Synthesis and evaluation of a new vesamicol analog o-[(11)C]methyl-trans-decalinvesamicol as a PET ligand for the vesicular acetylcholine transporter.
We focused on the vesicle acetyl choline transporter (VAChT) as target for early diagnosis of Alzheimer's diseases because the dysfunction of the cholinergic nervous system is closely associated with the symptoms of AD, such as problem in recognition, memory, and learning. Due to its low binding affinity for the sigma receptors (σ-1 and σ-2), o-methyl-trans-decalinvesamicol (OMDV) demonstrated a high binding affinity and selectivity for vesicular acetyl choline transporter (VAChT). [(11)C]OMDV was prepared and investigated the potential as a new PET ligand for VAChT imaging through in vivo evaluation. [(11)C]OMDV was prepared by a palladium-promoted cross-coupling reaction using [(11)C]methyl iodide, with a radiochemical yield of 60-75%, a radiochemical purity of greater than 98%, and a specific activity of 5-10 TBq/mmol 30 min after EOB. In vivo biodistribution study of [(11)C]OMDV in blood, brain regions and major organs of rats was performed at 2, 10, 30 and 60 min post-injection. In vivo blocking study and PET-CT imaging study were performed to check the binding selectivity of [(11)C]OMDV for VAChT. In vivo studies demonstrated [(11)C]OMDV passage through the blood-brain barrier (BBB) and accumulation in the rat brain. The regional brain accumulation of [(11)C]OMDV was significantly inhibited by co-administration of vesamicol. In contrast, brain accumulation of [(11)C]OMDV was not significantly altered by co-administration of (+)-pentazocine, a selective σ-1 receptor ligand, or (+)-3-(3-hydroxyphenyl)-N-propylpiperidine [(+)-3-PPP], a σ-1 and σ-2 receptor ligand. PET-CT imaging revealed inhibition of [(11)C]OMDV accumulation in the brain by co-administration of vesamicol. [(11)C]OMDV selectively binds to VAChT with high affinity in the rat brain in vivo, and that [(11)C]OMDV may be utilized in the future as a specific VAChT ligand for PET imaging.


Quinic acid esters from Erycibe obtusifolia with antioxidant and tyrosinase inhibitory activities.
AbstractA new quinic acid derivative, 3-O-syringoylquinic acid methyl ester (1), along with eight known quinic acid derivatives (2-9), three coumarins (10-12), one phenylpropanoid (13), three feruloyltyramine derivatives (14-16), one lignan (17) and two isoflavones (18-19) were isolated from an ethyl acetate-soluble fraction of the roots and stems of Erycibe obtusifolia. The structure was elucidated on the basis of spectroscopic methods such as 1D and 2D-NMR, including HR-ESI-MS spectrometry. All of these compounds were investigated for their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and inhibitory effects on mushroom tyrosinase. Compounds 2-9, quinic acid derivatives with caffeoyl moiety, showed significant DPPH radical scavenging activity. Moreover, compounds 2 and 5-10 showed weak mushroom tyrosinase inhibitory effects.


Familial DHCR7 genotype presenting as a very mild form of Smith-Lemli-Opitz syndrome and lethal holoprosencephaly.
Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive metabolic disorder caused by variants in the DHCR7 gene. In cholesterol biosynthesis, 7-dehydrocholesterol (7-DHC) is converted to cholesterol by the enzyme 7-DHC reductase, which is encoded by the gene DHCR7. Thus, an elevated 7-DHC is indicative of SLOS. Characteristically SLOS is usually associated with congenital anomalies, dysmorphisms, and moderate to severe neurodevelopmental delay. However, there are rare descriptions of individuals with milder phenotypes. We report a mild case of SLOS presenting with short stature, cleft palate, imperforate anus, and mild language delay with subtle dysmorphic features. 7-DHC was not elevated at 1 year of age and SLOS considered excluded at this time. The parents had two pregnancies with holoprosencephaly. Whole exome sequencing of one of the fetuses identified compound heterozygous pathogenic variants in the DHCR7 gene (c.964-1G>C (p.?) and c.1039G>A (p.Gly347Ser) causative of SLOS. The proband with a mild form of SLOS was also found to have the same DHCR7 variants as the fetus and repeat testing of 7-DHC at 4 years of age was elevated, in keeping with SLOS. This case is the first to describe a wide intrafamilial phenotypic spectrum of SLOS as a result of the same DHCR7 genotype. This case also supports the findings of others that a normal or near normal development should not exclude SLOS. As demonstrated in this case exclusion of a metabolic diagnosis because of a negative biochemical marker such as 7-DHC is not absolute and if clinical suspicion remains genomic sequencing is warranted.


Stereotactic body radiation therapy in the treatment of multiple primary lung cancers.
Patients with multiple primary lung cancers (MPLC) present a therapeutic dilemma, particularly when they are at high risk for surgical resection. We evaluated the role of stereotactic body radiation therapy (SBRT) in the treatment of MPLC. A prospective thoracic SBRT registry was explored for patients with either synchronous or metachronous MPLC treated with SBRT for one or both of their tumors. Sixty-three patients were identified and clinical data were analyzed. Fifteen patients had synchronous lesions and 48 patients had metachronous lesions. Seventy-six lesions were treated with SBRT. Median follow-up was 24 months for living patients. Median progression-free survival (PFS) and overall survival (OS) for the entire cohort was 15.5 and 20 months, respectively. Patients with metachronous MPLC had a significantly higher 2 year PFS (53.3% vs. 0%, p=0.0466) compared to patients with synchronous MPLC. Likewise, 2 year OS was also superior for patients with metachronous versus synchronous MPLC (68.1% vs. 27.5%, p=0.0014). Six tumors (7.9%) recurred within the radiation field. There were no grade ≥3 toxicities. SBRT for patients with MPLC appears to be a safe and effective local treatment alternative to surgery, particularly for medically inoperable patients. Patients with metachronous MPLC have encouraging survival rates, and thus local therapy appears justified. However, patients with synchronous MPLC have poor OS and PFS despite having excellent local control, and thus the utility of local therapy in this population requires further study.


Microglial dysfunction connects depression and Alzheimer's disease.
Alzheimer's disease (AD) and major depressive disorder (MDD) are highly prevalent neuropsychiatric conditions with intriguing epidemiological overlaps. Depressed patients are at increased risk of developing late-onset AD, and around one in four AD patients are co-diagnosed with MDD. Microglia are the main cellular effectors of innate immunity in the brain, and their activation is central to neuroinflammation - a ubiquitous process in brain pathology, thought to be a causal factor of both AD and MDD. Microglia serve several physiological functions, including roles in synaptic plasticity and neurogenesis, which may be disrupted in neuroinflammation. Following early work on the 'sickness behavior' of humans and other animals, microglia-derived inflammatory cytokines have been shown to produce depressive-like symptoms when administered exogenously or released in response to infection. MDD patients consistently show increased circulating levels of pro-inflammatory cytokines, and anti-inflammatory drugs show promise for treating depression. Activated microglia are abundant in the AD brain, and concentrate around senile plaques, hallmark lesions composed of aggregated amyloid-β peptide (Aβ). The Aβ burden in affected brains is regulated largely by microglial clearance, and the complex activation state of microglia may be crucial for AD progression. Intriguingly, recent reports have linked soluble Aβ oligomers, toxins that accumulate in AD brains and are thought to cause memory impairment, to increased brain cytokine production and depressive-like behavior in mice. Here, we review recent findings supporting the inflammatory hypotheses of AD and MDD, focusing on microglia as a common player and therapeutic target linking these devastating disorders.


