Effects of Anthocleista djalonensis root extracts on reproductive hormones and testicular marker enzymes in adult male Wistar rats.
This study evaluated the effect of methanol and aqueous ethanol root extracts (200 mg/kg body weight) of Anthocleista djalonensis on sex hormone concentrations and testicular marker enzymes of adult rats after 60 days of administration followed by 60 days of treatment withdrawal. The results showed no significant changes in testosterone and follicle-stimulating hormone (FSH) levels during the 60 days of extract treatment. Interestingly, 60 days after treatment withdrawal, there was an increase in intratesticular and serum testosterone and serum FSH in the methanol but not aqueous ethanol extract post-treatment groups. Intratesticular 3β-hydroxysteroid dehydrogenase (3β-HSD) activity remained unaffected while that of 17β-HSD increased slightly during treatment of both extracts and the increase reached a statistical significance level (p < .05) during post-treatment. Gamma-glutamyltranspeptidase activity in the testis of the methanol but not aqueous ethanol extract-treated animals remained high during post-treatment compared to initial treatment values. Phytochemical analysis of the plant extracts showed that phenol and flavonoid constituents were higher in the methanol than the aqueous ethanol extract and has higher antioxidant activity. Altogether, post-treatment effect of the extract on the testis was more effective than treatment-related effect and the methanol extract appears to have better and consistent effects on the investigated parameters probably due to higher antioxidant activity conferred to it by its phenolic and flavonoid contents.


Progressive increase in peridevice leakage after the implantation of the watchman device on long-term serial echocardiographic follow-up.
Percutaneous left atrial appendage (LAA) occlusion using the Watchman device (Atritech, Plymouth, MN) is suggested as an alternative modality to warfarin for stroke prevention in patients with nonvalvular atrial fibrillation (AF). However, peridevice leakage resulting from incomplete LAA occlusion remains 1 of the most frequent limitations. We report a case of progressive increase in peridevice leakage after Watchman device implantation on long-term transesophageal echocardiographic follow-up accompanied by stroke.


The acoustic behavior of the Brazilian caatinga big rodent is incongruent to its actual position in Hydrochoerinae.
Our knowledge about the acoustic behavior of Caviinae species drove us to investigate Kerodon rupestris's (Caviidae: Hydrochoerinae) repertoire. This species is endemic to Caatinga and states as vulnerable in the Brazilian list of endangered species. We recorded sounds uttered by individuals from Santa Luzia, State of Paraíba, Brazil. We promoted interactions among 13 animals during intra and intersexual pairing sessions, under undisturbed interactions in captivity, and in free-living conditions. We found a repertoire of 13 call types, most of them (all except three) shared with Cavia and only five possibly shared also with Hydrochoerus: (1) Close contact and cohesion: tonal and noisy contact calls, tweet, slow-whistle; (2) Social regulation (incitement, subordination or auto-defense): whine, peepy-squeak, yelp; (3) Offensive aggression and status display: roar, snort, and teeth-chattering (4) Warning or intimidation: alarm-whistle, drrr, and drumming; (5) Courtship: purr and slow-whistle. The similarity of Kerodon signals to Hydrochoerinae species, despite the ecological differences, needs to be understood in comparative phylogenetic studies tracing back the origin of the courtship display in Caviidae. Thus, future research should focus on playback studies to test signals' biological function hypothesis.


Improved preparations of some per-O-acetylated aldohexopyranosyl cyanides.
3,4,6-Tri-O-acetyl-1,2-O-[1-(exo-, endo-cyano)ethylidene]-alpha-D- galacto- (1a/b), -alpha-D-gluco- (2a/b), and -beta-D-manno-pyranose (3a/b) were stereoselectively isomerized to the corresponding per-O-acetylated 1,2-trans-aldohexopyranosyl cyanides in 75, 16, and 62% yield, respectively, by treatment with boron trifluoride etherate in dry nitromethane. The corresponding per-O-acetylated 1,2-cis-aldohexopyranosyl cyanides were obtained concurrently in respective yields of 1.9, 0.9, and 4.8%. The per-O-acetylaldohexopyranosyl cyanide products were found stable to the reaction conditions and were readily isolated following completion of the rearrangement. It had previously been proved that reaction of 2,3,4,6-tetra-O-acetyl-alpha-D-manno- and -gluco-pyranosyl bromide with mercuric cyanide in nitromethane generates, in the ratio of approximately 1:1, the desired 1,2-trans-glycosyl cyanides and the corresponding 1,2-O-(1-cyanoethylidene) isomers (3a/b and 2a/b, respectively). Treatment of these reaction-mixtures with boron trifluoride etherate in nitromethane effected the rearrangement of 3a/b and 2a/b, thereby facilitating the isolation, and increasing the overall yields, of the per-O-acetylated 1,2-trans-D-manno- and -gluco-pyranosyl cyanides (58 and 30% total yield, respectively) relative to the earlier procedures. The boron trifluoride etherate-mediated reaction of per-O-acetyl-alpha- and -beta-D-galacto-, -alpha- and -beta-D-gluco-, -alpha-D-manno-, and -2-deoxy-2-phthalimido-beta-D-gluco-pyranoses with trimethylsilyl cyanide in nitromethane was also investigated. This reaction provides a "one-flask" synthesis of the corresponding per-O-acetylated 1,2-trans-aldohexopyranosyl cyanides in which 1,2-O-(1-cyanoethylidene) derivatives are isomerized in situ. Finally, improved preparations of the (not readily accessible) per-O-acetylated 1,2-cis-D-manno- and -gluco-pyranosyl cyanides are described. Thus, 2,3,4,6-tetra-O-acetyl-alpha- and -beta-D-mannopyranosyl cyanide (48 and 16% total yield, respectively) and -alpha- and -beta-D-glucopyranosyl cyanide (12 and 39% total yield, respectively) were synthesized by fusion of the corresponding alpha-D-glycosyl bromides with mercuric cyanide.


The comparison of point-of-care ultrasonography and radiography in the diagnosis of tibia and fibula fractures.
We aimed to compare the efficacy of Point-of-care ultrasonography (POCUS) with radiography in the diagnosis of tibia fracture (TF) and fibula fracture (FF), and determation of fracture characteristics. Patients aged 5-55 years who were admitted to ED due to low-energy, simple extremity trauma, who had a suspected TF and FF on physical examination were included in this prospective study. One physician performed POCUS examination. Other physician evaluated the radiography images. The obtained results were compared. A total of 62 patients were included in the study. TF was detected in 21 patients by radiography and in 24 patients by POCUS. FF was detected in 24 patients by radiography and in 25 patients by POCUS. Ten of the patients had both TF and FF. Compared with radiography, sensitivity, specificity, PPV and NPV of POCUS in the detection of TF were 100%, 93%, 88% and 100% (95% CI, 91-100%), respectively. Compared with direct X-ray imaging, sensitivity, specificity, PPV and NPV of POCUS in the detection of FF were 100%, 97%, 96% and 100% (95% CI, 96-100%), respectively. We determined that POCUS is also successful in detection of fracture features such as angulation, step-off, extension into the joint space that can determine the treatment decision. This study demonstrated that POCUS was found to be as successful as direct X-ray imaging in the diagnosis of TF and FF.


Variables associated with increased incidence of non-alcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes.
Type 2 diabetes (T2D) and non-alcoholic fatty liver disease (NAFLD) show a rapidly increasing incidence worldwide. Although both diseases often occur in the same patient population, their mutual influence is not fully understood. We therefore aimed at analyzing the impact of T2D on the incidence of NAFLD in a large cohort of outpatients in Germany. 32 201 patients with T2D diagnosed between 2012 and 2018 were identified in the IQVIA Disease Analyzer database. Probability of NAFLD was analyzed using Cox regression models. The cumulative incidence of NAFLD within the 7-year observation period was 4.3%. The probability of NAFLD was significantly higher among patients with T2D with increased body mass index but not hemoglobin A1c. Prescriptions of sodium-glucose cotransporter-2 inhibitors (HR: 0.54, 95% CI 0.45 to 0.64), glucagon-like peptide-1 receptor antagonists (HR: 0.65, 95% CI 0.52 to 0.81), and insulin (HR: 0.72, 95% CI 0.62 to 0.8) were significantly associated with lower incidence of NAFLD. Our data from a large population-based cohort of patients with T2D identified sociodemographic and therapeutic parameters associated with NAFLD incidence in patients with T2D which should be taken into account for novel therapeutic concepts.


[A simplified macrophage migration inhibition test to measure T-cell derived immunity in man].
A macrophage migration inhibition test using heterologous cells (DMMTH) is performed in an agarose-droplet technique. A suspension consisting of peritoneal exsudate cells from Guinea pig and human lymphocytes, mixed with 0,2 percent agarose is dispensed in droplets with a dispenser into culture chambers used for the leukocyte migration inhibition test. Macrophages migrate out of the gelified droplets. Migration areas can be measured by planimetry. Migration areas are exact to 8.6 percent. A significant migration inhibition, caused by migration inhibitory factors is observed after the reaction of the specific antigen with sensitized lymphocytes in the agarose droplet. The sensitivity of the test has been demonstrated in Mantoux positive persons.


Herpes simplex virus type 2 establishes latency in the mouse footpad and in the sensory ganglia.
Three mouse strains as well as wt and 7 ts mutants of herpes simplex virus (HSV) type 2 (strain HG52) have been used to investigate latency. The mice were inoculated in the right rear footpad. Virus reactivation following explanation and culture of the dorsal root ganglia and the footpad was scored. The results show that: HSV-2 can be maintained in the mouse footpad in a state indistinguishable from latency; virus gene functions necessary for latency can be identified by the use of ts mutants; and mouse strains differ in their ability to support latent infection. An infectious center assay was used to quantitate virus reactivation from dissociated dorsal root ganglia. HSV-1 strain 17 wt spread after inoculation at doses of greater than or equal to 5.0 X 10(5) plaque-forming units (pfu), producing latency also in contralateral ganglia but with lower efficiency.


Effects of D-baclofen and L-baclofen on the trigeminal nucleus.
D-Baclofen reduced the response to L-baclofen in the feline trigeminal nucleus, the spinal cord of the rat and in patients with trigeminal neuralgia, but not in slices of hippocampus or neocortex. The iontophoretic application of 10-20 nA L-baclofen depressed excitatory transmission in the trigeminal nucleus oralis, similar to the effect of 0.1-0.4 mg/kg L-baclofen, given intravenously. The concomitant iontophoresis of 10-20 nA D-baclofen reduced the effect of iontophoretically applied L-baclofen. However, larger doses of D-baclofen (30-60 nA) did not, while still larger doses (200-400 nA) by themselves depressed response of the neuron, similar to the action of small doses of L-baclofen. The iontophoresis of 30-40 nA L-baclofen had a stronger effect than that previously obtained with systemic administration and D-baclofen was not able to block it. These observations suggest that D-baclofen is a partial agonist at the GABAB receptor. Failure to observe a blocking effect of D-baclofen in slices of hippocampus or neocortex could be due to the larger doses used or to a difference in receptor types. The observations emphasise the need to test drugs at therapeutic concentrations in an appropriate model, in order to predict reliably their therapeutic actions.


