Effect of letrozole on spermogram parameters and hormonal profile in infertile men: A clinical trial study.
There is no reliable treatment for men with idiopathic infertility, but the relationship between severe sperm production and the ratio of estrogen to testosterone levels has been shown. Aromatase is an enzyme that plays an important role in converting testosterone to estradiol and androstenedione to estrogen. Aromatase inhibitors can increase testosterone and androgen production without increasing the amount of estrogen in circulation. The aim of this study was to evaluate the effect of aromatase inhibitor letrozole on the male infertility. This pre- and post-quasi-experimental clinical trial was carried out on 41 men with an infertility diagnosis. The basic hormonal profile included FSH, LH, testosterone, and estradiol. The ratio of testosterone to serum estradiol was also calculated and recorded. The sperm analysis was performed before the treatment and the seminal parameters were evaluated and recorded. Patients were then treated with letrozole 2.5 mg daily for 4 months. At the end of 4th months, the hormonal profile was studied and seminal analysis performed and recorded. The levels of FSH, LH, testosterone, and estradiol, and the ratio of testosterone to estradiol increased significantly after letrozole treatment. The sperm concentration, sperm motility, and sperm forward motion significantly increased after letrozole treatment. Sperm morphology only lightly altered. The ratio of testosterone to estradiol levels in infertile men treated with aromatase inhibitor improved and caused changes in sperm parameters. Letrozole may be used to improve sperm parameters in infertile men with low serum testosterone to estradiol ratio.


Trace lithium is inversely associated with male suicide after adjustment of climatic factors.
Previously, we showed the inverse association between lithium in drinking water and male suicide in Kyushu Island. The narrow variation in meteorological factors of Kyushu Island and a considerable amount of evidence regarding the role of the factors on suicide provoked the necessities of adjusting the association by the wide variation in sunshine, temperature, rain fall, and snow fall. To keep the wide variation in meteorological factors, we combined the data of Kyushu (the southernmost city is Itoman, 26°) and Hokkaido (the northernmost city is Wakkanai, 45°). Multiple regression analyses were used to predict suicide SMRs (total, male and female) by lithium levels in drinking water and meteorological factors. After adjustment of meteorological factors, lithium levels were significantly and inversely associated with male suicide SMRs, but not with total or female suicide SMRs, across the 153 cities of Hokkaido and Kyushu Islands. Moreover, annual total sunshine and annual mean temperature were significantly and inversely associated with male suicide SMRs whereas annual total rainfall was significantly and directly associated with male suicide SMRs. The limitations of the present study include the lack of data relevant to lithium levels in food and the proportion of the population who drank tap water and their consumption habits. The present findings suggest that trace lithium is inversely associated with male but not female suicide after adjustment of meteorological factors.


Screening for cerebrovascular disease in microcephalic osteodysplastic primordial dwarfism type II (MOPD II): an evidence-based proposal.
Microcephalic osteodysplastic primordial dwarfism type II (OMIM 210720) is a rare autosomal recessive condition frequently associated with early-onset cerebrovascular disease. Presymptomatic detection and intervention could prevent the adverse consequences associated with this. We reviewed published cases of microcephalic osteodysplastic primordial dwarfism type II to ascertain prevalence and characteristics of cerebrovascular disease and use these data to propose an evidence-based approach to cerebrovascular screening. Of 147 cases identified, 47 had cerebrovascular disease (32%), including occlusive arteriopathy (including moyamoya) and cerebral aneurysmal disease. Occlusive disease occurred in younger individuals, and progression can be both rapid and clinically silent. A reasonable screening approach would be magnetic resonance imaging and angiography of the cervical and intracranial circulation at diagnosis, repeated at yearly intervals until 10 years, and every 2 years thereafter, unless clinical concerns occur earlier. At present it would appear that this needs to be life-long. Families and professionals should be alerted to the potential significance of neurologic symptoms and measures should be taken to maintain good vascular health in affected individuals.


Mechanistic and Structural Insights into the Prion-Disaggregase Activity of Hsp104.
Hsp104 is a dynamic ring translocase and hexameric AAA+ protein found in yeast, which couples ATP hydrolysis to disassembly and reactivation of proteins trapped in soluble preamyloid oligomers, disordered protein aggregates, and stable amyloid or prion conformers. Here, we highlight advances in our structural understanding of Hsp104 and how Hsp104 deconstructs Sup35 prions. Although the atomic structure of Hsp104 hexamers remains uncertain, volumetric reconstruction of Hsp104 hexamers in ATPγS, ADP-AlFx (ATP hydrolysis transition-state mimic), and ADP via small-angle x-ray scattering has revealed a peristaltic pumping motion upon ATP hydrolysis. This pumping motion likely drives directional substrate translocation across the central Hsp104 channel. Hsp104 initially engages Sup35 prions immediately C-terminal to their cross-β structure. Directional pulling by Hsp104 then resolves N-terminal cross-β structure in a stepwise manner. First, Hsp104 fragments the prion. Second, Hsp104 unfolds cross-β structure. Third, Hsp104 releases soluble Sup35. Deletion of the Hsp104 N-terminal domain yields a hypomorphic disaggregase, Hsp104(∆N), with an altered pumping mechanism. Hsp104(∆N) fragments Sup35 prions without unfolding cross-β structure or releasing soluble Sup35. Moreover, Hsp104(∆N) activity cannot be enhanced by mutations in the middle domain that potentiate disaggregase activity. Thus, the N-terminal domain is critical for the full repertoire of Hsp104 activities.


Relative merits of MRI, transrectal endosonography and CT in diagnosis and staging of carcinoma of prostate.
Magnetic resonance imaging (MRI) and transrectal sonography of 27 patients with biopsy-proved carcinoma of the prostate were performed to compare the sensitivity of these modalities to each other for diagnosis and to computed tomography (CT) for staging. Sonography was superior to MRI for the detection of intraglandular carcinoma and capsular disruption. MRI was superior to both sonography and CT for evaluating seminal vesicle invasion, and slightly better than CT for detecting lymphadenopathy.


Mycorrhizal fungal community composition in seven orchid species inhabiting Song Mountain, Beijing, China.
Mycorrhizal fungi play an important role in the germination and growth of orchids essentially influencing their survival, abundance, and spatial distribution. In this study, we investigated the composition of the mycorrhizal fungal community in seven terrestrial orchid species inhabiting Song Mountain, Beijing, China, using Illumina MiSeq high-throughput sequencing. The mycorrhizal communities in the seven orchids were mainly composed of members of the Ceratobasidiaceae, Sebacinales, and Tulasnellaceae, while a number of ectomycorrhizal fungi belonging to the Russulaceae, Tricholomataceae, Thelephoraceae, and Cortinariaceae were occasionally observed. However, the dominant fungal associates and mycorrhizal community differed significantly among the orchid species as well as subhabitats. These findings confirm the previous observation that sympatric orchid species show different preferences for mycorrhizal fungi, which may drive niche partitioning and contribute to their cooccurrence.


Between a Rock and Hard Place: Combined Effects of Authentic Leadership, Organizational Identification, and Team Prototypicality on Managerial Prohibitive Voice.
Managers are installed by the organization's stakeholders and shareholders to increase the organization's value; at the same time, they depend on their subordinates' acceptance to fulfill this leadership role. If the interest of the organization collides with the interest of their team, some managers act in the interest of their followers accepting potential disadvantages for their organizations and/or external stakeholders. In two experimental studies comprised mainly of German (N = 111) and US (N = 323) managers, we examined combined effects of authentic leadership, organizational identification, and self-perceived team prototypicality on managerial integrity operationalized as expressing work-related concerns to prevent organizations from harm (i.e., managerial voice). Our results show direct effects of authentic leadership and organizational identification on voice behavior across both studies. Furthermore, organizational identification increased voice for managers' low in authentic leadership pointing at a compensation effect. Finally, leader team prototypicality decreased the effect of identification on voice for managers high in authentic leadership but increased voice for managers low in authentic leadership, but only if these managers identified with their organization. In sum, our findings complement prior research that focused mainly on safety and instrumentality concerns by emphasizing the relevance of self-related antecedents of managerial voice.


Time course of expiratory propofol after bolus injection as measured by ion molecule reaction mass spectrometry.
Propofol in exhaled breath can be detected and monitored in real time by ion molecule reaction mass spectrometry (IMR-MS). In addition, propofol concentration in exhaled breath is tightly correlated with propofol concentration in plasma. Therefore, real-time monitoring of expiratory propofol could be useful for titrating intravenous anesthesia, but only if concentration changes in plasma can be determined in exhaled breath without significant delay. To evaluate the utility of IMR-MS during non-steady-state conditions, we measured the time course of both expiratory propofol concentration and the processed electroencephalography (EEG) as a surrogate outcome for propofol effect after an IV bolus induction of propofol. Twenty-one patients scheduled for routine surgery were observed after a bolus of 2.5 mg kg(-1) propofol for induction of anesthesia. Expiratory propofol was measured using IMR-MS and the cerebral propofol effect was estimated using the bispectral index (BIS). Primary endpoints were time to detection of expiratory propofol and time to onset of propofol's effect on BIS, and the secondary endpoint was time to peak effect (highest expiratory propofol or lowest BIS). Expiratory propofol and changes in BIS were first detected at 43 ± 21 and 49 ± 11 s after bolus injection, respectively (P = 0.29). Peak propofol concentrations (9.2 ± 2.4 parts-per-billion) and lowest BIS values (23 ± 4) were reached after 208 ± 57 and 219 ± 62 s, respectively (P = 0.57). Expiratory propofol concentrations measured by IMR-MS have similar times to detection and peak concentrations compared with propofol effect as measured by the processed EEG (BIS). This suggests that expiratory propofol concentrations may be useful for titrating intravenous anesthesia.


Platelet GPIb complex as a target for anti-thrombotic drug development.
Specific inhibition of platelet function is a major target of anti-thrombotic drug research. Platelet receptors are both accessible and specific but have multiple functions often linked to a wide range of ligands. GPIb complex is best known as a major platelet receptor for von Willebrand factor essential for platelet adhesion under high shear conditions found in arteries and in thrombosis. Recent animal studies have supported inhibition of GPIb as a good candidate for anti-thrombotic drug development with several classes of proteins showing important specific effects and the required discrimination between roles in haemostasis and thrombosis is important to protect against bleeding complications. These include antibodies, several classes of snake venom proteins, mutant thrombin molecules and peptides affecting subunit interactions. However, due to the nature of its receptor-ligand interactions involving large protein-protein interfaces, the possibility of developing classic pharmaceutical inhibitors for long term (and perhaps oral) treatment is still unclear, and additional information about structural interactions and signalling mechanisms is essential.


