Silibinin Reduces the Impact of Obesity on Invasive Liver Cancer.
Obesity is associated with non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). Obesity and metabolic abnormalities resulting in low-grade inflammation can increase the risk of developing NASH and HCC. NASH, a risk factor for HCC, is characterized by increased inflammation, lipid accumulation, and liver injury. Obesogenic proteins modulate signaling pathways that induce physiological changes including lipogenesis, ROS, and inflammation. Silibinin, a polyphenol in milk thistle seed, has been shown to have anti-inflammatory properties. Studies have yet to determine whether silibinin can be used to dissect the obesity-cancer link to delay progression of liver cancer. Using an in vitro model, sera from obese (OB), overweight (OW), or normal weight (NW) males (based on BMI) were used to determine the efficacy of silibinin to reduce the pro-tumorigenic properties of obesity. HepG2 cells were exposed to OB, OW, NW ± silibinin and tested for growth, ROS, lipogenesis, MMP-9, invasion and protein expression. Silibinin suppressed obesity-induced growth, ROS, lipogenesis, MMP-9, and cell invasion. These physiological changes corresponded with decreased FASN, IL-6, IL-1B, and phosphorylated Erk. We describe the differential effect of sera from OB, OW, and NW males on characteristics relevant for liver cancer and the potential use of silibinin to mitigate these effects.


Unique myological changes associated with ossified fabellae: a femorofabellar ligament and systematic review of the double-headed popliteus.
The fabella is a sesamoid bone embedded in the tendon of the lateral head of the gastrocnemius. It is the only bone in the human body to increase in prevalence in the last 100 years. As the fabella can serve as an origin/insertion for muscles, tendons, and/or ligaments (e.g., the oblique popliteal and fabellofibular ligaments), temporal changes in fabella prevalence could lead to temporal changes in "standard" knee anatomy. The aim of this study was to investigate unique myological changes to the posterolateral corner knee associated with ossified fabella presence and perform a systematic review to contextualize our results. Thirty-three fresh frozen cadaveric knees were considered. As the knees were all used for previous experimentation, the knees were in variable levels of preservation. Those with adequate preservation were used to determine ossified fabella presence/absence. When ossified fabellae were present, unique myologies associated with the fabella were recorded. A systematic review was performed on the double-headed popliteus to investigate possible correlations between this anatomical variant and the fabella. Of the 33 knees, 30 preserved enough soft tissue to determine fabella presence/absence: 16/30 knees had fabellae (five cartilaginous and 11 ossified). Eight of the eleven knees with ossified fabellae retained enough soft tissue to investigate the posterolateral knee anatomy. Of these, 4/8 exhibited unique myological changes. One knee had a double-headed popliteus muscle where one head originated from the medial side of a large, bulbous fabella. A systematic review revealed double-headed popliteus muscles are rare, but individuals are 3.7 times more likely to have a fabella if they have a double-headed popliteus. Another knee had a large, thick ligament stretching from the lateral edge of the fabella to the inferoposterior edge of the lateral femoral epicondyle, deep to the lateral collateral ligament (LCL) and near the popliteal sulcus. We found no mention of such a ligament in the literature and refer to it here as the "femorofabellar ligament". In all four knees, the plantaris and lateral gastrocnemius appeared to share a common tendinous origin, and the fabella was located at/near the junction of these muscles. In the case of the double-headed popliteus, the fabella clearly served as an origin for the plantaris. Despite being found in an average of 36.80% of human knees, most standard anatomical models fail to account for the fabella and/or the unique myological changes associated with fabella presence. Although our sample is small, these data highlight aspects of human biological variability generally not considered when creating generalized anatomical models. Further work is needed to identify additional changes associated with ossified fabellae and the functional consequences of omitting these changes from models.


Bioinspired NiO Nanospheres: Exploring In Vitro Toxicity Using Bm-17 and L. rohita Liver Cells, DNA Degradation, Docking, and Proposed Vacuolization Mechanism.
The present work demonstrated a novel Cleome simplicifolia-mediated green fabrication of nickel oxide nanoparticles (NiO NPs) to explore in vitro toxicity in Bm-17 and Labeo rohita liver cells. As-fabricated bioinspired NiO NPs were characterized by several analytical techniques. X-ray diffraction (XRD) revealed a crystalline face-centered-cubic structure. Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible diffuse reflectance spectroscopy (UV-DRS), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS) confirmed NiO formation. The chemical composition was confirmed by energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy. Brunauer-Emmett-Teller (BET) revealed the mesoporous nature. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed the formation of 97 nm diameter nanospheres formed due to the congregation of 10 nm size particles. Atomic force microscopy (AFM) revealed the nearly isotropic behavior of NiO NPs. Further, a molecular docking study was performed to explore their toxicity by binding with genetic molecules, and it was found that the docking energy was about -9.65284 kcal/mol. On evaluating the in vitro toxicity of NiO NPs for Bm-17 cells, the study showed that when cells were treated with a high concentration of NPs, cells were affected severely by toxicity, while at a lower concentration, cells were affected slightly. Further, on using 50 μg/mL, quick deaths of cells were observed due to the formation of more vacuoles in the cells. The DNA degradation study revealed that NiO NPs are significantly responsible for DNA degradation. For further confirmation, trypan blue assay was observed for cell viability, and morphological assessment was performed using inverted tissue culture microscopy. Further, the cytotoxicity of NiO NPs in L. rohita liver cells was studied. No toxicity was observed at 1 mg/L of NiO NPs; however, when the concentration was 30 and 90 mg/L, dark and shrank hepatic parenchyma was observed. Hence, the main cause of cell lysis is the increased vacuolization in the cells. Thus, the present study suggests that the cytotoxicity induced by NiO NPs could be used in anticancer drugs.


Rhabdomyolysis and drug abuse in a patient with bulimia nervosa.
Eating disorders, particularly bulimia nervosa, have long been associated with alcohol and illicit drug abuse. Impulsivity has been linked to substance abuse, but there are other explanations for this association. This comorbidity could be very harmful, even deadly, because of the medical consequences of drug abuse. This article reports on the case of a 23-year-old woman with bulimia nervosa, severe alcohol and illicit drug abuse, and impulsive behavior who developed rhabdomyolysis. One of the reasons for her drug addiction was the effect of these substances in appetite suppression. The patient successfully recovered after medical and psychological treatment. This article presents the case of a patient with bulimia nervosa and a serious medical complication of illicit drug abuse. It is important to be aware of the increased risk of morbidity and mortality for this patient group.


Remote-controlled magnetic pulmonary vein isolation using a new three-dimensional non-fluoroscopic navigation system: a single-centre prospective study.
Catheter ablation of atrial fibrillation (AF) focuses on pulmonary vein isolation (PVI), but the procedure is associated with significant X-ray exposure. Few data exist concerning the combination of remote magnetic navigation (RMN) and a new three-dimensional non-fluoroscopic navigation system (Carto(®) 3), which facilitates precise catheter navigation and limits X-ray exposure. To assess the efficacy and extent of fluoroscopic exposure associated with the combination of RMN and the Carto 3 system in patients requiring AF ablation. Between January and September 2011, catheter ablation was performed remotely using the Carto 3 system in 81 consecutive patients who underwent PVI for symptomatic drug-refractory AF. The radiofrequency generator was set to a fixed power ≤ 35W. The primary endpoint was wide-area circumferential PVI confirmed by spiral catheter recording during ablation and including additional lesion lines (left atrial roof and coronary sinus defragmentation) or complex fractionated atrial electrograms for persistent AF. Secondary endpoints included procedural data, complications and freedom from atrial tachycardia (AT)/AF. Mean age was 60 ± 9 years; 20% were women; 73% had symptomatic paroxysmal AF; 27% had persistent AF. The CHA2DS2-VASc score was 1.2 ± 1. Median procedure time was 3.5 ± 1 hours; median total X-ray exposure time was 13 ± 7 minutes; transseptal puncture and catheter positioning took 8 ± 4 minutes, left atrium electroanatomical reconstruction 1 ± 4 minutes and catheter ablation 3.5 ± 5minutes. Recurrences were AT (n=3; 3.7%), paroxysmal AF (n=8; 9.9%) and persistent AF (n=4; 4.9%); redo ablation was performed in these 15 (19%) patients. After a median follow-up of 15 ± 6 months and a single procedure, 71% of patients were free of symptoms; 84% remained asymptomatic after two procedures. RMN with irrigated catheters combined with the Carto 3 system can be effectively performed in patients requiring AF ablation with minimal use of fluoroscopy, but larger randomized studies are warranted.


Cytologic diagnosis of peripheral T-cell lymphoma manifesting as ascites. A case report.
Patients with malignant lymphoma seldom present with effusions without a known history of malignancy. Because of this, initial diagnosis of malignant lymphoma by effusion cytology is uncommon, with few reported cases. A 75-year-old male presented with fatigue, decreased appetite and progressively increasing abdominal girth over five weeks. Cytologic examination of ascitic fluid obtained by paracentesis revealed non-Hodgkin's lymphoma with a T-cell phenotype, confirmed by immunophenotypic and molecular studies. Approximately one week later, histologic examination of liver and bone marrow revealed involvement by lymphoma, demonstrating immunophenotypic and molecular profiles identical to those obtained from neoplastic lymphocytes recovered from the ascites fluid. This case demonstrates a rare presentation of peripheral T-cell lymphoma, clinically manifesting as ascites. In cases such as ours, where the effusion consists predominantly of small to intermediate-sized lymphocytes, distinguishing lymphoma from reactive lymphocytosis may be difficult. This case not only demonstrates the value of effusion cytology for lymphoma diagnosis but also illustrates how the use of various immunophenotypic and molecular techniques may assist the pathologist in properly diagnosing these difficult cases.


The teleambulance.
The teleambulance aims to facilitate adequate medical treatment by initiating and guaranteeing maximum preparation and immediate hospital handling of the patient either at the scene of an incident or during transportation. Following the problem definition and the identification of the medical fields the technology can address, field research was conducted to investigate the state of the art from the point of view of the prospective end user. This paper presents the development of the teleambulance workstation.


