Treatment of metastatic nonseminomatous germ cell tumors of the testis: significance of the international consensus prognostic classification as a prognostic factor-based staging system.
We reviewed treatment results in patients with metastatic nonseminomatous germ cell tumors of the testis and examined the significance of the International Consensus Prognostic Classification to make appropriate risk-based decisions concerning induction chemotherapy. We divided 37 patients treated with platinum-based combination chemotherapy into good, intermediate, and poor prognostic groups utilizing the International Consensus Prognostic Classification. The data was analyzed for both overall survival and progression-free survival among the 3 prognostic groups. Among the 37 patients, 10 died (8 of progressive disease, 1 of pneumonia during induction chemotherapy and 1 of cyclophosphamide-induced hemorrhagic cardiomyolitis during salvage chemotherapy). The survivors were followed for 6 to 1 84 months from the beginning of induction chemotherapy (median, 80 months). Five of the 37 patients (14%) were classified as having a good prognosis, 1 8 (48%) as intermediate, and 14 (38%) as having a poor prognosis. The patients in the poor prognostic group had a 5-year overall survival of only 40%, while those in the good and intermediate groups had 5-year overall survivals of 100% and 94%, respectively. When we applied the International Consensus Prognostic Classification to patients with advanced disease classified by the Indiana University Staging System, these patients could be clearly divided into good-risk and poor-risk groups. The International Consensus Prognostic Classification is easily applicable and accurate for risk assessment in patients with metastatic nonseminomatous germ cell tumors of the testis. This classification will now be widely used in general oncology practices and for clinical trials in these patients.


Enrichment-induced exercise to quantify the effect of different housing conditions: a tool to standardize enriched environment protocols.
Enriched environments (EE) have been used for a long time to promote recovery in many neurological disorders, however, a growing body of inconsistent results strongly calls for a rigorous standardization of experimental EE paradigms. Although some core principles are well accepted as standards, a method to quantitatively assess the complexity of EE in various experimental designs is still lacking. In this study, we tracked and recorded the physical exercise of rats in four housing conditions, namely isolated condition, social condition, novel condition and EE. Then, we analyzed whether and to what extent, enrichment-induced exercise reflected the degree of enrichment. We next examined rat exercise in a conventional environment condition and under different light intensities, to explore whether environment-related exercise could be considered a parameter to quantify the degree of enrichment. The results obtained showed that (1) both inanimate and social stimulations enhanced the exercise level and (2) EE combined the effects of the two stimulations. Furthermore, exercise durability which correlated positively with degree of enrichment, was an objective measure of different housing conditions. Exercise-related parameters also sensitively reflected the impacts of light intensity even in the same enrichment arrangements. Our results indicate that there is a direct and measurable correlation between degree of environmental enrichment and enrichment-induced exercise, and therefore enrichment-induced exercise could be used as a helpful tool to evaluate the degree of housing conditions and to standardize the EE protocols.


Clinical examination of the crystalline lens by retrocorneal specular microscopy.
Normal crystalline lenses and a variety of primary and secondary cataracts were photographed with a contact clinical specular microscope having a new W10X long working distance objective. In the normal lens, the anterior and posterior cell pattern and the lens fiber pattern could be seen. In patients with developing cataracts, a variety of abnormal structures was observed; in some cases the detailed morphologic structure of the light scattering objects could be appreciated. In pseudophakic patients, precipitates, leukocytes, and debris were observed on the surface of IOL implants. The methods described enable cellular and morphologic changes occurring with time in the lens and capsule to be studied. The results obtained show the potential usefulness of specular microscopy for evaluation and documentation of retrocorneal pathologic structures at high magnification.


Recent advances in the biomolecules mediated synthesis of nanoclusters for food safety analysis.
The development of nanoclusters based on incorporating biomolecules like proteins, lipids, enzymes, DNA, surfactants, and chemical stabilizers creates a stable and high fluorescence bio-sensors promising future due to their high sensitivity, high level of detection and better selectivity. This review addresses a comprehensive and systematic overview of the recent development in synthesizing metal nanocluster by various strategized synthesis techniques. Significantly, the application of nanometal clusters for the detection of various food contaminants such as microorganisms, antibodies, drugs, pesticides, metal contaminants, amino acids, and other food flavors have been discussed briefly concerning the detection techniques, sensitivity, selectivity, and lower limit of detection. The review further gives a brief account on the future prospects in the synthesis of novel metal nanocluster-based biosensors, and their advantages, shortcomings, and potential perspectives toward their application in the field of food safety analysis.


Analysis of N-glycosylation of phospholipase A2 from venom of individual bees by microbore high-performance liquid chromatography-electrospray mass spectrometry using an ion trap mass spectrometer.
The N-linked oligosaccharides were released from the phospholipase A2 (PLA) with glycopeptidases and reductively aminated with the chromophore, p-aminobenzoic acid ethyl ester (ABEE). The ABEE-labeled oligosaccharides were separated by microbore high-performance liquid chromatography (micro-HPLC) using a reversed-phase column and analyzed by electrospray mass spectrometry. Differentiation between alpha-1,3 and alpha-1,6 core-fucosylated glycans was achieved by comparison the glycans released by glycopeptidases peptide-N-glycanase A (PNase A) and peptide-N-glycanase F (PNase F). All N-linked oligosaccharides except 3B and 3C could be identified in this approach. The analysis of PLA oligosaccharides from the venom of individual bees indicated that glycosylation patterns between the younger and the older bees were similar.


Real-time seizure prediction using RLS filtering and interpolated histogram feature based on hybrid optimization algorithm of Bayesian classifier and Hunting search.
Epileptic seizure prediction using EEG signal analysis is an important application for drug therapy and pediatric patient monitoring. Time series estimation to obtain the future samples of EEG signal has vital role for detecting seizure attack. In this paper, a novel density-based real-time seizure prediction algorithm based on a trained offline seizure detection algorithm is proposed. In the offline seizure detection procedure, after signal preprocessing, histogram-based statistical features are extracted from signal probability distribution. By defining a deterministic polynomial model on the normalized histogram, a novel syntactic feature that is named Interpolated Histogram Feature (IHF) is proposed. Moreover, with this feature, Seizure Distribution Model (SDM) as a descriptor of the seizure and non-seizure signals is presented. By using a novel hybrid optimization algorithm based on Bayesian classifier and Hunting Search (HuS) algorithm, the optimal features are selected. To detect the seizure attacks in the online mode, a Multi-Layer Perceptron (MLP) classifier is trained with the optimal features in the offline procedure. For online prediction, the enhanced Recursive Least Square (RLS) filter is applied to estimate sample-by-sample of the EEG signal. Also, a density-based signal tracking scenario is introduced to update and tune the parameters of RLS filtering algorithm. Our prediction algorithm is evaluated on 104 hours of EEG signals recorded from 23 pediatric patients. Our online signal prediction algorithm provides the accuracy rate of 86.56% and precision rate of 86.53% simultaneously using the trained MLP classifier from the offline mode. The recall rate of seizure prediction is 97.27% and the false prediction rate of 0.00215 per hour is achieved as well. Ultimately, the future samples of EEG signal are estimated, and the time of seizure signal prediction is also converged to 6.64 seconds. In our proposed real-time algorithm, by implementing a density-based signal tracking scenario, the future samples of signal with suitable time is predicted and the seizure is detected based on the optimal features from the IHF and histogram-based statistical features with acceptable performance.


Metabolic engineering of Corynebacterium glutamicum for anthocyanin production.
Anthocyanins such as cyanidin 3-O-glucoside (C3G) have wide applications in industry as food colorants. Their current production heavily relies on extraction from plant tissues. Development of a sustainable method to produce anthocyanins is of considerable interest for industrial use. Previously, E. coli-based microbial production of anthocyanins has been investigated extensively. However, safety concerns on E. coli call for the adoption of a safe production host. In the present study, a GRAS bacterium, Corynebacterium glutamicum, was introduced as the host strain to synthesize C3G. We adopted stepwise metabolic engineering strategies to improve the production titer of C3G. Anthocyanidin synthase (ANS) from Petunia hybrida and 3-O-glucosyltransferase (3GT) from Arabidopsis thaliana were coexpressed in C. glutamicum ATCC 13032 to drive the conversion from catechin to C3G. Optimized expression of ANS and 3GT improved the C3G titer by 1- to 15-fold. Further process optimization and improvement of UDP-glucose availability led to ~ 40 mg/L C3G production, representing a > 100-fold titer increase compared to production in the un-engineered, un-optimized starting strain. For the first time, we successfully achieved the production of the specialty anthocyanin C3G from the comparatively inexpensive flavonoid precursor catechin in C. glutamicum. This study opens up more possibility of C. glutamicum as a host microbe for the biosynthesis of useful and value-added natural compounds.


Ezetimibe potently reduces vascular inflammation and arteriosclerosis in eNOS-deficient ApoE ko mice.
Hypercholesterolemia is associated with decreased vascular nitric oxide bioavailability and deletion of endothelial nitric oxide synthase (eNOS) markedly accelerates atherosclerosis development in apolipoprotein E knockout (apoE ko) mice. The current study tests whether atheroprotection provided by a lipid lowering therapy with Ezetimibe depends on eNOS. ApoE ko and apoE/eNOS double ko (dko) mice received a high fat diet with or without 0.05% Ezetimibe. Ezetimibe significantly reduced plasma cholesterol concentrations and atherogenic lipoproteins in both genotypes to a similar extent. Moreover, the drug reduced vascular inflammation, as it significantly reduced vascular cell adhesion molecule-1 (VCAM-1) expression and vascular CD14 expression, a marker for mononuclear cell infiltration, in both genotypes. Neither NOS protein expression nor vascular reactivity of aortic rings was changed in apoE ko mice following Ezetimibe treatment. Significant lesion reduction was seen in Ezetimibe-treated male and female apoE ko and apoE/eNOS dko animals (p<or=0.05). Interestingly, the drug-mediated additional atheroprotection in male apoE ko, compared to male eNOS dko mice, suggesting that lipid lowering does provide additional eNOS-dependent atheroprotection in this experimental group. Lipid lowering with Ezetimibe potently reduces atherosclerosis and vascular inflammation independent of eNOS. Moreover, Ezetimibe did not exert any effects on eNOS protein expression or enzyme activity. However, additional atheroprotection by Ezetimibe was observed in eNOS competent apoE ko mice, suggesting that some of the drug's anti-atherosclerotic effects are mediated by the eNOS pathway.


