Sensory rhodopsin-I as a bidirectional switch: opposite conformational changes from the same photoisomerization.
The phototaxis receptor sensory rhodopsin I (SRI) exists in two protein conformations, each of which is converted to the other by light absorption by the protein's retinylidene chromophore. One conformer inhibits a histidine-kinase attached to its bound transducer HtrI and its formation induces attractant motility responses, whereas the other conformer activates the kinase and its formation induces repellent responses. We performed Fourier transform infrared spectroscopy with temperature, pH, and mutation-induced shifts in the conformer equilibrium, and found that both conformers when present in the unphotolyzed dark state contain an all-trans retinal configuration that is photoisomerized to 13-cis, i.e., the same photoisomerization causes the opposite conformational change in the photointerconvertible pair of conformers depending on which conformer is present in the dark. Therefore, switching between the protein global conformations that define the two conformers is independent of the direction of isomerization. Insights into this phenomenon are gained from analysis of the evolution of the receptor from light-driven proton pumps, which use similar conformers for transport. The versatility of the conformational changes of microbial rhodopsins, including conformer interexchangeability in the photocycle as shown here, is likely a significant factor in the evolution of the diverse functionality of this protein family.


Parental bonding in severely suicidal adolescent inpatients.
Family environment has a clear role in suicidal behavior of adolescents. We assessed the relationship between parental bonding and suicidal behavior in suicidal (n=53) and non-suicidal (n=47) adolescent inpatients. Two dimensions of parental bonding: care and overprotection, were assessed with the Parental Bonding Instrument. Results showed that adolescents with severe suicidal behavior tended to perceive their mothers as less caring and more overprotective compared to those with mild or no suicidal behavior. A discriminant analysis distinguished significantly between adolescents with high suicidality and those with low suicidality [χ2 (5) = 15.54; p=0.01] in 71% of the cases. The perception of the quality of maternal bonding may be an important correlate of suicidal behavior in adolescence and may guide therapeutic strategies and prevention.


Miniaturized implantable cardiac monitor with a long sensing vector (BIOMONITOR III): Insertion procedure assessment, sensing performance, and home monitoring transmission success.
Implantable Cardiac Monitors (ICMs) are used for long-term monitoring of arrhythmias. BIOMONITOR III is a novel ICM with a miniaturized profile, long sensing vector due to a flexible antenna, simplified implantation with a dedicated insertion tool for pocket formation and ICM placement in a single step, and daily automatic Home Monitoring (HM) function. In 47 patients undergoing BIOMONITOR III insertion for any ICM indication, 16 investigators at 10 Australian sites assessed handling characteristics of the insertion tool, R-wave amplitudes, noise burden, P-wave visibility, and HM transmission success. Patients were followed for 1 month. All 47 attempted insertions were successful. Median time from skin incision to removal of the insertion tool after ICM insertion was 39 s (IQR 19-65) and to wound closure and cleaning was 4.7 min (IQR 3.5-7.8). All aspects of the insertion tool were rated as "good" or "excellent" in ≥97.9% and "fair" in ≤2.1% of patients, except for "force needed for tunnelling" (91.5% good/excellent, 8.5% fair). Based on HM data, R-waves in the first month were stable at 0.70 ± 0.37 mV. Median noise burden (disabling automatic rhythm evaluation) was 0.19% (IQR 0.00-0.93), equivalent to 2.7 min (IQR 0.0-13.4) per day. In HM-transmitted ECG strips with regular sinus rhythm, P-waves were visible in 89 ± 24% of heart cycles. Patient-individual automatic Home Monitoring transmission success was 98.0% ± 5.5%. The novel ICM performed well in all aspects studied, including fast insertion, reliable R-wave sensing, good P-wave visibility, and highly successful HM transmissions.


Expansion of the CCDC22 associated Ritscher-Schinzel/3C syndrome and review of the literature: Should the minimal diagnostic criteria be revised?
The Ritscher-Schinzel syndrome (RTSCS) is a rare condition with craniofacial, cardiac and fossa posterior abnormalities. RTSCS is subdivided into Ritscher-Schinzel syndrome 1 (RTSCS1) caused by pathogenic variants in coiled-coil domain-containing protein 22 (CCDC22), and Ritscher-Schinzel syndrome 2 (RTSCS2) caused by pathogenic variants in WASH complex subunit 5 (WASHC5). CCDC22 is inherited in an X-linked recessive manner while WASHC5 is inherited in an autosomal recessive manner. Only 17 individuals with a molecular diagnosis are reported. In the past, the diagnosis of RTSCS was solely based on the clinical findings, and minimal diagnostic criteria has been proposed for the syndrome: Cardiac malformations (other than isolated patent ductus arteriosis), fossa posterior malformations, and certain dysmorphic features. However, those criteria are not present in all patients. We aim to further delineate the spectrum of CDCC22 associated RTSCS and present a novel patient with epileptic encephalopathy due to a presumed disease causing CCDC22 missense variant inherited from a healthy mother and grandmother. An affected maternal uncle had passed away at the age of 12 months and was thus unavailable for genetic testing. The proband and the maternal uncle had the typical facial dysmorphism associated with RTSCS, and they closely resembled previously published RTSCS2 patients with a molecular diagnosis. This suggests that RTSCS1 and RTSCS2 patients have a similar facial gestalt. We also review the literature on RTSCS, we explore potential differences and similarities between CCDC22 and W ASHC5 associated RTSCS and discuss the minimal diagnostic criteria.


The anticancer effect of the TLR4 inhibition using TAK-242 (resatorvid) either as a single agent or in combination with chemotherapy: A novel therapeutic potential for breast cancer.
Increasing pieces of evidence indicate that inflammatory processes facilitate tumorigenesis; tumor cells simulate the mechanisms by which innate immune cells produce pro-inflammatory cytokines to exploit them for their own survival and proliferation. Toll-like receptor 4 (TLR4), which serves as one of the most well-known receptors on the surface of the immune cells, is often expressed ectopically in the tumor cells resulting in tumor progression, invasion, and chemoresistance. In this study, we examined the anticancer effects of TAK-242, a small molecule inhibitor of TLR4, on different breast cancer cell lines: MCF7, SKBR3, MDA-MB-231, and BT-474. Our results showed that the TLR4 inhibition, as revealed by the downregulation of TLR4 downstream genes, exerted desirable cytotoxicity on the TLR4-expressing cells, at least partly, through the downregulation of EGFR and c-Myc genes. TAK-242 also inhibited the proliferation of anoikis-resistant cells and suppressed the clonal growth of the indicated cells. The results of this study propose a mechanistic pathway by which the inhibition of TLR4 using TAK-242 could augment apoptotic cell death through the alteration of both nuclear factor-кB- and p53-related apoptosis genes in breast cancer cells, especially cells with overexpression of TLR4. Taken together, this study supports the idea that the activation of inflammatory pathways may have a crucial role in breast cancer progression and the inhibition of TLR4 using TAK-242, either as a single agent or in combination, seems to be a novel promising strategy that could be clinically available in foreseeable future.


Cognitive impairment in an adult Mexican population non-occupationally exposed to manganese.
We examined the association between non-occupational exposure to Mn and cognitive functions. The study was carried out in a mining district located in Hidalgo State, Mexico, with 288 adults. Air and blood Mn concentrations were determined, and neuropsychological tests were administered to explore cognitive functions and depression. Blood Mn mean was 9.5 ± 4.14 μg/L. A total of 73% of the study group were in contact with air Mn levels that surpassed the EPA recommended guideline level for non-occupational environments (0.05 μg/m(3)). Air Mn concentration was associated as a risk factor for attention impairment (OR=1.75, 95% CI: 1.01-3.06). Blood Mn levels were not associated to any of the measured outcomes. The main finding of this study is the presence of attention impairments associated to high levels of air Mn exposure. These results confirm previous studies, in which cognitive impairment is reported for exposed population.


Long-term can-sealing protection: a stable black phosphorus nanoassembly achieved through heterogeneous hydrophobic functionalization.
Black phosphorus (BP), a promising 2D material, has sparked a research boom in various areas, while its fatal atmospheric instability seriously obstructs the progress of most practical applications. To realize the novel scalable concept of can-sealing protection, the selective deposition of a series of hydrophobically- or hydrophilically-functionalized Al2O3 nanostructured capping layers has been successfully achieved to seal the top surface of the exfoliated BP flake assemblies on Ag-patterned substrates. The hydrophobic Al2O3 columnar capping is evidenced as the most promising candidate to provide comprehensive protection against the severe rapid degradation of pristine BP even under a very high-humidity environment (RH = 85%) for a long period of time. The present work provides valuable insight into the distinct anisotropic degradation of the sealed BP flake assemblies evidently induced by the deposited hydrophobically- or hydrophilically-functionalized Al2O3 capping.


[Preventive Check-ups and Routine Vaccinations in Pre-school Children - An Analysis of Mutual Participation Behaviour, Taking into Account Living Conditions].
Preventive screenings and vaccinations are essential health protecting measures in childhood, but no quantitative data on mutual distribution patterns of acceptance are available. The aim of the study was the quantification of distribution patterns and the identification of associations between preventive check-ups and defined routine vaccinations for children starting school, taking into account the social environment. The 2015 school entrance health checks included documented preventive screenings and routine vaccinations on a city-wide basis. In addition, a voluntary proxy questionnaire was distributed to persons entitled to care for the children starting school in order to record additional characteristics of family living conditions. We used SPSS 25.0 to calculate participation rates for all screenings and defined routine vaccinations as well as specific cross-prevalences and associations. Of the 407 children starting school, both prevention measures were carried out in 76.8%; additional questionnaires were distributed to those in charge of 280 (68.8%) of these children. Full preventive check-ups with complete vaccination status at the same time were observed in 70.3% of children born in Germany and 69.7% of children born in Germany with German nationality. In terms of family environment, children with siblings (17.5%), children with separated parents (13.3%) and families with up to four close relatives (7.1%) were significantly less likely than others to have completed all preventive measures. Incomplete preventive medical examinations were strongly associated with a child not born in Germany (OR=7.95) and with both parents with a non-German Nationality (OR=4.52). The distribution patterns and associations identified provide new insight for the establishment of measures specific to risk groups to increase acceptance of screenings and defined routine vaccinations. This can allow for needs-optimised health care for children based on the principle of social compensation.