Cytochrome P450 genes induced by bispyribac-sodium treatment in a multiple-herbicide-resistant biotype of Echinochloa phyllopogon.
Incremental herbicide metabolism by cytochrome P450 monooxygenases (P450s) has been proposed as the basis for resistance to bispyribac-sodium (bispyribac) in a multiple-herbicide-resistant biotype of Echinochloa phyllopogon. Upon exposure to bispyribac, strong induction of bispyribac-metabolising P450 activity has been reported in the resistant line, indicating that P450s induced by bispyribac are involved in the bispyribac resistance. A polymerase chain reaction (PCR)-based cloning strategy was used to isolate 39 putative P450 genes from the bispyribac-resistant line of E. phyllopogon. Expression analysis by real-time PCR revealed that seven of the isolated genes were upregulated in response to bispyribac treatment of seedlings at the three-leaf stage. The transcript levels and protein sequences of the seven genes were compared between the bispyribac-resistant line and a susceptible line. CYP71AK2 and CYP72A254 were transcribed prominently in the bispyribac-resistant line. Amino acid polymorphisms were found in three genes, including CYP72A254. Upregulated expression of these genes is consistent with the inducible herbicide-metabolising P450 activity under bispyribac stress that was reported in a previous study. This is the first study to compare P450 genes in arable weed species in order to elucidate the mechanism for P450-mediated herbicide resistance.


[Effect of corticosteroids on tachyphylaxis induced by beta-2- receptor agonists in patients with bronchial asthma].
Fourteen patients with moderate bronchial asthma were selected for the study. Asthmatics were treated with intravenous salbutamol for one week. Peripheral blood lymphocyte beta-2-adrenergic receptor density and affinity were determined before and after the treatment. To investigate an effect of corticosteroids on beta-2-agonist induced desensitization of beta-2-receptors, beta-2-receptor density and affinity following prednisone administration have been determined in lymphocytes in 7 asthmatics. Patients have been given prednisone in the dose of 20 mg daily for 1 wk after salbutamol discontinuation. Spontaneous recovery of beta-2-adrenergic receptor density in the lymphocytes was estimated in further 7 asthmatics, not given corticosteroids following salbutamol therapy. Our data showed that down-regulation of beta-2-adrenergic receptors improves spontaneously and to significantly higher degree after corticosteroid therapy.


[Renal impairment induced by nonselective prostaglandin inhibitor].
The role of kidney function is of utmost importance in the maintenance of constant body fluid amount. However, kidneys are at a great risk of exposure due to high intrarenal blood flow and the existing concentrate system. Different damage can originate from renal clearance of various drugs, while metabolites or xenobiotics may be associated with potential nephrotoxic effects, particularly in the presence of additional risk factors. For analgesics such as nonsteroidal antirheumatics (NSARs), it is possible that they inhibit renal production of prostaglandins, potent vasodilators, resulting in tissue ischemia and direct toxicity. Therefore, the aim of this study was to evaluate changes in the cellular potential response of proximal tubules first in the presence of prostaglandin type E (PGE2) and then in the presence of Na-diclofenac, a nonselective prostaglandin inhibitor, and their effects on transport process of substrate (glucose), by using electrophysiological studies in experimental animals. PGE2 was found to play specific role at local intrarenal level, where they directly stimulated the processes of sodium and substrate (glucose) cotransport through luminal membrane of proximal tubules, without changing the control cell potential response. Results with diclofenac as a nonselective cyclooxygenase inhibitor and thus prostaglandin synthesis inhibitor revealed it to cause concentration-dependent depolarization of the cellular potential. It also reduces cotransport of the substrates and ions involved in intracellular equilibrium. At concentrations higher than therapeutic, diclofenac became nonspecific and therefore caused permanent depolarization. Accordingly, the results of these experimental studies may be useful to specialists treating patients suffering from renal dysfunction or kidney diseases. This can also contribute to rationalization of the therapeutic use of analgesics, greater caution and better recognition of the associated risk factors, in elderly or renal patients in particular.


Development of a Qualitative Quantitative Polymerase Chain Reaction Test to Identify Patients Failing First-Line Therapy to Non-Nucleotide Reverse Transcriptase Inhibitor.
Antiretroviral therapy (ART) can be compromised by selection of drug resistance strains, which can be promoted by lack of adherence during therapy and drug tolerance, and some of these drug-resistant strains can persist for years as minority populations. The K103N drug resistance mutation is selected by the use of non-nucleotide reverse transcriptase inhibitors, including nevirapine or efavirenz (EFV), used in low-income countries. Here we describe the use of a less expensive qualitative point mutation polymerase chain reaction (PMqPCRK103N) targeting K103N mutation. To validate the use of this methodology, we tested previously sequenced samples from patients treated with highly active ART with viral loads above 2,000 copies/ml and compared the results of our assay with Illumina deep sequencing. Due to its low cost and high specificity, this test is particularly suitable for low-income countries to screen for pretreatment resistance in patients either initiating ART or failing first-line regimens containing EFV.


Metagenomic assessment of body surface bacterial communities of the sea urchin, Tripneustes gratilla.
Sea urchins, including Tripneustes gratilla, are susceptible to a disease known as bald sea urchin disease, which has the potential to lead to economic losses in this emerging aquaculture industry in South Africa. This disease is characterized by lesions that form on sea urchin exoskeletal surfaces. This study aimed to characterize the body surface bacterial communities associated with T. gratilla, using a 16S rDNA gene metagenomics approach, to provide insight into the bacterial agents associated with this aquaculture species, as well as with this balding disease. Bacterial samples were collected from non-lesioned healthy animals obtained from natural locations along the eastern coast of South Africa, as well as from different cultured cohorts: non-lesioned healthy-, lesioned diseased- and non-lesioned stressed animals. A total of 1,067,515 individual bacterial operational taxonomic units (OTUs) were identified, belonging to 133 family-, 123 genus- and 113 species level OTU groups. Alpha diversity analyses, based on Chao1, Shannon and Simpson indices, showed that there were no statistically significant differences (ANOVA; P > 0.05) between the respective cohorts, as all cohorts displayed a high degree of bacterial diversity. Similarly, beta diversity analyses (Non-metric multidimensional scaling) showed a large degree of overlapping OTUs across the four cohorts. Within each cohort, various OTUs commonly associated with marine environments were found, predominantly belonging to the families Vibrionaceae, Saprospiraceae, Flavobacteriaceae and Sphingomonadaceae. Differential abundance analysis (DESeq2) revealed that OTUs that are differentially abundant across cohorts were likely not responsible for this balding disease, suggesting that complex bacterial agents, rather than a specific pathogenic agent, are likely causing this disease. Furthermore, the putative metabolic functions assigned to the bacterial communities showed that heterotrophic bacteria appear to be responsible for tissue lysis of degrading animal matter. The results from this study, obtained through univariate and multivariate-based approaches, contributes to future management strategies of this emerging aquaculture species by providing insight into the bacterial communities associated with both natural and cultured environments.


Induction of antiviral immunity by double-stranded RNA in a marine invertebrate.
Vertebrates mount a strong innate immune response against viruses, largely by activating the interferon system. Double-stranded RNA (dsRNA), a common intermediate formed during the life cycle of many viruses, is a potent trigger of this response. In contrast, no general inducible antiviral defense mechanism has been reported in any invertebrate. Here we show that dsRNA induces antiviral protection in the marine crustacean Litopenaeus vannamei. When treated with dsRNA, shrimp showed increased resistance to infection by two unrelated viruses, white spot syndrome virus and Taura syndrome virus. Induction of this antiviral state is independent of the sequence of the dsRNA used and therefore distinct from the sequence-specific dsRNA-mediated genetic interference phenomenon. This demonstrates for the first time that an invertebrate immune system, like its vertebrate counterparts, can recognize dsRNA as a virus-associated molecular pattern, resulting in the activation of an innate antiviral response.


Kinetic Studies on the Oxoiron(IV) Complex with Tetradentate Aminopyridine Ligand PDP*: Restoration of Catalytic Activity by Reduction with H2O2.
Oxoiron(IV) is a common catalytic byproduct observed in the oxidation of alkenes by the combination of H2O2 and nonheme iron catalysts including complex 1, FeIIPDP* (where PDP* = bis(3,5-dimethyl-4-methoxypyridyl-2-methyl)-(R,R)-2,2'-bipyrrolidine). The oxoiron(IV) species have been proposed to arise by O-O homolysis of the peroxyiron(III) or acylperoxyiron(III) intermediates formed during the presumed FeIII-FeV catalytic cycle and have generally been regarded as off-pathway. We generated complex 1IV═O (λmax = 730 nm, ε = 350 M-1 cm-1) directly from 1 and an oxygen atom donor IBXi-Pr (isopropyl 2-iodoxybenzoate) in acetonitrile in the temperature range from -35 to +25 °C under stopped-flow conditions. Species 1IV═O is metastable (half-life of 2.0 min at +25 °C), and its decay is accelerated in the presence of organic substrates such as thioanisole, alkenes, benzene, and cyclohexane. The reaction with cyclohexane-d12 is significantly slower (KIE = 4.9 ± 0.4), suggesting that a hydrogen atom transfer to 1IV═O is the rate limiting step. With benzene-d6, no significant isotope effect is observed (KIE = 1.0 ± 0.2), but UV-vis spectra show the concomitant formation of an intense 580 nm band likely due to the Fe(III)-phenolate chromophore, suggesting an electrophilic attack of 1IV═O on the aromatic system of benzene. Treatment of 1IV═O with H2O2 resulted in rapid decay of its 730 nm visible band (k = 102.6 ± 4.6 M-1 s-1 at -20 °C), most likely occurring by a hydrogen atom transfer from H2O2. In the presence of excess H2O2, the oxoiron(IV) is transformed into peroxyiron(III), as seen from the formation of a characteristic 550 nm visible band and geff = 2.22, 2.16, and 1.96 electron paramagnetic resonance (EPR) spectroscopy signals. Reductively formed 1III-OOH was able to re-enter the catalytic cycle of alkene epoxidation by H2O2, albeit with lower yields versus those of oxidatively formed (i.e., 1 + H2O2) peroxyiron(III) owing to a loss of ca. 40% active iron. As such, the oxoiron(IV) species can be reintroduced to the catalytic cycle with extra H2O2, acting as an iron reservoir. Alternatively, peroxycarboxylic acids, which have a stronger O-H bond dissociation energy, do not reduce 1IV═O, ensuring that more oxidant is productively employed in substrate oxidation. While this reaction with H2O2 may occur for other nonheme oxoiron(IV) complexes, the only previously reported examples are 3IV═O and 4IV═O, which are reduced by hydrogen peroxide 130- and 2900-fold more slowy, respectively (as in Angew. Chemie - Int. Ed. 2012 , 51 ( 22 ), 5376 - 5380 , DOI: 10.1002/anie.201200901 ).