[Detection of keratoconus based on a neural network with Orbscan].
To evaluate an artificial neural network in order to correctly identify Orbscan II tests of patients with normal and keratoconus corneas. A retrospective analysis included 98 Orbscan II tests of 59 subjects and an artificial neural network was created and trained based on the Java Neural Network 1.1 software. Seventy-three tests (59 normal tests and 14 keratoconus examinations) were applied to train the neural network and 25 eyes were used to test the method (19 normal eyes and 6 cases of keratoconus corneas). Backpropagation method was performed to train the neural network to 5% error and 0.2 learning rate. The trained neural network presented sensibility and specificity of 83 and 100% respectively. Artificial neural network can accurately help clinicians to classify keratoconus in Orbscan II tests.


Germ cell population of chick and quail embryos from DDT treated eggs.
Chick and Quail eggs were treated for 30 seconds, before incubation, with diluted aqueous suspension of commercial DDT. During the early stages of development, the germ cell population of treated embryos increases progressively and is only slightly lower than the control one. For each stage, the difference between the number of germ cells in controls and embryos contaminated by DDT, is not statistically significant. To exert its sterilizing effect on the germ cells, DDT must be administered before the beginning of incubation. At a later stage, when the gonocytes have colonized the gonads, the germ deficit becomes significant.


Epidemiology of Cholangiocarcinoma; United States Incidence and Mortality Trends.
Cholangiocarcinoma is an aggressive malignancy with few available studies assessing incidence and mortality. In this study, we aim to investigate trends of incidence and mortality in a large nation-wide epidemiologic study. We used SEER 18 database to study cholangiocarcinoma cases in the US during 2000-2015. Incidence and mortality rates of cholangiocarcinoma were calculated by race and were expressed by 1,000,000 person-years. Annual percent change (APC) was calculated using joinpoint regression software. We reviewed 16,189 patients with cholangiocarcinoma, of which 64.4% were intrahepatic. Most patients were whites (78.4%), males (51.3%), and older than 65 years (63%). A total of 13,121 patients died of cholangiocarcinoma during the study period. Cholangiocarcinoma incidence and mortality were 11.977 and 10.295 and were both higher among Asians, males, and individuals older than 65 years. Incidence rates have significantly increased over the study period (APC=5.063%, P<.001), while mortality increased significantly over the study period (APC=5.964%, P<.001), but decreased after 2013 (APC=-25.029, P<.001). The incidence and mortality of cholangiocarcinoma were increasing in the study period with significant observed disparities based on race and gender.


Bovine Endocarditis Associated with Mycoplasma bovis.
Mycoplasma bovis is a microorganism associated with pneumonia, mastitis, arthritis and otitis media of cattle; however, there are no reports of this organism causing bovine endocarditis. Five adult cattle with endocarditis characterized by caseated lesions (diameter 5-12 cm) of the endocardial surface of the left atrium, but without lesions in heart valves or affecting the right side of the heart, were identified in slaughterhouses in Japan. M. bovis was successfully isolated from the lesions and M. bovis antigen was detected immunohistochemically within the lesions. The results suggest that the lesions may have been associated with M. bovis alone. To our knowledge, this is the first demonstration of bovine endocarditis associated with M. bovis.


Prediction of prime editing insertion efficiencies using sequence features and DNA repair determinants.
Most short sequences can be precisely written into a selected genomic target using prime editing; however, it remains unclear what factors govern insertion. We design a library of 3,604 sequences of various lengths and measure the frequency of their insertion into four genomic sites in three human cell lines, using different prime editor systems in varying DNA repair contexts. We find that length, nucleotide composition and secondary structure of the insertion sequence all affect insertion rates. We also discover that the 3' flap nucleases TREX1 and TREX2 suppress the insertion of longer sequences. Combining the sequence and repair features into a machine learning model, we can predict relative frequency of insertions into a site with R = 0.70. Finally, we demonstrate how our accurate prediction and user-friendly software help choose codon variants of common fusion tags that insert at high efficiency, and provide a catalog of empirically determined insertion rates for over a hundred useful sequences.


EsD*5 gene frequency in Tuscany (Italy).
The distribution of the human red cell esterase D (EsD) "extended" polymorphism in a population sample from Tuscany (Italy) was studied using agarose gel isoelectric focusing. The estimated gene frequencies were: EsD*1 0.864, EsD*2 0.115, EsD*5 0.021. The EsD*5 allele frequency is very similar to those reported for other European populations. The "extension" of the EsD polymorphism may prove to be useful in paternity testing.


The relationship between loneliness and healthy aging indicators in Brazil (ELSI-Brazil) and England (ELSA): sex differences.
This study aimed to estimate five harmonized healthy aging indicators covering functional ability and intrinsic capacity among older women and men from Brazil and England and evaluate their association with loneliness. This was a cross-sectional study. We used two nationally representative samples of men and women aged ≥60 years from the Brazilian Longitudinal Study of Aging (ELSI-Brazil) wave 2 (2019-2021; n = 6929) and the English Longitudinal Study of Aging wave 9 (2018-2019; n = 5902). Healthy aging included five separate indicators (getting dressed, taking medication, managing money, cognitive function, and handgrip strength). Loneliness was measured by the 3-item University of California Loneliness Scale. Logistic regression models stratified by sex and country were performed. Overall, age-adjusted healthy aging indicators were worse in Brazil compared with England for both men and women. Considering functional ability, loneliness was negatively associated with all indicators (ranging from odds ratio [OR] = 0.26, [95% confidence interval (CI) 0.13-0.52] in English men regarding the ability to take medication to OR = 0.49 [95% CI 0.27-0.89] in Brazilian women regarding the ability to manage money). Considering intrinsic capacity, loneliness was negatively associated with a higher cognitive function (OR = 0.72; 95% CI 0.55-0.95 in English women) and a higher handgrip strength (OR = 0.61; 95% CI 0.45-0.83 in Brazilian women). Lonely women demonstrated lower odds of a higher number of healthy aging indicators than men in both countries. Country-specific social environments should be targeted by public policies to decrease loneliness and promote healthy aging later in life.


Dip-Coating Self-Assembly Fabrication and Polarization Sensitive Photoresponse of Aligned Single-Walled Carbon Nanotube Film.
It is challenging to obtain wafer-scaled aligned films for completely exploiting the promising properties of semiconducting single-walled carbon nanotubes (s-SWCNTs). Aligned s-SWCNTs with a large area can be obtained by combining water evaporation and slow withdrawal-induced self-assembly in a dip-coating process. Moreover, the tunability of deposition morphology parameters such as stripe width and spacing is examined. The polarized Raman results show that s-SWCNTs can be aligned in ±8.6°. The derived two terminal photodetector shows both a high negative responsivity of 41 A/W at 520 nm and high polarization sensitivity. Our results indicate that aligned films with a large area may be useful to electronics- and optoelectronics-related applications.


Individual differences in white matter microstructure of the face processing brain network are more differentiated from global fibers with increasing ability.
Face processing-a crucial social ability-is known to be carried out in multiple dedicated brain regions which form a distinguishable network. Previous studies on face processing mainly targeted the functionality of face-selective grey matter regions. Thus, it is still partly unknown how white matter structures within the face network underpins abilities in this domain. Furthermore, how relevant abilities modulate the relationship between face-selective and global fibers remains to be discovered. Here, we aimed to fill these gaps by exploring linear and non-linear associations between microstructural properties of brain fibers (namely fractional anisotropy, mean diffusivity, axial and radial diffusivity) and face processing ability. Using structural equation modeling, we found significant linear associations between specific properties of fibers in the face network and face processing ability in a young adult sample (N = 1025) of the Human Connectome Project. Furthermore, individual differences in the microstructural properties of the face processing brain system tended toward stronger differentiation from global brain fibers with increasing ability. This is especially the case in the low or high ability range. Overall, our study provides novel evidence for ability-dependent specialization of brain structure in the face network, which promotes a comprehensive understanding of face selectivity.


A novel method for the storage and transport of biological samples of therapeutic proteins prior to the detection of analytes using ELISA.
Therapeutic proteins have exhibited promising clinical applications in the diagnosis and treatment of some diseases. Prior to the detection of analytes using enzyme-linked immunosorbent assay, biological samples of therapeutic proteins are conventionally frozen at temperatures ranging from - 20 to - 80 °C to increase the stability of analytes. However, therapeutic proteins destabilization and aggregation may occur during the frozen storage or the freeze-thawing step. In this work, an effective method was proposed to freeze-dry therapeutic protein samples to allow subsequent storage or transport of samples without freezing them. This new method was validated with quality control samples of adalimumab and etanercept, and it was also used in the bioanalysis of adalimumab and etanercept in pharmacokinetic (PK) studies. Adalimumab and etanercept were stable for 14 days at 4 °C after being prepared and stored using the new method, with detection that was accurate and repeatable. Studies of adalimumab and etanercept in animals and humans showed that the PK parameters of the analytes stored with the new method were consistent with those of analytes stored using the conventional method. This effective method will be attractive for facilitating the storage and transport of plasma samples containing therapeutic proteins.


The "sickle-sign" in bone scintigraphy.
A side-effect of an intensive cytotoxic treatment on bone scanning will be reported: the so called "sickle sign", a diffuse activity around the calvarium seen only in patients at cytotoxic therapy. The differentiation between this harmless side-effect and abnormalities caused by the cancer could be done in our patients by a vertex view. All patients with skull metastases or meningeal carcinosis and this sign in the frontal and lateral view had an inhomogeneous uptake in the vertex view. This view should therefore be added to the standard views in all patients with the sickle sign.


