Cardiac xenotransplantation: a promising way to treat advanced heart failure.
Cardiac xenotransplantation (CXTx) might be a promising approach to bridge the gap between the supply and demand of a donor heart. The survival of cardiac xenograft has been significantly extended in pig-to-nonhuman primate (NHP) CXTx, with records of 195 days and 945 days for orthotropic and heterotopic CXTx, respectively. To present the history of CXTx, we list the reported clinical CXTx, compare pigs and NHPs as sources of hearts, and compare three different kinds of preclinical CXTx models. The application of genetically modified pigs and novel immunosuppressive drugs accelerates the development of CXTx, and we summarize the reported pig-to-NHP CXTx with detailed information. Besides, we discuss the underlining mechanisms and potential preventive strategies of immunological barriers, including hyperacute rejection, acute humoral xenograft rejection, acute cellular xenograft rejection, chronic rejection, coagulation dysfunction, and systemic inflammation. Though intense cellular infiltration in cardiac xenograft has only been documented in a small number of studies, we especially stress the importance of cellular rejection in CXTx, because we believe it is often masked by the rapid and strong humoral response and it may eventually become a more important and common type of xenograft rejection. In addition, we conclude other obstacles as well as possible solutions in CXTx, such as perioperative cardiac xenograft dysfunction, detrimental xenograft overgrowth, and porcine endogenous retroviruses. Finally, we briefly introduce several other approaches that have been proposed to deal with the organ heart shortage crisis, and we firmly believe that CXTx provides the best near-term solution.


Targeting mTOR signaling for cancer therapy.
The mammalian target of rapamycin (mTOR), an atypical serine/threonine kinase, plays a central role in the regulation of cell proliferation, growth, differentiation, migration and survival. Dysregulation of mTOR signaling occurs in diverse human tumours, and can confer higher susceptibility to inhibitors of mTOR. Rapamycin and its derivatives, CCI-779 and RAD001 (designated rapamycins), specifically inhibit the function of mTOR, leading to inactivation of ribosomal S6K1 and inhibition of cap-dependent translation initiation through the 4E-BP1/eIF4E pathway. The overall effect is an accumulation of cells in the G1 phase of the cell-cycle, and potential apoptosis. Preclinical studies indicate that rapamycins are potent inhibitors of the proliferation of numerous tumour cell lines in culture and of murine syngeneic tumour models or human xenografts. RAD001 and CCI-779 are in phase I and II trials, respectively, as anti-cancer agents. These trials have demonstrated promising anti-cancer activity and relatively mild side effects of CCI-779. Emerging results suggest that inhibition of mTOR signaling can be exploited as a potential tumour-selective therapeutic strategy.


On the evaluation of social innovations and social enterprises: Recognizing and integrating two solitudes in the empirical knowledge base.
Social innovation (SI) is billed as a new way to address complex social problems. Interest in SI has intensified rapidly in the last decade, making it an important area of practice for evaluators, but a difficult one to navigate. Learning from developments in SI and evaluation approaches applied in SI contexts is challenging because of 'fuzzy' concepts and silos of activity and knowledge within SI communities. This study presents findings from a systematic review and integration of 41 empirical studies on evaluation in SI contexts. We identify two isolated conversations: one about 'social enterprises' (SEs) and the other about non-SE 'social innovations'. These conversations diverge in key areas, including engagement with evaluation scholarship, and in the reported purposes, approaches and use of evaluation. We identified striking differences with respect to degree of interest in collaborative approaches and facilitation of evaluation use. The findings speak to trends and debates in our field, for example how evaluation might reconcile divergent information needs in multilevel, cross-sectoral collaborations and respond to fluidity and change in innovative settings. Implications for practitioners and commissioners of evaluation include how evaluation is used in different contexts and the voice of evaluators (and the evaluation profession) in these conversations.


Carotid Endarterectomy to Remove Retained Solitaire Stent Retriever inside Carotid Stent after Mechanical Thrombectomy.
Tandem occlusions of the internal carotid artery (ICA) and middle cerebral artery (MCA) occur in up to a third of patients with acute ischemic strokes undergoing endovascular mechanical thrombectomy. Understanding open neurosurgical management of associated complications with this procedure is important. A 67-year-old man with acute onset of left hemiparesis and a tandem right ICA and MCA occlusion. He underwent carotid stent angioplasty of a stenotic ICA, followed by attempted Solitaire stent retrieval of an MCA clot. On withdrawal, the tines of the Solitaire stent lodged inside the Precise carotid stent. The patient was started on aspirin, Plavix, and heparin infusion, and underwent a carotid endarterectomy (CEA) with safe removal of the stents and primary vessel repair. This is the first case reported to date of a Solitaire stent becoming lodged inside a Precise carotid stent, salvaged by CEA with safe removal of the stents and primary vessel repair. We discuss the timing, indication, alternatives, and technical nuances of a CEA in this setting.


205Bi/206Bi cyclotron production from Pb-isotopes for absorption studies in humans.
Pb(p, xn) thick target excitation functions were measured in the energy range 10-38 MeV in order to optimize the production of isotopically pure radiobismuth from natPb, 206Pb, and 207Pb. Additionally, the decay of Po-isotopes from deuteron irradiation of natural bismuth (209Bi) was exploited for radiobismuth production. 205Bi was produced from 206Pb at 20 MeV with only 2% of 206Bi at 4 weeks post irradiation. Bismuth compounds as used in the treatment of peptic ulcer were labeled with 205Bi for absorption studies in animals and subjects.


A new class of sugar analogues for use in the investigation of sugar transport.
D-Mannose derivatives have been synthesised which are crosslinked through their C-4 hydroxyls to propyl-2-amine. Coupling to the amino group gave a fluorodinitrobenzene derivative, a nitroazidophenyl derivative and an azidosalicylamide derivative. Each of these derivatives was shown to have high affinity for the human erythrocyte sugar transport system. The affinity constant for the nitroazidophenyl derivative was not altered by temperature changes. In rat adipocytes treated with insulin, the affinity constants for the derivatives were up to 1000-fold lower than for the parent sugar. In the absence of insulin the affinity constants for the derivatives, but not for D-mannose, were 3-times higher than in insulin-treated cells. By preparation of radiolabelled derivatives we have shown that the compounds are not transported either by erythrocytes or by adipocytes. Thus the crosslinked sugars are good outside-specific analogues.


Schwannomas of the parapharyngeal space and jugular foramen.
Schwannomas or neurilemmomas are among the most common neoplasms occupying the parapharyngeal space, yet only 107 cases have been previously reported. Neurilemmomas involving the jugular foramen are extremely rare. Only 55 cases have been reported in the world literature. The neoplasm occurred in the parapharyngeal space in three of our patients and in the jugular foramen in another patient. Of the tumors located in the parapharyngeal space, the nerve of origin in one of them was the glossopharyngeal, which is extremely rare. Adequate exposure for complete excision of parapharyngeal space tumors is best obtained through an external incision and should not be attempted transorally. In the jugular foramen case, the neoplasm arose from the vagus nerve high in the neck and extended intracranially in a "dumbbell" shape into the posterior cranial fossa. Total removal was successfully accomplished in one stage, by using a subtotal temporal bone resection--upper neck--posterior cranial fossa approach. Surgical removal is the treatment of choice. Schwannomas rarely recur following complete excision.


[Heparin-induced thrombopenia: rapid regression of thrombopenia and thrombosis after plasma exchange. Case report].
Heparin-induced thrombocytopenia with thrombosis is a rare but severe adverse reaction to heparin therapy, whose management is difficult. After heparin withdrawal, the initiation of an alternative anticoagulant therapy, such as recombinant hirudin or danaparoid, is strongly recommended before vitamin K antagonists are effective. Several reports of the efficacity of plasma exchanges in patients with life-threatening thrombosis have been made. We report on a patient with severe aortic thrombosis related to heparin therapy in whom a unique plasma exchange resulted in both dramatic improvement in the platelet count and marked reduction of the thrombosis. This case provides further evidence that plasma exchanges can be useful in the management of heparin-induced thrombocytopenia with thrombosis. They are rapidly efficient and can be used before heparin alternative treatment is effective.


Droplet magnetic-controlled microfluidic chip integrated nucleic acid extraction and amplification for the detection of pathogens and tumor mutation sites.
Traditional nucleic acid extraction and detection is based on open operation, which may cause cross-contamination and aerosol formation. This study developed a droplet magnetic-controlled microfluidic chip integrated nucleic acid extraction, purification and amplification. The reagent is sealed in oil to form a droplet, and the nucleic acid is extracted and purified by controlling the movement of the magnetic beads (MBs) through a permanent magnet, ensuring a closed environment. This chip can automatically extract nucleic acid from multiple samples within 20 min, and can be directly placed in the in situ amplification instrument for amplification without further transfer of nucleic acid, characterized by simple, fast, time-saving and labor-saving. The results showed that the chip was able to detect <10 copies/test SARS-CoV-2 RNA, and EGFR exon 21 L858R mutations were detected in H1975 cells as low as 4 cells. In addition, on the basis of the droplet magnetic-controlled microfluidic chip, we further developed a multi-target detection chip, which used MBs to divide the nucleic acid of the sample into three parts. And the macrolides resistance mutations A2063G and A2064G, and the P1 gene of mycoplasma pneumoniae (MP) were successfully detected in clinical samples by the multi-target detection chip, providing the possibility for future application in the detection of multiple pathogens.


Burden of extensively drug-resistant and pandrug-resistant Gram-negative bacteria at a tertiary-care centre.
The emergence of resistance to multiple antimicrobial agents in Gram-negative bacteria is a significant threat to public health, as it restricts the armamentarium of the clinician against these infections. The aim of this study was to determine the burden of extensively drug-resistant (XDR) and pandrug-resistant (PDR) Gram-negative bacteria at a tertiary-care centre. Antimicrobial susceptibility testing of 1240 clinical isolates of Gram-negative bacteria obtained from various clinical samples during the study period was carried out by the Kirby-Bauer disc diffusion method. Minimum inhibitory concentration of all antibiotics including tigecycline and colistin was determined by Vitek-2 automated susceptibility testing system. Out of 1240 isolates of Gram-negative bacteria, 112 isolates (9%) were resistant to all the antibiotics tested by Kirby-Bauer disc diffusion method. This finding was corroborated by Vitek-2. In addition, Vitek-2 found that 67 isolates were resistant to all antibiotics except tigecycline and colistin. A total of 30 isolates were susceptible to only colistin, and four isolates were susceptible to only tigecycline. It was also found that six isolates (excluding five isolates of Proteus spp.) were resistant to both colistin and tigecycline. Thus, 101 (8.1%) out of 1240 isolates were XDR and 11 isolates (0.9%) were PDR. The findings of this study reveal increased burden of XDR and PDR Gram-negative bacteria in our centre. It also highlights the widespread dissemination of these bacteria in the community. This situation warrants the regular surveillance of antimicrobial resistance of Gram-negative bacteria and implementation of an efficient infection control program.