Antiviral modes of action of the novel compound GLY-15 containing pyrimidine heterocycle and moroxydine skeleton against tobacco mosaic virus.
Plant virus diseases are difficult to prevent and control, causing serious economic losses to the agricultural production world. To develop new pesticides with antiviral activity, a serial of compounds containing the structure of pyrimidine and moroxydine were synthesized, among which GLY-15 exhibited good antiviral activity against tobacco mosaic virus (TMV), while the mechanism of antiviral activity remains to be clarified. GLY-15 treatment significantly inhibited the formation of necrotic spots caused by TMV in Nicotiana glutinosa, and effectively suppressed the systemic transportation of TMV expressing a reporter gene (p35S-30B:GFP) in N. benthamiana and markedly reduced the accumulation of a movement deficient TMV in plants as well as viral RNA accumulation in tobacco protoplasts. The results of RNA sequencing showed that GLY-15 induced significant differential expression of genes or pathways involved in the stress response, defense response and signal transduction, phytohormone response and metabolism. Among them, real-time quantitative PCR validated that the expression of 12 critical genes such as heat shock protein, receptor kinase, cell-wall-related protein, disease-related protein and glucan endo-1,3-β-glucosidase were significantly up-regulated. In addition, GLY-15 triggered reactive oxygen species (ROS) production and induced the activity of several crucial defense related enzymes in plants. The results of molecular docking showed potential binding ability of GLY-15 with TMV helicase and the coat protein. This study provide valuable insights into antiviral mechanism of action for GLY-15, which is expected to be applied as a pesticide for the management of plant viruses. © 2022 Society of Chemical Industry.


Tyrosinase gene mutations associated with type IB ("yellow") oculocutaneous albinism.
We have identified three different tyrosinase gene mutant alleles in four unrelated patients with type IB ("yellow") oculocutaneous albinism (OCA) and thus have demonstrated that type IB OCA is allelic to type IA (tyrosinase negative) OCA. In an inbred Amish kindred, type IB OCA results from homozygosity for a Pro----Leu substitution at codon 406. In the second family, type IB OCA results from compound heterozygosity for a type IA OCA allele (codon 81 Pro----Leu) and a novel type IB allele (codon 275 Val----Phe). In the third patient, type IB OCA results from compound heterozygosity for the same type IB allele (codon 275 Val----Phe) and a novel type IB OCA allele. In a fourth patient, type IB OCA results from compound heterozygosity for the codon 81 type IA OCA allele and a type IB allele that contains no identifiable abnormalities; dysfunction of this type IB allele apparently results from a mutation either well within one of the large introns or at some distance from the tyrosinase gene. In vitro expression of the Amish type IB allele in nonpigmented HeLa cells demonstrates that the Pro----Leu substitution at codon 406 greatly reduces but does not abolish tyrosinase enzymatic activity, a finding consistent with the clinical phenotype.


Identification of colorectal cancer-associated macrophage biomarkers by integrated bioinformatic analysis.
The aim of this study was to explore colorectal tumor-associated macrophage (TAM) biomarkers for early diagnosis and surveillance of colorectal cancer (CRC). We used bioinformatic methods to screen array expression data of CRC-related macrophages (GEO: GSE29214) to detect the differentially expressed genes (DEGs) between CRC-related macrophages and normal control cells. We found 431 DEGs in TAMs compared with the control group; 399 were up-regulated and 32 were down-regulated. A functional enrichment analysis showed that the DEGs were involved in positive regulation of the ERK1 and ERK2 cascade, cell activation involved in the immune response, cytokine-mediated signaling pathway, and receptor activity, all of which were significantly enriched. We constructed a protein-protein interaction (PPI) network to identify hub genes. We identified 10 hub genes: ITGB2, ITGAM, C3AR1, PTAFR, C3, CYBB, FCER1G, PLAU, STOM, and GPR84, in the PPI network. We verified the results using array expression data of peripheral blood mononuclear cells (GEO: GSE47756). The results showed that the expression trends of CYBB, PLAU, and STOM were consistent with those found in the GSE29214 dataset. Further verification with The Cancer Genome Atlas and Human Protein Atlas showed that the high expression of PLAU in TAMs was statistically significant (P<0.05). We concluded that PLAU may be a biomarker of CRC-associated macrophages and may have prognostic and predictive significance for clinical utility in CRC management.


Fecal Microbiota Transplantation for Recurrent Clostridium difficile Infection Reduces Recurrent Urinary Tract Infection Frequency.
Broad-spectrum antibiotics for recurrent multidrug-resistant urinary tract infections (UTIs) disrupt the gut microbiome and promote antibiotic resistance. Fecal microbiota transplantation led to resolution of recurrent Clostridium difficile, significantly decreased recurrent UTI frequency, and improved antibiotic susceptibility profile of UTI-causing organisms.


Hybrid tungsten-carbon 2D nanostructures viain situgasification of carbon substrates driven by ebeam irradiation of WO2.9microparticles.
Since the transmission electron microscope (TEM) has the capacity to observe the atomic structure of materials,in situTEM synthesis methods are uniquely suited to advance our fundamental understanding of the bottom-up dynamics that drive the formation of nanostructures. E-beam induced fragmentation (potentially identified as a manifestation of Coulomb explosion) and electron stimulated desorption are phenomena that have received attention because they trigger chemical and physical reactions that can lead to the production of various nanostructures. Here we report a simple TEM protocol implemented on WO2.9microparticles supported on thin amorphous carbon substrates. The method produces various nanostructures such as WC nanoparticles, WC supported films and others. Nevertheless, we focus on the gradual graphitization and gasification of the C substrate as it interacts with the material expelled from the WO2.9microparticles. The progressive gasification transforms the substrate from amorphous C down to hybrid graphitic nanoribbons incorporating W nanoparticles. We think these observations open interesting possibilities for the synthesis of 2D nanomaterials in the TEM.


[The progress of studying the mechanisms of immune cells in the regulation of non-alcoholic fatty liver diseases].
Non-alcoholic fatty liver diseases (NAFLD) are liver diseases which are strongly associated with obesity. The incidence of NAFLD has been gradually increased in recent years. According to different pathology of the liver, NAFLD include three disease states: simple fatty liver, hepatitis (nonalcoholic steatohepatitis, NASH) and fatty liver cirrhosis. The accumulations of fat around the internal organs, which secrete a wide range of adipokines, such as interleukin-6, contribute the development and progress of NAFLD in obesity. In addition, the liver has been recognized as an immune organ containing several immune cells, for instance, inherited macrophages in the live (kupffer cells), nature killer cells, T lymphocytes, T regulatory cells (Tregs), myeloid-derived suppressor cells (MDSCs), and dendritic cells. These immune cells secret a number of cytokines, such as tumor necrosis factor (TNF-α) and monocyte chemoattractant protein (MCP-1), play important roles in the development of NAFLD. Moreover, a high level of fatty acids in the liver microenvironment, leads to suppress CD4(+)T cells and the loss of CD4(+)T cells in the microenvironment of the liver may be one of key factors which is associated with the progress of NAFLD to cirrhosis and liver cancer. This article aims to review the recent studies in the mechanisms of these immune cells involving in the progress of NAFLD, and to explore the novel strategies targeting these immune cells to control the prevalence of NAFLD and reduce liver fibrosis.


A Combination of Anti-PD-L1 Treatment and Therapeutic Vaccination Facilitates Improved Retroviral Clearance via Reactivation of Highly Exhausted T Cells.
PD-1-targeted therapies have shown modest antiviral effects in preclinical models of chronic viral infection. Thus, novel therapy protocols are necessary to enhance T cell immunity and viral control to overcome T cell dysfunction and immunosuppression. Here, we demonstrate that nanoparticle-based therapeutic vaccination improved PD-1-targeted therapy during chronic infection with Friend retrovirus (FV). Prevention of inhibitory signals by blocking PD-L1 in combination with therapeutic vaccination with nanoparticles containing the microbial compound CpG and a CD8+ T cell Gag epitope peptide synergistically enhanced functional virus-specific CD8+ T cell responses and improved viral clearance. We characterized the CD8+ T cell populations that were affected by this combination therapy, demonstrating that new effector cells were generated and that exhausted CD8+ T cells were reactivated at the same time. While CD8+ T cells with high PD-1 (PD-1hi) expression turned into a large population of granzyme B-expressing CD8+ T cells after combination therapy, CXCR5-expressing follicular cytotoxic CD8+ T cells also expanded to a high degree. Thus, our study describes a very efficient approach to enhance virus control and may help us to understand the mechanisms of combination immunotherapy reactivating CD8+ T cell immunity. A better understanding of CD8+ T cell immunity during combination therapy will be important for developing efficient checkpoint therapies against chronic viral infections and cancer.IMPORTANCE Despite significant efforts, vaccines are not yet available for every infectious pathogen, and the search for a protective approach to prevent the establishment of chronic infections, i.e., with HIV, continues. Immune checkpoint therapies targeting inhibitory receptors, such as PD-1, have shown impressive results against solid tumors. However, immune checkpoint therapies have not yet been licensed to treat chronic viral infections, since a blockade of inhibitory receptors alone provides only limited benefit, as demonstrated in preclinical models of chronic viral infection. Thus, there is a high interest in the development of potent combination immunotherapies. Here, we tested whether the combination of a PD-L1 blockade and therapeutic vaccination with functionalized nanoparticles is a potent therapy during chronic Friend retrovirus infection. We demonstrate that the combination therapy induced a synergistic reinvigoration of the exhausted virus-specific CD8+ T cell immunity. Taken together, our results provide further information on how to improve PD-1-targeted therapies during chronic viral infection and cancer.


Fatal Rhabdomyolysis in a COVID-19 Patient on Rosuvastatin.
It is well-established by now that COVID-19 can have a wide variety of neuromuscular manifestations, including rhabdomyolysis. Weakness and elevated creatinine kinase (CK) have been documented as the initial presentation of COVID-19. Myopathy from statin use has also been well-established since the introduction of this class of medication, and the common pathologic mechanism of both entities may have been mitochondrial dysfunction. We present here the case of a COVID-19 patient on rosuvastatin who developed rhabdomyolysis with CK above 1,000,000 units/L. The patient did not present with any respiratory difficulty and responded poorly to treatment, resulting in his untimely demise. COVID-19 may have accentuated an otherwise survivable condition by means of extra stress on mitochondrial homeostasis. Understanding the actual mechanism will be important in the development and utilization of medications in the fight against COVID-19.


