Physicians' Attitudes on Management of Osteopenia in South Korea.
Osteopenia patients have a risk of fracture and may develop osteoporosis. We investigated physicians' management of osteopenia patients in South Korea. A survey was conducted using a questionnaire including 6 items: (1) do you think anti-osteoporosis medications are necessary in osteopenia patients?; (2) what is your preference to manage osteopenia patients, except for anti-osteoporosis medications?; (3) what is your indication for the anti-osteoporosis medication in osteopenia patients?; (4) what kind of anti-osteoporosis medication do you prefer to treat osteopenia patients?; (5) do you use bisphosphonates?; and (6) if not, what is the reason for not using?. Among the 173 participants, 150 (86.7%) replied that anti-osteoporosis medications were necessary in osteopenia patients. Indications for the medication were (1) past medical history of pathologic fracture in 85 (49.1%); (2) T-score <-2.5 on dual energy X-ray absorptiometry in 73 (42.2%); (3) previous history of osteoporosis in 44 (25.4%); (4) risk of fracture according to fracture risk assessment tool in 34 (19.7%); and (5) progressive bone loss in 31 (17.9%). One hundred and sixteen (67.1%) favored bisphosphonates, 93 (53.8%) selective estrogen-receptor modulator, and 24 (13.9%) hormone replacement therapy. Thirty-one (17.9%) replied that they do not use bisphosphonates due to (1) restricted reimbursement by the health insurance in 24 (77.4%); and (2) bisphosphonate-related complications in 19 (61.3%). Most respondents (86.7%) thought anti-osteoporosis medications were necessary in osteopenia patients, but 17.9% of the respondents did not use bisphosphonates. Restricted reimbursement by the national health insurance was the major obstacle against the use of bisphosphonates.


COVID-19 Restrictions and Adolescent Cigarette and E-cigarette Use in California.
Shelter-in-place orders altered facilitators and barriers to tobacco use (e.g., outlet closures, restricted social gatherings). This study examined whether the duration of time in shelter in place and compliance with different shelter-in-place orders influenced adolescent cigarette and E-cigarette use and how the use may differ by demographic characteristics. Shelter-in-place policy data obtained from government websites were merged with cross-sectional 2020 survey data on adolescents in California. Treatment variables included the proportion of time in shelter in place and self-reported compliance with shelter-in-place orders (for essential businesses and retail spaces and social and outdoor contexts). Multilevel logit models for dichotomous past 6-month cigarette and E-cigarette use and multilevel negative binomial regression models for past 6-month frequency of use were used. Moderation analyses were conducted on demographic measures. The sample included 1,196 adolescents (mean age=15.8 years, age range=13-19 years, 49.2% female, 50.0% White). Analyses were conducted in 2022. No associations were found between the proportion of time in shelter in place and outcomes. Shelter-in-place compliance with essential business and retail space orders was associated with lower odds of using cigarettes and E-cigarettes in the past 6 months. Compliance with social and outdoor context-related orders were associated with lower odds of using E-cigarettes and fewer days using cigarettes and E-cigarettes. Being aged ≥18 years moderated the associations between essential business/retail space and social/outdoor context-related shelter-in-place compliance orders and past 6-month frequency of cigarette smoking. Findings support tailored interventions for less compliant and older adolescents for future pandemic mitigation measures.


Cumulative risk analysis of carcinogenic contaminants in United States drinking water.
Cumulative risk analysis of contaminant occurrence in United States drinking water for the period of 2010-2017 indicates that over 100,000 lifetime cancer cases could be due to carcinogenic chemicals in tap water. The majority of this risk is due to the presence of arsenic, disinfection byproducts and radioactive contaminants. For different states within the U.S., cumulative cancer risk for drinking water contaminants ranges between 1 × 10-4 and 1 × 10-3, similar to the range of cumulative cancer risks reported for air pollutants. Overall, national attributable risk due to tap water contaminants is approximately 4 × 10-4, which is two orders of magnitude higher than the de minimus cancer risk of one-in-a-million. Thus, decreasing the levels of chemical contaminants in drinking water represents an important opportunity for protecting public health.


Management and Outcomes of Tibial Eminence Fractures in the Pediatric Population: A Systematic Review.
Tibial eminence fractures (TEF) of Meyers-McKeever type II-III-IV usually require surgical management. No consensus in the literature has been achieved regarding the best treatment option. The aims of the present systematic review were (1) to analyze the current literature and describe the outcomes of surgical treatment for TEF; and (2) to compare the outcomes of different surgical options using arthroscopic reduction and internal fixation (ARIF) with sutures or screws and open reduction and internal fixation (ORIF). A search was carried out with Pubmed, Medline, and Cochrane. Key terms were used "tibial" AND "eminence" or "spine" or "intercondylar" AND "paediatric" or "children" AND "fracture" or "avulsion" AND "treatment". Twelve articles met the inclusion criteria. Demographic data, clinical outcomes, and complication rates were evaluated for each study. Means/standard deviation and sum/percentage were used for continuous and categorical variables, respectively. Chi-square or t-student tests were applied. A p-value < 0.05 was considered statistically significant. ORIF showed superior clinical outcomes (Tegner (p < 0.05) and Lysholm (p < 0.001) scores) relative to ARIF and a lower incidence of arthrofibrosis (p < 0.05) and implant removal (p < 0.01). The Tegner, IKDC, and Lysholm scores showed statistically significant superior results following arthroscopic sutures compared to arthroscopic screws (p < 0.001). The incidence of arthrofibrosis was higher after arthroscopic sutures (p < 0.05), the implant removal was higher after screw fixation (p < 0.001) Conclusions: Better clinical results with low complication rates were achieved with ORIF surgery rather than ARIF; arthroscopic suture fixation resulted in higher clinical results compared to arthroscopic screw fixation and reduced the incidence of postoperative complications.


Effects of Foot Strike Techniques on Running Biomechanics: A Systematic Review and Meta-analysis.
Distance running is one of the most popular physical activities, and running-related injuries (RRIs) are also common. Foot strike patterns have been suggested to affect biomechanical variables related to RRI risks. To determine the effects of foot strike techniques on running biomechanics. The databases of Web of Science, PubMed, EMBASE, and EBSCO were searched from database inception through November 2018. The initial electronic search found 723 studies. Of these, 26 studies with a total of 472 participants were eligible for inclusion in this meta-analysis. Systematic review and meta-analysis. Level 4. Means, standard deviations, and sample sizes were extracted from the eligible studies, and the standard mean differences (SMDs) were obtained for biomechanical variables between forefoot strike (FFS) and rearfoot strike (RFS) groups using a random-effects model. FFS showed significantly smaller magnitude (SMD, -1.84; 95% CI, -2.29 to -1.38; P < 0.001) and loading rate (mean: SMD, -2.1; 95% CI, -3.18 to -1.01; P < 0.001; peak: SMD, -1.77; 95% CI, -2.21 to -1.33; P < 0.001) of impact force, ankle stiffness (SMD, -1.69; 95% CI, -2.46 to -0.92; P < 0.001), knee extension moment (SMD, -0.64; 95% CI, -0.98 to -0.3; P < 0.001), knee eccentric power (SMD, -2.03; 95% CI, -2.51 to -1.54; P < 0.001), knee negative work (SMD, -1.56; 95% CI, -2.11 to -1.00; P < 0.001), and patellofemoral joint stress (peak: SMD, -0.71; 95% CI, -1.28 to -0.14; P = 0.01; integral: SMD, -0.63; 95% CI, -1.11 to -0.15; P = 0.01) compared with RFS. However, FFS significantly increased ankle plantarflexion moment (SMD, 1.31; 95% CI, 0.66 to 1.96; P < 0.001), eccentric power (SMD, 1.63; 95% CI, 1.18 to 2.08;P < 0.001), negative work (SMD, 2.60; 95% CI, 1.02 to 4.18; P = 0.001), and axial contact force (SMD, 1.26; 95% CI, 0.93 to 1.6; P < 0.001) compared with RFS. Running with RFS imposed higher biomechanical loads on overall ground impact and knee and patellofemoral joints, whereas FFS imposed higher biomechanical loads on the ankle joint and Achilles tendon. The modification of strike techniques may affect the specific biomechanical loads experienced on relevant structures or tissues during running.


Structural properties and hydrolysability of recycled poplar residues (Populus L.): Effects of two-step acetic acid and sodium sulphite pre-treatment.
Poplar trees rapidly yield wood and are therefore suitable as a biofuel feedstock; however, the quality of poplar is modest, and the profitability of poplar cultivation depends on the efficiency of the harvesting process. This study offers a simple and sustainable technique to harvest lignocellulosic resources from poplar for bioethanol production. The proposed two-step pretreatment method increased the surface lignin content and decreased the surface polysaccharide content. The cellulose content increased to 54.9% and the xylan content decreased to 6.7% at 5% AC. The cellulose yield of poplar residues (Populus L.) reached 65.5% by this two-step acetic acid (AC) and sodium sulphite (SS) treatment method. Two-step pretreatment using 5% AC and 4% SS obtained a recovery of nearly 80% of the total available fermentable sugar. The surface characterization showed a higher porosity in treated samples, which improved their hydrolysability. This method decreased the amount of lignin in plant biomass, making it applicable for further wood resource recovery or waste recycling for biorefinery purposes at very low costs.


Mast cells and basophils are essential for allergies: mechanisms of allergic inflammation and a proposed procedure for diagnosis.
The current definition of allergy is a group of IgE-mediated diseases. However, a large portion of patients with clinical manifestations of allergies do not exhibit elevated serum levels of IgE (sIgEs). In this article, three key factors, ie soluble allergens, sIgEs and mast cells or basophils, representing the causative factors, messengers and primary effector cells in allergic inflammation, respectively, were discussed. Based on current knowledge on allergic diseases, we propose that allergic diseases are a group of diseases mediated through activated mast cells and/or basophils in sensitive individuals, and allergic diseases include four subgroups: (1) IgE dependent; (2) other immunoglobulin dependent; (3) non-immunoglobulin mediated; (4) mixture of the first three subgroups. According to our proposed definition, pseudo-allergic-reactions, in which mast cell or basophil activation is not mediated via IgE, or to a lesser extent via IgG or IgM, should be non-IgE-mediated allergic diseases. Specific allergen challenge tests (SACTs) are gold standard tests for diagnosing allergies in vivo, but risky. The identification of surface membrane activation markers of mast cells and basophils (CD203c, CCR3, CD63, etc) has led to development of the basophil activation test (BAT), an in vitro specific allergen challenge test (SACT). Based on currently available laboratory allergy tests, we here propose a laboratory examination procedure for allergy.