Acute and post-acute behavioral and psychological effects of salvinorin A in humans.
Salvia divinorum has been used for centuries, and nontraditional use in modern societies is increasing. Inebriation and aftereffects of use are poorly documented in the scientific literature. This double-blind, placebo-controlled, randomized study analyzed subjective experiences of salvinorin A (SA) inebriation and consequences of use after 8 weeks. Thirty middle-aged, well-educated, hallucinogen-experienced participants smoked either 1,017 or 100 μg SA 2 weeks apart in counterbalanced order. Vital signs were recorded before and after inhalation. A researcher rated participants' behavior during sessions. Participants completed the Hallucinogen Rating Scale (HRS) assessing inebriation immediately after each session. Differences were analyzed between groups as functions of dose and time. After 8 weeks, participants were interviewed to determine reported consequences and aftereffects. Participants talked, laughed, and moved more often on an active dose. All six HRS clusters were significantly elevated on an active dose indicating hallucinogenic experiences. No significant adverse events were observed or reported by participants. The present results indicate similarities as well as differences between the subjective effects of S. divinorum and other hallucinogens. As a selective kappa opioid receptor agonist, SA may be useful for expanding understanding of the psychopharmacology and psychology of hallucinogenic states beyond serotonergic mechanisms.


Interactions of pro-inflammatory and vasoactive mediators with nitric oxide in the regulation of rat vascular permeability during laparotomy.
Inhibition of constitutive nitric oxide (NO) synthases by administration of N(G)-nitro-L-arginine methyl ester (L-NAME) during abdominal laparotomy provokes extensive vascular leakage in the rat gastrointestinal tract, assessed by the extravasation of [125I]human serum albumin. In the present study, the role of vasoactive or neutrophil-derived pro-inflammatory mediators in this process has been investigated. Administration of the thromboxane synthase inhibitor, 1-benzyl-imidazole (BZI, 25-50 mg kg(-1), s.c.), the platelet-activating factor (PAF) receptor antagonist, 3-[4-(2-chlorophenyl)-9-methyl-6H-thienol-[3,2-f][1,2,4]-triazolo- [4, 3-a][1,4]-diazepine-2-yl]-1-(4-morpholynil)-1-propionate (WEB 2086; 0.5-1 mg kg(-1), s.c.), the 5-lipoxygenase synthase inhibitor, N-(4-benzyloxybenzyl)-acetohydroxamic acid (BW A137C; 4-20 mg kg(-1), s.c.) or the vasopressin pressor receptor antagonist ([Mca(1), Tyr(Me)(2),Arg(8)]vasopressin/Manning peptide; 0.01-0.2 microg kg(-1), s.c.) dose-dependently reduced the intestinal plasma leakage provoked by L-NAME (5 mg kg(-1), s.c.), following a 5-cm abdominal laparotomy in anaesthetised rats. These findings suggest that constitutive NO synthase effectively counteracts the damaging actions on microvascular integrity of mediators, including thromboxanes, PAF, leukotrienes and vasopressin, released during surgical intervention.


How to wrinkle a cell: Emerging mechanisms of microridge morphogenesis.
Microridges are laterally elongated actin-based protrusions arranged in striking maze-like patterns on the apical surfaces of mucosal epithelial cells. Recent studies have begun to reveal the molecular and mechanical factors that regulate microridge morphogenesis and allow them to adopt their unique properties. Microridges form from the coalescence of short actin-filled precursor protrusions called pegs. Microridge morphogenesis requires the establishment of apicobasal polarity, cortical myosin contraction, and Arp2/3 activity. Mature microridges contain branched actin networks, keratin filaments, and plakin cytolinkers that likely connect the two cytoskeletal elements. Once formed, microridges rearrange by fission and fusion to form increasingly regular patterns. Their highly organized arrangement provides an exciting system in which to study the interplay between molecular signaling and physical forces in the formation of subcellular patterns.


Where Do Arthroscopy Editors Stand on Systematic Reviews of the Literature?
Systematic reviews seem overly prevalent and often inconclusive. Yet, well-performed reviews provide powerful answers to clinical questions, whereas the results of a single clinical trial may not be reliably reproducible. Thus, in balance, we rest highly in favor of rigorously performed synthetic studies that stand at the top of the evidence-based medicine hierarchy.


[The histodynamic of alopecia areata in the dependence on the griseofulvin-induced epithelial proliferation (author's transl)].
The histodynamic of the alopecia areata--without or with the medicamentous induction of the hair growth respectively--was investigated comparatively. 1. Griseofulvin induced an epithelial proliferation in alopecia areata, which is noticeable in the surface epithelium as well as in the follicular epithelium. 2. Under the influence of griseofulvin a deeper penetration of the follicles into the fatty tissue--in connexion with a cutan-subcutaneous volume increase--results; at the same time the supra- and infraseboglandular follicular areas are lengthened. 3. The matter for discussion is that these epithelial changes originate from an increase of metabolic activity or blood supply respectively in the adjacent cutan- and subcutaneous tissue.


Antibiofilm activity of coconut (Cocos nucifera Linn.) husk fibre extract.
In this study, antibiofilm activity of coconut husk extract (CHE) was tested by various assays in the laboratory. The effects of CHE on extracellular polymeric substance (EPS) production, hydrophobicity and adhesion ability of Pseudomonas sp., Alteromonas sp. and Gallionella sp. and the antimicrobial activity of the extract against these bacteria were assessed. CHE was found to possess antibacterial activity against all the bacterial strains and affected the EPS production. The CHE affected the growth of the biofilm-forming bacteria in a culture medium. The hydrophobicity of the bacterial cells was also changed due to the CHE treatment. The active compound of the CHE was characterised by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and fourier transform infrared (FT-IR) analysis. HPLC spectrum showed a single peak and the FT-IR spectrum indicated the presence of an OH-group-containing compound in the extract. In conclusion the CHE could be used as a source for the isolation of antifouling compounds.


Characterization of cytoplasmic male sterility of rice with Lead Rice cytoplasm in comparison with that with Chinsurah Boro II cytoplasm.
Rice with LD-type cytoplasmic male sterility (CMS) possesses the cytoplasm of 'Lead Rice' and its fertility is recovered by a nuclear fertility restorer gene Rf1. Rf1 promotes processing of a CMS-associated mitochondrial RNA of atp6-orf79, which consists of atp6 and orf79, in BT-CMS with the cytoplasm of 'Chinsurah Boro II'. In this study, we found that LD-cytoplasm contained a sequence variant of orf79 downstream of atp6. Northern blot analysis showed that atp6-orf79 RNA of LD-cytoplasm was co-transcribed and was processed in the presence of Rf1 in the same manner as in BT-cytoplasm. Western blot analysis showed that the ORF79 peptide did not accumulate in an LD-CMS line, while ORF79 accumulated in a BT-CMS line and was diminished by Rf1. These results suggest that accumulation of ORF79 is not the cause of CMS in LD-cytoplasm and the mechanism of male-sterility induction/fertility restoration in LD-CMS is different from that in BT-CMS.


Effects of low exposure to traffic related air pollution on childhood asthma onset by age 10 years.
Although NO2, a major traffic related air pollutant, has been associated with onset of childhood asthma, young children may be more susceptible to traffic related air pollution exposure compared to other individuals. We linked data from National Longitudinal Survey of Children and Youths Cycle 1-5 (1994-2003) and the National Air Pollution Surveillance Program to determine the association between NO2 exposure and either early or late onset childhood asthma phenotypes. Children diagnosed with asthma from age 0-3 were defined as having early onset asthma. Children diagnosed with asthma from age 4-9 were defined as having late onset asthma. Mean NO2 exposure for each quartile was 6.31 ppb, 9.45 ppb, 11.83 ppb, and 17.9 ppb. Higher levels of NO2 exposure were more strongly associated with early childhood asthma (Quartile 3 OR: 2.11, 95% CI: 1.29, 3.44, Quartile 4 OR: 2.16, 95% CI: 1.27, 3.68) compared to the lowest level of NO2 exposure (Quartile 1). No such association was observed with risk of late childhood asthma onset. Asthma susceptibility to NO2 exposure may vary with the childhood developmental stage, and young children may be susceptible to NO2 exposure at levels well below national and international guidelines. Our study emphasizes the importance of considering the timing of childhood asthma onset in future studies and confirms the increased risk of early onset of childhood asthma associated even with relatively low NO2 exposure levels.


Collagen promotes anti-PD-1/PD-L1 resistance in cancer through LAIR1-dependent CD8+ T cell exhaustion.
Tumor extracellular matrix has been associated with drug resistance and immune suppression. Here, proteomic and RNA profiling reveal increased collagen levels in lung tumors resistant to PD-1/PD-L1 blockade. Additionally, elevated collagen correlates with decreased total CD8+ T cells and increased exhausted CD8+ T cell subpopulations in murine and human lung tumors. Collagen-induced T cell exhaustion occurs through the receptor LAIR1, which is upregulated following CD18 interaction with collagen, and induces T cell exhaustion through SHP-1. Reduction in tumor collagen deposition through LOXL2 suppression increases T cell infiltration, diminishes exhausted T cells, and abrogates resistance to anti-PD-L1. Abrogating LAIR1 immunosuppression through LAIR2 overexpression or SHP-1 inhibition sensitizes resistant lung tumors to anti-PD-1. Clinically, increased collagen, LAIR1, and TIM-3 expression in melanoma patients treated with PD-1 blockade predict poorer survival and response. Our study identifies collagen and LAIR1 as potential markers for immunotherapy resistance and validates multiple promising therapeutic combinations.


Mutanase from Trichoderma harzianum inductively Produced by Mutan: Short-Term Treatment to Degrade Mature Streptococcus mutans Biofilm.
This study aimed to evaluate the disruptive effect of fungal mutanase against cariogenic biofilm after short-term treatment. For that, mature Streptococcus mutans biofilms (n = 9) were exposed to active or inactivated enzymes produced by Trichoderma harzianum for 1 min, two times per day. Biofilms were analyzed by amount of matrix water-insoluble polysaccharides, bacterial viability, acidogenicity, and morphology by scanning electron microscopy (SEM). The group treated with active enzymes (AE) had a significantly lower amount of insoluble polysaccharides (893.30 ± 293.69) when compared to the negative control group (NaCl, 2192.59 ± 361.96), yet no significant difference was found when comparing to the positive control group (CHX, 436.82 ± 151.07). Also, there was no significant effect on bacteria metabolism and viability (P-value < 0.05). Data generated by the quantitative analysis were confirmed through scanning electron microscopy images. Thus, fungal mutanase degraded the biofilm after a short-term treatment without interfering with bacterial viability and metabolism. Such findings offer insight to the development of routine oral care products containing this input.