Effects of the individual isomers cis-9,trans-11 vs. trans-10,cis-12 of conjugated linoleic acid (CLA) on inflammation parameters in moderately overweight subjects with LDL-phenotype B.
Immune-modulating effects of CLA have been reported in animals, but results are inconsistent. In humans, CLA has shown no effects or only minor effects on immune function. The objective of this study was to evaluate the immune-modulating effects of 3 g cis-9,trans-11 (c9,t11) vs. trans-10,cis-12 (t10,c12) CLA isomers in a population with a high risk of coronary heart disease characterized by moderate overweight (body-mass index, 25-32.5 kg/m2) in combination with LDL-phenotype B (> or = 35% small LDL cholesterol, density > or = 1.040 g/mL). After a run-in period of 1 wk, 42 men and women were randomly allocated to the c9,t11 CLA group, the t10,c12 CLA group, or the placebo group. Effects of 13 wk of consumption of 3 g of CLA isomers on cytokine production by ex vivo lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMC) and whole blood, and on plasma C-reactive protein (CRP) concentrations were evaluated. To generate hypotheses for future studies, protein expression patterns of 42 cytokines, chemokines, and growth factors were evaluated with an antibody array in pooled, nonstimulated, fasting plasma samples. LPS induced interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha production by PBMC, and whole blood as well as plasma CRP concentrations were not significantly changed by the c9,t11 and the t10,c12 CLA isomers. The cytokine expression profile in nonstimulated plasma suggested that both CLA isomers induced a specific inflammatory signature, in which the c9,t11 CLA group showed more activity in terms of numbers of proteins regulated. We conclude that daily consumption of 3 g of c9,t11 or t10,c12 CLA isomer did not affect LPS-stimulated cytokine production by PBMC or whole blood and plasma CRP levels. Inflammatory signatures in fasting, nonstimulated plasma as determined by an antibody array may indicate enhanced immune function by both CLA isomers.


The Plastome Sequences of Triticum sphaerococcum (ABD) and Triticum turgidum subsp. durum (AB) Exhibit Evolutionary Changes, Structural Characterization, Comparative Analysis, Phylogenomics and Time Divergence.
The mechanism and course of Triticum plastome evolution is currently unknown; thus, it remains unclear how Triticum plastomes evolved during recent polyploidization. Here, we report the complete plastomes of two polyploid wheat species, Triticum sphaerococcum (AABBDD) and Triticum turgidum subsp. durum (AABB), and compare them with 19 available and complete Triticum plastomes to create the first map of genomic structural variation. Both T. sphaerococcum and T. turgidum subsp. durum plastomes were found to have a quadripartite structure, with plastome lengths of 134,531 bp and 134,015 bp, respectively. Furthermore, diploid (AA), tetraploid (AB, AG) and hexaploid (ABD, AGAm) Triticum species plastomes displayed a conserved gene content and commonly harbored an identical set of annotated unique genes. Overall, there was a positive correlation between the number of repeats and plastome size. In all plastomes, the number of tandem repeats was higher than the number of palindromic and forward repeats. We constructed a Triticum phylogeny based on the complete plastomes and 42 shared genes from 71 plastomes. We estimated the divergence of Hordeum vulgare from wheat around 11.04-11.9 million years ago (mya) using a well-resolved plastome tree. Similarly, Sitopsis species diverged 2.8-2.9 mya before Triticum urartu (AA) and Triticum monococcum (AA). Aegilops speltoides was shown to be the maternal donor of polyploid wheat genomes and diverged ~0.2-0.9 mya. The phylogeny and divergence time estimates presented here can act as a reference framework for future studies of Triticum evolution.


[Anemia and chronic kidney failure].
Anaemia is a common manifestation ofa chronic kidney failure. It is caused by a relative shortage oferythropoetine (EPO) and iron deficite with its metabolism defect. The most important factor in the pathogenesis of iron metabolism defects is hepcidin. Hepcidin maintains the iron homeostasis in the organism. The therapy of renal anaemia is based on the iron substitution and erythropoiesis stimulating agents (ESA) application. The most common reasons for the resistance to ESA are (after iron deficiency) inflammation and malnutrition.


[Changes in plasma inner ear hormones after endolymphatic sac drainage and steroid-instillation surgery (EDSS)].
We treated 33 cases of intractable Meniere's disease with endolymphatic sac drainage and steroid-instillation surgery (EDSS), attaining good long-term results in vertigo and hearing. To elucidate how EDSS affects the diseased inner ear, we examined changes in plasma inner ear hormones after EDSS. Among inner ear hormones, plasma vasopressin was significantly decreased after EDSS compared to after mastoidectomy. In cases with good long-term results in vertigo and hearing, postoperative plasma vasopressin remained lower over the long term than in cases with poor results. In cases with negative glycerol test results one year after surgery, postoperative plasma vasopressin also remained significantly lower over the long term than in cases with positive results. Previous studies reported that vestibular neurons projected into hypothalamic supraoptic and paraventricular nuclei and that changes in the inner ear pressure were related to plasma vasopressin. Taken together with present findings, this suggests that EDSS may reduce plasma vasopressin through modification of the diseased inner ear environment, resulting in improved inner ear function.


The Impact of Cooperative Learning on University Students' Academic Goals.
Cooperative learning encourages the development of interpersonal skills and motivates students to participate more actively in the teaching and learning process. This study explores the impact of cooperative learning on the academic goals influencing university students' behavior and leading to the attainment of a series of academic objectives. To this end, a quasi-experimental pretest-posttest control group design was used, with a sample of 509 university students from Preschool, Primary and Social Education undergraduate degree courses. Using the Academic Goals Questionnaire (AGQ), pretest and posttest measures were taken via self-reports to evaluate three types of academic goals: learning goals, social reinforcement goals and achievement goals. The results show that cooperative learning is an effective tool for encouraging university students to develop academic goals that motivate them to fully engage with the tasks they are set in order to acquire knowledge and skills (learning goals). In addition, when students are asked to work as part of a team on an autonomous basis without the structure and supervision necessary to ensure a minimum standard of cooperation, they display a greater tendency toward social reinforcement goals than toward learning and achievement goals. These findings contribute new knowledge to the conceptual framework on cooperative learning. Goals may be considered one of the most important variables influencing students' learning and the use of cooperative learning techniques in university classrooms creates the necessary conditions for encouraging students to develop goals oriented toward learning.


Thigh pain in cementless total hip arthroplasty. A comparison of two systems at 2 years' follow-up.
Cementless total hip arthroplasties using 110 porous-coated anatomic (PCA) and 105 consecutive Mallory-Head (MH) prostheses were reviewed using prospectively gathered clinical data and a detailed radiographic analysis. In an earlier experience with the PCA, 13% of patients experienced thigh pain at 1 year and 23% at 2 years. Positive radiographic correlates with thigh pain included subsidence, distal cortical hypertrophy, a tight diaphyseal fit, and stem tip sclerosis. In the current MH study group, the incidence of thigh pain was 7% at 1 year and 3% at 2 years. No radiographic findings correlated positively with thigh pain. The results of this short-term clinical comparison suggest that a long, straight, tapered design offered better initial stability than its anatomic counterpart as reflected in the much lower incidence of thigh pain. These results may translate into improved osseous integration and long-term success.


The Mediating Role of Placental Weight Change in the Association Between Prenatal Exposure to Thallium and Birth Weight: A Prospective Birth Cohort Study.
Background: Previous studies have demonstrated the embryotoxicity and fetotoxicity of thallium (Tl). However, the effects of prenatal exposure to Tl on birth weight and placental weight and the mediating role of placental weight in the association of Tl with birth weight remain unclear. Methods: We recruited 2,748 participants from the ongoing Prenatal Environment and Offspring Health Cohort (PEOH Cohort) study, which was initiated in 2016 in Guangzhou, China. The Tl concentrations in maternal urine samples collected during the first and third trimester were determined by inductively coupled plasma mass spectrometry. Birth weight and placental weight were extracted from maternal medical records. Results: Pregnant women exposed to the highest tertile of Tl in the first trimester (β = -42.7 g, 95% CI: -82.3, -3.1 g) and third trimester (β = -50.6 g, 95% CI: -99.0, -2.3 g) had babies with lower birth weights than those exposed to the lowest tertile. We also found significant negative associations of exposure to Tl concentrations in the first and third trimester with placental weight. Mediation analyses showed that 50.3% (95% CI: 15.9, 79.2%) and 33.5% (95% CI: 1.3, 80.3%) of the effects of Tl exposure in the first and third trimester on birth weight were mediated by decreased placental weight. Conclusion: Our results suggest that prenatal exposure to Tl is negatively associated with birth weight and that this association may be mediated by decreased placental weight.


Novel Filler-Filled-Type Polymer Electrolyte Membrane for PEFC Employing Poly(vinylphosphonic acid)-b-polystyrene-Coated Cellulose Nanocrystals as a Filler.
Low-acidity polymer electrolyte membranes are essential to polymer electrolyte fuel cells (PEFCs) and water electrolysis systems, both of which are expected to be next-generation energy and hydrogen sources. We developed a new type of high-performance polymer electrolyte membrane (PEM) in which the core particles are precisely electrolyte polymer coated and filled into binder resin. Cellulose nanocrystals (CNCs), which have attracted attention as light, rigid, and sustainable materials, were selected as the core material for the filler. The CNC surface was coated with a new block copolymer containing a proton conductive polymer of poly(vinylphosphonic acid) (PVPA) and a hydrophobic polymer of polystyrene (PS) using RAFT polymerization with particles (PwP) we developed. The pelletized fillers and the filler-filled polycarbonate membranes achieved proton conductivities of over 10-2 S/cm with lower activation energies and much weaker acidity than the Nafion membrane.


Microinjecting holo-aequorin into dechorionated and intact zebrafish embryos.
The injection of holo-aequorin into embryos at the one-cell stage, along with the use of a simple photomultiplier tube or luminescence imaging system, allows transient localized elevations of free cytosolic Ca(2+) to be recorded and observed during the first 24 h of zebrafish development. The technique for loading dechorionated or intact one-cell stage zebrafish embryos with holo-aequorin is described here.