Glucocorticoids potentiate kainic acid-induced seizures and wet dog shakes.
Glucocorticoid effects on kainic acid-induced motor seizures and wet dog shakes in rats were investigated by adrenalectomy and dexamethasone treatment. One-day adrenalectomy attenuated kainic acid-induced wet dog shakes and seizure activity. These effects were restored by dexamethasone. Administration of dexamethasone to non-adrenalectomized rats potentiated kainic acid-induced wet dog shakes and severity of seizure activity. These results suggest that glucocorticoids may play an important role in modulating the severity of kainic acid-induced seizures and wet dog shakes.


Loss of integrated viral DNA sequences in polyomatransformed cells is associated with an active viral A function.
Rat cells transformed by polyoma virus contain, in addition to integrated viral DNA, a small number of nonintegrated viral DNA molecules. The free viral DNA originates from the integrated form through a spontaneous induction of viral DNA replication in a minority of the cell population. Its presence is under the control of the viral A locus. To determine whether the induction of free viral DNA replication was accompanied by a loss of integrated viral DNA molecules in a phenomenon similar to the "curing" of lysogenic bacteria, we selected for revertants arising in the transformed rat populations and determined whether these cells had lost integrated viral genomes. We further investigated whether the viral A function was necessary for "curing" by determining the frequency of cured cells in populations of rat cells transformed by the ts-a mutant of polyoma virus following propagation at the permissive or nonpermissive temperature. A large proportion of the revertants isolated were negative or weakly positive when assayed by immunofluorescence for polyoma T antigen and were unable to produce infectious virus upon fusion with permissive mouse cells. The T antigen-negative, virus rescue-negative clones can be retransformed by superinfection and appear to have lost a considerable proportion of integrated viral DNA sequences. Restriction enzyme analysis of the integrated viral DNA sequences shows that the parental transformed lines contain tandem repeats of integrated viral molecules, and that this tandem arrangement is generally lost in the cured derivatives. While cells transformed by wild-type virus undergo "curing" with about the same frequency at 33 degrees or 39 degrees C, cells transformed by the ts-a mutant contain a much higher frequency of cured cells after propagation at 33 degrees than at 39 degrees C. Our results indicate that in polyoma-transformed rat cells, loss of integrated viral DNA can occur at a rather high rate, producing (at least in some cases) cells which have reverted partially or completely to a normal phenotype. Loss of integrated viral DNA is never total and appears to involve an excision event. The polyoma A function (large T antigen) is necessary for such excision to occur. In the absence of a functional A gene product, the association of the viral DNA with the host DNA appears to be very stable.


Using human iPSC-derived kidney organoids to decipher SARS-CoV-2 pathology on single cell level.
We describe a protocol for single-cell RNA sequencing of SARS-CoV-2-infected human induced pluripotent stem cell (iPSC)-derived kidney organoids. After inoculation of kidney organoids with virus, we use mechanical and enzymatic disruption to obtain single cell suspensions. Next, we process the organoid-derived cells into sequencing-ready SARS-CoV-2-targeted libraries. Subsequent sequencing analysis reveals changes in kidney cells after virus infection. The protocol was designed for kidney organoids cultured in a 6-well transwell format but can be adapted to organoids with different organ backgrounds. For complete details on the use and execution of this protocol, please refer to Jansen et al. (2022).


Human mesenchymal stem cells treatment for severe COVID-19: 1-year follow-up results of a randomized, double-blind, placebo-controlled trial.
The long-term consequences of human umbilical cord-derived mesenchymal stem cell (UC-MSC) treatment for COVID-19 patients are yet to be reported. This study assessed the 1-year outcomes in patients with severe COVID-19, who were recruited in our previous UC-MSC clinical trial. In this prospective, longitudinal, cohort study, 100 patients enrolled in our phase 2 trial were prospectively followed up at 3-month intervals for 1 year to evaluate the long-term safety and effectiveness of UC-MSC treatment. The primary endpoint was an altered proportion of whole-lung lesion volumes measured by high-resolution CT. Other imaging outcomes, 6 min walking distance (6-MWD), lung function, plasma biomarkers, and adverse events were also recorded and analyzed. This trial was registered with ClinicalTrials.gov (NCT04288102). MSC administration improved in whole-lung lesion volume compared with the placebo with a difference of -10.8% (95% CI: -20.7%, -1.5%, p = 0.030) on day 10. MSC also reduced the proportion of solid component lesion volume compared with the placebo at each follow-up point. More interestingly, 17.9% (10/56) of patients in the MSC group had normal CT images at month 12, but none in the placebo group (p = 0.013). The incidence of symptoms was lower in the MSC group than in the placebo group at each follow-up time. Neutralizing antibodies were all positive, with a similar median inhibition rate (61.6% vs. 67.6%) in both groups at month 12. No difference in adverse events at the 1-year follow-up and tumor markers at month 12 were observed between the two groups. UC-MSC administration achieves a long-term benefit in the recovery of lung lesions and symptoms in COVID-19 patients. The National Key R&D Program of China, the Innovation Groups of the National Natural Science Foundation of China, and the National Science and Technology Major Project.


The anthropogenic fallout radionuclides in soils of Mount Khuko (the Western Caucasus) and their application for determination of sediment redistribution.
The purposes of this study are to determine the content and origin of anthropogenic fallout radionuclides (FRN) in soils of Mount Khuko, located in the western sector of the Caucasus Mountains and to assess the possibility to use them for evaluation of sediment redistribution for the alpine grasslands,. The field study was carried out in August 2019 near the top of Mount Khuko, located in the western part of the main Caucasus Mountain Ridge. Integral and incremental soil samples were collected from the different morphological units of the studied area. The content of 137Cs and 241Am in soil samples was evaluated using laboratory gamma-spectrometry. A part of samples was selected for Pu isotopes extraction and then alpha-spectrometric analysis. It was established that the 137Cs contamination of soils in the studied area has at least two sources of origin. The first source is the 137Cs bomb-derived fallout after the bomb tests in 1950-60th, which is widespread across the globe. The second source is 137Cs Chernobyl-derived fallout High random variability (Cv = 25-42%) was found within reference sites, located at the undisturbed areas on the local flat interfluves due to high variability of soil characteristics (grain size, density, organic matter content etc.). However minimum spatial variability (range 12,2-14,3 kBq/m2) was identified for the mean value of 137Cs inventories for all 5 reference sites located in the different parts of the studied area. It is difficult to separate individual peaks of the bomb-derived and Chernobyl-derived 137Cs falloutin sediment sinks with low sedimentation rates. Application 239,240Pu as an additional chronological marker allows to identify the origin of above mention peaks in the soils of alpine grasslands and of dry lake bottom.


Respiratory adaptation and surfactant composition of unanesthetized male and female lambs differ for up to 8 h after preterm birth.
Male preterm infants are more likely to experience respiratory distress syndrome than females. Our objectives were to determine if sex-related differences in physiological adaptation after preterm birth increase with time after birth and if the use of continuous positive airway pressure (CPAP) reduces these differences. Unanesthetized lambs (9F, 8M) were delivered at 0.90 of term. Blood gases, metabolites, and cardiovascular and respiratory parameters were monitored in spontaneously breathing lambs for 8 h. Supplemental oxygen was administered via a face mask at 4 cmH2O CPAP. At 8 h, lung compliance was determined, and bronchoalveolar lavage fluid (BALF) was analyzed for total protein and surfactant phospholipids. Surfactant protein (SP) gene expression and protein expression of SP-A and pro-SP-C were determined in lung tissue. For 8 h after delivery, males had significantly lower arterial pH and higher Paco2, and a greater percentage of males were dependent on supplemental oxygen than females. Inspiratory effort was greater and lung compliance was lower in male lambs. Total protein concentration in BALF, SP gene expression, and SP-A protein levels were not different between sexes; pro-SP-C was 24% lower in males. The use of CPAP did not eliminate the male disadvantage, which continues for up to 8 h after preterm birth.


[Glaucoma in the young patient. Clinical particularities].
Difficult to classify, glaucoma in young subjects--with a clinical presentation nevertheless comparable to the adult's--is underestimated, difficult to identify and treat, and is very heterogeneous. Few reliable screening tools are available and nothing has yet replaced the clinical experience of the physician. Vigilance is therefore required so as not to miss this diagnosis.


Ion transport blockers inhibit human rhinovirus 2 release.
Picornavirus replication causes leakage of cytoplasmic K+ and an influx of Na+ and Ca2+. In this study, we have explored the possibility that a blockade of Ca2+ and Na+ influx would reduce rhinovirus production and/or release. The Ca2+-channel blockers, verapamil and diltiazem, as well as the blocker of Na+/H+ exchange and the epithelial Na+ channel, EIPA, inhibited both virus production and release. The effect on virus release was more pronounced than the effect on production, thus raising the possibility that rhinovirus release may serve as a target for antiviral agents. Unexpectedly, our results also showed that the antiviral activity of the Ca2+-channel blockers was not due to the block of Ca2+ influx. Similarly, the antiviral activity of EIPA appeared to be unrelated to the blockade of cellular Na+/H+ exchanger or the epithelial Na+ channel. Potential alternative mechanisms of the antiviral activity of these compounds are discussed.


Peritoneal Flap for Lymphocele Prophylaxis Following Robotic-assisted Radical Prostatectomy with Lymph Node Dissection: The Randomised Controlled Phase 3 PELYCAN Trial.
Symptomatic lymphoceles (SLCs) after transperitoneal robotic-assisted radical prostatectomy with pelvic lymph node dissection (PLND) are common. Evidence from randomised controlled trials (RCTs) on the impact of peritoneal flaps (PFs) on lymphocele (LC) reduction is inconclusive. To show that addition of PFs leads to a reduction of postoperative SLCs. An investigator-initiated, prospective, parallel, double-blinded, adaptive, phase 3 RCT was conducted. Recruitment took place from September 2019 until December 2021; 6-month written survey-based follow-up was recorded. Stratification was carried out according to potential LC risk factors (extended PLND, diabetes mellitus, and anticoagulation) and surgeons; 1:1 block randomisation was used. Surgeons were informed about allocation after completion of the last surgical step. To create PFs, the ventral peritoneum was incised bilaterally and fixated to the pelvic floor. The primary endpoint was SLCs. Secondary endpoints included asymptomatic lymphoceles (ALCs), perioperative parameters, and postoperative complications. In total, 860 men were screened and 551 randomised. Significant reductions of SLCs (from 9.1% to 3.7%, p = 0.005) and ALCs (27.2% to 10.3%, p < 0.001) over the follow-up period of 6 mo were observed in the intention-to-treat analysis. Operating time was 11 min longer (p < 0.001) in the intervention group; no significant differences in amount (80 vs 103, p = 0.879) and severity (p = 0.182) of postoperative complications (excluding LCs) were observed. The survey-based follow-up might be a limitation. This is the largest RCT evaluating PF creation for LC prevention and met its primary endpoint, the reduction of SLCs. The results were consistent among all subgroup analyses including ALCs. Owing to the subsequent reduction of burden for patients and the healthcare system, establishing PFs should become the new standard of care. A new technique-creation of bilateral peritoneal flaps-was added to the standard procedure of robotic-assisted prostatectomy for lymph node removal. It was safe and decreased lymphocele development, a common postoperative complication and morbidity. Hence, it should become a standard procedure.