Neural stem cell delivery via porous collagen scaffolds promotes neuronal differentiation and locomotion recovery in spinal cord injury.
Neural stem cell (NSC) grafts have demonstrated significant effects in animal models of spinal cord injury (SCI), yet their clinical translation remains challenging. Significant evidence suggests that the supporting matrix of NSC grafts has a crucial role in regulating NSC effects. Here we demonstrate that grafts based on porous collagen-based scaffolds (PCSs), similar to biomaterials utilized clinically in induced regeneration, can deliver and protect embryonic NSCs at SCI sites, leading to significant improvement in locomotion recovery in an experimental mouse SCI model, so that 12 weeks post-injury locomotion performance of implanted animals does not statistically differ from that of uninjured control animals. NSC-seeded PCS grafts can modulate key processes required to induce regeneration in SCI lesions including enhancing NSC neuronal differentiation and functional integration in vivo, enabling robust axonal elongation, and reducing astrogliosis. Our findings suggest that the efficacy and translational potential of emerging NSC-based SCI therapies could be enhanced by delivering NSC via scaffolds derived from well-characterized clinically proven PCS.


Bevacizumab: in first-line treatment of advanced and/or metastatic renal cell carcinoma.
Bevacizumab, an anti-vascular endothelial growth factor recombinant humanized monoclonal antibody, directly inhibits tumor angiogenesis and hence tumor growth. First-line therapy with intravenous bevacizumab 10 mg/kg every 2 weeks plus subcutaneous interferon-alpha-2a 9 million international units three times weekly has been evaluated in two randomized, double-blind or open-label, multicenter phase III trials (AVOREN [n = 649] and CALGB 90206 [n = 732]). Bevacizumab combination therapy resulted in a median progression-free survival that was significantly (p < or = 0.0001) longer than that seen with placebo plus interferon-[alpha]-2a in AVOREN (10.2 vs 5.4 months) [hazard ratio (HR) 0.63 (95% CI 0.52, 0.75)] and that seen with interferon-alpha-2a alone in CALGB 90206 (8.5 vs 5.2 months). Overall survival data in AVOREN and CALGB 90206 are not yet mature. In the interim overall survival analysis in AVOREN, median overall survival was 19.8 months with placebo plus interferon-alpha-2a, but had not yet been reached with bevacizumab plus interferon-alpha-2a (HR 0.79 [95% CI 0.62, 1.02; p = 0.0670]). The overall tumor response rate with bevacizumab plus interferon-alpha-2a was significantly (p < or = 0.0001) higher than with placebo plus interferon-alpha-2a in AVOREN (31% vs 13%) and that with interferon-alpha-2a alone in CALGB 90206 (25.5% vs 13.1%). Subgroup analyses in AVOREN suggested that interferon-alpha-2a dose reductions (to manage grade > or =3 adverse events attributable to the drug) did not compromise the efficacy of combination treatment with bevacizumab plus interferon-alpha-2a. The addition of bevacizumab to interferon-alpha-2a in AVOREN was generally well tolerated. No unexpected/new adverse events were observed; bevacizumab-associated toxicities were generally of mild intensity.


An intact helical domain is required for Gα14 to stimulate phospholipase Cβ.
Stimulation of phospholipase Cβ (PLCβ) by the activated α-subunit of Gq (Gαq) constitutes a major signaling pathway for cellular regulation, and structural studies have recently revealed the molecular interactions between PLCβ and Gαq. Yet, most of the PLCβ-interacting residues identified on Gαq are not unique to members of the Gαq family. Molecular modeling predicts that the core PLCβ-interacting residues located on the switch regions of Gαq are similarly positioned in Gαz which does not stimulate PLCβ. Using wild-type and constitutively active chimeras constructed between Gαz and Gα14, a member of the Gαq family, we examined if the PLCβ-interacting residues identified in Gαq are indeed essential. Four chimeras with the core PLCβ-interacting residues composed of Gαz sequences were capable of binding PLCβ2 and stimulating the formation of inositol trisphosphate. Surprisingly, all chimeras with a Gαz N-terminal half failed to functionally associate with PLCβ2, despite the fact that many of them contained the core PLCβ-interacting residues from Gα14. Further analyses revealed that the non-PLCβ2 interacting chimeras were capable of interacting with other effector molecules such as adenylyl cyclase and tetratricopeptide repeat 1, indicating that they could adopt a GTP-bound active conformation. Collectively, our study suggests that the previously identified PLCβ-interacting residues are insufficient to ensure productive interaction of Gα14 with PLCβ, while an intact N-terminal half of Gα14 is apparently required for PLCβ interaction.


Samarium-153-ethylene diamine tetramethylene phosphonate, a beta-emitting bone-targeted radiopharmaceutical, useful for patients with osteoblastic bone metastases.
Bone metastases are prevalent among cancer patients and frequently cause significant morbidity. Oncology providers must mitigate complications associated with bone metastases while limiting therapy-related adverse effects and their impact on quality of life. Multiple treatment modalities, including chemotherapy, surgery, external beam radiation therapy, and radioisotopes, among others, have been recommended and utilized for palliative treatment of bone metastases. Radioisotopes such as samarium-153 are commonly used in the setting of multifocal bone metastases due to their systemic distribution, affinity for osteoblastic lesions, acceptable toxicity profile, and convenience of administration. This review focuses on samarium-153, first defining its radiobiologic and pharmacokinetic properties before describing many clinical trials that support its use as a safe and effective tool in the palliation of patients with bone metastases.


Human big-endothelin-1 and endothelin-1 release prostacyclin via the activation of ET1 receptors in the rat perfused lung.
Although ET1 and ET2 binding sites were found in rat lung membranes, a selective ET1 receptor antagonist, BQ-123 (10 microM), did not displace [125I]-endothelin-1 ([125I]ET-1) from ET2 sites, illustrating the selectivity of the angatonist for ET1 receptors. In rat perfused lungs, BQ-123 (1 microM) markedly reduced the prostacyclin (PGI2) releasing properties of endothelin-1 (ET-1: 5 nM) and human big-ET-1 (100 nM) suggesting that both peptides induce the release of PGI2 via the selective activation of ET1 receptors.


The acute effect of a mineralocorticoid receptor agonist on corticotrope secretion in Addison's disease.
Mineralocorticoid receptors (MR) in the hippocampus display an important role in the control of hypothalamic-pituitary-adrenal (HPA) axis, mediating the ''proactive'' feedback of glucocorticoids (GC). Fludrocortisone (FC), a potent MR agonist, has been shown to decrease HPA activity through a hippocampal mechanism. Since it has been demonstrated that FC shows a significant inhibition of the HPA axis response to hCRH stimulus in normal subjects, also at doses usually administered as replacement therapy in patients with Addison's disease, an FC effect at MRs in human pituitary or a GR-pituitary agonism stronger than believed until now has been postulated. Ten patients affected by autoimmune Addison's disease received: (1) placebo p.o. + placebo i.v., (2) hydrocortisone (H) 10 mg p.o. + placebo i.v., (3) FC 0.1 mg p.o. + placebo i.v., (4) FC 0.1 mg and H 10 mg p.o. + placebo i.v. to verify a possible GR FC-mediated effect that might display a repercussion on the GC-replacement therapy. H reduced ACTH (p < 0.01) and increased cortisol levels (p < 0.01) with respect to the placebo session, while FC did not affect either ACTH or cortisol levels compared to placebo, and higher ACTH and lower cortisol levels (p < 0.03 and p < 0.01) were observed compared with the H session; furthermore the co-administration of FC + H showed ACTH and cortisol profiles similar to that observed during H alone. Our study showed a lack of FC effect on corticotrope secretion in Addison's disease, thus making unlikely the hypothesis of its GR pituitary agonism and the risk of glucocorticoid excess in primary adrenal insufficiency.


Venous thrombotic events and impact on outcomes in patients treated with first-line single-agent pembrolizumab in PD-L1 ≥ 50% advanced non small cell lung cancer.
Few data are available on the impact of venous thrombotic events (VTE) in patients with metastatic non-small cell lung cancer (mNSCLC) treated with immunotherapy. This is a secondary analysis of the ESKEYP study, a national, retrospective, multicenter study that consecutively included all PD-L1 ≥ 50% mNSCLC patients who initiated first-line treatment with pembrolizumab monotherapy. From May 2017 to November 2019, 845 patients were included (from availability of pembrolizumab in this indication in France to the authorization of the combination with chemotherapy). Impact of VTE and patient characteristics were analyzed. Of the 748 patients (88.5%) with available data, the incidence of VTE was 14.8% (111/748). At pembrolizumab initiation, Khorana score was ≥ 2 for 55.0% (61/111) of them. Recurrence of VTE was reported for 4 of the 111 patients and 5 had bleeding complications. Patients with VTE were significantly younger, had more frequently long-term corticosteroids treatment and more often liver metastases. Progression-free survival (PFS) was significantly shorter in patients with VTE compared to patients without VTE: 6.1 (95% CI 4.1-9.0) months vs. 8.3 (6.9-10.3) months (p = 0.03). VTE did not significantly impact overall survival (OS): 15.2 (10.0-24.7) months with VTE and 22.6 (18.4-29.8) months without VTE (p = 0.07). In multivariate analysis for PFS and OS, HRs for VTE were 1.3 (0.99-1.71), p = 0.06 and 1.32 (0.99-1.76), p = 0.05. The incidence of VTE appears to be as high with in first-line immunotherapy as with chemotherapy in patients with mNSCLC, with in patient with VTE, a no significant trend for lower PFS and OS in multivariate analysis. more marked impact on PFS than on OS.


Phase I, dose-escalating study of elisidepsin (Irvalec(®)), a plasma membrane-disrupting marine antitumor agent, in combination with erlotinib in patients with advanced malignant solid tumors.
To determine the recommended dose for phase II trials of elisidepsin (PM02734, Irvalec®) in combination with erlotinib in patients with advanced malignant solid tumors. Open-label, dose-escalating, phase I study of intravenous elisidepsin administered weekly (days 1, 8 and 15) over 3 h as a flat dose (FD) and daily oral erlotinib, every 3 weeks. A pharmacokinetic analysis was done on blood samples collected around the first elisidepsin infusion. Thirty patients were treated across six different dose levels (DLs) ranging from elisidepsin 0.33-2.25 mg/erlotinib 100-150 mg. Two patients had dose-limiting toxicities: grade 3 bilirubin increase (DL3: 0.75 mg/150 mg) and a dose omission for > 2 weeks due to grade 3 alanine aminotransferase increase (DL6: 2.25 mg/100 mg). The daily erlotinib dose was escalated to 150 mg at DL2-DL5, but decreased to 100 mg at DL6, as most grade 3 toxicities were related to this agent only. The most frequent toxicities were transaminase increases (related to elisidepsin), and rash, pruritus and diarrhea (related to erlotinib). No objective responses were observed. Despite no overlapping toxicities, the combination was declared unfeasible due to frequent elisidepsin dose delays. The pharmacokinetics of elisidepsin/erlotinib was not significantly different from that of each agent alone. The difficulty in combining elisidepsin with the standard dose of erlotinib (150 mg), together with the lack of antitumor activity, made the combination unattractive for further development. The trial was closed without having determined a recommended dose.