Development of new brain metastases in triple negative breast cancer.
Brain metastases are common in patients with breast cancer, and those with triple negative status have an even higher risk. Triple negative status is currently not considered when managing brain metastases. To determine whether triple negative breast cancer (TNBC) patients with brain metastases have a higher burden of intracranial disease and whether WBRT has a survival benefit in this cohort of patients. We conducted a retrospective cohort study with 85 patients meeting the inclusion criteria. 25% of patients had TNBC. 95% of the patients in this study received SRS and 48% received WBRT. The average number of new brain metastases from time of initial brain imaging to radiation therapy was 0.67 ± 1.1 in the non-TNBC status patients and 2.6 ± 3.7 in the triple negative status patients (p = 0.001). A cox proportional hazards model showed that WBRT does not significantly affect overall survival in patients with TNBC (HR 1.48; 95% CI 0.47-4.67; p = 0.50). Our findings highlight the highly aggressive intracranial nature of TNBC. The rate of new brain metastasis formation is higher in TNBC patients compared to non-TNBC patients. Furthermore, there is no survival benefit for WBRT in TNBC patients. These findings are relevant for clinicians planning brain radiation for TNBC patients as they may find more brain metastases at the time of brain radiation than they anticipated based on initial brain imaging.


Surgical treatment of slipped capital femoral epiphysis (SCFE) by Dunn procedure modified by Ganz: a systematic review.
Treatment of SCFE is still controversial, especially in moderate and severe forms. Dunn osteotomy performed with the Ganz approach became very popular in the last decade, although it is a complicated and challenging surgical procedure with a risk of AVN. The aim of our study was to analyze the current literature verifying the effectiveness of this surgical procedure, with specific attention to the incidence of AVN and other complications. A systematic review on the subject was performed according to the PRISMA guidelines. A literature search was performed by searching all published articles about the topic in the databases. The articles were screened for the presence of the following inclusion criteria: patients affected by slipped capital femoral epiphysis (SCFE) surgically treated by Dunn osteotomy using the Ganz surgical approach. All the patients affected by pathologies other than SCFE, treated without surgery or with procedures not including a surgical hip dislocation were excluded. Based on inclusion and exclusion criteria, 23 studies were included in our systematic review. Selected articles were published from 2009 to 2021 and they included 636 overall hips. According to the selected articles, Dunn osteotomy modified by Ganz, performed by an experienced surgeon, allows for anatomical reduction of moderate or severe SCFE with a low incidence of AVN. The few papers with long term follow-up, reported no progression of hip osteoarthritis, however, since the patients are adolescent at surgery, longer follow-up studies are needed to validate this statement. It is still debated if better results are obtained in stable or unstable SCFE. The indication of this procedure in mild SCFE remains controversial. 3.


Comparison of the inhibition potential of parthenolide and micheliolide on various UDP-glucuronosyltransferase isoforms.
Parthenolide (PTL) and micheliolide (MCL) are sesquiterpene lactones with similar structures, and both of them have been reported to exhibit multiple biochemical and pharmacological activities. This study aims to investigate the inhibition of these two compounds on the activity of UDP-glucuronosyltransferases (UGTs). In vitro incubation mixture for recombinant UGTs-catalyzed glucuronidation metabolism of 4-methylumbelliferone (4-MU) was utilized to investigate the inhibition potential. Inhibition kinetics (including inhibition type and parameters) were determined, and in silico docking was employed to elucidate the inhibition difference between PTL and MCL on UGT1A1. MCL showed no inhibition toward all the UGT isoforms, and PTL showed strong inhibition toward UGT1A1. The half-maximal inhibitory concentration (IC50) of PTL on the activity of UGT1A1 was determined to be 64.4 μM. Inhibition kinetics determination showed that PTL exerted noncompetitive inhibition toward UGT1A1, and the inhibition kinetic constant (Ki) was determined to be 12.1 μM. In silico docking method has been employed to show that hydrogen bonds between PTL and the activity cavity of UGT1A1 contributed to the stronger inhibition of PTL on the activity of UGT1A1 than MCL. In conclusion, PTL can more easily induce drug-drug interaction (DDI) with clinical drugs mainly undergoing UGT1A1-catalyzed glucuronidation.


Inflammatory diseases of the nail unit.
inflammatory disorders of the nail unit are frequently encountered in clinical medicine and are a cause of significant morbidity. Psoriasis, lichen planus, chronic paronychia, and trachyonychia will be discussed. An approach to diagnosis and management of these disorders requires knowledge of nail unit anatomy, consideration of associated systemic manifestations, and patient education with respect to prognosis and management of factors that will maximize disease improvement. The nail unit responds with a limited number of signs to disparate clinical entities such that there is some overlap in clinical presentation between inflammatory conditions. Nail unit biopsy may therefore be useful in establishing a specific diagnosis.


Building a relationship: communications and relationships between staff and stroke patients on a rehabilitation ward.
Communications among staff and patients on a stroke rehabilitation ward form the focus of this article, which reports on some aspects of a larger study using a grounded theory approach. Tape-recorded interviews were transcribed and analysed concurrently according to recommendations for the approach. A main theme entitled building a relationship was identified, and this process was found to occur in a context varying from participative at one end of a continuum to hierarchical at the other. Building a relationship was found to be influenced by role, personal qualities and organizational context. Appropriate relationships between role-holders were subject to negotiation, leading to a resulting congruence or incongruence between participants' expectations of each other and their roles. Personal qualities were brought into play in the process, with patients' views of staff and staff views of patients both being influential. Some of these views seemed to parallel what has been described in earlier literature as 'the sick role' and the labelling of patients as 'good' or 'bad'. Responses to personal qualities led to nurses ascribing meaning to patients' behaviour in terms of adjustment to their stroke, giving time to them to help them to adjust, and withdrawal and handing over to other staff if this strategy failed. Organizational context also had an influence on building a relationship, with time constraints being identified particularly by nurses, and the need to fit in the most essential aspects of care. Place was also important, in that nurses were confined to the ward as a work location, whereas other therapists and doctors worked in other places and sometimes had the facility to take patients off the ward to concentrate on therapy. The findings are discussed against the background of related literature and the conclusion is drawn that the crucial role of nurses in rehabilitation is not recognized and valued, and that shortages of resources-especially suitably qualified and trained nursing staff-are a negative influence on building the relationships which are vital to successful rehabilitation.


Update on road traffic crashes: progress in the Middle East.
Road traffic injuries comprise the major share of all injuries globally. Traffic injuries kill 1.2 million people annually and injure 40 times as many, leaving a subsequent number totally disabled. Globally we spend approximately US $500 billion annually. The Middle East encompasses West Asia and North Africa and is very diverse economically, culturally and socially. Prevention and management of road traffic crashes and injuries is difficult. Comparative data are not readily available and therefore developing unified policies is a mammoth task. Implementation of best practices is not uniformly advocated due to socioeconomic and cultural differences. Enforcement of endorsed legislation on road traffic safety is not uniform in the region. Professional staff to combat this pandemic are scarce and it is important that capacity building, knowledge sharing, and increased political will becomes a priority in the region. This paper discusses the problems encountered in the prevention and management of road traffic injuries from the site of injury to rehabilitation and social reintegration. The role of Oman and that of the Bone and Joint Decade in the United Nations on Global Road Safety and its update is highlighted.


Phospholipid fatty acids in cord blood: family history and development of allergy.
The fatty acid composition of umbilical cord serum phospholipids was investigated by gas chromatography in 33 infants with allergic and 35 babies of non-allergic mothers. The relative levels of the linoleic acid metabolites C20:3, arachidonic acid (AA, C20:4) and C22:4n-6, and two alpha-linolenic acid metabolites, i.e. eicosapentaenoic acid (EPA, 20:5) and docosahexaenoic acid (DHA, C22:6) were significantly higher in infants of allergic mothers than in non-allergic mothers (all p < 0.05). Furthermore, an altered proportional relationship between the various fatty acids in n-6 series fatty acids and between n-3 and n-6 series fatty acids was present already at birth in infants who developed allergic disease during their first 6 years of life. These observations cannot be employed for the prediction of allergy, however, as the individual variations were considerable.


Triggers for autism: genetic and environmental factors.
This report reviews the research on the factors that cause autism. In several studies, these factors have been verified by reproducing them in autistic animal models. Clinical research has demonstrated that genetic and environmental factors play a major role in the development of autism. However, most cases are idiopathic, and no single factor can explain the trends in the pathology and prevalence of autism. At the time of this writing, autism is viewed more as a multi-factorial disorder. However, the existence of an unknown factor that may be common in all autistic cases cannot be ruled out. It is hoped that future biological studies of autism will help construct a new theory that can interpret the pathology of autism in a coherent manner. To achieve this, large-scale epidemiological research is essential.


Distribution of Micropogonias furnieri (Pisces: Sciaenidae) in Sepetiba Bay, Rio de Janeiro, Brazil.
Distribution of Micropogonias furnieri in the Sepetiba Bay, an ecosystem highly important as rearing ground for this species that represents 5.8% of the numerical catches by otter trawling, was analyzed based in monthly and bi-monthly samplings from two programmes (beach seines = continental margin; and otter trawl = inside of the Bay), over three annual cycles, between 1993 and 1997. Spatial comparisons were assessed by dividing the Bay in two zones for the beach seines (inner and outer Bay) and three zones for the otter trawl (inner, central and outer Bay), following depth, salinity and transparency gradient, as well as for the influence of the sea. Beach seines (30 m extension, a 10 m length x 2.5 m of height x 7 mm mesh) and otter trawls were used. Trawls covered 1.5 km and the boat (net mouth 8 m, mesh size 12 mm between opposites knots in the cod). Temperature (degree C) and salinity (p.s.u.) were taken in each sampling; depth (m) and transparency (m) were measured during the trawl. Fish from beach seine were basically young-of-the-year; in the trawl they were individuals of larger size with total length varying from 70 mm to 300 mm. Spatially, highest CPUEs were found for the continental margin in the inner Bay, and for the inside Bay, in the inner and central zones. Highly significant correlations were detected among M. furnieri abundance and low salinity, transparency and depth in the inner Bay, with no defined pattern for the continental margin. Temporally, differences in fish abundance were shown only for the second annual cycle (1996) for the continental margin, with peaks in September/October.