Comparison of stromal CD10 expression in benign, borderline, and malignant phyllodes tumors among Egyptian female patients.
Phyllodes tumors are group of biphasic fibroepithelial tumors of the breast of varying malignant potential, ranging from benign tumors to fully malignant sarcomas. According to the Egyptian National Cancer Institute, female malignant cases showed appreciable increase in the recent time period for breast cancer with the malignant phyllodes tumors representing 0.41% of cases in the year 2003-2004. This is an immunohistochemical study to compare CD10 expression in benign, borderline, and malignant phyllodes tumors, in order to highlight its diagnostic and prognostic values. This study conducted 34 Egyptian female cases of phyllodes tumors of different grades to be studied histologically and immunohistochemically using antibodies against CD10. The Chi-square test was used to determine differences in CD10 expression between benign, borderline, and malignant tumors. One-way ANOVA test was used to determine whether the difference was significant. Significance was established at P<0.05. In the 24 cases of benign phyllodes tumors, only four cases (16.7%) showed positive CD10 reactivity. Three cases (60%) out of five borderline phyllodes tumors showed positive CD10 reactivity, while four (80%) out of five cases of malignant phyllodes tumors showed positive CD10 staining. From these highly significant results, we believe that there is a strong correlation between CD10 expression and tumor grade, which could be an important observation that may have both diagnostic and prognostic implications as well as promising potential target for development of novel therapies.


Detection of thrombin using electrogenerated chemiluminescence based on Ru(bpy)3(2+)-doped silica nanoparticle aptasensor via target protein-induced strand displacement.
A sensitive and selective aptasensor using tri(2,2'-bipyridyl)ruthenium(II)-doped silica nanoparticles (Ru(bpy)3(2+)-doped SNPs) as DNA tags for detection of thrombin is developed based on the target protein-induced strand displacement of the DNA probe. For the proposed aptasensor, the aptamer was assembled on the surface of the Au electrode through Au-S binding. The hybridization event between the DNA probe labeled by the Ru(bpy)3(2+)-doped SNPs and the aptamer was evaluated by electrogenerated chemiluminescence (ECL) measurements. Then, the DNA probe was displaced by thrombin and the binding event between the thrombin and the aptamer was monitored by ECL measurements again. The difference of ECL intensity (deltaI(ECL)) of the two events could be used to quantify the thrombin. Other proteins, such as bovine serum albumin and bovine hemoglobin, had almost negligible deltaI(ECL). Under the optimal conditions, the deltaI(ECL) was linearly related to the concentration of the thrombin in the range of 10 fM to 10 pM and the detection limit was down to 1.0 fM since SNPs containing a large number of Ru(bpy)3(2+) molecules were labeled on the DNA probe.


Differential effects of fentanyl and morphine on intracellular Ca2+ transients and contraction in rat ventricular myocytes.
Our objective was to elucidate the direct effects of fentanyl and morphine on cardiac excitation-contraction coupling using individual, field-stimulated rat ventricular myocytes. Freshly isolated myocytes were loaded with fura-2 and field stimulated (0.3 Hz) at 28 degrees C. Amplitude and timing of intracellular Ca2+ concentration (at a 340:380 ratio) and myocyte shortening (video edge detection) were monitored simultaneously in individual cells. Real time Ca2+ uptake into isolated sarcoplasmic reticulum vesicles was measured using fura-2 free acid in the extravesicular compartment. The authors studied 120 cells from 30 rat hearts. Fentanyl (30-1,000 nM) caused dose-dependent decreases in peak intracellular Ca2+ concentration and shortening, whereas morphine (3-100 microM) decreased shortening without a concomitant decrease in the Ca2+ transient. Fentanyl prolonged the time to peak and to 50% recovery for shortening and the Ca2+ transient, whereas morphine only prolonged the timing parameters for shortening. Morphine (100 microM), but not fentanyl (1 microM), decreased the amount of Ca2+ released from intracellular stores in response to caffeine in intact cells, and it inhibited the rate of Ca2+ uptake in isolated sarcoplasmic reticulum vesicles. Fentanyl and morphine both caused a downward shift in the dose-response curve to extracellular Ca2+ for shortening, with no concomitant effect on the Ca2+ transient. Fentanyl and morphine directly depress cardiac excitation-contraction coupling at the cellular level. Fentanyl depresses myocardial contractility by decreasing the availability of intracellular Ca2+ and myofilament Ca2+ sensitivity. In contrast, morphine depresses myocardial contractility primarily by decreasing myofilament Ca2+ sensitivity.


New insights into the dynamics of sinusoidal endothelial fenestrae in liver sinusoidal endothelial cells.
Ultrastructural studies have shown that liver sinusoidal endothelial cells (LSECs) contain a cytoskeletal framework of filamentous actin, and that the presence of actin in the form of a calmodulin-actomyosin complex is responsible for regulation of the diameter of sinusoidal endothelial fenestrae (SEF). Rho has emerged as an important regulator of the actin cytoskeleton and consequently of cell morphology. We investigated actin filaments in relation to SEF in LSEC using heavy meromyosin decorated reaction and elucidated the roles of Rho and actin cytoskeleton in morphological and functional alterations of SEF. Second, according to intracytoplasmic Ca2+ concentration, plasma membrane Ca2+Mg2+-ATPase activities were clearly demonstrated on the outer surface of the labyrinth-like SEF in the isolated LSECs. Furthermore, by investigating intracytoplasmic Ca2+ concentration, we have demonstrated plasma membrane Ca2+Mg2+-ATPase activities on the outer surface of the labyrinth-like SEF in the isolated LSECs. Currently, the majority of fenestral studies are focused on finding ways to increase the liver sieve's porosity, which is reduced through pathological mechanisms.


Small GTP-binding proteins and their regulators in cardiac hypertrophy.
Small GTP-binding proteins (small G proteins) act as GDP-GTP-regulated molecular switches and are activated by guanine nucleotide exchange factors (GEFs) in response to diverse extracellular stimuli. During this last decade, numerous molecular and cellular studies, as well as genetically-modified animal models, have highlighted the role of small G proteins in the regulation of cardiac hypertrophy. The growing interest in small G protein signalling comes from the fact that chronic hypertrophic response is considered maladaptive and predisposes individuals to heart failure. Although some of the hypertrophic signalling pathways involving small G proteins have now been identified, a central question deals with the identity of the GEFs that modulate small G protein activation in the context of cardiac hypertrophy. Here, we discuss the precise regulation of Ras and Rho subfamilies of GTPases by GEFs and other regulatory proteins during cardiac hypertrophy. In addition, we summarize recent published data, mainly those describing the role of small G proteins in the development of myocardial hypertrophy and we further present the importance of their downstream effectors in myocardial remodelling.


Anatomical considerations of selective pudendal neurectomy.
The pudendal nerve was examined in 100 cadavers of both sexes. Because of the variable number of trunks of the pudendal nerve, 5 types were distinguished: one-trunked; two-trunked, rectopudendal; two-trunked, pudendo-penile (or -clitoridal); three-trunked, recto-perineo-penile (or -clitoridal); four-trunked, recto-perineo-perineopenile (or -clitoridal). The authors' suggestions make it possible to perform selective pudendectomy, consisting in denervation of the sphincter urethrae muscle by neurectomy of the perineal branches of the pudendal nerve.


Mechanisms of lead uptake and accumulation in wheat grains based on atmospheric deposition-soil sources.
Lead (Pb) accumulation in wheat grains depends on two aspects: i) Pb uptake by the roots and shoots, and ii) the translocation of organ Pb into the grain. However, the underlying mechanism of the uptake and transport of Pb in wheat remains unclear. This study explored this mechanism by establishing field leaf-cutting comparison treatments. Interestingly, as the organ with the highest Pb concentration, only 20.40 % of the root's relative contribution to grain Pb. The relative contributions of the spike, flag leaf, second leaf, and third leaf to grain Pb were 33.13 %, 23.57 %, 13.21 %, and 9.69 %, respectively, which was opposite to their Pb concentration distribution trends. According to Pb isotope analysis, it was found leaf-cutting treatments reduced the proportion of atmospheric Pb in grain, and grain Pb predominantly comes from atmospheric deposition (79.60 %). Furthermore, from the bottom to the top, the concentration of Pb in internodes decreased gradually, and the proportions of Pb originating from soil in the nodes also decreased, revealing that wheat nodes hindered the translocation of Pb from roots and leaves to the grain. Therefore, the hindering effect of nodes on the migration of soil Pb in wheat resulted in atmospheric Pb having a more convenient pathway to the grain than soil Pb, and further leading grain Pb accumulation primarily depended on the contribution of the flag leaf and spike.


Direct detection of intratumoral 5-fluorouracil trapping using metabolic 19F MR imaging.
The effective use of 5-fluorouracil (5-FU) in cancer therapy requires the noninvasive assessment of its transport, metabolism, and retention ("trapping") in the different tissues of the organism, particularly in the tumor. We used a chemical-shift selective 19F magnetic resonance (MR) imaging technique to map selectively 5-FU and its major catabolite alpha-fluoro-beta-alanine (FBAL) in six ACI rats bearing Morris hepatoma. After i.v. administration of 200 mg/kg-bw 5-FU, three metabolic MR maps were acquired consecutively in each animal: 1) an early 5-FU image (5-37 min post-injection (p.i.); dominant Fourier line, 8 min p.i.) characterizing the early uptake of 5-FU into the various tissues; 2) an FBAL image (40-72 min p.i.; dominant Fourier line, 56 min p.i.) reflecting the catabolism of the drug; and 3) a late 5-FU image (75-107 min p.i.; dominant Fourier line, 78 min p.i.) to assess the retention of unmetabolized 5-FU and its MR-visible anabolites. In the early 5-FU maps, the drug was detected in all major organs (e.g., heart, liver, kidneys) as well as in the muscular system. The FBAL maps showed no FBAL accumulation in the hepatoma which reveals that the tumor cells have lost hepatocellular functions relevant for 5-FU catabolism. On the late 5-FU maps, a significant amount of 5-FU was detected in only one of the six Morris hepatomas. The observation in this rat verifies directly that 5-FU can be trapped in solid tumors. The images, moreover, emphasize the necessity of acquiring spatially-resolved MR data to detect metabolic tumor heterogeneity.