Signaling mechanisms of glucose-induced F-actin remodeling in pancreatic islet β cells.
The maintenance of whole-body glucose homeostasis is critical for survival, and is controlled by the coordination of multiple organs and endocrine systems. Pancreatic islet β cells secrete insulin in response to nutrient stimuli, and insulin then travels through the circulation promoting glucose uptake into insulin-responsive tissues such as liver, skeletal muscle and adipose. Many of the genes identified in human genome-wide association studies of diabetic individuals are directly associated with β cell survival and function, giving credence to the idea that β-cell dysfunction is central to the development of type 2 diabetes. As such, investigations into the mechanisms by which β cells sense glucose and secrete insulin in a regulated manner are a major focus of current diabetes research. In particular, recent discoveries of the detailed role and requirements for reorganization/remodeling of filamentous actin (F-actin) in the regulation of insulin release from the β cell have appeared at the forefront of islet function research, having lapsed in prior years due to technical limitations. Recent advances in live-cell imaging and specialized reagents have revealed localized F-actin remodeling to be a requisite for the normal biphasic pattern of nutrient-stimulated insulin secretion. This review will provide an historical look at the emergent focus on the role of the actin cytoskeleton and its regulation of insulin secretion, leading up to the cutting-edge research in progress in the field today.


Palmar dislocation of the scaphoid and lunate.
A case of palmar dislocation of the scaphoid and lunate was complicated by a delay in the diagnosis and avascular necrosis of the lunate. Anatomic reduction was obtained 34 days after the injury. At 2 1/2 years follow-up the patient had good return of hand function and radiologic evidence of resolution of the avascular necrosis without evidence of collapse or intercarpal arthritis.


Assessment of tumor invasion of the vena caval wall in renal cell carcinoma cases by magnetic resonance imaging.
We evaluated the role of magnetic resonance imaging (MRI) in patients with renal cancer and inferior vena caval involvement with reference to its ability to characterize the extent and nature of inferior vena caval tumor extension and wall invasion. The study included 12 consecutive patients with renal cancer and inferior vena caval involvement. All patients underwent imaging on a 1.5 Tesla MRI unit. Coronal, axial T1 and axial T2-weighted images were performed in all cases, while in 6 3-dimensional gadolinium enhanced magnetic resonance angiography and venography were also performed. Images were assessed for the extent and nature of tumor extension, that is tumor versus thrombus, and invasion of the inferior vena caval wall. Imaging results were compared with operative findings. On MRI the extent and nature of the inferior vena caval tumor was correctly defined in all cases. The sensitivity, specificity and accuracy of inferior vena caval wall invasion were 100%, 89% and 92%, respectively. In patients with renal cancer and inferior vena caval involvement MRI defines the tumor level in the inferior vena cava. It is also a sensitive technique for detecting vessel wall invasion and provides important preoperative information for surgical planning.


The evaluation and application of a radioimmunoassay for the measurement of diphenoxylic acid, the major metabolite of diphenoxylate hydrochloride (Lomotil), in human plasma.
Antibodies to diphenoxylic acid, the pharmacologically active metabolite of Lomotil, were successfully used to develop a precise and specific radioimmunoassay for the measurement of diphenoxylic acid in human plasma. The observed cross-reaction of the antiserum with Lomotil (23.5%) and p-hydroxy diphenoxylic acid (2.9%) was not considered to affect significantly the accuracy of the direct determination of diphenoxylic acid in plasma from human volunteers after ingestion of Lomotil tablets. Within-day and between-day coefficients of variation were better than 3 and 6%, respectively, over the concentration range of 3.4 to 255 ng ml-1. Comparable precision could be achieved at 2 ng ml-1 by doubling the volume of sample analyzed. The assay was used to measure plasma concentrations of diphenoxylic acid in 12 human volunteers for up to 24 hr after ingestion of Lomotil (10mg) tablets. Plasma diphenoxyllic acid levels rose to a mean (SE) maximum level of 87.8 (2.7) ng ml-1 3.3 (0.3) hr after dosing. By 24 hr after dosing plasma levls had decreased to 14.26 (1.67) ng ml-1. The appearance and elimination of plasma diphenoxylic acid could be described by a biexponential function. The appearance half-life was calculated to be 0.82 (0.09) hr, and the elimination half-life was 7.24 (0.73) hr.


Human mast cells express stem cell factor.
Stem cell factor (SCF) is a major cytokine regulator of mast cell growth and function. The present study demonstrates that human mast cells are able to produce SCF. Constitutive synthesis of SCF mRNA was seen in the mast cells isolated from human lung and skin by RT-PCR. This was confirmed by in situ hybridization in conjunctival mast cells of both tryptase-only (MCT) and tryptase/chymase (MCTC) subsets. SCF protein product was found in conjunctival MCT and MCTC mast cells by immunohistochemistry. Soluble SCF protein was detected in the culture supernatant of isolated lung mast cells by ELISA, and cross-linkage of IgE receptor (Fc epsilon-RI) on the lung mast cells in culture did not alter SCF mRNA expression, or the secreted soluble SCF protein. This was consistent with the finding that levels of SCF mRNA expression in conjunctival mast cells were similar between normal subjects and patients with seasonal allergic conjunctivitis (SAC). This study shows that human mast cells themselves are a cellular source of SCF, as well as being target cells for this growth factor. SCF may regulate mast cell growth and function via both paracrine and autocrine mechanisms. The production of SCF by mast cells may be regulated via mechanisms other than IgE receptor-mediated pathways.


Antiarrhythmic drug therapy of atrial fibrillation.
In the post-AFFIRM era, treatment of AF has become the treatment of symptoms. In some patients, this will be simple rate control, but there remain a significant cohort of patients in whom rate control alone does not give acceptable symptom relief. In this group, antiarrhythmic therapy still has a role, and the AFFIRM trial indicates that this therapeutic strategy is without significant deleterious effect on mortality. The choice of antiarrhythmic agent must be individualized according to underlying cardiac pathologies and comorbidities, however. Most recently, the introduction of dofetilide has widened the therapeutic options in patients with severe heart disease, and the Canadian Trial of Atrial Fibrillation indicated the superior efficacy of amiodarone at low doses. The release/ development of newer Class III antiarrhythmic agents may offer hope for the benefits of amiodarone without the serious adverse effects with long-term therapy.


Continuous colorimetric screening assays for the detection of specific L- or D-α-amino acid transaminases in enzyme libraries.
In the course of a project devoted to the stereoselective synthesis of non-proteinogenic α-amino acids using α-transaminases (α-TA), we report the design and optimization of generic high-throughput continuous assays for the screening of α-TA libraries. These assays are based on the use of L- or D-cysteine sulfinic acid (CSA) as irreversible amino donor and subsequent sulfite titration by colorimetry. The assays' quality was assessed under screening conditions. Hit selection thresholds were accurately determined for every couple of substrates and a library of 232 putative transaminases expressed in Escherichia coli host cells was screened. The reported high throughput screening assays proved very sensitive allowing the detection with high confidence of activities as low as 10 μU (i.e., 0.01 nmol substrate converted per min). The assays were also evidenced to be stereochemically discriminant since L-CSA and D-CSA allowed the exclusive detection of L-TA and D-TA, respectively. These generic assays thus allow testing the stereoselective conversion of a wide range of α-keto acids into α-amino acids of interest. As a proof of principle, the use of 2-oxo-4-phenylbutyric acid as acceptor substrate led to the identification of 54 new α-TA offering an access to valuable L- or D-homophenylalanine.


Anthracycline cardiotoxicity is exacerbated by global p38β genetic ablation in a sexually dimorphic manner but unaltered by cardiomyocyte-specific p38α loss.
Severe cardiotoxic effects limit the efficacy of doxorubicin (DOX) as a chemotherapeutic agent. Activation of intracellular stress signaling networks, including p38 mitogen-activated protein kinase (MAPK), has been implicated in DOX-induced cardiotoxicity (DIC). However, the roles of the individual p38 isoforms in DIC remain incompletely elucidated. We recently reported that global p38δ deletion protected female but not male mice from DIC, whereas global p38γ deletion did not significantly modulate it. Here we studied the in vivo roles of p38α and p38β in acute DIC. Male and female mice with cardiomyocyte-specific deletion of p38α or global deletion of p38β and their wild-type counterparts were injected with DOX. Survival and health were tracked for 10 days postinjection. Cardiac function was assessed by echocardiography and electrocardiography and fibrosis by Picrosirius red staining. Expression and activation of signaling proteins and inflammatory markers were measured by Western blot, phosphorylation array, and chemokine/cytokine array. Global p38β deletion significantly aggravated DIC and worsened cardiac electrical and mechanical function deterioration in female mice. Mechanistically, DIC in p38β-null female mice correlated with increased autophagy, sustained hyperactivation of proapoptotic JNK signaling, as well as remodeling of a myocardial inflammatory environment. In contrast, cardiomyocyte-specific deletion of p38α improved survival of DOX30-treated male mice 5 days posttreatment but did not influence cardiac function in DOX-treated male or female mice. Our data highlight the sex- and isoform-specific roles of p38α and p38β MAPKs in DOX-induced cardiac injury and suggest a novel in vivo function of p38β in protecting female mice from DIC.NEW & NOTEWORTHY We show that p38α and p38β have distinct in vivo functions in a murine model of acute DIC. Specifically, although conditional cardiomyocyte-specific p38α deletion exhibited mild cardioprotective effects in male mice, p38β deletion exacerbated the DOX cardiotoxicity in female mice. Our findings caution against employing pyridinyl imidazole inhibitors that target both p38α and p38β isoforms as a cardioprotective strategy against DIC. Such an approach could have undesirable sex-dependent effects, including attenuating p38β-dependent cardioprotection in females.


Measurement of tear resistance after manual capsulorhexis and femtosecond laser-assisted capsulotomy of crystalline lenses.
In this study, the tear resistance of porcine lens capsules after continuous curvilinear capsulorhexis (CCC) and femtosecond (fs)-laser-assisted capsulotomy for cataract surgery (FLC) with different laser parameters is measured with a custom-made testing setup. Forty-five fresh porcine lenses were randomly chosen for CCC (n = 15) or FLC 1 (n = 15) and FLC 2 (n = 15). The FLC 1-group was treated with smaller spot distances than the FLC 2-group. The force necessary to break the opening of the anterior capsule and the maximum displacement were measured. The mean tear resistance of the CCC-group (150 ± 70 mN) was higher than that of the FLC 1-group (60 ± 20 mN) and the FLC 2-group (30 ± 20 mN). It could be shown that CCC leads to a significantly higher tear resistance of the opening than FLC in porcine lenses. The femtosecond laser group demonstrated that smaller spot distances lead to a higher tear resistance.