Persistent adrenergic reinnervation of previously denervated muscle in cat.
Synapse formation by aberrant nerves at chronically denervated intrinsic laryngeal muscles was observed even at 21 months after the transection of the recurrent laryngeal nerve in cats. Some of the aberrant nerves were adrenergic, and these nerve terminals were in synaptic contact with the postsynaptic membrane of the neuromuscular junctions together with non-adrenergic nerve terminals. Therefore, heterogeneous reinnervation between adrenergic nerve and muscle was found to be maintained for a long time.


Vacuolar cation/H+ antiporters of Saccharomyces cerevisiae.
We previously demonstrated that Saccharomyces cerevisiae vnx1Δ mutant strains displayed an almost total loss of Na(+) and K(+)/H(+) antiporter activity in a vacuole-enriched fraction. However, using different in vitro transport conditions, we were able to reveal additional K(+)/H(+) antiporter activity. By disrupting genes encoding transporters potentially involved in the vnx1 mutant strain, we determined that Vcx1p is responsible for this activity. This result was further confirmed by complementation of the vnx1Δvcx1Δ nhx1Δ triple mutant with Vcx1p and its inactivated mutant Vcx1p-H303A. Like the Ca(2+)/H(+) antiporter activity catalyzed by Vcx1p, the K(+)/H(+) antiporter activity was strongly inhibited by Cd(2+) and to a lesser extend by Zn(2+). Unlike as previously observed for NHX1 or VNX1, VCX1 overexpression only marginally improved the growth of yeast strain AXT3 in the presence of high concentrations of K(+) and had no effect on hygromycin sensitivity. Subcellular localization showed that Vcx1p and Vnx1p are targeted to the vacuolar membrane, whereas Nhx1p is targeted to prevacuoles. The relative importance of Nhx1p, Vnx1p, and Vcx1p in the vacuolar accumulation of monovalent cations will be discussed.


Human alpha 1 (XIII) collagen gene. Multiple forms of the gene transcripts are generated through complex alternative splicing of several short exons.
The structure of the gene for the human alpha 1(XIII) collagen chain (COL13A1) was determined from genomic clones spanning 140,000 base pairs (bp), including about 3,000 bp of the 5'-end-flanking region and 5,000 bp of the 3'-end-flanking region. The gene was shown to contain 39 exons. There were eight exons of 27 bp, five of 36 bp, four of 54 bp, three of 45 bp, and two of 42 bp. The rest of the exons coding for translated sequences had sizes varying between 24 and 153 bp. The genomic clones did not contain exons 3 and 4 whose sizes could, however, be estimated from cDNA clones. S1 nuclease mapping and primer extension analyzes indicated five closely spaced initiation sites of transcription. Sequencing of the 5'-end-flanking region did not reveal a typical TATA box but a four times repeated TATTTAT sequence that may serve as true TATA boxes. Two CCAAT boxes were found starting at positions-13 and -194, and furthermore, the promoter region contains two GC boxes. Previous studies on alpha 1 (XIII) collagen cDNA and genomic clones showed that the primary transcript undergoes complex alternative splicing generating at least four different forms of mRNAs. The present work demonstrated that sequences of seven exons are alternatively used. These exons contain sequences coding for pure collagenous regions, pure noncollagenous regions, and an exon coding for a junction of a collagenous and noncollagenous domain.


Is There an Association Between Circulating Kisspeptin Levels and Ovarian Reserve in Women of Reproductive Age?
To investigate the possible association of kisspeptin levels with the ovarian reserves of women of reproductive age. Eighty women aged 19-40 participated after signing an informed consent. Of these, 74 were finally included as in 6 women the blood samples were considered inappropriate due to hemolysis. They were divided into three main groups according to their ovarian reserve patterns: women with adequate ovarian reserves (Group A - AOR) (n=30), women with increased ovarian reserves (Group B - PCOS) (n=31), and women with diminished ovarian reserves (Group C - DOR) (n=13). Women with diminished ovarian reserves had statistically significantly increased age and FSH compared to the other two groups. No statistically significant difference was found between the three groups for estradiol and thyroid stimulating hormone. Moreover, body mass index, luteinizing hormone, total testosterone, 17-hydroxyprogesterone, dehydroepiandrosterone, anti-Mullerian hormone (AMH), and antral follicle count (AFC) were increased in group B compared to the other two groups. AMH and AFC were decreased in women with diminished ovarian reserves compared to the other two groups, as expected. The comparison of kisspeptin levels between the three groups showed that kisspeptin levels were increased in women with diminished ovarian reserves, compared to the other two groups, but without a statistically significant difference. However, kisspeptin levels in group C were statistically significantly higher than those in group A. There are no strong indications that kisspeptin levels are associated with the ovarian reserve in women of reproductive age.


Diversification of the plant-specific hybrid glycine-rich protein (HyGRP) genes in cereals.
Plant-specific hybrid proline- or glycine-rich proteins (HyP/GRPs) are involved in diverse gene functions including plant development and responses to biotic and abiotic stresses. The quantitative trait locus, qLTG3-1, enhances seed germination in rice under low-temperature conditions and encodes a member with a glycine-rich motif of the HyP/GRP family. The function of this gene may be related to the weakening of tissue covering the embryo during seed germination. In the present study, the diversification of the HyP/GRP gene family was elucidated in rice based on phylogenetic relationships and gene expression levels. At least 21 members of the HyP/GRP family have been identified in the rice genome and clustered in five regions on four chromosomes by tandem and chromosomal duplications. Of these, OsHyPRP05 (qLTG3-1) and its paralogous gene, OsHyPRP21, had a glycine-rich motif. Furthermore, orthologous genes with a glycine-rich motif and the HyP/GRP gene family were detected in four genome-sequenced monocots: 12 in barley, 10 in Brachypodium, 20 in maize, and 28 in sorghum, using a BLAST search of qLTG3-1 as the query. All members of the HyP/GRP family in these five species were classified into seven main groups, which were clustered together in these species. These results suggested that the HyP/GRP gene family was formed in the ancestral genome before the divergence of these species. The collinearity of chromosomal regions around qLTG3-1 and its orthologous genes were conserved among rice, Brachypodium, sorghum, and maize, indicating that qLTG3-1 and orthologous genes conserve gene function during seed germination.


[Stability examination of the inclusion compounds of 8 Chinese medicinal volatile oils with beta-cyclodextrin].
The content, hence the stability of volatile oils and their main components in the inclusion compounds of 8 Chinese medicinal volatile oils with beta-cyclodextrin have been examined with different storage times. The 8 Chinese medicines are Vitex negundo var. heterophylla, Notopterygium incisum, Alpinia officinarum, Atractylodes lancea, Pogostenon cablin, Asarum heterotropoides var. mandshuricum, Schizonepeta tenuifolia and Amomum villosum.


Analytic estimation of transition between instantaneous eigenstates of quantum two-level system.
Transition amplitudes between instantaneous eigenstates of a quantum two-level system are evaluated analytically on the basis of a new parametrization of its evolution operator, which has recently been proposed to construct exact solutions. In particular, the condition under which the transitions are suppressed is examined analytically. It is shown that the analytic expression of the transition amplitude enables us, not only to confirm the adiabatic theorem, but also to derive the necessary and sufficient condition for quantum two-level system to remain in one of the instantaneous eigenstates.


2,2':6',2''-terpyridine substituted in the 4'-position by the solubilizing and sterically demanding tert-butyl group: a surprisingly new ligand.
We describe the synthesis and characterization of 4'-tert-butyl-2,2':6',2''-terpyridine (4'-(t)Butpy, 1), a convergent tpy ligand that exhibits both a sterically demanding and solubilizing 4'-substituent. In the solid state, molecules of 1 pack with alternating tpy and tert-butyl domains, and the bulky alkyl substituents prevent the molecules from engaging in the face-to-face π-interactions which are typical of simple tpy ligands. Instead, the predominant packing forces involve CH···N hydrogen bonds and weak CH···π contacts. The syntheses of the homoleptic complexes [M(1)(2)][PF(6)](2) (M = Fe, Co, Zn and Ru) and the heteroleptic [Ru(tpy)(1)][PF(6)](2) are described. The complexes have been fully characterized in solution, including the (1)H NMR spectroscopic characterization of the paramagnetic [Co(1)(2)][PF(6)](2). Cyclic voltammetric data are consistent with the tert-butyl substituent being slightly electron releasing. The single crystal structures of [Zn(1)(2)][PF(6)](2) and [Ru(1)(2)][PF(6)](2) have been determined; the compounds are essentially isomorphous. The packing of the cations is such that the tert-butyl substituents are accommodated in pockets between the tpy domains of adjacent cations, and as a consequence, the {M(tpy)(2)}-embrace that is a ubiquitous feature of many related structures is not observed.


Speech understanding in quiet and noise in bilateral users of the MED-EL COMBI 40/40+ cochlear implant system.
The purpose of the study was to investigate speech understanding in quiet and noise in subjects bilaterally implanted with multi-channel cochlear implants. Nine adults bilaterally implanted with MED-EL implants were included in the study. The subjects were tested in three conditions: with both implants, with the right implant only, and with the left implant only. Speech tests included monosyllables in quiet and sentences in noise (10 dB signal to noise ratio). Speech was presented from the front, and noise was presented from either 90 degrees or 270 degrees azimuth. All subjects reported benefit from bilateral stimulation. Speech scores for all subjects were higher with bilateral than with unilateral stimulation. The average score across subjects for sentence understanding was 31.1 percentage points higher with both cochlear implants compared with the cochlear implant ipsilateral to the noise, and 10.7 percentage points higher with both cochlear implants compared with the cochlear implant contralateral to the noise. The average score for recognition of monosyllabic words was 18.7 percentage points higher with both cochlear implants than with one cochlear implant. All of these differences in average scores were significant at the 5% level. Bilateral cochlear implantation provides a significant benefit in speech understanding in both quiet and noise.


Civil Code (Amendment) Act, 1988 (No. 4 of 1988), 21 May 1988.
This Act amends the Civil Code of Saint Lucia to do the following, among other things: 1) allow marriage to be celebrated by giving notice; 2) legalize divorce; 3) allow all children to be legitimated by the subsequent marriage of their parents; 4) remove differences in the treatment of children born within marriage and those born outside of marriage; 5) lower to 18 the age of majority and the age under which consent must be obtained in order to marry. A number of these changes are consolidations of earlier amendments made to the Civil Code.