Effect of short-chain alcohols on the physicochemical properties of d-phenothrin emulsions and their insecticidal activity against Aedes aegypti.
Oil-in-water (o/w) emulsion is utilized as an insecticide delivery system for mosquito control. However, evaporation inhibition adjuvant is needed to prevent fog drift, inhibit release of insecticidal actives and prolong suspension time. In the current study, we evaluated the effect of different short-chain alcohols, namely, propylene glycol, 1,3-propanediol, glycerol and crude glycerol, as adjuvants on the physicochemical properties of d-phenothrin o/w emulsion system. The bioactivity of optimized formulations containing 20 wt% glycerol (D1), 20 wt% propylene glycol (D2) and without added alcohol (negative control) were tested against larvae, pupae and adult Aedes aegypti (Ae. aegypti). It was found that propylene glycol produced smaller droplets at lower concentrations but poor long-term stability at higher concentrations, whereas glycerol had an appreciable effect on initial droplet size and stability with increasing concentration. According to the dose-response bioassays and room size chamber testing, the highest larvicidal, pupicidal and adulticidal activities were observed with D2, followed by D1 and negative control. Overall, the above study demonstrated improved emulsion stabilities and potency against Ae. aegypti larvae, pupae and adults using glycerol as adjuvant for effective mosquito control.


Src-like adaptor protein down-regulates T cell receptor (TCR)-CD3 expression by targeting TCRzeta for degradation.
Src-like adaptor protein (SLAP) down-regulates expression of the T cell receptor (TCR)-CD3 complex during a specific stage of thymocyte development when the TCR repertoire is selected. Consequently, SLAP-/- thymocytes display alterations in thymocyte development. Here, we have studied the mechanism of SLAP function. We demonstrate that SLAP-deficient thymocytes have increased TCRzeta chain expression as a result of a defect in TCRzeta degradation. Failure to degrade TCRzeta leads to an increased pool of fully assembled TCR-CD3 complexes that are capable of recycling back to the cell surface. We also provide evidence that SLAP functions in a pathway that requires the phosphorylated TCRzeta chain and the Src family kinase Lck, but not ZAP-70 (zeta-associated protein of 70 kD). These studies reveal a unique mechanism by which SLAP contributes to the regulation of TCR expression during a distinct stage of thymocyte development.


Detection of mitochondrial dysfunction by EPR technique in mouse model of dilated cardiomyopathy.
Tgalphaq44 mice with targeted overexpression of activated Galphaq protein in cardiomyocytes mimic many of the phenotypic characteristics of dilated cardiomyopathy in humans. However, it is not known whether the phenotype of Tgalphaq44 mice would also involve dysfunction of cardiac mitochondria. The aim of the present work was to examine changes in EPR signals of semiquinones and iron in Fe-S clusters, as compared to classical biochemical indices of mitochondrial function in hearts from Tgalphaq44 mice in relation to the progression of heart failure. Tgalphaq44 mice at the age of 14 months displayed pulmonary congestion, increased heart/body ratio and impairment of cardiac function as measured in vivo by MRI. However, in hearts from Tgalphaq44 mice already at the age of 10 months EPR signals of semiquinones, as well as cyt c oxidase activity were decreased, suggesting alterations in mitochondrial electron flow. Furthermore, in 14-months old Tgalphaq44 mice loss of iron in Fe-S clusters, impaired citrate synthase activity, and altered mitochondrial ultrastructure were observed, supporting mitochondrial dysfunction in Tgalphaq44 mice. In conclusion, the assessment of semiquinones content and Fe(III) analysis by EPR represents a rational approach to detect dysfunction of cardiac mitochondria. Decreased contents of semiquinones detected by EPR and a parallel decrease in cyt c oxidase activity occurs before hemodynamic decompensation of heart failure in Tgalphaq44 mice suggesting that alterations in function of cardiac mitochondria contribute to the development of the overt heart failure in this model.


Migration behavior of metal complexes in capillary zone electrophoresis. Interpretation in terms of quantitative structure-mobility relationships.
Significant advances in metal ion analysis by capillary zone electrophoresis (CZE) have occurred as a consequence of using metal complexes with various organic and inorganic ligands. Metal-ligand complexes that contribute to metal speciation in solution have also gained the attention of researchers. An understanding of the molecular properties that control the separation and further insight into the migration mechanism call for a systematic analysis of relationships between migration parameters and charge and size characteristics of metal complexes. To perform such an investigation, a number multiparametric migration models derived from a generally valid equation for electrophoretic mobility as a function of charge density were developed. The models operating with tabular or readily calculated structural descriptors (in particular, metal atom electronegativity or effective charge) as well as with the formal charge and ligand number were evaluated using numerous sets of experimental migration data for inorganic and organic ligand complexes. Consistent--in a great many instances--approximation results confirm the separation mechanism for metal complexes in CZE as governed basically by differences in charge-to-size parameters, present a valuable and convincing selection of such parameters, possessing a definite physical meaning, and owing to the general validity of the multivariate regression approach, open new possibilities in its application to more complex (i.e., electrokinetic chromatographic) systems.


Glossopharyngeal Nerve Block with Long Acting Local Anaesthetic Agent (Bupivacaine) and It's Effect on Early Post-operative Period in Adult Tonsillectomy: A Prospective Study.
We evaluated the benefit of glossopharyngeal nerve block with long acting local anaesthetic like bupivacaine. It was a randomized prospective study. Sixty-four patients were selected and divided into two groups. Group A received bilateral nerve block and Group B received no block. Pain score using Visual Analog Scale (0-100 mm) was assessed at 30 min, 2, 6 and 12 h. In the immediate post operative period pain scores of Group A at rest and swallowing was significantly lower than Group B (p < 0.001 and p < 0.01). Glossopharyngeal nerve block is an important method of reducing post-tonsillectomy pain.


Severe MR With Prior Alfieri Stitch Treated With Transcatheter Edge-to-Edge Repair and a Vascular Plug.
We present a case of a high-risk surgical patient with prior surgical Alfieri stitch and recurrent severe mitral regurgitation. In cases with suitable anatomy, mitral valve transcatheter edge-to-edge repair and vascular plug closure of a small regurgitant orifice can be used with excellent results. (Level of Difficulty: Advanced.).


[Effect of lithium hydroxybutyrate on the serotonin content of the brain in intact rabbits and during hyperactivity of the central serotoninergic systems].
Chronic administration of lithium hydroxybutyrate (10 mg/kg) to rats reduces the content of serotonin in the caudate nucleus, suppresses the growth of its level in all the subcortical brain structures. After 5-hydroxytryptophan administration it perverses the serotonin-positive action of dorsal raphe nucleus electric stimulation.


Evaluation of the Implementation of the Response Assessment in Neuro-Oncology Criteria in the HERBY Trial of Pediatric Patients with Newly Diagnosed High-Grade Gliomas.
HERBY was a Phase II multicenter trial setup to establish the efficacy and safety of adding bevacizumab to radiation therapy and temozolomide in pediatric patients with newly diagnosed non-brain stem high-grade gliomas. This study evaluates the implementation of the radiologic aspects of HERBY. We analyzed multimodal imaging compliance rates and scan quality for participating sites, adjudication rates and reading times for the central review process, the influence of different Response Assessment in Neuro-Oncology criteria in the final response, the incidence of pseudoprogression, and the benefit of incorporating multimodal imaging into the decision process. Multimodal imaging compliance rates were the following: diffusion, 82%; perfusion, 60%; and spectroscopy, 48%. Neuroradiologists' responses differed for 50% of scans, requiring adjudication, with a total average reading time per patient of approximately 3 hours. Pseudoprogression occurred in 10/116 (9%) cases, 8 in the radiation therapy/temozolomide arm and 2 in the bevacizumab arm (P < .01). Increased target enhancing lesion diameter was a reason for progression in 8/86 cases (9.3%) but never the only radiologic or clinical reason. Event-free survival was predicted earlier in 5/86 (5.8%) patients by multimodal imaging (diffusion, n = 4; perfusion, n = 1). The addition of multimodal imaging to the response criteria modified the assessment in a small number of cases, determining progression earlier than structural imaging alone. Increased target lesion diameter, accounting for a large proportion of reading time, was never the only reason to designate disease progression.


Use of a Paid Digital Marketing Campaign to Promote a Mobile Health App to Encourage Parent-Engaged Developmental Monitoring: Implementation Study.
The internet has become an increasingly popular medium for parents to obtain health information. More studies investigating the impact of paid digital marketing campaigns for parents on promoting children's healthy development are needed. This study aims to explore the outcomes of a paid digital marketing campaign, which occurred from 2018 to 2020, to promote messages about parent-engaged developmental monitoring and ultimately direct parents to the Centers for Disease Control and Prevention's (CDC's) Milestone Tracker app, a mobile health (mHealth) app developed by the CDC. The paid digital marketing campaign occurred in 3 phases from 2018 to 2020. In each phase, 24 to 36 marketing messages, in English and Spanish, were created and disseminated using Google's Universal App Campaigns and Facebook Ads Manager. Outcomes were measured using impressions, clicks, and install data. Return on investment was measured using click-through rate (CTR), cost per click, and cost per install metrics. The Google-driven marketing messages garnered a total of 4,879,722 impressions (n=1,991,250, 40.81% for English and n=2,888,472, 59.19% for Spanish). The messages resulted in a total of 73,956 clicks (n=44,328, 59.94% for English and n=29,628, 40.06% for Spanish), with a total average CTR of 1.52% (2.22% for English and 1.03% for Spanish). From these clicks, there were 13,707 installs (n=9765, 71.24% for English and n=3942, 28.76% for Spanish) of the CDC's Milestone Tracker app on Google Play Store. The total average cost per install was US $0.93 across all phases. The phase 3 headline "Track your child's development" generated the highest CTR of 3.23% for both English and Spanish audiences. The Facebook-driven marketing messages garnered 2,434,320 impressions (n=1,612,934, 66.26% for English and n=821,386, 33.74% for Spanish). The messages resulted in 44,698 clicks (n=33,353, 74.62% for English and n=11,345, 25.38% for Spanish), with an average CTR of 1.84% (2.07% for English and 1.38% for Spanish). In all 3 phases, animated graphics generated the greatest number of clicks among both English and Spanish audiences on Facebook when compared with other types of images. These paid digital marketing campaigns can increase targeted message exposure about parent-engaged developmental monitoring and direct a parent audience to an mHealth app. Digital marketing platforms provide helpful metrics that can be used to assess the reach, engagement, and cost-effectiveness of this effort. The results from this study suggest that paid digital marketing can be an effective strategy and can inform future digital marketing activities to promote mHealth apps targeting parents of young children.


Flexible Fiber-Optic High-Speed Imaging of Vocal Fold Vibration: A Preliminary Report.
High-speed video (HSV) imaging of vocal fold vibration has been possible only through the rigid endoscope. This study reports that a fiberscope-based high-speed imaging system may allow HSV imaging of naturalistic voicing. Twenty-two subjects were recorded using a commercially available black and white high-speed camera (Photron Motion Tools, 256 × 120 pixel, 2000 frames per second, 8 second acquisition time). The camera gain is set to +6 db. The camera is coupled to a standard fiber-optic laryngoscope (Olympus ENF P-4) with a 300-W Xenon light. Image acquisition was done by asking the subject to perform repeated phonation at modal phonation. Video images were processed using commercial video editing and video noise reduction software (After effects, Magix, and Neat Video 4.1). After video processing, the video images were analyzed using digital kymography (DKG). The HSV black and white video acquired by the camera is gray and lacks contrast. By adjustment of image contrast, brightness, and gamma and using noise reduction software, the flexible laryngoscopy image can be converted to video image files suitable for DKG and waveform analysis. The increased noise still makes edge tracking for objective analysis difficult, but subjective analysis of DKG plot is possible. This is the first report of HSV acquisition in an unsedated patient using a fiberscope. Image enhancement and noise reduction can enhance the HSV to allow extraction of the digital kymogram. Further image enhancement may allow for objective analysis of the vibratory waveform.


