Effects of ultrasound-assisted curing on the quality and microstructure of low-sodium beef jerky.
The effects of ultrasonic brine on the physicochemical properties, sensory quality and microstructure of low-sodium beef jerkies were investigated. Compared with control jerky brined in 1.5% NaCl solution, both of the direct reduction and partial replacement of 50% NaCl by KCl decreased the sodium content. Ultrasonic treatment resulted in sharp reduction in shear force. KCl substitution together with ultrasonic treatment caused the highest yield, a* value and water content, and the water activity of all groups were below 0.85. Only the direct reduction of NaCl had negative effects on the sensory quality. KCl substitution combined with ultrasonic treatment was the most suitable for producing low-sodium beef jerkies with high quality, possibly because the mixed salt had equal total salt content to control. Meanwhile, the cavitation and mechanical effects of ultrasound may unfold protein structure, increase myofibrillar fragmentation index and break the muscle fiber, thus improving the quality of beef jerky.


Genomic analyses reveal low mitochondrial and high nuclear diversity in the cyclosporin-producing fungus Tolypocladium inflatum.
Mitochondrial DNA is generally regarded to evolve faster than nuclear DNA in animals, whereas if this is also true in fungi remains unclear. Herein, we annotate the first complete mitochondrial genome (mitogenome) of the cyclosporin-producing fungus Tolypocladium inflatum and report the genome-wide sequence variations among five isolates originating from distantly separated localities. We found that T. inflatum has among the most compact of fungal mitogenomes; its 25 kb DNA molecule encodes all standard fungal mitochondrial genes and harbors only one intron. Transcriptional analyses validated the expression of most conserved genes. We found several uncommon repetitive elements and evidence of gene transfer from the mitochondrion to the nucleus. Phylogenetic analyses confirmed the placement of T. inflatum in the fungal order Hypocreales although there was uncertainty on its family-level affiliation. Comparative genomic analyses among the five isolates identified an overall lower level of intraspecific variation in mitogenomes than in nuclear genomes; however, both the nuclear and mitochondrial genomes revealed similar isolate relationships, not correlating with geographic sources of these isolates. Our study shed new insights into the evolution of the medicinally important ascomycete T. inflatum.


Vacuum and Electromagnetic Fields Treatment to Regenerate a Diffuse Mature Facial Scar Caused by Sulfuric Acid Assault.
Acid attacks are on the rise, and they cause extensive and deep burns, especially on the face. The treatments used to improve the aesthetic, functional and social impact of non-acid scars do not always prove useful for acid scars. This article reports the case of a woman with an extended, mature, acid facial scar, caused by sulfuric acid assault, treated with a recent new procedure that combines the application of vacuum and electromagnetic fields. Before and after the treatment, the aesthetic appearance, and motor function of the face and neck were evaluated, as well as the level of hydration, the amount of sebum, the elasticity, and the pH of the skin. The improvements highlighted after the treatment of the aesthetic and functional characteristics of the face and neck, and of the physical parameters of the skin seemed to indicate that this particular treatment induces tissue regeneration, even in the nerve component. However, it is evident that the rehabilitation pathways of facial wounds and scars must be personalized, and must include continuous psychological support for the patient.


Nutrient and drug effects on cholesterol metabolism in the laying hen.
The laying hen is a highly dynamic model for studies of cholesterol metabolism. Cholesterol biosynthesis takes place primarily in the liver where it is regulated by both diet and drugs. Ovarian cholesterol biosynthesis follows a pattern different from that in liver and is not influenced by dietary fat or cholesterol. The hen responds to high levels of dietary polyunsaturated fat by increasing cholesterol biosynthesis, egg cholesterol deposition, and fecal bile acid excretion. Dietary cholesterol curtails liver cholesterol biosynthesis and may or may not result in increased egg cholesterol deposition and/or increased fecal steroid excretion depending on the level of cholesterol intake. Dietary plant sterols and fiber may moderate egg cholesterol deposition but the conditions under which this takes place are not well defined. D-Thyroxin reduces blood cholesterol, increases blood sterol turnover, and increases egg cholesterol concentration. Triparanol and azasterols prevent desmosterol conversion to cholesterol with resultant appearance of both sterols in blood and eggs. Probucol moderates egg cholesterol deposition by reducing synthesis and/or transfer of the sterol to the egg. Implications for the use of the hen in cholesterol metabolism studies are discussed.


Effects of hemorrhage on umbilical venous return and oxygen delivery in fetal lambs.
The effect of acute hemorrhage (21.7 +/- 1.6%, mean +/- SE of the estimated blood volume) on the distribution of the umbilical venous return (UVR) and oxygen delivery to fetal organs (radionuclide microsphere technique) was studied in 10 chronically catheterized fetal lambs. Hemorrhage decreased UVR and total oxygen delivery to the fetus by 23 and 28%, respectively (P less than 0.001). A greater proportion of the UVR bypassed the liver through the ductus venosus (DV) (P less than 0.01). Consequently, DV blood flow was relatively maintained, and liver blood flow and oxygen delivery derived from the UVR decreased by 41 and 45%, respectively (P less than 0.002). The proportion of cardiac output constituted by DV blood increased by 30% (P less than 0.01). The changes in the distribution of the DV blood were similar to the changes in the distribution of cardiac output. As a result the proportion of organ blood flow and oxygen delivery derived from DV blood increased by 30% in both the upper and lower body organs. These studies show that fetal hemorrhage has a marked effect on the distribution of the UVR to the liver. The preferential distribution of the DV blood to the heart and brain is maintained but not enhanced. Our results indicate that changes in distribution of the DV-derived blood and oxygen following fetal hemorrhage is determined primarily by the responses of the arterial circulation, i.e., circulatory afterload.


Asbestos-induced pulmonary fibrosis is augmented in 8-oxoguanine DNA glycosylase knockout mice.
Asbestos causes asbestosis and malignancies by mechanisms that are not fully established. Alveolar epithelial cell (AEC) injury and repair are crucial determinants of the fibrogenic potential of noxious agents such as asbestos. We previously showed that mitochondrial reactive oxygen species mediate asbestos-induced AEC intrinsic apoptosis and that mitochondrial human 8-oxoguanine-DNA glycosylase 1 (OGG1), a DNA repair enzyme, prevents oxidant-induced AEC apoptosis. We reasoned that OGG1 deficiency augments asbestos-induced pulmonary fibrosis. Compared with intratracheal instillation of PBS (50 μl) or titanium dioxide (100 μg/50 μl), crocidolite or Libby amphibole asbestos (100 μg/50 μl) each augmented pulmonary fibrosis in wild-type C57BL/6J (WT) mice after 3 weeks as assessed by histology, fibrosis score, lung collagen via Sircol, and type 1 collagen expression; these effects persisted at 2 months. Compared with WT mice, Ogg1 homozygous knockout (Ogg1(-/-)) mice exhibit increased pulmonary fibrosis after crocidolite exposure and apoptosis in cells at the bronchoalveolar duct junctions as assessed via cleaved caspase-3 immunostaining. AEC involvement was verified by colocalization studies using surfactant protein C. Asbestos increased endoplasmic reticulum stress in the lungs of WT and Ogg1(-/-) mice. Compared with WT, alveolar type 2 cells isolated from Ogg1(-/-) mice have increased mtDNA damage, reduced mitochondrial aconitase expression, and increased P53 and cleaved caspase-9 expression, and these changes were enhanced 3 weeks after crocidolite exposure. These findings suggest an important role for AEC mtDNA integrity maintained by OGG1 in the pathogenesis of pulmonary fibrosis that may represent a novel therapeutic target.


Nonischemic chest pain following successful percutaneous coronary intervention at a regional referral centre in southern Ontario.
The purpose of this study was to identify factors that predispose individuals to nonischemic chest pain following successful percutaneous coronary intervention (PCI). We prospectively followed, for 6 months, a cohort of 110 patients who underwent PCI. We determined baseline factors associated with post PCI pain via nonlinear mixed model regression; a binomial distribution with logit link was used. The mean age of participants (n = 110) was 64 (SD ± 11.19), 69% were male. The majority had 1 coronary vessel dilated (88%) and a single stent placement (67%). During follow-up, chest pain was prevalent in 54% (95% confidence interval [CI], 44.8-63.7) and 45% (95% CI, 36.0-54.8) of patients, at 3 and 6 months respectively. Less than half of those with chest pain were evaluated for ischemia. Of those evaluated, tests were negative for the majority; 74% and 61% at 3 and 6 months respectively. Higher baseline depression (odds ratio 1.50; 95% CI, 1.13-1.99) scores (Hospital Anxiety Depression Scale) were significantly associated with nonischemic chest pain during follow-up. Higher baseline depression scores were found to be significant risk factors for chest pain of nonischemic origin following successful PCI. A larger study is needed to confirm the predictive value of this and other factors that may contribute to this elusive pain problem. Further research is also required to develop pain management strategies for patients whose chest pain persists in the absence of discernible ischemic causes.


Optimizing utilization of kidneys from deceased donors over 60 years: five-year outcomes after implementation of a combined clinical and histological allocation algorithm.
This 5 year observational multicentre study conducted in the Nord Italian Transplant programme area evaluated outcomes in patients receiving kidneys from donors over 60 years allocated according to a combined clinical and histological algorithm. Low-risk donors 60-69 years without risk factors were allocated to single kidney transplant (LR-SKT) based on clinical criteria. Biopsy was performed in donors over 70 years or 60-69 years with risk factors, allocated to Single (HR-SKT) or Dual kidney transplant (HR-DKT) according to the severity of histological damage. Forty HR-DKTs, 41 HR-SKTs and 234 LR-SKTs were evaluated. Baseline differences generally reflected stratification and allocation criteria. Patient and graft (death censored) survival were 90% and 92% for HR-DKT, 85% and 89% for HR-SKT, 88% and 87% for LR-SKT. The algorithm appeared user-friendly in daily practice and was safe and efficient, as demonstrated by satisfactory outcomes in all groups at 5 years. Clinical criteria performed well in low-risk donors. The excellent outcomes observed in DKTs call for fine-tuning of cut-off scores for allocation to DKT or SKT in high-risk patients.