Changes in some pregnancy biomarkers of Yankasa ewes experimentally infected with Trypanosoma evansi.
The study was designed to determine the effect of Trypanosoma evansi infection on some pregnancy biomarkers of Yankasa ewes (YE). Twenty pregnant YE were assigned into 3 groups (A, B and C) comprising 7 ewes each in groups A and B, while group C comprise 6 YE. Groups A and B were each inoculated with blood containing approximately 1.0 × 10(6) of T. evansi through the jugular vein on days 59 and 110 of pregnancy, representing second and third trimesters, respectively, while group C served as the uninfected control. Progesterone (P4) and pregnancy specific protein-B (PSPB) of YE in group A were significantly (p < 0.05) high at weeks 4 and 12 post infection (pi) respectively, while there was no significant (p > 0.05) difference in P4 and PSPB of YE in groups B. Estrone sulfate (E1S) significantly (p < 0.05) decrease for YE in group A at weeks 2 and 11 pi. However, it was not significantly (p > 0.05) different in group B. Cortisol concentration of YE in group A was significantly (p < 0.05) decreased at week 12 pi. Conversely, the cortisol concentration of YE in group B significantly (p < 0.05) increased at week 3 pi. There was no significant (p > 0.05) association among the pregnancy biomarkers of YE in groups A and B throughout the study, except between progesterone and cortisol in group B, which were significantly associated (r = 0.77, p < 0.05). It was therefore concluded that T. evansi infection affects pregnancy biomarkers more at mid pregnancy than at late pregnancy.


Leishmanicidal effect of curcumin in vitro.
From a study to find anti-parasitic agents from natural resources, we found that curcumin showed the cytotoxicity against leishmania in vitro. The LD50 value of this activity was 37.6+/-3.5 microM.


Pressure distension stimulates the expression of endothelial adhesion molecules in the human saphenous vein graft.
Mechanical trauma occurring during saphenous vein graft harvesting plays a major role in graft failure after coronary bypass surgery. There is increasing evidence that neutrophil-endothelial interaction is involved in the pathogenesis of early graft occlusion. This study evaluates the effect of pressure distension on the expression of endothelial adhesion molecules in human saphenous vein. Segments of saphenous vein graft (SVG) were collected from 20 patients undergoing coronary bypass surgery. We evaluated the expression of intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), and P-selectin on SVG endothelium under basal conditions and after pressure distension at 300 mm Hg. In the same experimental setting we also evaluated adhesion of both unstimulated and activated neutrophils to the endothelium of SVG. Control endothelial cells exhibited only a weak staining for intercellular adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), and P-selectin, whereas the levels of adhesion molecules increased significantly in the distended veins. Similarly, significantly greater adhesion of both unstimulated and activated neutrophils was observed in distended veins compared with control veins. Pressure distension of SVG before coronary bypass surgery induces upregulation of endothelial adhesion molecules, with subsequent increase in neutrophil adhesion to the endothelium. Neutrophil adhesion to endothelial cells may contribute to early failure of SVG.


Gastric acid secretion in the lizard. Ionic requirements and effects of inhibitors.
1. The effects of ion substitution and various inhibitors on the transmucosal potential, short circuit current, mucosal resistance and acid secretion of the lizard gastric mucosa, incubated in an Ussing chamber, have been determined. 2. Ion substitution experiments indicate that the serosal potential step consists of a combined C1- and K+ diffusion potential, and that the mucosal potential step is Na+ dependent and behaves primarily as a Na+ diffusion potential. 3. Experiments with ouabain indicate that the major (Na+, K+)-ATPase activity responsible for maintenance of cation gradients is located on the serosal side of the mucosal cells, and that this pump activity is non-electrogenic. 4. Experiments with amiloride indicate that a passive sodium influx on the mucosal side is essential for the maintenance of the transmucosal potential and short circuit current. 5. Acid secretion requires the presence of sodium and chloride on the serosal side and the maintenance of a high intracellular potassium level through the (Na+, K+)-ATPase system. 6. The effects of acetazolamide and thiocyanate are compatible with an involvement of carbonic anhydrase and anion-dependent ATPase in acid secretion. 7. Upon initiation of acid secretion the serosal membrane permeability for chloride increases and that for potassium decreases.


Relation of spontaneous passage of ureteral calculi to size.
A statistical analysis of 520 ureteral calculi was undertaken to elucidate the correlation between sizes of calculi and the probability of spontaneous passage. The mean length and width of 286 calculi passed spontaneously were 6.3 +/- 2.5 (S.D.) and 4.0 +/- 1.5 mm., respectively; those of 219 calculi removed surgically were 11.7 +/- 5.0 and 7.1 +/- 2.8 mm., respectively, the difference between the two groups being statistically significant. The rate of spontaneous passage of stones within one year after diagnosis was analyzed in relation to sizes of stones which were grouped at successive 1-mm. intervals both in length and width, the over-all rate of spontaneous passage being 53 per cent. It was concluded that stones larger than 8 mm. in width on x-ray films should be removed surgically, while in those smaller than 8 mm. in width higher chances of spontaneous passage by expectant treatment would be anticipated.


Insulin inhibits inducible nitric oxide synthase in skeletal muscle cells.
Recent studies have shown that cytokines and endotoxins impair insulin-stimulated glucose transport by activating the expression of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) production in skeletal muscle cells. In this study, we investigated whether iNOS induction is modulated by insulin in L6 myocytes. Long term exposure of muscle cells to tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) greatly increased iNOS mRNA expression and NO production. Addition of insulin to the cytokine/LPS-treated muscle cells reduced (by approximately 40%) NO production. This inhibition was similar to that observed with the synthetic glucocorticoid dexamethasone, a known inhibitor of iNOS in several cell types. The combination of insulin and dexamethasone was more effective than either agent alone in reducing NO production. Dexamethasone greatly inhibited the effect of cytokines/LPS to induce cellular iNOS mRNA expression. In strong contrast, insulin failed to reduce iNOS mRNA expression under similar conditions. These results show that insulin is a novel inhibitor of iNOS-mediated NO production in skeletal muscle cells. Furthermore, our data indicate that unlike glucocorticoids, insulin does not inhibit NO production by suppression of iNOS gene transcription.


Activities to promote empathy for patients among pharmacy learners: A scoping review.
Provider empathy has been shown to be directly linked to improved patient outcomes. The objective of this scoping review of the literature was to identify and describe learning activities that promote empathy for patients among pharmacy learners. This scoping review was conducted using the following inclusion criteria: publication in English, activities conducted in any academic pharmacy training program (professional degree program, experiential, residency, or fellowship), description of the learning activity(ies) provided, and focus on the experience of empathy/caring/compassion for patients, either human or animal. Articles were excluded if they focused only on skills such as empathic responding or if they did not describe the learning experience. All study designs other than reviews were included. The scoping review revealed 89 full-text articles that met the inclusion criteria. Included studies demonstrated a wide variety of approaches to the design of learning experiences as well as methods of measurement of empathy. Various types of learning modalities have been used to develop empathy in pharmacy learners, with reflection being the most common. A large proportion of studies that assessed empathy development used quasi-experimental or qualitative designs and did not report tests of statistical significance, which would make it difficult to compare the effectiveness of the different learning activities. A variety of approaches have been used among pharmacy learners to develop empathy for patients. Due to the high level of variability in approaches, more rigorous studies are needed to assess the effectiveness of these learning activities.


Screening serum biomarker of knee osteoarthritis using a phage display technique.
To screen serum biomarker of knee osteoarthritis (OA) using a phage random peptide library. A phage random peptide library of random peptide 12-mers was screened with purified immunoglobulin G (IgG) from sera of knee OA patients. Patients with knee rheumatoid arthritis (RA), hip OA, non-erosive hand OA or erosive hand OA, and healthy volunteers were used as controls. A phage clone with inserted peptide TGLESGHGPGDS (named KOA1) showed 90% positive reaction rate with the knee OA patients, significantly higher than that with the knee RA patients (27.8%), the non-erosive hand OA patients (34.3%), the erosive hand OA patients (31.3%) and the healthy controls (12.0%), but not the hip OA patients (82.5%). The novel knee OA mimic peptide KOA1 identified with a random phage display peptide library and sera from knee OA patients could be a potential serum biomarker for knee OA.


[Experimental investigations on the effect of the transducer-specimen distance on the ultrasound attenuation of formalin-fixed testicular tissue].
Additional information on the tissue structure are possible by estimation of sound physical parameters in comparison with normal B-scan ultrasonography, especially the frequency-dependent ultrasound attenuation. The ultrasound attenuation measurements were made on histochemical normal and pathological testicular tissue with changing transducer-specimen distance. No markedly difference in damaging behaviour was found between normal and pathological testicular tissue at different distances.