Targeting cholesterol with β-cyclodextrin sensitizes cancer cells for apoptosis.
We found that targeting cholesterol with beta-cyclodextrin (bCD) and its derivatives disrupted signal transduction between PI3K and AKT, attenuating AKT pro-survival signals. In their absence, 2-deoxyglucose (2DG) caused anti-apoptotic protein Mcll to dissociate from pro-apoptotic Bak at mitochondria. Normally Bak is sequestered by its inhibitory associations with Mcll and Bcl-xL, and only when Bak is released from both, is it free to form oligomers through which cytochrome c can escape into the cytosol. Thus an addition of a bcl-2 antagonist dissociates Bak from Bcl-xL, triggering cytochrome c release and inducing apoptosis. 2DG-bCD can also sensitize type II cancer cells for TRAIL-mediated apoptosis.


In vitro and in vivo experimental trials to assess the modulatory influence of β-caryophyllene on NDV replication and immunopathogenesis.
A trial was conducted to evaluate the antiviral activity and immunomodulatory effect of B-Caryophyllene (BCP) using NDV as a viral model. First, an in ovo experiment was conducted to estimate the antiviral mechanism of BCP. Next, an in vivo experiment was designed to confirm its antiviral efficacy as well as its immunomodulatory and growth promoting ability. According to the in ovo experiment, BCP possesses antiviral influence up to 61.7% when treated before or during NDV infection. Oral supplementation of chickens with two doses of BCP (200 and 400 μg/bird) resulted in a significant increase in the NDV HI-Ab responses and a significant increase in interferon-α signaling cytokines. These obvious immunomodulatory effects improved the bird clinical protection against virulent NDV challenge. To conclude, we introduced a new compound for the poultry industry sector that has antiviral and immunostimulant properties when supplemented orally before or during NDV infection.


Unique Immune Cell Coactivators Specify Locus Control Region Function and Cell Stage.
Locus control region (LCR) functions define cellular identity and have critical roles in diseases such as cancer, although the hierarchy of structural components and associated factors that drive functionality are incompletely understood. Here we show that OCA-B, a B cell-specific coactivator essential for germinal center (GC) formation, forms a ternary complex with the lymphoid-enriched OCT2 and GC-specific MEF2B transcription factors and that this complex occupies and activates an LCR that regulates the BCL6 proto-oncogene and is uniquely required by normal and malignant GC B cells. Mechanistically, through OCA-B-MED1 interactions, this complex is required for Mediator association with the BCL6 promoter. Densely tiled CRISPRi screening indicates that only LCR segments heavily bound by this ternary complex are essential for its function. Our results demonstrate how an intimately linked complex of lineage- and stage-specific factors converges on specific and highly essential enhancer elements to drive the function of a cell-type-defining LCR.


The sequence of events in the differentiation of the epidermis in fetal rats with particular reference to membrane-coating granules.
The sequence of events in the establishment of a keratinised epidermis has been related to age and crown rump length in fetal rats. Differentiation of the epidermis occurs under cover of the periderm throughout gestation. Ten stages are defined between 12d intra-uterine life and birth. Membrane-coating granules (MCGs) appeared at 18d (Stage 4) after the appearance of tonofilaments but before the appearance of the first "fetal" keratohyaline granules (KHGs) at 19d (Stage 5). Measurement of the position of MCGs within the cells showed a less marked concentration near the superficial border of the cells than that found at later stages. As KHGs formed, exocytosis of MCGs occurred into the intercellular space immediately deep to the periderm. It is suggested that after 19d (Stage 5) the periderm serves to retain the contents of the MCGs in this space so providing the permeability barrier and that prior to that stage the periderm itself probably provides the permeability barrier of fetal skin.


[Morphology of the adult human hyoid bone].
Investigations on 504 human hyoid bones show that there exist three main types, whose occurrence is, to a certain degree, sex-related. The junction between the body and the larger horns of the hyoid often is not ossified even in advanced age. Any determination of age on the basis of the extent of the corpus-cornu maius-ossification is most unreliable.


Common feline problem behaviors: Aggression in multi-cat households
Being able to understand and intervene in cases of cat-cat aggression in multi-cat households is important for all veterinarians and behavior counsellors dealing with feline behavior cases. Feline conflicts are common, perhaps not helped by many owners' assumption that this is the norm. Interventions can be complex in multi-cat households with large numbers of cats as there are more interactions to evaluate and monitor, and care needs to be taken not to negatively affect the other cats. There may be limitations due to the environment or the people involved and so behavior guidelines should always be tailored to the individual situation. This article reviews the main forms of aggression in multi-cat households and the appropriate behavioral interventions, considering both the typical scenarios and emotions/motivations most likely involved. It details essential environmental management for multi-cat homes, as well as an approach to friendly cat introductions. This article is aimed at veterinarians dealing with feline behavior cases as well as behavior counsellors, as both professionals are likely to experience cases of aggression in multi-cat households. The author draws on the published literature where available and, where there is a lack of research, on hypotheses derived from her own clinical behavior experience.


Plasma-triglycerides and exercise: a delicate balance.
Alimentary lipemia was studied in 12 healthy young men with and without exercise. Three sets of experiments were performed. While continuous exercise of 90 min duration significantly reduced postprandial triglycerides by 26% (study I), this effect could not be observed when exercise was interrupted for 5 min after each 25 min (study II). Plasma free fatty acid concentrations, in the latter experiment, were significantly higher (by 311%) than during rest. When, in a third experiment continuous exercise was compared with intermittent physical activity, the latter condition significantly increased postprandial triglyceridemia, most probably due to precipitous rises of free fatty acids on each interruption of ergometry. It is concluded that in the third experiment the balance between triglyceride removal and triglyceride synthesis was shifted toward the latter. Whether exercise lowers, leaves unaltered, or raises plasma triglyceride levels may depend on subtle changes of experimental design.


Transcriptome profiles of metamorphosis in the ornamented pygmy frog Microhyla fissipes clarify the functions of thyroid hormone receptors in metamorphosis.
Anuran metamorphosis is an excellent system in which to study postembryonic development. Studies on Xenopus (Mesobatrachia) show that thyroid hormone receptors (TRs) regulate metamorphosis in a ligand-dependent manner by coordinating the action of hundreds of genes. However, whether this mechanism is conserved among amphibians is still unknown. To understand the molecular mechanism of this universal phenomenon, we report the transcriptional profiles of the three key developmental stages in Microhyla fissipes (Neobatrachia): premetamorphosis (PM), metamorphic climax (MC) and completion of metamorphosis (CM). In total, 2,293 differentially expressed genes were identified from comparisons of transcriptomes, and these genes showed stage-specific expression patterns. Unexpectedly, we found that TRα was highly expressed in Xenopus laevis and Bufo gargarizans at premetamorphosis but showed low expression in M. fissipes. In contrast, TRβ was highly expressed during metamorphosis in M. fissipes and X. laevis. This result may imply that TRβ is essential for initiating metamorphosis, at least in M. fissipes. Thus, our work not only identifies genes that are likely to be involved in Neobatrachia metamorphosis but also clarifies the roles of unliganded TRα in regulating tadpole growth and timing of metamorphosis, which may be conserved in anurans, and the role of liganded TRβ in launching metamorphosis.


The CD4/CD8 lineages: central decisions and peripheral modifications for T lymphocytes.
CD4(+) helper and CD8(+) cytotoxic T cells, two major subsets of αβTCR expressing lymphocytes, are differentiated from common precursor CD4(+)CD8(+) double-positive (DP) thymocytes. Bifurcation of the CD4(+)/CD8(+) lineages in the thymus is a multilayered process and is thought to culminate in a loss of developmental plasticity between these functional subsets. Advances in the last decade have deepened our understanding of the transcription control mechanisms governing CD4 versus CD8 lineage commitment. Reciprocal expression and antagonistic interplay between two transcription factors, ThPOK and Runx3, is crucial for driving thymocyte decisions between these two cell fates. Here, we first focus on the regulation of ThPOK expression and its role in directing helper T cell development. We then discuss a novel aspect of the ThPOK/Runx3 axis in modifying CD4(+) T cell function upon exposure to gut microenvironment.


Removing rician bias in diffusional kurtosis of the prostate using real-data reconstruction.
To compare prostate diffusional kurtosis imaging (DKI) metrics generated using phase-corrected real data with those generated using magnitude data with and without noise compensation (NC). Diffusion-weighted images were acquired at 3T in 16 prostate cancer patients, measuring 6 b-values (0-1500 s/mm2 ), each acquired with 6 signal averages along 3 diffusion directions, with noise-only images acquired to allow NC. In addition to conventional magnitude averaging, phase-corrected real data were averaged in an attempt to reduce rician noise-bias, with a range of phase-correction low-pass filter (LPF) sizes (8-128 pixels) tested. Each method was also tested using simulations. Pixelwise maps of apparent diffusion (D) and apparent kurtosis (K) were calculated for magnitude data with and without NC and phase-corrected real data. Average values were compared in tumor, normal transition zone (NTZ), and normal peripheral zone (NPZ). Simulations indicated LPF size can strongly affect K metrics, where 64-pixel LPFs produced accurate metrics. Relative to metrics estimated from magnitude data without NC, median NC K were lower (P < 0.0001) by 6/11/8% in tumor/NPZ/NTZ, 64-LPF real-data K were lower (P < 0.0001) by 4/10/7%, respectively. Compared with magnitude data with NC, phase-corrected real data can produce similar K, although the choice of phase-correction LPF should be chosen carefully.