Relative roles of protein kinase A and protein kinase C in modulation of transient receptor potential vanilloid type 1 receptor responsiveness in rat sensory neurons in vitro and peripheral nociceptors in vivo.
The function of the transient receptor potential vanilloid type 1 capsaicin receptor is subject to modulation by phosphorylation catalyzed by various enzymes including protein kinase C and cAMP-dependent protein kinase. The aim of this study was to compare the significance of the basal and stimulated activity of protein kinase C and cAMP-dependent protein kinase in transient receptor potential vanilloid type 1 receptor responsiveness in the rat in vitro by measurement of the intracellular calcium concentration in cultured trigeminal ganglion neurons and in vivo by determination of the behavioral noxious heat threshold. KT5720, a selective inhibitor of cAMP-dependent protein kinase, reduced the calcium transients induced by capsaicin or the other, much more potent transient receptor potential vanilloid type 1 receptor agonist resiniferatoxin in trigeminal sensory neurons and diminished the drop of the noxious heat threshold (heat allodynia) evoked by intraplantar resiniferatoxin injection. Chelerythrine chloride, a selective inhibitor of protein kinase C, failed to alter either of these responses, although it inhibited the effect of phorbol 12-myristate 13-acetate in the in vitro assay. Staurosporine, a rather nonselective protein kinase inhibitor, failed to reduce the capsaicin- and resiniferatoxin-induced calcium transients but inhibited the resiniferatoxin-evoked heat allodynia. Dibutyryl-cAMP and phorbol 12-myristate 13-acetate, activator(s) of cAMP-dependent protein kinase and protein kinase C, respectively, enhanced the effect of capsaicin in the calcium uptake assay while forskolin, an activator of adenylyl cyclase, augmented that of resiniferatoxin in the heat allodynia model. None of the protein kinase inhibitors or activators altered the calcium transients evoked by high potassium, a nonspecific depolarizing stimulus. It is concluded that basal activity of cAMP-dependent protein kinase, unlike protein kinase C, is involved in the maintenance of transient receptor potential vanilloid type 1 receptor function in somata of trigeminal sensory neurons but stimulation of either cAMP-dependent protein kinase or protein kinase C above the resting level can lead to an enhanced transient receptor potential vanilloid type 1 receptor responsiveness. Similar mechanisms are likely to operate in vivo in peripheral terminals of nociceptive dorsal root ganglion neurons.


Intra-day variation of in vivo prostaglandin F2alpha formation in healthy subjects.
Prostaglandin F(2alpha) (PGF(2alpha)) is a major stable prostaglandin formed in vivo in physiological and pathophysiological situations and has mainly potent vasoconstrictive and pro-inflammatory properties. PGF(2alpha) is now used as an indicator of acute and chronic inflammation in human clinical settings but the extent of daily variation of PGF(2alpha)in vivo in healthy humans is unknown. We quantified levels of the PGF(2alpha) metabolite 15-keto-dihydro-PGF(2alpha) in 10 healthy males and females in spot urine samples during the day (including morning urine sample) and in 24-h urine during the same day. The intra-day coefficient of variation was 20.9%. However, the total mean value of 15-keto-dihydro-PGF(2alpha) in urine collected in the morning did not significantly differ from the mean level of 15-keto-dihydro-PGF(2alpha) in the 24-h urine samples in the 10 subjects. 15-Keto-dihydro-PGF(2alpha) levels in morning urine showed a positive linear correlation with levels of 15-keto-dihydro-PGF(2alpha) in 24-h urine (R=0.72, P<0.05). In conclusion, formation of PGF(2alpha) shows a biological variation within the day in healthy humans which should not be overlooked when planning a clinical study. Single morning urine samples can be used as an alternative to 24-h urine collections for quantification of PGF(2alpha) formation to simplify the sampling regime in larger clinical studies.


Sex markets and sexual opportunity structures of behaviorally bisexual latino men in the urban metropolis of new york city.
Sex markets (the spatially and culturally bounded arenas) that shape bisexual behavior among Latino men have been utilized as a deterministic concept without a sufficient focus on the ability of individuals to make autonomous decisions within such arenas. We nuance the theory of sex markets using the concept of sexual opportunity structures to investigate the ways in which behaviorally bisexual Latino men in the urban metropolis of New York City navigate sexual geographies, cultural meaning systems, sexual scripts, and social institutions to configure their bisexual behaviors. Drawing on 60 in-depth interviews with bisexual Latino men in New York City, we first describe and analyze venues that constitute sexual geographies that facilitate and impede sexual interaction. These also allow for a degree of autonomy in decision-making, as men travel throughout the urban sexual landscape and sometimes even manage to reject norms, such as those imposed by Christian religion. We explore some of the cultural meaning systems and social institutions that regulate sex markets and influence individual decision-making. Secrecy and discretion-regulated by the family, masculinity, migration, and religion-only partially shaped sexual behavior and relationships. These factors create a flux in "equilibrium" in bisexual sex markets in which sociocultural-economic structures constantly interplay with human agency. This article contributes to the literature in identifying dynamic spaces for sexual health interventions that draw on individual agency and empowerment.


A review of masticatory muscle function.
Current reports of the actions of the major muscles of mastication are reviewed for clenching tasks in centric occlusion and eccentric jaw positions, mandibular opening, and unilateral chewing. The relationship of the mandible to the maxillae in the intercuspal position and, consequently, the relationship of the condyle to the fossa of the temporomandibular joint may be controlled by the clinician. Similarly, the number and site of tooth contacts in the intercuspal position and in eccentric jaw positions may be altered. These alterations all produce alteration of muscle activity during functional and parafunctional acts. Some of these alterations are now predictable. Nevertheless, much more study needs to be done before the outcomes of morphologic changes on the muscular system can be accurately predicted. Even now, however, clinicians must understand the nature of clenching and chewing muscle activity and how it can be altered to a more favorable situation for each individual. This review indicates that: The elevator muscles demonstrate maximum activity when even bilateral occlusal contacts occur during clenching in the intercuspal position. The elevator muscles are activated together in the intercuspal zone of tooth contact during chewing when the occlusal contacts are balanced bilaterally in this intercuspal position. Increasing the number of eccentric tooth contacts increases the muscle activity during both chewing and clenching. The medial pterygoid muscle is variable in its action. This variability appears to be related to the chewing pathway and the type of occlusion. Its action is enhanced during laterally directed chewing actions of the mandible. The inferior head of the lateral pterygoid muscle has a reciprocal role with the medial pterygoid muscle during chewing and contributes to forward and lateral bracing of the condyle of the mandible. Although the superior head of the lateral pterygoid muscle appears to be active during mandibular closing, the significance of this finding is not fully understood.


Electrical restitution process in dispersed canine cardiac Purkinje and ventricular cells.
Rate-dependent changes in cardiac action potential duration (APD) have been related to ion accumulation and depletion in restricted extracellular spaces. Isolated cardiac cells lack intercellular clefts and thus are less likely than intact tissue to experience ionic transients immediately outside the plasma membrane. Furthermore, isolated Purkinje cells lack the T tubules found in ventricular tissue and cells, which also can be a site of ion accumulation and depletion. We therefore employed single canine ventricular and Purkinje cells to investigate the contribution of restricted spaces to the electrical restitution process. In both cell types, abrupt changes in cycle length do not alter resting potential, but APD alters with a time constant of approximately 1 min. In addition, both preparations exhibit two components to the electrical restitution process of the APD. The rapid component has a time constant of 66 +/- 17 ms in the ventricle cells and 186 +/- 43 ms in the Purkinje cells. The slower component is both smaller and more variable. These values are similar to those reported in intact canine ventricular and Purkinje tissue. Thus the restitution process of APD, as measured in these isolated cardiac cells, is not markedly dependent on the presence or absence of restricted extracellular spaces.


Essential healthcare services provided to conflict-affected internally displaced populations in low and middle-income countries: A systematic review.
Background: Conflict and violent crises have resulted in over 40 million of internally displaced persons (IDPs). Most affected regions lack access to basic health resources and generally rely on humanitarian support. The objective of this review was to appraise primary health service interventions among conflict-induced internally displaced populations in low and middle income countries between 2000 and 2019. Methods: A systematic review of literature in the following databases: Embase, MEDLINE, PsyArticles, PsycINFO, Scopus, Web of Science, LILAC and CAB Articles, was performed to identify interventions implemented in conflict IDP settings. Results: Initial searches yielded 4578 papers and 30 studies met the inclusion criteria. Descriptivesynthesis analysis was used, and the final selections were assessed using a customized CriticalAppraisal Skills Programme (CASP) checklist. Included papers were from Sub-Saharan Africa, South Asia and the Middle East regions. Most studies were on prevention interventions, especially water treatment and maternal health. Treatment interventions mostly focused on onmalaria and mental health. Only one food and nutrition study with outcome data was identified, indicating limitations in IDP health-related intervention publications. Reported interventions were conducted between one week to five years, and the study qualities were moderate. The most effective interventions were integrated programmes and common challenges were weakstudy methodology and data reporting. Conclusion: Regardless of the intervention types and durations, the services offered were beneficial to the IDPs. More intervention evidence are, however required as shown in gaps around food and nutrition, health education and disease surveillance.


Targeting the Fusion Process of SARS-CoV-2 Infection by Small Molecule Inhibitors.
Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a serious threat to global public health, underscoring the urgency of developing effective therapies. Therapeutics and, more specifically, direct-acting antiviral development are still very much in their infancy. Here, we report that two hepatitis C virus (HCV) fusion inhibitors identified in our previous study, dichlorcyclizine and fluoxazolevir, broadly block human coronavirus entry into various cell types. Both compounds were effective against various human-pathogenic CoVs in multiple assays based on vesicular stomatitis virus (VSV) pseudotyped with the spike protein and spike-mediated syncytium formation. The antiviral effects were confirmed in SARS-CoV-2 infection systems. These compounds were equally effective against recently emerged variants, including the delta variant. Cross-linking experiments and structural modeling suggest that the compounds bind to a hydrophobic pocket near the fusion peptide of S protein, consistent with their potential mechanism of action as fusion inhibitors. In summary, these fusion inhibitors have broad-spectrum antiviral activities and may be promising leads for treatment of SARS-CoV-2, its variants, and other pathogenic CoVs. IMPORTANCE SARS-CoV-2 is an enveloped virus that requires membrane fusion for entry into host cells. Since the fusion process is relatively conserved among enveloped viruses, we tested our HCV fusion inhibitors, dichlorcyclizine and fluoxazolevir, against SARS-CoV-2. We performed in vitro assays and demonstrated their effective antiviral activity against SARS-CoV-2 and its variants. Cross-linking experiments and structural modeling suggest that the compounds bind to a hydrophobic pocket in spike protein to exert their inhibitory effect on the fusion step. These data suggest that both dichlorcyclizine and fluoxazolevir are promising candidates for further development as treatment for SARS-CoV-2.