Glucagon-like peptide-1 plasmid construction and delivery for the treatment of type 2 diabetes.
Glucagon-like peptide-1 (GLP-1) is a 30-amino-acid hormone produced by intestinal L cells. It has been proposed that GLP-1 can be used as a new treatment for type 2 diabetes mellitus because it acts to augment insulin secretion and its effectiveness is maintained in type 2 diabetic patients. Despite its many remarkable advantages as a therapeutic agent for diabetes, GLP-1 is not immediately clinically applicable because of its extremely short half-life. One way to overcome this drawback is GLP1 gene delivery, which enables GLP-1 production in the body. In this study, the effect of GLP1 gene delivery was evaluated both in vitro and in vivo using a new plasmid constructed with a GLP1 (7-37) cDNA. The expression of the GLP1 gene was driven by a SV40 promoter/enhancer. To increase the expression level of GLP-1, nuclear factor kappaB binding sites were introduced. The in vitro results showed expression of GLP-1 and in vitro activity of GLP-1, which is a glucose-dependent insulinotropic action. A single systemic administration of polyethyleneimine/pSIGLP1NFkappaB complex into DIO mice resulted in increasing insulin secretion and decreasing blood glucose levels for a duration longer than 2 weeks.


Presence of an amino acid residue at position 619 required for the function of YidC in Rhodobacter sphaeroides.
YidC, the bacterial homologous protein of Oxa1 and Alb3, could insert membrane proteins into the membrane. Rhodobacter sphaeroides is a kind of photobacteria with abundant intracytoplasmic membranes. In this study, the functions of R. sphaeroides YidC and its C-terminus were investigated in the Escherichia coli YidC gene depletion strain FTL10. The results showed that RS_YidC could complement the growth of the strain FTL10, but the RS_YidC last 5 residues (619-623, KKRKP) deletion mutant could not. Interestingly, the site-directed RS_YidC mutants of any one or all of these 5 residues were still active. The deletion mutant of the last 4 residues and even the last 4 residues deletion mutant with substitution of the Ala or Glu for Lys619 still had sufficient activity to complement the growth of the strain FTL10. These results indicated that the length of the C-terminus of Rs_YidC is more important for its function than the amino acid composition or the charges of it, and the presence of an amino acid residue at position 619 is required for Rs_YidC function in E. coli. Our result also suggests that Rs_YidC may function differently as compared to its homologs.


Reduced MMP-2 activity contributes to cardiac fibrosis in experimental diabetic cardiomyopathy.
To evaluate the regulation of matrix metalloproteinase (MMP)-2 in diabetic cardiomyopathy. Left ventricle (LV) function was determined by a micro-tip catheter in streptozotocin (STZ)-induced diabetic rats, 2 or 6 weeks (w) after STZ-application. LV total collagen, collagen type I and III content were immunohistologically analyzed and quantified by digital image analysis. LV collagen type I, III and MMP-2 mRNA expression was quantified by real-time RT-PCR. LV pro- and active MMP-2 levels were analyzed by zymography; Smad 7, membrane type (MT)1-MMP and tissue inhibitor metalloproteinase (TIMP)-2 protein levels by Western Blot. STZ-induced diabetes was associated with a time-dependent impairment of LV diastolic and systolic function. This was paralleled by a time-dependent increase in LV total collagen content, despite reduced LV collagen type I and III mRNA levels, indicating a role of post-transcriptional/post-translational changes of extracellular matrix regulation. Six weeks (w) after STZ-injection, MMP-2 mRNA expression and pro-MMP-2 levels were 2.7-fold (P < 0.005) and 1.3-fold (P < 0.05) reduced versus controls, respectively, whereas active MMP-2 was decreased to undetectable levels 6 w post-STZ. Concomitantly, Smad 7 and TIMP-2 protein levels were 1.3-fold (P < 0.05) and 10-fold (P < 0.005) increased in diabetics versus controls, respectively, whereas the 45 kDa form of MT1-MMP was undetectable in diabetics. Under STZ-diabetic conditions, cardiac fibrosis is associated with a dysregulation in extracellular matrix degradation. This condition is featured by reduced MMP-2 activity, concomitant with increased Smad 7 and TIMP-2 and decreased MT1-MMP protein expression, which differs from mechanisms involved in dilated and ischemic heart disease.


Hypereosinophilia, neurologic, and gastrointestinal symptoms after bee-pollen ingestion.
A patient developed hypereosinophilia (13,440 cells per cubic millimeter) 6 weeks after beginning the ingestion of bee pollen. Symptoms included generalized malaise, headache, nausea, abdominal pain diarrhea, generalized pruritus, and decreased memory. Evaluation revealed no other known cause for the patient's hypereosinophilia, which resolved after bee-pollen ingestion was stopped. The product contained a mixture of entomophilous and anemophilous pollens to which the patient was skin test positive. An open challenge with the bee pollen later reproduced the presenting symptoms with a concomitant rise of the eosinophil count from 207 to 890 cells per cubic millimeter. The patient has since remained well avoiding bee pollen. This study strongly suggests that hypereosinophilia with attendant pathophysiologic disturbances may be an adverse reaction to bee-pollen ingestion in atopic individuals.


Fine morphological changes in the penicillate nerve endings of human hairy skin during prolonged itching.
Morphological changes in cutaneous nerve endings were investigated electron microscopically in patients suffering from certain kinds of urticaria with associated itching and in normal skin in which wheals and local itching were induced either by application of nettle hairs or by intracutaneous injections of a timothy pollen extract. Skin samples were obtained with a high speed dermal punch without anesthesia from the wheal areas. It was found that some subepidermal free nerve endings derived from non-myelinated nerve fibers (penicillate endings) showed accumulations of extra-cytoplasmic glycogen which was localized in the distended spaces between the axolemma, the Schwann cell membrane and the nerve basement membrane. In some cases, the glycogen was found to be so abundant that it occupied most of the cross sectional area of the ending. No morphological changes were observed in the pappilary endings, in nerve endings of the hairs or in the autonomic terminals. The conducting segments of all cutaneous nerve fibers showed normal morphology. The unusual morphological changes that occur in some penicillate nerve endings during the wheal development indicate that these endings are involved in the skin reaction and therefore they may be the specific end organs that are associated with itch, at least in urticaria.


Exploration of health status, healthcare utilization, and health service expectations among Taiwanese adolescents.
There have been few reports about adolescent experiences with and expectations of health service utilization in an Asian societal setting. The aim of this study is to analyze the use of healthcare services in relation to health status and explore adolescents' preferences for youth-friendly service among Taiwanese high school students. A cross-sectional questionnaire-based survey was conducted on Taiwanese adolescents aged 12–18 years in 2010.We invited participants to rate their health status, report their previous healthcare service use, and rank their health service preferences.We used logistic regression analysis to investigate the association between self-rated health status and healthcare utilization and used nonparametric analysis to compare health service preferences among sociodemographic subgroups. A total of 4,907 students (97.2 % response rate) returned valid questionnaires for analysis. Poor health status and chronic illness were most salient factors independently associated with frequent healthcare service use. Only 40 % of respondents reported having a regular doctor, and pediatrics (57.7 %) was the most commonly identified professional source of medical care. A great majority (86.2 %) of respondents made clinical visits with parents. For characteristics of youth-friendly clinician, the top-ranked items included competency and patience, while having helpful and friendly personnel was highlighted for clinical setting. Family participation is critical in healthcare for adolescents in Asian cultures. Health service use is significantly influenced by health status and chronic illness in the general health insurance system. Understanding these background influences on expectations for healthcare may help to create youth-friendly health services that are more culturally appropriate.


Cytosolic calcium in platelet activation.
Experiments with permeabilised platelets, and with intact platelets loaded with fluorescent Ca2+-indicators, over the past several years have greatly extended our knowledge and understanding of cytosolic Ca2+ as a platelet activator and its interactions with other cytosolic regulators. This article outlines insights, gained from the use of the fluorescent dyes, into maintenance and restoration of basal [Ca2+]i, mechanisms of receptor-mediated Ca2+-mobilisation and quantitation of [Ca2+]i/response relations in intact human platelets.


A microfluidic device for enhanced capture and high activity release of heterogeneous CTCs from whole blood.
Circulating tumor cells (CTCs) are tumor cells that spontaneously detach from the primary focus, and early detection and characterization of CTCs is vital for cancer diagnosis and appropriate treatment. Current methods commonly use EpCAM to capture CTCs, but this results in a loss of information on other CTC subsets (EpCAM-negative cells) due to the heterogeneity of CTCs. Here, we report a novel microfluidic device that integrates the capture and release of heterogeneous CTCs directly from whole blood. A spiral chip was designed for the separation of differently sized cells, and larger CTCs were effectively separated from smaller blood cells with a 98% recovery rate. CD146-containing magnetic beads were used to complement the EpCAM-based CTC capture methods, and the capture efficiency of Fe3O4@Gelatin@CD146/EpCAM increased by 20% over Fe3O4@Gelatin@EpCAM. Finally, MMP-9 was employed to release CTCs with high efficiency and less damage by degrading gelatins on the surface of Fe3O4. The established method was successfully applied to CTC capture and release in a simulated patient's whole blood. The developed method achieved enhanced capture and high activity release of heterogeneous CTCs with less interference by blood cells, which contributes to the early detection and clinical downstream analysis of CTCs.


Web spider's dilemma: a risky move or site dependent growth.
Nephila clavipes inhabiting adjacent habitats showed differences in rate and ratio of growth. Spiders inside the forest fed less, grew slower and stayed smaller than those at the forest edge, a richer environment. The mortality rate in this habitat was about 1% per day, the mortality of vagrant males was even higher. Experimentally fed spiders did not shift their webs readily, even starving individuals stayed much longer than expected. The spiders avoided frequent moves and in a poor site grew more slowly and in smaller size increments. It seems that the costs of small adult size are less than the risks of active foraging.


Plasma IL-1 receptor antagonist levels correlate with the development of non-alcoholic steatohepatitis.
Nonalcoholic steatohepatitis (NASH) is a liver disease characterized by lipid accumulation and inflammation. Here, we aimed to evaluate plasma IL-1Ra as a marker for NASH and to determine whether diagnosis of NASH can be further improved by adding IL-1Ra measurements. Therefore, plasma concentrations of IL-1Ra were measured from 146 subjects of a biopsy-proven NASH cohort with matched controls. NASH patients had higher levels of plasma IL-1Ra compared with patients with steatosis or healthy controls. Our data confirm that IL-1Ra can be a useful tool in the diagnosis of hepatic inflammation and suggest that measuring plasma IL-1Ra levels in addition to ALT will improve the diagnosis for NASH at all stages of the disease.