Clinical characteristics and treatment outcomes of cytomegalovirus anterior uveitis and endotheliitis: A systematic review and meta-analysis.
Cytomegalovirus (CMV) anterior uveitis is the most common form of ocular manifestation of CMV in immunocompetent individuals. The difficulty in diagnosing CMV anterior uveitis may delay adequate treatment and affect outcomes. We sought to review systemically the overall clinical characteristics and compare treatment outcomes in CMV anterior uveitis and endotheliitis. A literature search was performed, and studies describing clinical characteristics, treatment regimens, and outcomes that included more than 5 treated eyes were included. In these 23 studies, acute CMV anterior uveitis commonly presented with high intraocular pressure (95.31%, 95% CI 90.45-98.60) and mild anterior chamber inflammation (cells >2+ = 3.18%, 95% CI 0.21-0.54). About two-thirds of CMV endotheliitis cases presented with high intraocular pressure and coin-shaped corneal lesions. Acute CMV anterior uveitis showed good clinical response to topical 0.15% ganciclovir (GCV) gel or oral valganciclovir (VGCV) (90%, 95% CI 74-100% and 95%, 95% CI 88-100%, respectively). For chronic CMV anterior uveitis, both topical GCV and oral VGCV yielded comparable results. Topical 0.5-2% GCV or a combination of topical and oral VGCV for CMV endotheliitis both resulted in good clinical response. Recurrence of inflammation was common after cessation of maintenance therapy. Overall, topical GCV resulted in an optimal outcome for CMV anterior uveitis. Escalated concentration and frequency of usage are needed for chronic CMV anterior uveitis and endotheliitis. Adequate induction and maintenance phases of anti-CMV treatment seem necessary to prevent recurrences.


Viral community-wide auxiliary metabolic genes differ by lifestyles, habitats, and hosts.
Viral-encoded auxiliary metabolic genes (AMGs) are important toolkits for modulating their hosts' metabolisms and the microbial-driven biogeochemical cycles. Although the functions of AMGs have been extensively reported in numerous environments, we still know little about the drivers that shape the viral community-wide AMG compositions in natural ecosystems. Exploring the drivers of viral community-wide AMG compositions is critical for a deeper understanding of the complex interplays among viruses, hosts, and the environments. Here, we investigated the impact of viral lifestyles (i.e., lytic and lysogenic), habitats (i.e., water, particle, and sediment), and prokaryotic hosts on viral AMG profiles by utilizing metagenomic and metatranscriptomic techniques. We found that viral lifestyles were the most important drivers, followed by habitats and host identities. Specifically, irrespective of what habitats viruses came from, lytic viruses exhibited greater AMG diversity and tended to encode AMGs for chaperone biosynthesis, signaling proteins, and lipid metabolism, which could boost progeny reproduction, whereas temperate viruses were apt to encode AMGs for host survivability. Moreover, the lytic and temperate viral communities tended to mediate the microbial-driven biogeochemical cycles, especially nitrogen metabolism, in different manners via AMGs. When focusing on each lifestyle, we further found clear dissimilarity in AMG compositions between water and sediment, as well the divergent AMGs encoded by viruses infecting different host orders. Overall, our study provides a first systematic characterization of the drivers of viral community-wide AMG compositions and further expands our knowledge of the distinct interactions of lytic and temperate viruses with their prokaryotic hosts from an AMG perspective, which is critical for understanding virus-host-environment interactions in natural conditions. Video Abstract.


Hypermethylation of Long Non-Coding RNA Genes Group in the Breast Cancer Development and Progression.
We analyzed changes in the level of methylation of CpG islands in four long non-coding RNA (lncRNA) genes MEG3, ZNF667-AS1, GAS5, and SEMA3B-AS1 as promising markers of breast cancer. Methylation analysis was performed by quantitative methylation-specific PCR on a set of 38 paired (tumor/normal) breast cancer samples. Significantly (p<0.001) increased methylation was shown for three of the four lncRNA genes: MEG3, ZNF667-AS1, and SEMA3B-AS1. We found significant correlations of the methylation level of all the studied lncRNA genes with the stage of cancer and with lymphogenic metastasis, and for MEG3 and ZNF667-AS1 also with the tumor size. Methylation of ZNF667-AS1, and SEMA3B-AS1 genes in breast cancer was detected for the first time. Based on these findings, new potential markers for the diagnosis and prognosis of breast cancer can be proposed.


Bimetallic CoCu nanoparticles anchored on COF/SWCNT for electrochemical detection of carbendazim.
Carbendazim (CBZ) is a widespread fungicide used in crop protection, but the CBZ residues in drinking water, fruits, and vegetables can also cause adverse impacts on public health due to direct exposure. In this paper, a ternary synergistic composite of bimetallic CoCu nanoparticles anchored on covalent organic framework/single-walled carbon nanotube (CoCu/COF/SWCNT) was prepared and further applied as an electrochemical sensing platform for detecting CBZ. The sensor showed a sensitive response performance toward CBZ oxidation, as a result of the enhanced charge transfer ability, large electrochemically active surface area, and high electro-catalytic activity from the rational integration of the ternary components in CoCu/COF/SWCNT. Under the optimal conditions, the proposed sensor exhibited a detection range of 0.001 to 10 μM and a limit detection of 0.65 nM for CBZ detection. In addition, the sensor displayed practical feasibility for the determination of CBZ in water and pear samples with a recovery of 96.1 % to 102.1 %.


Temporal changes in short-term associations between cardiorespiratory emergency department visits and PM2.5 in Los Angeles, 2005 to 2016.
Emissions control programs targeting certain air pollution sources may alter PM2.5 composition, as well as the risk of adverse health outcomes associated with PM2.5. We examined temporal changes in the risk of emergency department (ED) visits for cardiovascular diseases (CVDs) and asthma associated with short-term increases in ambient PM2.5 concentrations in Los Angeles, California. Poisson log-linear models with unconstrained distributed exposure lags were used to estimate the risk of CVD and asthma ED visits associated with short-term increases in daily PM2.5 concentrations, controlling for temporal and meteorological confounders. The models were run separately for three predefined time periods, which were selected based on the implementation of multiple emissions control programs (EARLY: 2005-2008; MIDDLE: 2009-2012; LATE: 2013-2016). Two-pollutant models with individual PM2.5 components and the remaining PM2.5 mass were also considered to assess the influence of changes in PM2.5 composition on changes in the risk of CVD and asthma ED visits associated with PM2.5 over time. The relative risk of CVD ED visits associated with a 10 μg/m3 increase in 4-day PM2.5 concentration (lag 0-3) was higher in the LATE period (rate ratio = 1.020, 95% confidence interval = [1.010, 1.030]) compared to the EARLY period (1.003, [0.996, 1.010]). In contrast, for asthma, relative risk estimates were largest in the EARLY period (1.018, [1.006, 1.029]), but smaller in the following periods. Similar temporal differences in relative risk estimates for CVD and asthma were observed among different age and season groups. No single component was identified as an obvious contributor to the changing risk estimates over time, and some components exhibited different temporal patterns in risk estimates from PM2.5 total mass, such as a decreased risk of CVD ED visits associated with sulfate over time. Temporal changes in the risk of CVD and asthma ED visits associated with short-term increases in ambient PM2.5 concentrations were observed. These changes could be related to changes in PM2.5 composition (e.g., an increasing fraction of organic carbon and a decreasing fraction of sulfate in PM2.5). Other factors such as improvements in healthcare and differential exposure misclassification might also contribute to the changes.


Accurate tumor purity determination is critical for the analysis of homologous recombination deficiency (HRD).
Homologous recombination deficiency (HRD) is a predictive marker for response to poly (ADP-ribose) polymerase inhibitors (PARPi) in ovarian carcinoma. HRD scores have entered routine diagnostics, but the influence of algorithms, parameters and confounders has not been analyzed comprehensively. A series of 100 poorly differentiated ovarian carcinoma samples was analyzed using whole exome sequencing (WES) and genotyping. Tumor purity was determined using conventional pathology, digital pathology, and two bioinformatic methods. HRD scores were calculated from copy number profiles determined by Sequenza and by Sclust either with or without fixed tumor purity. Tumor purity determination by digital pathology combined with a tumory purity informed variant of Sequenza served as reference method for HRD scoring. Seven tumors had deleterious mutations in BRCA1/2, 12 tumors had deleterious mutations in other homologous recombination repair (HRR) genes, 18 tumors had variants of unknown significance (VUS) in BRCA1/2 or other HRR genes, while the remaining 63 tumors had no relevant alterations. Using the reference method for HRD scoring, 68 tumors were HRD-positive. HRDsum determined by WES correlated strongly with HRDsum determined by single nucleotide polymorphism (SNP) arrays (R = 0.85). Conventional pathology systematically overestimated tumor purity by 8% compared to digital pathology. All investigated methods agreed on classifying the deleterious BRCA1/2-mutated tumors as HRD-positive, but discrepancies were observed for some of the remaining tumors. Discordant HRD classification of 11% of the tumors was observed comparing the tumor purity uninformed default of Sequenza and the reference method. In conclusion, tumor purity is a critical factor for the determination of HRD scores. Assistance by digital pathology helps to improve accuracy and imprecision of its estimation.


A double-blind, placebo-controlled, single-ascending dose study to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of HM15136, a novel long-acting glucagon analogue, in healthy subjects.
To evaluate the safety and tolerability, pharmacokinetics, and pharmacodynamics of HM15136, a novel long-acting glucagon analogue under development, in healthy males and females presenting with no childbearing potential. A randomized, double-blind, placebo-controlled, single-ascending dose study was conducted in 56 subjects who randomly received a single subcutaneous dose of HM15136 or its matching placebo at a ratio of 6:2 at 10, 20, 30, 50, 80, 100, and 120 μg/kg. All adverse events were mild and transient. Neither serious adverse events nor discontinuation as a result of adverse events occurred. The most frequent adverse drug reaction was nausea (5.3%, only in the 100- and 120-μg/kg groups). HM15136, particularly at doses of 50 μg/kg or higher, increased fasting blood glucose, with a maximum increase and area under the curve of 1.5 mmol/L at day 10 (P = .006) and 166.3 day·mmol/L (P = .022) at the dose of 80 μg/kg, while suppressing the secretion of endogenous glucagon, which continued until day 17. HM15136 also significantly reduced gluconeogenic and ketogenic amino acids. Compensatory changes in endogenous insulin and incretin hormones by HM15136 were not apparent. HM15136 was slowly but steadily absorbed and reached a peak concentration at 46-68 hours after a single subcutaneous injection. HM15136 was eliminated with a terminal phase half-life of 77.1-101.1 hours. A single subcutaneous dose of HM15136 at 10-120 μg/kg was safe and well tolerated. The long half-life of HM15136, coupled with an increase in blood glucose for ~2 weeks, may warrant a weekly dosing regimen.


Laparoscopic management of a true broad ligament leiomyoma in a patient with advanced endometriosis and a solitary kidney - A case report and literature review.
Leiomyomas are the most common benign tumours of the female genital tract, and almost always arise from the uterine myometrium. Although extrauterine leiomyomas are rare, they usually develop in sites such as the ovary, broad ligament, round ligament, cervix or abdominal wall. The broad ligament is the most common site of extrauterine leiomyoma involvement, and this unique clinical entity may prove to be a diagnostic or therapeutic challenge, particularly in patients with advanced endometriosis and distorted pelvic anatomy. Herein, we report the case of a large true broad ligament leiomyoma that was discovered during a total laparoscopic hysterectomy and bilateral salpingo-oophorectomy in a 47-year-old patient with stage IV endometriosis and a congenital left kidney and left ureter. This case highlights a rare occurrence of a true broad ligament leiomyoma, the challenges associated with preoperative diagnosis, and the laparoscopic approach to its management.