Protein activator of cyclic AMP phosphodiesterase and cyclic nucleotide phosphodiesterase in bovine retina and bovine lens. Activity, subcellular distribution and kinetic parameters.
We have examined the activity of cyclic AMP phosphodiesterase, cyclic GMP phosphodiesterase and the protein activator of cyclic AMP phosphodiesterase in various anatomic and subcellular fractions of the bovine eye. Cyclic GMP hydrolysis was 1.6--12 times faster than hydrolysis of cyclic AMP in the subcellular fractions of the retina and in the precipitate of the rod outer segment. An opposite pattern was seen in the bovine lens, where the hyrolysis of cyclic AMP occurred 17 and 169 times faster than that of cyclic GMP in the supernatant and precipitate of lens, respectively. The activity of cyclic AMP phosphodiesterase was not affected by ethylene-glycol bis(beta-aminoethylether)-N,N'-tetraacetic acid in any fractions except in the retinal supernatant, suggesting that the phosphodiesterase exists primarily as a Ca2+-independent, activator-independent form. However, the protein activator of cyclic AMP phosphodiesterase existed in all fractions examine. A complex kinetic patternwas observed for both cyclic AMP and cyllic GMP hydrolysis by the 105000 times g lens supernatant. The Michaelis constants for both cyclic AMP (1.3-10(-6) and 9.I-10(-6) M) and cyclic GMP (1.04-10(6) AND 1.22 10(-5) M) appeared to be similar.


The residents' mental health status and community's role during the COVID-19 pandemic: a community-based cross-sectional study in China.
The related research of coronavirus disease 2019 (COVID-19) pandemic on the mental health of community residents is still lacking. Here we reported the mental health status of Chinese residents as well as the community's prevention and control during the pandemic period of COVID-19 and further explored the influencing factors of mental status. In this cross-sectional study, convenience sampling and snowball sampling methods were adopted from February 16 to February 23, 2020, and Chinese community residents were included according to the inclusion and exclusion criteria. Three questionnaires, including General Anxiety Disorder 7 (GAD-7), Patient Health Questionnaire 9 (PHQ-9), and a self-designed "Community prevention and control questionnaire", were used. A multivariate linear regression analysis was conducted to analyze the impact factors of anxiety and depression. A total of 3,001 community residents were included in this study. In total, 85.6% and 83.7% of participants had minimal anxiety and depression, respectively. Among all the community residents, 16.6% of participants proved that the communities they lived in had confirmed cases. 95.3% of participants reported that the residents were screened for mobility and contact history. 97.8% of participants reported the entrance and exit of the community were managed in their communities. There were 97.5% and 99% of participants needed to take body temperature and wear masks in their communities. Ninety-two percent (92%) of communities have their public areas and facilities disinfected daily, and 95.4% of communities have conducted health education about COVID-19. Factors including gender, education level, chronic illness, the frequency of going out, achieving information about COVID-19 by community and newspaper, and confirmed cases in the community, show association with community residents' anxiety and depression. The vast majority of Chinese residents have little anxiety and depression, and most communities have adopted standardized control measures following the government's regulations and policy, which plays a vital role in the control of COVID-19 and improving residents' anxiety and depression.


Ischemic damage increases nitric oxide production via inducible nitric oxide synthase in the cochlea.
The present study was designed to elucidate the dynamic changes of nitric oxide (NO) production in the perilymph and to investigate the immunostaining for inducible nitric oxide synthase (iNOS) in the cochlea for 7 days after transient cochlear ischemia. Moreover, aminoguanidine, which is a selective iNOS inhibitor, was administrated immediately following ischemia and every 24h thereafter for 7 days to investigate whether the production of NO is dependent on the iNOS pathway. Significant increases in the oxidative NO metabolites, nitrite (NO(2)(-)) and nitrate (NO(3)(-)), were measured on day 1 using an in vivo microdialysis and on-line high performance liquid chromatography (HPLC) system. The immunostaining for iNOS was strongly expressed on days 1 and 4 and returned to normal on day 7 after the ischemia. The administration of aminoguanidine reduced the oxidative NO metabolites on day 1 and suppressed the expression of iNOS. These findings suggest that transient ischemia causes a remarkable increase in NO production in the perilymph, which might be attributable to the iNOS pathway.


Primary lymphoma of the liver showing immunohistochemical evidence of T-cell origin. Successful management by right trisegmentectomy.
Primary malignant lymphomas of the liver are extremely rare. Thus far, 28 cases (1-13) of this entity have been described, but after careful clinical and histological staging, no other sites of extrahepatic involvement were demonstrated in 17 of these cases (2,3,5-13). We recently had the opportunity to investigate and manage a case of true primary malignant peripheral T-cell lymphoma of the liver in a 22-year-old man. To our knowledge this is the first reported case of a primary T-cell lymphoma of the liver. The patient was successfully treated with a right trisegmentectomy and no tumor recurrence has appeared after five years.


Activity profile of calpains I and II in chronically infarcted rat myocardium--influence of the calpain inhibitor CAL 9961.
1. The calpains have been proposed to be activated following cardiac ischaemia and to contribute to myocyte damage after myocardial infarction (MI). In this study, the activity of calpains I and II in the infarcted and non-infarcted rat myocardium and the action of the selective calpain inhibitor, CAL 9961, has been investigated. 2. MI was induced by permanent ligation of the left coronary artery. One, 3, 7 and 14 days post MI, the enzymes calpain I and II were separated from homogenates of the interventricular septum (IS) and left ventricular free wall (LVFW) by chromatography on DEAE-Sepharose. The activity of the calpains was measured in sham-operated and MI animals chronically treated with placebo or CAL 9961 (15 mg kg(-1) d(-1) s.c.) in a synthetic substrate assay. Treatment was started 3 days before MI induction. 3. Calpain I activity reached highest values in IS 14 days post MI, whereas maximum activity of calpain II was measured in LVFW 3 days post MI. In experiments in vitro, CAL 9961 completely inhibited both calpains. In vivo, chronic treatment of MI animals with CAL 9961 partially prevented the increase in calpain I activity in IS and reduced calpain II activity in LVFW to sham levels. 4. Our findings demonstrate that calpains I and II are activated after MI, however, both enzymes differ in their regional and temporal activation within the infarcted myocardium. Chronic inhibition of these enzymes with CAL 9961 might limit the calpain-induced myocardial damage and preserve cardiac structural integrity post MI.


Biomedical and health research: an analysis of country participation and research fields in the EU's Horizon 2020.
We analysed the Horizon 2020 project database, currently the European Union's (EU) largest framework programme for research and innovation-nearly 80 billion euros available over 7 years (2014-2020), to estimate the amount and type of EU-supported biomedical and health research and funding distribution among EU member states and non-European countries. Out of 20,877 projects as of 14th January 2019, a total of 4865 projects were classified as human health related. Ninety-four countries/territories worldwide participated in at least one biomedical project. The EU-15 original member states showed the highest participation as project leaders/partners and for acquired funding. Strong unequal funding distribution and participation between EU-15 and the 13 newest members-with EU-15 receiving about 87% of funding and EU-13 only 3%-have been evidenced. For both EU-15 and EU-13 we detected about 20% of projects involving the public and private sectors, according to Horizon 2020 guidelines. The largest percentage of projects was in the areas of biotechnological research (28.28%) and "basic research" (26.95%); these two sectors together accounted for 46.99% of the total funding assigned (7.9 billion euros). Research in neurosciences and neurological diseases appeared to be an increasing study area. Neurological and mental diseases covered about 21% of projects. Epidemiological studies accounted for about 5% of the total projects and for 14% of funding. Strong correlations were shown by indicators of financial and scientific capacity to identify success rates in obtaining EU funding, making the gap between countries with strong and weak research infrastructures difficult to overcome.


Forensic typing of short tandem repeat markers on the X and Y chromosomes.
Short tandem repeat (STR) markers are the cornerstone of forensic identity and kinship testing. Markers located on the X and the Y chromosome can complement those found on the autosomes, which are commonly used in laboratories today. The distinctive inheritance pattern of the sex chromosomes affords advantages to the investigation of family pedigrees required by mass disaster victim identification or missing persons cases, as well as mixed samples typically recovered in sexual assault crimes. This review aims to provide an overview of how X and Y STR markers are currently being used in forensic laboratories, including the applications which target their use, the markers and multiplexes facilitating recovery of this genetic information, and the tools available to interpret the resulting data. Emerging research topics are considered as well as issues requiring further study for both marker systems.


[The importance of the interference fringe visual acuity in the indication of a cataract-extraction (author's transl)].
100 patients have been examined before and after cataract-extraction using the Retinal Visual Acuity Tester, Takata, Japan. Laser produced interference fringes are formed on the retina, as a result the visual acuity can be determined easily before the operation. -- With immature cataracts we obtained a good correlation between the preoperative interference fringe visual acuity and the postoperative visual acuity tested by Landoltrings. Therefore, it can be differentiated whether worse visual acuity is caused by the cataract or by a retinal disease.


Antimicrobial susceptibility and beta-lactamase characterization of Branhamella catarrhalis isolates from 1983/1984 and 1988.
Branhamella catarrhalis isolates from lower respiratory tract specimens collected in 1983/84 (n = 50) and 1988 (n = 30) were examined for beta-lactamase production. The percent of beta-lactamase-producing strains increased from 25% to 63% from 1983/84 to 1988. beta-lactamases from 30 strains could be typed, and of these 28 were of the BRO-1 type and two the BRO-2 type. The two beta-lactamase inhibitors, clavulanic acid and brobactam were very active against beta-lactamase extracts whether of the BRO-1 or the BRO-2 type. Susceptibility to a number of antimicrobial agents or combinations of agents was determined by a plate dilution method (MICs) and by a tablet diffusion method. The penicillin tablet (Rosco Neo-Sensitabs) was found to be useful in discriminating between beta-lactamase-producing and non-beta-lactamase- producing strains of B. catarrhalis.


[Surgical treatment of recurrent dislocation of the patella in children].
To study an operative method for repair of recurrent dislocation of the patella in children. 36 children with recurrent patellar dislocation were treated surgically at an average age of 9.1 years. They were followed on average for 4 years and 4 months. Nine of the 36 children underwent bilateral repair, with 45 knees were treated. The combined soft tissue operations included lateral retinacular releasing, medial retinacular tightening of the knee, vastus medialis muscle transfer to the patella, medial and distal transfer of the half patellar tendon. 28 patients had stable knee joint and a full range of motion, and improved sports activity after operation. Seven children achieved satisfactory function of the knee joint but there was no significant difference between pre-and postoperative sports exercise ability. One child suffered from a redislocation of the patella. Wound infection was not seen and the range of knee joint motion was not restricted in all patients. The combined procedures for repair of recurrent dislocation of the patella in children risk free for growth disturbances secondary to iatrogenic injury to the proximal tibial physis. It is safe and effective for recurrent dislocation of the patella in skeletally immature children.