The role of delta-1-pyrroline-5-carboxylate dehydrogenase (P5CDh) in the Pacific white shrimp (Litopenaeus vannamei) during biotic and abiotic stress.
Proline (Pro) metabolism is intimately associated with stress adaptation. The catabolism of Pro includes two dehydrogenation reactions catalyzed by proline dehydrogenase (ProDH) and Δ1-pyrroline-5-carboxylate dehydrogenase (P5CDh). P5CDh is a mitochondrial matrix NAD+-dependent dehydrogenase that is critical in preventing P5C-Pro intensive cycling and avoiding ROS production from electron run-off. Little is known about the roles of P5CDh in invertebrates, however. We cloned the P5CDh sequence in the Pacific white shrimp, Litopenaeus vannamei, and found that LvP5CDh is expressed predominantly in pleopod, hepatopancreas and gill. Subcellular localization analysis revealed that LvP5CDh protein was mainly found in the cytoplasm. In addition, overexpressing LvP5CDh in cells reduced ROS formation and inhibited apoptosis induced by LC50 Cd2+. Shrimp were exposed to various stress factors including infection with Vibrio alginolyticus, (½ LC50 and LC50) Cd2+, acid (pH 5.6) and alkali stress (pH 9.3). Both biotic and abiotic stress resulted in increased LvP5CDh expression and Pro accumulation; V. alginolyticus infection, pH 9.3 and LC50 Cd2+ stress apparently stimulated the Glu pathway of Pro synthesis, while pH 5.6 and ½ LC50 Cd2+ stress promoted the Orn pathway of Pro synthesis. Silencing of Lvp53 in shrimp attenuated LvP5CDh expression during Cd2+ stress, but had no effect on LvP5CDh mRNA levels if no Cd2+ stress was imposed. Our study contributes to the functional characterization of LvP5CDh in biotic and abiotic stress and reveals it to protect against ROS generation, damage to the cell, including the mitochondria, and apoptosis. Thus, LvP5CDh plays a critical role in immune defense and antioxidant responses.


Reduced erythropoietin response to anaemia in elderly patients with normocytic anaemia.
We studied the effect of age on the relationship between haemoglobin and serum erythropoietin (EPO) levels in anaemic patients. 568 patients over 70 years of age were compared with 137 patients under 70 and a reference group of 144 patients of all ages with proven iron deficiency. EPO was measured using a radioimmunoassay. We found that elderly patients with a normocytic anaemia (N = 375) had a statistically lower EPO response than younger patients with normocytic anaemia (N = 61) (p < 0.05) or patients of all ages with iron-deficiency anaemia (p < 0.05). There was no difference between the sexes. Elderly patients with microcytic or macrocytic anaemia had a normal EPO response as compared to the "gold standard" of iron deficiency. These findings suggest that a proportion of elderly patients with normocytic anaemia has an impaired EPO response.


Inhibitory Effect of Sesamolin on Melanogenesis in B16F10 Cells Determined by In Vitro and Molecular Docking Analyses.
Melanin protects the skin against the harmful effects of ultraviolet irradiation. However, melanin overproduction can result in several aesthetic problems, including melasma, freckles, age spots and chloasma. Therefore, development of anti-melanogenic agents is important for the prevention of serious hyperpigmentation diseases. Sesamolin is a lignan compound isolated from sesame seeds with several beneficial properties, including potential for melanin inhibition. The aim of this study was to evaluate the anti-melanogenic effect of sesamolin in cell culture in vitro and the underlying mechanism of inhibition using molecular docking simulation. Melanogenesis was induced by 3-isobutyl-1-methylxanthine in B16F10 melanoma cells, and the inhibitory effects of sesamolin were evaluated using zymography, a tyrosinase inhibitory activity assay, western blotting, and real-time reverse transcription-polymerase chain reaction analysis. Docking simulations between sesamolin and tyrosinase were performed using Autodock vina. Sesamolin significantly inhibited the expression of melanogenesis-related factors tyrosinase, and tyrosinase-related proteins 1 and 2 at the mRNA and protein levels. Treatment of melanoma cells with 50 µM sesamolin demonstrated the strongest inhibition against intercellular tyrosinase and melanin synthesis without exerting cytotoxic effects. Sesamolin significantly reduced mushroom tyrosinase activity in a dose-dependent manner via a competitive inhibition mechanism. Tyrosinase docking simulations supported that sesamolin (-6.5 kcal/mol) bound to the active site of tyrosinase more strongly than the positive control (arbutin, -5.7 kcal/mol). Sesamolin could be developed as a melanogenesis inhibiting agent owing to its dual function in blocking the generation of melanogenesis-related enzymes and inhibiting the enzymatic response of tyrosinase.


[Role of Chlamydia trachomatis in diseases of the urogenital system].
The authors carried out 402 examinations to detect Chlamydia trachomatis. The agent was detected by serological methods. The cultivation and serological methods proved to give significant results. Doxycyclin and Eryc capsules were successful in the therapy. The great number of positive chlamydia cases suggest to focus attention on detecting the agent on the therapy and on the prevention of the spreading of infection.


Oral disease and risk of oesophageal and gastric cancer in a nationwide nested case-control study in Sweden.
The association between the exposure to oral disease and the outcomes of oesophageal and gastric cancer was examined in a Swedish nationwide inpatient register-based nested case-control study in 1964-2008. The study included 6,156 oesophageal squamous-cell carcinoma cases that were compared with 29,993 controls, 2684 oesophageal adenocarcinoma cases that were compared with 15,036 controls and 38,308 gastric cancer cases that were compared with 99,991 controls. For oesophageal squamous cell carcinoma, the age and sex adjusted odds ratio (OR) among patients with a history of oral disease was 1.3 (95% confidence interval (95% CI): 0.9,-1.9), and 1.1 (95% CI 0.8,-1.7) after adjustment for diseases related to alcohol consumption or tobacco smoking. For oesophageal adenocarcinoma, the age and sex adjusted OR was increased (OR 1.7, 95% CI 1.1-2.6), and remained increased (OR 1.6, 95% CI 1.0-2.4) after adjustment for diseases related to smoking or alcohol consumption, gastroesophageal reflux, obesity and ulcer disease. For gastric cancer, no statistically significantly increased risk was observed (age and sex adjusted OR 0.9, 95% CI 0.7-1.1, and fully adjusted OR 0.9, 95% CI 0.7-1.1). In conclusion, this study supports the hypothesis that oral disease increases the risk of oesophageal adenocarcinoma, but not for oesophageal squamous cell carcinoma or gastric cancer. Further investigations are warranted.


Three-Dimensional Model of Hypertrophic Scar Using a Tissue-Engineering Approach.
Following wound healing, skin is replaced by a specialized tissue called scar. Sometime, this scar can become pathologic, called hypertrophic scar, with a high amount of extracellular matrix, capillaries, and myofibroblast persistence. To understand the mechanisms at the origin of the fibrosis is paramount to treat patients, but despite few animal models and in vitro studies using mainly human pathological cells cultured on plastic on monolayer, the treatment of these fibrotic scars remains unsatisfactory. As in tissue, cells are most often imbedded in extracellular matrix, we have developed, using a tissue engineering method, new in vitro models to study human fibrotic skin pathologies as hypertrophic scars. Human cells isolated from hypertrophic scars are used to reconstitute a three-dimensional fibrotic skin comprising both dermal and epidermal parts. This method called the self-assembly approach is based on the cell capacity to reconstitute their own environment as in vivo. In this chapter, the described methods include extraction and culture of human scar keratinocytes and fibroblasts from cutaneous biopsies as well as the protocols to produce fibrotic skin that can be used to study pathological process.


Dose-volume Histogram-based Predictors for Hematuria and Rectal Hemorrhage in Patients Receiving Radiotherapy After Radical Prostatectomy.
The aim of this study was to evaluate the dose-volume histogram parameters for late hematuria and rectal hemorrhage in patients receiving radiotherapy after radical prostatectomy. Data of 86 patients treated between January 2006 and June 2019 were retrospectively evaluated. The median radiation dose was 64 Gy in 32 fractions. Receiver operating characteristic (ROC) curves were used to identify optimal cut-off values for late adverse events. Eleven patients experienced hematuria, and the 5-year cumulative rate was 18%. Four patients experienced rectal hemorrhage, and the 5-year cumulative rate was 7%. ROC curve analysis demonstrated the following significant cut-off values: bladder V50 Gy: 43% (p=0.02) and V40 Gy: 50% (p=0.03) for hematuria, and rectum V60 Gy: 13% (p=0.04) and V50 Gy: 33% (p=0.03) for rectal hemorrhage. This is the first study to identify dose constraints that may reduce hematuria and rectal hemorrhage in patients receiving radiotherapy in the postoperative setting.


Effects of Gabapentin and Pregabalin on Calcium Homeostasis: Implications for Physical Rehabilitation of Musculoskeletal Tissues.
In this review, we discuss the mechanism of action of gabapentinoids and the potential consequences of long-term treatment with these drugs on the musculoskeletal system. Gabapentinoids, such as gabapentin (GBP) and pregabalin (PGB) were designed as antiepileptic reagents and are now commonly used as first-line treatment for neuropathic pain and increasingly prescribed off-label for other pain disorders such as migraines and back pain. GBP and PGB exert their analgesic actions by selectively binding the α2δ1 auxiliary subunit of voltage-sensitive calcium channels, thereby inhibiting channel function. Numerous tissues express the α2δ1 subunit where GBP and PGB can alter calcium-mediated signaling events. In tissues such as bone, muscle, and cartilage, α2δ1 has important roles in skeletal formation, mechanosensation, and normal tissue function/repair that may be affected by chronic use of gabapentinoids. Long-term use of gabapentinoids is associated with detrimental musculoskeletal outcomes, including increased fracture risk. Therefore, understanding potential complications is essential for clinicians to guide appropriate treatments.


Successful Elosulfase Alfa Desensitization Protocol in a Patient With Morquio A Syndrome.
Patients with lysosomal storage diseases may require modifications to standard drug desensitization protocols; personalized medicine as well as development of new treatment options are needed.


Biofunctionalization of polymers and their applications.
Polyhydroxyalkanoates (PHAs) are a family of biopolyesters synthesized by many types of bacteria as carbon and energy reserve materials. PHAs combine properties of thermal processibility, biodegradability, biocompatibility and sustainability. They have attracted attention from fermentation, materials and biomedical industries. Recent environmental concerns such as CO(2) emissions and plastic pollution as well as rapid exhaustion of petroleum resources have increased public and industrial interests in these unique materials. In fact, PHA has slowly evolved into an industrial value chain ranging from microbial fermentation, bioplastic packaging, biofuel, medical implants, drug delivery, protein purification, chiral chemicals and drug development. This chapter will discuss microbial PHA production and its applications in various fields.