[Radiographic studies of cervical spine motion range in normal Chinese subjects with computer-assisted image analysis: full flexion and extension].
To study the degenerative regularity of normal Chinese cervical spine by observing the motion range of full cervical flexion and extension, and to provide evidences for further investigation of cervical segmental instability. 764 X-ray plain films of full cervical flexion and extension in 382 normal Chinese subjects (214 were male and 168 female) were investigated using self-designed cervical X-ray image analysis system. The subjects within the same sex group were divided into three groups according to the age: group A (20 years to 39 years), group B (40 years to 59 years) and group C (60 years to 76 years). During the full cervical flexion and extension motion, total intervertebral motion range (from C(2) - C(3) to C(6) - C(7)) and intervertebral motion range at each level were major parameters. The range of total intervertebral motion showed much greater mobility in group A than in group B and group C (P < 0.01) but no statistically significant difference between group B and group C (P > 0.05). The range of intervertebral motion was maximal at C(5) - C(6) in group A. The place of maximal motion shifted to the C(4) - C(5) in both group B and group C. C(2) - C(3) showed minimal mobility in all three groups. There was no statistically significant difference (P > 0.05) between male and female. The motion range and degenerative regularity of normal Chinese cervical spine were similar in either sex. The range of total intervertebral motion (from C(2) - C(3) to C(6) - C(7)) had a tendency to decrease rapidly and then became steady with age. For the range of intervertebral motion, the place of maximal motion shifted to the higher level with age from C(5) - C(6) in group A to C(4) - C(5) in group B and group C.


Elevated Aqueous Cytokine Levels in Eyes With Ocular Surface Diseases.
To evaluate cytokine and protein levels in the aqueous humor (AqH) of eyes with ocular surface diseases. Prospective consecutive case series. This study includes 14 patients (aged 62.4 ± 13.7 years) with chronic-phase ocular surface diseases (4 with ocular cicatricial pemphigoid, 5 with chemical burns, 2 with a thermal burn, 2 with Stevens-Johnson syndrome, and 1 with exposure keratitis), 14 matched patients without ocular surface disease (controls with corneal scar), and 30 patients who underwent cataract surgery (healthy controls). AqH samples were collected at the beginning of surgery. AqH levels of cytokines (interleukin [IL]-1α, IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-17A, monocyte chemotactic protein [MCP]-1, interferon [IFN]-α, IFN-γ, macrophage inflammatory protein [MIP]-1α, MIP-1β, P-selectin, E-selectin, soluble-intercellular adhesion molecule [s-ICAM]-1, tumor necrosis factor [TNF]-α, granulocyte-macrophage colony-stimulating factor [GM-CSF], IFN-γ-induced protein [IP]-10) were measured using multiplex beads immunoassays. The levels of IL-6, IL-10, IL-17A, GM-CSF, E-selectin, P-selectin, and s-ICAM in AqH were significantly elevated in eyes with ocular surface diseases (in pg/mL: 1696 ± 804, 4.0 ± 1.0, 24.3 ± 9.8, 26.0 ± 18.3, 5150 ± 1232, 13122 ± 7219, and 7914 ± 2813, respectively), compared to healthy controls (IL-6: 6.36 ± 0.94, P = .001; IL-10: 1.68 ± 0.04, P = .0006; IL-17A: 3.7 ± 0.2, P = .008; GM-CSF: 2.7 ± 0.3, P = .007; E-selectin: 2093 ± 37, P = .0001; P-selectin: 3658 ± 137, P = .0001; sICAM-1: 1397 ± 119, P = .008). The levels of IL-6, IL-17A, E-selectin, and P-selectin in AqH were significantly higher in eyes with ocular surface diseases compared to those with corneal scar (IL-6: 44.1 ± 15.0, P = .0077; IL-17A: 4.1 ± 0.7, P = .034; E-selectin: 2439 ± 302, P = .039; and P-selectin: 5673 ± 1553, P = .017). Multiple AqH cytokine levels were elevated in chronic ocular surface diseases.


Marginal donors: can older donor hearts tolerate prolonged cold ischemic storage?
Both advanced donor age and prolonged ischemic time are significant risk factors for the 1-year mortality. However, its functional consequences have not been fully evaluated in the early-phase after transplantation; even early graft dysfunction is the main determinant of long-term outcome following transplantation. We evaluated in vivo left-ventricular (LV) cardiac and coronary vascular function of old-donor grafts after short and prolonged cold ischemic times in rats 1 h after heart transplantation. The hearts were excised from young donor (3-month-old) or old donor (18-month-old) rats, stored in cold preservation solution for either 1 or 8 h, and heterotopically transplanted. After 1 h of ischemic period, in the old-donor group, LV pressure, maximum pressure development (dP/dt max), time constant of LV pressure decay (τ), LV end-diastolic pressure and coronary blood flow did not differ compared with young donors. However, endothelium-dependent vasodilatation to acetylcholine resulted in a significantly lower response of coronary blood flow in the old-donor group (33 ± 4 vs. 51 ± 15 %, p < 0.05). After 8 h preservation, two of the old-donor hearts showed no mechanical activity upon reperfusion. LV pressure (55 ± 6 vs. 72 ± 5 mmHg, p < 0.05), dP/dt max (899 ± 221 vs. 1530 ± 217 mmHg/s, p < 0.05), coronary blood flow and response to acetylcholine were significantly reduced and τ was increased in the old-donor group in comparison to young controls. During the early-phase after transplantation, the ischemic tolerance of older-donor hearts is reduced after prolonged preservation time and the endothelium is more vulnerable to ischemia/reperfusion.


Association of Disability With Mortality From Opioid Overdose Among US Medicare Adults.
Patients qualifying for Medicare disability have the highest rates of opioid use compared with older Medicare beneficiaries and commercial insurance beneficiaries. Research on opioid overdose deaths in this population can help identify appropriate interventions. To assess the rate of opioid overdose death and to identify its associated risk factors. This cohort study included a 20% national sample of Medicare enrollees aged 21 to 64 years whose initial Medicare entitlement was based on disability and who resided in 50 US states and Washington, DC, in 2012 to 2016. Data analyses were performed from March 15, 2019, through September 23, 2019. Fifty-five chronic or potentially disabling conditions were selected from the Centers for Medicare & Medicaid Services Chronic Disease Data Warehouse. Opioid overdose death rate estimated from Medicare National Death Index linkage data. Among 1 766 790 Medicare enrollees younger than 65 years who qualified for Medicare because of disability, the mean (SD) age was 52.2 (10.2) years, and 866 914 (49.1%) were women. These enrollees represent 14.9% (95% CI, 14.9%-15.0%) of the entire Medicare population and accounted for 80.8% (95% CI, 78.9%-82.7%) of opioid overdose deaths among all Medicare enrollees. Opioid overdose mortality in this population increased from 57.4 per 100 000 (95% CI, 53.9-61.0 per 100 000) in 2012 to 77.6 per 100 000 (95% CI, 73.5-81.8 per 100 000) in 2016. Results from the stepwise logistic regression model revealed that 3 categories of conditions are associated with opioid overdose death: substance abuse, psychiatric diseases, and chronic pain. Among the 11.1% (95% CI, 11.0%-11.2%) of adults with disability who had all 3 conditions, the rate of opioid overdose death was 363.7 per 100 000 (95% CI, 326.7-402.6 per 100 000), which is 23.4 times higher than the rate for individuals with none of the conditions (15.5 per 100 000; 95% CI, 11.6-20.1 per 100 000). This study identifies differences in opioid overdose mortality among subgroups of Medicare enrollees younger than 65 years who qualify for Medicare because of disability. Understanding the heterogeneity of medical and psychiatric conditions associated with opioid use and misuse is key to developing specific, data-driven interventions targeted to each subgroup of high-risk populations.


Diced Cartilage Techniques in Rhinoplasty.
Diced cartilage has been widely applied in rhinoplasty, especially for segmental dorsal augmentation, with favorable outcomes over time. Various techniques were developed to increase both stability and viability of diced cartilage, including wrappings with non-blood material/blood products and changing the shapes of the diced cartilage, while the optimal technique is inconclusive. This article systematically reviews the current strategies of diced cartilage technique for rhinoplasty in the databases and emphasizes different wrapping materials. Free diced cartilage has a potential risk of gathering in certain regions and causing postoperative irregularities. Among non-blood wrapping materials, Surgicel is now rarely used due to severe foreign body reactions. The obvious drawbacks of fascia are donor site morbidity, inadequate quantity, and time-consuming. Although diced cartilage wrapped in tutoplast-processed fascia lata, AlloDerm, or esterified hyaluronic acid has achieved primary encouraging results, the increased inflammation raised controversies regarding their clinical use. Diced cartilage wrapped with blood products, such as platelet-rich fibrin or concentrated growth factor, achieved long-term reliable aesthetic results, and shaved cartilage and ultra-diced cartilage have recently shown satisfactory clinical outcomes, while further study is still needed. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Environmental applications of inorganic-organic clays for recalcitrant organic pollutants removal: Bisphenol A.
Bisphenol-A (BPA) adsorption onto inorganic-organic clays (IOCs) was investigated. For this purpose, IOCs synthesised using octadecyltrimethylammonium bromide (ODTMA, organic modifier) and hydroxy aluminium (Al13, inorganic modifier) were used. Three intercalation methods were employed with varying ODTMA concentration in the synthesis of IOCs. Molecular interactions of clay surfaces with ODTMA and Al13 and their arrangements within the interlayers were determined using Fourier transform infrared spectroscopy (FTIR). Surface area and porous structure of IOCs were determined by applying Brunauer, Emmett, and Teller (BET) method to N2 adsorption-desorption isotherms. Surface area decreased upon ODTMA intercalation while it increased with Al13 pillaring. As a result, BET specific surface area of IOCs was considerably higher than those of organoclays. Initial concentration of BPA, contact time and adsorbent dose significantly affected BPA adsorption into IOCs. Pseudo-second order kinetics model is the best fit for BPA adsorption into IOCs. Both Langmuir and Freundlich adsorption isotherms were applicable for BPA adsorption (R(2)>0.91) for IOCs. Langmuir maximum adsorption capacity for IOCs was as high as 109.89mgg(-1) and it was closely related to the loaded ODTMA amount into the clay. Hydrophobic interactions between long alkyl chains of ODTMA and BPA are responsible for BPA adsorption into IOCs.


Immunocytochemical localization of protein kinase C in developing brain tissue and in primary neuronal cultures.
Antisera to protein kinase C (PKC) have been used to examine the presence and distribution of the enzyme in developing cerebellar cortex of postnatal rat and in cultures of rat sympathetic ganglia. In the cerebellar cortex of 2-,4-, and 6-day old rats, immunostaining was observed in areas of early-forming presynaptic terminals and growth cones. No staining was evident in the cortical proliferative zone. Beginning at 10 days postnatal, nuclear staining, not apparent at earlier stages, was prominent in Purkinje cells. In neuronal cultures of dissociated rat sympathetic ganglia, PKC was immunolocalized in cell bodies and bundles of neuronal processes. Immunoreactivity was particularly striking in growth cones of extending neurites and in axonal varicosities. These results suggest a role for PKC in neuronal growth following cell proliferation and in synaptic function. The appearance of nuclear staining in later developmental stages suggests that the enzyme may be involved in the promotion and maintenance of the differentiated state of neurons.