Alcohol and other factors affecting osteoporosis risk in women.
By about age 35, people reach their peak bone mass. Women lose bone mass slowly after that point until a few years after menopause, when bone mass is lost very rapidly. For middle-aged and older women, healthy bones depend on the development, during younger years, of a strong bone structure and an adequate peak bone mass. There is tenuous evidence that moderate alcohol consumption may protect bone. But human and animal studies clearly indicate that chronic heavy drinking, particularly during adolescence and the young adult years, can dramatically compromise bone quality and may increase osteoporosis risk. Further, research indicates that the effects of heavy alcohol use on bone cannot be reversed, even if alcohol consumption is terminated. Research suggests that in addition to alcohol, other lifestyle factors--such as tobacco use, nutrition, weight-bearing exercise, increased body weight, and hormone replacement therapy--affect bone development and osteoporosis risk in women. However, there has been little examination of how alcohol interacts with these factors to influence bone health.


MiR-301a transcriptionally activated by HIF-2α promotes hypoxia-induced epithelial-mesenchymal transition by targeting TP63 in pancreatic cancer.
Pancreatic cancer (PC) is one of the deadliest cancers worldwide. PC metastasis involves a complex set of events, including epithelial-mesenchymal transition (EMT), that increase tumor cell invasiveness. Recent evidence has shown that hypoxia is a major EMT regulator in pancreatic cancer cells and facilitates metastasis; however, the mechanisms remain elusive. To investigate the role of miR-301a in hypoxia-induced EMT in PC cells. Real-time PCR and Western blot analysis were used to detect the expression of miR-301a and EMT markers in PDAC cells cultured in hypoxic and normoxic conditions. Western blot analysis was used to detect the expression of EMT markers in PDAC cells with miR-301a overexpression. Wound healing assay and Transwell assay were used to detect the migration capabilities of PDAC cells with miR-301a overexpression and knockout. Luciferase assay was used to detect the miR-301a promoter and the 3' untranslated region activity of TP63. Orthotopic PC mouse models were used to study the role of miR-301a in metastasis of PDAC cells in vivo. In situ hybridization assay was used to detect the expression of miR-301a in PDAC patient samples (adjacent paratumor and paired tumor tissues). . Hypoxic environment could directly promote the EMT of PC cells. The expression level of miR-301a was increased in a HIF2α dependent manner in hypoxia-cultured CFPAC-1 and BxPC-3 cells. Overexpression of miR-301a enhanced the hypoxia-induced EMT of PC cells, while knocking out miR-301a result in the suppression of hypoxia-induced EMT. TP63 was a direct target of miR-301a and involved in the metastatic process of PC cells. Furthermore, miR-301a upregulation facilitated PDAC distant metastasis and lymph node metastasis in vivo. Additionally, miR-301a overexpression was indicative of aggressive clinicopathological behaviors and poor prognosis. The newly identified HIF-2α-miR301a-TP63 signaling pathway may play a crucial role in hypoxia-induced EMT in PDAC cells.


Tadalafil Nanoemulsion Mists for Treatment of Pediatric Pulmonary Hypertension via Nebulization.
Oral tadalafil (TD) proved promising in treating pediatric pulmonary arterial hypertension (PAH). However, to ensure higher efficacy and reduce the systemic side effects, targeted delivery to the lungs through nebulization was proposed as an alternative approach. This poorly soluble drug was previously dissolved in nanoemulsions (NEs). However, the formulations could not resist aqueous dilution, which precluded its dilution with saline for nebulization. Thus, the current study aimed to modify the previous systems into dilutable TD-NEs and assess their suitability for a pulmonary application. In this regard, screening of various excipients was conducted to optimize the former systems; different formulations were selected and characterized in terms of physicochemical properties, nebulization performance, stability following sterilization, and biocompatibility. Results showed that the optimal system comprised of Capmul-MCM-EP:Labrafac-lipophile (1:1) (w/w) as oil, Labrasol:Poloxamer-407 (2:1) (w/w) as surfactant mixture (Smix) and water. The optimum formulation P2TD resisted aqueous dilution, exhibited reasonable drug loading (2.45 mg/mL) and globule size (25.04 nm), acceptable pH and viscosity for pulmonary administration, and could be aerosolized using a jet nebulizer. Moreover, P2TD demonstrated stability following sterilization and a favorable safety profile confirmed by both in-vitro and in-vivo toxicity studies. These favorable findings make P2TD promising for the treatment of pediatric PAH.


[Purity criteria for Bdellovibrio bacteriovorus cultures].
Two-component cultures of Bdellovibrio bacteriovorus, a bacterial parasite, are not always pure; sometimes they contain microbial forms, different from the host and parasite, which cannot be isolated by conventional techniques of inoculation on solid growth media. The only way to isolate them is to apply techniques used for the reversion of L-forms of bacteria. The isolated microorganisms have been identified. The criteria of purity were established for two-component cultures consisting of the host and parasite.


Human exposure to PCDDs and their precursors from heron and tern eggs in the Yangtze River Delta indicate PCP origin.
Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are highly toxic to humans and wildlife. In the present study, PCDD/Fs were analyzed in the eggs of whiskered terns (Chlidonias hybrida), and genetically identified eggs from black-crowned night herons (Nycticorax nycticorax) sampled from two lakes in the Yangtze River Delta area, China. The median toxic equivalent (TEQ) of PCDD/Fs were 280 (range: 95-1500) and 400 (range: 220-1100) pg TEQ g-1 lw (WHO, 1998 for birds) in the eggs of black-crowned night heron and whiskered tern, respectively. Compared to known sources, concentrations of PCDDs relative to the sum of PCDD/Fs in bird eggs, demonstrated high abundance of octachlorodibenzo-p-dioxin (OCDD), 1,2,3,4,6,7,8-heptaCDD and 1,2,3,6,7,8-hexaCDD indicating pentachlorophenol (PCP), and/or sodium pentachlorophenolate (Na-PCP) as significant sources of the PCDD/Fs. The presence of polychlorinated diphenyl ethers (PCDEs), hydroxylated and methoxylated polychlorinated diphenyl ethers (OH- and MeO-PCDEs, known impurities in PCP products), corroborates this hypothesis. Further, significant correlations were found between the predominant congener CDE-206, 3'-OH-CDE-207, 2'-MeO-CDE-206 and OCDD, indicating a common origin. Eggs from the two lakes are sometimes used for human consumption. The WHO health-based tolerable intake of PCDD/Fs is exceeded if eggs from the two lakes are consumed regularly on a weekly basis, particularly for children. The TEQs extensively exceed maximum levels for PCDD/Fs in hen eggs and egg products according to EU legislation (2.5 pg TEQ g-1lw). The results suggest immediate action should be taken to manage the contamination, and further studies evaluating the impacts of egg consumption from wild birds in China. Likewise, studies on dioxins and other POPs in common eggs need to be initiated around China.


The active metabolite of spleen tyrosine kinase inhibitor fostamatinib abrogates the CD4⁺ T cell-priming capacity of dendritic cells.
Spleen tyrosine kinase (SYK) is a core signalling protein that drives inflammatory responses and is fundamental to the propagation of signals via numerous immune receptors, including the B cell receptor and Fc receptors (FcRs). Fostamatinib, a small molecule SYK inhibitor, has shown evidence of ameliorating inflammation in RA patients. We sought to understand how the active metabolite of fostamatinib, R406, affects the inflammatory response at the cellular level. Antigen-specific in vivo systems and in vitro fluorescence microscopy were combined to investigate the effects of fostamatinib on antigen-specific interactions between dendritic cells (DCs) and CD4(+) T cells. Although it has previously been shown that R406 reduces the response of DCs to immune complexes (ICs), we found that fostamatinib failed to reduce specific CD4(+) T cell proliferation in mice after immunization with ICs. However, we observed in vitro that R406 reduces both the area and duration of cellular interactions between IC-activated DCs and specific CD4(+) T cells during the initial phase of cellular crosstalk. This led to diminished proliferation of antigen-specific CD4(+) T cells after R406 treatment compared with vehicle controls. This decreased proliferative capacity of CD4(+) T cells was accompanied by reduced expression of the co-stimulatory molecules, inducible T cell co-stimulator (ICOS) and PD-1, and abrogation of the production of inflammatory cytokines such as IFN-γ and IL-17. Our findings indicate a potential mechanism by which this compound may be effective in inhibiting FcR-driven CD4(+) T cell responses.


[Intraoperative diagnosis of incipient stomach cancer].
A combination procedure of surgical fibrogastroscopy was employed for diagnosis of incipient gastric carcinoma. It involved a routine examination of the mucosa, transillumination (examination in transmitted light) and endoscopy of the illuminated surgically-exposed stomach, with the endoscope's ligh being switched off. The study conducted in 24 patients with incipient carcinoma showed the effectiveness of the said diagnostic procedure in identifying foci of such lesions, their anatomic pattern as well as the extent of involvement of different layers of the stomach wall, submucosal and serous muscular layer.


Improved δ-valerolactam templates for the assembly of Aβ-miniamyloids by boronic ester formation.
The 6,7,8,8a-cis (all-cis) substituted δ-valerolactams of type 10, 11 and 12 are high-affinity diols for boronic ester formation, superior to the corresponding 6,7-trans analogues 1, 3 and 4. X-ray and NMR structure analysis have identified the differences of the six-membered ring conformations which cause the improved esterification properties of the all-cis stereoisomers. The homooligomeric all-cisδ-valerolactams 46-48 are used as polyol templates for the self-assembly of peptidic oligomers 49-52 by dynamic covalent chemistry. The templates have a diol spacing of approximately 5 Å, suitable for the assembly of branched peptides from the quantitative reaction between the peptide of interest, 2-formylphenylboronic acid and the respective template. According to this strategy, the tetrameric Aβ-miniamyloid 52 formed spontaneously from nine individual molecules in a three-component system. A detailed NMR analysis based on the complete sequential assignment of the trimeric Aβ(32-40)-miniamyloid 51 identified its three-dimensional structure in solution.


Chronic gestational exposure to ethanol impairs insulin-stimulated survival and mitochondrial function in cerebellar neurons.
Chronic gestational exposure to ethanol has profound adverse effects on brain development. In this regard, studies using in vitro models of ethanol exposure demonstrated impaired insulin signaling mechanisms associated with increased apoptosis and reduced mitochondrial function in neuronal cells. To determine the relevance of these findings to fetal alcohol syndrome, we examined mechanisms of insulin-stimulated neuronal survival and mitochondrial function using a rat model of chronic gestational exposure to ethanol. In ethanol-exposed pups, the cerebellar hemispheres were hypoplastic and exhibited increased apoptosis. Isolated cerebellar neurons were cultured to selectively evaluate insulin responsiveness. Gestational exposure to ethanol inhibited insulin-stimulated neuronal viability, mitochondrial function, Calcein AM retention (membrane integrity), and GAPDH expression, and increased dihydrorosamine fluorescence (oxidative stress) and pro-apoptosis gene expression (p53, Fas-receptor, and Fas-ligand). In addition, neuronal cultures generated from ethanol-exposed pups had reduced levels of insulin-stimulated Akt, GSK-3beta, and BAD phosphorylation, and increased levels of non-phosphorylated (activated) GSK-3beta and BAD protein expression. The aggregate results suggest that insulin-stimulated central nervous system neuronal survival mechanisms are significantly impaired by chronic gestational exposure to ethanol, and that the abnormalities in insulin signaling mechanisms persist in the early postnatal period, which is critical for brain development.