Improving the efficiency of 4A-zeolite synthesized from kaolin by amine functionalization for CO2 capture.
This study focuses on optimizing the CO2 adsorption capacity of 4A-zeolite synthesized from kaolin by employing structural modifications through impregnation with tetraethylenepentamine (TEPA) and diethanolamine (DEA). Various analytical techniques were utilized to evaluate the effectiveness of these modifications. Design expert software and response surface methodology (RSM) was employed for data analysis and operational variable optimization, leading to improved CO2 adsorption performance of the modified zeolites. The adsorption capacity of the modified zeolites was assessed under different temperatures, pressures, and amine concentrations using a test device. The optimal adsorption capacity of 4A-DEA adsorbent is found to be 579.468 mg/g, with the optimal operational variables including a temperature of 25.270 °C, pressure of 8.870 bar, and amine concentration of 11.112 wt%. The analysis shows that the adsorption process involves both physisorption and chemisorption, and the best kinetic model is the fractional-factor model.


[Hypertension--follow up in a rural practice].
In a rural general practitioner's office, 286 patients with all WHO stages of hypertension, were followed up. It was found that hypertensive drugs are more commonly employed in daily practice as basal treatment among the general population. This treatment complies with the requirements of clinical medicine with respect to frequency, patient age, duration of treatment, effectiveness and drug combinations.


Gastric peroral endoscopic myotomy outcomes after 4 years of follow-up in a large cohort of patients with refractory gastroparesis (with video).
Gastric peroral endoscopic myotomy (G-POEM) is used for refractory gastroparesis (RG) with good early-term but variable mid- and long-term outcomes. Limited data exist about candidates and long-term clinical and predictive factors. Our aim was to evaluate the 4-year follow-up efficacy and predictive factors in patients with RG. Confirmed RG patients were included and evaluated between April 2017 and December 2021. Gastroparesis Cardinal Symptom Index (GCSI) score, retention percentage at 4 hours (RP4H), mean half-emptying time (MHET), and the 36-item short-form survey (SF-36) were performed at 1, 6, 12, 18, 24, 30, 36, 42, and 48 months. After G-POEM, 374 patients with RG were included: 141 patients (37.7%) had diabetic gastroparesis (DG), 115 (30.7%) had idiopathic gastroparesis (IG), 102 (27.3%) had postsurgical gastroparesis (PSG), and 16 (4.3%) had other etiologies. After the 48-month evaluation, 102 patients completed follow-up (DG, 58; IG, 22; PSG, 18; other, 4). Before G-POEM, GCSI score, RP4H, and MHET were 3.84 ± .53, 44% (interquartile range [IQR], 11-68), and 246 minutes (IQR, 150-368), respectively, and after the 48-month evaluation improved to 2.1 ± .70 (P < .001), 15.5% (IQR, 0-36; P = .021), and 135 minutes (IQR, 67-290; P = .045), respectively. At the 48-month evaluation, clinical success was 77.5%. DG showed the best outcomes (DG vs IG vs PSG vs other: 86.5% vs 72.5% [P = .001] vs 72.1% [P = .003] vs 68.8% [P < .001]). Long-term success predictors were DG (odds ratio [OR], 5.113; 95% confidence interval [CI], 1.643-5.981; P = .035), early diagnosis (OR, 2.455; 95% CI, 1.129-3.522; P = .042), nausea/vomiting (OR, 3.541; 95% CI, 1.881-5.511; P = .012), GCSI score at 6 months (1.5-2) (OR, 3.612; 95% CI, 2.122-5.317; P = .022), and RP4H <10% at 6 months (OR, 2.188; 95% CI, 1.435-4.233; P = .039). G-POEM is an effective 4-year treatment in patients with RG, especially in DG, establishing a potential first-line therapy in these patients. However, randomized controlled clinical trials are needed to confirm these results. (Clinical trial registration number: NTC03126513.).


Serum metabolomics of hyperbilirubinemia and hyperuricemia in the Tibetan plateau has unique characteristics.
Few studies have provided data on the metabolomics characteristics of metabolic diseases such as hyperuricemia and hyperbilirubinemia in the Tibetan plateau. In the current study, we sought to investigate the serum metabolomics characteristics of hyperbilirubinemia and hyperuricemia in the Tibetan plateau, with the aim to provide a basis for further research on their pathogenesis, prevention, and treatment. The study participants were born in low-altitude areas below 1000 m and had no prior experience living in a high-altitude area before entering Golmud, Tibet (average elevation: 3000 m) and Yushu, Qinghai (average elevation: 4200 m). Thirty-four participants with hyperbilirubinemia (18 in Golmud and 16 in Yushu), 24 participants with hyperuricemia, and 22 healthy controls were enrolled. The serum samples of subjects were separated and then sent to a local tertiary hospital for biochemical examination. Serum widely targeted technology, based on the ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) platform, was used to detect serum metabolites and differential metabolites. Compared to the healthy controls, hyperbilirubinemia patients from Golmud showed 19 differential metabolites, hyperbilirubinemia patients from Yushu showed 12 differential metabolites, and hyperuricemia patients from Yushu showed 23 differential metabolites. Compared to the hyperbilirubinemia patients from Golmud that is at a low altitude, the Yushu groups had 33 different metabolites. Differential metabolites are primarily classified into amino acids and their derivatives, nucleotides and their derivatives, organic acids and their derivatives, and lipids/fatty acids. These are related to metabolic pathways such as caffeine metabolism, arachidonic acid metabolism, and tyrosine metabolism. Hyperbilirubinemia and hyperuricemia in the Tibetan plateau have unique serum metabolomics characteristics. Glycine derivatives and arachidonic acid and its derivatives were associated with plateau hyperbilirubinemia, and vanillic acid and pentadecafluorooctanoic acid were associated with plateau hyperuricemia.


ABCG2-overexpressing S1-M1-80 cell xenografts in nude mice keep original biochemistry and cell biological properties.
S1-M1-80 cells, derived from human colon carcinoma S1 cells, are mitoxantrone-selected ABCG2-overexpressing cells and are widely used in in vitro studies of multidrug resistance(MDR). In this study, S1-M1-80 cell xenografts were established to investigate whether the MDR phenotype and cell biological properties were maintained in vivo. Our results showed that the proliferation, cell cycle, and ABCG2 expression level in S1-M1-80 cells were similar to those in cells isolated from S1-M1-80 cell xenografts (named xS1-M1-80 cells). Consistently, xS1-M1-80 cells exhibited high levels of resistance to ABCG2 substrates such as mitoxantrone and topotecan, but remained sensitive to the non-ABCG2 substrate cisplatin. Furthermore, the specific ABCG2 inhibitor Ko143 potently sensitized xS1-M1-80 cells to mitoxantrone and topotecan. These results suggest that S1-M1-80 cell xenografts in nude mice retain their original cytological characteristics at 9 weeks. Thus, this model could serve as a good system for further investigation of ABCG2-mediated MDR.


Catalytic-independent inhibition of cIAP1-mediated RIP1 ubiquitination by EGLN3.
EGLN3 belongs to the EGLN family of prolyl hydroxylases that are able to catalyze the hydroxylation of proteins such as the α subunits of hypoxia-inducible factor. We and others have shown that EGLN3 negatively regulates the canonical NFκB pathway. Mechanistically, we demonstrated that EGLN3 inhibits ubiquitination of IKKγ (the regulatory subunit of IκB kinase complex) which is vitally important for NFκB activation. Polyubiquitination of the RIP1 (receptor-interacting protein 1) kinase is important for NFκB activation triggered by tumor necrosis factor α. It remains to be determined whether EGLN3 is able to modulate RIP1 ubiquitination catalyzed by cIAP1 (cellular inhibitor of apoptosis protein 1). This study shows that EGLN3 interacts with cIAP1 and suppresses cIAP1-mediated RIP1 ubiquitination via the C-terminal region. The hydroxylase activity is not required for the ability of EGLN3 to restrain RIP1 ubiquitination. Furthermore, EGLN3 is a novel binding protein of RIP1. The C-terminal region of EGLN3 is responsible for its interaction with RIP1. EGLN3 hydroxylase activity is not essential for the EGLN3-RIP1 interaction. EGLN3 interferes with the association between RIP1 and cIAP1, and attenuates RIP1-induced NFκB activation. This study provides novel insight into the mechanism underlying EGLN3 inhibition of NFκB signaling and sheds light on the regulation of RIP1 ubiquitination.