A spatial map of hepatic mitochondria uncovers functional heterogeneity shaped by nutrient-sensing signaling.
In the liver, mitochondria are exposed to different concentrations of nutrients due to their spatial positioning across the periportal (PP) and pericentral (PC) axis. How these mitochondria sense and integrate these signals to respond and maintain homeostasis is not known. Here, we combined intravital microscopy, spatial proteomics, and functional assessment to investigate mitochondrial heterogeneity in the context of liver zonation. We found that PP and PC mitochondria are morphologically and functionally distinct; beta-oxidation was elevated in PP regions, while lipid synthesis was predominant in the PC mitochondria. In addition, comparative phosphoproteomics revealed spatially distinct patterns of mitochondrial composition and potential regulation via phosphorylation. Acute pharmacological modulation of nutrient sensing through AMPK and mTOR shifted mitochondrial phenotypes in the PP and PC regions, linking nutrient gradients across the lobule and mitochondrial heterogeneity. This study highlights the role of protein phosphorylation in mitochondrial structure, function, and overall homeostasis in hepatic metabolic zonation. These findings have important implications for liver physiology and disease.


When nominal features are marked on verbs: a transcranial magnetic stimulation study.
It has been claimed that verb processing (as opposed to noun processing) is subserved by specific neural circuits in the left prefrontal cortex. In this study, we took advantage of the unusual grammatical characteristics of clitic pronouns in Italian (e.g., lo and la in portalo and portala 'bring it [masculine]/[feminine]', respectively)-the fact that clitics have both nominal and verbal characteristics, to explore the neural correlates of verb and clitic processing. We used repetitive transcranial magnetic stimulation (rTMS) to suppress the excitability of the left prefrontal cortex and to assess its role in producing verb+det+noun and verb+clitic phrases. Results showed an interference effect for both kinds of phrases when stimulation was applied to the left but not to the right prefrontal cortex. However, the interference effect was significantly greater for the verb+clitic than for the verb+det+noun phrases. These findings support the view that clitics increase the morphosyntactic complexity of verbs.


A comparison of approaches to accessing existing biological and chemical relational databases via SPARQL.
Current biological and chemical research is increasingly dependent on the reusability of previously acquired data, which typically come from various sources. Consequently, there is a growing need for database systems and databases stored in them to be interoperable with each other. One of the possible solutions to address this issue is to use systems based on Semantic Web technologies, namely on the Resource Description Framework (RDF) to express data and on the SPARQL query language to retrieve the data. Many existing biological and chemical databases are stored in the form of a relational database (RDB). Converting a relational database into the RDF form and storing it in a native RDF database system may not be desirable in many cases. It may be necessary to preserve the original database form, and having two versions of the same data may not be convenient. A solution may be to use a system mapping the relational database to the RDF form. Such a system keeps data in their original relational form and translates incoming SPARQL queries to equivalent SQL queries, which are evaluated by a relational-database system. This review compares different RDB-to-RDF mapping systems with a primary focus on those that can be used free of charge. In addition, it compares different approaches to expressing RDB-to-RDF mappings. The review shows that these systems represent a viable method providing sufficient performance. Their real-life performance is demonstrated on data and queries coming from the neXtProt project.


Validated stability-indicating methods for determination of cilostazol in the presence of its degradation products according to the ICH guidelines.
Sensitive and selective stability-indicating assay methods (SIAMs) are suggested for the determination of cilostazol (CIL) in the presence of its acid, alkaline and oxidative degradation products. Developing SIAMs is necessary to carry out any stability study. Stress testing of CIL was performed according to the International Conference on Harmonization (ICH) guidelines in order to validate the stability-indicating power of the analytical procedures. Stress testing showed that CIL underwent acid, alkaline and oxidative degradation; on the other hand, it showed stability towards photo- and thermal degradation. Two chromatographic SIAMs were developed, namely HPLC and HPTLC methods. The concentration range and the mean percentage recovery were 1.0-31.0 microg/ml and 99.96+/-0.46 and 0.6-14.0 microg/spot and 99.88+/-1.10 for HPLC and HPTLC methods, respectively. In addition, derivative spectrophotometric methods were developed in order to determine CIL in the presence of its acid degradation product; these were performed by using the third derivative spectra (3D) and the first derivative of the ratio spectra (1DD) methods. The linearity range and the mean percentage recovery were 2.0-34.0 microg/ml and 100.27+/-1.20 for the (3D) method, while they were 2.0-30.0 microg/ml and 99.94+/-1.18 for the (1DD) method. Also, two chemometric-assisted spectrophotometric methods, based on using partial least squares (PLS) and concentration residual augmented classical least squares method (CRACLS), for the determination of CIL were developed. Both methods were applied on zero order spectra of the mixtures of CIL and its acid degradation product, the mean percentage recovery was 100.03+/-1.09 and 99.91+/-1.27 for PLS and CRACLS, respectively. All methods were validated according to the International Conference on Harmonization (ICH) guidelines and applied on bulk powder and pharmaceutical formulations.


Evaluation of 2'-Deoxy-2'-fluoro Antisense Oligonucleotides for Exon Skipping in Duchenne Muscular Dystrophy.
Duchenne muscular dystrophy (DMD) is a severe muscle wasting disorder typically caused by frame-shifting mutations in the DMD gene. Restoration of the reading frame would allow the production of a shorter but partly functional dystrophin protein as seen in Becker muscular dystrophy patients. This can be achieved with antisense oligonucleotides (AONs) that induce skipping of specific exons during pre-mRNA splicing. Different chemical modifications have been developed to improve AON properties. The 2'-deoxy-2'-fluoro (2F) RNA modification is attractive for exon skipping due to its ability to recruit ILF2/3 proteins to the 2F/pre-mRNA duplex, which resulted in enhanced exon skipping in spinal muscular atrophy models. In this study, we examined the effect of two different 2'-substituted AONs (2'-F phosphorothioate (2FPS) and 2'-O-Me phosphorothioate (2OMePS)) on exon skipping in DMD cell and animal models. In human cell cultures, 2FPS AONs showed higher exon skipping levels than their isosequential 2OMePS counterparts. Interestingly, in the mdx mouse model, 2FPS was less efficient than 2OMePS and suggested safety issues as evidenced by increased spleen size and weight loss. Our results do not support a clinical application for 2FPS AON.


The correction of ultrasonic image analysis features for their depth dependence.
Ultrasound has long been used for clinical imaging purposes and for estimating the properties of tissues. More recently image analysis techniques have been used for quantifying the image's texture. These techniques have had limited success due primarily to the dependence of the texture measures on the depth of the sample volume. A simple algorithm has been developed for deriving a correction for this effect; its efficacy is demonstrated on placental data.


Heterogeneity in treatment outcomes and incomplete recovery in first episode psychosis: does one size fit all?
The heterogeneity in recovery outcomes for individuals with First Episode Psychosis (FEP) calls for a strong evidence base to inform practice at an individual level. Between 19-89% of young people with FEP have an incomplete recovery despite gold-standard evidence-based treatments, suggesting current service models, which adopt a 'one-size fits all' approach, may not be addressing the needs of many young people with psychosis. The lack of consistent terminology to define key concepts such as recovery and treatment resistance, the multidimensional nature of these concepts, and common comorbid symptoms are some of the challenges faced by the field in delineating heterogeneity in recovery outcomes. The lack of robust markers for incomplete recovery also results in potential delay in delivering prompt, and effective treatments to individuals at greatest risk. There is a clear need to adopt a stratified approach to care where interventions are targeted at subgroups of patients, and ultimately at the individual level. Novel machine learning, using large, representative data from a range of modalities, may aid in the parsing of heterogeneity, and provide greater precision and sophistication in identifying those on a pathway to incomplete recovery.


RFPNet: Reorganizing feature pyramid networks for medical image segmentation.
Medical image segmentation is a crucial step in clinical treatment planning. However, automatic and accurate medical image segmentation remains a challenging task, owing to the difficulty in data acquisition, the heterogeneity and large variation of the lesion tissue. In order to explore image segmentation tasks in different scenarios, we propose a novel network, called Reorganization Feature Pyramid Network (RFPNet), which uses alternately cascaded Thinned Encoder-Decoder Modules (TEDMs) to construct semantic features in various scales at different levels. The proposed RFPNet is composed of base feature construction module, feature pyramid reorganization module and multi-branch feature decoder module. The first module constructs the multi-scale input features. The second module first reorganizes the multi-level features and then recalibrates the responses between integrated feature channels. The third module weights the results obtained from different decoder branches. Extensive experiments conducted on ISIC2018, LUNA2016, RIM-ONE-r1 and CHAOS datasets show that RFPNet achieves Dice scores of 90.47%, 98.31%, 96.88%, 92.05% (Average between classes) and Jaccard scores of 83.95%, 97.05%, 94.04%, 88.78% (Average between classes). In quantitative analysis, RFPNet outperforms some classical methods as well as state-of-the-art methods. Meanwhile, the visual segmentation results demonstrate that RFPNet can excellently segment target areas from clinical datasets.


What is the state of children's participation in qualitative research on health interventions?: a scoping study.
Children are the focus of numerous health interventions throughout the world, yet the extent of children's meaningful participation in research that informs the adaptation, implementation, and evaluation of health interventions is not known. We examine the type, extent, and meaningfulness of children's participation in research in qualitative health intervention research. A scoping study was conducted of qualitative published research with children (ages 6-11 years) carried out as part of health intervention research. Following Arksey and O'Malley's scoping study methodology and aligned with the PRISMA-ScR guidelines on the reporting of scoping reviews, the authors searched, charted, collated, and summarized the data, and used descriptive and content analysis techniques. Ovid MEDLINE was searched from 1 January 2007 to 2 July 2018 using the keywords children, health intervention, participation, and qualitative research. Study selection and data extraction were carried out by two reviewers independently. Of 14,799 articles screened, 114 met inclusion criteria and were included. The study identified trends in when children were engaged in research (e.g., post-implementation rather than pre-implementation), in topical (e.g., focus on lifestyle interventions to prevent adult disease) and geographical (e.g., high-income countries) focuses, and in qualitative methods used (e.g., focus group). While 78 studies demonstrated meaningful engagement of children according to our criteria, there were substantial reporting gaps and there was an emphasis on older age (rather than experience) as a marker of capability and expertise. Despite evidence of children's meaningful participation, topical, geographical, and methodological gaps were identified, as was the need to strengthen researchers' skills in interpreting and representing children's perspectives and experiences. Based on these findings, the authors present a summary reflective guide to support researchers toward more meaningful child participation in intervention research.