Identification of a plasma signature of psychotic disorder in children and adolescents from the Avon Longitudinal Study of Parents and Children (ALSPAC) cohort.
The identification of an early biomarker of psychotic disorder is important as early treatment is associated with improved patient outcome. Metabolomic and lipidomic approaches in combination with multivariate statistical analysis were applied to identify plasma alterations in children (age 11) (38 cases vs 67 controls) and adolescents (age 18) (36 cases vs 117 controls) preceeding or coincident with the development of psychotic disorder (PD) at age 18 in the Avon Longitudinal Study of Parents and Children (ALSPAC). Overall, 179 lipids were identified at age 11, with 32 found to be significantly altered between the control and PD groups. Following correction for multiple comparisons, 8 of these lipids remained significant (lysophosphatidlycholines (LPCs) LPC(18:1), LPC(18:2), LPC(20:3); phosphatidlycholines (PCs) PC(32:2; PC(34:2), PC(36:4), PC(0-34-3) and sphingomyelin (SM) SM(d18:1/24:0)), all of which were elevated in the PD group. At age 18, 23 lipids were significantly different between the control and PD groups, although none remained significant following correction for multiple comparisons. In conclusion, the findings indicate that the lipidome is altered in the blood during childhood, long before the development of psychotic disorder. LPCs in particular are elevated in those who develop PD, indicating inflammatory abnormalities and altered phospholipid metabolism. These findings were not found at age 18, suggesting there may be ongoing alterations in the pathophysiological processes from prodrome to onset of PD.


[The effect of UV-irradiation on telomerase activity and other stress-related proteins in human lens epithelial cells].
To investigate the changes and the role of telomerase activity and other stress-related proteins in the process of UV-induced DNA damage and repair in human lens epithelial cells. Human lens epithelial cells were irradiated at UV-doses 0.0 (control group) and 0.5, 1.5, 2.5, 3.5, 5.0, 7.5, 10.0 mJ/cm(2) (treated 1-7 group). Telomerase activity was determined by Telomerase Repeat Amplification Protocol-Enzyme Linked Immunosorbent Assay (TRAP-ELISA), p53, growth arrest and DNA damage inducible (GADD45), proliferating cell nuclear antigen (PCNA) and p16 protein levels were analyzed by Western blotting. Telomerase activity in control group and treated 1-7 group showed increased tendency, the differences of telomerase activity in 8 groups were significantly (P < 0.01). The expression of p53, GADD45, PCNA, p16 proteins showed increased tendency in experimental group, comparing with the control group, there were significant difference (P < 0.01). During UV-induced DNA damage and repair in human lens epithelial cells, telomerase activity was upregulated and the expression of stress-related proteins levels was increased. Upregulated telomerase activity may play both a protective and a proliferative role in human lens epithelial cells. Increased stress-related proteins level is critic in UV-induced DNA damage and repair in human lens epithelial. Increased telomerase activity is associated with increased levels of the stress-related proteins.


Effects of expressive writing intervention for women's PTSD, depression, anxiety and stress related to pregnancy: A meta-analysis of randomized controlled trials.
Expressive writing (EW) is a common psychological intervention that aims to improve the mental health of traumatized individuals. Pregnancy is considered an anxious or traumatic experience for some women, and any crisis in relation to pregnancy is closely associated with their psychological well-being. Post-traumatic stress disorder (PTSD), depression, anxiety and stress are the most prominent emotional and psychological responses that may occur during the process. However, the effects of EW in mediating women's PTSD, depression, anxiety and stress related to pregnancy remain uncertain, and no qualified meta-analysis has assessed such effects. The aim of this study was to assess the effectiveness of EW as a psychological intervention for women. Five databases, including PubMed, EMBASE, Cochrane, Web of Science and PsycINFO, were searched from inception to September 2019 for eligible studies. Finally, a total of 929 participants from 8 randomized controlled trial (RCT) studies were included. A pooled analysis demonstrated that EW was an efficient therapy for decreasing PTSD. However, the results showed that the EW intervention was not associated with the expected effects on anxiety and stress symptoms. The efficacy of EW for depression was inconclusive. More RCTs are necessary to verify the effectiveness of EW for depression. Studies concentrating on EW's effects on physical symptoms are necessary, and researchers should create an EW intervention group, neutral writing group and no writing group to examine the true effects of EW. Future research should examine whether longer, more frequent, and more targeted writing interventions would result in different conclusions.


Rehabilitation medicine: 3. Management of adult spasticity.
Spasticity refers to an abnormal, velocity-dependent (i.e., how fast the joint is moved through its range) increase in muscle tone resulting from interruption of the neural circuitry regulating the muscles and is a common complication of cerebral palsy, brain injuries, spinal cord injuries, multiple sclerosis and stroke. The muscle stretch reflex is thought to play an important role in spasticity generation. Spasticity can have a significant detrimental effect on daily functions, such as feeding, dressing, hygiene, bladder and bowel control, and mobility; patients' need for support can also influence the cost of care. Thus, managing these patients appropriately or referring them to those with expertise in this area is important. In this article, I review the pathophysiology of spasticity and the evaluation and management of adult patients with the condition. Two hypothetical cases are presented to illustrate the management of spasticity.


Effect of high-pressure processing and thermal treatment on quality attributes and nutritional compounds of "Songold" plum purée.
The application of hydrostatic high pressure on a "Songold" plum purée was assessed in comparison with heat pasteurization. To simulate industrial conditions, one-half of the total purée was manufactured with a pretreatment of thermal blanching (TB) and the other half without it (nonthermally blanched, NTB). Changes after thermal treatment and high-pressure processing (HPP: 400, 600 MPa) and after 20 d of refrigerated storage were evaluated. HPP maintained the microbial stability of the purées until the end of the storage period. Polyphenol oxidase activity was lower in TB purées than NTB purées. No treatment was completely effective to stop the enzyme activity, although a significant reduction was reached. Thermally treated purées showed more intense color changes after processing and storage than HP-treated purées. After processing, high-pressure (HP) purées treated at 600 MPa (TB and NTB) increased the extractability of carotenoids compared with initial untreated purée. Nevertheless, at the end of the storage, the highest carotenoid content was found in the TB purée treated at 400 MPa. After processing, total polyphenol levels were similar in all purées. TB and 600 MPa processing was more effective in the maintenance of the polyphenols than the other purées. TB increased the level of antioxidants after storage, compared to NTB purées. A previous TB step is necessary to inactivate browning enzymes before HPP to maintain the levels of bioactive compounds. HPP of plum purée could be a suitable alternative to traditional thermal processing, but more studies are necessary to ensure a major inactivation of polyphenol oxidase. High-pressure processing is one of the most successful technologies to obtain high-quality fruit purées without appreciable losses in taste, flavor, color, and nutritive value. However, for the introduction of a new technology, some advantages compared to the traditional thermal treatment need to be emphasized. The application of this technology could be reduced due to resistance of certain enzymes to pressurization. For this reason, in some cases the application of a previous thermal blanching can be necessary at industrial level. However, this can reduce the advantages of HPP application. This paper provides interesting information about the storage stability of plum purées after high-pressure treatments and assesses the need for applying heat pretreatments.


Joint haplotype assembly and genotype calling via sequential Monte Carlo algorithm.
Genetic variations predispose individuals to hereditary diseases, play important role in the development of complex diseases, and impact drug metabolism. The full information about the DNA variations in the genome of an individual is given by haplotypes, the ordered lists of single nucleotide polymorphisms (SNPs) located on chromosomes. Affordable high-throughput DNA sequencing technologies enable routine acquisition of data needed for the assembly of single individual haplotypes. However, state-of-the-art high-throughput sequencing platforms generate data that is erroneous, which induces uncertainty in the SNP and genotype calling procedures and, ultimately, adversely affect the accuracy of haplotyping. When inferring haplotype phase information, the vast majority of the existing techniques for haplotype assembly assume that the genotype information is correct. This motivates the development of methods capable of joint genotype calling and haplotype assembly. We present a haplotype assembly algorithm, ParticleHap, that relies on a probabilistic description of the sequencing data to jointly infer genotypes and assemble the most likely haplotypes. Our method employs a deterministic sequential Monte Carlo algorithm that associates single nucleotide polymorphisms with haplotypes by exhaustively exploring all possible extensions of the partial haplotypes. The algorithm relies on genotype likelihoods rather than on often erroneously called genotypes, thus ensuring a more accurate assembly of the haplotypes. Results on both the 1000 Genomes Project experimental data as well as simulation studies demonstrate that the proposed approach enables highly accurate solutions to the haplotype assembly problem while being computationally efficient and scalable, generally outperforming existing methods in terms of both accuracy and speed. The developed probabilistic framework and sequential Monte Carlo algorithm enable joint haplotype assembly and genotyping in a computationally efficient manner. Our results demonstrate fast and highly accurate haplotype assembly aided by the re-examination of erroneously called genotypes. A C code implementation of ParticleHap will be available for download from https://sites.google.com/site/asynoeun/particlehap.