Linking the Belowground Microbial Composition, Diversity and Activity to Soilborne Disease Suppression and Growth Promotion of Tomato Amended with Biochar.
Biochar, in addition to sequestering carbon, ameliorating soil, and improving plant performance, can impact foliar and soilborne plant diseases. Nevertheless, the mechanisms associated with suppression of soilborne diseases and improved plant performances are not well understood. This study is designed to establish the relationships between biochar-induced changes in rhizosphere microbial community structure, taxonomic and functional diversity, and activity with soilborne disease suppression and enhanced plant performance in a comprehensive fashion. Biochar suppressed Fusarium crown and root-rot of tomato and simultaneously improved tomato plant growth and physiological parameters. Furthermore, biochar reduced Fusarium root colonization and survival in soil, and increased the culturable counts of several biocontrol and plant growth promoting microorganisms. Illumina sequencing analyses of 16S rRNA gene revealed substantial differences in rhizosphere bacterial taxonomical composition between biochar-amended and non-amended treatments. Moreover, biochar amendment caused a significant increase in microbial taxonomic and functional diversity, microbial activities and an overall shift in carbon-source utilization. High microbial taxonomic and functional diversity and activity in the rhizosphere has been previously associated with suppression of diseases caused by soilborne pathogens and with plant growth promotion, and may collectively explain the significant reduction of disease and improvement in plant performance observed in the presence of biochar.


Removal of bisphenol A and methylene blue through persulfate activation by calcinated α-MnO2 nanorods: effect of ultrasonic assistance and toxicity assessment.
This work investigates the efficacy of α-MnO2 nanorods for persulfate-mediated degradation of bisphenol A (BPA) and methylene blue (MB), in silent and ultrasonic-assisted systems. The conversion of α-MnO2 nanoparticle flakes to nanorods occurs upon calcination at a temperature of 400 °C for 3 h under the ramping conditions. The comparative characterization of nanomaterials pre- and post-calcination reveals better physical, chemical, and thermal properties of α-MnO2 nanorods. The impact of various operational parameters such as pH, dosage of nanorods, persulfate dose, selected contaminant concentration, ultrasound frequency and power, scavengers, and landfill leachate medium on the degradation of pollutants is also assessed. The ultrasonic assistance yields higher removal for both BPA and MB than the silent system. This may be attributed to the generation of more radicals as ultrasound activates persulfate. This can be due to acoustic cavitation, which leads to better solute dissociation and excited state. The results obtained through scavenger tests reveal that both OH• and SO4•- can contribute to degradation, but the role of SO4•- is found dominant. Significant removal of BPA and MB ((BPA)silent, 87.12%; (MB)silent, 96.54%; (BPA)ultrasonic, 88.75%; (MB)ultrasonic, 93.86%)) is observed in landfill leachate medium. The degradation pathway for pollutants is also proposed. The toxicity of pollutants and their degradation intermediates are evaluated using Ecological Structure Activity Relationships (ECOSAR) program. The results indicate reduced toxicity of BPA intermediates, while most MB degradation intermediates show higher toxicity. Therefore, it can be affirmed that removing pollutants does not ensure a completely non-toxic process. However, the study proposes a comprehensive toxicity evaluation and eliminating toxic intermediates for completely harmless wastewater treatment.


Cystic lesions associated with meningiomas.
Three cases of meningiomas associated with large cysts are reported, and 58 other cases from the literature are reviewed. The etiology of these cysts is analyzed on the basis of surgical and pathological findings. The preoperative diagnosis of large computed tomographic hypodensities associated with meningiomas and the differential features from other cystic tumors are discussed. The surgical aspect of the solid tumor and particularly the absence of a capsule separating the tumor from the cyst are usually decisive in confirming the intratumoral nature of the cyst. In questionable cases a histologic evaluation of the cyst walls may be necessary.


Inhibition of metastasis of oral squamous cell carcinoma by anti-PLGF treatment.
Neovascularization plays a critical role in cancer metastasis. However, the molecular mechanism regulating the neovascularization in oral squamous cell carcinoma (OSCC) is poorly understood. Placental growth factor (PLGF) has been known to regulate pathological angiogenesis and has been recently shown to regulate matrix metalloproteinases (MMPs) for extracellular matrix degradation during neovascularization. Here we aimed to examine whether PLGF may regulate MMPs in the metastasis of OSCC. We found that PLGF and MMP9 levels strongly correlated in OSCC in the patients, both increased in the OSCC from the patients with metastasis of the primary OSCC. Thus, we used several human OSCC cell lines to examine the relationship between PLGF and MMP9. We found that overexpression of PLGF in OSCC cells increased expression of MMP9, while inhibition of PLGF in OSCC cells decreased expression of MMP9. However, adaptation of MMP9 levels in OSCC cells did not affect the levels of PLGF. These data suggest that PLGF may regulate MMP9 in OSCC cells, but not vice versa. Moreover, inhibition of ERK1/2, but not inhibition of PI3k or JNK pathways, substantially abolished the effect of PLGF on MMP9, suggesting that PLGF may increase expression of MMP9 via ERK/MAPK signaling pathway. Thus, our data demonstrate that PLGF-induced cancer neovascularization may be partially mediated through its effect on MMP9 activation in OSCC.


Inferring putative transmission clusters with Phydelity.
Current phylogenetic clustering approaches for identifying pathogen transmission clusters are limited by their dependency on arbitrarily defined genetic distance thresholds for within-cluster divergence. Incomplete knowledge of a pathogen's underlying dynamics often reduces the choice of distance threshold to an exploratory, ad hoc exercise that is difficult to standardise across studies. Phydelity is a new tool for the identification of transmission clusters in pathogen phylogenies. It identifies groups of sequences that are more closely related than the ensemble distribution of the phylogeny under a statistically principled and phylogeny-informed framework, without the introduction of arbitrary distance thresholds. Relative to other distance threshold- and model-based methods, Phydelity outputs clusters with higher purity and lower probability of misclassification in simulated phylogenies. Applying Phydelity to empirical datasets of hepatitis B and C virus infections showed that Phydelity identified clusters with better correspondence to individuals that are more likely to be linked by transmission events relative to other widely used non-parametric phylogenetic clustering methods without the need for parameter calibration. Phydelity is generalisable to any pathogen and can be used to identify putative direct transmission events. Phydelity is freely available at https://github.com/alvinxhan/Phydelity.


Tissue engineering approaches for bone repair: concepts and evidence.
Over the last decades, the medical world has advanced dramatically in the understanding of fracture repair. The three components needed for fracture healing are osteoconduction, osteoinduction and osteogenesis. With newly designed scaffolds, ex vivo produced growth factors and isolated stem cells, most of the challenges of critical size bone defects have been resolved in vitro, and in some cases in animal models as well. However, there are still challenges needed to be overcome before these technologies can be fully converted from the bench to the bedside. These technological and biological advancements need to be converted to mass production of affordable products that can be used in every part of the world. Vascularity, full substation of scaffolds by native bone, and bio-safety are the three most critical steps to be challenged before reaching the clinical setting.


Elective pectus bar removal following Nuss procedure for pectus excavatum: a single-institution experience.
Very few data are available on complications following elective bar removal after the Nuss procedure for pectus excavatum. The objective of this study was to investigate the data from 343 consecutive patients. From 2003 to 2009, 343 patients (85% males) had their pectus bar removed. Nine patients were excluded because of bar removal within the first year after implantation. Data were recorded from hospital records regarding: operation time, formation of callus around the bar, unilateral or bilateral incision, complications, postoperative hospital stay and if a senior resident or an intern performed the operation. The median age at the time of bar removal was 19.1 years. The median time for removal after insertion of the bar was 1139 days (range 641-2575 days). The median operation time was 34 min (range 5-183 min). The operation time depended on the formation of callus around the bar (p<0.0001), numbers of bars to be removed (p<0.0002), the need for bilateral incision (p<0.0001) and the charge of the surgeon performing the operation (p<0.0008). Eight patients (2.4%) had complications after the surgery. Five patients had pneumothorax, of which three were treated with chest tubes, and two controlled with chest X-ray. Three patients had hemothorax. Two were treated with a chest tube and the third required open surgery. Most of the patients were discharged on the day of surgery (94%) or the day after surgery (4%). Only six (2%) required more than a single day of hospitalization. Bar removal following the Nuss procedure is a quick and safe operation with very few complications. Occurrence of complications is not dependent on the experience of the surgeon.


SARS-CoV-2 Incubation Period during the Omicron BA.5-Dominant Period in Japan.
The mean virus incubation period during the SARS-CoV-2 Omicron BA.5-dominant period in Japan was 2.6 (95% CI 2.5-2.8) days, which was less than during the Delta-dominant period. Incubation period correlated with shared meals and adult infectors. A shorter incubation suggests a shorter quarantine period for BA.5 than for other variants.


Nalbuphine versus meperidine for post-operative analgesia: a double-blind comparison using the patient controlled analgesic technique.
A double-blind study was undertaken to compare nalbuphine, a synthetic partial agonist opiate, with meperidine in providing analgesia in patients following abdominal surgery, using the patient controlled analgesic technique. Both drugs showed a wide variation in demand requirements, but they were equally effective in relieving pain as assessed by the linear analogue technique. Neither drug caused a reduction of respiratory rate. The patient-controlled analgesic technique gave analgesia of good quality and it aided the comparison of the two drugs. However, some of the patients found the technique difficult to manage, and it is expensive and time-consuming. Until further experience is gained with this device its use should be confined to a constant care unit.


A longitudinal analysis of relationships between neighborhood context and underserved children's physical activity in a rapidly growing city.
Neighborhood context, which may be impacted by urban growth or residential mobility, is associated with childhood physical activity. This secondary analysis examined associations of objectively measured neighborhood characteristics with young children's moderate-to-vigorous physical activity (MVPA) and sedentary/rest time (SRT) over a period of rapid infrastructure change. Underserved preschoolers (n = 426) from a 36-month obesity prevention intervention were included in a secondary analysis (2019-2020). Based on household addresses, participants were coded as movers or non-movers and linked to four neighborhood variables: 1) distance to recreation sites, 2) annual crimes, 3) annual stray dogs, and 4) Gini index of income inequality. Accelerometry captured MVPA and SRT at baseline and 36 months. Baseline-to-follow-up neighborhood variables within moved and non-moved groups were compared. Multivariable regression assessed associations between follow-up MVPA/SRT and neighborhood variables. 45.3% of participants (n = 193) moved. Distance to the closest recreation site decreased significantly for non-movers (0.75 to 0.72 mi, p < 0.001). Nearby crimes significantly decreased for both groups (movers: 90 to 80, p < 0.001; non-movers: 77 to 74, p < 0.001) as did stray dogs (movers: 36 to 15, p < 0.001; non-movers: 36 to 18, p < 0.001). Neighborhood income inequality decreased significantly for movers (0.41 to 0.38, p = 0.03). Child MVPA minutes/day significantly decreased over time from median = 84.7 [Q1 = 64.1, Q3 = 103.9] to median = 73.6 [Q1 = 56.1, Q3 = 96.0], p < 0.001). No significant associations were detected between neighborhood variables and child physical activity. In a rapidly growing county, neighborhood context generally improved over time regardless of move status. Within this context, no associations between neighborhood characteristics and MVPA/SRT were detected in children.