Enhanced early-latency electromagnetic activity in the left premotor cortex is associated with successful phonetic categorization.
Sensory-motor interactions between auditory and articulatory representations in the dorsal auditory processing stream are suggested to contribute to speech perception, especially when bottom-up information alone is insufficient for purely auditory perceptual mechanisms to succeed. Here, we hypothesized that the dorsal stream responds more vigorously to auditory syllables when one is engaged in a phonetic identification/repetition task subsequent to perception compared to passive listening, and that this effect is further augmented when the syllables are embedded in noise. To this end, we recorded magnetoencephalography while twenty subjects listened to speech syllables, with and without noise masking, in four conditions: passive perception; overt repetition; covert repetition; and overt imitation. Compared to passive listening, left-hemispheric N100m equivalent current dipole responses were amplified and shifted posteriorly when perception was followed by covert repetition task. Cortically constrained minimum-norm estimates showed amplified left supramarginal and angylar gyri responses in the covert repetition condition at ~100ms from stimulus onset. Longer-latency responses at ~200ms were amplified in the covert repetition condition in the left angular gyrus and in all three active conditions in the left premotor cortex, with further enhancements when the syllables were embedded in noise. Phonetic categorization accuracy and magnitude of voice pitch change between overt repetition and imitation conditions correlated with left premotor cortex responses at ~100 and ~200ms, respectively. Together, these results suggest that the dorsal stream involvement in speech perception is dependent on perceptual task demands and that phonetic categorization performance is influenced by the left premotor cortex.


The effect of foot type, body length and mass on postural stability.
Poor postural stability is associated with chronic ankle instability. Previous research showed an effect of foot type on postural stability. However, the specific effect of supinated feet remains unclear. Our study aimed to assess the effect of foot type on postural stability, while taking potential confounding effects of body mass and body height into account. Forty-three healthy participants between 18 and 40 years old performed barefooted single leg stance tests with eyes open (EO) and closed (EC) on solid ground, and on a balance board (BB). Foot type was determined from pressure recordings during gait, using the arch index. Ground reaction forces were measured using a force plate. Outcome measures were Center of Pressure Velocity (COPV) divided by body height, and the Horizontal Ground Reaction Force (HGRF) divided by body mass. Generalized Estimating Equations models assessed the differences between supinated, normal and pronated feet during EO, EC and on a BB. During EO an interaction between supinated feet and body mass showed an increase of COPV with 0.03 × 10-2 1/s per kilogram of mass relative to normal feet (p = .03). During EC this interaction was more pronounced with 0.22 × 10-2 1/s increase per kilogram mass (p < .01). The HGRF did not differ between foot types in any of the conditions. Supinated feet have a larger increase in COPV compared to normal feet with increasing mass when standing on solid ground during EO and EC. This indicates that people with supinated feet and a higher mass are less stable during single leg stance. Level 3, associative study.


The cholinergic system modulates kindling and kindling-induced mossy fiber sprouting.
In a previous study, our laboratory demonstrated that the intraventricular infusion of nerve growth factor (NGF) accelerated kindling rates and enhanced mossy fiber sprouting in the absence of noticeable kindling-associated neuronal loss. The purpose of the present study was to investigate whether these NGF effects were mediated via the cholinergic system. This study evaluated the effects of the cholinergic agonist pilocarpine and the cholinergic antagonist scopolamine on kindling rates and kindling-induced mossy fiber sprouting in adult rats. The results showed that pilocarpine accelerated kindling rates and enhanced kindling-induced mossy fiber sprouting in the CA3 region of the hippocampus, whereas scopolamine retarded kindling rates and blocked kindling-induced mossy fiber sprouting in the CA3 and IML regions. These findings suggest that the cholinergic system may contribute to the long-term structural and functional alterations that are characteristic of the kindled state. Moreover, these data provide support for the hypothesis that NGF infusions may mediate kindling and kindling-induced mossy fiber sprouting via regulation of the cholinergic system.


Gunshot injury in the neck with an atypical bullet trajectory.
Gunshot injuries are on a rise in both developed and developing countries, the reason for this may be increased access to firearms. Gunshot injuries to the neck and maxillofacial region are associated with high morbidity and mortality due to the complex anatomy and presence of various vital structures in this region. It is indeed a rare finding that a bullet's trajectory passes through the neck region and does not damage any vital structures. We present one such case of gunshot injury to the neck.


Continuous subcutaneous levodopa-carbidopa for the treatment of advanced Parkinson's disease: is it an improvement on other delivery?
Parkinson's disease (PD) is the second most common neurodegenerative disease and is growing in prevalence and disability. The standard treatment for PD is oral levo-dopa (LD) with carbidopa (CD). As PD progresses, despite higher doses of LD/CD, plasma levels of LD fluctuate, and may be associated with motor fluctuations and dyskinesia. The development of two new subcutaneous preparations of LD/CD (ND0612 and ABBV-951) for the treatment of motor fluctuations in PD is described in detail. Both reduce motor fluctuations and dyskinesia with minor infusion site adverse events. A third subcutaneous preparation, DIZ102, is in early-stage development. The premise for using continuous release LD in advanced PD is that steady state levels of LD will prevent motor fluctuations/dyskinesia, but this is not the whole story, and will limit the benefits of subcutaneous continuous release LD. With its present pump system ND0612 cannot be used as monotherapy, whereas ABBV-951 can be. Having to combine with oral LD/CD will complicate the use of ND0612. Both ND0612 and ABBV-951 only cause modest reductions in OFF time. It is not clear whether these subcutaneous preparations will have more benefits than the intestinal gel, which also reduces OFF time and dyskinesia.


Roles of long non-coding RNAs in angiogenesis-related diseases: Focusing on non-neoplastic aspects.
Angiogenesis is a key process in organ and tissue morphogenesis, as well as growth during human development, and is coordinated by pro- and anti-angiogenic factors. When this balance is affected, the related physiological and pathological changes lead to disease. Long non-coding RNAs (lncRNAs) are an important class of non-coding RNAs that do not encode proteins, but play a dynamic role in regulating gene expression. LncRNAs have been reported to be extensively involved in angiogenesis, particularly tumor angiogenesis. The non-tumor aspects have received relatively little attention and summary, but there is a broad space for research and exploration on lncRNA-targeted angiogenesis in this area. In this review, we focus on lncRNAs in angiogenesis-related diseases other than tumors, such as atherosclerosis, myocardial infarction, stroke, diabetic complications, hypertension, osteoporosis, dermatosis, as well as, endocrine, neurological, and other systemic disorders. Moreover, multiple cell types have been implicated in lncRNA-targeted angiogenesis, but only endothelial cells have attracted widespread attention. Thus, we explore the roles of other cells. Finally, we summarize the potential research directions in the area of lncRNAs and angiogenesis that can be undertaken by combining cutting-edge technology and interdisciplinary research, which will provide new insights into the involvement of lncRNAs in angiogenesis-related diseases.


Right Anterior Parasagittal Craniotomy and Interhemispheric Approach for Resection of a Falcontentorial Junction Meningioma: 2-Dimensional Operative Video.
Falcotentorial meningiomas are rare lesions that occur along the midline at the junction of the falx cerebri and tentorium. When large, these lesions pose a surgical challenge because of the risk of neurologic deficits, including cortical blindness. Delicate structures at risk during resection of these lesions include the sagittal sinus, internal cerebral veins, vein of Galen, straight sinus, torcula, and fornix. Although there are a number of very reasonable approaches, including a supracerebellar infratentorial or occipital interhemispheric, we chose an anterior interhemispheric approach to provide the safest route for accessing the largest portion of the tumor while protecting the deep venous system. After identification of the falcine attachment, the tumor is debulked within the tumor capsule. In this case, the tumor was subtotally removed because of the adherence of a small segment of tumor along the vein of Galen. The patient gave informed consent for surgery and video recording. Institutional review board approval was deemed unnecessary. Used with permission from Barrow Neurological Institute.


Massive cerebral air embolism following percutaneous transhepatic biliary drainage: A case report.
Cerebral air embolism from portal venous gas rarely occurs due to invasive procedures (e.g., endoscopic procedures, liver biopsy, or percutaneous transhepatic biliary drainage) that disrupt the gastrointestinal or hepatobiliary structures. Here, we report a rare case of fatal cerebral air embolism following a series of percutaneous transhepatic biliary drainage tube insertions. A 50-year-old woman with a history of cholecystectomy, liver wedge resection, and hepaticojejunostomy for gallbladder cancer presented with altered mental status 1 week after percutaneous transhepatic biliary drainage tube placement. Extensive cerebral air embolism and acute cerebral infarction. Brain computed tomography and magnetic resonance imaging, hyperbaric oxygen therapy, medical therapy. Despite the use of hyperbaric oxygen therapy and medical treatment including vasopressors, the patient eventually died due to massive systemic air embolism. To date, there have been no reports of cerebral air embolism due to percutaneous transhepatic biliary drainage with pronounced radiologic images. We reviewed previously reported fatal cases associated with endoscopic hepatobiliary procedures and assessed the possible mechanisms and potential causes of air embolism.


Durable Polymer Versus Biodegradable Polymer Drug-Eluting Stents After Percutaneous Coronary Intervention in Patients with Acute Coronary Syndrome: The HOST-REDUCE-POLYTECH-ACS Trial.
Large-scale randomized comparison of drug-eluting stents (DES) based on durable polymer versus biodegradable polymer technology is currently insufficient in patients with acute coronary syndrome (ACS). The present study aimed to prove the noninferiority of the durable polymer DES (DP-DES) compared with the biodegradable polymer DES (BP-DES) in such patients. The HOST-REDUCE-POLYTECH-ACS (Harmonizing Optimal Strategy for Treatment of Coronary Artery Diseases-Comparison of Reduction of Prasugrel Dose or Polymer Technology in ACS Patients) trial is an investigator-initiated, randomized, open-label, adjudicator-blinded, multicenter, noninferiority trial comparing the efficacy and safety of DP-DES and BP-DES in patients with ACS. The primary end point was a patient-oriented composite outcome (a composite of all-cause death, nonfatal myocardial infarction, and any repeat revascularization) at 12 months. The key secondary end point was device-oriented composite outcome (a composite of cardiac death, target-vessel myocardial infarction, or target lesion revascularization) at 12 months. A total of 3413 patients were randomized to receive the DP-DES (1713 patients) and BP-DES (1700 patients). At 12 months, patient-oriented composite outcome occurred in 5.2% in the DP-DES group and 6.4% in the BP-DES group (absolute risk difference, -1.2%; Pnoninferiority<0.001). The key secondary end point, device-oriented composite outcome, occurred less frequently in the DP-DES group (DP-DES vs BP-DES, 2.6% vs 3.9%; hazard ratio, 0.67 [95% CI, 0.46-0.98]; P=0.038), mostly because of a reduction in target lesion revascularization. The rate of spontaneous nonfatal myocardial infarction and stent thrombosis were extremely low, with no significant difference between the 2 groups (0.6% versus 0.8%; P=0.513 and 0.1% versus 0.4%; P=0.174, respectively). In ACS patients receiving percutaneous coronary intervention, DP-DES was noninferior to BP-DES with regard to patient-oriented composite outcomes at 12 months after index percutaneous coronary intervention. Registration: URL: https://wwwclinicaltrials.gov; Unique identifier: NCT02193971.