Expression profile of toll-like receptor 2 mRNA in selected tissues of shark (Chiloscyllium sp.).
Sharks are a species of delight for immunologists from the evolutionary perspective since it is considered as the first species to have evolved the adaptive immune responses in addition to the innate immune system. One of the components of the highly conserved innate immune system is the toll-like receptors (TLR) which has a conserved overall protein structure throughout deuterostome evolution. There is no report that demonstrates the expression of these receptors in sharks. In this study we successfully amplified a 270 bp amplicon using a degenerate primer design strategy that corresponded to the Toll/IL-1 receptor (TIR) domain of TLR2 (GenBank ID: JF792813). BLAST analysis revealed a maximum nucleotide identity of 87% and 76% with the TLR2 of higher mammals and teleost fishes respectively. Domain prediction revealed a TIR structure between 1 and 87 amino acids that had a maximum identity of 58% and 76% with TLR2 - TIR protein of teleost fishes and higher mammals respectively. Phylogenetic analysis revealed a closer clustering of the shark TIR sequence with those from human, cattle, goat, sheep and chicken than with other fish species. Basal expression levels of the TLR2-TIR mRNA were found to be significantly higher in kidneys followed by fins, spleen and intestinal spiral valve (ISV). In tissues such as spleen and kidney the expression of the TLR2-TIR mRNA could be localized to lymphoid and macrophages like cells and tubular epithelial cells respectively. In-vivo exposure of sharks to peptidoglycan (TLR 2 ligand) resulted in 9 folds higher expression of TLR2-TIR mRNA in gills followed by 5 folds in the fins. However, when inoculated with a TLR ligand pool, the expression levels significantly increased to 12 fold in skin followed by epigonal, kidneys and ISV. These findings not only support the presence of the TLRs in sharks but also their induction upon exposure to specific ligands. Further studies are needed to identify their numbers, their ligand specificity and downstream cytokine responses.


Advances in Vaccine Adjuvants: Nanomaterials and Small Molecules.
Adjuvants have been extensively and essentially formulated in subunits and certain inactivated vaccines for enhancing and prolonging protective immunity against infections and diseases. According to the types of infectious diseases and the required immunity, adjuvants with various acting mechanisms have been designed and applied in human vaccines. In this chapter, we introduce the advances in vaccine adjuvants based on nanomaterials and small molecules. By reviewing the immune mechanisms induced by adjuvants with different characteristics, we aim to establish structure-activity relationships between the physicochemical properties of adjuvants and their immunostimulating capability for the development of adjuvants for more effective preventative and therapeutic vaccines.


Root Cause Analysis of Deaths in ST-Segment Elevation Myocardial Infarctions Treated With Primary PCI: What Can We Do Better?
Recent data demonstrate that mortality of patients with ST-elevation myocardial infarction (STEMI) has not changed despite dramatic reduction in door-to-balloon times. Identifying potential areas in care that can be further optimized to decrease mortality remains a priority. We performed a root cause analysis of all patients who died following primary percutaneous coronary intervention (PCI) during index hospitalization from 2008 to 2013 at the University of Michigan. Using a standardized data collection form, two interventional cardiologists and one non-invasive cardiologist reviewed patient care prior to arrival to the catheterization lab, while in the catheterization lab, and after primary PCI to determine cause of death and to rate potential preventability of death on a Likert scale (0 unpreventable - 4 mostly preventable). Of the 25 deaths over the 5-year period, 8 were deemed at least mildly preventable by one or more reviewer. No death was deemed totally preventable. Interreviewer agreement was moderate for both cause of death (nominal Krippendorff's alpha = .58) and preventability of death (nominal alpha = .233). In spite of this overall lack of agreement, in all 8 preventable cases at least one reviewer cited ischemia to balloon time as a potentially addressable factor associated with the death. Mortality following primary PCI was deemed mostly unpreventable. However, improvement in total ischemic time, and in particular symptom-onset to medical care, was identified as one potential target that might be of value in further reducing the mortality associated with STEMI.


Calcium-induced protein folding. Structure-affinity relationships in synthetic analogs of the helix-loop-helix calcium binding unit.
Three peptide analogs of the helix-loop-helix Ca2+ binding unit, 21-, 26-, and 34-residues in length, similar in sequence to rabbit skeletal troponin C site III have been prepared by the solid-phase method. The CD spectra of the 21-residue fragment indicated very little secondary structure in aqueous medium in the absence of Ca2+. Addition of Ca2+ increased the secondary structure of the peptide but the KCa was very weak, 3.1 x 10(2) M-1. The same peptide in hydrophobic medium in the absence of Ca2+ had considerable secondary structure and the KCa value increased considerably, 3.5 x 10(5) M-1. The 26-residue peptide, containing 5 more residues on the NH2 terminus of the 21-residue peptide, showed slightly more secondary structure in aqueous medium in the absence of Ca2+. Addition of Ca2+ to this peptide raised the amount of secondary structure in the metal ion-peptide complex and resulted in a higher KCa value, 3.8 x 10(4) M-1. By assuming that the COOH-terminal region of the 26-residue peptide-metal ion complex assumes a structure similar to that of the 21-residue peptide-metal ion complex, one is able to assign the increase in structure to the NH2-terminal side of the Ca2+-binding loop. Hydrophobic medium further increased the secondary structure of this peptide and also increased the KCa value to 4.5 x 10(5) M-1, a value similar to that obtained for the 21-residue peptide. The 34-residue peptide contained a further 8 amino acid residues on the NH2 terminus of the 26-residue peptide. This peptide had considerable secondary structure in aqueous medium which increased in the presence of Ca2+. The peptide has a reasonable affinity for Ca2+ in aqueous medium, KCa = 2.6 x 10(5) M-1. Again, a hydrophobic medium increased both the amount of secondary structure and the Ca2+ affinity constant, KCa = 9.2 x 10(5) M-1. A model of Ca2+-induced folding of the three peptides under different conditions is described and results obtained from this model are used to describe Ca2+ binding to the four Ca2+ binding units in rabbit skeletal troponin C.


Staphylococcus aureus ClpC ATPase is a late growth phase effector of metabolism and persistence.
Staphylococcus aureus Clp ATPases (molecular chaperones) alter normal physiological functions including an aconitase-mediated effect on post-stationary growth, acetate catabolism, and entry into death phase (Chatterjee et al., J. Bacteriol. 2005, 187, 4488-4496). In the present study, the global function of ClpC in physiology, metabolism, and late-stationary phase survival was examined using DNA microarrays and 2-D PAGE followed by MALDI-TOF MS. The results suggest that ClpC is involved in regulating the expression of genes and/or proteins of gluconeogenesis, the pentose-phosphate pathway, pyruvate metabolism, the electron transport chain, nucleotide metabolism, oxidative stress, metal ion homeostasis, stringent response, and programmed cell death. Thus, one major function of ClpC is balancing late growth phase carbon metabolism. Furthermore, these changes in carbon metabolism result in alterations of the intracellular concentration of free NADH, the amount of cell-associated iron, and fatty acid metabolism. This study provides strong evidence for ClpC as a critical factor in staphylococcal energy metabolism, stress regulation, and late-stationary phase survival; therefore, these data provide important insight into the adaptation of S. aureus toward a persister state in chronic infections.


Longitudinal study of plasma lipoproteins and hormones during pregnancy in normal and diabetic women.
Plasma lipoproteins were studied longitudinally at the 1st, 2nd, and 3rd trimester of gestation and at postpartum and postlactation in 12 age-matched PGDM women, 9 GDM women, and 12 healthy control subjects. FPG and HbA1c were higher in every case in PGDM women than in control subjects, whereas in GDM patients, glucose was augmented only after parturition. FFA and beta-hydroxybutyrate levels were higher in both PGDM and GDM patients than in control subjects during gestation but not after parturition. Total TGs and VLDL, LDL, and HDL TGs increased with gestational time in the three groups and declined at postpartum, and although total cholesterol and VLDL, LDL, and HDL cholesterol followed a similar trend, their rise was less pronounced, and the decline after parturition was slower than that of the TGs in the three groups, with no difference among them. The VLDL TG/cholesterol ratio declined in the three groups at the 3rd gestational trimester, whereas in both LDL and HDL, the TG/cholesterol ratio, but not the cholesterol/phospholipid ratio, increased during gestation in the three groups, indicating a specific enrichment of TGs in these particles. The increase in apoA-I and apoB with gestation was parallel to the respective changes in HDL and LDL cholesterol and, again, no difference was observed between the three groups. Plasma levels of beta-estradiol, progesterone, and prolactin increased sharply with gestation and declined at postpartum in the three groups, but absolute values of beta-estradiol and prolactin, at the three trimesters of gestation, were lower in PGDM patients, but progesterone levels were lower than controls in GDM women only at the 3rd trimester. (ABSTRACT TRUNCATED AT 250 WORDS)


Hematology and biochemistry of buffalo (Bubalus bubalis): influence of sex and age on reference values.
This research aimed to obtain a hematological and biochemical profile of buffalos (Bubalus bubalis) bred in a region of the Brazilian Legal Amazon. A total of 73 animals of mixed races, divided into three groups, were studied: group 1 (G1) comprised animals up to 11 months old; in group 2 (G2), subjects were 12 to 23 months old; and, in group 3 (G3), they were 24 months old and older. We performed the hematological analysis manually, and, for the biochemistry, we used biochemical analyzers. Age of the animals has influenced the monocyte count. The counting of these cells was higher (p > 0.05) in younger animals (G1). There was no difference (p > 0.05) in hematological parameters concerning sex. In regard to biochemistry, we found that age influenced results for albumin and urea. Animals from G1 had higher albumin values when compared to G2 and G3. Serum urea values were higher in animals from G3. Regarding the serum mineral parameters, there were no significant results when the different age groups and both sexes were compared. Thus, the hematological and biochemical values obtained can work as a reference for the bubaline species of animals bred in the studied region, under the same management and breeding conditions. Sex and age of the animals are necessary for the interpretation of the tests.


Sign Tracking in an Enriched Environment: A Potential Ecologically Relevant Animal Model of Adaptive Behavior Change.
When an object conditioned stimulus (CS) is paired with a food unconditioned stimulus (US), anticipatory goal-directed action directed at the US location (goal tracking) is accompanied by behavior directed at the object CS (sign tracking). Sign-tracking behavior appears to be compulsive and habit-like and predicts increased vulnerability to the addictive potential of drugs in animal models. A large body of the literature also suggests that environmental enrichment protects against the development of addiction-prone phenotypes. Thus, we investigated whether compulsive-like sign tracking develops in environmentally enriched rats trained directly in their enriched home environment. We demonstrate that adolescent enriched-housed male Sprague-Dawley rats readily sign track a 5% ethanol bottle CS in their home environment and at a rate higher than adolescent standard-housed rats. We also show that enriched adolescent rats sign track less than enriched adult-trained rats and that acute isolation stress affects sign- and goal-tracking performance of adolescents and adults differently. Sign tracking increased more in the adult than the adolescent rats. Whereas the younger rats showed a decrease in goal tacking after the final stressor manipulation, the adults showed increased goal tracking. Our results are consistent with recent studies, which suggest that although sign tracking performance is compulsive-like, it is not as inflexible and habit-like as previously assumed. Testing in an enriched home environment with object CSs having greater affordance than "neutral" lever CSs may provide greater ecological relevance for investigating the development and expression of adaptive and compulsive-like behaviors in translational research.