Achyranthes aspera elevates thyroid hormone levels and decreases hepatic lipid peroxidation in male rats.
A study was made to evaluate the role of Achyranthes aspera on the changes in serum thyroid hormone concentrations and glucose levels in male rats. An attempt was also made to establish the relationship between hepatic lipid peroxidation and extract induced changes in thyroid hormone concentration, if any. Adult male Wistar rats were orally administered with the aqueous leaf extract of Achyranthes aspera at a dose of 200 mg/kg b. wt./day for 7 days. The effects of the extract on body weight, hepatic protein content, lipid peroxidation (LPO), superoxide dismutase (SOD) and catalase (CAT) activities and on serum triiodothyronine (T3), thyroxine (T4) and glucose levels were evaluated. The extract exhibited significant prothyroidic activity as it enhanced the levels of both the thyroid hormones along with an increase in serum glucose concentration, body weight and hepatic protein content. On the other hand, it decreased hepatic LPO without altering the activities of the two antioxidant enzymes, SOD and CAT significantly, suggesting a direct free radical scavenging activity of the extract. It appears that the Achyranthes aspera leaf extract is both prothyroidic and antiperoxidative in nature.


Wilson disease, ABCC2 c.3972C > T polymorphism and primary liver cancers: suggestions from a familial cluster.
Polymorphisms in genes modulating xenobiotics metabolism, in particular the ABCC2 c.3972C > T single nucleotide polymorphism (SNP) at exon 28, have been suggested to increase primary liver cancer (PLC) risk. Conversely, the occurrence of PLCs in Wilson disease patients is a rare event, in contrast with the occurrence observed in other chronic liver diseases. Here we report the clinical case of five siblings carrying the ABCC2 c.3972C > T SNP; three of them were affected by Wilson disease and two brothers with Wilson disease also developed PLCs. The presence of the ABCC2 c.3972C > T SNP was assessed by Sanger sequencing and the exposure of PLC risk factors by standardized questionnaires. Notably, PLCs occurred only in the two brothers with the ABCC2 c.3972C > T SNP and Wilson disease who resulted exposed to asbestos and cigarette smoking, but not in the other siblings with the ABCC2 c.3972C > T SNP, alone or in association with Wilson disease, not exposed to these carcinogens and/or to other known risk factors for PLCs. These findings suggest that ABCC2 c.3972C > T SNP and WD, also in association, may not represent a sufficient condition for PLC development, but that co-occurrence of further host/exogenous risk factors are needed to drive this process, reinforcing the notion that liver carcinogenesis is the result of a complex interplay between environmental and host genetic determinants. Due to the sporadic cases of this study and the paucity of data currently available in literature on this issue, future investigations in a larger population are needed to confirm our findings.


[Quantitative analysis of superposition of environmental task requirements].
Requirements for 1486 tasks have been registered with an Ergonomic Job Description Questionnaire (EJDQ). Frequencies of high and extremely high scaled environmental task requirements are analysed, related to branches, tasks and sex. The frequencies of superpositions are analysed for dominating requirements. Priorities for ergonomic research may be defined with the method presented.


Increased levels of serum clusterin is associated with intrauterine growth restriction and adverse pregnancy outcomes in preeclampsia.
Our aim was to investigate serum clusterin levels in preeclampsia and to determine whether any changes in clusterin levels are useful in distinguishing the presence of concomitant intrauterine growth restriction (IUGR) and in predicting adverse pregnancy outcomes. A prospective case-control study was carried out which included 86 pregnant women (47 patients with preeclampsia and 39 healthy controls). Maternal serum samples obtained from all participants and clusterin levels were determined by enzyme-linked immunosorbent assay (ELISA). Compared with controls, women with preeclampsia had significantly higher clusterin levels (mean 83.8±23.6 vs. 119.2±40.5, P<0.01). Further analysis revealed the highest clusterin levels were in patients with preeclampsia and IUGR (P<0.001). According to the receiver operating characteristic (ROC) analysis performed for the predictive value of clusterin levels for adverse maternal outcomes, the area under the curve (AUC) was 0.738 (95% CI: 0.616-0.859). The best clusterin cut-off value in predicting adverse maternal outcomes was 102.6 pg/mL with 75% sensitivity and 66% specifity. Multivariable logistic regression analysis revealed serum clusterin levels of >102.6 pg/mL was independently associated with preeclampsia (OR: 6.18, 95% CI: 2.41-15.9) and maternal adverse outcomes (OR: 5.13, 95% CI: 2.01-13.1) and also clusterin levels higher than 117.4 pg/mL were associated with adverse neonatal outcomes (OR: 5.02, 95% CI: 1.04-24.3). The current study suggests that increased levels of clusterin is associated with IUGR and probably predictive for adverse pregnancy outcomes in preeclampsia.


Diagnostic notes on the spider orb-weaving genera Carepalxis and Ocrepeira (Araneae: Araneidae), with description of three new species from Central Brazil.
Three new species of the araneid spider genera Carepalxis L. Koch, 1872 and Ocrepeira Marx, 1883 from Central Brazil are described and illustrated: Carepalxis topazio new species, based on a single female from Cristalina; C. quasimodo new species, from Braslia and Jaborandi based on females and a male, this being the first male of Carepalxis described in the American continent; and Ocrepeira planalto new species, based on males and females from Cavalcante, Cristalina, Jaborandi and Alto Paraso. A key to Carepalxis females in the New World is provided. Furthermore, the similarity between American Carepalxis and Ocrepeira is discussed, with diagnostic considerations.


Actin lessons from pathogens.
Salmonella typhimurium secretes proteins that co-opt the host actin cytoskeleton to induce membrane ruffling, leading to the uptake of the bacterium. New information about the biochemical activities of the Salmonella protein SipA suggests that this protein might inhibit host cell actin dynamics by competing with ADF/cofilin and gelsolin, two key proteins that promote the turnover of actin filaments.


Intracellular Platelet-Activating Factor Acetylhydrolase, Type II: A Unique Cellular Phospholipase A2 That Hydrolyzes Oxidatively Modified Phospholipids.
Intracellular platelet-activating factor acetylhydrolase, type II [PAF-AH (II)] is a monomeric 40kDa enzyme that was originally purified as an enzyme that hydrolyzes the acetyl group of PAF (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine). It can also hydrolyze phospholipids with short length or oxidatively modified sn-2 acyl chains, whereas it can hardly hydrolyze phospholipids with two long fatty acyl chains. PAF-AH (II) is the only phospholipase A2 that is myristoylated at its N-terminus and is evolutionarily conserved from lower organisms to mammals. Studies using cultured cells and mice suggest that PAF-AH (II) functions as an antioxidant phospholipase. Here, we will review our current understanding of PAF-AH (II) and discuss its potential functions in physiological and pathological conditions.


Modulation of Akt/mTOR signaling overcomes sunitinib resistance in renal and prostate cancer cells.
Tyrosine kinase inhibitors exhibit impressive activity against advanced renal cell carcinoma. However, recent clinical studies have shown an equivocal response to sunitinib in patients with castration-resistant prostate cancer. The tumor suppressor PTEN acts as a gatekeeper of the phosphoinositide 3-kinase (PI3K)/Akt/mTOR cell-survival pathway. Our experiments showed that PTEN expression inversely correlates with sunitinib resistance in renal and prostate cancer cells. Restoration of PTEN expression markedly increases sensitivity of tumor cells to sunitinib both in vitro and in vivo. In addition, pharmacologic manipulation of PI3K/Akt/mTOR signaling with PI3K/mTOR inhibitor, GDC-0980, mTOR inhibitor, temsirolimus, or pan-Akt inhibitor, GSK690693, was able to overcome sunitinib resistance in cancer cells. Our findings underscore the importance of PTEN expression in relation to sunitinib resistance and imply a direct cytotoxic effect by sunitinib on tumor cells in addition to its antiangiogenic actions.


Projections from the lateral geniculate nucleus to the hypothalamus of the Mongolian gerbil (Meriones unguiculatus): an anterograde and retrograde tracing study.
The lateral geniculate nucleus of the thalamus sends efferents to the hypothalamic suprachiasmatic nucleus, which is involved in generation and entrainment of several circadian rhythms. It seems reasonable to believe that the lateral geniculate conveys visual information about the length of the photoperiod to the circadian oscillator. In order to study in more detail the topographical relationship between the lateral geniculate and the suprachiasmatic nucleus, anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) and retrograde tracing with wheatgerm agglutinin coupled to horseradish peroxidase (WGA-HRP) were performed in the gerbil. After iontophoretic injections of PHA-L in the lateral geniculate, a large number of PHA-L-immunoreactive fibers and nerve terminals were observed in the ventrolateral part of the suprachiasmatic nucleus. Nerve fibers were also present in the ventromedial and dorsolateral portions, particularly in the caudal half of the nucleus. PHA-L-immunoreactive nerve fibers continued outside the borders of the suprachiasmatic nucleus to the adjacent anterior hypothalamic, the periventricular, and the subparaventricular areas. A moderate number of fibers entered the lateral hypothalamic area and the tuber cinerum via the optic tract and chiasm. Moreover, the paraventricular nucleus, the supraoptic nucleus, the medial preoptic area, the lateral preoptic area, and the supramammillary nucleus contained a few labeled fibers. In all parts of the hypothalamus receiving an input from the lateral geniculate, fine beaded immunoreactive fibers with varicosities and nerve terminals were observed, some of which were found in close apposition to hypothalamic neurons. Only after labeling of neurons in the intergeniculate leaflet of the lateral geniculate nucleus, fibers were found in the hypothalamus. This topographical organization of the geniculohypothalamic pathway was supported by retrograde tracing after injections of WGA-HRP in the suprachiasmatic area. In these experiments, retrograde labeled neurons were observed in the intergeniculate leaflet and, in agreement with the anterograde studies, most of labeling was observed in the ipsilateral side. These results confirm that the suprachiasmatic nucleus receives a substantial input from the intergeniculate leaflet of the lateral geniculate. Moreover, the present data demonstrate that the suprachiasmatic nucleus is not the only nucleus that receives a direct visual input. Thus other hypothalamic areas might be influenced by a direct rhythmic neuronal input as well.


Impact of several antibiotics and 2-bromoethanesulfonate on the volatile fatty acid degradation, methanogenesis and community structure during thermophilic anaerobic digestion.
The main aim of the present study was to gain insight into the stability of an anaerobic digestion process suffering from exposure to antibiotics and the methanogenic inhibitor 2-bromoethanesulfonate (BES). For this purpose, eleven antibiotics and BES were investigated with regard to the degradation of volatile fatty acids (VFAs), methanogenesis, and impact on the microbial community structure. Only neomycin, gentamicin, rifampicin, and BES showed complete inhibitions of VFA degradations. This points to distinct interferences with important trophic degradation cascades. Based upon DGGE and sequencing approaches, Methanosarcina spp. were severely influenced by the treatments while hydrogenotrophic methanogens were less affected. Interestingly, BES and neomycin inhibited the degradation of acetate while only BES inhibited methanogenesis completely. It seems that Methanosarcina spp. were mandatory for the degradation of acetate at high rates. The present results highly emphasize the detrimental effects of antimicrobial compounds with the potential to significantly inhibit the anaerobic digestion.