Divergent erythroid megakaryocyte fates in Blvrb-deficient mice establish non-overlapping cytoprotective functions during stress hematopoiesis.
Cytoprotective mechanisms of heme oxygenases function by derivatizing heme to generate carbon monoxide, ferrous iron, and isomeric biliverdins, followed by rapid NAD(P)H-dependent biliverdin reduction to the antioxidant bilirubin using two non-overlapping biliverdin reductases that display biliverdin isomer-restricted redox activity. Although cytoprotective functions of heme oxygenases are widely recognized, concomitant effects of downstream biliverdin reductases remain incomplete. A computational model predicated on murine hematopoietic single-cell transcriptomic data identified Blvrb as a biological driver linked to the tumor necrosis factor stress pathway as a predominant source of variation defining hematopoietic cell heterogeneity. In vivo studies using Blvrb-deficient mice established the dispensable role of Blvrb in steady-state hematopoiesis, although model validation using aged Blvrb-deficient mice established an important cytoprotective function in stress hematopoiesis with dichotomous megakaryocyte-biased hematopoietic recovery. Defective stress erythropoiesis was evident in Blvrb-/- spleens and in bone marrow erythroid development, occurring in conjunction with defective lipid peroxidation as a marker of oxidant mishandling. Cell autonomous effects on megakaryocyte lineage bias were documented using multipotential progenitor assays. These data provide the first physiological function of murine Blvrb in a non-redundant pathway of stress cytoprotection. Divergent effects on erythroid/megakaryocyte lineage speciation impute a novel redox-regulated mechanism for lineage partitioning.


A Randomized Phase II Crossover Study of Imatinib or Rituximab for Cutaneous Sclerosis after Hematopoietic Cell Transplantation.
Cutaneous sclerosis occurs in 20% of patients with chronic graft-versus-host disease (GVHD) and can compromise mobility and quality of life. We conducted a prospective, multicenter, randomized, two-arm phase II crossover trial of imatinib (200 mg daily) or rituximab (375 mg/m(2) i.v. weekly × 4 doses, repeatable after 3 months) for treatment of cutaneous sclerosis diagnosed within 18 months (NCT01309997). The primary endpoint was significant clinical response (SCR) at 6 months, defined as quantitative improvement in skin sclerosis or joint range of motion. Treatment success was defined as SCR at 6 months without crossover, recurrent malignancy or death. Secondary endpoints included changes of B-cell profiles in blood (BAFF levels and cellular subsets), patient-reported outcomes, and histopathology between responders and nonresponders with each therapy. SCR was observed in 9 of 35 [26%; 95% confidence interval (CI); 13%-43%] participants randomized to imatinib and 10 of 37 (27%; 95% CI, 14%-44%) randomized to rituximab. Six (17%; 95% CI, 7%-34%) patients in the imatinib arm and 5 (14%; 95% CI, 5%-29%) in the rituximab arm had treatment success. Higher percentages of activated B cells (CD27(+)) were seen at enrollment in rituximab-treated patients who had treatment success (P = 0.01), but not in imatinib-treated patients. These results support the need for more effective therapies for cutaneous sclerosis and suggest that activated B cells define a subgroup of patients with cutaneous sclerosis who are more likely to respond to rituximab.


Dynamic connectedness between FinTech innovation and energy volatility during the war in time of pandemic.
We use an extended joint connectedness technique and the time-varying parameter vector autoregression (ETVP-VAR) method to examine connections between the ARK FinTech Innovation ETF (ARKF), Global X FinTech ETF (FINX), and energy volatility by connectedness as a quality of eight indicators from April 1, 2019, to September 26, 2022. Our results demonstrate that the pattern of ARKF and FINX is picked up as a crucial net shock transmitter that nearly permeates our analyzed sample. Since the COVID-19 epidemic, more people are adopting FinTech partly because of their concern about the disease spreading through social contact and cash handling. Moreover, green bonds are net shock recipients over the long term. Furthermore, during the COVID-19 duration and the Russo-Ukrainian War, shocks transmitted to green bonds soared sharply. By contrast, keeping with the clean energy and crude oil trend, these indicators transmit a network of shocks during the period under study. When considering wind power, it becomes clear that this signal first acts as a net shock transmitter before changing into a net receiver of shocks from mid-2021 onwards. We recognize that the system is a net shock receiver regarding clean power. The dynamics invariably lead the series to change to a net shock transmitter in mid-2021. By mid-2021, the developments always cause the series to transform into a net shock transmitter.


Design, synthesis and structure-activity relationship optimization of phenanthridine derivatives as new anti-vitiligo compounds.
Humans have been suffering from vitiligo for a long time. Target vitiligo drugs have yet been approved. Activation of Wnt/β-catenin signalling has potential in the therapeutic use of vitiligo, so exploring new drugs that specifically directly activate Wnt is worthwhile to obtain new anti-vitiligo agents. In this work, two portions design and synthesis were put into effect. firstly, 17 phenanthridine derivatives with C-4 substitutes were designed and synthesized, which compounds 4, 6, 12, 13 served as H-acceptor with protein showed enhance melanogenesis activity; Secondly, 7 hybrid new scaffolds of compounds were designed and synthesized, scaffold hopping compound 36 that aromatic benzene was replaced pyrazole on ring C showed enhance melanogenesis and tyrosinase activity; The last and most important, a comprehensive optimization and SARs of compound 36 were carried out, compounds 41 and 43 shared phenolic hydroxyl or 3-methyl-pyridine substitutes at C-7 position remarkably improved the capacity of melanogenesis and tyrosinase activity. Compound 43 were identified as new anti-vitiligo agents that specifically activate the Wnt/β-catenin signalling pathway by targeting Axin. Structure-activity relationship analysis implied that H-acceptor substitutions at the C-4 position and phenolic hydroxyl or pyridine substitutions at the C-7 position would improve the activities of the compounds. These findings reveal a new therapeutic strategy for vitiligo, and compounds 41 and 43 may represent potential compounds for vitiligo treatment.


Resistance against Brugia malayi microfilariae induced by a monoclonal antibody which promotes killing by macrophages and recognizes surface antigen(s).
Several monoclonal antibodies were produced following the immunization of mice with infective larvae of Brugia malayi. One of these gives a positive fluorescence reaction on the surface of B. malayi microfilariae and this particular monoclonal antibody (IgM isotype) was able to mediate mouse peritoneal macrophage adherence to, and killing of, B. malayi microfilariae in vitro. Adherence and killing were enhanced by fresh normal mouse serum, suggesting a role for complement. When the same monoclonal antibody was passively transferred to mice harbouring microfilariae in their circulation, a complete clearance of microfilariae was observed in 70% of the animals. This monoclonal antibody was able to recognize antigenic determinants (of 110,000 MW) present on the surface of B. malayi microfilariae by radioimmunoprecipitation.


First molecular detection of Hemolivia and Hepatozoon parasites in reptile-associated ticks on Iriomote Island, Japan.
Hepatozoon and Hemolivia are members of the haemogregarines and are reported in reptiles and reptile-associated ticks. However, no studies have reported on Hepatozoon and Hemolivia in Japanese reptile-associated ticks. This study aimed to molecularly identify and to characterize Hepatozoon and Hemolivia in Japanese reptile-associated ticks, Amblyomma geoemydae (Cantor, 1847) and Amblyomma nitidum (Hirst & Hirst, 1910). A total of 41 and 75 DNA samples from A. geoemydae and A. nitidum ticks, respectively, were used for screening of Hepatozoon and Hemolivia with polymerase chain reaction targeting 18S rDNA. As a result, Hemolivia and Hepatozoon were detected in two A. geoemydae and one A. nitidum, respectively. The sequences of Hemolivia spp. showed a 99.5% (1,050/1,055 bp) identity with Hemolivia parvula (KR069083), and the Hemolivia spp. were located in the same clade as H. parvula in the phylogenetic tree. The sequences of Hepatozoon sp. showed a 98.4% (1,521/1,545 bp) identity with Hepatozoon colubri (MN723844), and the Hepatozoon sp. was distinct from validated Hepatozoon species in the tree. Our findings highlight the first molecular record of Hemolivia in Japan and present the first detection of Hepatozoon in A. nitidum. Further investigations on these tick-borne protozoa are required to understand their life cycle and pathogenicity.