High-performance liquid chromatographic separation and determination of 2-(diethylamino)ethyl diphenylpropylacetate and its metabolites from biological samples.
A high-performance liquid chromatographic method for the determination of 2-(diethylamino)ethyl diphenylpropylacetate (SKF 525-A; Proadifen), its two deethylated metabolites and a hydrolytic metabolite in biological samples has been developed. From many solvent systems investigated the compounds are best separated on an ODS/Sil-X column using an acetonitrile-phosphate buffer system containing 0.2% decylamine. The use of acetonitrile overcomes limitations associated with previously published methods for separating basic compounds where decylamine was used in a methanolic eluent. With the sampling procedures used, overall recoveries from biological tissues were around 80%. The chromatographic system also separates three tricyclic antidepressants which can be used as internal standards.


Impact of the pre-examination phase on multicenter metabolomic studies.
The development of metabolomics in clinical applications has been limited by the lack of validation in large multicenter studies. Large population cohorts and their biobanks are a valuable resource for acquiring insights into molecular disease mechanisms. Nevertheless, most of their collections are not tailored for metabolomics and have been created without specific attention to the pre-analytical requirements for high-quality metabolome assessment. Thus, comparing samples obtained by different pre-analytical procedures remains a major challenge. Here, 1H NMR-based analyses are used to demonstrate how human serum and plasma samples collected with different operating procedures within several large European cohort studies from the Biobanking and Biomolecular Resources Infrastructure - Large Prospective Cohorts (BBMRI-LPC) consortium can be easily revealed by supervised multivariate statistical analyses at the initial stages of the process, to avoid biases in the downstream analysis. The inter-biobank differences are discussed in terms of deviations from the validated CEN/TS 16945:2016 / ISO 23118:2021 norms. It clearly emerges that biobanks must adhere to the evidence-based guidelines in order to support wider-scale application of metabolomics in biomedicine, and that NMR spectroscopy is informative in comparing the quality of different sample sources in multi cohort/center studies.


Use of the patient acceptable symptom state and the minimal clinically important difference to evaluate the outcomes of cataract extraction.
To determine the minimal clinically important difference and the patient acceptable symptom state for visual acuity and visual function, 2 key visual outcomes for patients undergoing cataract extraction, as an aid for evaluating the outcome of cataract extraction. Prospective cohort study. Multicenter study of 17 hospitals. A total of 4335 consecutive patients advised to undergo cataract extraction. Sociodemographic and clinical data, including visual acuity (VA), patient satisfaction, and responses to the Visual Function Index 14 (VF-14) and transitional questions were collected before and after cataract extraction. The patient acceptable symptom state and minimal clinically important difference were estimated for the entire sample and for subgroups by preintervention status and presence of ocular comorbidities. Among patients with simple cataract, postintervention patient acceptable symptom state values for VA ranged from 0.67 to 0.80, depending on preintervention VA, while VF-14 scores ranged from 88 to 90. For patients with any additional ocular comorbidity, VA ranged from 0.63 to 0.75, depending on preintervention VA, while VF-14 scores ranged from 86 to 92. For the entire sample, postintervention patient acceptable symptom state values were 0.75 for VA and 86.1 for VF-14 scores. Minimal clinically important difference for patients who reported being a little better ranged from 0.17 to 0.5 in VA and 0.41 to 37.46 in VF-14 scores, depending on preintervention status and presence of ocular comorbidities. The minimal clinically important difference and patient acceptable symptom state values are complementary parameters that may help in the clinical decision-making process by providing more meaningful estimates of the impact of cataract extraction on 2 important outcomes, visual acuity and visual function.


The relationship between return on investment and quality of study methodology in workplace health promotion programs.
To determine the relationship between return on investment (ROI) and quality of study methodology in workplace health promotion programs. Data were obtained through a systematic literature search of National Health Service Economic Evaluation Database (NHS EED), Database of Abstracts of Reviews of Effects (DARE), Health Technology Database (HTA), Cost Effectiveness Analysis (CEA) Registry, EconLit, PubMed, Embase, Wiley, and Scopus. Included were articles written in English or German reporting cost(s) and benefit(s) and single or multicomponent health promotion programs on working adults. Return-to-work and workplace injury prevention studies were excluded. Methodological quality was graded using British Medical Journal Economic Evaluation Working Party checklist. Economic outcomes were presented as ROI. ROI was calculated as ROI = (benefits - costs of program)/costs of program. Results were weighted by study size and combined using meta-analysis techniques. Sensitivity analysis was performed using two additional methodological quality checklists. The influences of quality score and important study characteristics on ROI were explored. Fifty-one studies (61 intervention arms) published between 1984 and 2012 included 261,901 participants and 122,242 controls from nine industry types across 12 countries. Methodological quality scores were highly correlated between checklists (r = .84-.93). Methodological quality improved over time. Overall weighted ROI [mean ± standard deviation (confidence interval)] was 1.38 ± 1.97 (1.38-1.39), which indicated a 138% return on investment. When accounting for methodological quality, an inverse relationship to ROI was found. High-quality studies (n = 18) had a smaller mean ROI, 0.26 ± 1.74 (.23-.30), compared to moderate (n = 16) 0.90 ± 1.25 (.90-.91) and low-quality (n = 27) 2.32 ± 2.14 (2.30-2.33) studies. Randomized control trials (RCTs) (n = 12) exhibited negative ROI, -0.22 ± 2.41(-.27 to -.16). Financial returns become increasingly positive across quasi-experimental, nonexperimental, and modeled studies: 1.12 ± 2.16 (1.11-1.14), 1.61 ± 0.91 (1.56-1.65), and 2.05 ± 0.88 (2.04-2.06), respectively. Overall, mean weighted ROI in workplace health promotion demonstrated a positive ROI. Higher methodological quality studies provided evidence of smaller financial returns. Methodological quality and study design are important determinants.


Urinary creatinine based equations for estimation of fat free mass in patients with intestinal insufficiency or intestinal failure.
Assessment of body composition is an important aspect of disease management in patients with intestinal insufficiency (INS) or intestinal failure (IF). However, in daily clinical settings most body composition methods are too expensive or impractical, leaving body composition to be assessed by less reliable methods such as skin fold thickness. The aim of this study was to investigate and validate the use of an equation for the estimation of fat-free mass (FFM) with bioelectrical impedance analysis (BIA) as reference method. A literature search for identification of urinary creatinine-based FFM-prediction equations was carried out a long side the creation of an equation by multiple linear regression. The correlation of each equation with FFM (measured by BIA in 277 patients with either INS or IF) was done by Pearson's correlation. Further investigation and validation of performance was done for the equations with the strongest correlation by Bland-Altman analysis, determination of root mean square error (RMSE), and intraclass correlation (ICC). The validation was carried out in a new group of 37 patients with either INS or IF. A total of 11 prediction equations were correlated with FFM measured by BIA. The equation called FFMmultiple and FFM-5 had the strongest correlation (r = 0.969, p < 0.01 and r = 0.950, p < 0.01, respectively). FFMmultiple was superior to FFM-5 regarding Bland-Altman analysis, RMSE, and ICC in the study group (Mean bias ± Standard Deviation = 0.042 ± 2.352 versus 0.309 ± 3.196; 95% limits of agreement = [-4.568; 4.651] versus [-5.955; 6.578]; RMSE = 0.158 versus 0.236; ICC = 0.969 versus 0.948). Cross-validation resulted in a Bland-Altman analysis with a statistically significant difference between FFMmultiple and FFM by BIA. FFM-5 showed wide 95% limits of agreement ([-6.977; 6.421]). Two urinary creatinine-based equations (FFMmultiple and FFM-5) showed promising results as possible substitutes to BIA, however further investigation and cross validation revealed inauspicious results. Thus, the present study cannot recommend the use of a prediction equation instead of BIA for the assessment of FFM in patients with INS and IF.


Chemical and olfactive impacts of organic matters on odor emission patterns from the simulated construction and demolition waste landfills.
The explosive increase of construction and demolition waste (CDW) caused the insufficient source separation and emergency disposal at domestic waste landfills in many developing countries. Some organic fractions were introduced to the CDW landfill process and resulted in serious odor pollution. To comprehensively explore the impacts of organic matters on odor emission patterns, five CDW landfills (OIL), with organic matters/ inert CDW components (O/I) from 5% to 30%, and the control group only with inert components (IL) or organics (OL) were simulated at the laboratory. The chemical and olfactive characters of odors were evaluated using the emission rate of 94 odorants content (ERtotal), theory odor concentration (TOCtotal), and e-nose concentration (ERENC), and their correlations with waste properties were also analyzed. It was found that the main contributors to ERtotal (IL: 93.0% NH3; OIL: 41.6% sulfides, 31.0% NH3, 25.9% oxygenated compounds) and TOCtotal (IL: 64.1% CH3SH, 28.2% NH3; OIL: 71.7% CH3SH, 24.8% H2S) changed significantly. With the rise of O/I, ERtotal, TOCtotal, and ERENC increased by 10.9, 20.6, and 2.1 times, respectively. And the organics content in CDW should be less than 10% (i.e., DOC<101.3 mg/L). The good regressions between waste properties (DOC, DN, pH) and ERENC- (r=0.86, 0.86, -0.88, p<0.05), TOCtotal- (r=0.82, 0.79, -0.82, p<0.05) implied that the carbon sources and acidic substances relating to organics degradation might result in that increase. Besides, the correlation analysis results (ERENC-vs.TOCtotal-, r=0.96, p<0.01; vs.ERtotal-, r=0.86, p<0.05) indicated that e-nose perhaps was a reliable odor continuous monitoring tool for CDW landfills.


