase activity catalysed by the glycogen-associated enzyme, was decreased 20-fold to 5-10 nM in the presence of glycogen. The results suggest that the physiologically relevant inhibitor of the glycogen-associated type-1 phosphatase is the phosphorylase a-glycogen complex, and that inhibition of the microsomal type-1 phosphatase by phosphorylase a is unlikely to play a role in the hormonal control of cholesterol or protein synthesis. Protein phosphatase-1 appears to be the principal S6 phosphatase in mammalian liver acting on the serine residues phosphorylated by cyclic AMP-dependent protein kinase.


Contraction Reserve With Ineffective Esophageal Motility on Esophageal High-Resolution Manometry is Associated With Lower Acid Exposure Times Compared With Absent Contraction Reserve.
Ineffective esophageal motility (IEM) is a minor motor disorder with potential reflux implications. Contraction reserve, manifested as augmentation of esophageal body contraction after multiple rapid swallows (MRS), may affect esophageal acid exposure time (AET) in IEM. Esophageal high-resolution manometry (HRM) and ambulatory reflux monitoring studies were reviewed over 2 years to identify patients with normal HRM, IEM (≥50% ineffective swallows), and absent contractility (100% failed swallows). Single swallows and MRS were analyzed using HRM software tools (distal contractile integral, DCI) to determine contraction reserve (mean MRS DCI to mean single swallow DCI ratio >1). Univariate analysis and multivariable regression analyses were performed to determine motor predictors of abnormal AET in the context of contraction reserve. Of 191 eligible patients, 57.1% had normal HRM, 37.2% had IEM, and 5.8% had absent contractility. Contraction reserve had no affect on AET in normal HRM. Nonsevere IEM (5-7 ineffective swallows) demonstrated significantly lower proportions with abnormal AET in the presence of contraction reserve (30.4%) compared with severe IEM (8-10 ineffective swallows) (75.0%, P = 0.03). Abnormal AET proportions in nonsevere IEM with contraction reserve (32.7%) resembled normal HRM (33.0%, P = 0.96), whereas that in severe IEM with (46.2%) or without contraction reserve (50.0%) resembled absent contractility (54.5%, P ≥ 0.6). Multivariable analysis demonstrated contraction reserve to be an independent predictor of lower upright AET in nonsevere (odds ratio 0.44, 95% confidence interval 0.23-0.88) but not severe IEM. Contraction reserve affects esophageal reflux burden in nonsevere IEM. Segregating IEM into severe and nonsevere cohorts has clinical value.


One Electron Changes Everything: Synthesis, Characterization, and Reactivity Studies of [Re(NCCH3)6]3.
Oxidation of [Re(NCCH3)6]2+ with the thianthrene radical cation results in the formation of [Re(NCCH3)6]3+, one of the very rare cases of a fully solvated +3 complex. It was fully characterized by spectroscopy and X-ray structure analysis. In contrast to its reduced analogue, [Re(NCCH3)6]3+ exhibits a much faster CH3CN exchange. Hence, substitution reactions proceed at 20 °C within minutes. Its potential as a versatile precursor for ReIII chemistry was examined with a series of substitution reactions. The more lipophilic analogue [Re(NCPh)6]3+ was synthesized by nitrile exchange in benzonitrile (NCPh). The Re(II) analogue of [Re(NCPh)6]3+, [Re(NCPh)6]2+, forms by AgI-mediated oxidation of in situ formed [Re(η6-C6H6)(NCPh3)3]+ in NCPh. The same synthetic strategy is feasible for the synthesis of [Re(NCCH3)6]2+ as well. [Re(NCCH3)6]3+ reacts with 1,4,7-trithiacyclononane (C6H12S3) to yield sevenfold-coordinated [Re(κ3-C6H12S3)2(NCCH3)]3+. The reaction of [Re(NCCH3)6]3+ with 1 equiv of (NBu4)X produces the ReIII monohalide complexes [ReX(NCCH3)5]2+ (X = Cl, Br, I). Mixed ReIII dihalides (trans-[ReXY(NCCH3)4]+) were obtained by treating [ReX(NCCH3)5]2+ with a second equivalent of (NBu4)Y (if X = Cl, Y = Br, I; if X = Br, Y = I). Because of this fast CH3CN exchange, [Re(NCCH3)6]3+ is a very suitable precursor for new ReIII complexes.