Can the epicondylar axis be defined accurately in total knee arthroplasty?
As part of a study on rotational alignment of the femoral component in knee replacement, the surgical epicondylar axis was identified in 74 knees and marked with pins. An alternative technique was employed to achieve rotational alignment and in each case the distance between the pins and the cut surface of the posterior condyles was measured. The alignment of the femoral component was measured postoperatively by axial CT scans of the distal femur, allowing an assessment to be made of what would have been achieved if the epicondylar pins had been used for guidance. Only 75% of knees would have been within 3 degrees of the true epicondylar axis using the epicondyles to control rotation. The error range was large (range 6 degrees ER to 11 degrees IR). Our conclusion is that even in experienced hands clinical estimation of the epicondylar axis is inaccurate and should not be relied upon as the sole determinant of femoral rotation.


Wound healing in glaucoma filtering surgery.
Successful glaucoma filtering surgery is characterized by the passage of aqueous humor from the anterior chamber to the subconjunctival space, which results in the formation of a filtering bleb. Aqueous in the subconjunctival space may then exit by multiple pathways. Bleb failure most often results from fibroblast proliferation and subconjunctival fibrosis. Factors associated with an increased risk of bleb failure include youth, aphakia, active anterior segment neovascularization, inflammation, previously failed glaucoma filtering surgery, and, possibly, race. Several surgical and pharmacologic techniques have recently been introduced to enhance success in eyes with poor surgical prognoses. To elucidate the scientific rationale of these methods, we summarize the process of wound healing after glaucoma filtering surgery and describe postoperative clinical and histopathologic features, factors which may affect success, and specific methods to improve surgical success.


Effects of chronic buspirone treatment on nicotine and concurrent nicotine+cocaine self-administration.
Nicotine dependence and cocaine abuse are major public health problems, and most cocaine abusers also smoke cigarettes. An ideal pharmacotherapy would reduce both cigarette smoking and cocaine abuse. Buspirone (Buspar) is a clinically available, non-benzodiazepine anxiolytic medication that acts on serotonin and dopamine systems. In preclinical studies, it reduced cocaine self-administration following both acute and chronic treatment in rhesus monkeys. The present study evaluated the effectiveness of chronic buspirone treatment on self-administration of intravenous (IV) nicotine and IV nicotine+cocaine combinations. Five cocaine-experienced adult rhesus monkeys (Macaca mulatta) were trained to self-administer nicotine or nicotine+cocaine combinations, and food pellets (1 g) during four 1-h daily sessions under a second-order schedule of reinforcement (FR 2 (VR16:S)). Each nicotine+cocaine combination maintained significantly higher levels of drug self-administration than nicotine or cocaine alone (P<0.05-0.001). Buspirone (0.032-0.56 mg/kg/h) was administered IV through one lumen of a double-lumen catheter every 20 min for 23 h each day, for 7-10 consecutive days. Each 7-10-day sequence of buspirone treatment was followed by saline-control treatment for at least 3 days until food- and drug-maintained responding returned to baseline. Buspirone dose-dependently reduced responding maintained by nicotine alone (0.001-0.1 mg/kg/inj; P<0.01) and by nicotine (0.001 or 0.0032 mg/kg/inj)+cocaine combinations (0.0032 mg/kg/inj; P<0.05-0.001) with no significant effects on food-maintained responding. We conclude that buspirone selectively attenuates the reinforcing effects of nicotine alone and nicotine+cocaine polydrug combinations in a nonhuman primate model of drug self-administration.


Study of factors influencing the in vivo methylation of inorganic arsenic in rats.
Previous studies have shown that several factors may influence the methylation of inorganic arsenic by rat liver in vitro (Buchet and Lauwerys, 1985). The present study attempts to assess the relevance of these observations in vivo. Like man, rat inactivates inorganic arsenic by methylation to monomethylarsonic (MMA) and dimethylarsinic (DMA) acids which are excreted in urine along with unchanged inorganic arsenic (Asi). The administration of S-adenosylmethionine alone or in association with reduced (GSH) or oxidized glutathione or acetylcysteine and the increase of hepatic GSH level by butylated hydroxytoluene pretreatment do not stimulate the urinary excretion of the methylated arsenic metabolites following a challenge dose of inorganic arsenic. Conversely a reduction of the hepatic GSH level by phorone pretreatment greatly modifies the metabolism of inorganic arsenic in vivo. A reduction exceeding 90% of the control value leads to a decreased urinary excretion of MMA and DMA and an increased urinary excretion of inorganic arsenic. This is also associated with an increased accumulation of inorganic arsenic in the liver. This suggests that a drastic reduction of GSH level in liver not only impairs the methylation of inorganic arsenic but also impairs its biliary excretion. When GSH depletion is less severe, the total amount of arsenic excreted in urine after a challenge dose of NaAsO2 is not significantly different from that found in unpretreated animals but the proportion of the three metabolic forms is different: MMA is reduced whereas Asi and DMA tend to increase. These changes resemble those found in patients with liver insufficiency (J.P. Buchet, A. Geubel, S. Pauwels, P. Mahieu, and R. Lauwerys (1984). The influence of liver disease on the methylation of arsenite in humans. Arch. Toxicol. 55, 151-154). Long-term pretreatment of rats with CCl4 slightly reduces the amount of MMA and DMA excreted in urine following a challenge dose of inorganic arsenic. This effect may result from a reduction of GSH transferase activity by CCl4. This study demonstrates the important role of liver GSH in the metabolism of inorganic arsenic in vivo.


Primary mediastinal liposarcoma. A report of a case and review of the literature.
A case of primary liposarcoma of the mediastinum with a review of the literature is presented. Liposarcomas may attain a large mass and exhibit various kinds of clinical symptoms according to their localisation. Histological typing is very important in determining the type and extent of therapy. Surgical excision of the tumour is the therapy of choice in all cases. Radiotherapy may be of value for the palliation of unresectable cases.


Effect of inositol and glycine with increasing sodium chloride and constant osmolality on development of rabbit embryos.
Many commercially available culture media have a high sodium content as sodium chloride and sodium bicarbonate. We examined the effects of osmolytes, inositol and glycine, on embryos cultured in synthetic RD medium with a lower concentration of NaCl (93 mM) and a higher concentration of NaCl (116 mM) with media held constant at 270 mosmols. There were no significant effects of either glycine or inositol on embryo growth when the embryos were cultured in RD medium with 93 mM NaCl (P > 0.05). Culture of one-cell embryos for 72 h in RD medium with 93 mM NaCl, 116 mM NaCl and 116 mM NaCl containing 0.56 mM inositol resulted in 77%, 14% and 55% expanded blastocysts, respectively (P < 0.05). Corresponding values for two-cell embryos cultured for 67 h were 84%, 34% and 66% expanded blastocysts (P < 0.05). When one-cell and two-cell embryos were cultured in RD medium with 93 mM NaCl, 116 mM NaCl, 116 mM NaCl plus 1 mM glycine and 116 mM NaCl plus 1 mM glycine and 0.56 mM inositol, expanded blastocysts from one-cell embryos were 79%, 19%, 49%, and 48%, respectively (P < 0.05), and expanded blastocysts from two-cell embryos were 91%, 32%, 52%, and 53%, respectively (P < 0.05). Inositol and glycine presumably behave as osmolytes in providing substantial protection for rabbit one-cell and two-cell embryos cultured in a medium with high NaCl concentration.


Role of insulin-like growth factor-binding proteins in the pathophysiology and tumorigenesis of gastroesophageal cancers.
The insulin family of proteins include insulin-like growth factor binding proteins (IGFBPs) that are classified into two groups based on their differential affinities to IGFs: IGF high-affinity binding proteins (IGFBP1-6) and IGF low-affinity IGFBP-related proteins (IGFBP-rP1-10). IGFBPs interact with many proteins, including their canonical ligands insulin-like growth factor 1 (IGF-I) and IGF-II. Together with insulin-like growth factor 1 (IGF1) receptor (IGF1R), IGF2R, and ligands (IGF1 and IGF2), IGFBPs participate in a complex signaling axis called IGF-IGFR-IGFBP. Numerous studies have demonstrated that the IGF-IGFR-IGFBP axis is relevant in gastrointestinal (GI) and other cancers. The presence of different IGFBPs have been reported in gastrointestinal cancers, including esophageal squamous cell carcinoma (ESCC), esophageal adenocarcinoma (EAD or EAC), and gastric adenocarcinoma (GAD or GAC). A literature-based survey clearly indicates that an urgent need exists for a focused review of the role of IGFBPs in gastrointestinal cancers. The aim of this review is to present the biochemical and molecular characteristics of IGFBPs with an emphasis specifically on the role of these proteins in the pathophysiology and tumorigenesis of gastroesophageal cancers.


In vitro influence of benfluorex and its main metabolites on rat liver microsomal membrane properties.
Benfluorex and its three main metabolites at 30 microM have been shown to inhibit Acyl CoA cholesterol acyl transferase activity in rat liver microsome preparations and to fluidize these membranes, as reflected by a decrease in the lipid order parameter. When drug concentrations were higher (60-200 microM), the compounds differed in their enzymatic inhibition properties but retained the same fluidizing effects. Only the parent compound had a dose-dependent inhibiting effect. These results are discussed with regard to the chemical properties of compounds, in particular their electric charges and their lipophilic characters.