Sophoraflavanone M, a prenylated flavonoid from Sophora flavescens Ait., suppresses pro-inflammatory mediators through both NF-κB and JNK/AP-1 signaling pathways in LPS-primed macrophages.
(2R)-3α,7,4'-trihydroxy-5-methoxy-8-(γ,γ-dimethylallyl)-flavanone is a prenylated flavonoid isolated from the anti-inflammatory herb Sophora flavescens Ait. We firstly named it sophoraflavanone M (SFM) in accordance with trivial names of related constitutes from this plant. Although various studies investigated the anti-inflammatory properties of prenylated flavonoids from Sophora flavescens Ait., that of SFM remains unclear and is yet to be determined. In the current study, we assessed the anti-inflammatory effects of SFM in LPS-induced in vivo and in vitro models. In the serum of endotoxemia mice, SFM significantly suppressed LPS-elevated inflammatory cytokines. Furthermore, at nontoxic concentrations, SFM reduced LPS-induced production of inflammatory mediators NO, IL-6, TNF-α, and MCP-1 in mouse primary peritoneal macrophages. Accordingly, in LPS-primed RAW264.7 cell line, it also inhibited these mediators' expression at both transcriptional and translational levels without cytotoxicity. Mechanistically, SFM is found to concurrently inhibit two important inflammatory signaling pathways, NF-κB and JNK/AP-1. SFM restrained phosphorylation and degradation of IκBα as well as the subsequent p65 translocation to dampen NF-κB activity. Meanwhile, it also suppressed JNK phosphorylation to inhibit the transcriptional activity of AP-1. These results provide material basis for traditional application of the anti-inflammatory herb Sophora flavescens Ait. and suggest SFM is a promising natural candidate for alleviating inflammatory conditions.


Carbon nanofiber-PEDOT composite films as novel microelectrode for neural interfaces and biosensing.
A clear need exists for novel nanostructured materials that are capable to meet the performance criteria of a number of neuronal therapies including neural recording, stimulation and sensing of bioactive molecules at the electrode-tissue interface. By combining Poly (3,4-ethylenedioxythiophene) (PEDOT), with Carbon Nanofibers (CNFs), we demonstrate a versatile approach for the synthesis of a novel composite material PEDOT:CNF with remarkable electrochemical properties, combining low impedance, high surface area, high charge injection capability and reliable neurotransmitters monitoring using amperometric techniques. The oxidized CNFs were utilized as dopants of PEDOT to prepare the composite coatings through electrochemical deposition on neural microelectrodes arrays (MEA). The PEDOT:CNF modified microelectrodes demonstrated the low specific impedance of 1.28 MΩ μm2 at 1 kHz and results in unrivalled charge injection limit of 10.03 mC/cm2 when compared to other reported organic electrode nanomaterials. Furthermore, amperometric detection performances were determined for the neurotransmitters dopamine and serotonin, exhibiting linear concentration range from 0.1 to 9 μM and from 0.06 to 9 μM respectively, high sensitivities (44.54 pA/nM.μm2 and 71.08 pA/nM.μm2, respectively) and low detection limits (0.045 μM and 0.056 μM, respectively). Cell viability was investigated on PEDOT:CNF coated microelectrodes to show that the composite material does not advocate any cytotoxicity. Taken together, these results suggest the great potential of PEDOT:CNF composite for developing next-generation multifunctional microelectrodes for applications in neural therapies.


Cytophotometry of post mortem glycogenolysis in different histochemical types of muscle fibres of the pig.
Serial transverse sections of porcine longissimus dorsi muscle (18 pigs, 50 to 178 kg live weight) were reacted for NAD tetrazolium reductase and ATPase at pH 9.4, and for glycogen with the periodic acid-Schiff (PAS) reaction. Three histochemical types of muscle fibre were identified; (1) strong ATPase and weak NADH oxidative activity; (2) strong ATPase and intermediate NADH oxidative activity; and (3) weak ATPase and strong NADH oxidative activity. Immediate post mortem samples from one side of each animal were compared with a later post mortem sample from the other side by measuring the absorbance of PAS-stained glycogen at 570 nm with a microscope photometer. Later post mortem absorbance was expressed as a percentage of immediate post mortem absorbance in each category of muscle fibre in order to compensate for distributional error and different starting levels of glycogen. Muscle fibres with weak ATPase and strong NADH oxidative activity showed a progressive decrease in absorbance of PAS-stained glycogen post mortem. In some animals, fibres with strong ATPase and intermediate or weak NADH oxidative activity showed an initial post mortem increase in absorbance of PAS-stained glycogen which was then followed by a progressive decrease. The maximum rates of decrease in absorbance in the three fibre types did not differ to any great extent.


Recent advances in the development of cyclin-dependent kinase 7 inhibitors.
Cyclin dependent kinase 7 (CDK7) plays a double role as it activates several other cyclin dependent kinases and participates to the initiation of transcription. This kinase is overexpressed in various types of tumors. Relatively few selective CDK7 inhibitors have been up to now disclosed. Most of these inhibitors belong to two chemical families: pyrazolopyrimidines and pyrazolotriazines on one side and pyrimidines on another side. They also differ by their molecular mechanism of action. Some are acting as competitive inhibitors and some others are covalent inhibitors. With these tools, the understanding of the potential therapeutic interest of CDK7 inhibitors in cancer is rapidly growing. They display antiproliferative activity against various types of tumors and leukemia and synergies have been identified. Two inhibitors are undergoing clinical testing. The most potent compounds inhibit a large number of cell-lines with IC50 < 200 nM.


Opportunities for improving pLDH-based malaria diagnostic tests.
Monoclonal antibodies to Plasmodium lactate dehydrogenase (pLDH) have been previously used to format immunochromatographic tests for the diagnosis of malaria. Using pLDH as an antigen has several advantages as a sensitive measure of the presence of parasites within patient blood samples. However, variable results in terms of specificity and sensitivity among different commercially available diagnostic kits have been reported and it has not been clear from these studies whether the performance of an individual test is due simply to how it is engineered or whether it is due to the biochemical nature of the pLDH-antibody reaction itself. A series of systematic studies to determine how various pLDH monoclonal antibodies work in combination was undertaken. Different combinations of anti-pLDH monoclonal antibodies were used in a rapid-test immunochromatographic assay format to determine parameters of sensitivity and specificity with regard to individual Plasmodium species. Dramatic differences were found in both species specificity and overall sensitivity depending on which antibody is used on the immunochromatographic strip and which is used on the colorimetric colloidal-gold used for visual detection. The results demonstrate the feasibility of different test formats for the detection and speciation of malarial infections. In addition, the data will enable the development of a universal rapid test algorithm that may potentially provide a cost-effective strategy to diagnose and manage patients in a wide range of clinical settings. These data emphasize that using different anti-pLDH antibody combinations offers a tractable way to optimize immunochromatographic pLDH tests.


Continued exposure of anti-cancer drugs to human iPS cell-derived cardiomyocytes can unmask their cardiotoxic effects.
Although the cardiotoxicity of anti-cancer drugs is an important issue, the underlying mechanisms remain unknown. To develop a sensitive assay system for cardiotoxicity, we examined effects of anti-cancer drugs on contractile functions of human iPS cell-derived cardiomyocytes by using non-invasive motion field imaging analysis with extended drug exposure time. We succeeded in continuously measuring stable contractile function. The continued exposure revealed that the difference in cardiotoxicity between cardiotoxic doxorubicin and less toxic erlotinib was more evident after 8 days of treatment than with 3 days of treatment, suggesting that continued exposure improved the predictive power for cardiotoxicity of anti-cancer drugs.


Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin nattokinase.
Here, we show that during in vivo folding of the precursor, the propeptide of subtilisin nattokinase functions as an intramolecular chaperone (IMC) that organises the in vivo folding of the subtilisin domain. Two residues belonging to β-strands formed by conserved regions of the IMC are crucial for the folding of the subtilisin domain through direct interactions. An identical protease can fold into different conformations in vivo due to the action of a mutated IMC, resulting in different kinetic parameters. Some interfacial changes involving conserved regions, even those induced by the subtilisin domain, blocked subtilisin folding and altered its conformation. Insight into the interaction between the subtilisin and IMC domains is provided by a three-dimensional structural model.


A Preliminary Report of COVID-19 in Children in India.
We describe the profile of COVID-19 in children from India in this multicentre observational study from tertiary care hospitals in West Bengal. Data of children up to 12 years presenting with positive results on SARS-CoV-2 RT-PCR test were included. The median (IQR) age of the 41 patients included was 1 (0.42-5.0) year. Eleven (26.8%) patients, including 6 neonates, never showed any symptoms. Fever was seen in only 9 patients (21%), and co-morbities were found in 61% of patients. There was one death.


Structures in Tetrahydrofolate Methylation in Desulfitobacterial Glycine Betaine Metabolism at Atomic Resolution.
Enzymes orchestrating methylation between tetrahydrofolate (THF) and cobalamin (Cbl) are abundant among all domains of life. During energy production in Desulfitobacterium hafniense, MtgA catalyzes the methyl transfer from methylcobalamin (Cbl-CH3 ) to THF in the catabolism of glycine betaine (GB). Despite its lack of sequence identity with known structures, we could show that MtgA forms a homodimeric complex of two TIM barrels. Atomic crystallographic insights into the interplay of MtgA with THF as well as analysis of a trapped reaction intermediate (THF-CH3 )+ reveal conformational rearrangements during the transfer reaction. Whereas residues for THF methylation are conserved, the binding mode for the THF glutamyl-p-aminobenzoate moiety (THF tail) is unique. Apart from snapshots of individual reaction steps of MtgA, structure-based mutagenesis combined with enzymatic activity assays allowed a mechanistic description of the methyl transfer between Cbl-CH3 and THF. Altogether, the THF-tail-binding motion observed in MtgA is unique compared to other THF methyltransferases and therefore contributes to the general understanding of THF-mediated methyl transfer.


A comprehensive overview of surgical and transcatheter therapies to treat tricuspid regurgitation in patients with heart failure.
The unique pathophysiologic considerations of severe tricuspid regurgitation (TR) have led to advancements in surgical and transcatheter treatments. The purpose of this review is to highlight the current surgical and transcatheter tricuspid valve interventions (TTVI) to functional TR. Surgical repair with ring annuloplasty consistently demonstrates better outcomes than surgical replacement or other repair approaches. However, surgical uptake of TR correction remains relatively low, and operative mortality rates are still high owing to multiple comorbidities and advanced tricuspid valve disease/right ventricular dysfunction at time of referral. Pivotal trials for tricuspid transcatheter edge-to-edge repair (T-TEER) and transcatheter TV replacement (TTVR) indicate improved quality of life compared to medical therapy alone for high-surgical-risk patients with severe symptomatic TR. Trials are underway to assess caval valve implantation (CAVI), which holds hope for many severe TR patients who are not ideal candidates for T-TEER or orthotopic TTVR. Peri-procedural optimization of right ventricular function remains critical to promote both device success and patient outcomes. Clinical outcomes after surgical TV intervention are poor, often due to intervening late in the disease course of TR. TTVI covers a treatment gap for patients deemed inoperable or high-surgical-risk, but earlier referral for TV interventions is still important prior to patients developing multiorgan dysfunction from chronic untreated TR.