Application of gonadotropin releasing hormone in hypogonadotropic hypogonadism--diagnostic and therapeutic aspects.
Puberty is the result of reactivation of the gonadotropin releasing hormone (GnRH) pulse generator resulting in an increasing release of GnRH by the hypothalamus, which stimulates the gonadotropic cells of the pituitary to synthesize and secrete LH and FSH. Hypogonadotropic hypogonadism (HH) is often the result of GnRH deficiency. The clinical picture is characterized by the absence of pubertal development and infertility. It is difficult to differentiate HH from delayed puberty since low gonadotropin and low testosterone levels are found in both conditions. We hypothesized that long-term GnRH administration may differentiate between the two conditions by a difference in the increase of gonadotropins, the idea being that in normal delayed puberty the pituitary of the patient has been primed with GnRH during the fetal and early postnatal period. Seventeen adolescents suspected of having hypogonadotropic hypogonadism were treated with pulsatile GnRH for 7 days. At the present time, the diagnosis of these patients is known and the results of the long-term GnRH stimulation have been evaluated according to the present diagnosis. The results show that the increase in gonadotropins following GnRH treatment is similar in both conditions. Therefore, at a prepubertal age a normal delayed puberty cannot be distinguished from hypogonadotropic hypogonadism using long-term GnRH stimulation. Long-term pulsatile GnRH treatment is a physiological therapy for the induction of puberty. Unlike testosterone it has the advantage of stimulation of testicular growth and fertility, as well as virilization, in males. We have treated 68 male patients with HH with pulsatile GnRH. The results show testicular growth and virilization in all the patients and spermatogenesis in 58 patients. Wearing a portable pump is cumbersome. However, the patients were very motivated and adapted very easily to this inconvenience. When spermatogenesis had developed, GnRH treatment was changed to human chorionic gonadotropin (hCG) administration 1-2 times per week intramuscularly or subcutaneously. During hCG therapy spermatogenesis was maintained or even improved. At least ten patients fathered children. Pulsatile GnRH cannot distinguish between a normal delayed puberty and a hypothalamic defect in still prepubertal patients. Pulsatile GnRH offers an appropriate way to initiate testicular growth including virilization and fertility in males with hypogonadotropic hypogonadism.


Quality of life among women living with HIV/AIDS in Yangon, Myanmar.
Women living with HIV/AIDS have many problems, especially in their quality of life. Although they receive ARTI they still experience stress and are discriminated against in their communities. Stigmatization, social support and other individual factors such as disease progression, economic status and gender role are determinants of quality of life among women living with HIV/AIDS. This cross sectional study was aimed to measure the quality of life among women living with HIV/AIDS in Yangon and to identify the factors that are able to predict it. 172 women living with HIV/AIDS, were included in the present study and were interviewed by using questionnaires. The WHOQOL-BREFform with 26 items was usedfor assessing the quality of life, then t-test and regression analysis was used for statistical analysis. Overall quality of life all domain scores were presented with transformed score (0 to 100 scale) for comparison, and the mean score was 79.7. Physical domain was the highest with a score of 58.19, social domain was 56.49, psychological domain was 48.54 and environmental domain was 46.84. A total of 64.5% had experienced a low level of perceived stigma, and only 0.6% experienced a high level of perceived stigma. Only 7% had good social support and 22.1% had low support. Social support, last CD4 count, family income per month, age, stigma and duration of treatment were determined as the significant predictors on quality of life after controlling for other factors. The factors could explain 22% of the variation in quality of life. In the absence of good social networks, peer group support, family involvement and support, reduction of stigma through multi-sectored approaches, women with HIV/AIDS will continue to suffer poor quality of life, less enjoyment and poor life satisfaction.


Photosynthetic acclimation to warming in tropical forest tree seedlings.
Tropical forests have a mitigating effect on man-made climate change by acting as a carbon sink. For that effect to continue, tropical trees will have to acclimate to rising temperatures, but it is currently unknown whether they have this capacity. We grew seedlings of three tropical tree species over a range of temperature regimes (TGrowth = 25, 30, 35 °C) and measured the temperature response of photosynthetic CO2 uptake. All species showed signs of acclimation: the temperature-response curves shifted, such that the temperature at which photosynthesis peaked (TOpt) increased with increasing TGrowth. However, although TOpt shifted, it did not reach TGrowth at high temperature, and this difference between TOpt and TGrowth increased with increasing TGrowth, indicating that plants were operating at supra-optimal temperatures for photosynthesis when grown at high temperatures. The high-temperature CO2 compensation point did not increase with TGrowth. Hence, temperature-response curves narrowed with increasing TGrowth. TOpt correlated with the ratio of the RuBP regeneration capacity over the RuBP carboxylation capacity, suggesting that at high TGrowth photosynthetic electron transport rate associated with RuBP regeneration had greater control over net photosynthesis. The results show that although photosynthesis of tropical trees can acclimate to moderate warming, carbon gain decreases with more severe warming.


Structurally ordered PtSn intermetallic nanoparticles supported on ATO for efficient methanol oxidation reaction.
The development of cost-effective methanol oxidation reaction (MOR) catalysts with a high activity and stability is highly desirable for direct methanol fuel cells. In this study, the structurally ordered PtSn intermetallic nanoparticles supported on Sb-doped SnO2 (ATO) have been successfully synthesized in ethylene glycol (EG) solution at 200 °C. Pt NPs were firstly formed on ATO, followed by the transformation from Pt into hexagonal PtSn on the surface of ATO. The obtained structurally ordered PtSn intermetallic NPs supported on ATO demonstrate significantly enhanced MOR activity and durability in comparison with commercial Pt/C. Our PtSn intermetallic NPs supported on ATO show a MOR activity 4.1 times higher than that of commercial Pt/C catalysts. Accelerated durability tests indicate that the commercial Pt/C catalysts lose about 50% of their initial current density after 500 cycles while the Pt/ATO-200-3 h catalyst loses only about 15% of its initial current density. Our PtSn intermetallic NPs supported on ATO are also found to have higher CO tolerance than commercial Pt/C. This work demonstrates an important strategy to rationally design high-performance structurally ordered Pt-based intermetallic NP catalysts for fuel cells and other applications.


Compositional differences in particle-associated and free-living microbial assemblages from an extreme deep-ocean environment.
Relatively little information is available for the composition of microbial communities present in hadal environments, the deepest marine locations. Here we present a description of the phylogenetic diversity of particle-associated (> 3 µm) and free-living (3-0.22 µm) microorganisms present in a pelagic trench environment. Small subunit ribosomal RNA gene sequences were recovered from members of the Bacteria, Archaea and Eukarya obtained from a depth of 6000 m in the Puerto Rico Trench (PRT). Species richness estimates for the bacterial particle-associated fraction were greater compared with the free-living fraction and demonstrated statistically significant compositional differences, while the archaeal fractions were not found to be significantly different. The particle-associated fraction contained more Rhodobacterales and unclassified Myxococcales along with Bacteroidetes, Planctomycetes and chloroplast sequences, whereas the free-living fraction contained more Caulobacterales, Xanthomonadales and Burkholderiales, along with Marine Group A and Gemmatimonadetes. The Eukarya contained a high abundance of Basidiomycota Fungi 18S rRNA genes, as well as representatives from the super-groups Rhizaria, Excavata and Chromalveolata. A diverse clade of diplonemid flagellates was also identified from the eukaryotic phylotypes recovered, which was distinct from previously identified deep-sea pelagic diplonemid groups. The significance of these results to considerations of deep-sea microbial life and particle colonization is discussed in comparison to the few other deep-ocean phylogenetic surveys available.


The landscape of somatic mutations in infant MLL-rearranged acute lymphoblastic leukemias.
Infant acute lymphoblastic leukemia (ALL) with MLL rearrangements (MLL-R) represents a distinct leukemia with a poor prognosis. To define its mutational landscape, we performed whole-genome, exome, RNA and targeted DNA sequencing on 65 infants (47 MLL-R and 18 non-MLL-R cases) and 20 older children (MLL-R cases) with leukemia. Our data show that infant MLL-R ALL has one of the lowest frequencies of somatic mutations of any sequenced cancer, with the predominant leukemic clone carrying a mean of 1.3 non-silent mutations. Despite this paucity of mutations, we detected activating mutations in kinase-PI3K-RAS signaling pathway components in 47% of cases. Surprisingly, these mutations were often subclonal and were frequently lost at relapse. In contrast to infant cases, MLL-R leukemia in older children had more somatic mutations (mean of 6.5 mutations/case versus 1.3 mutations/case, P = 7.15 × 10(-5)) and had frequent mutations (45%) in epigenetic regulators, a category of genes that, with the exception of MLL, was rarely mutated in infant MLL-R ALL.


Evidence for an antagonism between caffeine and adenosine in the human cardiovascular system.
A randomized, double-blind and placebo-controlled study was performed in 10 normotensive male subjects to analyze a possible antagonism between caffeine and adenosine with respect to their effects on the cardiovascular system in humans. Caffeine alone, 250 mg intravenously (i.v.), increased blood pressure by 9/12 mm Hg, and resulted in a fall of heart rate (HR) of 3 beats/min. Plasma epinephrine (E) rose by 114% after caffeine. Adenosine alone, in an increasing dose of 0.04-0.16 mg/kg/min, induced an increase in systolic blood pressure (SBP) (17 mm Hg), and HR (33 beats/min), a moderate fall in diastolic blood pressure (DBP) (-4 mm Hg), and no change of mean arterial pressure (MAP). At the highest adenosine infusion rate, forearm blood flow, skin temperature (ST), and transcutaneous oxygen tension were lowered, whereas plasma (nor)epinephrine was increased 227.2 and 215.9%, respectively. Adenosine infusion after caffeine induced comparable effects, but the fractional adenosine-induced changes of SBP, HR, plasma catecholamines, plasma renin activity (PRA), and aldosterone all were significantly reduced by previous administration of caffeine. Our results indicate an antagonism between caffeine and adenosine in humans, which may support the suggestion that some circulatory effects of caffeine are caused by an interaction with endogenous adenosine.


The possible effect of SCN1A and SCN2A genetic variants on carbamazepine response among Khyber Pakhtunkhwa epileptic patients, Pakistan.
SCN1A (3184 A>G) and SCN2A (56G>A) gene encodes α subunit of the neuronal voltage-gated sodium channel, which is a target for carbamazepine (CBZ). Recent studies have demonstrated that polymorphism of SCN1A (3184 A>G) and SCN2A (56G>A) was associated with use of CBZ. However, it has not been determined whether the polymorphism affects CBZ or other antiepileptic drug responsiveness. The aim of the study was to establish whether the SCN1A (3184 A>G) and SCN2A (56G>A) polymorphisms of the SCN1A and SCN2A genes affect responsiveness to CBZ. SCN1A (3184 A>G) and SCN2A (56G>A) gene polymorphisms were genotyped in 93 Khyber Pakhtunkhwa epileptic patients treated with CBZ. The association between CBZ responsiveness and the polymorphism was estimated by adjusting for clinical factors affecting the outcome of therapy. The number of seizure episodes was documented at baseline, and the therapy of each of the 93 patients was followed up. The plasma level of CBZ was determined using reverse-phase high-performance liquid chromatography. SCN1A and SCN2A genes were genotyped using RFLP. Data were analyzed using Graph Pad Prism 6. Mean age of the patients was 18.6±9.3 at the 3rd month and 18.7±9.5 at the 6th month. The baseline dose of CBZ was 468±19.8 mg/d and titrated at the rate of 48±1.4 and 4.0±0.2 mg/d. The difference in plasma level of CBZ was significant (P=0.004) between 3rd and 6th month among different genotypes of SCN1A gene in nonresponder and responder patients. At the 3rd month of the therapy, the poor responders were more likely (P=0.003 and P=0.01) to have variants (3184AG and 3184GG) of SCN1A gene. Similarly, poor responsders were more likely (P=0.0007 and P=0.001) to have variant genotypes (56GA, 56AA) of SCN2A gene at the 3rd month of the therapy. This study demonstrated a significant association between the SCN1A (3184 AG and GG) and SCN2A (56GA and AA) genotype with CBZ-nonresponsive epilepsy.