A semisynthetic glycosphingolipid (LIGA20) reduces 2,4, 5-trihydroxyphenylalanine neurotoxicity in primary neuronal cultures.
The semisynthetic glycosphingolipid derivative II3Neu5-AcGgOse4-2-d-erythro-1,3-dihydroxy-2-chloro-acetamid e-4-trans- octadacene (LIGA20) attenuated injury induced by the excitotoxic L-dopa metabolite 2,4,5-trihydroxyphenylalanine (TOPA) in cultures of rat cerebellar granule cells when presented simultaneously with TOPA (EC50; 9 microM LIGA20). The natural glycosphingolipid ganglioside GM1 up to 200 microM was not neuroprotective as cotreatment, although pretreatment of cells for 2 h was efficacious. This greater potency and speed of LIGA20 action extended to limiting TOPA-induced death of cultured mesencephalic dopaminergic neurons. These data suggest that LIGA20 may have therapeutic potential for the treatment of disorders associated with excitotoxic processes.


Positive adaptation of HPA axis function in women during 44 weeks of infantry-based military training.
Basic military training (BMT) is a useful model of prolonged exposure to multiple stressors. 8-12 week BMT is associated with perturbations in the hypothalamic-pituitary-adrenal (HPA) axis which could predispose recruits to injury and psychological strain. However, characterisations of HPA axis adaptations during BMT have not been comprehensive and most studies included few if any women. We studied women undertaking an arduous, 44-week BMT programme in the UK. Anxiety, depression and resilience questionnaires, average hair cortisol concentration (HCC), morning and evening saliva cortisol and morning plasma cortisol were assessed at regular intervals throughout. A 1-h dynamic cortisol response to 1 μg adrenocorticotrophic hormone-1-24 was performed during weeks 1 and 29. Fifty-three women (aged 24 ± 2.5 years) completed the study. Questionnaires demonstrated increased depression and reduced resilience during training (F 6.93 and F 7.24, respectively, both p < 0.001). HCC increased from 3 months before training to the final 3 months of training (median (IQR) 9.63 (5.38, 16.26) versus 11.56 (6.2, 22.45) pg/mg, p = 0.003). Morning saliva cortisol increased during the first 7 weeks of training (0.44 ± 0.23 versus 0.59 ± 0.24 μg/dl p < 0.001) and decreased thereafter, with no difference between the first and final weeks (0.44 ± 0.23 versus 0.38 ± 0.21 μg/dl, p = 0.2). Evening saliva cortisol did not change. Fasting cortisol decreased during training (beginning, mid and end-training concentrations: 701 ± 134, 671 ± 158 and 561 ± 177 nmol/l, respectively, p < 0.001). Afternoon basal cortisol increased during training while there was a trend towards increased peak stimulated cortisol (177 ± 92 versus 259 ± 13 nmol/l, p = 0.003, and 589 ± 164 versus 656 ± 135, p = 0.058, respectively). These results suggest a normal stress response in early training was followed quickly by habituation, despite psychological and physical stress evidenced by questionnaire scores and HCC, respectively. There was no evidence of HPA axis maladaptation. These observations are reassuring for women undertaking arduous employment.


[Primary pigmented nodular adrenocortical disease report of 5 cases].
To study the pathology, diagnostic and therapeutic method of primary pigmented nodular adrenocortical disease (PPNAD). The data of 5 cases of PPNAD were analyzed retrospectively. Among the 5 cases, 2 were male and 3 were female. The range of age was from 12 to 53 years. All the 5 cases had symptoms of Cushing syndrome. The diagnose depended on the results of endocrine exams, ultrasound, CT, MR and pathological reports. All patients received operation of unilateral adrenalectomy. The therapeutic effects were determined by post-operative results, which concluded clinical symptoms and endocrine exams. The follow-up time was from 4 months to 3 years. All patients' symptoms of Cushing syndrome were relieved in 6 months after operation. The endocrine exam was normal in one case and obvious improved in the other four cases. Up to now, one patient drop out of the follow-up, the other 4 cases had no evidence of recurrence. PPNAD is a rare type of Cushing syndrome. Diagnose depends on endocrine exams and pathological results. Operation is the effective method for the disease.


Percutaneous techniques in the management of urological complications in renal transplant patients.
Obstruction of urine flow and urine leaks are potentially life-threatening complications in renal transplant patients. The diagnosis is difficult, and definite treatment may therefore be delayed. This paper reviews the clinical course of ten renal transplant patients with urologic complications, discusses the limitations of the standard tests used to investigate such problems, and describes the role of percutaneous antegrade pyelography and nephrostomy drainage catheter placement as safe and accurate diagnostic and therapeutic procedures.


Correction: Tuberculosis preventive treatment should be considered for all household contacts of pulmonary tuberculosis patients in India.
[This corrects the article DOI: 10.1371/journal.pone.0236743.].


SAGA and SAGA-like SLIK transcriptional coactivators are structurally and biochemically equivalent.
The SAGA-like complex SLIK is a modified version of the Spt-Ada-Gcn5-Acetyltransferase (SAGA) complex. SLIK is formed through C-terminal truncation of the Spt7 SAGA subunit, causing loss of Spt8, one of the subunits that interacts with the TATA-binding protein (TBP). SLIK and SAGA are both coactivators of RNA polymerase II transcription in yeast, and both SAGA and SLIK perform chromatin modifications. The two complexes have been speculated to uniquely contribute to transcriptional regulation, but their respective contributions are not clear. To investigate, we assayed the chromatin modifying functions of SAGA and SLIK, revealing identical kinetics on minimal substrates in vitro. We also examined the binding of SAGA and SLIK to TBP and concluded that interestingly, both protein complexes have similar affinity for TBP. Additionally, despite the loss of Spt8 and C-terminus of Spt7 in SLIK, TBP prebound to SLIK is not released in the presence of TATA-box DNA, just like TBP prebound to SAGA. Furthermore, we determined a low-resolution cryo-EM structure of SLIK, revealing a modular architecture identical to SAGA. Finally, we performed a comprehensive study of DNA-binding properties of both coactivators. Purified SAGA and SLIK both associate with ssDNA and dsDNA with high affinity (KD = 10-17 nM), and the binding is sequence-independent. In conclusion, our study shows that the cleavage of Spt7 and the absence of the Spt8 subunit in SLIK neither drive any major conformational differences in its structure compared with SAGA, nor significantly affect HAT, DUB, or DNA-binding activities in vitro.


Spatiotemporal expression of Wnt5a during the development of the striated muscle complex in rats with anorectal malformations.
Fecal incontinence and constipation after procedures for anorectal malformations (ARMs) are closely related to the maldevelopment of the striated muscle complex (SMC). Previous studies have demonstrated that myogenic regulatory factors (MRFs) play a significant role in muscle development. Wnt signal pathway is extremely important for MRFs regulation. This study was designed to investigate the spatiotemporal expression pattern of Wnt5a in SMC in ARMs rat embryos. Anorectal malformation embryos were induced by ethylene thiourea on embryonic day 10 (E10). Expression levels of protein and mRNA of Wnt5a were confirmed by immunohistochemistry staining, western blot and quantitative real-time PCR (qRT-PCR) between normal rat embryos and embryos with ARMs. Immunostaining revealed that, on embryonic day 17 (E17), the Wnt5a protein was initially expressed in the SMC in normal embryos. With the growth of pregnancy, the positive staining cells gradually increased. The same time-dependent changes of Wnt5a protein were detected in ARMs embryos. Besides, immunostaining showed that Wnt5a had a significant increase in normal embryos compared with ARMs embryos. Similarly, in Western blot and qRT-PCR, the higher expression of Wnt5a protein and mRNA were remarkable in normal embryos during the SMC development, relatively. Our study demonstrated that the downregulation of Wnt5a at the time of SMC development might partly be related to the dysplasia of SMC in ARMs.


Prevalence and Predictors of Growth Impairment and Short Stature in Pediatric-Onset Inflammatory Bowel Disease.
Growth impairment is common in children with inflammatory bowel diseases (IBD). However, the magnitude of short stature at adulthood is not well characterized. We aimed to determine the prevalence and predictors of growth impairment at diagnosis and adulthood in children with IBD. Height z-scores at diagnosis of IBD and at adulthood among 291 children with Crohn's disease (CD) and 125 with ulcerative colitis (UC) were retrieved retrospectively and compared to matched controls. Growth impairment at diagnosis was defined as height z-score for age less than or equal to -1 and short stature at adulthood as less than or equal to -2. Mean height z-score at adulthood in subjects with CD or UC was significantly different from controls although mean height did differ in males only (CD 172.3 cm ± 6.7, UC 172.7 cm ± 5.3, controls: 174.2 cm ± 7.3, p = 0.003 and p = 0.047, respectively). Diagnosis prior to final stage of puberty and male gender were risk factors for being short statured at adulthood in CD (mean difference [MD] 2.5, p = 0.013 and MD 6.25, p = 0.001, respectively) and UC (MD 4.9, p = 0.011 and MD 3.3, p = 0.034, respectively). Increased proportion of pediatric-onset IBD patients has growth impairment at adulthood. Male gender and diagnosis prior to puberty were found to impose risk for reduced adult height in both diseases.


Preparation of personalized-dose salbutamol sulphate oral films with thermal ink-jet printing.
To evaluate the use of thermal ink-jetting as a method for dosing drugs onto oral films. A Hewlett-Packard printer cartridge was modified so that aqueous drug solutions replaced the ink. The performance of the printer as a function of print solution viscosity and surface tension was determined; viscosities between 1.1 and 1.5 mm(2) s(-1) were found to be optimal, while surface tension did not affect deposition. A calibration curve for salbutamol sulphate was prepared, which demonstrated drug deposition onto an acetate film varied linearly with concentration (r(2) = 0.9992). The printer was then used to deposit salbutamol sulphate onto an oral film made of potato starch. It was found that when doses were deposited in a single pass under the print head, then the measured dose was in good agreement with the theoretical dose. With multiple passes the measured dose was always significantly less than the theoretical dose. It is proposed that the losses arise from erosion of the printed layer by shearing forces during paper handling. The losses were predictable, and the variance in dose deposited was always less than the BP limits for tablet and oral syrup salbutamol sulphate preparations. TIJ printing offers a rapid method for extemporaneous preparation of personalized-dose medicines.