Lack of effect of influenza vaccine on the pharmacokinetics of antipyrine, alprazolam, paracetamol (acetaminophen) and lorazepam.
31 healthy male (n = 17) and female (n = 14) volunteers, aged 20 to 45 years, were divided into 4 groups and received on 3 separate occasions either: paracetamol (acetaminophen) 650mg intravenously (n = 9); alprazolam 1mg orally (n = 7); antipyrine (phenazone) 1g intravenously (n = 8); or lorazepam 2mg intravenously (n = 7). Doses were administered prior to influenza vaccine (0.5ml, intramuscularly) and at 7 and 21 days post-vaccination. The overall differences among the 3 trials in clearance of antipyrine were of borderline significance (p less than 0.0611), with a trend towards reduced clearance in both of the post-vaccination trials. There were no overall differences observed in the elimination half-life of antipyrine, nor were there significant differences between trials in cumulative urinary excretion or fractional recovery of intact antipyrine, 4-hydroxyantipyrine, norantipyrine, or 3-hydroxymethyl antipyrine. For paracetamol and alprazolam, there were no significant differences among the 3 trials in any of the kinetic variables. The elimination half-life of lorazepam varied significantly among trials, but differences were small and not systematic. Lorazepam clearance did not vary significantly among trials. Thus, clearance of drugs which undergo hepatic conjugative reactions such as glucuronidation and sulphation are unlikely to be affected by the coadministration of influenza vaccine. Furthermore, not all drugs which are biotransformed by hepatic microsomal oxidation necessarily have impaired clearance due to coadministration of influenza vaccine.


The role of plant panallergens in sensitization to natural rubber latex.
Latex allergy represents an increasing occupational problem, mainly among healthcare workers. An association between latex allergy and hypersensitivity to some plant foods, particularly fruits (the latex-fruit syndrome), has been established. Class I chitinases with an N-terminal hevein-like domain from avocado, chestnut, banana and other foods, and latex hevein seem to be the allergens responsible for the cross-reactions involved in the latex-fruit syndrome. The potential role of other latex allergens, such as profilin, Hev b 5, Hev b 7 and beta-1,3-glucanases, in the co-sensitization to latex and plant foods is also discussed.


Biosynthesis of matrix vesicles in epiphyseal cartilage. I. In vivo incorporation of 32P orthophosphate into phospholipids of chondroxyte, membrane, and matrix vesicle fractions.
The in vivo metabolism of 32P orthophosphate into phospholipids of chondroxyte, matrix vesicle, and membrane fractions of chicken epiphyseal cartilage has been studied. Incorporation of radioactive phosphate into the total phospholipid fraction of matrix vesicles was rapid, the labeling of phosphatidylcholine and lysophosphatidylserine being even more rapid in matrix vesicles than in chondrocytes. These findings indicate that matrix vesicles are formed by a rapid, metabolically active process, and are not remnants of dead cells, as had previously been postulated by some workers. The rate of incorporation of 32P orthophosphate into phosphatidylserine and sphingomyelin of matrix vesicles was significantly slower than that of phosphatidylcholine and certain other vesicle phospholipids. These findings are paradoxical because, compared with chondrocytes, matrix vesicles were enriched in phosphatidylserine and sphingomyelin and depleted in phosphatidylcholine. These results indicate that in vesicle formation the rates of degradation of the various phospholipid classes must be markedly different: phosphatidylcholine must be degraded much more rapidly than either phosphatidylserine or sphingomyelin. Support for this comes from previous data which revealed that substantial phospholipase activity is present in epiphyseal cartilage, especially in the zone of hypertrophy where matrix vesicle formation appears to be particularly active.


Comparison of the inhibitory effect of sorbic acid and amphotericin B on Saccharomyces cerevisiae: is growth inhibition dependent on reduced intracellular pH?
The effects of sorbic acid and amphotericin B on the growth and intracellular pH (pHi) of Saccharomyces cerevisiae were studied and compared. Past evidence has suggested that the inhibitory action of sorbic acid on yeast is due to reduction of pHi per se. However, using a novel method to measure pHi in growing cells, little correlation was found between reduced growth rate on exposure to sorbic acid and reduction of pHi. In fact, growth inhibition correlated with an increase in the intracellular ADP/ATP ratio due to increased ATP consumption by the cells. This was partly attributed to the activation of protective mechanisms, such as increased proton pumping by the membrane H(+)-ATPase, which ensured that pHi did not decline when cells were exposed to sorbic acid. Therefore, the available evidence suggested that the inhibitory action of sorbic acid was due to the induction of an energetically expensive protective mechanism that compensated for any disruption of pHi homeostasis but resulted in less available energy for normal growth. In contrast to sorbic acid, with amphotericin B there was a direct correlation between growth inhibition and reduction of pHi due to the uncoupling effect of this compound on the plasma membrane. The inhibitory effect of amphotericin B was consistent with membrane disruption, or 'proton-uncoupling' leading to growth inhibition due to proton influx, decline in pHi and partial dissipation of the proton gradient.


Mitral annulus flattens in ischemic mitral regurgitation: geometric differences between inferior and anterior myocardial infarction: a real-time 3-dimensional echocardiographic study.
New surgical strategies to restore the saddle shape of the mitral annulus are expected to increase annuloplasty effectiveness. Preoperative and postoperative configuration of the curved annulus, however, is difficult to quantify with 2-dimensional echocardiography. We sought to investigate the geometric deformity in the mitral annulus in ischemic mitral regurgitation (MR), comparing inferior and anterior myocardial infarction (MI) with the use of a custom quantitation software system with transthoracic 3-dimensional echocardiography. We performed real-time 3-dimensional echocardiography in 23 patients with ischemic MR attributable to inferior MI or anterior MI and in 10 controls. Three-dimensional data were cropped into 18 radial planes, and we manually marked the annulus in mid systole. Three-dimensional annular images were reconstructed, and annular circumferences, areas, and heights were quantified. Annulus was significantly more dilated and flattened in ischemic MR than in controls and was further deformed in anterior MI as compared with inferior MI (control: circumference 9.9+/-0.7 cm, area 9.6+/-0.5 cm2, height 5.0+/-0.7 mm; inferior MI: circumference 11.5+/-1.2 cm [P<0.01 compared with control], area 11.4+/-2.0 cm2 [P<0.05 compared with control], height 3.5+/-1.6 mm [P<0.05 compared with control]; anterior MI: circumference 14.2+/-2.4 cm [P<0.0001 compared with control, P<0.05 compared with inferior MI], area 13.7+/-2.8 cm2 [P<0.01 compared with control, P<0.05 compared with inferior MI], height 1.7+/-1.5 mm [P<0.0001 compared with control, P<0.05 compared with inferior MI]). Mitral annulus flattens in ischemic MR. Deformity of the mitral annulus was greater in anterior MI group than in the inferior MI group.


Methylnaltrexone for Opioid-Induced Constipation in Children and Adolescents and Young Adults with Progressive Incurable Cancer at the End of Life.
Opioid-induced constipation (OIC) is common among children and adolescents and young adults (AYA) with progressive incurable cancer. Although methylnaltrexone is a successful treatment for OIC in adult cancer patients, no case series has established its safety and efficacy in pediatric cancer patients. The aim of the study was to describe the safety and efficacy of methylnaltrexone use for OIC in children and AYA with progressive incurable cancer at the end of life in the inpatient and outpatient settings. We conducted a retrospective review of medical records of children and AYA with progressive incurable cancer who received methylnaltrexone at our institution from May 2008 to June 2013. Pharmacy data were reviewed for each patient and a chart review was performed for documentation of laxation and side effects. Of the 9 patients (age range: 17 months to 21 years) with progressive incurable cancer who developed OIC, 7 (78%) had laxation after methylnaltrexone administration (0.15 mg/kg/dose). Of these 7 patients, 5 (71%) had laxation with the first dose, and 5 (71%) who responded had a continued response to repeated doses. The longest a patient regularly received methylnaltrexone was 9 months. Of 5 patients with intraabdominal disease, 4 (80%) had laxation. There were no negative side effects in any of the patients. Also, there was no increase in pain either qualitatively or by pain score. Methylnaltrexone appears to be safe and efficacious in treating OIC in children and AYA with progressive incurable cancer. Methylnaltrexone was tolerated in both the inpatient and outpatient settings and with repeated dosing.


Development and evaluation of bioinspired pH-responsive sericin-chitosan-based hydrogel for controlled colonic delivery of PETase: Harnessing PETase-triggered degradation of microplastics.
The gravity of threats posed by microplastic pollution to the environment cannot be overestimated. Being ubiquitous in the living environment, microplastics reach humans through the food chain causing various hazardous effects. Microplastics can be effectively degraded by PETase enzymes. The current study reports, for the first time, a hydrogel-encapsulated, bioinspired colonic delivery of PETase. A free radical polymerization-assisted hydrogel system was synthesized from sericin, chitosan, and acrylic acid using N,N'-methylenebisacrylamide as a crosslinker and ammonium persulfate as an initiator. The hydrogel was characterized with FTIR, PXRD, SEM, and thermal analysis to confirm the development of a stabilized hydrogel system. The hydrogel showed 61 % encapsulation efficiency, maximum swelling, and cumulative PETase release (96 %) at pH 7.4. The mechanism of PETase release exhibited the Higuchi pattern of release with an anomalous transport mechanism. SDS-PAGE analysis confirmed the preservation of the post-release structural integrity of PETase. The released PETase exhibited concentration- and time-dependent degradation of polyethylene terephthalate in vitro. The developed hydrogel system exhibited the intended features of a stimulus-sensitive carrier system that can be efficiently used for the colonic delivery of PETase.


Clostridium septicum gas gangrene following intramuscular infection from an influenza vaccine booster.
We have reported a case of C septicum gas gangrene occurring in the forearm of a patient who had an Influvac influenza booster 48 hours before admission. Despite prompt antibiotic treatment and surgical debridement, the patient became hypotensive and could not be resuscitated.