A novel method for detection of exfoliated prostate cancer cells in urine by RNA in situ hybridization.
In the current study, we explore the feasibility of detecting exfoliated prostate cancer cells in urine using an RNA in situ hybridization (RISH) assay. We hypothesized that robust and specific labeling of prostate cancer cells could be achieved in post-digital rectal examination (DRE) urine samples using RISH. We focused on method development, optimization, and analytical evaluation of RISH-based detection of prostate cancer in urine. We optimized a sample collection, processing, and target detection workflow for urine cytology specimens in conjunction with RNA target detection by RISH. We screened a panel of 11 prostate-specific RNA targets, and selected NKX3-1 and PRAC1 as markers for cells of prostate origin and PCA3 as a marker of prostate malignancy. Following analytical validation of a multiplexed NKX3-1/PRAC1/PCA3 assay, we evaluated whether prostate cancer cells can be detected in a pilot cohort of 19 post-DRE specimens obtained from men diagnosed with prostate cancer. Using cytology specimens prepared from spiked urine samples, we established the analytical validity of the RISH assay for detection and visualization of prostate cells in urine. Cells of prostate origin could be readily and specifically identified and separated into benign and malignant cell populations based on the multiplex test that consisted of markers specific for prostate cells (NKX3-1, PRAC1) and prostate cancer cells (PCA3). Upon evaluation of post-DRE urine from a pilot cohort of prostate cancer patients, we identified 11 samples in which prostate cells were present, 6 of which were also positive for prostate cancer cells. Multiplex RISH enables the direct visualization and molecular characterization of individual exfoliated prostate cells in urine. This proof-of-principle study provides evidence supporting the application of RISH as a potential noninvasive tool for prostate cancer detection.


Optimization of 3D-aggregated spheroid model (3D-ASM) for selecting high efficacy drugs.
Various three-dimensional (3D) cell culture methods have been developed to implement tumor models similar to in vivo. However, the conventional 3D cell culture method has limitations such as difficulty in using an extracellular matrix (ECM), low experimental reproducibility, complex 3D cell culture protocol, and difficulty in applying to high array plates such as 96- or 384-plates. Therefore, detailed protocols related to robust 3D-aggregated spheroid model (3D-ASM) production were optimized and proposed. A specially designed wet chamber was used to implement 3D-ASM using the hepatocellular carcinoma (HCC) cell lines, and the conditions were established for the icing step to aggregate the cells in one place and optimized ECM gelation step. Immunofluorescence (IF) staining is mainly used to simultaneously analyze drug efficacy and changes in drug-target biomarkers. By applying the IF staining method to the 3D-ASM model, confocal microscopy imaging and 3D deconvolution image analysis were also successfully performed. Through a comparative study of drug response with conventional 2D-high throughput screening (HTS), the 3D-HTS showed a more comprehensive range of drug efficacy analyses for HCC cell lines and enabled selective drug efficacy analysis for the FDA-approved drug sorafenib. This suggests that increased drug resistance under 3D-HTS conditions does not reduce the analytical discrimination of drug efficacy, also drug efficacy can be analyzed more selectively compared to the conventional 2D-HTS assay. Therefore, the 3D-HTS-based drug efficacy analysis method using an automated 3D-cell spotter/scanner, 384-pillar plate/wet chamber, and the proposed 3D-ASM fabrication protocol is a very suitable platform for analyzing target drug efficacy in HCC cells.


Divergence in inflorescence height: An evolutionary response to pollinator fidelity.
The above-ground heights of inflorescences of 8 species of wild-flowers in a subalpine meadow in the Colorado Rocky Mountains were measured in two successive years. An index of overlap (of height-distributions) was computed for pairwise comparisons of all species. The species were assorted into 4 groups based on their usual pollinators: long-tongued bumblebees (3 species were pollinated by long-tongued bumblebees), short-tongued bumblebees (3 species), hummingbirds (1 species), and solitary bees (1 species). The values of the sample of overlap indices for plants pollinated by the same animals was significantly smaller than the values for plants pollinated by different animals; plants which share pollinators are less alike in height than those that don't share pollinators. It is suggested that this is a result of selection for enhancement of pollinator fidelity. The selective mechanisms, based on the 'horizontal' flight pattern of pollinators and the consequences to the plant of interspecific flights, are discussed.


1987 H. P. Smith award lecture. Eyes for the epidemiologist: the pathologist's role in shaping our understanding of the asbestos-associated diseases.
Asbestosis was first recognized as an entity at autopsy by a pathologist in 1900. Pathologists also discovered the unique relationship of mesothelioma and bronchogenic carcinoma with exposure to asbestos. The observations of preceding pathologists have provided insights that have served as the basis for epidemiologic studies associating asbestos with disease in humans.


Phosphorylation of acidic ribosomal proteins from rabbit reticulocytes by a ribosome-associated casein kinase.
Two acidic proteins from 80-S ribosomes were isolated and purified to homogeneity. The purified acidic proteins could be phosphorylated by casein kinase using [gamma-32P]ATP and [gamma-32P]GTP as a phosphoryl donor. The proteins became phosphorylated in situ, too. Sodium dodecyl sulfate polyacrylamide gel analysis of the purified acidic proteins and 80-S particles showed identical phosphoproteins in the 16 000 dalton region.


Membrane lipid fluidity and its effect on the activation energy of membrane-associated enzymes.
1. The fatty acid composition of mitochondrial membranes from sheep and rats was altered by feeding these animals diets which were rich in unsaturated fatty acids. Changes in membrane lipid fluidity resulting from the altered membrane lipid composition were assessed by determining the upper temperature limit of the disorder-order transition (Tf) and the Arrhenius activation energy (Ea) of succinate oxidase. 2. After feeding the unsaturated fatty acid-rich diet to sheep the Ea, in the temperature range above Tf, increased from 8 to 63 kJ . mol-1 while Tf decreased from 32 to 15 degrees C. Rats fed an unsaturated fatty acid-rich diet exhibited an increase in Ea from 17 to 63 kJ . mol-1 and a decrease in Tf from 23 to 4 degrees C. 3. This decrease in Tf was related to an increase in the ratio of linoleic acid to stearic acid in the membrane lipid. Tf was not related to the proportion of unsaturated fatty acids in the membrane lipids, although an increase in unsaturation usually led to a decrease in Tf. 4. The results show that membrane lipid fluidity has a direct influence on the conformation of the active site of some membrane-associated enzymes, with the result that such enzymes display a higher Ea when the membrane lipids are comparatively more fluid. The increase in Ea of membrane-associated enzymes which accompanies changes in the physical state of membrane suggests that some proteins may phase separate with the more fluid lipids at low temperatures.


Temporal differences between load and movement signal integration in the sensorimotor network of an insect leg.
Nervous systems face a torrent of sensory inputs, including proprioceptive feedback. Signal integration depends on spatially and temporally coinciding signals. It is unclear how relative time delays affect multimodal signal integration from spatially distant sense organs. We measured transmission times and latencies along all processing stages of sensorimotor pathways in the stick insect leg muscle control system, using intra- and extracellular recordings. Transmission times of signals from load-sensing tibial and trochanterofemoral campaniform sensilla (tiCS, tr/fCS) to the premotor network were longer than from the movement-sensing femoral chordotonal organ (fCO). We characterized connectivity patterns from tiCS, tr/fCS, and fCO afferents to identified premotor nonspiking interneurons (NSIs) and motor neurons (MNs) by distinguishing short- and long-latency responses to sensory stimuli. Functional NSI connectivity depended on sensory context. The timeline of multisensory integration in the NSI network showed an early phase of movement signal processing and a delayed phase of load signal integration. The temporal delay of load signals relative to movement feedback persisted into MN activity and muscle force development. We demonstrate differential delays in the processing of two distinct sensory modalities generated by the sensorimotor network and affecting motor output. The reported temporal differences in sensory processing and signal integration improve our understanding of sensory network computation and function in motor control.NEW & NOTEWORTHY Networks integrating multisensory input face the challenge of not only spatial but also temporal integration. In the local network controlling insect leg movements, proprioceptive signal delays differ between sensory modalities. Specifically, signal transmission times to and neuronal connectivity within the sensorimotor network lead to delayed information about leg loading relative to movement signals. Temporal delays persist up to the level of the motor output, demonstrating its relevance for motor control.