The 'inductograph'--a graph describing the limits of the latent phase of induced labour in low risk situations.
The construction of an 'inductograph' is described which defines the rates of progress of the latent phase of labour in 80 per cent of a group of patients selected from a 'low risk' category. A standard induction regime was used, and changes in the uterine cervix were assessed quantitatively by a modified form of the Bishop scoring system.


Effects of stimulation intensity on intracranial cortico-cortical evoked potentials: A titration study.
The aim of the present study was to investigate the optimal stimulation parameters for eliciting cortico-cortical evoked potentials (CCEPs) for mapping functional and epileptogenic networks. We studied 13 patients with refractory epilepsy undergoing intracranial EEG monitoring. We systematically titrated the intensity of single-pulse electrical stimulation at multiple sites to assess the effect of increasing current on salient features of CCEPs such as N1 potential magnitude, signal to noise ratio, waveform similarity, and spatial distribution of responses. Responses at each incremental stimulation setting were compared to each other and to a final set of responses at the maximum intensity used in each patient (3.5-10 mA, median 6 mA). We found that with a biphasic 0.15 ms/phase pulse at least 2-4 mA is needed to differentiate between non-responsive and responsive sites, and that stimulation currents of 6-7 mA are needed to maximize amplitude and spatial distribution of N1 responses and stabilize waveform morphology. We determined a minimum stimulation threshold necessary for eliciting CCEPs, as well as a point at which the current-dependent relationship of several response metrics all saturate. This titration study provides practical, immediate guidance on optimal stimulation parameters to study specific features of CCEPs, which have been increasingly used to map both functional and epileptic brain networks in humans.


Observations relating to the efficiency of mixing in rapid reaction flow devices.
The principle of dimensional analysis of liquid flow has been applied to the problem of rapid mixing in flow apparatus. A model of the Hartridge-Roughton mixing chamber and observation tube has been scaled approximately 1/1.000 so that the times after mixing are approximately 6 s, the flow velocities are the order of 2 cm/s, and the distance from mixing to observation is 120 mm. Visual observation is employed to observe the end point of the mixing of acid-base with a color indicator, thymol blue. Quantitative estimates of the effect of the number of jets, the effect of screens placed in the flow stream, and the angle of jets, one with respect to another, lead to quantitative evaluations of these parameters for the scale model. The extent to which these parameters apply to the full-scale apparatus at faster flow velocities suggest that the general principles employed in the scale model are valid, although more emphasis is placed upon turbulence generation at the low flow velocities employed in this experiment than at the faster flow velocities employed in the full-scale apparatus.


Validation of Simultaneous Biochip-based Method for Screening of 3 Beta-Lactam Families Residues in Cow's Milk in Accordance with the European Union Decision 2002/657/EC and its Application on Real Samples.
Illegal and excessive use of veterinary antibiotics as a food additive for growth promotion in livestock can lead to allergic reactions and antibiotic resistance, which is a worldwide concern. A biochip-based semi-quantitative screening method of antimicrobial residues in milk was validated based on Commission Decision 2002/657/EC and the European guideline to validate screening methods for veterinary medicines. This multi-analytical screening method enables to determine of 3 beta-lactams (cefalexin, ampicillin, and cefuroxime) simultaneously. Analysis of 20 blank and 20 spiked milk samples showed that for all 3 antibiotic residues, the positivity threshold T was above cut-off value Fm, and no false-positive results were obtained for all 3 antibiotics. All detection capabilities (CCβ) were below Maximum Residue Level (MRL) authorized by European Commission. 47 UHT cow's milk samples collected from Tehran province, IR Iran, were screened, and compliance was found in 100% of samples. This study found that the biochip method is valid to determine antibiotic residues in milk samples at the measured validation levels. The method was fast, simple, and able to simultaneous screen three families of beta-lactams from a single milk sample with almost no sample preparation.


Antibiotic administration in targeted nanoparticles protects the faecal microbiota of mice.
Antibiotic therapy comes with disturbances on human microbiota, resulting in changes of bacterial communities and thus leading to well-established health problems. In this study, we demonstrated that targeted teicoplanin administration maintains the faecal microbiota composition undisturbed in a mouse model while reaching therapeutic improvements for S. aureus infection.


Dual effects of thyrotrophin-releasing hormone (TRH) on K+ conductance in frog pituitary melanotrophs. TRH-induced alpha-melanocyte-stimulating hormone release is not mediated through voltage-sensitive K+ channels.
Modulation of the activity of K+ channels by TRH and the possible involvement of this modulation in TRH-induced release of alpha-MSH were studied in cultured frog melanotrophs, using patch-clamp and perifusion techniques. Pars intermedia cells were enzymatically dispersed and cultured in Leibovitz medium. In order to test the viability of cultured cells, the amount of alpha-MSH released into the medium was measured by radioimmunoassay every day for 1 week of culture. The total amount of alpha-MSH released during the first 4 days of culture was 8.6 times higher than the intracellular content of alpha-MSH on day 1. Melanotrophs were identified by an indirect immunofluorescence technique using a specific antiserum to alpha-MSH. Recordings obtained in whole-cell, cell-attached and excised patch-clamp configurations showed that TRH induced a transient polarization concomitant with an increase in the probability of opening of Ca2+-activated K+ channels. This transient response was followed by a depolarization accompanied by an enhanced frequency of action potential discharge. TRH also induced a decrease in voltage-dependent K+ conductance. Application of tetraethylammonium, a K+ channel blocker, depolarized the cells and increased the basal secretory level without noticeable changes in TRH-evoked alpha-MSH release. These results demonstrate that the neuropeptide TRH both stimulates Ca2+-sensitive K+ channels and inhibits voltage-dependent K+ current in pituitary melanotrophs. Our data indicate that TRH-induced secretion of alpha-MSH is not a direct consequence of the lowering of K+ conductance. It thus appears that basal and TRH-induced alpha-MSH release occur through distinct pathways; the spontaneous release of alpha-MSH is probably linked to membrane potential, while modulation of the electrical activity is not directly involved in TRH-induced activation of the secretory process.


Technical note: Polyvinylpyrrolidone (PVP) and proteinase-K improve the efficiency of DNA extraction from Japanese larch wood and PCR success rate.
Illegal distribution of timber disrupts the timber market and depletes forest resources. DNA markers are used to verify the legal distribution of wood. However, it is difficult to obtain the quantity and quality of DNA suitable for genetic analysis because of the physicochemical properties of wood; therefore, an efficient wood DNA extraction method is required. In this study, to prepare an efficient DNA extraction method from Japanese larch (Larix kaempferi) wood, we investigated the ability of polyvinylpyrrolidone (PVP) and proteinase-K to improve DNA extraction efficiency and PCR success rate. It was found that the addition of PVP resulted in a significant increase in the DNA concentration of the treatment group compared to that of the control group, while the purity (A260/A280) showed no difference. Moreover, the addition of proteinase-K significantly increased both the DNA concentration and purity of the treatment group compared to those of the control group. Further analysis showed that the PCR success rate of psbC (approximately 350 bp) was higher than 90% in the control, PVP treatment, and proteinase-K treatment groups. However, in the PCR success rate of rbcL (approximately 1.3 kb) was higher in the proteinase-K and PVP treatment groups than in the control group. The addition of PVP and proteinase-K increased the success rate of PCR amplification for long regions by preventing DNA damage caused by phenolic compounds and proteins in the wood. The results of this study can thus develop DNA extraction methods to identify the species and origin of woods.


Periodontal dysbiosis associates with reduced CSF Aβ42 in cognitively normal elderly.
Periodontal disease is a chronic, inflammatory bacterial dysbiosis that is associated with both Alzheimer's disease (AD) and Down syndrome. A total of 48 elderly cognitively normal subjects were evaluated for differences in subgingival periodontal bacteria (assayed by 16S rRNA sequencing) between cerebrospinal fluid (CSF) biomarker groups of amyloid and neurofibrillary pathology. A dysbiotic index (DI) was defined at the genus level as the abundance ratio of known periodontal bacteria to healthy bacteria. Analysis of variance/analysis of covariance (ANOVA/ANCOVA), linear discriminant effect-size analyses (LEfSe) were used to determine the bacterial genera and species differences between the CSF biomarker groups. At genera and species levels, higher subgingival periodontal dysbiosis was associated with reduced CSF amyloid beta (Aβ)42 (P = 0.02 and 0.01) but not with P-tau. We show a selective relationship between periodontal disease bacterial dysbiosis and CSF biomarkers of amyloidosis, but not for tau. Further modeling is needed to establish the direct link between oral bacteria and Aβ.


Insomnia and stress of physicians during COVID-19 outbreak.
Healthcare workers are at high risk of developing sleep disorders during an outbreak. This study aimed to measure severity of sleep difficulty and its correlation with duration of deal with suspected/confirmed cases of novel coronavirus (COVID-19) in physicians. In this cross-sectional study, 268 physicians from different medical settings were included during the COVID-19 outbreak. The mean age and experience of physicians were 35.06 (33-70 years) and 10.13 years. The median duration of dealing with suspected/confirmed cases of COVID-19 was 1.0 (0-30 days). The mean sleep score and stress of physicians were 8.43 of 24.0 and 4.20 of 10, respectively. More than two-thirds of the physicians were sleepless (68.3%) and majority had stress (93.7%). The study did not find a significant difference in sleep score of physicians with different specialties (P = 0.059). However, most physicians were sleepless; including anesthesia and intensive care (77.8%); general physicians (80.8%), and obstetrics and gynecology (80.0%). They were sleepless in morning (58.7%); evening (77.8%); night (100%); and multi-shift (70.9%). The physicians who dealt with suspected or confirmed cases of COVID-19 or with stress had more escalated sleep compared to those who did not deal with patients or without stress (9.39 vs. 7.17 and 8.78 vs. 2.69 P < 0.001). The sleep of physicians was escalated with increasing stress (r = 0.558; P < 0.001) and a number of days that physicians dealt with suspected/confirmed cases of COVID-19 (r = 0.210; P = 0.001), respectively. The study confirmed that working with COVID-19 patients has a negative effect on the sleep of physicians.


Simultaneous recording with 30 microelectrodes in monkey visual cortex.
A 30-fold multielectrode for extracellular recording of neuronal spikes is described. Single neuronal spikes were isolated simultaneously by about half of the electrodes. The technique has been applied to demonstrate the spatial distribution of ocular dominance and orientation preference in striate cortex.