Non-autoimmune combined factor XIII A and B subunit deficiencies in rheumatoid arthritis patients treated with anti-interleukin-6 receptor monoclonal antibody (tocilizumab).
Coagulation factor XIII (FXIII) is a plasma fibrin-stabilizing factor comprising A and B subunits (FXIII-A and FXIII-B, respectively) in the form of a heterotetramer (FXIII-A2B2). A humanized monoclonal antibody to the interleukin-6 receptor (tocilizumab, TCZ) has emerged as an effective treatment for rheumatoid arthritis (RA), because it drastically reduces the inflammation of RA. We previously reported that two TCZ-treated RA patients with acquired FXIII deficiency developed pelvic hemorrhage. Because TCZ treatment had been shown to be related to low FXIII ammonia release activity and FXIII antigen in the two RA cases, we further examined FXIII-related parameters in 36 TCZ-treated RA patients and compared to 29 healthy controls by employing functional and immunologic assays for FXIII. FXIII-A antigen and FXIII amine incorporation and ammonia release activities were significantly lower in the TCZ-treated group than the control group. The TCZ-treated group also showed mildly low FXIII-A2B2 and FXIII-B levels, and their fibrinogen levels were the lower limit of normal. A significant correlation between FXIII-B and fibrinogen was observed in the control and the TCZ groups, suggesting a common metabolic mechanism(s) for these two hepatic proteins. Because the specific activities of FXIII were normal and neither anti-FXIII-A nor anti-FXIII-B antibody was detected, the overall low FXIII level may have resulted from its impaired synthesis under an unbalanced cytokine milieu caused by TCZ treatment. Concomitant deficiencies in multiple hemostatic factors, including FXIII, may lead to an increased risk for hemorrhage in TCZ-treated RA patients.


Information Seeking as A Predictor of Risk Behavior: Testing A Behavior and Risk Information Engagement Model (BRIE).
Prior theoretical models of information seeking have examined its role, primarily, as a determinant of recommended behaviors. In this study, we develop and test the "behavior and risk information engagement" (BRIE) model, which accounts for the reciprocal effects of information seeking from interpersonal and media sources on two risk behaviors - nonmedical marijuana and amphetamine use. We test the model among young Israeli adults (N = 800) using a three-wave prospective observational study (at 6-month intervals). Autoregressive cross-lagged structural equation models showed good fit. Information seeking from interpersonal sources at baseline predicted amphetamine use and marijuana use at 6 months. In both models, seeking drug-related information from interpersonal sources at baseline was also a predictor of seeking information from media sources at 6 months. Information seeking from media sources at 6 months was also a significant predictor of amphetamine use at 12 months. Implications for theory and practice are discussed.


Health, economic and social burden of tobacco in Latin America and the expected gains of fully implementing taxes, plain packaging, advertising bans and smoke-free environments control measures: a modelling study.
To investigate the tobacco-attributable burden on disease, medical costs, productivity losses and informal caregiving; and to estimate the health and economic gains that can be achieved if the main tobacco control measures (raising taxes on tobacco, plain packaging, advertising bans and smoke-free environments) are fully implemented in eight countries that encompass 80% of the Latin American population. Markov probabilistic microsimulation economic model of the natural history, costs and quality of life associated with the main tobacco-related diseases. Model inputs and data on labour productivity, informal caregivers' burden and interventions' effectiveness were obtained through literature review, surveys, civil registrations, vital statistics and hospital databases. Epidemiological and economic data from January to October 2020 were used to populate the model. In these eight countries, smoking is responsible each year for 351 000 deaths, 2.25 million disease events, 12.2 million healthy years of life lost, US$22.8 billion in direct medical costs, US$16.2 billion in lost productivity and US$10.8 billion in caregiver costs. These economic losses represent 1.4% of countries' aggregated gross domestic products. The full implementation and enforcement of the four strategies: taxes, plain packaging, advertising bans and smoke-free environments would avert 271 000, 78 000, 71 000 and 39 000 deaths, respectively, in the next 10 years, and result in US$63.8, US$12.3, US$11.4 and US$5.7 billions in economic gains, respectively, on top of the benefits being achieved today by the current level of implementation of these measures. Smoking represents a substantial burden in Latin America. The full implementation of tobacco control measures could successfully avert deaths and disability, reduce healthcare spending and caregiver and productivity losses, likely resulting in large net economic benefits.


Vascular endothelial growth factors C and D and their VEGFR-2 and 3 receptors in blood and lymphatic vessels in healthy and arthritic synovium.
To localize vascular endothelial growth factor C (VEGF-C) and VEGF-D in synovial specimens in relation to their VEGFR-2 and VEGFR-3 receptors in blood and lymphatic vessels. Immunohistochemical staining and messenger RNA analysis from control and arthritic synovial membrane specimens. Quantitative RT-PCR disclosed that VEGF-C mRNA copy numbers were higher than VEGF-D mRNA copy numbers in the rheumatoid arthritis (RA), osteoarthritis, and control patient groups studied (p < 0.01). Immunohistochemical staining localized VEGF-C to synovial lining cell layer, pericytes, and smooth muscle cells of blood vessels. The number of VEGF-C positive cells was increased in the synovial lining of ankylosing spondylitis (AS) and RA compared to control synovium. However, in contrast to control synovial lining, little if any VEGF-D was detected in AS or RA synovial lining. VEGFR-2 expressing stromal blood vessels, also positive for the vascular endothelial marker PAL-E and the basement membrane marker laminin, were more abundant in RA and AS than in controls. Interestingly, the lymphatic endothelial receptor VEGFR-3 was also expressed in most synovial vessels, especially in the sublining capillaries and venules. VEGF-C is strongly expressed in the hypertrophic synovial lining of arthritic joints, whereas VEGF-D expression is very low in AS and RA. The expression of VEGF-C and VEGF-D in pericytes and smooth muscle cells suggests that these factors may have a role in maintaining vascular homeostasis. The VEGF receptors VEGFR-2 and VEGFR-3 are present in most of the sublining blood vessels. The expression of the lymphatic marker VEGFR-3 in the sublining blood vessels may relate to fluid filtration and/or fenestrations. The relatively few lymphatic vessels along with increased vascular permeability in RA may contribute to the development of tissue edema and joint stiffness.


Retracted: A Neural Network-Based Method for Fast Capture and Tracking of Laser Links between Nonorbiting Platforms.
[This retracts the article DOI: 10.1155/2022/9296770.].


Thromboembolic and bleeding outcomes in patients with atrial fibrillation and valvular heart disease: A descriptive nationwide cohort study.
The risks of thromboembolism and bleeding in patients with atrial fibrillation (AF) and valvular heart disease (VHD) are sparsely described. We described the risk of events in non-anticoagulated and anticoagulated patients with AF and VHD according to the evaluated heart valves, rheumatic or artificial valve classification (EHRA classification), EHRA Type 1 and Type 2 VHD, and within subgroups of EHRA Type 1 and Type 2 VHD. Cohort study of AF patients with coexisting VHD, identified in nationwide Danish registries from 2000 to 2018. Risk of thromboembolism and bleeding after 1 year of follow-up were calculated in each group. We identified 28 770 incident AF patients with VHD. Not surprisingly, we observed the highest risks of thromboembolism in the non-anticoagulated AF patients with EHRA Type 1 and Type 2 VHD (4.9% vs 2.6% and 3.2% vs 1.9%) and the highest risks of bleeding in the anticoagulated AF patients with EHRA Type 1 and Type 2 VHD (6.6% vs 4.3% and 6.1% vs 4.9%). However, within the subgroups of AF patients with EHRA Type 1 and Type 2 VHD, we observed a large proportion of non-anticoagulated patients (32.9%-49.2%), despite a CHA2 DS2 -VASc score of 2≤ in the majority of these patients (81.9%-95.6%). When using data reflecting contemporary clinical practice, we observed markedly different risks of thromboembolism and bleeding in EHRA Type 1 and Type 2 VHD. Additionally, we observed a potential underuse of oral anticoagulation within the subgroups of AF patients with EHRA Type 1 and Type 2 VHD, underlining need for further attention on this patient group.


Sexual dimorphism in myocardial tumor necrosis factor-alpha and cardiac function during endotoxin tolerance.
Preconditioning is injury-induced protection against subsequent injury and may be induced by a variety of stimuli. Both males and females may be preconditioned; however, if females are relatively protected against the initial insult, is their preconditioning threshold higher? We hypothesized that preconditioning injury threshold differences may exist between genders, which may be associated with differences in myocardial inflammatory monokine production. Male and female Sprague-Dawley rats (n=3-5/group) were given intraperitoneal injections of 125 or 500 microg/kg Salmonella typhimurium lipopolysaccharide (ETX) or 0.4 mL normal saline (NS; 154 mmol/L NaCl). After 24 hours, another injection of 500 microg/kg ETX (injury dose) or NS was given, and the animals were incubated an additional 1 or 6 hours. The rats were anesthetized and myocardial function evaluated via the Langendorff perfusion model. Tumor necrosis factor-alpha (TNF-alpha), interleukin-(IL)-1beta, and IL-6 were measured in 1-hour animals via an enzyme-linked immunosorbent assay. Nonpreconditioned rats (PC-) received NS followed by ETX. Preconditioned rats received either 125 microg/kg ETX (PC+125) or 500 microg/kg ETX (PC+500) followed by injury dose ETX. PC+125 and PC+500 males, as well as PC+500 females, were preconditioned and retained cardiac function similar to shams. PC+125 females were not preconditioned with this stimulus and had a decrease in cardiac function similar to PC- rats. Furthermore, PC+125 and PC+500 males, and PC+500 females had decreased release of TNF-alpha after preconditioning, while PC- animals and PC+125 females did not. Males and females can be preconditioned by endotoxin; however the preconditioning threshold is higher in females than males.


Linezolid-induced dyserythropoiesis: chloramphenicol toxicity revisited.
We report here two cases of dyserythropoietic anaemia associated with long-term linezolid use that share striking similarities to chloramphenicol-associated myelotoxicity.


Is cytology capable of adequately grading prostate carcinoma? Matched series of 50 cases comparing cytologic and histologic pattern diagnoses.
A perceived disadvantage of fine-needle aspiration biopsy of the prostate is that aspiration biopsies cannot be correlated with current tissue-grading systems. We compared patterns of cellular arrangement among 50 cases of prostate carcinoma studied in histologic and cytologic specimens obtained simultaneously. Cell patterns were independently scored in a semiquantitative fashion on both histologic and cytologic material, and "predicted Gleason scores" were assigned to the cytologic specimens. When carcinomas were classified as well differentiated (Gleason score 2 to 4), moderately differentiated (Gleason score 5 to 7), or poorly differentiated (Gleason score 8 to 10), there was 84 per cent agreement between histology and cytology. In no instance was there overlap between well-differentiated and poorly differentiated adenocarcinoma in the matched specimens. We believe that it is possible to accurately predict tissue patterns of prostate carcinoma and to estimate Gleason scores in aspiration biopsy specimens.