Interrelation of the stagnant slab, Ontong Java Plateau, and intraplate volcanism as inferred from seismic tomography.
We investigated the seismological structure beneath the equatorial Melanesian region, where is tectonically unique because an immense oceanic plateau, a volcanic chain and subduction zones meet. We conducted a multi-frequency P-wave tomography using data collected from an approximately 2-year-long seismic experiment around the Ontong Java Plateau (OJP). High-velocity anomalies were revealed beneath the center of the OJP at a depth of ~ 150 km, the middle-eastern edge of the OJP at depths of 200-300 km, and in the mantle transition zone beneath and around the OJP; low-velocity anomalies were observed along the Caroline volcanic island chain above 450 km depth. These anomalies are considered to be associated with the thick lithosphere of the OJP, remnant dipping Pacific slab, stagnant Pacific slab, and a sheet-like upwelling. The broad stagnant slab was formed due to rapid trench retreat from 48 to 25 Ma until when the OJP with thick lithosphere collided with a subduction boundary of the Pacific and Australian plates. This collision triggered slab breakoff beneath the arc where the dipping slab remained. The stagnant Pacific slab in the mantle transition zone restricted the plume upwelling from the lower mantle causing sheet-like deformed upwelling in the upper mantle.


Large intracoronary thrombus and its management during primary PCI.
Large intracoronary thrombus has been reported in significant number of patients with STEMI. Primary PCI is the current standard of care in patients of STEMI. Despite the availability of dual antiplatelets, GP IIb/IIIa inhibitor and effective anticoagulation regimens, large intracoronary thrombus remains one of the biggest challenge to interventional cardiologists during primary PCI. Large intracoronary thrombus may lead to distal embolization, no/slow reflow or embolization into a non-culprit vessel and is associated with adverse cardiovascular outcome. There is no ideal management strategy. We hereby discuss the current available methods/strategies to deal with large thrombus burden encountered during primary PCI, in the current manuscript.


Effects of intravenously administered tetra-thiomolybdate on the distribution of copper in the liver and kidney of copper loaded sheep: a histochemical study.
The effects of intravenously administered thiomolybdate on the liver and kidney of copper loaded sheep were studied using 16 ewes in three groups. Copper, iron and molybdenum concentrations were determined by spectrophotometry and the distribution of copper in the liver and kidney was studied histochemically. Following thiomolybdate administration, the concentration of copper in the liver was reduced, that of molybdenum increased and the concentration of copper and molybdenum in the kidney increased. The reduction of copper concentration in the liver was associated with reductions in the number and size of granules in hepatocytes which stained positively for copper and in the number of Kupffer cells containing positively staining granules. The decrease in the amount of copper in hepatocytes appeared to be greater than that in Kupffer cells. This effect was greatest in the centrilobular zones and least in the periportal zones. The increased concentration of copper and molybdenum in kidney was associated with an increase in the number and size of granules staining positively for copper in the epithelial cells of the proximal convoluted tubules which suggested an uptake of copper-molybdenum complexes by the lysosomes of these cells.


Neurons can upregulate Cav-1 to increase intake of endothelial cells-derived extracellular vesicles that attenuate apoptosis via miR-1290.
Extracellular vesicles (EVs) including exosomes can serve as mediators of cell-cell communication under physiological and pathological conditions. However, cargo molecules carried by EVs to exert their functions, as well as mechanisms for their regulated release and intake, have been poorly understood. In this study, we examined the effects of endothelial cells-derived EVs on neurons suffering from oxygen-glucose deprivation (OGD), which mimics neuronal ischemia-reperfusion injury in human diseases. In a human umbilical endothelial cell (HUVEC)-neuron coculture assay, we found that HUVECs reduced apoptosis of neurons under OGD, and this effect was compromised by GW4869, a blocker of exosome release. Purified EVs could be internalized by neurons and alleviate neuronal apoptosis under OGD. A miRNA, miR-1290, was highly enriched in HUVECs-derived EVs and was responsible for EV-mediated neuronal protection under OGD. Interestingly, we found that OGD enhanced intake of EVs by neurons cultured in vitro. We examined the expression of several potential receptors for EV intake and found that caveolin-1 (Cav-1) was upregulated in OGD-treated neurons and mice suffering from middle cerebral artery occlusion (MCAO). Knock-down of Cav-1 in neurons reduced EV intake, and canceled EV-mediated neuronal protection under OGD. HUVEC-derived EVs alleviated MCAO-induced neuronal apoptosis in vivo. These findings suggested that ischemia likely upregulates Cav-1 expression in neurons to increase EV intake, which protects neurons by attenuating apoptosis via miR-1290.


Ethnic differences in ambient air pollution and risk of acute ischemic stroke.
To investigate the association between short-term changes in ambient pollution (particulate matter <2.5 μm in aerodynamic diameter (PM2.5) and ozone (O3)) and the risk of ischemic stroke among individuals living in a bi-ethnic community and whether this association is modified by ethnicity. We identified incident ischemic stroke cases from the population-based Brain Attack Surveillance in Corpus Christi (BASIC) project between 2000 and 2012. Associations between PM2.5 (mean 24-h) and O3 (maximal 8-h) levels, measured on the same-day and lags of 1-3 days, and odds of ischemic stroke were assessed using a time-stratified case-crossover design and modeled using conditional logistic regression. We explored race/ethnicity (Mexican American versus non-Hispanic white) as a modifier by including interaction terms in the models. There were 2948 ischemic strokes with median age 71 years (IQR: 59-80). We observed no overall associations between the air pollutants and odds of ischemic stroke at any lag. When stratified by ethnicity, higher O3 was consistently associated with greater odds of ischemic stroke for non-Hispanic whites, but not for Mexican Americans. Higher PM2.5 was generally associated with lower odds of ischemic stroke for non-Hispanic whites but modestly greater odds for Mexican Americans. Ethnic differences in the associations between ischemic stroke and short-term exposures to O3 and PM2.5 were suggested indicating that further study in diverse populations may be warranted.


[The interaction of mu-opioid receptors with intracellular signal systems].
Review is devoted a modem state of researches of micro-opioid receptor. It is discussed literature data on the integration of micro-opioid receptor with adenylyl cyclase, phospholipase A2, C and D. It is analyzed reports on the interaction of micro-opioid receptor with NO-synthase and guanylyl cyclase.


Radiotherapy enhanced by cis-platinum in stage III non-small cell lung cancer: a phase II study.
From January 1984 to December 1986, 94 patients with unresectable, locally advanced, non-small cell lung cancer (NSCLC) were treated to assess both the efficacy and the toxicity of a combined modality treatment including radiation therapy (45 Gy/15 fractions/3 weeks) and daily low dose cDDP (6 mg/m2). The overall response rate for the 90 evaluable patients was 54.3% with 16.6% of complete responses. At a minimum follow-up of 4 years, the overall median survival time was 12 months. Provided adequate hydration is ensured, the cDDP regimen chosen as a radiosensitizer can be safely combined with radiation therapy.


Self-assembly formed by a short DNA probe pair: Application for highly sensitive mRNA species detection without reverse transcription.
We describe a novel technology for detecting nucleic acids: Probe Alteration Link Self-Assembly Reactions (PALSAR). PALSAR comprises DNA self-assembly of pairs of short DNA probes formed by alternate hybridization of three complementary regions in a pair of honeycomb probes (HCPs). Self-assembly occurs at designated salt concentrations and reaction temperatures and requires no enzymes. We prepared pairs of HCPs to detect mRNAs encoded by the GAPDH gene β-actin (BA) gene, CD3D gene, CD4 gene, major vault protein (MV) gene and the signalling lymphocytic activation molecule-associated protein (SAP) gene, and succeeded in quantitatively detecting these mRNAs. PALSAR could detect mRNA directly without synthesizing cDNA. Moreover, multiple mRNAs could be detected simultaneously in a single reaction tube and there was a good correlation between the results obtained PALSAR and those by real-time PCR.


Phase I study of CBM.CD19 chimeric antigen receptor T cell in the treatment of refractory diffuse large B-cell lymphoma in Chinese patients.
Anti-CD19 chimeric antigen receptor (CAR) T cell therapy has shown impressive efficacy in treating B-cell malignancies. A single-center phase I dose-escalation study was conducted to evaluate the safety and efficacy of T cells transduced with CBM.CD19 CAR, a second-generation anti-CD19 CAR bearing 4-1BB costimulatory molecule, for the treatment of patients with refractory diffuse large B-cell lymphoma (DLBCL). Ten heavily treated patients with refractory DLBCL were given CBM.CD19 CAR-T cell (C-CAR011) treatment. The overall response rate was 20% and 50% at 4 and 12 weeks after the infusion of C-CAR011, respectively, and the disease control rate was 60% at 12 weeks after infusion. Treatment-emergent adverse events occurred in all patients. The incidence of cytokine release syndrome in all grades and grade ⩾ 3 was 90% and 0, respectively, which is consistent with the safety profile of axicabtagene ciloleucel and tisagenlecleucel. Neurotoxicity or other dose-limiting toxicities was not observed in any dose cohort of C-CAR011 therapy. Antitumor efficacy was apparent across dose cohorts. Therefore, C-CAR011 is a safe and effective therapeutic option for Chinese patients with refractory DLBCL, and further large-scale clinical trials are warranted.


Association of maternal chronic arsenic exposure with the risk of neural tube defects in Northern China.
Numerous animal studies have shown that high arsenic exposure can induce neural tube defect (NTD) formation. However, epidemiological evidence related to this finding is scarce. The key objective of our study is to evaluate whether maternal arsenic exposure is associated with NTD risk in Northern China. Our case-control study was conducted in 11 countries or cities in Shanxi and Hebei provinces during 2003-2007. A total of 774 mothers were included as participants: 511 controls and 263 cases (including 123 with anencephaly, 115 with spina bifida, 18 with encephalocele, and 7 with other NTD subtypes). The arsenic concentration was measured in a specific section of hair that grew from 3 months before to 3 months after conception. We found a higher hair arsenic concentration in the NTD cases with median (inter-quartile range) of 0.093 (0.025-0.387) μg/g hair than that in the controls with a value of 0.082 (0.030-0.414) μg/g hair. Maternal hair arsenic concentration above its median of the controls was associated with an increased risk of the total NTDs with an adjusted odds ratio (OR) of 1.32 [95% confidence interval (CI): (0.91-1.92)], which was not statistically significant (p = 0.14), although the crude OR without adjusting for the confounders of 1.68 (95% CI: 1.24-2.27; p < 0.001) suggested that hair arsenic is a risk factor of NTDs. There was no dose-response relationship between maternal hair arsenic concentration and the risk of total NTDs. Similar phenomena were found for anencephaly and spina bifida, respectively. Overall, our findings showed that maternal periconceptional arsenic exposure may not significantly contribute to the risk of NTD development in Northern China; other risk factors need to be further examined in future studies.