A case of drug-facilitated sexual assault by the use of flunitrazepam.
This article presents a case of drug-facilitated sexual assault on a female intoxicated with flunitrazepam. The male assailant added flunitrazepam (1 mg) to the female's soft drink, and had sexual intercourse with her while her consciousness was impaired. The complainant did not recall the events due to benzodiazepine-induced anterograde amnesia. The use of flunitrazepam was uncovered when its major metabolite, 7-amino flunitrazepam, was detected in a urine specimen collected when the complainant attended hospital approximately one day after consuming the adulterated drink.


A new mutation in the menin gene causes the multiple endocrine neoplasia type 1 syndrome with adrenocortical carcinoma.
Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant tumor syndrome that may be caused by mutations in the MEN1 gene on 11q13. Loss of function of the tumor suppressor gene MEN1 leads to synchronous or metachronous appearance of neuroendocrine tumors arising from neuroendocrine cells of the parathyroid and pituitary glands, the duodenum and pancreatic islets, and other endocrine organs such as the adrenal cortex. We here present a patient with MEN1 who developed hyperparathyroidism, multiple well differentiated functionally inactive neuroendocrine tumors of the pancreas and an adrenal carcinoma. We describe a new mutation at codon 443 in the coding region of exon 9 in the MEN1 gene, where a cytosine residue was exchanged for adenosine (TCC > TAC) and, consequently, serine for tyrosine (p.Ser443Tyr; c.1328C > A). [corrected] Also, we provide clinical data that may add to the genotype-phenotype discussion. We conclude that the novel mutation in the MEN1 gene described herein was clinically relevant.


A comparative overview of COVID-19, MERS and SARS: Review article.
Following the severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome coronavirus (MERS-CoV), a third, highly pathogenic coronavirus, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) appearing at end of 2019 led to a pandemic, increased panic and attracted global attention. This review analyzes the epidemiology, etiology, clinical characteristics, treatment and sequelae of the severe acute respiratory syndrome (SARS), the Middle East respiratory syndrome (MERS) and the 2019 novel coronavirus disease (COVID-19) to help provide direction for further studies that can help understand COVID-19.


Studying the impact of aging on memory systems: contribution of two behavioral models in the mouse.
In the present chapter, we describe our own attempts to improve our understanding of the pathophysiology of memory in aging. First, we tried to improve animal models of memory degradations occurring in aging, and develop common behavioral tools between mice and humans. Second, we began to use these behavioral tools to identify the molecular/intracellular changes occurring within the integrate network of memory systems in order to bridge the gap between the molecular and system level of analysis. The chapter is divided into three parts (i) modeling aging-related degradation in declarative memory (DM) in mice, (ii) assessing the main components of working memory (WM) with a common radial-maze task in mice and humans and (iii) studying the role of the retinoid cellular signaling path in aging-related changes in memory systems.


Advances in medical therapy for Crohn's disease.
Therapeutic research in Crohn's disease has been intensified in recent years. This has led to many novel approaches and insights into the mechanism of action of "classic" drugs. Antibiotics remain valuable but do not offer benefit when used in addition to corticosteroids. Immunomodulators remain the cornerstone for maintenance therapy, although certain corticosteroid-dependent patients can be switched to maintenance therapy with topical steroids. Azathioprine and 6-mercaptopurine remain efficient beyond 4 years in patients with relapses and elevated C-reactive protein in spite of this therapy. Infliximab has shown efficiency in maintenance of active and fistulizing Crohn's disease. In addition, "automatic reinfusion" was found to be superior to "on-demand" treatment. Infusion reactions and loss of response, most often caused by antibodies against infliximab, can be prevented with immunomodulators and corticosteroid infusions before dosing. Such alternative anti-tumor necrosis factor agents as adalimumab or CDP-870 may be less immunogenic. Other biologic agents, such as the anti-integrin monoclonal antibody natalizumab, were shown to be effective in maintaining remission and somewhat less so in induction of remission. Finally, much attention is being paid to alteration of the luminal flora with probiotics and helminth ova. Extracorporeal apheresis and even stem cell transplantation were found to be effective in isolated patients, but these therapies warrant further prospective and controlled investigation.


Exercise during pregnancy mitigates negative effects of parental obesity on metabolic function in adult mouse offspring.
Parental health influences embryonic development and susceptibility to disease in the offspring. We investigated whether maternal voluntary running during gestation could protect the offspring from the adverse effects of maternal or paternal high-fat diet (HF) in mice. We performed transcriptomic and whole-genome DNA methylation analyses in female offspring skeletal muscle and targeted DNA methylation analysis of the peroxisome proliferator-activated receptor-γ coactivator-1α (Pgc-1α) promoter in both male and female adult offspring. Maternal HF resulted in impaired metabolic homeostasis in male offspring at 9 mo of age, whereas both male and female offspring were negatively impacted by paternal HF. Maternal exercise during gestation completely mitigated these metabolic impairments. Female adult offspring from obese male or female parent had skeletal muscle transcriptional profiles enriched in genes regulating inflammation and immune responses, whereas maternal exercise resulted in a transcriptional profile similar to offspring from normal chow (NC)-fed parents. Maternal HF, but not paternal HF, resulted in hypermethylation of the Pgc-1α promoter at CpG-260, which was abolished by maternal exercise. These findings demonstrate the negative consequences of maternal and paternal HF for the offspring's metabolic outcomes later in life possibly through different epigenetic mechanisms, and maternal exercise during gestation mitigates the negative consequences.NEW & NOTEWORTHY Maternal or paternal obesity causes metabolic impairment in adult offspring in mice. Maternal exercise during gestation can completely mitigate metabolic impairment. Maternal obesity, but not paternal obesity, results in hypermethylation of the Pgc-1α promoter at CpG-260, which can be abolished by maternal exercise.


Genotype influences sulfur metabolism in broccoli (Brassica oleracea L.) under elevated CO2 and NaCl stress.
Climatic change predicts elevated salinity in soils as well as increased carbon dioxide dioxide [CO2] in the atmosphere. The present study aims to determine the effect of combined salinity and elevated [CO2] on sulfur (S) metabolism and S-derived phytochemicals in green and purple broccoli (cv. Naxos and cv. Viola, respectively). Elevated [CO2] involved the amelioration of salt stress, especially in cv. Viola, where a lower biomass reduction by salinity was accompanied by higher sodium (Na(+)) and chloride (Cl(-)) compartmentation in the vacuole. Moreover, salinity and elevated [CO2] affected the mineral and glucosinolate contents and the activity of biosynthetic enzymes of S-derived compounds and the degradative enzyme of glucosinolate metabolism, myrosinase, as well as the related amino acids and the antioxidant glutathione (GSH). In cv. Naxos, elevated [CO2] may trigger the antioxidant response to saline stress by means of increased GSH concentration. Also, in cv. Naxos, indolic glucosinolates were more influenced by the NaCl×CO2 interaction whereas in cv. Viola the aliphatic glucosinolates were significantly increased by these conditions. Salinity and elevated [CO2] enhanced the S cellular partitioning and metabolism affecting the myrosinase-glucosinolate system.


Caught red handed: modeling and confirmation of the myeloperoxidase ceruloplasmin alpha-thrombin complex.
The work is devoted to the study of the structural characteristics of the myeloperoxidase-ceruloplasmin-thrombin complex using small-angle neutron scattering methods in combination with computer modeling, as well as surface plasmon resonance and solid-phase enzyme assay. We have previously shown that the functioning of active myeloperoxidase during inflammation, despite the presence in the blood of an excess of ceruloplasmin which inhibits its activity, is possible due to the partial proteolysis of ceruloplasmin by thrombin. In this study, the myeloperoxidase-ceruloplasmin-thrombin heterohexamer was obtained in vitro. The building of a heterohexamer full-atomic model in silico, considering the glycosylation of the constituent proteins, confirmed the absence of steric barriers for the formation of protein-protein contacts. It was shown that the partial proteolysis of ceruloplasmin does not affect its ability to bind to myeloperoxidase, and a structural model of the heterohexamer was obtained using the small-angle neutron scattering method.


DDA3 associates with MCAK and controls chromosome congression.
DDA3 regulates spindle microtubule (MT) dynamics and chromosome movement in mitosis through its interaction with and subsequent recruitment of Kif2a, a minus end-MT depolymerase. Depletion of DDA3 causes a hyper-stabilization of spindle MT, a loss of inter-kinetochore tension, and a defect in chromosome congression, leading to unaligned chromosomes at metaphase. We report here that DDA3 is also localized at kinetochores and interacts with MCAK. Furthermore, CENP-E, a plus end-motor protein, accumulates at kinetochores in unaligned chromosomes in mitotic cells depleted of DDA3. On the other hand, the localization of chromosomal passenger complex (CPC) and the kinase activity of Aurora B are normal in DDA3-depleted cells. We conclude that MCAK and CENP-E are involved in DDA3-mediated chromosome congression.


Nigrostriatal rAAV-mediated GDNF overexpression induces robust weight loss in a rat model of age-related obesity.
Intraventricular administration of glial cell line-derived neurotrophic factor (GDNF) in primate and humans to study Parkinson's disease (PD) has revealed the potential for GDNF to induce weight loss. Our previous data indicate that bilateral continuous hypothalamic GDNF overexpression via recombinant adeno-associated virus (rAAV) results in significant failure to gain weight in young rats and weight loss in aged rats. Based on these previous results, we hypothesized that because the nigrostriatal tract passes through the lateral hypothalamus, motor hyperactivity mediated by nigrostriatal dopamine (DA) may have been responsible for the previously observed effect on body weight. In this study, we compared bilateral injections of rAAV2/5-GDNF in hypothalamus versus substantia nigra (SN) in aged Brown-Norway X Fisher 344 rats. Nigrostriatal GDNF overexpression resulted in significantly greater weight loss than rats treated in hypothalamus. The nigral or hypothalamic GDNF-induced weight loss was unrelated to motor activity levels of the rats, though some of the weight loss could be attributed to a transient reduction in food intake. Forebrain DA levels did not account for the observed effects on body weight, although GDNF-induced increases in nucleus accumbens DA may have partially contributed to this effect in the hypothalamic GDNF-treated group. However, only nigrostriatal GDNF overexpression induced activation of phosphorylated extracellular signal-regulated kinase (p-ERK) in a small population of corticotrophin-releasing factor [corticotrophin-releasing hormone (CRH)] neurons located specifically in the medial parvocellullar division (MPD) of the paraventricular nucleus of the hypothalamus. Activation of these hypothalamic CRH neurons likely accounted for the observed metabolic effects leading to weight loss in obese rats.