Contribution of salicylic acid glucosyltransferase, OsSGT1, to chemically induced disease resistance in rice plants.
Systemic acquired resistance (SAR), a natural disease response in plants, can be induced chemically. Salicylic acid (SA) acts as a key endogenous signaling molecule that mediates SAR in dicotyledonous plants. However, the role of SA in monocotyledonous plants has yet to be elucidated. In this study, the mode of action of the agrochemical protectant chemical probenazole was assessed by microarray-based determination of gene expression. Cloning and characterization of the most highly activated probenazole-responsive gene revealed that it encodes UDP-glucose:SA glucosyltransferase (OsSGT1), which catalyzes the conversion of free SA into SA O-beta-glucoside (SAG). We found that SAG accumulated in rice leaf tissue following treatment with probenazole or 2,6-dichloroisonicotinic acid. A putative OsSGT1 gene from the rice cultivar Akitakomachi was cloned and the gene product expressed in Escherichia coli was characterized, and the results suggested that probenazole-responsive OsSGT1 is involved in the production of SAG. Furthermore, RNAi-mediated silencing of the OsSGT1 gene significantly reduced the probenazole-dependent development of resistance against blast disease, further supporting the suggestion that OsSGT1 is a key mediator of development of chemically induced disease resistance. The OsSGT1 gene may contribute to the SA signaling mechanism by inducing up-regulation of SAG in rice plants.


On the origin of Hill's causal criteria.
The rules to assess causation formulated by the eighteenth century Scottish philosopher David Hume are compared to Sir Austin Bradford Hill's causal criteria. The strength of the analogy between Hume's rules and Hill's causal criteria suggests that, irrespective of whether Hume's work was known to Hill or Hill's predecessors, Hume's thinking expresses a point of view still widely shared by contemporary epidemiologists. The lack of systematic experimental proof to causal inferences in epidemiology may explain the analogy of Hume's and Hill's, as opposed to Popper's, logic.


Retrospective Validation of a 168-Gene Expression Signature for Glioma Classification on a Single Molecule Counting Platform.
Gene expression profiling has been shown to be comparable to other molecular methods for glioma classification. We sought to validate a gene-expression based glioma classification method. Formalin-fixed paraffin embedded tissue and flash frozen tissue collected at the Augusta University (AU) Pathology Department between 2000-2019 were identified and 2 mm cores were taken. The RNA was extracted from these cores after deparaffinization and bead homogenization. One hundred sixty-eight genes were evaluated in the RNA samples on the nCounter instrument. Forty-eight gliomas were classified using a supervised learning algorithm trained by using data from The Cancer Genome Atlas. An ensemble of 1000 linear support vector models classified 30 glioma samples into TP1 with classification confidence of 0.99. Glioma patients in TP1 group have a poorer survival (HR (95% CI) = 4.5 (1.3-15.4), p = 0.005) with median survival time of 12.1 months, compared to non-TP1 groups. Network analysis revealed that cell cycle genes play an important role in distinguishing TP1 from non-TP1 cases and that these genes may play an important role in glioma survival. This could be a good clinical pipeline for molecular classification of gliomas.


Viral status, immune microenvironment and immunological response to checkpoint inhibitors in hepatocellular carcinoma.
Immune checkpoint inhibitors (ICIs) targeting the programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathway have clinical activity in hepatocellular carcinoma (HCC), but only a subset of patients respond to these therapies, highlighting a need for novel biomarkers to improve clinical benefit. HCC usually occurs in the setting of liver cirrhosis from chronic hepatitis B or C viral infection, but the effects of viral status on the tumor immune microenvironment and clinical responses to ICIs in HCC remains unclear. We conducted a meta-analysis to estimate the objective response rates for PD-1/PD-L1 inhibitors in virally-infected and uninfected patients, and examined the effects of viral etiology on the tumor microenvironment using data from The Cancer Genome Atlas, as well as peripheral blood responses using an independent cohort of patients studied by mass cytometry (cytometry by time-of-flight (CyTOF)). Meta-analysis comparing objective response rates (ORR) between virally-infected and uninfected patients showed no clinically meaningful difference (absolute difference of ORR in virally-infected vs uninfected=-1.4%, 95% CI: -13.5% to 10.6%). There was no relationship between viral etiology on features of the tumor immune microenvironment that are known to modulate responses to PD-1/PD-L1 inhibitors, and the tumor mutational burden was similar between virally-infected and uninfected HCC. RNA sequencing of tissue-resident T cell and B cell repertoires similarly showed no effect of viral status on their diversity. CyTOF analysis of peripheral blood specimens further demonstrated similar expression of immune-related markers in response to PD-1 inhibitor therapy in virally-infected and uninfected HCC. There is no significant effect of viral etiology on the tumor immune microenvironment in HCC, and viral status should not be used as a criterion to select patients for PD-1/PD-L1 therapy.