Revealing the role of CCoAOMT1: fine-tuning bHLH transcription factors for optimal anther development.
The tapetum, a crucial innermost layer encompassing male reproductive cells within the anther wall, plays a pivotal role in normal pollen development. The transcription factors (TFs) bHLH010/089/091 redundantly facilitate the rapid nuclear accumulation of DYSFUNCTIONAL TAPETUM 1, a gatekeeper TF in the tapetum. Nevertheless, the regulatory mechanisms governing the activity of bHLH010/089/091 remain unknown. In this study, we reveal that caffeoyl coenzyme A O-methyltransferase 1 (CCoAOMT1) is a negative regulator affecting the nuclear localization and function of bHLH010 and bHLH089, probably through their K259 site. Our findings underscore that CCoAOMT1 promotes the nuclear export and degradation of bHLH010 and bHLH089. Intriguingly, elevated CCoAOMT1 expression resulted in defective pollen development, mirroring the phenotype observed in bhlh010 bhlh089 mutants. Moreover, our investigation revealed that the K259A mutation in the bHLH089 protein disrupted its translocation from the nucleus to the cytosol and impeded its degradation induced by CCoAOMT1. Importantly, transgenic plants with the probHLH089::bHLH089K259A construct failed to rescue proper pollen development or gene expression in bhlh010 bhlh089 mutants. Collectively, these findings emphasize the need to maintain balanced TF homeostasis for male fertility. They firmly establish CCoAOMT1 as a pivotal regulator that is instrumental in achieving equilibrium between the induction of the tapetum transcriptional network and ensuring appropriate anther development.


G protein-coupled receptors in energy homeostasis.
G-protein coupled receptors (GPCRs) compromise the largest membrane protein superfamily which play vital roles in physiological and pathophysiological processes including energy homeostasis. Moreover, they also represent the up-to-date most successful drug target. The gut hormone GPCRs, such as glucagon receptor and GLP-1 receptor, have been intensively studied for their roles in metabolism and respective drugs have developed for the treatment of metabolic diseases such as type 2 diabetes (T2D). Along with the advances of biomedical research, more GPCRs have been found to play important roles in the regulation of energy homeostasis from nutrient sensing, appetite control to glucose and fatty acid metabolism with various mechanisms. The investigation of their biological functions will not only improve our understanding of how our body keeps the balance of energy intake and expenditure, but also highlight the possible drug targets for the treatment of metabolic diseases. The present review summarizes GPCRs involved in the energy control with special emphasis on their pathophysiological roles in metabolic diseases and hopefully triggers more intensive and systematic investigations in the field so that a comprehensive network control of energy homeostasis will be revealed, and better drugs will be developed in the foreseeable future.


A Diaphragmic Traction Suture Increases Pleural Cavity Volume and Surgical Field Overview During Video-Assisted Thoracoscopic Surgery.
An elevated hemidiaphragm may impair surgical field overview during video assisted thoracoscopic surgery (VATS) and may consequently jeopardize a safe surgical procedure or prolong the duration of surgery. The aim of this study was to evaluate if tension applied to a diaphragmatic suture improves the surgical field overview. Following informed consent and at the surgeon's discretion during elective VATS procedures, a single stitch was placed at the posterior tendinous border of the diaphragm and retracted through the camera port. The surgical field overview was evaluated using a numeric rating scale (1-10) by the surgeon before and after applying tension during the procedure, and later by 9 VATS surgeons (> 10 years' experience) using video recordings. During a 4-month period, 43 patients scheduled for elective VATS by two surgeons gave informed consent to participate. The hemidiaphragm was elevated to such an extent in 27 patients that the surgeon placed a diaphragmatic stitch suture. When tension was applied to the suture, surgical field overview improved significantly (p < 0.001). A diaphragmatic traction suture improves surgical field overview in selected patients with elevation of the hemidiaphragm. This simple procedure may facilitate VATS in patients with impaired surgical field overview and consequently improve safety during VATS. CLINICAL TRIAL NUMBER: http://ClinicalTrials.gov (No. NCT04837950).


Indirect adjustment for multiple missing variables applicable to environmental epidemiology.
Develop statistical methods for survival models to indirectly adjust hazard ratios of environmental exposures for missing risk factors. A partitioned regression approach for linear models is applied to time to event survival analyses of cohort study data. Information on the correlation between observed and missing risk factors is obtained from ancillary data sources such as national health surveys. The relationship between the missing risk factors and survival is obtained from previously published studies. We first evaluated the methodology using simulations, by considering the Weibull survival distribution for a proportional hazards regression model with varied baseline functions, correlations between an adjusted variable and an adjustment variable as well as selected censoring rates. Then we illustrate the method in a large, representative Canadian cohort of the association between concentrations of ambient fine particulate matter and mortality from ischemic heart disease. Indirect adjustment for cigarette smoking habits and obesity increased the fine particulate matter-ischemic heart disease association by 3%-123%, depending on the number of variables considered in the adjustment model due to the negative correlation between these two risk factors and ambient air pollution concentrations in Canada. The simulations suggested that the method yielded small relative bias (<40%) for most cohort designs encountered in environmental epidemiology. This method can accommodate adjustment for multiple missing risk factors simultaneously while accounting for the associations between observed and missing risk factors and between missing risk factors and health endpoints.


Stress through the mind of the beholder: preliminary differences in child and maternal perceptions of child stress in relation to child cortisol and cardiovascular activity.
The present study examined associations among parent and child reports of youth's stressful life events (SLEs), perceived stress, and biological measures of stress activity (i.e. cortisol and cardiovascular activity). Examining these aspects of youth stress presents several challenges. Unlike adult studies of individual differences in which information regarding SLEs, perceptions of events, and biological activity are gathered from one individual, assessment of individual differences among children usually involves other informants (e.g. parent). However, parent and child reports of SLEs and the child's psychological response to such events are often discordant. Moreover, examinations of youth perception of stress are hampered by limitations of child cognitive processes, as well as parents' limited knowledge of their child's perception of stress. In a preliminary effort to unscramble the complex effects of youth SLEs and perceived stress in relation to biological response to acute stressors, this study examined 51 boys and girls aged 7-16, with no history of psychopathology or medical concerns. Contrary to hypotheses, findings revealed that compared to actual experiences of stress, perceived stress has greater associations with both cortisol and cardiovascular activity. That is, perceived stress is more biologically salient relative to actual stress. Results also suggest that informant differences may explain some previous inconsistent findings in studies of youth's stress reactivity. The current findings mirror the adult studies that show appraisal and perception of traumatic and stressful events may be more predictive of negative health and mental health outcomes than the severity of the events. Further studies are needed to understand the impact of youth's perceptions of stress on their biological stress reactions and later health outcomes such as clinical disorders.


Fluorescent porous organic cage with good water solubility for ratiometric sensing of gold(III) ion in aqueous solution.
Fluorescent porous organic cage with good water solubility is of great interest but still challenging for its fluorescent sensing application. Poor water solubility and single signal of most previous probes are unfavorable for the monitoring of Au3+ generated from the potential dissociation of gold nanoparticles in environmental and biological samples. Here we report a water-soluble porous organic cage as a ratiometric fluorescent probe for Au3+ in aqueous solution. The prepared porous organic cage with good water solubility showed specific redox interaction with Au3+ in pH 5, leading to the change of dual emission at 420 and 484 nm. Based on the change of fluorescence ratio, a simple ratiometric sensing method for Au3+ from the dissociation of gold nanoparticles in aqueous solution was developed. The proposed method gave a calibration function of F484/F420 = 0.0370[Au3+] + 0.5689 (where F484/F420 is the intensity ratio of fluorescence at 484 nm to that at 420 nm; [Au3+] in μM) (R2 = 0.9975) in the concentration range of 1-60 μM, the limit of detection (3s) of 8 nM, and the relative standard deviation of 0.26% for 10 replicate detections of 50 μM Au3+. The recoveries of spiked Au3+ in domestic wastewater and human serum samples ranged from 94.66% to 105.61%.


Special Issue: Bacteriophage Treatment as an Alternative Technology to Inactivate Pathogenic Bacteria: A Generalized Worldwide Growing Acceptance.
The increasing worldwide rate of antibiotic resistance has led to a higher incidence of bacterial infections that require alternative methods for their control not only in human medicine, but also in other areas, such as in veterinary medicine, agro-food field and wastewater treatment [...].


Cancer-associated fibroblast-like fibroblasts in vocal fold leukoplakia suppress CD8+T cell functions by inducing IL-6 autocrine loop and interacting with Th17 cells.
The characteristics of fibroblast cells in head and neck precancerous lesion and its ability to secrete inflammatory cytokines and affect CD8+T cell functions remain unclear. Herein, we reported the existence of fibroblasts in human-derived vocal fold leukoplakia (VFL) with positive staining of fibroblast activation protein (FAP) and α-smooth muscle actin (α-SMA). The fibroblasts from VFL and cancer-associated fibroblasts (CAFs) from head and neck squamous cell carcinoma (HNSCC) displayed similar cellular functions and robust inflammatory cytokine secretions. The effects of fibroblasts from VFL in inducing the apoptosis, depletion of CD8+ T cells and recruitment of regulatory T cells (Treg cells) were observed. We further assessed the autocrine loop within VFL fibroblasts to self-stimulate by secreting IL-6, TGF-β through the IL-6/JAK2/STAT3 pathway. The synergistic stimulation of IL-6 and TGF-β promoted Th17 cell differentiation and IL-17A secretion, which could result in fibroblast activation in another positive loop. Tocilizumab (TOC), a monoclonal antibody targeting IL-6R, managed to suppress the overexpression of both IL-6 and TGF-β in VFL fibroblasts, and thus blocking IL-6 autocrine loop and CAF-Th17 loop in vitro. In a murine model of oral leukoplakia (OL), local injection of TOC inhibited the outgrowth of lesions and showed notable effect in control of OL progression in vivo. Our findings establish a novel rationale for blocking the IL-6/JAK2/STAT3 pathway to inhibit vocal fold (oral) leukoplakia progression and postpone HNSCC tumorigenesis.


Bidirectional role of microtubule dynamics in the acquisition and maintenance of temporal information in dorsolateral striatum.
Accurate and precise timing is crucial for complex and purposeful behaviors, such as foraging for food or playing a musical instrument. The brain is capable of processing temporal information in a coordinated manner, as if it contains an 'internal clock'. Similar to the need for the brain to orient itself in space in order to understand its surroundings, temporal orientation and tracking is an essential component of cognition as well. While there have been multiple models explaining the neural correlates of timing, independent lines of research appear to converge on the conclusion that populations of neurons in the dorsal striatum encode information relating to where a subject is in time relative to an anticipated goal. Similar to other learning processes, acquisition and maintenance of this temporal information is dependent on synaptic plasticity. Microtubules are cytoskeletal proteins that have been implicated in synaptic plasticity mechanisms and therefore are considered key elements in learning and memory. In this study, we investigated the role of microtubule dynamics in temporal learning by local infusions of microtubule stabilizing and destabilizing agents into the dorsolateral striatum. Our results suggested a bidirectional role for microtubules in timing, such that microtubule stabilization improves the maintenance of learned target durations, but impairs the acquisition of a novel duration. On the other hand, microtubule destabilization enhances the acquisition of novel target durations, while compromising the maintenance of previously learned durations. These findings suggest that microtubule dynamics plays an important role in synaptic plasticity mechanisms in the dorsolateral striatum, which in turn modulates temporal learning and time perception.