Galanin mediates tumor-induced immunosuppression in head and neck squamous cell carcinoma.
Galanin receptor 2 (GALR2) plays a significant role in the progression of head and neck squamous cell carcinomas (HNSCC). Since there is virtually no information on immunomodulation mediated by its ligand in the tumor microenvironment, we assessed the effects of galanin on peripheral blood mononuclear cells (PBMCs). After verification of GALR2 expression and it activity in PBMCs we evaluated the effect of galanin and conditioned media from HNSCC cell lines silenced for galanin or antibody-depleted, on proliferation, apoptosis, cytokine expression and activation/differentiation of immune cells. We found that galanin alone and as a component of the HNSCC secretome decreased HNSCC cell proliferation and expression of pro-inflammatory cytokines (IFNγ, IL-12, IL-17A, IL-1α, IL-6 and TNF-α), whilst increasing apoptosis and expression of pro-tumoral cytokines/growth factors (IL-10, IL-4, PDGF and GM-CSF). T cell activation (using CD69 as activation marker) and anti-tumoral phenotypes in CD4+ T cells (Th1 and Th17) were found to be suppressed. In vivo, tumor growth was found to be increased in the presence of galanin-stimulated PBMCs. Data from The Cancer Genome Atlas (TCGA) revealed that high expression of galanin was associated with a reduced overall survival of patients with HNSCC. Our data indicate that galanin secreted by HNSCC cells exhibits immune-suppressive and pro-tumoral effects.


Two-step cross-linking method for identification of NF-kappaB gene network by chromatin immunoprecipitation.
The chromatin immunoprecipitation (ChIP) assay has recently been exploited as a powerful and versatile technique for probing protein-DNA interactions within the chromatin environment. In this method, intact cells are fixed with a reversible DNA-protein cross-linking agent (formaldehyde), and associated DNA is enriched by immunoprecipitating a target DNA binding protein. The bound DNA in the immune complexes is then used to identify that specific DNA binding protein's endogenous genomic targets. Nuclear factor kappaB (NF-kappaB) is a highly inducible transcription factor that controls genetic networks important for pathogen- or cytokine-induced inflammation, immune response, and cellular survival. In our studies of the genetic network under control of the inducible NF-kappaB transcription factor, we found that the conventional ChIP technique using a single formaldehyde cross-linking step did not reproducibly cross-link it to DNA. As a result, we have developed a novel ChIP assay using a two-step cross-linking procedure, incorporating N-hydroxysuccinimide (NHS)-ester-mediated protein-protein cross-linking prior to conventional DNA-protein cross-linking. We demonstrate that this technique is highly efficient, cross-linking virtually all NF-kappaB/Rel A into covalent complexes, resulting in quantitative and robust identification of inducible NF-kappaB family binding to a variety of validated NF-kappaB-dependent genomic targets. To demonstrate the general utility of this two-step cross-linking procedure, we performed enhanced capture of cytokine-inducible signal transducer and activator of transcription-3 (STAT3) binding to one of its known target genes. Our method represents a significant improvement in the efficiency of ChIP analysis in the study of endogenous targets for rare transcription factors.


The bacterial microbiota in the ceca of Capercaillie (Tetrao urogallus) differs between wild and captive birds.
The diet of wild capercaillie differs strongly between seasons. Particularly during winter, when energy demands are high and the birds forage solely on coniferous needles, microbial fermentations in the ceca are considered to contribute significantly to the energy requirement and to the detoxification of the resinous diet. Here, we present the first cultivation-independent analysis of the bacterial community in the cecum of capercaillie, using the 16S rRNA gene as a molecular marker. Cloning and fingerprinting analyses of cecum feces show distinct differences between wild and captive birds. While certain lineages of Clostridiales, Synergistetes, and Actinobacteria are most prevalent in wild birds, they are strongly reduced in individuals raised in captivity. Most striking is the complete absence of Megasphaera and Synergistes species in captive capercaillie, which are characterized by a large abundance of Gammaproteobacteria closely related to members of the genus Anaerobiospirillum, bacteria that are commonly connected with intestinal dysfunction. The community profiles of cecum content from wild birds differed between summer and winter season, and the cecum wall may be an important site for bacterial colonization. Our results corroborate the hypothesis that the bacterial community in the ceca of tetraonid birds changes in response to their highly specialized seasonal diets. Moreover, we propose that the observed differences in community profiles between wild and captive capercaillie reflects a disturbance in the bacterial microbiota that compromises the performance of the cecum and may be responsible for the high mortality of captive birds released into nature.


A New Elliptical Model for Device-Free Localization.
Device-free localization (DFL) based on wireless sensor networks (WSNs) is expected to detect and locate a person without the need for any wireless devices. Radio tomographic imaging (RTI) has attracted wide attention from researchers as an emerging important technology in WSNs. However, there is much room for improvement in localization estimation accuracy. In this paper, we propose a geometry-based elliptical model and adopt the orthogonal matching pursuit (OMP) algorithm. The new elliptical model uses not only line-of-sight information, but also non-line-of-sight information, which divides one ellipse into several areas with different weights. Meanwhile the OMP, which can eliminate extra bright spots in image reconstruction, is used to derive an image estimator. The experimental results demonstrate that the proposed algorithm could improve the accuracy of positioning by up to 23.8% for one person and 33.3% for two persons over some state-of-the-art RTI methods.


Structure of barium perchlorate trihydrate.
Ba(ClO4)2.3H2O, Mr = 390.28, hexagonal, P63/m, a = 7.277 (2), c = 9.656 (1) A, V = 442.9 (1) A3, Z = 2, F(000) = 368, D chi = 2.93 g cm3, lambda(Mo K alpha) = 0.71069 A, T = 295 K, mu = 52.60 cm1, 456 unique reflections, R = 0.023. The barium atoms in this structure are coordinated by six water oxygens, O(3), at 2.919 (1) A and by six perchlorate oxygens, O(2), at 3.026 (2) A in a slightly distorted icosahedral arrangement, the average Ba-O separation thus being approximately 2.97 A. The perchlorate ion just fails to have regular tetrahedral geometry within the experimental error, and has an average Cl-O bond length of 1.433 (6) A. Consistent with hydrogen bonding delineated on the basis of least-squares-refined hydrogen-atom positions, each axial [O(1)] perchlorate oxygen is hydrogen bonded to three water molecules and each trigonal [O(2)] perchlorate oxygen is hydrogen bonded to two water molecules; thus each perchlorate ion is hydrogen bonded to nine water molecules. Conversely, each water molecule [O(3)] oxygen is hydrogen bonded to six perchlorate ions to two through axial [O(1)] oxygens and to four through trigonal [O(2)] oxygens.


Real Time Polymerase Chain Reaction for Hepatitis B Screening in Donor Corneas in the Central Eye Bank of Iran.
The aim of this study was to report the results of the use of real-time polymerase chain reaction (PCR) for the diagnosis of hepatitis B virus (HBV) infection in cornea donors at the Central Eye Bank of Iran. Between 2014 and 2016, all cornea donors that had negative screening serologic results for hepatitis B (HB) surface antigen, HB surface antibody (Ab), hepatitis C virus Ab, human immune deficiency virus Ab, human T-cell leukemia virus Ab, and syphilis, and positive serology for HB core Ab were subjected to real-time PCR with a detection limit of 400 IU/mL to identify HBV DNA. Positive results for HBV DNA were considered occult HBV infections in these donors. Over the 3-year period, 122 out of 10448 cornea donors had negative screening serologic tests outside of HB core Ab. Of which, 90 cases were subjected to real-time PCR. Occult HBV was detected in 11 cases (12.2%), resulting in the rejection of the corresponding corneas. The remaining 79 cases (87.8%) had negative results for HBV DNA and the corresponding corneas were used for transplantation. Implementation of PCR for the detection of occult HBV in cornea donors is necessary to not only increase the security level of cornea donation but also minimize the rejection rate of donors that have isolated HB core Ab reactivity.


Potential Effect of COVID-19 on Maternal and Infant Outcome: Lesson From SARS.
The coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is highly infectious and its ongoing outbreak has been declared a global pandemic by the WHO. Pregnant women are susceptible to respiratory pathogens and the development of severe pneumonia, suggesting the urgent need to assess the potential maternal and infant outcome of pregnancy with COVID-19. The intrauterine vertical transmission potential of SARS-CoV-2 also remains controversial. Herein, we discuss the potential effect of COVID-19 on maternal and infant outcomes based on current studies, including those published in Chinese, in a total of 80 mothers with COVID-19 and 80 infants. We also comprehensively explored the mother-to-child transmission routes of SARS-CoV-2, in particular the route of intrauterine vertical transmission. Given SARS-CoV-2 is a sister to SARS-CoV, of the SARS-related coronavirus species, we made a comprehensive comparison between them to learn from experiences with SARS. Although there is no evidence supporting the intrauterine vertical transmission of SARS-CoV-2, our comprehensive analysis suggests that the adverse maternal and infant outcomes caused by COVID-19 cannot be underestimated. Further, we speculated that the inconsistency between nucleic acids and serological characteristics IgM to SARS-CoV-2 of infants' specimens may be caused by the disruption of the amniotic barrier by the inflammatory factors induced by SARS-CoV-2 infection. Our review is beneficial to understand the effect of SARS-CoV-2 on maternal and infant outcomes.


Computational model demonstrates that Ndc80-associated proteins strengthen kinetochore-microtubule attachments in metaphase.
Chromosome segregation is mediated by spindle microtubules that attach to the kinetochore via dynamic protein complexes, such as Ndc80, Ska, Cdt1 and ch-TOG during mitotic metaphase. While experimental studies have previously shown that these proteins and protein complexes are all essential for maintaining a stable kinetochore-microtubule (kMT) interface, their exact roles in the mitotic metaphase remains elusive. In this study, we employed experimental and computational methods in order to characterize how these proteins can strengthen kMT attachments in both nonload-bearing and load-bearing conditions, typical of prometaphase and metaphase, respectively. Immunofluorescence staining of HeLa cells showed that the levels of Ska and Cdt1 significantly increased from prometaphase to metaphase, while levels of the Ndc80 complex remained unchanged. Our new computational model showed that by incorporating binding and unbinding of each protein complex coupled with a biased diffusion mechanism, the displacement of a possible complex formed by Ndc80-Ska-Cdt1 is significantly higher than that of Ndc80 alone or Ndc80-Ska. In addition, when we incorporate Ndc80/ch-TOG in the model, rupture force and time of attachment of the kMT interface increases. These results support the hypothesis that Ndc80-associated proteins strengthen kMT attachments, and that the interplay between kMT protein complexes in metaphase ensures stable attachments.