Computed Tomography-Based Morphometric Analysis of Thoracic Pedicles: An Analysis of 1512 Pedicles and Their Correlation with Sex, Age, Weight and Height.
To examine the morphological features of thoracic pedicles in a Turkish population. This retrospective study was performed with the patients who were underwent thoracic CT for any reason such as trauma or pulmonary disorder. Patient's age, height, weight, and sex were recorded and pedicle length (PL), transverse pedicle diameter (TPD), sagittal pedicle diameter (SPD) and, transverse pedicle angle (TPA) were measured. The right and left pedicles were separately measured. The data obtained was analyzed and compared with other studies in the literature. The highest mean TPA value was 33° at T1, whereas the lowest mean TPA value was 3° at T12. The highest mean PL value was 39.6 mm at T11, whereas the lowest mean PL value was 33.7 mm at T1. The lowest mean SPD value was 7.2 mm, which was measured on T1, and the maximum mean SPD was 11.7 mm on T12. The minimum mean TPD value was 3.8 mm at T5 and the maximum value was 6.2 mm at T1. There was a statistically significant positive correlation between pedicle length and height in all vertebrae. Age had no effect on the morphological features of the thoracic pedicle. In males, PL, TPD, and SPD were higher than females. Compared with other populations, the Turkish population has a smaller pedicle width on sagittal and transverse planes, and their PL and medial angling is similar to those of other populations. Male patients who are taller and overweight have higher pedicle width and length.


Ribonucleotides are signals for mismatch repair of leading-strand replication errors.
To maintain genome stability, mismatch repair of nuclear DNA replication errors must be directed to the nascent strand, likely by DNA ends and PCNA. Here we show that the efficiency of mismatch repair in Saccharomyces cerevisiae is reduced by inactivating RNase H2, which nicks DNA containing ribonucleotides incorporated during replication. In strains encoding mutator polymerases, this reduction is preferential for repair of mismatches made by leading-strand DNA polymerase ε as compared to lagging-strand DNA polymerase δ. The results suggest that RNase-H2-dependent processing of ribonucleotides transiently present in DNA after replication may direct mismatch repair to the continuously replicated nascent leading strand.


IFN-gamma production dominates the early human natural killer cell response to Coxsackievirus infection.
Coxsackieviruses (CV) are important human pathogens that have been implicated in the pathogenesis of several diseases, including myocarditis and pancreatitis. How the human immune system recognizes and controls CV infections is not well understood. Studies in mice suggest that natural killer (NK) cells play a critical role in viral clearance and host survival, but the mechanism(s) by which human NK cells may contribute to the host anti-CV defence has not been investigated. Here we show that CVB3 infection markedly reduces HLA class I cell surface expression but does not increase the expression of the activating NK cell receptor ligands MICA/B and ULBP1-3 on human cells. We also demonstrate that the lowered target cell HLA class I surface expression does not correlate with an increased susceptibility to NK cell-mediated killing. However, NK cells responded with a robust production of interferon gamma (IFN-gamma) when peripheral blood mononuclear cells were cocultured with infected cells. In summary, this study shows that CVB3 interferes with the expression of NK cell receptor ligands on infected cells and indicates that IFN-gamma production, rather than cytotoxicity, marks the early human NK cell response to CVB3 infection.


A Multi-Stage Faster RCNN-Based iSPLInception for Skin Disease Classification Using Novel Optimization.
Nowadays, skin cancer is considered a serious disorder in which early identification and treatment of the disease are essential to ensure the stability of the patients. Several existing skin cancer detection methods are introduced by employing deep learning (DL) to perform skin disease classification. Convolutional neural networks (CNNs) can classify melanoma skin cancer images. But, it suffers from an overfitting problem. Therefore, to overcome this problem and to classify both benign and malignant tumors efficiently, the multi-stage faster RCNN-based iSPLInception (MFRCNN-iSPLI) method is proposed. Then, the test dataset is used for evaluating the proposed model performance. The faster RCNN is employed directly to perform image classification. This may heavily raise computation time and network complications. So, the iSPLInception model is applied in the multi-stage classification. In this, the iSPLInception model is formulated using the Inception-ResNet design. For candidate box deletion, the prairie dog optimization algorithm is utilized. We have utilized two skin disease datasets, namely, ISIC 2019 Skin lesion image classification and the HAM10000 dataset for conducting experimental results. The methods' accuracy, precision, recall, and F1 score values are calculated, and the results are compared with the existing methods such as CNN, hybrid DL, Inception v3, and VGG19. With 95.82% accuracy, 96.85% precision, 96.52% recall, and 0.95% F1 score values, the output analysis of each measure verified the prediction and classification effectiveness of the method.


Sulfur-containing derivatives of amphetamine: thioureas, methanesulfonamides, and trifluoromethanesulfonamides.
N-Methylthiourea, methanesulfonamide, and trifluoromethanesulfonamide derivatives of amphetamine, 4-nitroamphetamine, and 4-aminoamphetamine were synthesized. The N-methylthiourea derivative of amphetamine, as well as the triethylammonium salt of the dithiocarbamate, produced significant depressor effects in normotensive dogs. N,N'-Bis(methanesulfonyl)-1-methyl-2-(4-aminophenyl)ethylamine and N-trifluoromethanesulofonyl-1-methyl-2-phenylethylamine, but not N-methanesulfonyl-1-methyl-2-phenylethylamine and N-trifluoroacetyl-1-methyl-2-(4-trifluoromethanesulfonamido-phenyl))ethylamine, showed significant antiobesity effects in rats.


[Deoxyribonuclease from Corynebacterium diphtheriae: dynamics of synthesis and properties].
Diphtheritic bacteria of PW-8 Massachusetts strain produced into cultural medium only one nucleotidase--endoDNAase. The enzyme was synthesized by the cells during the exponential phase of growth. The DNAase was purified 500-fold and exhibited properties specific to neutral-alkaline DNAases (pH optimum about 7.5, absolute requirements for Me2+, single-step mechanism of substrate hydrolysis). The following properties were typical for the enzyme: absence of distinct specificity to structure of bases surrounding the hydrolyzed bond, formation of 5'-end phosphate groups and slightly higher preference to denatured DNA.


The preservation of quartz grain surface textures following vehicle fire and their use in forensic enquiry.
During a terrorist trial, dispute arose as to whether the temperature produced in a car fire was sufficient to destroy quartz grain surface textures. A series of seven sequential experiments showed that the temperature for quartz surface texture modification/destruction and the production of vugs, vesicles and glassy precipitation ('snowdrifting') occurred at 1200 degrees C under normal atmospheric conditions. By adding a number of man-made and natural substances, it was found that only the presence of salts depressed this modification temperature (to 900 degrees C). Experiments to determine the temperature of fire in a car indicated that the maximum temperature produced under natural conditions (810 degrees C) was insufficient to affect the quartz grain surface textures. These results confirm the use of surface texture analysis of quartz grains recovered from the remains of cars subjected to fire and their use as a forensic indicator.


TRPV4 expression in the renal tubule is necessary for maintaining whole body K+ homeostasis.
The Ca2+-permeable transient receptor potential vanilloid type 4 (TRPV4) channel serves as the sensor of tubular flow, thus being well suited to govern mechanosensitive K+ transport in the distal renal tubule. Here, we directly tested whether the TRPV4 function is significant in affecting K+ balance. We used balance metabolic cage experiments and systemic measurements with different K+ feeding regimens [high (5% K+), regular (0.9% K+), and low (<0.01% K+)] in newly created transgenic mice with selective TRPV4 deletion in the renal tubule (TRPV4fl/fl-Pax8Cre) and their littermate controls (TRPV4fl/fl). Deletion was verified by the absence of TRPV4 protein expression and lack of TRPV4-dependent Ca2+ influx. There were no differences in plasma electrolytes, urinary volume, and K+ levels at baseline. In contrast, plasma K+ levels were significantly elevated in TRPV4fl/fl-Pax8Cre mice on high K+ intake. K+-loaded knockout mice exhibited lower urinary K+ levels than TRPV4fl/fl mice, which was accompanied by higher aldosterone levels by day 7. Moreover, TRPV4fl/fl-Pax8Cre mice had more efficient renal K+ conservation and higher plasma K+ levels in the state of dietary K+ deficiency. H+-K+-ATPase levels were significantly increased in TRPV4fl/fl-Pax8Cre mice on a regular diet and especially on a low-K+ diet, pointing to augmented K+ reabsorption in the collecting duct. Consistently, we found a significantly faster intracellular pH recovery after intracellular acidification, as an index of H+-K+-ATPase activity, in split-opened collecting ducts from TRPV4fl/fl-Pax8Cre mice. In summary, our results demonstrate an indispensable prokaliuretic role of TRPV4 in the renal tubule in controlling K+ balance and urinary K+ excretion during variations in dietary K+ intake. NEW & NOTEWORTHY The mechanoactivated transient receptor potential vanilloid type 4 (TRPV4) channel is expressed in distal tubule segments, where it controls flow-dependent K+ transport. Global TRPV4 deficiency causes impaired adaptation to variations in dietary K+ intake. Here, we demonstrate that renal tubule-specific TRPV4 deletion is sufficient to recapitulate the phenotype by causing antikaliuresis and higher plasma K+ levels in both states of K+ load and deficiency.


Localization by hand-held gamma probe of tumor labeled with antibody "cocktail".
Five groups (n = 4) of congenitally athymic female nude mice bearing subcutaneous implants of CX-1 and/or SW-1116 tumor in the hind limbs received iodine-125 radio-labeled monoclonal antibodies (MoAbs) B72.3 (two groups), 17-1A (two groups), and cocktail (one group) (iodogen method, 50 microCi/10 micrograms/mouse). Daily probe counts were made in duplicate with a hand-held detector over each tumor site and the front leg (background) for 21 days. Animals were sacrificed and appropriate well counts were obtained. All the single MoAb preparations localized well in both tumor cell lines. Uptake of monoclonal antibody 17-1A was similar in the two tumor cell lines, with counts initially high and slowly decreasing over the 21-day period. Tumor/background ratios continued to increase over time, indicating that both tumor lines have similar antigenic expression for the monoclonal antibody 17-1A. This was not the case for monoclonal antibody B72.3, which showed a preferential uptake by the CX-1 tumor, with higher initial counts and prolonged binding of the antibody, giving rise to higher tumor/background ratios. The mixture of monoclonal antibodies B72.3 and 17-1A markedly improved the uptake by the CX-1 tumor cell line but not that by the SW-1116 cell line, where the effect was negative when compared to the uptake of the single MoAb preparations. The use of a monoclonal antibody mixture can enhance targeting of some tumor sites. Due to the heterogeneity of tumor cell lines, even within the same animal, different mixtures of monoclonal antibodies are needed to increase the targeting of tumor.