Alignment-free similarity analysis for protein sequences based on fuzzy integral.
Sequence comparison is an essential part of modern molecular biology research. In this study, we estimated the parameters of Markov chain by considering the frequencies of occurrence of the all possible amino acid pairs from each alignment-free protein sequence. These estimated Markov chain parameters were used to calculate similarity between two protein sequences based on a fuzzy integral algorithm. For validation, our result was compared with both alignment-based (ClustalW) and alignment-free methods on six benchmark datasets. The results indicate that our developed algorithm has a better clustering performance for protein sequence comparison.


Combined two-photon luminescence microscopy and OCT for macrophage detection in the hypercholesterolemic rabbit aorta using plasmonic gold nanorose.
The macrophage is an important early cellular marker related to risk of future rupture of atherosclerotic plaques. Two-channel two-photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta-based macrophages using plasmonic gold nanorose as an imaging contrast agent. Nanorose uptake by macrophages was identified by TPL microscopy in macrophage cell culture. Ex vivo aorta segments (8 × 8 × 2 mm(3) ) rich in macrophages from a rabbit model of aorta inflammation were imaged by TPL microscopy in combination with OCT. Aorta histological sections (5 µm in thickness) were also imaged by TPL microscopy. Merged two-channel TPL images showed the lateral and depth distribution of nanorose-loaded macrophages (confirmed by RAM-11 stain) and other aorta components (e.g., elastin fiber and lipid droplet), suggesting that nanorose-loaded macrophages are diffusively distributed and mostly detected superficially within 20 µm from the luminal surface of the aorta. Moreover, OCT images depicted detailed surface structure of the diseased aorta. Results suggest that TPL microscopy combined with OCT can simultaneously reveal macrophage distribution with respect to aorta surface structure, which has the potential to detect vulnerable plaques and monitor plaque-based macrophages overtime during cardiovascular interventions.


Dynamic changes of PIRH2 and p27kip1 expression in injured rat sciatic nerve.
p53-induced ring-H2 protein (PIRH2), a newly identified E3 ubiquitin ligase, has been reported to be interacted with p27Kip1 and promote ubiquitination of p27Kip1 independently of p53. p27kip1, a member of the Cip/Kip family of cyclin-dependent kinases inhibitors (CKIs), was shown to control cell cycle progression and promote cell proliferation. While the distribution and function of PIRH2 and p27kip1 in nervous system lesion and regeneration remains unclear. Here, we performed a sciatic nerve injury model in adult rats and studied the dynamic changes of PIRH2 and p27kip1 expression by western blot and RT-PCR in injured rat sciatic nerve. Sciatic nerve crush resulted in a significant up-regulation of PIRH2 and a down-regulation of p27kip1. Besides, we observed that they were expressed widely in both Schwann cells and axons in adult rat sciatic nerve by double immunofluorescence staining. Results obtained by coimmunoprecipitation and double labeling further showed their interaction in the regenerating process. Thus, these results indicate that PIRH2 and p27kip1 likely play an important role in peripheral nerve injury and regeneration.


Experimental study of the topological flow transformations in an aerial vortex bioreactor with a floating washer.
Research into the flow structure in an aerial vortex bioreactor is relevant for developing the methods for growing cell cultures. Determining the optimal cultivation condition for a certain process is especially important in the case when such parameters of the medium as density and viscosity significantly change with the culture growth in the bioreactor. The research of the flow dynamic was carried out in an 8.5 L universal aerial vortex bioreactor, with a washer freely floating on its surface and stabilizing the motion of the working fluid. The regularities of the vortex motion of the culture medium have been determined by Particle Image Velocimetry depending on its volume and the intensity of rotation of the activator, generating vortex motion in the air. The observed vortex structure and its dynamics at increasing flow swirl intensity are established to coincide with the structure of a confined vortex flow in a cylindrical container with no washer for both single and two-fluid configurations. This novel methodology of the flow optimization shows that an ascending swirling jet forms in the vicinity of the bioreactor axis, and a bubble-like vortex breakdown forms in the axial region.


Dynamic regulation of Smad expression during mesenchyme to epithelium transition in the metanephric kidney.
Genetic studies have revealed a role for the transforming growth factor beta (TGFbeta) superfamily growth factors in development of the mouse kidney. In this study, we have characterized developmental expression of Smad proteins, the downstream effectors of TGFbeta superfamily ligands. Immunohistochemistry detecting Smads 1-5 and 8 was performed on 11.25-15.5 dpc kidney sections. We find that Smad proteins are readily detected in metanephric mesenchyme but are coordinately down-regulated in condensing mesenchyme and derivative structures such as renal vesicles. In order to define the stage at which down-regulation occurs, we used the heterologous inducer lithium chloride to promote condensation in isolated cultured mesenchymal explants and subsequently assayed expression of Smad 4. Down-regulation is complete in lithium chloride induced mesenchymal condensates, indicating that Smad regulation takes place at an early stage, prior to mesenchyme to epithelium transition.


Rap G protein signal in normal and disordered lymphohematopoiesis.
Rap proteins (Rap1, Rap2a, b, c) are small molecular weight GTPases of the Ras family. Rap G proteins mediate diverse cellular events such as cell adhesion, proliferation, and gene activation through various signaling pathways. Activation of Rap signal is regulated tightly by several specific regulatory proteins including guanine nucleotide exchange factors and GTPase-activating proteins. Beyond cell biological studies, increasing attempts have been made in the past decade to define the roles of Rap signal in specific functions of normal tissue systems as well as in cancer. In the immune and hematopoietic systems, Rap signal plays crucial roles in the development and function of essentially all lineages of lymphocytes and hematopoietic cells, and importantly, deregulated Rap signal may lead to unique pathological conditions depending on the affected cell types, including various types of leukemia and autoimmunity. The phenotypical studies have unveiled novel, even unexpected functional aspects of Rap signal in cells from a variety of tissues, providing potentially important clues for controlling human diseases, including malignancy.


Comparison of a novel mini-oral sulfate tablet and the conventional oral sulfate tablet in bowel preparation for colonoscopy: a prospective, randomized, investigator-blinded, multicenter, non-inferior, phase 3 trial.
The use of conventional oral sulfate tablets (OSTs) has gained popularity; nonetheless, they may be not only inconvenient to swallow but also difficult to dissolve. A novel mini-OST has recently been developed to enhance compliance with conventional OST use. This study aimed to compare the efficacy, tolerability, and safety between mini-OST and conventional OST. This was a prospective, randomized, investigator-blinded, multicenter, and non-inferior phase 3 trial conducted between September 2022 and December 2022. The efficacy, safety, and tolerability were compared between mini-OST and conventional OST. Exactly 83 patients were evaluated based on a full analysis set (FAS), whereas 82 patients were evaluated as a per-protocol set (PPS). With respect to the efficacy of preparation, successful and high-quality preparation was excellent in the mini-OST and conventional OST groups on both FAS and PPS analyses, without significant differences between the two groups. On the FAS analysis, the satisfaction and tolerability scores were high in both groups, without significant differences. The first bowel movement after taking the investigational product occurred 30 min earlier in the mini-OST group than in the OST group. Mild, moderate, and severe adverse events (AEs) were comparable between the two groups; however, any AEs were more common in the mini-OST group than in the conventional OST group overall. Compared with conventional OST, the novel mini-OST preparation showed similar efficacy, tolerability, and safety, including mild eight solicited AEs and moderate-to-severe AEs. Clinical trial registration NCT05670470.


A role for metallothionein in the pathogenesis of diabetes and its cardiovascular complications.
It has been suspected for a long time that zinc has a role in various aspects of diabetes, but specific molecular targets of zinc remained largely elusive. Recent discoveries of associations between diabetes and polymorphisms in human genes now suggest that proteins that control the cellular availability of zinc ions are involved. One protein is the zinc transporter ZnT-8 that supplies pancreatic beta-cells with zinc. The other is metallothionein 1A, a member of a protein family that links zinc and redox metabolism. Changes in the availability of zinc ions modulate insulin signaling and redox processes. Both zinc and metallothionein protect cells against the redox stress that occurs in diabetes and contributes to its progression towards diabetic complications, including heart disease.


Chronic inflammation does not appear to modify the homozygous hereditary hemochromatosis phenotype.
The mechanism of excessive iron storage in patients with hereditary hemochromatosis caused by mutations of the HFE gene seems to be a failure to up-regulate hepcidin in the face of increased body iron. Since the cytokines IL-1 and IL-6 stimulate hepcidin transcription in the absence of HFE, chronic inflammatory states might counteract the effect of HFE mutations. We measured the pre-phlebotomy plasma levels of C reactive protein (CRP) and of interleukin 6 (IL-6) in homozygotes for the C282Y mutation of HFE. There was no difference in these levels in subjects with high iron stores than in those with low iron stores, suggesting that the phenotypic differences between such homozygotes is not appreciably affected by ongoing chronic inflammation.