Dual roles of Pseudomonas aeruginosa AlgE in secretion of the virulence factor alginate and formation of the secretion complex.
AlgE is a monomeric 18-stranded β-barrel protein required for secretion of the extracellular polysaccharide alginate in Pseudomonas aeruginosa. To assess the molecular mechanism of alginate secretion, AlgE was subjected to site-specific and FLAG epitope insertion mutagenesis. Except for β-strands 6 and 10, epitope insertions into the transmembrane β-strands abolished localization of AlgE to the outer membrane. Interestingly, an epitope insertion into β-strand 10 produced alginate and was only detectable in outer membranes isolated from cells grown on solid media. The deletion of nine C-terminal amino acid residues destabilized AlgE. Replacement of amino acids that constitute the highly electropositive pore constriction showed that individual amino acid residues have a specific function in alginate secretion. Two of the triple mutants (K47E+R353A+R459E and R74E+R362A+R459E) severely reduced alginate production. Mutual stability analysis using the algE deletion mutant PDO300ΔalgE(miniCTX) showed the periplasmic alginate biosynthesis proteins AlgK and AlgX were completely destabilized, while the copy number of the inner membrane c-di-GMP receptor Alg44 was reduced. Chromosomal integration of algE restored AlgK, AlgX, and Alg44, providing evidence for a multiprotein complex that spans the cell envelope. Periplasmic turn 4 of AlgE was identified as an important region for maintaining the stability of the putative multiprotein complex.


Systemic and portal haemodynamic changes following triglycyllysine vasopressin plus nitroglycerin administration in patients with hepatitis B-related cirrhosis.
We measured the haemodynamic changes following triglycyllysine vasopressin administration and after addition of nitroglycerin in twelve patients with portal hypertension due to hepatitis B-related cirrhosis. A bolus i.v. injection of triglycyllysine vasopressin at a dose of 2 mg reduced the hepatic venous pressure gradient from 18.5 +/- 3.7 (mean +/- S.D.) to 15.6 +/- 4.0 mmHg, p less than 0.001. However, the cardiac index decreased from 4.8 +/- 1.0 to 3.7 +/- 0.8 l/min m2, p less than 0.001; the heart rate decreased from 79 +/- 15 to 71 +/- 13, p less than 0.01; the right atrial pressure increased from 3.2 +/- 1.9 to 5.3 +/- 2.3 mmHg, p less than 0.001; the mean arterial pressure increased from 92 +/- 13 to 103 +/- 13 mmHg, p less than 0.05; and the systemic vascular resistance rose from 939 +/- 182 to 1367 +/- 310 dyn/s cm-5, p less than 0.001. Furthermore, both mean pulmonary arterial pressure and pulmonary capillary wedge pressure showed a significant increase following triglycyllysine vasopressin administration as compared with baseline values (p less than 0.005). The addition of sublingual nitroglycerin at a dose of 0.6 mg returned all the systemic haemodynamic parameters to baseline levels. On the other hand, nitroglycerin administration caused no further change in the hepatic venous pressure gradient. We concluded that although triglycyllysine vasopressin significantly reduced portal pressure in patients with hepatitis B-related cirrhosis, it produced untoward systemic haemodynamic changes similar to those seen with vasopressin. The addition of nitroglycerin improved the detrimental systemic haemodynamic effects produced by triglycyllysine vasopressin without further reducing the hepatic venous pressure gradient.


Regulating lactate-related immunometabolism and EMT reversal for colorectal cancer liver metastases using shikonin targeted delivery.
There are few effective medications for treating colorectal cancer and liver metastases (CRLM). The interactions among glycolysis, epithelial-mesenchymal transition (EMT), and immune microenvironment contribute to the progression of CRLM. A main glycolytic enzyme pyruvate Kinase M2 (PKM2) is highly expressed in colorectal cancer and CRLM, and thus can be a potential therapeutic target. A therapeutic strategy was proposed and the shikonin-loaded and hyaluronic acid-modified MPDA nanoparticles (SHK@HA-MPDA) were designed for CRLM therapy via PKM2 inhibition for immunometabolic reprogramming. The treatment efficacy was evaluated in various murine models with liver metastasis of colorectal tumor. SHK@HA-MPDA achieved tumor-targeted delivery via hyaluronic acid-mediated binding with the tumor-associated CD44, and efficiently arrested colorectal tumor growth. The inhibition of PKM2 by SHK@HA-MPDA led to the remodeling of the tumor immune microenvironment and reversing EMT by lactate abatement and the suppression of TGFβ signaling; the amount of cytotoxic effector CD8+ T cells was increased while the immunosuppressive MDSCs decreased. The work provided a promising targeted delivery strategy for CRLM treatment by regulating glycolysis, EMT, and anticancer immunity. An immunometabolic strategy for treating colorectal cancer liver metastases using the shikonin-loaded, hyaluronic acid-modified mesoporous polydopamine nanoparticles (SHK@HA-MPDA) via glycolysis inhibition, anticancer immunity activation, and EMT reversal. SHK@HA-MPDA can inhibit cytoplasmic PKM2 and glycolysis of the tumor and reduce lactate flux, and then activate the DCs and remodel the tumor immune microenvironment. The reduced lactate flux can reduce MDSC migration and suppress EMT.


A cocktail of Mycobacterium bovis BCG recombinants expressing the SIV Nef, Env, and Gag antigens induces antibody and cytotoxic responses in mice vaccinated by different mucosal routes.
Recombinant live Mycobacterium bovis BCG strains (rBCG) expressing different human immunodeficiency virus (HIV) or simian immunodeficiency (SIV) antigens could be good candidates for the development of vaccines against AIDS. To develop effective HIV/SIV vaccines, humoral and cellular immune responses directed against multiple antigens may be essential for the control of the infection. In this study we immunized BALB/c mice via different mucosal routes (oral, aerogenic, nasal, and rectal) with a mixture of three rBCG strains expressing, respectively, the entire SIVmac251 Nef protein, and large fragments of the Env and Gag proteins. All routes of immunization studied induced immunoglobulin A (IgA) antibodies against mycobacterial PPD, SIV Env, and SIV Gag antigens in feces and bronchial lavages as well as specific immunoglobulin G (IgG) in serum. Strong, specific cytotoxic responses of splenocytes against Nef, Env, and Gag was observed whatever the mucosal route of immunization. Therefore, mucosal vaccination with a cocktail of rBCG strains induces local, specific IgA, systemic IgG, and systemic CTLs against the three SIV antigens expressed. Rectal and oral routes seemed the most appropriate route of vaccination to be used to protect against SIV infection.


Unraveling the complexity of thermogenic remodeling of white fat reveals potential antiobesity therapies.
Adipose tissue is a complex organ consisting of a mixture of mature adipocytes and stromal vascular cells. It displays a remarkable ability to adapt to environmental and dietary cues by changing its morphology and metabolic capacity. This plasticity is demonstrated by the emergence of interspersed thermogenic beige adipocytes within white depots in response to catecholamines secretion. Coordinated cellular interaction between different cell types within the tissue and a fine-tuned transcriptional program synergistically take place to promote beige remodeling. However, both cell-cell interactions and molecular mechanisms governing beige adipocyte appearance and maintenance are poorly understood. In this and the previous issue of Genes & Development, Shao and colleagues (pp. 1461-1474) and Shan and colleagues (pp. 1333-1338) advance our understanding of these issues and, in doing so, highlight potential therapeutic strategies to combat obesity-associated diseases.


Microvascular anastomosis using a new diode laser system with a contact probe.
The aim of the present study was to demonstrate the usefulness of our newly produced diode laser device (an 830-nm output of 190 mW CW) for microvascular end-to-end anastomoses. In the device, diode laser energy is delivered to a contact ceramic tip with a 200-microns diameter output end. Its wavelength is 830 nm and its power 60 mW. This device is small (30 x 25 x 10 cm), lightweight (gross weight 3kg), and easy to manipulate. Compared with conventional microvascular suture anastomosis (CMSA), the time required for the laser-assisted vascular anastomoses (LAVA) was reduced by almost half. Anastomosed portions of both methods were examined 1, 2, 4, 12, and 24 weeks later for patency and aneurysm formation. The patency rate and incidence of aneurysmal formation were the same in both groups. Histological examination revealed that LAVA is superior to CMSA in the smoothness of the inner surface, less inflammatory reaction, and normal collagen and elastic reorientation. The tensile strength and bursting pressure measured immediately after anastomosis demonstrated no significant difference between the two groups. It was concluded that this diode laser contact method may be clinically useful for microvascular surgery.


Neurodevelopmental Status of Children Aged 6-30 Months With Congenital Heart Disease.
Children with congenital heart diseases (CHD) are considered to be at high-risk for neurodevelopmental delay, but scant Indian data are available. To evaluate the neurodevelopmental status of children with CHD. We enrolled consecutive children aged 6-30 months with echocardiographically-confirmed CHD between June 2013 and January 2014. Children with clinically recognizable genetic syndromes or disorders; visual and/or hearing deficits, and microcephaly; and post-cardiac surgery children were excluded. Development was assessed by Developmental Assessment Scale for Indian Infants (DASII) and Developmental delay defined as Development Quotient (DQ) <70 in either the mental or motor scale. 75 children (53 males) with CHD were enrolled. Acyanotic CHD was seen in 51 children (VSD in 47%), and Tetralogy of Fallot was the commonest cyanotic CHD (25%). Developmental delay was seen in 25% of these children, more in the motor domain (48%) than in mental (12%). Mean motor and mental DQ in acyanotic CHD was 77 and 84, respectively; and 65 and 85, respectively in cyanotic CHD. Mean motor DQ was significantly less than mental DQ in both acyanotic and cyanotic CHD children (P=0.048). Children with CHD are at an increased risk for developmental delay. Periodic surveillance, screening and evaluation should be instituted in them for early identification and appropriate interventions to enhance later academic, behavioral, psycho-social and adaptive function.


Association of increased microvessel density with skeletal extramedullary disease relapse in multiple myeloma patients who have skeletal extramedullary disease at diagnosis.
The aim of the study was to investigate whether microvessel density (MVD) could be associated with skeletal extramedullary disease relapse (skeletal-EMDR) in patients with multiple myeloma (MM) who have skeletal-EMD at diagnosis. Seventy-nine newly diagnosed MM patients who have skeletal-EMD were retrospectively enrolled in this study. The 4-year cumulative incidence of skeletal-EMDR was 35.0%±8.3%. The 4-year probability of overall survival (OS) was 54.0%±7.6%. Multivariate analysis showed that skeletal-EMDR (HR = 4.144; 95% CI: 1.608-10.685; P = 0.003) was independently associated with inferior OS for the MM patients who have skeletal-EMD at diagnosis. The factors associated with skeletal-EMDR were MVD (HR = 3.990, 95%CI:1.136-14.018; P = 0.031), white blood cell (WBC) (HR = 0.262, 95% CI:0.090-0.769; P = 0.015), and the EMD sites involved at onset (HR = 0.263, 95% CI: 0.074-0.937; P = 0.039). The MVD in patients with thoracic and lumbar vertebrae as the involved sites at diagnosis was significantly lower than those with other sites involved (41.59 ± 14.39 vs. 60.82 ± 35.14, P=0.001). Our data suggest that increased MVD could be used to predict skeletal-EMDR, which is associated with inferior survival in patients with MM who have skeletal-EMD at diagnosis.