Experiences of stigma and discrimination among caregivers of persons with schizophrenia in China: a field survey.
In China, caregivers for family members with schizophrenia play an important role in treatment and recovery but may experience stigma and discrimination simply because of their family relationship. The object of this study was to measure the degrees and correlates of stigma and discrimination experiences among this group. Four hundred twenty-seven caregivers participated in this hospital-based and cross-sectional study in Ningbo and Guangzhou, China. Data were collected by trained interviewers using fixed questionnaires. Stigma and discrimination experiences were measured by the Modified Consumer Experiences of Stigma Questionnaire (MCESQ). Caregivers' social support was measured by the Social Support Rating Scale. Parametric analysis, nonparametric analysis and multivariate linear regression were used. The mean (SD) score of MCESQ was 2.44(0.45), 2.91(0.71) for stigma experiences and 1.97(0.37) for discrimination experiences on a five-point score ("1 = never" and "5 = very often"). Approximately 65% of caregivers reported that they tried to conceal their family members' illness, and 71% lacked the support of friends. The experience of stigma was significantly negatively associated with the perceived social support of caregivers (standard β = -0.2,p<0.001). Caregivers who were children of the patients experienced fewer stigmas than other (standard β = -0.18, p<0.001). Urban residence (standard β = -0.12, p<0.01) and patients did not complete primary school education (standard β = -0.13, p<0.01) were negatively related with stigmas. In addition, stigma and discrimination was more experienced in Zhejiang than in Guangdong (p<0.05). In conclusion, this study performed that caregivers of people with schizophrenia in China experienced general stigmas and rare discrimination and found the relations with social support, kinship, patient's educational level and regional differences. More interventions and supports should been given to caregivers who are lack of social support, who live in rural area and who are the patients' parents, spouses or siblings.


Different EDC/NHS activation mechanisms between PAA and PMAA brushes and the following amidation reactions.
Infrared spectroscopy was applied to investigate the well-known EDC/NHS (N-ethyl-N'-(3-(dimethylamino)propyl)carbodiimide/N-hydroxysuccinimide) activation details of poly(acrylic acid) (PAA) and poly(methacrylic acid) (PMAA) brushes grafted on porous silicon. Succinimidyl ester (NHS-ester) is generally believed to be the dominant intermediate product, conveniently used to immobilize biomolecules containing free primary amino groups via amide linkage. To our surprise, the infrared spectral details revealed that the EDC/NHS activation of PMAA generated anhydride (estimated at around 76% yield and 70% composition), but not NHS-ester (around 5% yield and 11% composition) under the well-documented reaction conditions, as the predominant intermediate product. In contrast, EDC/NHS activation of PAA still follows the general rule, i.e., the expected NHS-ester is the dominant intermediate product (around 45% yield and 57% composition), anhydride the side product (40% yield and 28% composition), under the optimum reaction conditions. The following amidation on PAA-based NHS-esters with a model amine-containing compound, L-leucine methyl ester, generated approximately 70% amides and 30% carboxylates. In contrast, amidation of PAA- or PMAA-based anhydrides with L-leucine methyl ester only produced less than 30% amides but more than 70% carboxylates. The above reaction yields and percentage compositions were estimated by fitting the carbonyl stretching region with 5 possible species, NHS-ester, anhydride, N-acylurea, unreacted acid, unhydrolyzed tert-butyl ester, and using the Beer-Lambert law. The different surface chemistry mechanisms will bring significant effects on the performance of surface chemistry-derived devices such as biochips, biosensors, and biomaterials.


Peak water limits to freshwater withdrawal and use.
Freshwater resources are fundamental for maintaining human health, agricultural production, economic activity as well as critical ecosystem functions. As populations and economies grow, new constraints on water resources are appearing, raising questions about limits to water availability. Such resource questions are not new. The specter of "peak oil"--a peaking and then decline in oil production--has long been predicted and debated. We present here a detailed assessment and definition of three concepts of "peak water": peak renewable water, peak nonrenewable water, and peak ecological water. These concepts can help hydrologists, water managers, policy makers, and the public understand and manage different water systems more effectively and sustainably. Peak renewable water applies where flow constraints limit total water availability over time. Peak nonrenewable water is observable in groundwater systems where production rates substantially exceed natural recharge rates and where overpumping or contamination leads to a peak of production followed by a decline, similar to more traditional peak-oil curves. Peak "ecological" water is defined as the point beyond which the total costs of ecological disruptions and damages exceed the total value provided by human use of that water. Despite uncertainties in quantifying many of these costs and benefits in consistent ways, more and more watersheds appear to have already passed the point of peak water. Applying these concepts can help shift the way freshwater resources are managed toward more productive, equitable, efficient, and sustainable use.


Variant Presentations of the Exstrophy-Epispadias Complex: A 40-Year Experience.
To investigate the diagnosis, surgical management, and outcomes in patients with variant EEC. Variant presentations of the exstrophy-epispadias complex (EEC) span a wide range of abnormalities. The rarity and diversity of EEC variants can lead to challenges in the diagnosis and subsequent management of this population. The authors reviewed an institutional database of 1336 EEC patients from 1975 to 2018 for variant presentations of EEC. Variant presentations included those with skin covered bladder exstrophy (BE), duplicate bladders, superior vesical fistula, and epispadias with major bladder prolapse. Surgical management and outcomes were assessed. In total, 44 EEC variants were identified. Nineteen (43%) presented with a skin-covered BE variant. Five patients presented with duplicate BE, while 6 presented with superior vesical fistula. Fourteen patients (32%) presented with epispadias with major bladder prolapse. Overall, 36 (82%) EEC variants underwent primary bladder closure, at a median of 135 days after birth (range 1-2010), with 21 (58%) undergoing pelvic osteotomy. Primary closures were successful in 89% of cases. Continence procedures were performed in 17 patients. This includes 5 patients who underwent bladder augmentation. However even without a continence procedure, continence with volitional voiding was found in 8 patients. The most common EEC variant is the skin-covered form of BE. In order to expedite appropriate management, accurate diagnosis upon initial presentation is crucial. Still, successful surgical reconstruction often results in continence that is similar to, or better than, nonvariant EEC presentations.


[Some features of nuclear genome transcription at early stages of liver regeneration. Effect of cytoplasmic factors on the RNA-polymerase activity of isolated nuclei].
Cytosol from the regenerating liver significantly activates the RNA-polymerase activity of isolated nuclei in normal liver. This activation occurs immediately after partial hepatectomy, reaches its maximum after the 1st post-operative hour and is practically eliminated after 1,5 hrs. The effect is thermolabile and is completely blocked by a preliminary administration of cycloheximide. Fractionation of cytosol on DEAE-cellulose reveals significant chromatographic differences of the activating factor as compared to that described in literature, which is present both in normal and regenerating liver. An important role of this factor in reprogramming of the genome from the hepatocytes at the preproliferation stage is postulated.


Clinical characteristics of eosinophilic asthma exacerbations.
Airway eosinophilia is associated with an increased risk of asthma exacerbations; however, the impact on the severity of exacerbations is largely unknown. We describe the sputum inflammatory phenotype during asthma exacerbation and correlate it with severity and treatment response. Patients presenting to hospital with an asthma exacerbation were recruited during a 12-month period and followed up after 4 weeks. Induced sputum was collected at both visits. Patients underwent spirometry, arterial blood gas analysis, fractional exhaled nitric oxide analysis, white blood cell counts and a screening for common respiratory viruses and bacteria. An eosinophilic exacerbation (EE) was defined as having sputum eosinophils ≥ 3% and a non-eosinophilic exacerbation as < 3% (NEE). A total of 47 patients were enrolled; 37 (79%) had successful sputum induction at baseline, of whom 43% had sputum eosinophils ≥3% (EE). Patients with EE had a significantly lower forced expiratory volume in 1 s (FEV1 ) % predicted (70.8%, P = 0.03) than patients with NEE (83.6%). Furthermore, EE patients were more likely to require supplemental oxygen during admission (63% vs 14%, P = 0.002). The prevalence of respiratory viruses was the same in EE and NEE patients (44% vs 52%, P = 0.60), as was bacterial infection (6% vs 14%, P = 0.44). Fractional expiratory nitric oxide (FeNO) correlated with sputum %-eosinophils (ρ = 0.57, P < 0.001), and predicted airway eosinophilia with a sensitivity of 86% and a specificity of 70%. Our findings suggest that eosinophilic asthma exacerbations may be clinically more severe than NEEs, supporting the identification of these higher risk patients for specific interventions.


DockStar: a novel ILP-based integrative method for structural modeling of multimolecular protein complexes.
Atomic resolution modeling of large multimolecular assemblies is a key task in Structural Cell Biology. Experimental techniques can provide atomic resolution structures of single proteins and small complexes, or low resolution data of large multimolecular complexes. We present a novel integrative computational modeling method, which integrates both low and high resolution experimental data. The algorithm accepts as input atomic resolution structures of the individual subunits obtained from X-ray, NMR or homology modeling, and interaction data between the subunits obtained from mass spectrometry. The optimal assembly of the individual subunits is formulated as an Integer Linear Programming task. The method was tested on several representative complexes, both in the bound and unbound cases. It placed correctly most of the subunits of multimolecular complexes of up to 16 subunits and significantly outperformed the CombDock and Haddock multimolecular docking methods. http://bioinfo3d.cs.tau.ac.il/DockStar email@example.com or firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Gas-chromatographic analysis for succinimide anticonvulsants in serum: macro- and micro-scale methods.
A gas-chromatographic analysis for the succinimide anticonvulsant drugs--ethosuximide, methsuximide, and phensuximide--in 1.0 ml of serum was modified to improve its reliability, speed, and precision. A separate procedure for 10-100 mul of serum was also developed. Neither method requires an initial preparation of derivatives. The working range of each method is about 10-100 mg/liter for the macro-method, 2-100 mg/liter for the micro-method. The methsuximide metabolite, N-desmethylmethsuximide, is included in both methods. Concentrations of N-desmethylmethsuximide in the blood of two patients with petit mal epilepsy are reported.