Amelioration of diet-induced nonalcoholic steatohepatitis in rats by Mn-salen complexes via reduction of oxidative stress.
Nonalcoholic steatohepatitis (NASH), a progressive stage of nonalcoholic fatty liver disease (NAFLD), is characterized by steatosis (accumulation of triacylglycerols within hepatocytes) along with inflammation and ballooning degeneration. It has been suggested that oxidative stress may play an important role in the progress of NAFLD to NASH. The aim of present study was to determine whether antioxidant supplementations using EUK-8, EUK-134 and vitamin C could improve the biochemical and histological abnormalities associated with diet-induced NASH in rats. NASH was induced in male N-Mary rats by feeding a methionine - choline deficient (MCD) diet. The rats were fed either normal chow or MCD diet for 10 weeks. After NASH development, the MCD-fed rats were randomly divided into four groups of six: the NASH group that received MCD diet, the EUK-8 group which was fed MCD diet plus EUK-8, the EUK-134 group which was fed MCD diet plus EUK-134 and the vitamin C group which received MCD diet plus vitamin C. EUK-8, EUK-134 and vitamin C (30 mg/kg body weight/day) were administered by gavage for eight weeks. Treatment of MCD-fed rats with salens reduced the sera aminotransferases, cholesterol, low density lipoprotein contents, the extent of lipid peroxidation and protein carbonylation whereas the HDL-C cholesterol levels were significantly increased. In addition, EUK-8 and EUK-134 improved steatosis, ballooning degeneration and inflammation in liver of MCD-fed rats. Antioxidant (EUK-8, EUK-134 and vitamin C) supplementation reduces NASH-induced biochemical and histological abnormalities, pointing out that antioxidant strategy could be beneficial in treatment of NASH.


Higher versus lower oxygenation targets in COVID-19 patients with severe hypoxaemia (HOT-COVID) trial: Protocol for a secondary Bayesian analysis.
Respiratory failure is the main cause of mortality and morbidity among ICU patients with coronavirus disease 2019 (COVID-19). In these patients, supplemental oxygen therapy is essential, but there is limited evidence the optimal target. To address this, the ongoing handling oxygenation targets in COVID-19 (HOT-COVID) trial was initiated to investigate the effect of a lower oxygenation target (partial pressure of arterial oxygen (PaO2 ) of 8 kPa) versus a higher oxygenation target (PaO2 of 12 kPa) in the ICU on clinical outcome in patients with COVID-19 and hypoxaemia. The HOT-COVID is planned to enrol 780 patients. This paper presents the protocol and statistical analysis plan for the conduct of a secondary Bayesian analysis of the primary outcome of HOT-COVID being days alive without life-support at 90 days and the secondary outcome 90-day all-cause mortality. Furthermore, both outcomes will be investigated for the presence heterogeneity of treatment effects based on four baseline parameters being sequential organ failure assessment score, PaO2 /fraction of inspired oxygen ratio, highest dose of norepinephrine during the 24 h before randomisation, and plasma concentration of lactate at randomisation. The results of this pre-planned secondary Bayesian analysis will complement the primary frequentist analysis of the HOT-COVID trial and may facilitate a more nuanced interpretation of the trial results.


Seasonal Variation of Mercury and Its Isotopes in Atmospheric Particles at the Coastal Zhongshan Station, Eastern Antarctica.
Mercury (Hg) is a globally spread trace metal due to its long atmospheric residence time. Yet, our understanding of atmospheric processes (e.g., redox reactions and deposition) driving Hg cycling is still limited, especially in polar regions. The Antarctic continent, by virtue of its remoteness, is the perfect location to investigate Hg atmospheric processes in the absence of significant local anthropogenic impact. Here, we present the first 2 year record (2016-2017) of total suspended particulate mercury (PHg) concentrations along with a year-round determination of an Hg stable isotopic composition in particles collected at Zhongshan Station (ZSS), eastern Antarctic coast. The mean PHg concentration is 21.8 ± 32.1 pg/m3, ranging from 0.9 to 195.6 pg/m3, and peaks in spring and summer. The negative mass-independent fractionation of odd Hg isotopes (odd-MIF, average -0.38 ± 0.12‰ for Δ199Hg) and the slope of Δ199Hg/Δ201Hg with 0.91 ± 0.12 suggest that the springtime isotope variation of PHg is likely caused by in situ photo-oxidation and reduction reactions. On the other hand, the increase of PHg concentrations and the observed odd-MIF values in summer are attributed to the transport by katabatic winds of divalent species derived from the oxidation of elemental Hg in the inland Antarctic Plateau.


3' processing of tRNA precursors in ribonuclease-deficient Escherichia coli. Development and characterization of an in vitro processing system and evidence for a phosphate requirement.
In order to determine the mechanism and enzyme(s) responsible for 3' processing of tRNA precursors, we have developed an in vitro processing system that uses as substrates two SP6 RNA polymerase-generated transcripts of the gene for tRNA(Tyrsu3)+ that contain 49 extra 5'-nucleotides and either 5 or 25 extra 3'-nucleotides. A high speed supernatant fraction from an Escherichia coli strain deficient in five ribonucleases was found to accurately process both tRNA precursors in this system to the size of mature tRNA(Tyr). Final 3' end processing of each precursor occurs in an exonucleolytic manner to generate the correct 3' terminus; however, a prior endonucleolytic cleavage also is observed in processing of the longer precursor. The system requires Mg2+ and works optimally at about 50 mM KCl and pH 8-9. Dialysis of the supernatant fraction leads to loss of processing activity but can be restored to normal by the addition of inorganic phosphate or arsenate. Furthermore, nucleoside diphosphates are a product of the processing reaction. These data indicate that 3' processing in RNase-deficient extracts involves a phosphorolytic reaction. On the other hand, phosphate is not required for processing in RNase+ extracts, although it does aid in processing of the longer precursor. The usefulness of this in vitro system for studies of tRNA processing and the identity of the phosphate-requiring enzyme are discussed.


Employment of electrochemically synthesized TGA-CdSe quantum dots for Cr(3+) determination in vitamin supplements.
The fluorescence quenching of TGA-CdSe quantum dots (QDs) was used for Cr(3+) quantification in vitamin supplements. The QD was electrochemically synthesized, demonstrating high reproducibility with control of particle size, thus making it a clean method, without the presence of reducing agents. Under ideal conditions, with the fluorescence band at 551 nm (excitation 365 nm), the maximum fluorescence quenching was observed at pH 4.0, with a time of 200 s for each data acquisition. Under optimum experimental conditions, linear quenching was observed for Cr(3+) in the range of 25.0-325.0 ng L(-1) (R=0.9996, n=6), a limit of detection of 5.67 ng L(-1), and relative standard deviation of 4.43% (n=10). The recovery test for Cr(3+) quantification in vitamin supplements presented results from 82% to 98%. These Cr(3+) determination results were compared to the same vitamin supplement sample using flame atomic absortion spectrometry (FAAS) method, and no significant differences were observed at 95% confidence level.


Prenatal screening for cystic fibrosis: attitudes and responses of participants.
A screening programme to detect cystic fibrosis heterozygotes has been running in the antenatal clinics of a major Edinburgh maternity hospital for more than 2 years. A questionnaire was used to assess participants' knowledge of the genetics of the disorder and their attitudes to being screened. The respondents were 64 female heterozygotes and 63 of their non-heterozygous male partners, 101 female controls and 100 male controls. Although the two groups of controls received far less direct information than the carriers and their partners, all four groups were well informed about the genetics of cystic fibrosis and the significance of being a gene carrier. A majority of each group felt that adequate information had been given in the information leaflet, that they understood the purpose of screening and that they were glad to have participated. There was a consensus that CF carrier testing should be routinely offered to pregnant women, and also that it should be available in family planning clinics and GP health centres, but not in schools.