Large Carrier Mobilities in ErMnO3 Conducting Domain Walls Revealed by Quantitative Hall-Effect Measurements.
Kelvin probe force microscopy (KPFM) has been used to directly and quantitatively measure Hall voltages, developed at conducting tail-to-tail domain walls in ErMnO3 single crystals, when current is driven in the presence of an approximately perpendicular magnetic field. Measurements across a number of walls, taken using two different atomic force microscope platforms, consistently suggest that the active p-type carriers have unusually large room temperature mobilities of the order of hundreds of square centimeters per volt second. Associated carrier densities were estimated to be of the order of 1013 cm-3. Such mobilities, at room temperature, are high in comparison with both bulk oxide conductors and LaAlO3-SrTiO3 sheet conductors. High carrier mobilities are encouraging for the future of domain-wall nanoelectronics and, significantly, also suggest the feasibility of meaningful investigations into dimensional confinement effects in these novel domain-wall systems.


Correction: Early Behavioral Abnormalities and Perinatal Alterations of PTEN/AKT Pathway in Valproic Acid Autism Model Mice.
[This corrects the article DOI: 10.1371/journal.pone.0153298.].


Intensive Care Unit-Acquired Weakness.
Intensive care unit-acquired weakness (ICUAW) is a substantial contributor to long-term disability in survivors of critical illness. Critical illness polyneuropathy, critical illness myopathy, and muscle atrophy from disuse contribute in various proportions to ICUAW. ICUAW is a clinical diagnosis supported by electrophysiology and newer diagnostic tests, such as muscle ultrasound. Risk factor reduction, including the aggressive treatment of sepsis and early mobilization, improves outcome. Although some patients with ICUAW experience a full recovery, for others improvement is slow and incomplete and quality of life is adversely affected. This article examines aspects of ICUAW and identifies potential areas of further study.


Response-reinforcement relationships in chronic pain syndrome: applicability of Herrnstein's law.
The quantitative relationship between chronic pain behavior and reinforcement has been generally found to be positive. Herrnstein's statement of the law of effect adds a qualification: it predicts that response rate varies hyperbolically with contingent reinforcement, and that response rate is also negatively related to extraneous reinforcement. The present study is an original investigation of the first of these assertions with regard to the naturally occurring behaviors in chronic pain syndrome. It was found that the stereotypic pain behaviors of 12 chronic pain patients were hyperbolically rather than proportionally related to contingent reinforcement from their significant others. Likewise, healthy behaviors in the same group were a hyperbolic function of contingent reinforcement. Estimates of extraneous reinforcement were also obtained for each category of behaviors. As a group, the subjects showed an inverse relationship between pain behaviors and healthy behaviors, consistent with previous research in the pain area. The present findings add generality to Herrnstein's matching law which serves as a descriptive model of target behaviors in chronic pain patients. The findings also have prescriptive implications for the modification of behavior among chronic pain sufferers.


Fungal Nails? DNA Facts Challenge Dystrophic Etiology.
Historically recalcitrant to treatment, infection of the nail unit is a pervasive clinical condition affecting approximately 10% to 20% of the US population; patients present with both cosmetic symptomatology and pain, with subsequent dystrophic morphology. To date, the presumptive infectious etiologies include classically reported fungal dermatophytes, nondermatophyte molds, and yeasts. Until now, the prevalence and potential contribution of bacteria to the clinical course of dystrophic nails had been relatively overlooked, if not dismissed. Previously, diagnosis had largely been made by means of clinical presentation, although microscopic examinations (potassium hydroxide) of nail scrapings to identify fungal agents and, more recently, panel-specific polymerase chain reaction assays have been used to elucidate causative infectious agents. Each of these tools suffers from test-specific limitations. Molecular-age medicine now includes DNA-based tools to universally assess any microbe or pathogen with a known DNA sequence. This affords clinicians with rapid DNA sequencing technologies at their disposal. These sequencing-based diagnostic tools confer the accuracy of DNA-level certainty, and concurrently obviate cultivation or microbial phenotypical biases. Using DNA sequencing-based diagnostics, the results in this article document the first identification and quantification of significant bacterial, rather than mycotic, pathogens to the clinical manifestation of dystrophic nails. In direct opposition to the prevailing and presumptive mycotic-based causes, the results in this article invoke questions about the very basis for our current standards of care, including effective treatment regimens.


A simple, high-throughput stabilization assay to test HIV-1 uncoating inhibitors.
Shortly after entering the cell, HIV-1 copies its genomic RNA into double-stranded DNA in a process known as reverse transcription. This process starts inside a core consisting of an enclosed lattice of capsid proteins that protect the viral RNA from cytosolic sensors and degradation pathways. To accomplish reverse transcription and integrate cDNA into the host cell genome, the capsid shell needs to be disassembled, or uncoated. Premature or delayed uncoating attenuates reverse transcription and blocks HIV-1 infectivity. Small molecules that bind to the capsid lattice of the HIV-1 core and either destabilize or stabilize its structure could thus function as effective HIV-1 inhibitors. To screen for such compounds, we modified our recently developed FAITH assay to allow direct assessment of the stability of in vitro preassembled HIV-1 capsid-nucleocapsid (CANC) tubular particles. This new assay is a high-throughput fluorescence method based on measuring the amount of nucleic acid released from CANC complexes under disassembly conditions. The amount of disassembled CANC particles and released nucleic acid is proportional to the fluorescence signal, from which the relative percentage of CANC stability can be calculated. We consider our assay a potentially powerful tool for in vitro screening for compounds that alter HIV disassembly.


The genetic relationship of SOX9 polymorphisms with osteoarthritis risk in Chinese population: A case-control study.
This research aimed to reveal the relationship of SRY-type HMG box 9 (SOX9) gene polymorphisms with osteoarthritis (OA) risk in a Chinese population.Polymerase chain reaction and direct sequencing were used for genotyping polymorphism in 152 OA patients and 139 controls. Firstly, the conformity of genotype distribution to Hardy-Weinberg equilibrium in the control group was checked. The differences in genotype and allele frequencies of our studied polymorphism were compared between the two groups using chi-square test. Odds ratio (OR) with 95% confidence interval (95%CI) was used to appraise the strength of the relationship between the polymorphism and OA occurrence. Cross-over analysis was conducted to reveal the interaction between polymorphisms in SOX9.The AA genotype of the polymorphism rs1042667 was significantly correlated to the increased susceptibility to OA (OR = 2.075, 95%CI = 1.042-4.132). We also detected that the A allele of the polymorphism rs1042667 also obviously increased the occurrence of OA in our study (OR = 1.401, 95%CI = 1.009-1.945). Moreover, the G allele of the polymorphism rs12601701 and the A allele of the polymorphism rs1042667 could significantly elevate the risk of OA (OR = 2.075, 95%CI = 1.021-4.218).SOX9 polymorphism rs1042667 may be a risk factor for OA in Chinese Han population. The interaction between the polymorphisms rs1042667 and rs12601701 also contribute to OA risk.


Safety and efficacy assessment of aerogels for biomedical applications.
The unique physicochemical properties of aerogels have made them an attractive class of materials for biomedical applications such as drug delivery, regenerative medicine, and wound healing. Their low density, high porosity, and ability to regulate the pore structure makes aerogels ideal nano/micro-structures for loading of drugs and active biomolecules. As a result of this, the number of in vitro and in vivo studies on the therapeutic efficacy of these porous materials has increased substantially in recent years and continues to be an area of great interest. However, data about their in vivo performance and safety is limited. Studies have shown that polymer-based, silica-based and some hybrid aerogels are generally regarded as safe but given that studies on the acute, subacute, and chronic toxicity for the majority of aerogel types is missing, more work is still needed. This review presents a comprehensive summary of different biomedical applications of aerogels proposed to date as well as new and innovative applications of aerogels in other areas such as decontamination. We have also reviewed their biological effect on cells and living organisms with a focus on therapeutic efficacy and overall safety (in vivo and in vitro).


Living in honey: bacterial and fungal communities in honey of sympatric populations of Apis mellifera and the stingless bee Melipona beecheii, in Yucatan, Mexico.
Bacterial and fungal communities in the honey of sympatric populations of the bee species Apis mellifera and Melipona beecheii were profiled by amplicon sequencing of the 16S gene and the ITS of the ribosomal DNA. Results showed that the structure of the honey microbiota of these two bee species was very different from each other. Both the bacterial and fungal species in A. mellifera honey were more similar to those of A. mellifera honey reported for other parts of the world than to those in M. beecheii honey. Nevertheless, in both, the most abundant bacterial species belonged to the family Lactobacillaeae.


A PB1 T296R substitution enhance polymerase activity and confer a virulent phenotype to a 2009 pandemic H1N1 influenza virus in mice.
While the 2009 pandemic H1N1 virus has become established in the human population as a seasonal influenza virus, continued adaptation may alter viral virulence. Here, we passaged a 2009 pandemic H1N1 virus (A/Changchun/01/2009) in mice. Serial passage in mice generated viral variants with increased virulence. Adapted variants displayed enhanced replication kinetics in vitro and vivo. Analysis of the variants genomes revealed 6 amino acid changes in the PB1 (T296R), PA (I94V), HA (H3 numbering; N159D, D225G, and R226Q), and NP (D375N). Using reverse genetics, we found that a PB1-T296R substitution found in all adapted viral variants enhanced viral replication kinetics in vitro and vivo, increased viral polymerase activity in human cells, and was sufficient for enhanced virulence of the 2009 pandemic H1N1 virus in mice. Therefore, we defined a novel influenza pathogenic determinant, providing further insights into the pathogenesis of influenza viruses in mammals.