Role of midbrain raphe in stress-induced renin and prolactin secretion.
Stress-induced changes in renin and prolactin secretion were studied using a conditioned emotional response paradigm. Three minutes after being placed in a chamber, the stressed animals received a brief electric shock (1.0 mA for 10 s through the grid floor), then were returned to their home cage. This procedure was repeated for 3 consecutive days. On the fourth day, the rats were placed in the chamber for 3 min, but instead of receiving shock, they were removed and sacrificed. Control animals were treated in the same manner, except that they never received foot shock. The sham-operated stressed rats evidenced significant elevations in plasma renin activity (270%) and prolactin level (550%). Electrolytic lesions in the dorsal raphe nucleus blocked the stress-induced increase in plasma renin activity but did not affect the stress-induced increase in prolactin secretion. Electrolytic lesions in the median raphe nucleus did not affect prolactin levels in either control or stressed animals. However, median raphe lesions led to a significant increase in plasma renin activity in non-stressed rats and potentiated the stress-induced elevation in plasma renin activity. These results suggest that neurons within the dorsal and median raphe nuclei are involved in the regulation of renin but not prolactin secretion during stress. The results also suggest that median raphe neurons play a role in basal renin secretion.


Development and Elucidation of a Pd-Based Cyclization-Oxygenation Sequence for Natural Product Synthesis.
Pd-catalyzed sequences involving oxidative addition, cyclization, and termination through intermolecular nucleophile capture have tremendous utility. Indeed, they can generate a plethora of different polycyclic structures possessing a diverse range of functionality. However, one area of deficiency for Pd0 /PdII variants is the ability to conclude them with oxygen-based species. Inspired by the recent discovery of one such reaction in the course of a total synthesis program, we delineate herein that it has significant strength, both in terms of substrate scope as well as the terminating oxygen nucleophile. As a result, the reaction proved critical in achieving total syntheses of two oxygenated natural products, one of which was prone to over-oxidation. Finally, a mechanistic proposal that accounts for its success is provided.


Domain structure analysis of elongation factor-3 from Saccharomyces cerevisiae by limited proteolysis and differential scanning calorimetry.
Elongation-factor-3 (EF-3) is an essential factor of the fungal protein synthesis machinery. In this communication the structure of EF-3 from Saccharomyces cerevisiae is characterized by differential scanning calorimetry (DSC), ultracentrifugation, and limited tryptic digestion. DSC shows a major transition at a relatively low temperature of 39 degrees C, and a minor transition at 58 degrees C. Ultracentrifugation shows that EF-3 is a monomer; thus, these transitions could not reflect the unfolding or dissociation of a multimeric structure. EF-3 forms small aggregates, however, when incubated at room temperature for an extended period of time. Limited proteolysis of EF-3 with trypsin produced the first cleavage at the N-side of Gln775, generating a 90-kDa N-terminal fragment and a 33-kDa C-terminal fragment. The N-terminal fragment slowly undergoes further digestion generating two major bands, one at approximately 75 kDa and the other at approximately 55 kDa. The latter was unusually resistant to further tryptic digestion. The 33-kDa C-terminal fragment was highly sensitive to tryptic digestion. A 30-min tryptic digest showed that the N-terminal 60% of EF-3 was relatively inaccessible to trypsin, whereas the C-terminal 40% was readily digested. These results suggest a tight structure of the N-terminus, which may give rise to the 58 degrees C transition, and a loose structure of the C-terminus, giving rise to the 39 degrees C transition. Three potentially functional domains of the protein were relatively resistant to proteolysis: the supposed S5-homologous domain (Lys102-Ile368), the N-terminal ATP-binding cassette (Gly463-Lys622), and the aminoacyl-tRNA-synthase homologous domain (Glu820-Gly865). Both the basal and ribosome-stimulated ATPase activities were inactivated by trypsin, but the ribosome-stimulated activity was inactivated faster.


Cardiovascular effects of morphine and opioid peptides following intracisternal administration in chloralose-anesthetized rats.
Beta-Endorphin (0.9--2.0 nmol), morphine (11--250 nmol) and D-ala2-met-enkephalinamide (17--33 nmol) administered intracisternally produced preferential vasodepressor responses and bradycardia. Leu- (1.8--180 nmol), met-enkephalin (17--520 nmol) and alpha-endorphin (5.7--57 nmol) administered in the same way produced preferential vasopressor effects and the latter two peptides also produced bradycardia. Results obtained with naloxone (300 nmol) given intracisternally indicate that the pressor and depressor actions as well as the bradycardia are mediated through opiate receptors. The results indicate that opioid peptides may be involved in central cardiovascular control.


Right Information, Right Patient, Right Time: Utilizing the MyCareCompass Platform to Deliver Patient Education in the Oncology Setting.
Patient education is an important part of cancer care as it increases patient activation and informed decision-making, reduces anxiety, and improves outcomes. However, challenges to providing appropriate education to patients exist on both the health provider and patient side of the equation, e.g., time constraints and health literacy issues. Digital health education is a fast-growing field of interest that has been shown to improve health outcomes, increase effectiveness of medical treatments and education, lower medical costs, and enhance both clinical diagnosis and research opportunities by streamlining data collection, sharing, and analysis. In 2019, Fox Chase Cancer Center was selected by ARCHES, an established patient education software company, to pilot its award-winning digital patient engagement system MyCareCompass. During the pilot, patients scheduled for port insertions were sent electronic messages inviting them to review various online educational materials related to their procedure and subsequent concerns. The invitations and resources were seamlessly integrated into the scheduling system and timed to arrive when patients would most need them. There was high usage of the port-related materials and patients reported a high level of satisfaction with the delivery system and the information. This automated process of delivering high-quality and relevant patient education was able to be implemented smoothly with IT involvement, had a positive impact on patients without adding any extra burden to the care team, and highlighted opportunities to integrate these types of interventions into routine care.


Loss of pons-to-hypothalamic white matter tracks in brainstem obesity.
Hyperphagia and obesity have been reported following damage to the hypothalamus in humans. Other brain sites are also postulated to be involved in the control of food intake and body weight regulation, such as the amygdala and brainstem. The brainstem, however, is thought to primarily integrate short-term meal-related signals but not affect long-term alterations in body weight, which is controlled by higher centers. The objective of this study was to identify structural pathways damaged in a patient with a brainstem cavernoma who experienced sudden onset of hyperphagia and >50 kg weight gain in <1 year following surgical drainage via a midline suboccipital craniotomy. Diffusion tensor imaging revealed loss of nerve fiber connections between her brainstem, hypothalamus and higher brain centers with preservation of motor tracks. Imaging and endocrine testing confirmed normal hypothalamic structure and function. Gastric bypass surgery restored normal appetite and body weight to baseline. This is the first report of 'brainstem obesity' and adds to the brain regions that can determine the long-term body weight set point in humans.


Severity of hypoglycaemia and health-related quality of life, work productivity and healthcare costs in patients with type 2 diabetes in Europe.
Hypoglycaemia in patients with type 2 diabetes mellitus (T2DM) is associated with poor health outcomes, such as reduced health-related quality of life (HRQoL). This study aimed to assess the impact of hypoglycaemic events by severity on HRQoL, work productivity and healthcare costs in patients with T2DM. European patients with T2DM selected from the National Health and Wellness Survey who were currently receiving pharmacologic therapy were stratified into 3 groups based on the reported history and severity of hypoglycaemic events (no event, nonsevere, severe) experienced in the previous 3 months. Patients' work productivity, HRQoL, healthcare resource use (HCRU) and associated costs were assessed as self-reported outcomes. Of 1269 patients included in the study, 652 (51.4%) patients had not experienced an event, while 533 (42.0%) and 84 (6.6%) patients had experienced nonsevere and severe hypoglycaemic events, respectively, in the previous 3 months. An increase in hypoglycaemia severity was associated with a decrease in HRQoL, and an increase in HCRU and healthcare costs. The impact of hypoglycaemia varies by severity and has a negative impact on HRQoL and overall HCRU and costs.


Serum high density lipoprotein and its relationship to cardiovascular disease risk factor variables in children--the Bogalusa heart study.
Serum high density lipoprotein is increasingly recognized as a negative risk for cardiovascular disease. The distribution and interrelationship of serum lipids, lipoproteins, anthropometric measurements and blood pressures were determined in some 5,000 children. Children had mean +/- S.D. alpha-lipoprotein cholesterol levels (mg/100 ml) of 36 +/- 15 at birth, 51 +/- 22 at 6 mo, 53 +/- 18 at 1 yr, 60 +/- 19 at preschool age (2 1/2-5 1/2 yr) and 68 +/- 22 at school age (5-14 yr), reflecting a sharp increase in alpha-lipoprotein between birth and school-age years, when these levels remained relatively stable through age 14. Although white children tended to have higher levels of total cholesterol and alpha-lipoprotein at birth than black children, during childhood this trend was reversed and the differences were pronounced in school-age children (p less than 0.0001). Unlike in adulthood, boys had slightly higher levels of alpha-lipoprotein than girls. The alpha-lipoprotein was negatively correlated with pre-beta-lipoprotein and to a lesser extent with beta-lipoprotein classes. There was an inverse relationship between alpha-lipoprotein and obesity with a consistently significant relationship (p less than 0.01) in older children (10-14 yr). Children with higher levels of alpha-lipoprotein have lower levels of blood pressure, beta-lipoprotein and a lower obesity index.


Lymphoid crisis with T-cell phenotypes in a patient with Philadelphia chromosome negative chronic myeloid leukaemia.
A case of Philadelphia (Ph1) chromosome negative chronic myeloid leukaemia (CML) developed lymphoid crisis. Immunological marker studies disclosed that the lymphoid cells were sheep erythrocyte-rosetting-, Leu-1+, Leu-5+, OKT-4+, OKT-8+, common ALL antigen-, HLA-DR-, cytoplasmic and surface immunoglobulin-, MAS 036C(antithymocyte)+ (after in vitro culture) and terminal deoxynucleotidyl transferase-, indicating T-cell phenotypes, probably of common thymocytes. Cytochemical staining also demonstrated immature T-cell characters: dot-positivity for acid phosphatase and beta-glucuronidase, and negative for acid alpha-naphthyl acetate esterase. All bone marrow metaphases exhibited normal karyotypes. Our observation suggests that the neoplastic features of a common stem cell for myeloid and lymphoid cell lines are very similar in Ph1 positive and negative CMLs, and that the stem cell can differentiate towards T-lineage.