Metabolic syndrome and its components are related to psychological disorders: A population based study.
Psychological disorders are considered as today's one of the most important public health problems all around the world. Another alarming condition, which attributed to cardiovascular disease (CVD) is metabolic syndrome (Mets). The aims of the current study were to explore (1) the prevalence of anxiety and depression in a large representative population of Iran, and (2) evaluate their possible association with MetS patients. Applying a randomized stratified-cluster approach, 9829 adults, aged 35-65 years, were recruited as part of the MASHAD study. Socio demographic, anthropometric and biochemical analyses were performed. Beck's anxiety inventory and Beck's depression inventory II were applied to assessment of the degree of anxiety and depression among the subjects. The prevalence of MetS, anxiety and depression were 38.8%, 25% and 31.5%, respectively. The prevalence of these factors was significantly (P<0.05) higher in women, compared to men and subjects with MetS were significantly (P<0.05) associated with both anxiety and depression. Additionally, multivariate analysis after adjusting for potential confounders showed a significant relationship between MetS and anxiety (p=0.04). Our data demonstrates the significant association of MetS components with anxiety and depression and high prevalence of these co-morbidities in adults, particularly in women.


Crowdsourced assessment of surgical skills: A systematic review.
Crowdsourced assessment utilizes a large group of untrained individuals from the general population to solve tasks in the medical field. The aim was to examine the correlation between crowd workers and expert surgeons for the use of crowdsourced assessments of surgical skills. A systematic literature review was performed on April 14th, 2021 from inception to the present. Two reviewers screened all articles with eligibility criteria of inclusion and assessed for quality using The Medical Education Research Study Quality Instrument (MERSQI) and Newcastle-Ottawa Scale-Education (NOS-E)(Holst et al., 2015).7General information was extracted for each article. 250 potential studies were identified, and 32 articles were included. There appeared to be a generally moderate to very strong correlation between crowd workers and experts (Cronbach's alpha 0.72-0.95, Pearson's r 0.7-0.95, Spearman Rho 0.7-0.89, linear regression 0.45-0.89). Six studies had either questionable or no significant correlation between crowd workers and experts. Crowdsourced assessment can provide accurate, rapid, cost-effective, and objective feedback across different specialties and types of surgeries in dry lab, simulation, and live surgeries.


Genome-wide family-based study in torus palatinus affected individuals.
Tori or exostoses are bony growths that appear in different oral regions. Torus palatinus, more specifically, develop in the palate midline and can impair proper word pronunciation and hinder the fabrication and use of dentures. Even though a multifactorial inheritance model has been suggested for torus palatinus appearance, precise genetic factors involved in its etiology remain unclear. Hence, in this study we aimed to identify variants across the genome of individuals from 46 Filipino families that associate with torus palatinus. All families were composed of fishermen or landless rural dwellers who provided blood samples for DNA extraction and genotyping. A total of 3519 single nucleotide polymorphisms (SNPs) were analyzed through a transmission disequilibrium test in individuals affected by torus palatinus and their unaffected family members. Fourteen SNPs showed trends for associations to the level of p < .005 threshold and several others were nominally (p < .05) associated with torus palatinus. We highlight SNP rs6582285, which is located in the CAPS2 gene, being the C allele less transmitted than the T allele in our sample. The C allele of CAPS2 rs6582285 protects from having torus palatinus whereas the other associations found were linked to an increased risk of developing the condition. Trends for associations were identified for several markers across the genome, supporting the hypothesis that torus palatinus has a multifactorial mode of inheritance. We hope that our study contributes to a better understanding of torus palatinus etiology and helps guide future research in examining genes for this often-overlooked condition in different populations.


Physical Health, Media Use, and Mental Health in Children and Adolescents With ADHD During the COVID-19 Pandemic in Australia.
To examine the impact of COVID-19 restrictions among children with attention-deficit/hyperactivity disorder (ADHD). Parents of 213 Australian children (5-17 years) with ADHD completed a survey in May 2020 when COVID-19 restrictions were in place (i.e., requiring citizens to stay at home except for essential reasons). Compared to pre-pandemic, children had less exercise (Odds Ratio (OR) = 0.4; 95% CI 0.3-0.6), less outdoor time (OR = 0.4; 95% 0.3-0.6), and less enjoyment in activities (OR = 6.5; 95% CI 4.0-10.4), while television (OR = 4.0; 95% CI 2.5-6.5), social media (OR = 2.4; 95% CI 1.3-4.5), gaming (OR = 2.0; 95% CI 1.3-3.0), sad/depressed mood (OR = 1.8; 95% CI 1.2-2.8), and loneliness (OR = 3.6; 95% CI 2.3-5.5) were increased. Child stress about COVID-19 restrictions was associated with poorer functioning across most domains. Most parents (64%) reported positive changes for their child including more family time. COVID-19 restrictions were associated with both negative and positive impacts among children with ADHD.


Iodine-125 implants for carcinoma of the prostate.
One hundred-thirteen patients underwent Iodine-125 prostate implant and lymphadenectomy at Yale-New Haven Hospital from 1974 through 1980. The distribution by clinical stage was: 7 Stage A2, 86 Stage B, and 20 Stage C patients. Ninety-four patients had a negative lymphadenectomy (N-) and 19 patients (17%) had metastatic disease in the pelvic lymph nodes (N+). The actuarial 5-year survival for all 113 patients was 87% (+/- 6%: 95% confidence limits). Sixty-five percent of our 113 patients are disease free (NED) from 2 to 9 years following implant. Sixty-seven (N-) patients with clinical Stage B disease, whose tumors were either well differentiated or moderately well differentiated, have an actuarial 5-year NED survival of 84% (+/- 8%). Twenty (N-) patients with either clinical Stage C disease or poorly differentiated tumors have an actuarial 5-year NED survival of only 31% (+/- 20%). For the 19 (N+) patients, the actuarial 5-year NED survival is 38% (+/- 22%). Local tumor control was 85% for all Stage B patients and 75% for all Stage C patients. Only 10 patients (9%) have developed long-term gastrointestinal or genitourinary complications following their implant. Iodine-125 implant appears to be a reasonable alternate form of therapy in highly selected groups of patients with carcinoma of the prostate.


Laser threshold and cell damage mechanism for intravascular photoacoustic imaging.
Intravascular photoacoustic (IVPA) imaging is being developed to image atherosclerotic plaques, a leading cause of morbidity and mortality in the United States. However, the safety of this imaging modality, which requires repeated irradiation with short laser pulses, has not yet been investigated. This study has two objectives. First, determine in vitro the limit of cumulative fluence that can be applied to cells before death at IVPA relevant wavelengths. Second, evaluate if high single pulse fluences are a potential cause of cell death during IVPA imaging. Experiments were conducted using endothelial cells, macrophages, and smooth muscle cells. The cumulative fluence experiments were conducted at 1064 and 1197 nm, using a high pulse repetition frequency laser. Cells were irradiated with a wide range of cumulative fluences and evaluated for cell death. The thresholds for death were compared to the maximum expected clinical cumulative fluence. To evaluate the effect of single pulse fluences, cells were irradiated at 1064, 1210, and 1720 nm. Light was delivered at a range of pulse energies to emulate the fluences that cells would be exposed to during clinical IVPA imaging. At 1064 nm, all three cell types remained viable at cumulative fluences above the maximum expected clinical cumulative fluence, which is calculated based on common IVPA imaging protocols. At 1197 nm, cells were viable near or just below the maximum expected clinical cumulative fluence, with some cell type to cell type variation. All three cell types remained viable after irradiation with high single pulse fluences at all three wavelengths. The cumulative fluence experiments indicate that safety considerations are likely to put constraints on the amount of irradiation that can be used in IVPA imaging protocols. However, this study also indicates that it will be possible to use IVPA imaging safely, since cumulative fluences could be reduced by as much as two orders of magnitude below the maximum expected clinical cumulative fluence by varying the imaging protocol, albeit at the expense of image quality. The single pulse fluence experiments indicate that cell death from single pulse fluence is not likely during IVPA imaging. Thus, future studies should focus on heat accumulation as the likely mechanism of tissue damage. Lasers Surg. Med. 51:466-474, 2019. © 2018 Wiley Periodicals, Inc.


Quantitative sensory testing measures individual pain responses in emergency department patients.
Refining and individualizing treatment of acute pain in the emergency department (ED) is a high priority, given that painful complaints are the most common reasons for ED visits. Few tools exist to objectively measure pain perception in the ED setting. We speculated that variation in perception of fixed painful stimuli would explain individual variation in reported pain and response to treatment among ED patients. In three studies, we 1) describe performance characteristics of brief quantitative sensory testing (QST) in 50 healthy volunteers, 2) test effects of 10 mg oxycodone versus placebo on QST measures in 18 healthy volunteers, and 3) measure interindividual differences in nociception and treatment responses in 198 ED patients with a painful complaint during ED treatment. QST measures adapted for use in the ED included pressure sensation threshold, pressure pain threshold (PPT), pressure pain response (PPR), and cold pain tolerance (CPT) tests. First, all QST measures had high inter-rater reliability and test-retest reproducibility. Second, 10 mg oxycodone reduced PPR, increased PPT, and prolonged CPT. Third, baseline PPT and PPR revealed hyperalgesia in 31 (16%) ED subjects relative to healthy volunteers. In 173 (88%) ED subjects who completed repeat testing 30 minutes after pain treatment, PPT increased and PPR decreased (Cohen's dz 0.10-0.19). Verbal pain scores (0-10) for the ED complaint decreased by 2.2 (95% confidence intervals [CI]: 1.9, 2.6) (Cohen's dz 0.97) but did not covary with the changes in PPT and PPR (r=0.05-0.13). Treatment effects were greatest in ED subjects with a history of treatment for anxiety or depression (Cohen's dz 0.26-0.43) or with baseline hyperalgesia (Cohen's dz 0.40-0.88). QST reveals individual differences in perception of fixed painful stimuli in ED patients, including hyperalgesia. Subgroups of ED patients with hyperalgesia and psychiatric history report larger treatment effects on ED pain and QST measures.


Cervical carcinoid ("argyrophil cell" carcinoma) associated with an endocervical adenocarcinoma: a light and ultrastructural study.
On microscopic examination of a cervical carcinoid removed from a 29-year-old woman, foci of endocervical adenocarcinoma in situ (AIS) and severe squamous dysplasia were present immediately adjacent to the invasive carcinoid tumor. Cells of the carcinoid tumor contained abundant intracytoplasmic argyrophilic granules and numerous neurosecretory granules (NSG) on ultrastructural study. Neoplastic cells of the endocervical AIS were devoid of argyrophilic granules and contained no NSGs. The present neoplastic process appeared similar to cases of a "mixed" type neoplasm composed of a carcinoid tumor and an adenocarcinoma, which have been reported from several areas of the body but only once previously from the cervix. The possible mechanisms accounting for such an unusual neoplastic process are discussed.