Deuterium NMR in the solid-state and in solution of the molecular motion of the bases in poly(I) and poly(I) . poly(C).
To provide information regarding the conformational flexibility of nucleic acids, in particular the rate and amplitude of base motions, we have observed the deuterium NMR from single-stranded and double-stranded polynucleotides. Poly(I) was deuterated at the 8-position of the base, and the deuterium NMR was examined in solution (at 23.0 and 55.4 MHz) and for hydrated and dry fibers (at 23.0 MHz). In the solid state, the deuterium signal of dry poly(I) exhibits a powder pattern with the maximal expected quadrupolar splitting, while the relatively short spin-lattice relaxation time indicates the presence of a rapid internal reorientation of the C-D bond with an amplitude of that motion of at least +/- 2.4 degrees. Hydrating the poly(I) fibers to the extent of eight molecules of water per nucleotide results in the disappearance of the deuterium signal, apparently due to a decreased spin-spin relaxation time shorter than the instrumental dead-time (even using the quadrupolar echo technique); this could occur if conformational fluctuations are occurring at a rate comparable to the deuterium quadrupole interaction strength, i.e., 175 kHz. In solution, a theoretical fit to the measured Lorentzian linewidths and spin-lattice relaxation times necessitates the inclusion of at least two motional correlation times, with a subnanosecond internal motion. Double-stranded poly(I) . poly(C) yielded a solid state spectrum similar to poly(I), albeit with a longer T1, which reduced the lower limit for the amplitude of an internal motion to +/- 1.9 degrees. The 2H signal from the poly(I) . poly(C), hydrated to a degree of approx. eight molecules of water per base pair, retained its solid-state lineshape (with a reduced T1 value, indicating increased internal mobility of the bases with a lower limit on amplitude of +/- 4.7 degrees). In solution, however, the 2H-NMR signal from poly(I) . poly(C) became virtually undetectable, even in solid-echo experiments, when the echo was observed after 52 microseconds. This indicates that the spin-spin relaxation time of the deuterium nucleus must be close to its theoretical minimum of about 9 microseconds, and the correlation time for an isotropic reorientation of the C-D vector can be estimated to be between 0.2 and 200 microseconds.


Percutaneous Drainage of Chronic Destructive Lumbar Osteomyelitis Abscess Via the Use of Bilateral Transpedicular Trocar Access.
Pyogenic spondylodiskitis is an infection of intervertebral disks and spinal vertebral bodies. Various minimally invasive approaches to the infected disk spaces/abscesses have been described for management of early stages of the infection. Patients with chronic occurrence present with extensive infection, neurologic deficits, and bone destruction. Such patients commonly have substantial medical comorbidities. Despite the increased risks of complications, they often are treated with open surgical approaches without minimally invasive options. We describe a bilateral transpedicular approach to vertebral body abscess in a chronically infected patient with intraoperative contiguous irrigation. We present 2 cases, a 58-year-old man and a 61-year-old man, both with a diagnosis of vertebral osteomyelitis. Images of lumbar spine showed epidural abscess and adjacent vertebral body destruction. Because of their poor clinical condition and chronicity of disease, these patients underwent percutaneous bilateral transpedicular approach. Patients in poor health and with chronic vertebral osteomyelitis may benefit from minimally invasive percutaneous transpedicular drainage and irrigation of the abscess, representing a minimally invasive and effective treatment alternative for these patients.


Development and evaluation of machine learning models based on X-ray radiomics for the classification and differentiation of malignant and benign bone tumors.
To develop and validate machine learning models to distinguish between benign and malignant bone lesions and compare the performance to radiologists. In 880 patients (age 33.1 ± 19.4 years, 395 women) diagnosed with malignant (n = 213, 24.2%) or benign (n = 667, 75.8%) primary bone tumors, preoperative radiographs were obtained, and the diagnosis was established using histopathology. Data was split 70%/15%/15% for training, validation, and internal testing. Additionally, 96 patients from another institution were obtained for external testing. Machine learning models were developed and validated using radiomic features and demographic information. The performance of each model was evaluated on the test sets for accuracy, area under the curve (AUC) from receiver operating characteristics, sensitivity, and specificity. For comparison, the external test set was evaluated by two radiology residents and two radiologists who specialized in musculoskeletal tumor imaging. The best machine learning model was based on an artificial neural network (ANN) combining both radiomic and demographic information achieving 80% and 75% accuracy at 75% and 90% sensitivity with 0.79 and 0.90 AUC on the internal and external test set, respectively. In comparison, the radiology residents achieved 71% and 65% accuracy at 61% and 35% sensitivity while the radiologists specialized in musculoskeletal tumor imaging achieved an 84% and 83% accuracy at 90% and 81% sensitivity, respectively. An ANN combining radiomic features and demographic information showed the best performance in distinguishing between benign and malignant bone lesions. The model showed lower accuracy compared to specialized radiologists, while accuracy was higher or similar compared to residents. • The developed machine learning model could differentiate benign from malignant bone tumors using radiography with an AUC of 0.90 on the external test set. • Machine learning models that used radiomic features or demographic information alone performed worse than those that used both radiomic features and demographic information as input, highlighting the importance of building comprehensive machine learning models. • An artificial neural network that combined both radiomic and demographic information achieved the best performance and its performance was compared to radiology readers on an external test set.


Fabrication and in vitro evaluation of a sponge-like bioactive-glass/gelatin composite scaffold for bone tissue engineering.
In this work a bioactive composite scaffold, comprised of bioactive-glass and gelatin, is introduced. Through direct foaming a sponge-like composite of a sol-gel derived bioactive-glass (70S30C; 70% SiO2, 30% CaO) and porcine gelatin was developed for use as a biodegradable scaffold for bone tissue engineering. The composite was developed to provide a suitable alternative to synthetic polymer based scaffolds, allowing directed regeneration of bone tissue. The fabricated scaffold was characterised through X-ray microtomography, scanning electron and light microscopy demonstrating a three dimensionally porous and interconnected structure, with an average pore size (170 μm) suitable for successful cell proliferation and tissue ingrowth. Acellular bioactivity was assessed through apatite formation during submersion in simulated body fluid (SBF) whereby the rate and onset of apatite nucleation was found to be comparable to that of bioactive-glass. Modification of dehydrothermal treatment parameters induced varying degrees of crosslinking, allowing the degradation of the composite to be tailored to suit specific applications and establishing its potential for a wide range of applications. Use of genipin to supplement crosslinking by dehydrothermal treatment provided further means of modifying degradability. Biocompatibility of the composite was qualified through successful cultures of human dental pulp stem cells (HDPSCs) on samples of the composite scaffold. Osteogenic differentiation of HDPSCs and extracellular matrix deposition were confirmed through positive alkaline phosphatase staining and immunohistochemistry.


Velocity field distribution control in antisolvent flow realizing highly stable and efficient perovskite nanocrystals.
Achieving highly stable and efficient perovskite nanocrystals (NCs) without applying functional additives or encapsulation, particularly sustaining the stability in ultra-dilute solution, is still a formidable challenge. Here, we show the FAPbI3 perovskite NCs with achieved ∼100 % photoluminescence quantum yield (PLQY) and low defect density (∼0.2 cm-3 per NCs), which is obtained by controlling the velocity field distribution of antisolvent flow in ligand-assisted reprecipitation process. The NCs show incredible reproducibility with narrow deviation of PLQY and linewidth between batch by batch, as well as remarkable stability of maintaining over 80 % PLQY, either in an ultra-diluted solution (9.3 × 10-6 mg/mL), or storing in ambient condition after 90 days with concentration of 0.09 mg/mL. The results in this work demonstrate the interplay of fluid mechanics and crystallization kinetics of perovskite, which pioneers a novel and unprecedent understanding for improving the stability of perovskite NCs for efficient quantum light source.


Molecular cloning of chicken myosin-binding protein (MyBP) H (86-kDa protein) reveals extensive homology with MyBP-C (C-protein) with conserved immunoglobulin C2 and fibronectin type III motifs.
The complete nucleotide sequence of a cDNA clone encoding the chicken skeletal muscle myosin-binding protein H (MyBP-H), formerly termed 86-kDa protein, has been established and the predicted amino acid sequence compared with other proteins entered into the GenBank data base. The full-length cDNA of 2066 base pairs contains a single open reading frame of 1611 base pairs encoding a muscle-specific protein of 58,487 Da. The predicted molecular weight differs significantly from the relative mobility of 86-kDa protein in reducing sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE). The full-length protein expressed in Escherichia coli also exhibits an anomalously slow mobility in SDS-PAGE; this gel retardation is a property of the N-terminal 24 kDa of the protein which contain two extended motifs of alternating alanine and proline residues, resembling the N terminus of skeletal muscle myosin light chain 1 (Nabeshima, Y. I., Fujii-Kuriyama, Y., Muramatsu, M., and Ogata, K. (1984) Nature 308, 333-338). The C-terminal 40 kDa share 49.6% sequence identity and 17% conservative substitutions with chicken skeletal muscle MyBP-C (C-protein) (Einheber, S., and Fischman, D. A. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 2157-2161). The protein contains four internal repeats of approximately 100 amino acids each, two of which bear significant resemblance to the C2 set of the immunoglobulin superfamily, and the other two are related to the type III fibronectin repeat. The arrangement of these repeats, -III-C2-III-C2-, is identical to that seen in the C-terminal 40-kDa section of MyBP-C. This repeat structure is implicated in myosin binding for the MyBP family. Finally, genomic Southern blots indicate that a single gene encodes fast skeletal muscle MyBP-H.


Day-night differences in the number of pineal "synaptic" ribbons in two diurnal rodents, the chipmunk (Tamias striatus) and the ground squirrel (Spermophilus richardsonii).
Daytime numbers of pineal "synaptic" ribbons higher than reported in the pineal gland of any other mammalian species were observed in two diurnal rodents, the eastern chipmunk and Richardson's ground squirrel. The number of "synaptic" ribbons was lower during the daytime and higher at night in both of these species.


The COVID-19 impact on reading achievement growth of Grade 3-5 students in a U.S. urban school district: variation across student characteristics and instructional modalities.
The current study aimed to explore the COVID-19 impact on reading achievement growth by Grade 3-5 students in a large urban school district in the U.S. and whether the impact differed by students' demographic characteristics and instructional modality. Specifically, using administrative data from the school district, we investigated to what extent students made gains in reading during the 2020-2021 school year relative to the pre-COVID-19 typical school year in 2018-2019. We further examined whether the effects of students' instructional modality on reading growth varied by demographic characteristics. Overall, students had lower average reading achievement gains over the 9-month 2020-2021 school year than the 2018-2019 school year with a learning loss effect size of 0.54, 0.27, and 0.28 standard deviation unit for Grade 3, 4, and 5, respectively. Substantially reduced reading gains were observed from Grade 3 students, students from high-poverty backgrounds, English learners, and students with disabilities. Additionally, findings indicate that among students with similar demographic characteristics, higher-achieving students tended to choose the fully remote instruction option, while lower-achieving students appeared to opt for in-person instruction at the beginning of the 2020-2021 school year. However, students who received in-person instruction most likely demonstrated continuous growth in reading over the school year, whereas initially higher-achieving students who received remote instruction showed stagnation or decline, particularly in the spring 2021 semester. Our findings support the notion that in-person schooling during the pandemic may serve as an equalizer for lower-achieving students, particularly from historically marginalized or vulnerable student populations.