Pap smear outcomes in elderly women living with HIV and HIV-negative matched controls.
To describe risk factors/incidence of abnormal cervical/vaginal cytology/histology and cancer among women living with human immunodeficiency virus (WLHIV) ≥65 years compared to HIV-negative matched controls. Retrospective cohort of patients who underwent Pap screening at the University of Maryland 01/2003-04/2019. WLHIV and HIV-negative controls (n = 70 each) underwent 140/151 Pap tests, respectively. Among WLHIV, 29% exhibited abnormal results and were less likely than HIV-negative women with normal Paps to have had serially negative Pap tests prior to age 65 (p = .03). In both groups, 1.4% developed cervical cancer. Abnormal Paps were more frequent in WLHIV than in HIV-negative women (31% vs 10%, p < .0001, RR:3.2, 95%CI1.9-5.4) as was HRHPV (high-risk human papillomavirus) status (43% vs 19%, p = .0233, RR:2.3, 95%CI1.2-4.6). The RR for an abnormal Pap was 2.6 (95% CI:1.1-4.2) for VL >1000 copies/mL and 0.4 (95% CI:0.2-0.7) for CD4 count of >200 cells/μL. No individual with an initially normal Pap experienced an abnormal result over a mean of 42.5 and 43.5 months in the HIV-positive and HIV-negative groups, respectively. HIV status was associated with a higher rate of abnormal Pap/HRHPV; however, no significant difference in cervical/vaginal cancer. Elevated VL/low CD4 count were associated with greater risk for an abnormal Pap.


Racial Differences in Postpartum Blood Pressure Trajectories Among Women After a Hypertensive Disorder of Pregnancy.
Maternal morbidity and mortality are increasing in the United States, most of which occur post partum, with significant racial disparities, particularly associated with hypertensive disorders of pregnancy. Blood pressure trajectory after a hypertensive disorder of pregnancy has not been previously described. To describe the blood pressure trajectory in the first 6 weeks post partum after a hypertensive disorder of pregnancy and to evaluate whether blood pressure trajectories differ by self-reported race. This prospective cohort study included deliveries between January 1, 2018, and December 31, 2019. Women with a clinical diagnosis of a hypertensive disorder of pregnancy were enrolled in a postpartum remote blood pressure monitoring program at the time of delivery and were followed up for 6 weeks. Statistical analysis was performed from April 6 to 17, 2020. Mixed-effects regression models were used to display blood pressure trajectories in the first 6 weeks post partum. A total of 1077 women were included (mean [SD] age, 30.2 [5.6] years; 804 of 1017 White [79.1%] and 213 of 1017 Black [20.9%]). Systolic and diastolic blood pressures were found to decrease rapidly in the first 3 weeks post partum, with subsequent stabilization (at 6 days post partum: mean [SD] peak systolic blood pressure, 146 [13] mm Hg; mean [SD] peak diastolic blood pressure, 95 [10] mm Hg; and at 3 weeks post partum: mean [SD] peak systolic blood pressure, 130 [12] mm Hg; mean [SD] peak diastolic blood pressure, 85 [9] mm Hg). A significant difference was seen in blood pressure trajectory by race, with both systolic and diastolic blood pressure decreasing more slowly among Black women compared with White women (mean [SD] peak systolic blood pressure at 1 week post partum: White women, 143 [14] mm Hg vs Black women, 146 [13] mm Hg; P = .01; mean [SD] peak diastolic blood pressure at 1 week post partum: White women, 92 [9] mm Hg vs Black women, 94 [9] mm Hg; P = .02; and mean [SD] peak systolic blood pressure at 3 weeks post partum: White women, 129 [11] mm Hg vs Black women, 136 [15] mm Hg; P < .001; mean [SD] peak diastolic blood pressure at 3 weeks post partum: White women, 84 [8] mm Hg vs Black women, 91 [13] mm Hg; P < .001). At the conclusion of the program, 126 of 185 Black women (68.1%) compared with 393 of 764 White women (51.4%) met the criteria for stage 1 or stage 2 hypertension (P < .001). This study found that, in the postpartum period, blood pressure decreased rapidly in the first 3 weeks and subsequently stabilized. The study also found that, compared with White women, Black women had a less rapid decrease in blood pressure, resulting in higher blood pressure by the end of a 6-week program. Given the number of women with persistent hypertension at the conclusion of the program, these findings also appear to support the importance of ongoing postpartum care beyond the first 6 weeks after delivery.


Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy in Young Patients With Peritoneal Metastasis of Colorectal Cancer-An Asian Experience.
Cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) are accepted treatment options for patients with cancers associated with the abdominal region. Currently, there are insufficient data that evaluate the effectiveness of these therapeutic options in young patients who have peritoneal metastasis (PM) originating from colorectal cancer. In this investigation, we evaluated perioperative data, clinicopathological manifestations, and survival consequences in 46 younger (≤50 y) patients who underwent CRS and HIPEC therapy between 2017 June and 2019 June and then compared these data with 94 older patients (aged >50 y). Compared with older patients, younger patients had a higher synchronous PM incidence (78.3% versus 51.1%, P < 0.001) and were more likely to exhibit signet ring histology and mucinous (29.8% versus 60.9%; P < 0.001). The cancer-specific survival rates after CRS and HIPEC treatment were similar in both age sets. Multivariate Cox regression revealed that mucinous/signet adenocarcinoma (hazard ratio 2.20, 95% confidence interval 1.02-4.74; P = 0.044) and rectal origin (hazard ratio 2.51, 95% confidence interval 1.11-5.67; P = 0.027) were independent risk factors for a lower cancer-specific survival rate. Younger (age ≤50 y) patients who have PM of colorectal cancer origin often present synchronous PM, which is less commonly observed in older patients. Tumors in younger patients are more aggressive; however, post-CRS and HIPEC treatment, the benefits are similar to those observed in older cohorts.


Rapid killing of single neurons by irradiation of intracellularly injected dye.
A simple technique for rapidly killing all or part of single neurons consists of filling the cell with Lucifer Yellow CH and irradiating all or part of it with intense blue light. Such treatment kills the irradiated part of the cell within a few minutes. Adjacent cells are not affected.


The role of sexual orientation and the perceived threat posed by men in the acceptance of sexism.
We aimed at examining the differences between straight (N = 583) and gay (N = 112) people in terms of the acceptance of ambivalent sexist attitudes not only toward women (ASI) but also toward men (AMI) and how the level of perceived threat posed by men (Realistic Threat Scale) influences these attitudes' acceptance. The study results showed that gay men endorsed hostile sexism at the same level as straight men, while lesbian women were more hostile toward men than straight women. In this study, gay people manifested significantly lower levels of benevolent (sexist) attitudes toward both genders than straight people. While a positive relationship was identified between the perceived threat posed by men and hostility toward men, there was a reciprocal correlation in the case of hostile sexism. Men who recognized the threat men can pose to women were less accepting of hostile sexism and more accepting of hostile attitudes toward their own gender group. Moreover, straight men who recognized this threat endorsed less benevolent attitudes toward their own gender group than those who failed to admit it. Our results showed that heterosexual interdependence and recognizing the threat posed by men highly influence the extent to which hostile and benevolent (sexist) attitudes toward women and men are accepted or rejected. The implications and practical relevance of our study are also discussed.


Determination of remoxipride in human plasma and urine by reversed-phase ion-pair high-performance liquid chromatography.
A sensitive method is described for the measurement of remoxipride in human plasma and urine. Remoxipride and its internal standard are extracted from plasma or urine at pH 12 with a mixture of hexane and methyl tert.-butyl ether. After washing the organic phase with base, the compounds are extracted into acid and analyzed on a C18 column with ultraviolet detection at 214 nm. The mobile phase is composed of acetonitrile and aqueous buffer (sodium perchlorate and phosphoric acid, pH 1.7). The limits of reliable quantitation for remoxipride are 12.5 and 50 ng/ml for plasma and urine, respectively. The run times are 6 min for plasma and 3 min for urine. The method has been successfully used to assay remoxipride clinical study samples. This mobile phase has also been successfully applied to the analysis of other basic drugs such as cimetidine, codeine, diltiazem and quinidine with minor modifications.


Efficient separation and recovery of lithium through volatilization in the recycling process of spent lithium-ion batteries.
Spent lithium-ion batteries (LIBs) comprise different kinds of valuable metals with recovery and reuse value. Aiming to address the difficulty of recycling lithium from spent LIBs through conventional pyrometallurgy, a new method of high-efficiency separation and recovery of lithium through volatilization is proposed. In this new method, spent LIBs as the raw material, copper slag as the only flux and CaCl2 as an additive are utilized to recover lithium from spent LIBs. Under the optimal conditions, the volatilization rate of Li was 96.87%. During the smelting process, lithium is volatilized into the gas phase in the form of LiCl, where lithium can be recycled from the dust. In light of the experimental results, the addition of CaCl2 contributes to the formation of LiCl. The kinetics study showed that the volatilization of LiCl was controlled by an interfacial chemical reaction, and the apparent activation energy was 42.57 kJ/mol. In addition, Li2CO3 could be obtained from lithium-containing dust using a precipitation process. This method achieves efficient separation of lithium during the reduction smelting process. The phase transformation and kinetics of the separation process were investigated, and reaction mechanism was revealed. Importantly, the novel process provides new ideas and perspectives for the separation of lithium from spent LIBs through a pyrometallurgical process.


Impact of local COVID-19 alert levels on rhegmatogenous retinal detachment.
To investigate the impact of the local alert levels regarding coronavirus disease 2019 (COVID-19) on the clinical patterns of rhegmatogenous retinal detachment (RRD) in Japan. Retrospective, single-center, consecutive case series. We compared two groups of RRD patients, a COVID-19 pandemic group and a control group. Based on the local alert levels in Nagano, five periods during the COVID-19 pandemic were further analyzed: epidemic 1 (state of emergency), inter-epidemic 1, epidemic 2 (second epidemic duration), inter-epidemic 2, and epidemic 3 (third epidemic duration). Patients' characteristics, including symptoms' duration before visiting our hospital, macula status, and retinal detachment (RD) recurrence rate in each period, were compared with those in a control group. There were 78 patients in the pandemic group and 208 in the control group. The pandemic group had a longer duration of symptoms than the control group (12.0 ± 13.5 days vs. 8.9 ± 14.7 days, P = 0.0045). During the epidemic 1 period, patients had a higher rate of macula-off RRD (71.4% vs. 48.6%) and RD recurrence (28.6% vs. 4.8%) than the control group. This period also demonstrated the highest rates compared to all other periods in the pandemic group. During the COVID-19 pandemic, RRD patients significantly delayed visiting a surgical facility. They showed a higher rate of macula-off and recurrence compared to the control group during the state of emergency than during other periods of the COVID-19 pandemic, although the difference was not statistically significant due to the small sample size.