Optimized use of multi-functional positioning frame features for scoliosis surgeries.
A multi-functional positioning frame (MFPF) has been developed which includes a number of positioning features allowing for hip flexion and extension, thorax vertical displacement, lateral leg displacement, pelvic torsion and thorax lateral displacement. The objective of this study was to develop a method allowing for optimized combined use of the MFPF features. Finite element models (FEMs) representing the osseo-ligamentous structures of the spine, ribcage, pelvis and lower limbs, including muscles, were created for three different curve types (main thoracic, double major, and triple major) using a radiographic bi-planar reconstruction technique. Each FEM was subjected to an experimental design in which MFPF features were independently and simultaneously varied between extreme positions and the resultant changes in spinal geometry measured. Optimization of individual spinal geometrical parameters showed variability between curve types and some patterns such as minimum Cobb with lower limbs displaced laterally towards the convexity, pelvis raised on the side of concavity, and thorax laterally displaced towards the thoracic concavity. A weighted and normalized global optimization equation was developed which accounts for the relative importance and desired values of each geometrical parameter. Combined use of MFPF features and adjustments offers a wider range of possible intra-operative spinal geometries than their individual use.


Conflict and human African trypanosomiasis.
Human African Trypanosomiasis (HAT) has reemerged in sub-Saharan Africa as a disease of major public health importance. The success of HAT elimination in sub-Saharan Africa is subject to the feasibility of controlling, eliminating, or mitigating the determinants of incidence in affected countries. Conflict has been widely recognized and cited as a contributing factor to the resurgence of HAT in many countries, as well as to continuing HAT incidence in politically unstable and resource-poor regions. Despite extensive anecdotal and qualitative recognition of the role of conflict, there has been no quantitative research of this topic at the population level in affected African countries. We characterize the qualitative and quantitative associations between HAT incidence and conflict-related processes in HAT-affected African countries over the past 30 years. HAT and conflict-related data were collected for 35 affected countries in sub-Saharan Africa for the years 1976-2004. Descriptive and univariate inferential statistics, as well as negative binomial regression modeling, are used to assess the associations between HAT and conflict. A space-time scan statistic is used to identify significant incidence clusters. Clusters of HAT incidence over the past 30 years have predominantly coincided with periods of conflict or socio-political instability. HAT cases occurred significantly more often in countries and during years with conflict, high political terror, and internationalized civil war. The results indicate a lag period between the start of conflict events and a peak in incidence of approximately 10 years. We recommend explicit consideration and quantification of socio-political measures such as conflict and terror indices in GIS (Geographic Information Systems)-based risk assessments for HAT policy and intervention.


Analysis of Oligosaccharides in Korean Fermented Soybean Products by the Combination of Mass Spectrometry and Gas Chromatography.
Doenjang (fermented soybean paste) and ganjang (soy sauce) are traditional fermented foods that are widely consumed in Korea. The oligosaccharides found in soybean and its fermented foods have great potential to improve the quality of foods; however, their structural details have not been well studied. In this study, we used advanced mass spectrometry and gas chromatography to analyze oligosaccharides and their monomeric composition in two fermented soybean products. In both foods, oligosaccharides with a degree of polymerization ranging from 3 to 7 were found. Their constituent monosaccharides were characterized; galactose, xylose, arabinose, and rhamnose were the predominant constituents of the oligosaccharides, and fucose, fructose, mannose, glucose, and N-acetylglucosamine were also found. The great structural diversity of the oligosaccharides found suggests that soybean carbohydrates are hydrolyzed and/or transformed during fermentation and may yield novel oligosaccharides.


Pharmacokinetics of enrofloxacin following intravenous administration to greater rheas: a preliminary study.
The pharmacokinetic behaviour of enrofloxacin (ENR) and its active metabolite ciprofloxacin (CIP) were determined in six greater rheas following a single intravenous (i.v.) dose of 15 mg/kg bw. Plasma concentrations of ENR and CIP were simultaneously determined by a HPLC/u.v. method. Following i.v. administration, the plasma drug concentrations were best fitted by an open two-compartment model with a rapid distribution phase. The high volume of distribution (V(ss)=5.01 L/Kg) suggests good tissue penetration. ENR presents a high clearance (3.95 L/kg h) explaining the low AUC values (3.57 mg h/L) and a short permanence (t(1/2beta)=2.66 h and MRT=1.23 h). Ciprofloxacin comprised 14% of the total fluoroquinolone (ENR+CIP).


Galangin Inhibits LPS-Induced MMP-9 Expression via Suppressing Protein Kinase-Dependent AP-1 and FoxO1 Activation in Rat Brain Astrocytes.
Neuroinflammation, characterized by the increased expression of inflammatory proteins such as matrix metalloproteinases (MMPs), plays a critical role in neurodegenerative disorders. Lipopolysaccharide (LPS) has been shown to upregulate MMP-9 expression through the activation of various transcription factors, including activator protein 1 (AP-1) and forkhead box protein O1 (FoxO1). The flavonoid 3,5,7-trihydroxy-2-phenyl-4H-1-benzopyran-4-one (galangin) has been demonstrated to possess antioxidant and anti-inflammatory properties in various types of cells. Here, we investigated the mechanisms underlying the inhibitory effect of galangin on LPS-induced MMP-9 expression in rat brain astrocytes (RBA-1 cells). Pharmacological inhibitors and siRNAs were employed to explore the effects of galangin on LPS-challenged RBA-1 cells. Gelatin zymography, Western blotting, real-time PCR, and a luciferase reporter assay were used to detect MMP-9 activity, protein expression, mRNA levels, and promoter activity, respectively. The protein kinases involved in the LPS-induced MMP-9 expression were determined by Western blot. A chromatin immunoprecipitation (ChIP) assay was employed to evaluate the activity of c-Jun at the MMP-9 promoter. Galangin treatment attenuated the LPS-mediated induction of MMP-9 protein and mRNA expression, as well as the activity at the MMP-9 promoter. In addition, galangin exerted its inhibitory effects on MMP-9 expression through suppressing the LPS-stimulated activation of proline-rich tyrosine kinase (Pyk2), platelet-derived growth factor receptor beta (PDGFRβ), phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), mammalian target of rapamycin (mTOR), and mitogen-activated protein kinases (MAPKs). Pretreatment with galangin attenuated the LPS-induced phosphorylation of c-Jun and FoxO1. LPS-induced cell migration was also suppressed by galangin pretreatment. Galangin attenuates the LPS-induced inflammatory responses, including the induction of MMP-9 expression and cell migration, via inhibiting Pyk2/PDGFRβ/PI3K/Akt/mTOR/JNK1/JNK2 and p44/p42 MAPK cascade-dependent AP-1 and FoxO1 activities. These results provide new insights into the mechanisms through which galangin mitigates LPS-induced inflammatory responses, and suggest novel strategies for the management of LPS-related brain diseases.


Steady-state pharmacokinetics and tolerability of modafinil administered alone or in combination with dextroamphetamine in healthy volunteers.
The potential for a drug-drug interaction between modafinil and dextroamphetamine, each at steady state, was investigated in an open-label, randomized, single-period studyin 32 healthy male and female volunteers. All subjects received modafinil orally once daily for 28 days (200 mg on Days 1-7; 400 mg on Days 8-28). On Days 22 to 28, half of the subjects also received dextroamphetamine (20 mg) orally 7 hours after modafinil. Samples for pharmacokinetic (PK) profiling were obtained on Days 21 and 28. The mean changes in PK parameters for modafinil and its two circulating metabolites between the two groups were not statistically significantly different, except Cmax for modafinil acid. Adverse events obtained in the two groups were similar and mild or moderate in nature. The results indicate that administration of low-dose dextroamphetamine in this dosing regimen does not alter the steady-state pharmacokinetics of modafinil. The combination has a similar tolerability profile as modafinil alone.


Ontogenetic development, sexual differentiation, and effects of Aroclor 1254 exposure on expression of the arylhydrocarbon receptor and of the arylhydrocarbon receptor nuclear translocator in the rat hypothalamus.
Interaction of polychlorinated biphenyls (PCBs) with the aryl hydrocarbon receptor (AhR)/nuclear translocator (ARNT) system might interfere with the mechanisms controlling the sexual differentiation of the developing hypothalamus. The aim of this study was to evaluate the presence of AhR/ARNT in brain cells and the developmental profile of their expression in the hypothalamus of male and female rats during the perinatal period. Brain accumulation of the main PCB congeners after prenatal exposure to Aroclor 1254 and its influence on hypothalamic expression of AhR/ARNT was also assessed. The results show that: (a) AhR and ARNT are expressed both in neurons and in glia; (b) AhR expression progressively increases in the developing hypothalamus particularly in males, while ARNT is relatively constant in both sexes; (c) the prenatal administration of Aroclor to dams produces a differential accumulation of PCBs, depending on the chlorine atom number, and stimulates AhR expression only in the male hypothalamus. In conclusion, the developing male hypothalamus might be more sensitive to disrupting potential of PCBs.


Extrarenal pseudoaneurysm of the arterial anastomosis in a renal transplant: endovascular coil embolization with balloon remodeling technique.
Anastomotic pseudoaneurysms of transplanted kidneys are a very rare complication and encountered in less than 1% of such operations. They may be devastating and cause functional impairment and even loss of the graft. In this report, we present the first case of treatment of extrarenal pseudoaneurysm of arterial anastomosis in a renal transplant patient with endovascular coil embolization with the balloon remodeling technique. This method is mostly used in the treatment of wide-neck intracranial aneurysms.


Realization of efficient metal grating couplers for membrane-based integrated photonics.
Grating couplers are widely used to couple light between photonic integrated circuits and optical fibers. Here, we fabricate and characterize a device based on a buried metal grating. In contrast to dielectric gratings, simulations predict strongly reduced parasitic leakage of light to the substrate and are performance independent of the optical buffer thickness, while using standard fabrication processes with high yield. The gratings show a 3 dB bandwidth of 61 nm and chip-to-fiber coupling efficiency of 54%, which makes them attractive building blocks for on-wafer testing and dense optical interconnects.