Hindlimb unweighting induces changes in the RhoA-Rho-kinase pathway of the rat abdominal aorta.
Hindlimb unweighting (HLU) in rats mimics the fluid shift experienced by astronauts and may serve as a model for ground-based orthostatic hypotension. It has been shown that the abdominal aorta of HLU rats exhibits a deficit in contractile response to adrenergic agonists. The hypothesis of the present study was that decreased activity in the RhoA/Rho-kinase pathway could contribute to that deficit. Wistar rats were subjected to 20 days of HLU treatment. Abdominal aorta rings from HLU and control rats were suspended in baths for measurement of contraction. Concentration response curves were obtained to the alpha adrenergic agonist, phenylephrine and the thromboxane-mimetic, U46619. HLU treatment caused decreased contraction in response to both. The Rho-kinase inhibitor, Y27632, caused a reduction in the phenylephrine-induced contraction in control, but not HLU aorta. Other rings were frozen after stimulation 1 microM U46619 or phenylephrine. Western analysis revealed a decreased expression of RhoA, but increased expression of both Rho kinase and MYPT1, the regulatory subunit of myosin light chain (MLC) phosphatase. MYPT1 and MLC phosphorylation was decreased by HLU in phenylephrine stimulated aorta. Decreased activity in the RhoA/Rho-kinase pathway may be involved in the decreased contraction seen in the HLU abdominal aorta.


A pharmacokinetic model for predicting pesticide residues in fish.
By using the compartmental analysis approach of pharmacokinetics, found useful in describing the kinetics of a foreign compound in a biological system, a rather simple mathematical model was developed to predict, by computer simulation, steady-state pesticide residues in fish from limited pre-steady-state experimental data. Such steady-state data are needed in order to calculate the tendency of the residue to concentrate in fish--the so-called bioconcentration factor. The model was applied to four different carbon-14 labeled biocides at seven concentrations. Considering the simplicity of the model and the fact that residues were measured by the non-specific technique of radioassay, model predicted values were in good agreement with experimental values.


A population-based audit for diagnosing colorectal cancer.
Knowledge of the diagnostic work-up of colorectal cancer is a prerequisite to improve its quality. Family history is one of few known risk factors of the disease and it is therefore important to investigate to what extent this factor is used in routine management. Copies of records from all health-care suppliers visited during diagnostic work-up were requested for 227/235 (97%) patients with recently diagnosed colorectal cancer in the county of Västmanland during 1998-99. A first consultation was identified and records and all diagnostic measures related to the initial consultation were scrutinized. A family history of colorectal cancer was known for 179 patients. Most of the patients, 107 (66%) colon and 57 (86%) rectal cancer patients, had consulted with a general practitioner. The median diagnostic work-up time was 42 days (IQ 12-110) for colon and 23 days (IQ 0-49) for rectal cancer. A double-contrast barium enema was the most commonly used diagnostic method for colon cancer. Family history was documented at the first consultation in 2/179 (1%) cases. In patients with right-sided cancer, median diagnostic work-up time was 53 days in patients with a positive result of faecal occult blood test (FOBT) as compared with 448 in patients with a negative result (P < 0.01). Primary care is the key actor in diagnosing rectal cancer. The restricted capacity for X-ray is one of the main obstacles in detection of colon cancer. Family history is rarely documented during diagnostic work-up of colorectal cancer. The benefit of using FOBT in symptomatic patients is questioned.


Cotransport of water by Na⁺-K⁺-2Cl⁻ cotransporters expressed in Xenopus oocytes: NKCC1 versus NKCC2.
The NKCC1 and NKCC2 isoforms of the mammalian Na⁺–K⁺–2Cl⁻ cotransporter were expressed in Xenopus oocytes and the relation between external ion concentration and water fluxes determined.Water fluxes were determined from changes in the oocytes volume and ion fluxes from 86Rb+ uptake. Isotonic increases in external K⁺ concentration elicited abrupt inward water fluxes in NKCC1; the K⁺ dependence obeyed one-site kinetics with a K₀.₅ of 7.5 mM. The water fluxes were blocked by bumetanide, had steep temperature dependence and could proceed uphill against an osmotic gradient of 20 mosmol l⁻¹. A comparison between ion and water fluxes indicates that 460 water molecules are cotransported for each turnover of the protein. In contrast, NKCC2 did not support water fluxes.Water transport in NKCC1 induced by increases in the external osmolarity had high activation energy and was blocked by bumetanide. The osmotic effects of NaCl were smaller than those of urea and mannitol. This supports the notion of interaction between ions and water in NKCC1 and allows for an estimate of around 600 water molecules transported per turnover of the protein. Osmotic gradients did not induce water transport in NKCC2. We conclude that NKCC1 plays a direct role for water balance in most cell types, while NKCC2 fulfils its role in the kidney of transporting ions but not water. The different behaviour of NKCC1 and NKCC2 is discussed on the basis of recent molecular models based on studies of structural and molecular dynamics.


Persons with multiple disabilities use forehead and smile responses to access or choose among technology-aided stimulation events.
A variety of technology-aided programs have been developed to help persons with congenital or acquired multiple disabilities access preferred stimuli or choose among stimulus options. The application of those programs may pose problems when the participants have very limited behavior repertoires and are unable to use conventional responses and microswitches. The present two studies assessed non-conventional response-microswitch solutions for three of those participants. Study I included two participants who were exposed to a program in which forehead skin movement was the response required to access preferred stimulation. The microswitch was an optic sensor combined with a small black sticker on the forehead. Study II included one participant who was exposed to a program in which a smile response was required to choose among stimuli. The microswitch for monitoring the smile was a new camera-based technology. The results of the two studies showed that the response-microswitch solutions were suitable for the participants and enabled them to perform successfully. Implications of the studies for people with limited motor behavior and issues for future research were discussed.


Evaluation of the operative parameters for a human prolactin radioimmunoassay in serum.
A sensitive homologous radioimmunoassay (RIA) for human prolactin (hPRL) was evaluated. The experimental parameters of this double antibody solid phase (DASP)-hPRL-RIA are discribed. The RIA measures concentrations of hPRL as low as 1 ng/ml of human serum and allows repetitive measurements and accurate delineation of serum hPRL. The assay is specific for hRPL; only HGH shows a weak cross-reaction. The antigen recovery tests are quantitative. The serum concentrations of hPRL in a group of randomly selected male and female individuals averaged 6.7 +/- 3.3 ng/ml, respectively. In a chronobiological study performed on 21 subjects (5 male and 16 female), a statistically significant rhythm with a 24-h period was detected.


Precystectomy nomogram for prediction of advanced bladder cancer stage.
To evaluate precystectomy prediction of pT and pN stages at cystectomy. Multivariate logistic regression analyses modelled variables of 726 evaluable patients treated with radical cystectomy and bilateral pelvic lymphadenectomy. The first set of models predicted pT(3-4) stage at cystectomy, and the second set predicted pN(1-3) stages at cystectomy. Transurethral resection (TUR) predictors consisted of 2002 T stage, 1973 WHO tumour grade, presence of carcinoma in situ, age, gender, and delivery of neo-adjuvant chemotherapy. The area under the ROC curve quantified nomogram accuracy. Two hundred bootstrap resamples were used to reduce overfit bias. At TUR, 11% of patients were staged as pT(3-4) versus 42% at cystectomy. Lymph node metastases were found in 24% of patients at cystectomy (pN(1-3)). The multivariate pT(3-4) nomogram was 75.7% accurate versus 71.4% for TUR T stage. The multivariate pN(1-3) nomogram was 63.1% accurate versus 61.0% for TUR T stage. Multivariate nomograms are not perfect, but they do predict more accurately than TUR T stage alone.


Phenotypic and genotypic characteristics of a carbapenem-resistant Serratia marcescens cohort and outbreak: describing an opportunistic pathogen.
Serratia marcescens is an emerging opportunistic pathogen with high genetic diversity. This article describes the microbiological characteristics of isolates and the risk factors for infections caused by carbapenem-resistant S. marcescens. A retrospective study of patients colonized (n=43) and infected (n=20) with carbapenem-resistant S. marcescens over a 3-year period was conducted. Polymerase chain reaction for carbapenemase genes and molecular typing of all available strains was performed. Forty-two isolates were analysed, including three environmental samples identified during an outbreak. Thirty-five carbapenem-resistant S. marcescens carried blaKPC-2, one isolate was blaNDM-positive and four isolates carried blaOXA-101. The genomes were grouped into three clusters with 100% bootstrap; three patterns of mutations on ompC and ompF were found. The strains carried virulence genes related to invasion and haemolysis, and the environmental strains presented fewer mutations on the virulence genes than the clinical strains. Multi-variate analysis showed that previous use of polymyxin (P=0.008) was an independent risk factor for carbapenem-resistant S. marcescens infection. This study highlighted that blaKPC-2 in association with ompC or ompF mutation was the most common mechanism of resistance in the study hospital, and that previous use of polymyxin was an independent risk factor for carbapenem-resistant S. marcescens. There was a predominant clone, including the environmental isolates, suggesting that cross-transmission was involved in the dissemination of this pathogen.


The role of α9β1 integrin and its ligands in the development of autoimmune diseases.
Adhesion of cells to extracellular matrix proteins through integrins expressed on the cell surface is important for cell adhesion/motility, survival, and differentiation. Recently, α9β1 integrin was reported to be important for the development of autoimmune diseases including rheumatoid arthritis, multiple sclerosis, and their murine models. In addition, ligands for α9β1 integrin, such as osteopontin and tenascin-C, are well established as key regulators of autoimmune diseases. Therefore, this review focused on the role of interactions between α9β1 integrin and its ligands in the development of autoimmune diseases.


The role of maternal age, growth, and environment in shaping offspring performance in an aerial conifer seed bank.
Maternal effects have been demonstrated to affect offspring performance in many organisms, and in plants, seeds are important mediators of these effects. Some woody plant species maintain long-lasting canopy seed banks as an adaptation to wildfires. Importantly, these seeds stored in serotinous cones are produced by the mother plant under varying ontogenetic and physiological conditions. We sampled the canopy seed bank of a highly serotinous population of Pinus pinaster to test whether maternal age and growth and the environmental conditions during each crop year affected seed mass and ultimately germination and early survival. After determining retrospectively the year of each seed cohort, we followed germination and early survival in a semi-natural common garden. Seed mass was related to maternal age and growth at the time of seed production; i.e., slow-growing, older mothers had smaller seeds, and fast-growing, young mothers had larger seeds, which could be interpreted either as a proxy of senescence or as a maternal strategy. Seed mass had a positive effect on germination success, but aside from differences in seed mass, maternal age had a negative effect and diameter had a positive effect on germination timing and subsequent survival. The results highlight the importance of maternal conditions combined with seed mass in shaping seedling establishment. Our findings open new insights in the offspring performance deriving from long-term canopy seed banks, which may have high relevance for plant adaptation.


Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava.
Plant virus vectors designed for virus-mediated protein overexpression (VOX), virus-induced gene silencing (VIGS), and genome editing (VIGE) provide rapid and cost-effective tools for functional genomics studies, biotechnology applications and genome modification in plants. We previously reported that a cassava common mosaic virus (CsCMV, genus Potexvirus)-based VIGS vector was used for rapid gene function analysis in cassava. However, there are no VOX and VIGE vectors available in cassava. In this study, we developed an efficient VOX vector (CsCMV2-NC) for cassava by modifying the CsCMV-based VIGS vector. Specifically, the length of the duplicated putative subgenomic promoter (SGP1) of the CsCMV CP gene was increased to improve heterologous protein expression in cassava plants. The modified CsCMV2-NC-based VOX vector was engineered to express genes encoding green fluorescent protein (GFP), bacterial phytoene synthase (crtB), and Xanthomonas axonopodis pv. manihotis (Xam) type III effector XopAO1 for viral infection tracking, carotenoid biofortification and Xam virulence effector identification in cassava. In addition, we used CsCMV2-NC to deliver single guide RNAs (gMePDS1/2) targeting two loci of the cassava phytoene desaturase gene (MePDS) in Cas9-overexpressing transgenic cassava lines. The CsCMV-gMePDS1/2 efficiently induced deletion mutations of the targeted MePDS with the albino phenotypes in systemically infected cassava leaves. Our results provide a useful tool for rapid and efficient heterologous protein expression and guide RNA delivery in cassava. This expands the potential applications of CsCMV-based vector in gene function studies, biotechnology research, and precision breeding for cassava.


Synthesis, Characterization, and Biological Activities of Pendant Arm-Pyridyltetrazole Copper(II) Complexes: DNA Binding/Cleavage Activity and Cytotoxic Studies.
2-(1H-Tetrazol-5-yl)pyridine (L) has been reacted separately with Me2NCH2CH2Cl⋅HCl and ClCH2CH2OH to yield two regioisomers in each case, N,N-dimethyl-2-[5-(pyridin-2-yl)-1H-tetrazol-1-yl]ethanamine (L1)/N,N-dimethyl-2-[5-(pyridin-2-yl)-2H-tetrazol-2-yl]ethanamine (L2) and 2-[5-(pyridin-2-yl)-1H-tetrazol-1-yl]ethanol (L3)/2-[5-(pyridin-2-yl)-2H-tetrazol-2-yl]ethanol (L4), respectively. These ligands, L1-L4, have been coordinated with CuCl2 ⋅H2O in 1 : 1 composition to furnish the corresponding complexes 1-4. EPR Spectra of Cu complexes 1 and 3 were characteristic of square planar geometry, with nuclear hyperfine spin 3/2. Single X-ray crystallographic studies of 3 revealed that the Cu center has a square planar structure. DNA binding studies were carried out by UV/VIS absorption; viscosity and thermal denaturation studies revealed that each of these complexes are avid binders of calf thymus DNA. Investigation of nucleolytic cleavage activities of the complexes was carried out on double-stranded pBR322 circular plasmid DNA by using a gel electrophoresis experiment under various conditions, where cleavage of DNA takes place by oxidative free-radical mechanism (OH(⋅)). In vitro anticancer activities of the complexes against MCF-7 (human breast adenocarcinoma) cells revealed that the complexes inhibit the growth of cancer cells. The IC50 values of the complexes showed that Cu complexes exhibit comparable cytotoxic activities compared to the standard drug cisplatin.


[Approach and complications associated with suburethral synthetic slings in women: Systematic review and meta-analysis].
We aim to assess the complications associated with different approaches used in female suburethral sling surgery. We performed a research on Medline using the following keywords: "suburethral slings", "complications", "safety" and "randomized". Only randomized clinical trials including women and reporting intra- and postoperative complications associated with the retropubic (RP) approach; TOT and/or TVT-O were included. The meta-analysis was conducted using the Review Manager (RevMan 5.3) software delivered by the "Cochrane Library". Out of 176 articles, 23 were included in synthesis. Risks of bladder perforation during surgery (60/1482 vs 5/1479; OR=6.44; 95% CI [3.32-12.50]) and postoperative urinary retention (48/1160 vs 24/1159; OR=1.93; 95% CI [1.26-3.12]) were significantly higher with the RP approach, when compared with the transobturator (TO) approach (TOT or TVT-O). Conversely, the risk of prolonged postoperative pain was significantly lower after RP approach, when compared with TO approach (24/1156 vs 69/1149; OR=0.36; 95% CI [0.23-0.56]). Risks of intraoperative urethral injury, postoperative erosion and de novo overactive bladder were comparable between the two approaches. Data regarding the comparison between TOT and TVT-O were scarce and did not allow us to conclude about complications associated with. The RP approach was associated with a significant risk of bladder perforation and postoperative urinary retention. The TO approach was associated with a higher risk of prolonged postoperative pain.


Multiple electrocardiographic anomalies during anaesthesia in an athlete.
A 22-year-old athlete was scheduled for a minor surgical procedure under general anaesthesia. During anaesthesia, his electrocardiogram demonstrated multiple episodes of dysrhythmias including complete bundle branch block, atrioventricular (AV) block, isorhythmic atrioventricular dissociation with junctional rhythm. Administration of atropine 1.0 mg IV terminated the last episode of dysrhythmias. Postoperatively, a resting 12-lead electrocardiogram showed first degree AV block, ST-segment elevation and prominent U waves. A 24 hour Holter recording demonstrated first degree atrioventricular block, episodes of marked sinus arrhythmias and one episode of sinus tachycardia at a rate of 152 beats X min-1. Treadmill stress testing revealed peak achieved heart rate of 200 beats X min-1 without ischaemia. These findings collectively indicated athletic heart syndrome. Implications of athletic heart syndrome for the anaesthetist are reviewed and discussed.


Fertilization rates using intracytoplasmic sperm injection are greater than subzonal insemination but are dependent on prior treatment of sperm.
To determine a suitable method of sperm preparation for use in micromanipulation. To compare the fertilization rates of sibling oocytes inseminated by intracytoplasmic sperm injection (ICSI) and subzonal insemination (SUZI). Two methods of gamete micromanipulation to overcome male factor infertility were compared. Within this study, three trials were conducted to determine the most suitable method of sperm preparation. This method then was used to complete the study. Procedures were performed in a teaching hospital research environment. Ninety-six patients were recruited for this study, undergoing 99 stimulation cycles. Sibling oocytes were inseminated by subzonal sperm or intracytoplasmic injection. Fertilization, zygote development, and pregnancy rates. In the absence of manipulative pretreatment of sperm (trial 1), there was no difference in normal fertilization rates between ICSI and SUZI (19% and 25%, respectively). In the second trial there again was no sperm pretreatment for SUZI but, for ICSI, a polyvinylpyrrolidone (PVP) solution was used to reduce sperm velocity and the sperm tails were incised before injection. The fertilization rates were significantly different between ICSI (44%) and SUZI (17%). In the third trial, PVP was added to the sperm used for both types of insemination and the sperm tails also were incised for the ICSI insemination. Fertilization was again significantly different (16% for SUZI and 44% for ICSI). The trial 2 method of sperm preparation then was used to complete the study (trial 4) and confirmed the results of trial 2 (21% for SUZI and 42% for ICSI). The results suggest that ICSI can give improved fertilization compared with SUZI provided the sperm are treated before injection. No significant differences were found in the development rates of zygotes suitable for transfer or cryopreservation between the two micromanipulation methods.


A prospective, randomized study comparing highly selective vagotomy and extended highly selective vagotomy in patients with duodenal ulcer.
The recurrence rate of duodenal ulcer after highly selective vagotomy is nearly 10 percent. To diminish this percentage, extended highly selective vagotomy with sectioning the gastroepiploic nerves has been proposed in order to reduce postoperative gastric acid secretion. We have prospectively compared the decrease in gastric acid secretion through measurement of basal acid output, maximal acid output, and peak acid output in patients who underwent highly selective vagotomy or extended highly selective vagotomy. No significant differences in postoperative gastric acid secretion were found and, therefore, no changes in the probability of postoperative recurrence of duodenal ulcer were seen.


Molecular diagnosis of genodermatoses.
The progress of molecular genetics helps clinicians to prove or exclude a suspected diagnosis for a vast and yet increasing number of genodermatoses. This leads to precise genetic counselling, prenatal diagnosis and preimplantation genetic haplotyping for many inherited skin conditions. It is also helpful in such occasions as phenocopy, late onset and incomplete penetrance, uniparental disomy, mitochondrial inheritance and pigmentary mosaicism. Molecular methods of two genodermatoses are explained in detail, i.e. genodermatoses with skin fragility and neurofibromatosis type 1.


Lysine-Specific Demethylase 1 in Energy Metabolism: A Novel Target for Obesity.
Obesity develops from an imbalance of energy homeostasis and is associated with the development of metabolic disorders, including insulin resistance and type 2 diabetes. Identification of the underlying molecular mechanisms and effective therapeutic approaches is highly needed. Lysine-specific demethylase 1 (LSD1), an flavin adenine dinucletide-dependent amine oxidase, is implicated in a wide variety of biological processes, including tumorigenesis, stem cell fate decisions, and embryonic development. Recent studies have suggested a vital role of LSD1 in regulating adaptive thermogenesis, mitochondrial biogenesis, glucose, and lipid metabolism. More recently, LSD1 activity was found to be regulated by nutrients, energy status, and hormonal signals, suggesting that it may act as a novel sensor for nutritional regulation of metabolic health. Here, we first discuss the effects of LSD1 on physiological phenotypes, including body weight, fat mass, body temperature, and glucose homeostasis. We also summarize recent understanding of the physiological roles and underlying mechanisms of LSD1 in controlling metabolic functions of adipose and other tissues. Hopefully, a better understanding of the roles of LSD1 in metabolic regulation may provide new perspectives for the nutritional prevention and treatment of obesity.


Challenges faced with opioid prescriptions in the community setting - Australian pharmacists' perspectives.
Prescription opioids (POs) are well recognised for their role in pain management. However over recent years, use of POs has become an increasingly complex public health issue, with the emergence of increasing quantities of POs being misused and abused. Pharmacists represent the 'gatekeepers' of medicines, which can be a challenging role, relating in particular to POs. This study aimed to gain insight into the challenges Australian community pharmacists experience in the dispensation of POs. Sydney Australia. Twenty-five pharmacists were recruited from suburbs in and around Sydney, Australia. Pharmacists were interviewed using an in-depth, semi-structured protocol. All interviews were audio-recorded, transcribed and thematically analysed. Identification of issues/factors that may influence the dispensing of POs. Pharmacists were reportedly confronted with several issues in the dispensing of POs, particularly in relation to the patient and prescriber. Pharmacists reported some individuals becoming increasingly more "creative" in the methods used to obtain POs, rendering detection of potential abuse/misuse increasingly difficult. Poor professional relationships with prescribers-an apparent power dynamic between the two professions, and limited engagement with patients were issues also identified. The majority of participants suggested that an electronic monitoring database would help in the identification of PO abuse/misuse. Education is required to emphasise the importance of inter-professional collaboration between pharmacists and prescribers, as is empathy in the pharmacist-patient relationship. Prescription drug monitoring programs and prescribers sending electronic prescriptions directly to the pharmacist were perceived as helpful initiatives to undertake.