Hypoxia mimetics restore bone biomineralisation in hyperglycaemic environments.
Diabetic patients have an increased risk of fracture and an increased occurrence of impaired fracture healing. Diabetic and hyperglycaemic conditions have been shown to impair the cellular response to hypoxia, via an inhibited hypoxia inducible factor (HIF)-1α pathway. We investigated, using an in vitro hyperglycaemia bone tissue engineering model (and a multidisciplinary bone characterisation approach), the differing effects of glucose levels, hypoxia and chemicals known to stabilise HIF-1α (CoCl2 and DMOG) on bone formation. Hypoxia (1% O2) inhibited bone nodule formation and resulted in discrete biomineralisation as opposed to the mineralised extracellular collagen fibres found in normoxia (20% O2). Unlike hypoxia, the use of hypoxia mimetics did not prevent nodule formation in normal glucose level. Hyperglycaemic conditions (25 mM and 50 mM glucose) inhibited biomineralisation. Interestingly, both hypoxia mimetics (CoCl2 and DMOG) partly restored hyperglycaemia inhibited bone nodule formation. These results highlight the difference in osteoblast responses between hypoxia mimetics and actual hypoxia and suggests a role of HIF-1α stabilisation in bone biomineralisation that extends that of promoting neovascularisation, or other system effects associated with hypoxia and bone regeneration in vivo. This study demonstrates that targeting the HIF pathway may represent a promising strategy for bone regeneration in diabetic patients.


Lysophosphatidic acid is a lipid mediator with wide range of biological activities. Biosynthetic pathways and mechanism of action.
Lysophosphatidic acid (LPA) is a lipid mediator required for maintaining homeostasis of numerous physiological functions and also involved in development of some pathological processes through interactions with G protein-coupled receptors. Recently many data have appeared about the role of this phospholipid in humans, but pathways of LPA biosynthesis and mechanisms of its action remain unclear. This review presents modern concepts about biosynthesis, reception, and biological activity of LPA in humans. Natural and synthetic LPA analogs are considered in the view of their possible use in pharmacology as agonists and/or antagonists of G protein-coupled receptors of LPA.


Broken silence restored--remodeling primes for deacetylation at replication forks.
Faithful propagation of chromatin structures requires assimilation of new histones to the modification profile of individual loci. In this issue of Molecular Cell, Rowbotham and colleagues identify a remodeler, SMARCAD1, acting at replication sites to facilitate histone deacetylation and restoration of silencing.


A dual inhibitor of matrix metalloproteinases and a disintegrin and metalloproteinases, [¹⁸F]FB-ML5, as a molecular probe for non-invasive MMP/ADAM-targeted imaging.
Numerous clinical studies have shown a correlation between increased matrix metalloproteinase (MMP)/a disintegrin and metalloproteinase (ADAM) activity and poor outcome of cancer. Various MMP inhibitors (MMPIs) have been developed for therapeutic purposes in oncology. In addition, molecular imaging of MMP/ADAM levels in vivo would allow the diagnosis of tumors. We selected the dual inhibitor of MMPs and ADAMs, ML5, which is a hydroxamate-based inhibitor with affinities for many MMPs and ADAMs. ML5 was radiolabelled with (18)F and the newly obtained radiolabelled inhibitor was evaluated in vitro and in vivo. ML5 was radiolabelled by direct acylation with N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB) for PET (positron emission tomography). The resulting radiotracer [(18)F]FB-ML5 was evaluated in vitro in human bronchial epithelium 16HBE cells and breast cancer MCF-7 cells. The non-radioactive probe FB-ML5 and native ML5 were tested in a fluorogenic inhibition assay against MMP-2, -9, -12 and ADAM-17. The in vivo kinetics of [(18)F]FB-ML5 were examined in a HT1080 tumor-bearing mouse model. Specificity of probe binding was examined by co-injection of 0 or 2.5mg/kg ML5. ML5 and FB-ML5 showed high affinity for MMP-2, -9, -12 and ADAM-17; indeed IC50 values were respectively 7.4 ± 2.0, 19.5 ± 2.8, 2.0 ± 0.2 and 5.7 ± 2.2 nM and 12.5 ± 3.1, 31.5 ± 13.7, 138.0 ± 10.9 and 24.7 ± 2.8 nM. Radiochemical yield of HPLC-purified [(18)F]FB-ML5 was 13-16% (corrected for decay). Cellular binding of [(18)F]FB-ML5 was reduced by 36.6% and 27.5% in MCF-7 and 16 HBE cells, respectively, after co-incubation with 10 μM of ML5. In microPET scans, HT1080 tumors exhibited a low and homogeneous uptake of the tracer. Tumors of mice injected with [(18)F]FB-ML5 showed a SUVmean of 0.145 ± 0.064 (n=6) which decreased to 0.041 ± 0.027 (n=6) after target blocking (p<0.05). Ex vivo biodistribution showed a rapid excretion through the kidneys and the liver. Metabolite assays indicated that the parent tracer represented 23.2 ± 7.3% (n=2) of total radioactivity in plasma, at 90 min post injection (p.i.). The nanomolar affinity MMP/ADAM inhibitor ML5 was successfully labelled with (18)F. [(18)F]FB-ML5 demonstrated rather low binding in ADAM-17 overexpressing cell lines. [(18)F]FB-ML5 uptake showed significant reduction in the HT1080 tumor in vivo after co-injection of ML5. [(18)F]FB-ML5 may be suitable for the visualization/quantification of diseases overexpressing simultaneously MMPs and ADAMs.


Pharmacological control of the in vivo passive anaphylactic shock by the PAF-acether antagonist compound BN 52021.
BN 52021 antagonized PAF-acether-induced bronchoconstriction (BC) in the guinea-pig and inhibited BC triggered by antigen in passively sensitized animals. The anti-anaphylactic activity was prevented by propranolol and may either result from an additional property of the drug, independent from PAF antagonism or from different PAF-dependent mechanisms with different responsiveness to the various antagonists.


Advances in Treatment of Post-Traumatic Stress Disorder with Chinese Medicine.
"Timely, near, and expectation" is the main principle of battlefield rescue for military combat stress reaction (CSR). Post-traumatic stress disorder (PTSD) is the most common form of CSR and a long-term persistent mental disorder that is caused by unusual threatening or catastrophic psychological trauma. Chinese medicine (CM) has abundant resources, is simple, easy to master, with few side effects. This article summarizes the cellular and animal experimental mechanisms of CM treatment on PTSD, suggesting that traditional Chinese herbs and acupuncture can protect brain functional areas, and adjust hypothalamus-pituitary-adrenal axis. Traditional Chinese herbs and acupuncture have shown good anti-stress efficacy and fewer side effects in clinical application, which may improve the CSR in the battlefield.


Rationally Designed Dual Base Pair Mismatch Enables Toehold-Mediated Strand Displacement to Efficiently Recognize Single-Nucleotide Polymorphism without Enzymes.
The efficiency of the enzyme-free toehold-mediated strand displacement (TMSD) technique is often insufficient to detect single-nucleotide polymorphism (SNP) that possesses only single base pair mismatch discrimination. Here, we report a novel dual base pair mismatch strategy enabling TMSD biosensing for SNP detection under enzyme-free conditions when coupled with catalytic hairpin assembly (CHA) and fluorescence resonance energy transfer (FRET). The strategy is based on a competitive strand displacement reaction mechanism, affected by the thermodynamic stability originating from rationally designed dual base pair mismatch, for the specific recognition of mutant-type DNA. In particular, enzyme-free nucleic acid circuits, such as CHA, emerge as a powerful method for signal amplification. Eventually, the signal transduction of this proposed biosensor was determined by FRET between streptavidin-coated 605 nm emission quantum dots (605QDs, donor) and Cy5/biotin hybridization (acceptor, from CHA) when incubated with each other. The proposed biosensor displayed high sensitivity to the mutant target (MT) with a detection concentration down to 4.3 fM and led to high discrimination factors for all types of mismatches in multiple sequence contexts. As such, the application of this proposed biosensor to investigate mechanisms of the competitive strand displacement reaction further illustrates the versatility of our dual base pair mismatch strategy, which can be utilized for the creation of a new class of biosensors.


Effects of Folic Acid Combined with DHA Supplementation on Cognitive Function and Amyloid-β-Related Biomarkers in Older Adults with Mild Cognitive Impairment by a Randomized, Double Blind, Placebo-Controlled Trial.
The neuroprotective benefits of combined folic acid and docosahexaenoic acid (DHA) on cognitive function in mild cognitive impairment (MCI) patients are suggested but unconfirmed. To explore the effects of 6-month folic acid + DHA on cognitive function in patients with MCI. Our randomized controlled trial (trial number ChiCTR-IOR-16008351) was conducted in Tianjin, China. We divided 160 MCI patients aged > 60 years into four regimen groups randomly: folic acid (0.8 mg/day) + DHA (800 mg/day), folic acid (0.8 mg/day), DHA (800 mg/day), and placebo, for 6 months. Cognitive function and blood amyloid-β peptide (Aβ) biomarker levels were measured at baseline and 6 months. Cognitive function was also measured at 12 months. A total of 138 patients completed this trial. Folic acid improved the full-scale intelligence quotient (FSIQ), arithmetic, and picture complement scores; DHA improved the FSIQ, information, arithmetic, and digit span scores; folic acid + DHA improved the arithmetic (difference 1.67, 95% CI 1.02 to 2.31) and digital span (1.33, 0.24 to 2.43) scores compared to placebo. At 12 months, all scores declined in the intervention groups. Folic acid and folic acid + DHA increased blood folate (folic acid + DHA: 7.70, 3.81 to 11.59) and S-adenosylmethionine (23.93, 1.86 to 46.00) levels and reduced homocysteine levels (-6.51, -10.57 to -2.45) compared to placebo. DHA lower the Aβ40 levels (-40.57, -79.79 to -1.35) compared to placebo (p < 0.05), and folic acid + DHA reduced the Aβ42 (-95.59, -150.76 to -40.43) and Aβ40 levels (-45.75, -84.67 to -6.84) more than DHA (p < 0.05). Folic acid and DHA improve cognitive function and reduce blood Aβ production in MCI patients. Combination therapy may be more beneficial in reducing blood Aβ-related biomarkers.


Background and design of a controlled clinical trial (ARC 090) for the treatment of opioid dependence.
This study represents the largest clinical trial reported to date that demonstrated the efficacy of buprenorphine for opioid dependence treatment (Johnson et al. 1992). Although the study design was adequate to demonstrate differences between treatment groups, there has not been a consensus regarding the most appropriate method for analyzing various outcome measures of this and similar studies. To present a comprehensive review of these methods, other chapters in this monograph focus on various analytical techniques for assessing one of these measures--urine toxicology screens--for illicit opioids.