Electric stimulation fMRI of the perforant pathway to the rat hippocampus.
The hippocampal formation is a brain system that is implicated in learning and memory. The major input to the hippocampus arrives from the entorhinal cortex (EC) to the dentate gyrus (DG) through the perforant path. In the present work, we have investigated the functional properties of this connection by concomitantly applying electrophysiological techniques, deep-brain electric microstimulation and functional magnetic resonance imaging in anesthetized rats. We systematically delivered different current intensities at diverse stimulation frequencies to the perforant path while recording electrophysiological and blood-oxygenation-level-dependent (BOLD) signals. We observed a linear relationship between the current intensity used to stimulate the hippocampal formation and the amplitude and extension of the induced BOLD response. In addition, we found a frequency-dependent spatial pattern of activation. With stimulation protocols and train frequencies used for kindling, the activity strongly spreads ipsilaterally through the hippocampus, DG, subiculum and EC.


Mechanical strain induces phenotypic changes in breast cancer cells and promotes immunosuppression in the tumor microenvironment.
Breast cancer (BCa) proliferates within a complex, three-dimensional microenvironment amid heterogeneous biochemical and biophysical cues. Understanding how mechanical forces within the tumor microenvironment (TME) regulate BCa phenotype is of great interest. We demonstrate that mechanical strain enhanced the proliferation and migration of both estrogen receptor+ and triple-negative (TNBC) human and mouse BCa cells. Furthermore, a critical role for exosomes derived from cells subjected to mechanical strain in these pro-tumorigenic effects was identified. Exosome production by TNBC cells increased upon exposure to oscillatory strain (OS), which correlated with elevated cell proliferation. Using a syngeneic, orthotopic mouse model of TNBC, we identified that preconditioning BCa cells with OS significantly increased tumor growth and myeloid-derived suppressor cells (MDSCs) and M2 macrophages in the TME. This pro-tumorigenic myeloid cell enrichment also correlated with a decrease in CD8+ T cells. An increase in PD-L1+ exosome release from BCa cells following OS supported additive T cell inhibitory functions in the TME. The role of exosomes in MDSC and M2 macrophage was confirmed in vivo by cytotracking fluorescent exosomes, derived from labeled 4T1.2 cells, preconditioned with OS. In addition, in vivo internalization and intratumoral localization of tumor-cell derived exosomes was observed within MDSCs, M2 macrophages, and CD45-negative cell populations following direct injection of fluorescently-labeled exosomes. Our data demonstrate that exposure to mechanical strain promotes invasive and pro-tumorigenic phenotypes in BCa cells, indicating that mechanical strain can impact the growth and proliferation of cancer cell, alter exosome production by BCa, and induce immunosuppression in the TME by dampening anti-tumor immunity.


Polychlorinated biphenyls in serum, liver and adipose tissue of subjects with hepatocellular carcinoma living in a highly polluted area.
Brescia (North Italy) is located in a highly industrialized area with diffuse environmental pollution by polychlorinated biphenyls (PCBs) and with a high incidence of hepatocellular carcinoma (HCC). We aimed to investigate the concentration of PCBs congeners in serum, liver and adipose tissue of patients with HCC living in this area. We enrolled prospectively 101 HCC incident cases. Lipid-adjusted PCBs concentration of single congeners and total PCBs concentration, computed as the sum of 24 PCBs congeners, were measured in the serum of all subjects and in the liver and abdominal fat of 59 HCC cases undergoing surgery. A high correlation was found between PCBs serum and liver (Spearman r=0.79), serum and fat (r=0.91), and liver and fat (r=0.75) concentrations. Serum PCBs concentration in HCC patients did not vary according to demographical, clinical or epidemiological variables, apart from age, as PCBs concentration increased linearly with the subject's age. This study confirms that serum lipid-adjusted PCBs concentration is a valid measure of PCBs body storage and is highly correlated with PCBs concentration detected in other biological samples.


Effects of L-arabinose efflux on λ Red recombination-mediated gene knockout in multiple-antimicrobial-resistant Salmonella enterica serovar Choleraesuis.
In this study, six swine-derived multiple-antimicrobial-resistant (MAR) strains of Salmonella Choleraesuis (S. Choleraesuis) were demonstrated to possess higher efflux pump activity than the wild-type (WT). L-Arabinose, a common inducer for gene expression, modulated S. Choleraesuis efflux pump activity in a dose-dependent manner. At low L-arabinose concentrations, increasing L-arabinose led to a corresponding increase in fluorophore efflux, while at higher L-arabinose concentrations, increasing L-arabinose decreased fluorophore efflux activity. The WT S. Choleraesuis that lacks TolC (ΔtolC), an efflux protein associated with bacterial antibiotic resistance and virulence, was demonstrated to possess a significantly reduced ability to extrude L-arabinose. Further, due to the rapid export of L-arabinose, an efficient method for recombination-mediated gene knockout, the L-arabinose-inducible bacteriophage λ Red recombinase system, has a reduced recombination frequency (~ 12.5%) in clinically isolated MAR Salmonella strains. An increased recombination frequency (up to 60%) can be achieved using a higher concentration of L-arabinose (fivefold) for genetic manipulation and functional analysis for MAR Salmonella using the λ Red system. The study suggests that L-arabinose serves not only as an inducer of the TolC-dependent efflux system but also acts as a competitive substrate of the efflux system. In addition, understanding the TolC-dependent efflux of L-arabinose should facilitate the optimization of L-arabinose induction in strains with high efflux activity.


Seroprevalence and determinants of contagious bovine pleuropneumonia in cattle in Burkina Faso.
Contagious bovine pleuropneumonia (CBPP) is a bacterial disease caused by Mycoplasma mycoides subsp. Mycoides. This disease affects ruminants mainly cattle with respiratory disorders as predominant symptoms. In Burkina Faso, this condition has been considered as enzootic since several years but data on its seroprevalence remains scares. This study aimed to establish the serological prevalence and determinants of CBPP in Burkina Faso in 2017. For this purpose, 3969 serum samples have been collected following a stratified sampling plan based on vaccination coverage in 12 regions, 84 communes, and 210 villages and analyzed using c-ELISA test. Individual seroprevalence was 16.91% (95% CI: 15.74-18.07%), while 84.5% (95% CI: 60.46-80.02%) of communes, chosen as epidemiological units were found positive. The individual prevalence was found to be associated with agro-ecological area (p < 0.05) and a prevalence of 18.70% (95% CI: 16.74-20.66%) was noted in Sahelian areas, while 15.79% (95% CI: 14.34-17.23%) was found in Soudanian areas. The prevalence was also significantly associated with vaccination coverage (p < 0.05) with a prevalence of 13.92% (95% CI: 11.66-16.18%), 19.21% (95% CI: 16.66-20.75%) and 11.61%(95% CI: 9.00-14.23%) for high, moderate, and low vaccination coverage respectively. The individual prevalence was respectively 16.97 (95% CI: 15.56-18.39%) and 17.13% (95% CI: 15.93-18.33%) for female and animals more than 2 years old. According to regions, the highest seroprevalence was found in Plateau Central region (38.18%, 95% CI: 29.1-47.26%), while the lowest was found in Centre-Est Region (7%, 95% CI: 4.5-9.5%). These prevalence data will allow us to adapt the ongoing strategy to control CBPP in Burkina Faso.


Temporal targeting of tumour cells and neovasculature with a nanoscale delivery system.
In the continuing search for effective treatments for cancer, the emerging model is the combination of traditional chemotherapy with anti-angiogenesis agents that inhibit blood vessel growth. However, the implementation of this strategy has faced two major obstacles. First, the long-term shutdown of tumour blood vessels by the anti-angiogenesis agent can prevent the tumour from receiving a therapeutic concentration of the chemotherapy agent. Second, inhibiting blood supply drives the intra-tumoural accumulation of hypoxia-inducible factor-1alpha (HIF1-alpha); overexpression of HIF1-alpha is correlated with increased tumour invasiveness and resistance to chemotherapy. Here we report the disease-driven engineering of a drug delivery system, a 'nanocell', which overcomes these barriers unique to solid tumours. The nanocell comprises a nuclear nanoparticle within an extranuclear pegylated-lipid envelope, and is preferentially taken up by the tumour. The nanocell enables a temporal release of two drugs: the outer envelope first releases an anti-angiogenesis agent, causing a vascular shutdown; the inner nanoparticle, which is trapped inside the tumour, then releases a chemotherapy agent. This focal release within a tumour results in improved therapeutic index with reduced toxicity. The technology can be extended to additional agents, so as to target multiple signalling pathways or distinct tumour compartments, enabling the model of an 'integrative' approach in cancer therapy.


Stearylamine-bearing cationic liposomes kill Leishmania parasites through surface exposed negatively charged phosphatidylserine.
Lipid-associated formulations of antileishmanial agents have proved to be more effective therapies with reduced toxicities. Previous studies from our group and others revealed that liposomes bearing phosphatidylcholine and stearylamine (SA) themselves kill Leishmania and other protozoan parasites in vitro and in vivo, without causing any adverse effect on host. In the present study, we offer detailed insights into the mechanism of action of these liposomes. Mechanism study was carried out using fluorometric, confocal and electron microscopic methods. Herein, we provide evidence for induction of membrane disruption by specific interaction with surface phosphatidylserine (PS) of Leishmania promastigotes and amastigotes, phospholipids normally not found on mammalian cell surface, with SA-containing liposomes. Cell surface PS on different forms of Leishmania facilitated liposome-induced parasite killing. The target selectivity of the liposomes was further proved through inhibition of antileishmanial activity with annexinV, and strong affinity with anionic PS rather than phosphatidic acid-containing liposomes for leishmanicidal activity. SA-bearing liposomes specifically kill Leishmania, but are non-toxic to murine peritoneal macrophages and human erythrocytes.


Dark-adapted thresholds at 10- and 30-deg eccentricities in 10-week-old infants.
We tested the hypothesis that the dark-adapted sensitivity of the near central retina is lower than that of peripheral retina in 10-week-old infants. A spatial two alternative forced-choice psychophysical method was used to estimate the threshold for 2-deg spots presented 10 deg and 30 deg from the center of a screen. Each 10-week-old infant (n = 11) and adult (n = 5) subject was tested at both eccentricities. Every infant's threshold at 10 deg is higher than that at 30 deg. The median difference between the infants' thresholds at 10 deg and 30 deg is about 0.5 log unit. Whereas the scotopic thresholds of adults at 10 deg and 30 deg are virtually identical, the thresholds of infants' near central retina are higher than those in more peripheral retina. Delayed maturation of the near central rod outer segments with consequent lower quantum catch may be the primary determinant of this infantile sensitivity pattern.