Involvement of innate immunity in the pathogenesis of intestinal Behçet's disease.
The involvement of excessive T helper 1 (Th1) cell functions in the pathogenesis of Behçet's disease (BD) has been reported. We therefore studied Toll-like receptor (TLR)-expressing cells, which play important roles in innate immunity in patients with BD. Peripheral blood mononuclear cells (PBMC) of BD and healthy controls, and tissue specimens of intestinal BD and Crohn's disease (CD) were analysed for messenger RNA (mRNA) and protein expressions by reverse transcription-polymerase chain reaction and immunostaining respectively. PBMC of BD expressed TLR-2 and TLR-4 mRNA almost comparable with healthy controls. Intestinal lesions of BD expressed TLR-2 and TLR-4 mRNA consistently. In contrast, TLR-4 mRNA was expressed preferentially and TLR-2 mRNA was expressed less frequently in CD lesions. In intestinal samples of BD, TLR-2 and TLR-4 mRNA were detected in ileocaecal ulcer lesions, but not in unaffected sites of the same sample, indicating the association of the TLR expression with the disease manifestation of intestinal BD. TLR-2-expressing cells which were simultaneously cluster of distribution (CD)68-positive produced interleukin (IL)-12 in the lesions, indicating the participation of TLR-2-expressing cells in the Th1 skewed responses in vivo. As a possible ligand of TLR-2, in BD self-heat shock protein 60 was expressed in peripheral blood lymphocytes and intestinal tissues. Collectively, TLR-2-expressing cells as well as TLR-4-expressing cells accumulated in the intestinal lesions of BD. IL-12 produced by TLR-2-expressing cells may contribute to the induction of Th1-dominant immune responses in intestinal BD.


Toward an empirical definition of pediatric PTSD: the phenomenology of PTSD symptoms in youth.
To examine the frequency and intensity of posttraumatic stress disorder (PTSD) symptoms and their relation to clinical impairment, to examine the requirement of meeting all DSM-IV symptom cluster criteria (i.e., criteria B, C, D), and to examine the aggregation of PTSD symptom clusters across developmental stages. Fifty-nine children between the ages of 7 and 14 years with a history of trauma and PTSD symptoms were assessed with the Clinician-Administered PTSD Scale for Children and Adolescents. Data support the utility of distinguishing between the frequency and the intensity of symptoms in the investigation of the phenomenology of pediatric PTSD. Children fulfilling requirements for two symptom clusters did not differ significantly from children meeting all three cluster criteria with regard to impairment and distress. Reexperience (cluster B) showed increased aggregation with avoidance and numbing (cluster C) and hyperarousal (cluster D) in the later stages of puberty. Frequency and intensity of symptoms may both contribute to the phenomenology of pediatric PTSD. Children with subthreshold criteria for PTSD demonstrate substantial functional impairment and distress.


Measuring kinetic dissociation/association constants between Lactococcus lactis bacteria and mucins using living cell probes.
In this work we focused on quantifying adhesion between Lactococcus lactis, the model for lactic acid bacteria (LAB) and mucins. Interactions between two strains of L. lactis (IBB477 and MG1820 as control) and pig gastric mucin-based coating were measured and compared with the use of atomic force microscopy. Analysis of retraction force-distance curves shed light on the differential contributions of nonspecific and specific forces. An increased proportion of specific adhesive events was obtained for IBB477 (20% vs. 5% for the control). Blocking assays with free pig gastric mucin and its O-glycan moiety showed that oligosaccharides play a major (but not exclusive) role in L. lactis-mucins interactions. Specific interactions were analyzed in terms of kinetic constants. An increase in the loading rate of atomic force microscope tip led to a higher force between interacting biological entities, which was directly linked to the kinetic dissociation constant (K(off)). Enhancing the contact time between the tip and the sample allowed an increase in the interaction probability, which can be related to the kinetic association constant (K(on)). Variations in the loading rate and contact time enabled us to determine K(on) (3.3 × 10(2) M(-1)·s(-1)) and K(off) (0.46 s(-1)), and the latter was consistent with values given in the literature for sugar-protein interactions.


Identification of amino acid residues responsible for high initial luminescence intensity in a calcium-binding photoprotein, clytin-II.
Clytin-II (CL-II) is an isotype of the calcium-binding photoprotein clytin-I (CL-I) from Clytia gregaria. CL-II shows approximately 4.5-fold higher initial luminescence intensity than CL-I with the same luminescence capacity, and is a potential candidate for a G-protein-coupled receptor assay among photoproteins. To investigate the high initial luminescence intensity of CL-II, the chimeric proteins between CL-I and CL-II were prepared and the responsible amino acid residues in CL-II were identified by site-specific mutagenesis of CL-I. The luminescence properties of CL-I were converted to those of CL-II by the replacement of only four amino acids in CL-I, and these amino acids did not interact with 2-peroxycolenterazine.


Occult hepatitis B virus infection.
The detection of HBV DNA without HBsAg with or without the presence of HBV antibodies outside the acute phase window period defines occult HBV infection. This condition has been described in hepatocellular carcinoma (HCC), chronic hepatitis B, healthy HBV carriage and recovered infection, chronic hepatitis C and individuals without serological markers of HBV. The frequency of the diagnosis depends on the relative sensitivity of both HBsAg and HBV DNA assays. It also depends on the prevalence of HBV infection in the population. Occult HBV in blood donors has a wide range of potential origins within the natural history of the infection. It may originate from recovered infections with anti-HBs and persistent, low-level, viral replication, escape mutants undetected by the HBsAg assays or healthy chronic carriage. The last situation is mostly found with anti-HBc only. Over time, antibody markers may become undetectable leaving HBV DNA as the only marker of the infection. In all cases, the viral load is low, mostly below 10(4) IU/ml, often below 100 IU/ml. At these levels, nucleic acid testing (NAT) in pools is likely to be largely ineffective. Is occult HBV transmissible by transfusion? Carriers of anti-HBs or anti-HBc only were shown infectious in immunosuppressed organ or bone marrow transplant recipients. In immunocompetent recipients, there is no evidence that anti-HBs-containing components are infectious, even in low titre. Donations carrying anti-HBc only and HBV DNA can be infectious and this is a threat where anti-HBc is not screened. Anti-HBc screening identifies most occult HBV infection but not all. HBV NAT needs either extreme sensitivity or to be performed on individual donations to eliminate HBV DNA-containing units.


[Evaluation of the surgery results of Müller's muscle-conjunctival resection in the treatment of blepharoptosis].
To evaluate the results of Müller's muscle-conjunctival resection for correction of blepharoptosis and to discuss the advantages of this procedure. 38 patients (39 eyelids) were submitted to Müller's muscle-conjunctival resection. Blepharoptosis varied from 1.0 mm to 3.0 mm (mean: 2.0 mm). The amount of eyelid elevation produced by phenylephrine guided the amount of tissue to be resected. 33 eyelids (85%) treated with this procedure were cosmetically acceptable. Müller's muscle-conjunctival resection procedure is a relatively simple technique for blepharoptosis, with good levator function and positive 10% phenylephrine test. The advantages are: preservation of tarsus and predictable results.


Reciprocal regulation of LTA(4) hydrolase expression in human monocytes by gamma-interferon and interleukins 4 and 13: potential relevance to leukotriene regulation in glomerular disease.
Leukotriene A(4) (LTA(4)) hydrolase catalyzes the final step in the synthesis of leukotriene B(4) (LTB(4)). TH-1- and TH-2-derived cytokines may regulate LTB(4) synthesis by monocytes through their actions on the expression of LTA(4) hydrolase. Freshly isolated monocytes were incubated with pro- and anti-inflammatory cytokines for 36 h. mRNA expression was determined by Northern blot, protein expression was determined by Western blot and LTB(4) synthesis was determined by ELISA. Interferon-gamma (a TH-2-derived cytokine) increased significantly LTA(4) hydrolase mRNA expression, whereas interleukin (IL)-4 and IL-13 (both TH-2-derived cytokines) decreased LTA(4) hydrolase mRNA expression in these cells. The same effects were seen on the expression of immunoreactive LTA(4) hydrolase after incubating the monocytes with either TH-1- or TH-2-derived cytokines. The monocyte-derived cytokine IL-1 beta did not show any significant effect on LTA(4) hydrolase mRNA expression. When LTB(4) release was measured, both IL-1 beta and interferon-gamma significantly increased LTB(4) production by monocytes, while TH-2 cytokines (IL-4 and IL-13) decreased it. The opposing effects of TH-1- and TH-2-derived cytokines on the expression of LTA(4) hydrolase mRNA may regulate LTB(4) synthesis by monocytes during inflammation.


Monitoring Hydrogenation Reactions using Benchtop 2D NMR with Extraordinary Sensitivity and Spectral Resolution.
Low-field benchtop nuclear magnetic resonance (BT-NMR) spectrometers with Halbach magnets are being increasingly used in science and industry as cost-efficient tools for the monitoring of chemical reactions, including hydrogenation. However, their use of low-field magnets limits both resolution and sensitivity. In this paper, we show that it is possible to alleviate these two problems through the combination of parahydrogen-induced polarization (PHIP) and fast correlation spectroscopy with time-resolved non-uniform sampling (TR-NUS). PHIP can enhance NMR signals so that substrates are easily detectable on BT-NMR spectrometers. The interleaved acquisition of one- and two-dimensional spectra with TR-NUS provides unique insight into the consecutive moments of hydrogenation reactions, with a spectral resolution unachievable in a standard approach. We illustrate the potential of the technique with two examples: the hydrogenation of ethylphenyl propiolate and the hydrogenation of a mixture of two substrates - ethylphenyl propiolate and ethyl 2-butynoate.


Photo-CIDNP solid-state NMR on photosystems I and II:what makes P680 special?
The origin of the extraordinary high redox potential of P680, the primary electron donor of Photosystem II, is still unknown. Photochemically induced dynamic nuclear polarisation (photo-CIDNP) 13C magic-angle spinning (MAS) NMR is a powerful method to study primary electron donors. In order to reveal the electronic structure of P680, we compare new photo-CIDNP MAS NMR data of Photosystem II to those of Photosystem I. The comparison reveals that the electronic structure of the P680 radical cation is a Chl a cofactor with strong matrix interaction, while the radical cation of P700, the primary electron donor of Photosystem I, appears to be a Chl a cofactor which is essentially undisturbed. Possible forms of cofactor-matrix interactions are discussed.