Severe right hip pain.
While this patient had osteoporosis, she hadn't experienced any recent trauma or falls that would explain her pain.


Rhizoma drynariae total flavonoids inhibit the inflammatory response and matrix degeneration via MAPK pathway in a rat degenerative cervical intervertebral disc model.
Rhizoma drynariae total flavonoids (RDTF) are extracted from Drynaria fortunei J. Sm (D. fortunei), which was a Chinese herb commonly used to treat fractures and bruises. Modern pharmacological studies indicate flavonoids have anti-inflammatory effect in clinical practice. However, its active ingredients and the mechanisms of action are far from clear. The present study aims to determine whether RDTF can protect against intervertebral disc degeneration in a rat cervical intervertebral disc model and investigate the associated molecular mechanisms. Sprague Dawley (SD) rats were randomized into five groups: control group (CG, n = 8), intervertebral disc degeneration group (NG, n = 8), low-dose RDTF-treated group (LG, n = 8), medium-dose RDTF-treated group (MG, n = 8), and high-dose RDTF-treated group (HG, n = 8). Hematoxylin and eosin (HE) staining, immunohistochemistry (IHC), immunofluorescence, ELISA, Western blot and quantitative real time PCR (qRT-PCR) assays were used to investigate inflammatory, catabolic factors and the latent regulatory mechanism of the effects of RDTF on intervertebral disc cells. HE staining showed disc degeneration in all groups except CG, and the function was restored after RDTF treatment. IHC, Western blot, qRT-PCR, immunofluorescence and ELISA results showed that RDTF prevented intervertebral disc degeneration by suppressing mitogen-activated protein kinase (MAPK) pathway, which reduced expression of intracellular matrix metalloproteinases (MMPs), such MMP3, MMP13, and inflammatory factors including interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α). Notably RDTF inhibited extracellular matrix (ECM) degeneration by increasing expression of aggrecan and collagen type II and preventing the upregulation of collagen type I and III. It suggests that RDTF has a potential therapeutic effect on cervical spondylosis.


Effect of central sensitization inventory on the number of painful sites and pain severity in a Japanese regional population cohort.
To investigate the association between the central sensitization inventory (CSI), a screening tool for central sensitization, and the number of painful sites and the severity of pain in locomotive organs in an epidemiological study in the elderly. A total of 379 individuals who underwent musculoskeletal disease screening were enrolled in this study. The CSI was used to assess symptoms of central sensitization. The number and location of painful sites and the severity of pain were evaluated using pain mapping and a numerical rating scale (NRS) at 37 sites. We investigated the association between the number of painful sites and CSI score, and the association between the severity of low back pain or knee pain and CSI score. There was a positive correlation between CSI score and the number of painful sites. The CSI score was significantly higher in those with significant low back pain than in those without pain, and the high-CSI group tended to have a greater number of painful sites. Comparison of CSI scores between participants with low back pain alone and those with low back pain and posterior lower leg pain showed that the latter group had a significantly higher CSI score than the former group. The CSI score in participants with radiographic evidence of knee osteoarthritis was significantly higher in those with knee pain than in those without pain. The results of this study suggest that participants with significant low back pain and a higher number of painful sites are more susceptible to the influence of central sensitization. In addition, CSI score was higher in participants with low back pain and posterior lower leg pain than in those with low back pain alone, suggesting that the spread of pain may be due to central sensitization.


Borneol-based antisolvent-induced in situ forming matrix for crevicular pocket delivery of vancomycin hydrochloride.
Given its safety and apparent low aqueous solubility, borneol may serve as a matrix forming component of anti-solvent based in situ forming matrixes (ISMs) for crevicular pocket targeting. Drug-free and vancomycin hydrochloride-loaded borneol ISMs were evaluated for pH, density, viscosity, contact angle, surface tension, matrix formation, drug release behavior, in vitro degradability and antimicrobial activities. Density and pH values of borneol-based ISMs decreased with increasing borneol concentration. Given their markedly low viscosity could facilitate better injectability. The contact angles of the drug-free and vancomycin HCl-loaded borneol ISMs increased after being in contact with the agarose gel or the bulge tissue of porcine due to phase inversion. A dense borneol crystal matrix formed after using the highly concentrated ISM corresponded to fast matrix formation. The borneol-based ISM exhibited a sustainable drug release longer than 14 days with a diffusion-controlled release mechanism. Moreover, the developed ISM exhibited strong antimicrobial activities against various microbes. Thus, the vancomycin HCl-loaded borneol-based ISM is a potentially effective local anti-solvent-based ISM for periodontitis treatment via crevicular pocket injection.


Selection of Beauveria bassiana isolates to control Alphitobius diaperinus.
This paper presents results on isolates of the entomopathogenic fungus Beauveria bassiana for Alphitobius diaperinus (Coleoptera: Tenebrionidae) control. Of the 30 isolates tested, four (CG 71, CG 152, UNIOESTE 4, and UNIOESTE 40) resulted in mortality rates >or= 40% within 10 days. These mortality rates could be considered high because of the resistance of these species to B. bassiana. Tests were conducted using these isolates to estimate LC(50), mortality rate over time, vegetative growth, and conidial production on artificial medium and on insects. Isolates CG 71, CG 152, and UNIOESTE 4 showed the best performance and great potential to be used in an integrated management program in poultry farms to control A. diaperinus. Also, the molecular profiles of 12 isolates were analyzed using the RAPD technique. The high-virulence isolates presented a more homogeneous RAPD pattern than the others. Genetic sequencing of the ITS region was performed for one of the virulent isolates (UNIOESTE 4) and compared with sequences deposited at the NCBI database, confirming its taxonomical position as belonging to B. bassiana Clade A.


Extracranial-intracranial bypass surgery for intracranial aneurysm of the anterior cerebral circulation: A systematic review and meta-analysis.
The safety of extracranial-intracranial (EC-IC) bypass in the management of anterior circulation intracranial aneurysms (IAs) remains to be determined. This systematic review aims to summarize the existing evidence and provide guidance for the precise management of IAs. We constructed search strategies and comprehensively searched Pubmed, Medline, Embase, Web of science, and Cochrane library. This systematic review was actualized according to the PRISMA statement. We evaluated study quality using the methodological index for non-randomized study (MINORS). Effect sizes were pooled using a random-effects model. Heterogeneity between studies was assessed using the I 2 test. Publication bias was assessed using the Egger's test. The registration number for this systematic review is CRD42023396730. This systematic review included a total of 21 articles, involving 915 patients. Postoperative bypass patency rate was 99% (95% CI 0.98-1.00); short-term follow-up was 98% (95% CI 0.94-1.00); long-term follow-up was 95% (95% CI 0.93-0.97). The long-term follow-up occlusion rate of saphenous vein was higher than that of radial artery (OR 6.10 95% CI 1.04-35.59). Short-term surgery-related mortality was 0.3% (95% CI 0.000-0.012); long-term follow-up was 0.4% (95% CI 0.000-0.013); The proportion of patients with a score of 0-2 on the modified Rankin Scale (mRS) during long-term follow-up was 92% (95% CI 0.86-0.98). The incidence rates of long-term follow-up complications were: ischemic 3% (95% CI 0.01-0.06); hemorrhagic 1% (95% CI 0.00-0.03); neurological deficit 1% (95% CI 0.00-0.03); other 3% (95% CI 0.01-0.06). Most of the included studies were retrospective studies. Studies reporting preoperative status were not sufficient to demonstrate postoperative improvement. Lack of sufficient subgroup information such as aneurysm rupture status. EC-IC therapy for anterior circulation IAs has a high safety profile. Higher level of evidence is still needed to support clinical decision. https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42023396730, identifier: CRD42023396730.


Importance of method in the determination of syncytium-inducing phenotype of human immunodeficiency virus type 1 clinical isolates.
The in vitro syncytium induction capacity of human immunodeficiency virus type 1 (HIV1) isolates is an important marker in the progression of the disease. Two methods have been widely used to determine the biological phenotype of HIV1. These two methods, the direct MT-2 assay and the supernatant assay, were compared for the detection of syncytium-inducing (SI) variants on 275 blood samples obtained from 87 HIV infected patients during a 13 month follow-up period. A SI virus was detected in 152 blood samples. In 44 blood samples, the HIV isolate was found to be SI by only one method, but was SI by both methods in another blood sample of the follow up. Among SI carriers discordant results between the methods were more frequent when the patient was on antiretroviral therapy, and a transient reversion to a non syncytium-inducing (NSI) strain confirmed by both assays was sometimes observed. The supernatant assay has a 93% sensitivity and the direct MT-2 assay has a 78% sensitivity for detection of the SI phenotype. The supernatant assay is as rapid as and less tedious than the MT-2 assay. Antiretroviral therapy could have some effects in decreasing or even suppressing the SI part of the virus population of patients with SI phenotype.


Pressor response to tyramine after single 24-hour application of a selegiline transdermal system in healthy males.
Orally administered selegiline hydrochloride is a selective monoamine oxidase type B inhibitor at the recommended regimen of 10 mg/day, but it loses selectivity at higher doses. In bypassing first-pass metabolism, a 24-hour application of transdermally administered selegiline (7.8 mg/24 hr) yields fifty times greater systemic exposure than is provided by single oral doses. The current study was designed to demonstrate that, similar to the oral regimen, transdermally administered selegiline is devoid of the pressor effects associated with tyramine and classic monoamine oxidase type A inhibitors. A single-blind, staggered, parallel-group study of pressor response to tyramine during a single 24-hour application of one-quarter, one-half, or one selegiline transdermal system relative to baseline (drug-free) response to tyramine was conducted in three groups, each with five healthy male volunteers. The end point of pressor response was declared if a participant's systolic blood pressure rose by > 30 mmHg, heart rate decreased by > 25 bpm with an associated > 20-mmHg rise in systolic blood pressure, or a clinically significant change was observed in the electrocardiogram. Doses up to 600 mg were administered during the baseline phase and up to 200 mg during the active-treatment phase. Participants received escalating tyramine doses every 4 hours until the maximum or threshold dose was achieved. Doses up to 200 mg were tolerated without apparent increase in sensitivity in participants receiving one-quarter, one-half, or one selegiline transdermal system. All participants completed the trial, and no significant adverse events were reported. Monoamine oxidase type B inhibition was complete (100%) by 12 hours after initial application in all treatment groups while plasma levels of 3-methoxy-4-hydroxyphenylglycol (MHPG) after 24-hour application were unaffected relative to baseline. These results suggest that systemic selegiline levels may not predict the propensity for a hypertensive crisis associated with presumed nonselective doses and that the avoidance of peripheral monoamine oxidase type A inhibition in the gut via the selegiline transdermal system may provide a safe vehicle for administering selegiline at plasma levels beyond that which can be safely obtained after oral administration. These findings will need to be confirmed in a long-term dose setting.