Comprehensive data on the mechanical properties and biodegradation profile of polylactide composites developed for hard tissue repairs.
Polylactide (PLA), a biopolymer, was reinforced with three fillers (two organic reinforcements and one inorganic filler). The processing technique used to fabricate the composites was the melt-blending technique. The composites and the unreinforced PLA were subjected to microhardness, compression and biodegradation characterisations. Data obtained are presented in this article as raw data. Data from microhardness and compression tests were used to predict the fracture toughness. The biodegradation of the composites was also examined, and the data obtained reported in this article. The data presented in this article allow for a comprehensive understanding of the mechanical behaviour and the biodegradation profile of three composites of PLA with respect to their applications as biodegradable implants. It also helps in the selection of fillers for biopolymers such as PLA.


Intratumoral Heterogeneity in Lung Cancer.
The diagnosis and treatment of lung cancer (LC) is always a challenge. The difficulty in the decision of therapeutic schedule and diagnosis is directly related to intratumoral heterogeneity (ITH) in the progression of LC. It has been proven that most tumors emerge and evolve under the pressure of their living microenvironment, which involves genetic, immunological, metabolic, and therapeutic components. While most research on ITH revealed multiple mechanisms and characteristic, a systemic exposition of ITH in LC is still hard to find. In this review, we describe how ITH in LC develops from the perspective of space and time. We discuss elaborate details and affection of every aspect of ITH in LC and the relationship between them. Based on ITH in LC, we describe a more accurate multidisciplinary therapeutic strategy on LC and provide the newest opinion on the potential approach of LC therapy.


[Redefining time in hospital: the ergonomist's point of view].
Medical and technical evolutions impact on caregivers' working conditions and on patients' treatment. Ergonomics, with the dual aim of improving health and performance, can support these evolutions in an approach focusing on quality of working life.


Estimate of the maximal daily dietary intake of butylated hydroxyanisole and butylated hydroxytoluene in The Netherlands.
The daily dietary intake of the phenolic antioxidants butylated hydroxyanisole (BHA) and/or butylated hydroxytoluene (BHT) was estimated using data obtained from a nationwide dietary record survey carried out in The Netherlands in 1987/1988. The estimates were based on the fat content of selected food categories and their respective maximum permitted levels of BHA and/or BHT. The results indicate that it is unlikely that the current acceptable daily intake for BHA (0-0.05 mg/kg body weight) is surpassed, even in individuals with an extremely high caloric intake, except in extreme cases in 1-6-year-olds. However, it cannot be excluded that the acceptable daily intake for BHT (FAO/WHO: 0-0.125 mg/kg; EEC: 0-0.05 mg/kg) is exceeded in all age and sex groups, but particularly in children aged 1-6 years.


Comparative study of the neuroprotective and nootropic activities of the carboxylate and amide forms of the HLDF-6 peptide in animal models of Alzheimer's disease.
A comparative study of the neuroprotective and nootropic activities of two pharmaceutical substances, the HLDF-6 peptide (HLDF-6-OH) and its amide form (HLDF-6-NH2), was conducted. The study was performed in male rats using two models of a neurodegenerative disorder. Cognitive deficit in rats was induced by injection of the beta-amyloid fragment 25-35 (βA 25-35) into the giant-cell nucleus basalis of Meynert or by coinjection of βA 25-35 and ibotenic acid into the hippocampus. To evaluate cognitive functions in animals, three tests were used: the novel object recognition test, the conditioned passive avoidance task and the Morris maze. Comparative analysis of the data demonstrated that the neuroprotective activity of HLDF-6-NH2, evaluated by improvement of cognitive functions in animals, surpassed that of the native HLDF-6-OH peptide. The greater cognitive/ behavioral effects can be attributed to improved kinetic properties of the amide form of the peptide, such as the character of biodegradation and the half-life time. The effects of HLDF-6-NH2 are comparable to, or exceed, those of the reference compounds. Importantly, HLDF-6-NH2 exerts its effects at much lower doses than the reference compounds.


Respiratory mechanics following chronic cigarette smoke exposure in the Apoe[Formula: see text] mouse model.
Even though cigarette smoking (CS) has been on the decline over the past 50 years, it is still the leading cause of preventable premature death in the United States. Preclinical models have investigated the cardiopulmonary effects of CS exposure (CSE), but the structure-function relationship in the respiratory system has not yet been fully explored. To evaluate these relationships, we exposed female apolipoprotein E-deficient (Apoe[Formula: see text]) mice to mainstream CS ([Formula: see text]) for 5 days/week over 24 weeks with room air as a control (AE, [Formula: see text]). To contextualize the impact of CSE, we also assessed the natural aging effects over 24 weeks of air exposure (baseline, [Formula: see text]). Functional assessments were performed on a small animal mechanical ventilator (flexiVent, SCIREQ), where pressure-volume curves and impedance data at four levels of positive end-expiratory pressure ([Formula: see text]) and with increasing doses of methacholine were collected. Constant phase model parameters ([Formula: see text]: Newtonian resistance, H: coefficient of tissue elastance, and G: coefficient of tissue resistance) were calculated from the impedance data. Perfusion fixed-left lung tissue was utilized for quantification of parenchyma airspace size and tissue thickness, airway wall thickness, and measurements of elastin, cytoplasm + nucleus, fibrin, and collagen content for the parenchyma and airways. Aging caused the lung to become more compliant, with an upward-leftward shift of the pressure-volume curve and a reduction in all constant phase model parameters. This was supported by larger parenchyma airspace sizes, with a reduction in cell cytoplasm + nucleus area. Airway walls became thinner, even though low-density collagen content increased. In contrast, CSE caused a downward-rightward shift of the pressure-volume curve along with an increase in H, G, and hysteresivity ([Formula: see text]). Organ stiffening was accompanied by enhanced airway hyper-responsiveness following methacholine challenge. Structurally, parenchyma airspaces enlarged, as indicated by an increase in equivalent airspace diameter ([Formula: see text]), and the septum thickened with significant deposition of low-density collagen along with an influx of cells. Airway walls thickened due to deposition of both high and low-density collagen, infiltration of cells, and epithelial cell elongation. In all, our data suggest that CSE in female Apoe[Formula: see text] mice reduces respiratory functionality and causes morphological alterations in both central and peripheral airways that results in lung stiffening, compared to AE controls.


A Review of Clinical and Preclinical Studies on Therapeutic Strategies Using Interleukin-12 in Cancer Therapy and the Protective Role of Interleukin-12 in Hematological Recovery in Chemoradiotherapy.
Interleukin-12 (IL-12), a heterodimeric glycoprotein with alpha and ß subunits covalently bonded with a disulfide bond, is a potent anticancer agent. Its action is accomplished through a linkage of the adaptive and innate immune responses. IL-12 can promote the recovery of the hematopoietic system after cancer chemoradiotherapy by stimulating the physiological processes of stem cells, including cell proliferation and differentiation, reconstitution of hematopoietic function, and peripheral blood count recovery. We review therapeutic strategies using IL-12 in clinical studies, including single-agent and combination strategies in hematological tumors and solid tumors, and studies on the protective effects of IL-12 in chemoradiotherapy. This review highlights promising therapeutic strategies based on the anticancer role of IL-12 and the potential protective effects of IL-12 for cancer patients receiving chemoradiotherapy.


Assessment of the physicochemical stability of all-in-one parenteral emulsions for neonates according to USP specifications.
The purpose of this study was to describe the methodology to assess the stability of all-in-one (AIO) parenteral nutrition admixtures, containing glucose, proteins, and lipids, to the standards of U.S. Pharmacopoeia (USP <729>). The influence of calcium and commercially available lipid emulsions and amino acid solutions were also examined. Four batches of 5 AIO admixtures containing calcium were compounded with commercially available lipid emulsions and amino acid solutions. Two of them contained calcium. Their stability was tested under conditions simulating clinical use. All the admixtures were assessed for criteria set by the USP <729>: (1) mean droplet diameter (MDD) and (2) percentage of volume weighted particles with diameter > 5 μm (PFAT5). All admixtures were within the specifications set by the USP with respect to the MDD at 0, 24, and 48 hours, but only those batches lacking calcium met the benchmarks set by the pharmacopoeia, with respect to PFAT5, on the day of preparation. The presence of calcium destabilized the admixtures, while the use of different commercial ingredients altered the admixtures' characteristics. Only 1 batch of the AIO admixtures studied was found to be compliant with USP <729> standards.


NEDD4 activates mitophagy by interacting with LC3 to restrain reactive oxygen species and apoptosis in Apostichopus japonicus challenged with Vibrio splendidus.
Mitophagy, the selective degradation of damaged mitochondria by autophagy, plays a crucial role in the survival of coelomocytes in Apostichopus japonicus following Vibrio splendidus infection by suppressing the generation of reactive oxygen species (ROS) and attenuating cell apoptosis. A recent study revealed that reducing the expression of the neural precursor cell-expressed developmentally downregulated gene 4 (NEDD4), an enzyme 3 (E3) ubiquitin ligase, significantly affects mitochondrial degradation. Prior to the present study, the functional role of NEDD4 in marine invertebrates was largely unexplored. Therefore, we investigated the role of NEDD4 in the activation of mitophagy, modulation of ROS levels, and induction of apoptosis in A. japonicus infected with V. splendidus. The results demonstrated that V. splendidus infection and lipopolysaccharide (LPS) challenge significantly increased the mRNA levels of NEDD4 in A. japonicus coelomocytes, which was consistent with changes in mitophagy under the same conditions. Knockdown of AjNEDD4 using specific small interfering RNAs (siRNAs) impaired mitophagy and caused accumulation of damaged mitochondria, as observed using transmission electron microscopy (TEM) and confocal microscopy. Furthermore, AjNEDD4 was localized to the mitochondria in both coelomocytes and HEK293T cells. Simultaneously, coelomocytes were treated with the inhibitor indole-3-carbinol (I3C) to confirm the regulatory role of AjNEDD4 in mitophagy. The accumulation of AjNEDD4 in the mitochondria and the level of mitophagy decreased. Subsequent investigations demonstrated that AjNEDD4 interacts directly with the microtubule-associated protein light chain 3 (LC3), a key regulator of autophagy and mitophagy, indicating its involvement in the mitophagy pathway. Moreover, AjNEDD4 interference hindered the interaction between AjNEDD4 and LC3, thereby impairing the engulfment and subsequent clearance of damaged mitochondria. Finally, AjNEDD4 interference led to a significant increase in intracellular ROS levels, followed by increased apoptosis. Collectively, these findings suggest that NEDD4 acts as a crucial regulator of mitophagy in A. japonicus and plays a vital role in maintaining cellular homeostasis following V. splendidus infection. NEDD4 suppresses ROS production and subsequent apoptosis by promoting mitophagy, thereby safeguarding the survival of A. japonicus under pathogenic conditions. Further investigation of the mechanisms underlying NEDD4-mediated mitophagy may provide valuable insights into the development of novel strategies for disease control in aquaculture farms.