A karyosystematic analysis of some water beetles related to Deronectes Sharp (Coleoptera, Dytiscidae).
An account is given of the karyotypes of five species and one additional subspecies of Deronectes Sharp, 1882, three species of Stictotarsus Zimmermann, 1919, one species of Trichonectes Guignot, 1941, four species of Scarodytes Gozis, 1914 and 17 species of Nebrioporus Régimbart, 1906. Deronectes species are characterised by a neo-Xy system of sex chromosomes and autosome numbers ranging from 60 (Deronectes ferrugineus Fery et Brancucci, 1987 and Deronectes wewalkai Fery et Fresneda, 1988) through 48 (Deronectes latus (Stephens, 1829), Deronectes angusi Fery et Brancucci, 1990) to 28 (Deronectes costipennis Brancucci, 1983, Deronectes costipennis gignouxi Fery et Brancucci, 1989 and Deronectes platynotus (Germar, 1834)). The three species of Stictotarsus, Stictotarsus duodecimpustulatus (Fabricius, 1792), Stictotarsus procerus (Aubé, 1838) and Stictotarsus bertrandi (Legros, 1956), all belonging to the Stictotarsus duodecimpustulatus group of species, have karyotypes comprising 54 autosomes and neo-Xy sex chromosomes. Trichonectes otini Guignot, 1941 has 48 autosomes and an X0 system of sex chromosomes, an arrangement shared with the 17 species of Nebrioporus Régimbart. The four Scarodytes species, Scarodytes halensis (Fabricius, 1787), Scarodytes nigriventris (Zimmermann, 1919), Scarodytes fuscitarsis (Aubé, 1838) and Scarodytes malickyi Wewalka, 1997, all have 54 autosomes and X0 sex chromosomes. The karyotypes of the various species are found to be distinctive and to support separation of these species from one another. In two cases (Nebrioporus martinii (Fairmaire, 1858) and Nebrioporus sardus (Gemminger et Harold, 1868), and Scarodytes halensis and Scarodytes fuscitarsis) the karyotypes require the recognition of the taxa as full species, not subspecies. The implications of these data for the generic classification are considered. The data are found to be compatible with the DNA-based phylogeny proposed by Ribera (2003), where the enlarged Stictotarsus proposed by Nilsson and Angus (1992) is found to be unsatisfactory.


High-resolution imaging of photoreceptors in macular microholes.
To assess photoreceptor structure in macular microholes by using adaptive optics scanning laser ophthalmoscopy (AO-SLO) and spectral-domain optical coherence tomography (SD-OCT) and compare with visual acuity. Fourteen eyes from 12 patients with macular microholes underwent a full ophthalmologic examination and imaging with a fundus camera, SD-OCT, and an original prototype AO-SLO system at each visit. All eyes had a cone outer segment tip line disruption and a normal retinal pigment epithelium line on SD-OCT images. Adaptive optics scanning laser ophthalmoscopy revealed foveal cone disruption (13 eyes, round or oval; 1 eye, T-shaped) in all eyes. Cone disruption area (mean = 14,805 ± 9120 μm(2); range, 3495-35,901 μm(2)) positively correlated with logMAR visual acuity at the first visit (P = 0.015, rs = 0.679). During the follow-up period, cone disruption area increased in two eyes, was stable in seven eyes, and decreased in five eyes. At the last visit, cone disruption area (mean = 8717 ± 7432 μm(2); range, 0-25,746 μm(2)) also positively correlated with logMAR visual acuity (P = 0.035, rs = 0.610). In one patient with bilateral microholes and no apparent vitreous traction, lesion size gradually increased. Cone disruption area decreased and visual acuity improved following oral prednisone therapy. Cone disruption occurs in eyes with macular microholes and a larger cone disruption area translates into a poorer visual acuity. Macular microholes, which are commonly observed as foveal cone inner and outer segment disruptions, may occur in eyes with or without vitreofoveal traction.


Chromatic second harmonic imaging.
We report a non-axial-scanning second harmonic imaging technique, in which the chromatic aberration of a Fresnel lens is exploited to focus different wavelengths of a fundamental beam into different axial positions to effectively realize axial scanning. Since the second harmonic signals at different axial positions are generated by different fundamental wavelengths and hence accordingly have different center wavelengths, they can be resolved and detected in parallel by using a spectrometer without axial mechanical scanning. We have demonstrated a system capable of achieving about 8 μm effective axial scanning range. Proof-of-concept imaging results are also presented.


Re-"Formate" T-cell Antitumor Responses.
Rowe and colleagues discover that one-carbon (1C) metabolism rewiring occurs upon T-cell activation to support proliferation and cytolytic activity in CD8+ T cells and that supplementation of 1C donor formate rescues the dysfunctional T cells and their responsiveness to anti-PD-1 in selective tumor-infiltrated T-cell subsets. This finding represents an attractive strategy to overcome a metabolic vulnerability in the tumor microenvironment and improve the efficacy of immune checkpoint blockade. See related article by Rowe et al., p. 2566 (8).


Simultaneous deletion of Bax and Bak is required to prevent apoptosis and interstitial fibrosis in obstructive nephropathy.
Proximal tubular injury and apoptosis are key mediators of the development of kidney fibrosis, a hallmark of chronic kidney disease. However, the molecular mechanism by which tubular apoptotic cell death leads to kidney fibrosis is poorly understood. In the present study, we tested the roles of Bcl-2-associated X (Bax) and Bcl-2 antagonist/killer (Bak), two crucial proteins involved in intrinsic apoptotic cell death, in the progression of kidney fibrosis. Mice with proximal tubule-specific Bax deletion, systemic deletion of Bak, and dual deletion of Bax and Bak were subjected to unilateral ureteral obstruction (UUO). Dual deficiency of Bax and Bak inhibited tubular apoptosis and atrophy. Consistent with decreased tubular injury, dual ablation of Bax and Bak suppressed UUO-induced inflammation and kidney fibrosis with decreased tubular cell cycle arrest, expression of fibrogenic and inflammatory cytokines, and oxidative stress in the kidney. Bax or Bak deficiency was insufficient to prevent apoptosis and all other aforementioned malevolent effects, suggesting compensatory mediation by each other in the respective signaling pathways. These data suggest that dual ablation of Bax and Bak in the kidney is required to prevent UUO-induced tubular apoptosis and the consequent kidney inflammation and fibrosis.


ESAT-6-dependent cytosolic pattern recognition drives noncognate tuberculosis control in vivo.
IFN-γ is a critical mediator of host defense against Mycobacterium tuberculosis (Mtb) infection. Antigen-specific CD4+ T cells have long been regarded as the main producer of IFN-γ in tuberculosis (TB), and CD4+ T cell immunity is the main target of current TB vaccine candidates. However, given the recent failures of such a TB vaccine candidate in clinical trials, strategies to harness CD4-independent mechanisms of protection should be included in future vaccine design. Here, we have reported that noncognate IFN-γ production by Mtb antigen-independent memory CD8+ T cells and NK cells is protective during Mtb infection and evaluated the mechanistic regulation of IFN-γ production by these cells in vivo. Transfer of arenavirus- or protein-specific CD8+ T cells or NK cells reduced the mortality and morbidity rates of mice highly susceptible to TB in an IFN-γ-dependent manner. Secretion of IFN-γ by these cell populations required IL-18, sensing of mycobacterial viability, Mtb protein 6-kDa early secretory antigenic target-mediated (ESAT-6-mediated) cytosolic contact, and activation of NLR family pyrin domain-containing protein 3 (NLRP3) inflammasomes in CD11c+ cell subsets. Neutralization of IL-18 abrogated protection in susceptible recipient mice that had received noncognate cells. Moreover, improved Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccine-induced protection was lost in the absence of ESAT-6-dependent cytosolic contact. Our findings provide a comprehensive mechanistic framework for antigen-independent IFN-γ secretion in response to Mtb with critical implications for future intervention strategies against TB.


Allergy to cereals and dairy products in adult, uncomplicated asthma: an epidemiological survey.
From a representative group from the adult population (34.958) of a town of 77.384 inhabitants, 99 persons were selected who: a) claimed to be periodically dyspneic independently with respect to respiratory tract infections. b) had normal chest x-rays and normal PE flow values. Approximately one year later, a history of atopic diseases was taken from 79 of them, and intradermal skin tests were performed with some inhalants and food allergens. At this time, 15 persons failed to report their previous complaints of dyspnea. Only 5 persons reported that various vegetables and pickles were responsible for their urticaria or eczema. Only 2 persons admitted various gastrointestinal symptoms, but non mentioned milk, eggs or cereals as possible causative agents of their bronchial, nasal, skin or circulatory symptoms. Distinctly positive immediate type skin reactions to the mixed milk and egg allergen were recorded in 19 patients (24%), and to cereals in 11 persons (14%). They all reacted strongly also to the house dust allergen, but they did not differ from all the remaining subjects in the intensity of their skin response to the control solution of histamine. People with positive skin reactions to cereals or dairy products complained of chronic or recurrent rhinitis much more often than the others (73% and 63% compared with 35%). Forty-five (47%) of them had a positive personal history of urticaria or eczema. These "food reactors" did not differ from "non reactors" in the frequency of the elevated serum IgE level which was raised in 66% of the whole group (of 79). The results suggest that allergy to cereals and dairy products may often be underestimated in adult asthmatics especially when positive reactions to inhalants are also present. The problems of reliability of the skin tests and history taking in food allergy are briefly discussed.


Molecular mechanisms for the p38-induced cellular senescence in normal human fibroblast.
We previously reported that TAK1, one of the mitogen-activated protein kinase kinase kinases (MAP3Ks), represses the transcription of the human telomerase reverse transcriptase (hTERT) gene in human cancer cells and induces cellular senescence in normal diploid human cells. On the basis of these results, we presumed a link between hTERT repression and the induction of cellular senescence. In this study, we identified the MAPK p38 as a downstream mediator of TAK1, which represses hTERT transcription. Further, we observed that hTERT expression was repressed in senescent normal human fibroblast, and was attenuated on treatment with SB203580, a p38-specific inhibitor, which suggests that p38 represses hTERT expression during cellular senescence. Next, we demonstrated that repression of hTERT, irrespective of the activation status of p38, is important for the induction of cellular senescence, by using hTERT-overexpressing cells and hTERT-knockdown cells. Our results suggested that p38 is activated during the serial passagings of normal human fibroblast, which results in the repression of hTERT transcription and induction of cellular senescence.


Escape from an Optoelectronic Tweezer Trap: experimental results and simulations.
Optoelectronic tweezers (OET) are a microsystem actuation technology capable of moving microparticles at mm s-1 velocities with nN forces. In this work, we analyze the behavior of particles manipulated by negative dielectrophoresis (DEP) forces in an OET trap. A user-friendly computer interface was developed to generate a circular rotating light pattern to control the movement of the particles, allowing their force profiles to be conveniently measured. Three-dimensional simulations were carried out to clarify the experimental results, and the DEP forces acting on the particles were simulated by integrating the Maxwell stress tensor. The simulations matched the experimental results and enabled the determination of a new "hopping" mechanism for particle-escape from the trap. As indicated by the simulations, there exists a vertical DEP force at the edge of the light pattern that pushes up particles to a region with a smaller horizontal DEP force. We propose that this phenomenon will be important to consider for the design of OET micromanipulation experiments for a wide range of applications.