Microscale Self-Assembly of Upconversion Nanoparticles Driven by Block Copolymer.
Lanthanide-based upconversion nanoparticles can convert low-energy excitation to high-energy emission. The self-assembled upconversion nanoparticles with unique structures have considerable promise in sensors and optical devices due to intriguing properties. However, the assembly of isotropic nanocrystals into anisotropic structures is a fundamental challenge caused by the difficulty in controlling interparticle interactions. Herein, we report a novel approach for the preparation of the chain-like assemblies of upconversion nanoparticles at different scales from nano-scale to micro-scale. The dimension of chain-like assembly can be fine-tuned using various incubation times. Our study observed Y-junction aggregate morphology due to the flexible nature of amphiphilic block copolymer. Furthermore, the prepared nanoparticle assemblies of upconversion nanoparticles with lengths up to several micrometers can serve as novel luminescent nanostructure and offer great opportunities in the fields of optical applications.


Genetic predisposition to adverse consequences of anti-cholinesterases in 'atypical' BCHE carriers.
Normal butyrylcholinesterase (BuChE), but not several of its common genetic variants, serves as a scavenger for certain anti-cholinesterases (anti-ChEs). Consideration of this phenomenon becomes urgent in view of the large-scale prophylactic use of the anti-ChE, pyridostigmine, during the 1991 Persian Gulf War, in anticipation of nerve gas attack and of the anti-ChE, tacrine, for improving residual cholinergic neurotransmission in Alzheimer's disease patients. Adverse symptoms were reported for subjects in both groups, but have not been attributed to specific causes. Here, we report on an Israeli soldier, homozygous for 'atypical' BuChE, who suffered severe symptoms following pyridostigmine prophylaxis during the Persian Gulf War. His serum BuChE and recombinant 'atypical' BuChE were far less sensitive than normal BuChE to inhibition by pyridostigmine and several other carbamate anti-ChEs. Moreover, atypical BuChE demonstrated 1/200th the affinity for tacrine of normal BuChE or the related enzyme acetylcholinesterase (AChE). Genetic differences among BuChE variants may thus explain at least some of the adverse responses to anti-ChE therapies.


Use of Advanced Artificial Intelligence in Forensic Medicine, Forensic Anthropology and Clinical Anatomy.
Three-dimensional convolutional neural networks (3D CNN) of artificial intelligence (AI) are potent in image processing and recognition using deep learning to perform generative and descriptive tasks. Compared to its predecessor, the advantage of CNN is that it automatically detects the important features without any human supervision. 3D CNN is used to extract features in three dimensions where input is a 3D volume or a sequence of 2D pictures, e.g., slices in a cone-beam computer tomography scan (CBCT). The main aim was to bridge interdisciplinary cooperation between forensic medical experts and deep learning engineers, emphasizing activating clinical forensic experts in the field with possibly basic knowledge of advanced artificial intelligence techniques with interest in its implementation in their efforts to advance forensic research further. This paper introduces a novel workflow of 3D CNN analysis of full-head CBCT scans. Authors explore the current and design customized 3D CNN application methods for particular forensic research in five perspectives: (1) sex determination, (2) biological age estimation, (3) 3D cephalometric landmark annotation, (4) growth vectors prediction, (5) facial soft-tissue estimation from the skull and vice versa. In conclusion, 3D CNN application can be a watershed moment in forensic medicine, leading to unprecedented improvement of forensic analysis workflows based on 3D neural networks.


Underdetection and Undertreatment of Dyspnea in Critically Ill Patients.
Rationale: Dyspnea is a common and distressing physical symptom among patients in the ICU and may be underdetected and undertreated. Objectives: To determine the frequency of dyspnea relative to pain, the accuracy of nurses and personal caregiver dyspnea ratings relative to patient-reported dyspnea, and the relationship between nurse-detected dyspnea and treatment. Methods: This was an observational study of patients (n = 138) hospitalized in a medical ICU (MICU). Nurses and patients' personal caregivers at the bedside reported on their perception of patients' symptoms. Measurements and Main Results: Dyspnea was assessed by patients, caregivers, and nurses with a numerical rating scale. Across all three raters, the frequency of moderate to severe dyspnea was similar or greater than that of pain (P < 0.05 for caregiver and nurse ratings). Personal caregivers' ratings of dyspnea had substantial agreement with patient ratings (κ = 0.65, P < 0.001), but nurses' ratings were not significantly related to patient ratings (κ = 0.19, P = 0.39). Nurse detection of moderate to severe pain was significantly associated with opioid treatment (odds ratio, 2.70; 95% confidence interval, 1.10-6.60; P = 0.03); however, nurse detection of moderate to severe dyspnea was not significantly associated with any assessed treatment. Conclusions: Dyspnea was reported at least as frequently as pain among the sampled MICU patients. Personal caregivers had good agreement with patient reports of moderate to severe dyspnea. However, even when detected by nurses, dyspnea appeared to be undertreated. These findings suggest the need for improved detection and treatment of dyspnea in the MICU.


Transmembrane regions V and VI of the human luteinizing hormone receptor are required for constitutive activation by a mutation in the third intracellular loop.
Gonadotropin receptors are members of the seven-transmembrane (TM) receptor family. Several point mutations in TM V and VI and the intracellular loop 3 (i3) have been identified in the luteinizing hormone (LH) receptor gene, leading to constitutive activation of the receptor. Because gonadotropin receptors are highly conserved, we mutated the follicle-stimulating hormone (FSH) receptor at the corresponding amino acids. However, the FSH receptor mutants showed minimal increases in basal cAMP production. Taking advantage of this difference between the two receptors, we designed chimeric receptors with or without a point mutation in the i3 to identify the region in the LH receptor important for its constitutive activation. Introduction of the point mutation into chimeric receptors containing only TM V to VI from the LH receptor led to major increases in ligand-independent cAMP production. Furthermore, a chimeric receptor with only TM V and VI derived from the LH receptor can be rendered constitutively active by the mutation in the i3 from the FSH receptor. These results suggest that interactions between TM V and VI of the FSH receptor are essential for maintaining the receptor in the more constrained state, whereas interactions between these domains of the LH receptor are permissive for constitutively activating mutations in the i3.


Ethical issues for cancer screening.
In recent years medical ethics has become an undisputed part of medical studies. Many people believe that modern advances in medical technology - such as the development of dialysis machines, respirators, magnetic resonance imaging and genetic testing and types of cancer screenings - have created bioethical dilemmas that confront physicians in the 21st century. Debates over research and screening ethics have until recently revolved around two related questions: the voluntary, informed consent of subjects, and the appropriate relationship between risk and benefit to subjects. Every patient has a right to full and accurate information about his or her medical condition. This legal principle arose primarily through court decisions concerning informed consent, but over time physicians recognized that most patients prefer to learn the truth about their condition and use the information well. To screen is to search for disease in the absence of symptoms or, in other words, to attempt to find disease in someone not thought to have a disease. Examples of screening include routine mammography to detect breast cancer, routine pap smears to detect cervical cancer, and routine Prostate Specific Antigen (PSA) testing to detect prostate cancer. Ethical principles to be followed in cancer screening programmes are intended mainly to minimize unnecessary harm for the participating individuals. Numerous ethical questions can be raised about the practice of screening for disease. Here, we examine four leading cancer killers worldwide and we review the screening of protocols of these cancer types and their possible ethics.


Inspection Systems and Mechanical Removal Systems for Bone and Cartilage Particles in Ground Beef Patties.
Ground beef patties were processed from boneless beef which was subjected to two different inspection systems for bone and cartilage defects. One system permitted a higher ("high") level of defects (USDA Food Safety and Inspection Service, Meat and Poultry Inspection Requirements), while the other system (USDA PP-B-2120 Federal Purchasing Specification) permitted only a lower ("low") level of defects. Before formation into patties, the ground beef was subjected to three processing systems for defect removal (none, Weiler Bone Collector System, Speco Spiral Groove Plate System). A sensory approach (teeth and tongue) for detecting defects with cooked patty samples revealed more bone and cartilage defects in the "high" than the "low" formulation regardless of defect removal system. While the defect removal systems reduced the levels of detected defects in the "high" formulation, neither system produced patties with defect levels achieved through inspection procedures required to produce "low" levels of these defects. However, it appears that much of the bone remaining as a defect in boneless beef is reduced in size during grinding so as to be undetectable by sensory approaches.


Pyridine-functional diblock copolymer nanoparticles synthesized via RAFT-mediated polymerization-induced self-assembly: effect of solution pH.
Polymerization-induced self-assembly (PISA) via reversible addition-fragmentation chain transfer (RAFT) polymerization has become widely recognized as a versatile and efficient strategy to prepare complex block copolymer nanoparticles with controlled morphology, size, and surface functionality. In this article, we report the preparation of cationic sterically-stabilized poly(2-vinylpyridine)-poly(benzyl methacrylate) (P2VP-PBzMA) diblock copolymer nanoparticles via RAFT-mediated PISA under aqueous emulsion polymerization conditions. It is demonstrated that the solution pH during PISA has a dramatic effect on the resulting P2VP-PBzMA nanoparticles, as judged by dynamic light scattering (DLS), disc centrifuge photosedimentometry (DCP) and transmission electron microscopy (TEM). Varying the solution pH results in the P2VP stabilizer having different solubilities due to protonation/deprotonation of the pyridine groups. This allows P2VP-PBzMA nanoparticles with tunable diameters to be prepared by altering the DP of the stabilizer (P2VP) and/or core-forming block (PBzMA), or simply by changing the solution pH for a fixed copolymer composition. For example, P2VP-PBzMA nanoparticles with larger diameters can be obtained at higher solution pH as the protonation degree of the P2VP stabilizer has a large effect on both the aggregation of polymer chains during the PISA process, and the resulting behavior of the diblock copolymer nanoparticles. Changing the dispersion pH post-polymerization has a relatively limited effect on particle diameter. Furthermore, aqueous electrophoresis studies indicate that these P2VP-PBzMA nanoparticles had good colloidal stability and high cationic charge (>30 mV) below pH 5 and can be dispersed readily over a wide pH range.