Computed tomography of the orbit - A review and an update.
Computed tomography (CT) and magnetic resonance imaging (MRI) of the orbit have been competing for the hearts and minds of health care providers for well over 2 decades. While several drawbacks pertaining to CT have been outlined since the introduction of MRI, CT remains the standard diagnostic test for evaluating cross-sectional, 2 or 3-dimensional images of the body.


Molecular characterization of flavanone 3-hydroxylase gene and flavonoid accumulation in two chemotyped safflower lines in response to methyl jasmonate stimulation.
Among secondary metabolites, flavonoids are particularly crucial for plant growth, development, and reproduction, as well as beneficial for maintenance of human health. As a flowering plant, safflower has synthesized a striking variety of flavonoids with various pharmacologic properties. However, far less research has been carried out on the genes involved in the biosynthetic pathways that generate these amazing flavonoids, especially characterized quinochalcones. In this study, we first cloned and investigated the participation of a presumed flavanone 3-hydroxylase gene (F3H) from safflower (CtF3H) in a flavonoid biosynthetic pathway. Bioinformation analysis showed that CtF3H shared high conserved residues and confidence with F3H from other plants. Subcellular localization uncovered the nuclear and cytosol localization of CtF3H in onion epidermal cells. The functional expressions of CtF3H in Escherichia coli BL21(DE3)pLysS cells in the pMAL-C5x vector led to the production of dihydrokaempferol when naringenin was the substrate. Furthermore, the transcriptome expression of CtF3H showed a diametrically opposed expression pattern in a quinochalcone-type safflower line (with orange-yellow flowers) and a flavonol-type safflower line (with white flowers) under external stimulation by methyl jasmonate (MeJA), which has been identified as an elicitor of flavonoid metabolites. Further metabolite analysis showed the increasing tendency of quinochalcones and flavonols, such as hydroxysafflor yellow A, kaempferol-3-O-β-D-glucoside, kaempferol-3-O-β-rutinoside, rutin, carthamin, and luteolin, in the quinochalcone-type safflower line. Also, the accumulation of kaempferol-3-O-β-rutinoside and kaempferol-3-O-β-D-glucoside in flavonols-typed safflower line showed enhanced accumulation pattern after MeJA treatment. However, other flavonols, such as kaempferol, dihydrokaempferol and quercetin-3-O-β-D-glucoside, in flavonols-typed safflower line presented down accumulation respond to MeJA stimulus. Our results showed that the high expression of CtF3H in quinochalcone-type safflower line was associated with the accumulation of both quinochalcones and flavonols, whereas its low expression did not affect the increased accumulation of glycosylated derivatives (kaempferol-3-O-β-rutinoside and rutin) in flavonols-typed safflower line but affect the upstream precursors (D-phenylalanine, dihydrokaempferol, kaempferol), which partly revealed the function of CtF3H in different phenotypes and chemotypes of safflower lines.


Functional dissection of an early Drosophila chorion gene promoter: expression throughout the follicular epithelium is under spatially composite regulation.
We have fused various DNA sequences located upstream of the Drosophila melanogaster s36 chorion gene TATA box to a heterologous basal promoter and reporter gene (hsp70/lacZ). The expression of these constructs, following P-element-mediated germline transformation, was examined in 144 independent lines by histological staining of dissected ovaries for beta-galactosidase activity. A short 84 bp segment of the proximal 5' flanking DNA was sufficient to confer a wild-type gene expression pattern, including temporal specificity for early choriogenic follicles. Surprisingly, initial expression was very localized at the anterior and posterior poles of the follicle. The downstream half of that DNA segment permitted expression at both poles, but especially at the anterior tip, while the upstream half only favored expression in the posterior pole; these results suggested the existence of multiple, spatially specific cis-regulatory elements. When the proximal 84 bp segment was placed 1.5 kb upstream of the basal promoter, beta-galactosidase activity was observed in an altered spatial pattern, indicating that the cis-regulatory element(s) that favor expression in the posterior half of the follicle are position independent, while the element(s) that favor expression elsewhere in the follicle are position sensitive. A distal regulatory segment containing redundant DNA element(s) specific for expression in the anterior pole was identified much further upstream of s36. Thus, the expression of this chorion gene throughout the follicular epithelium is actually composite, occurring in distinct spatial domains under the control of corresponding DNA elements.


WITHDRAWN: NMDA receptors are required in memory formation in Drosophila mushroom body.
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The morbidity of carotid endarterectomy.
The results of 72 carotid endarterectomy operations carried out by three consultant surgeons were analysed. Thirty-eight operations (32.8 per cent) were performed in patients with mild or resolving stroke and 33 operations (45.9 per cent) in patients with transient ischaemic attacks and/or amaurosis fugax. Five patients (7 per cent) died in the early postoperative period, 2 as a result of strokes and the others from cardiac causes. Seventy-six per cent of patients had a satisfactory long term result, remaining free from storke, blindness or recurrent transient ischaemic attacks over the follow-up period of 1--7 years. There were 10 patients (13.8 per cent), including 2 who died, who developed early postoperative neurological deficits; 4 (5.5 per cent) had transient defects and 6 (8.3 per cent) had permanent deficits. Of these patients, 4 had proved occlusions of the reconstructed artery. In 3 patients, cerebral embolization during operation was suspected as the cause. There were, in addition, 2 asymptomatic patients whose reconstruction was shown to be occluded on follow-up postoperative angiography. The continued absence of symptoms, therefore, is no guarantee that the reconstruction is patent and some more objective method of assessment is necessary if the results of carotid endarterectomy are to be critically analysed. The incidence of neurological deficits could, it is believed, be reduced by greater experience with the operation, the routine use of peroperative intra-arterial shunting and peroperative angiography to confirm the patency of the reconstruction.


Long non-coding RNA HOXA11-AS contributes to the formation of keloid by relieving the inhibition of miR-182-5p on ZNF217.
Long non-coding RNA (lncRNA) dysregulation is demonstrated to be associated with disease progression. Mounting studies show that lncRNA promotes or inhibits the development of keloid. We aimed to disclose the role of homebox A11 antisense RNA (HOXA11-AS) in the formation of keloid. Quantitative real-time PCR (qPCR) was adopted for expression analysis of HOXA11-AS, miR-182-5p and zinc finger protein 217 (ZNF217) mRNA, and the expression of ZNF protein and marker proteins was detected by western blot. Cell proliferation, cell migration and cell apoptosis were investigated using CCK-8 assay, wound healing assay and flow cytometry assay, respectively. The potential interplay between miR-182-5p and HOXA11-AS or ZNF217 was verified by dual-luciferase reporter assay, RIP assay and pull-down assay. The role of HOXA11 in vivo was studied by establishing animal models. HOXA11-AS was highly expressed in tissues and fibroblasts of keloid. Deficiency of HOXA11-AS blocked the proliferation and migration of keloid fibroblasts and induced fibroblast apoptosis. HOXA11-AS directly combined to miR-182-5p whose downregulation reversed the effects of HOXA11-AS knockdown. ZNF217 was a target of miR-182-5p, and HOXA11-AS indirectly promoted ZNF217 expression by binding to miR-182-5p. MiR-182-5p enrichment also blocked keloid fibroblast proliferation, survival and migration, while further ZNF217 overexpression abolished these effects. HOXA11-AS knockdown also hindered the growth of keloid in mouse models. High expression of HOXA11-AS promoted the formation and growth of keloid through the upregulation of ZNF217 by targeting miR-182-5p, and the inhibition of HOXA11-AS might be a novel strategy to prevent keloid development.


Quantitative studies on the properdin-complement system. II. Kinetics of the reaction between properdin and zymosan.
The reaction of human, bovine, and pig properdin with excess zymosan exhibits an induction period followed by a rapidly accelerating formation of the properdin-zymosan (PZ) complex. Two methods for determining the quantity of PZ complex are given, one using human serum lacking properdin, the other using a fraction of pig serum containing complement components, C'1, C'3 and C'4.


Detoxification of hexavalent chromium by Leucobacter sp. uses a reductase with specificity for dihydrolipoamide.
Leucobacter sp. belongs to the metal stressed community and possesses higher tolerance to metals including chromium and can detoxify toxic hexavalent chromium by reduction to less toxic trivalent chromium. But, the mechanism of reduction of hexavalent chromium by Leucobacter sp. has not been studied. Understanding the enzyme catalyzing reduction of chromium is important to improve the species for application in bioremediation. Hence, a soluble reductase catalyzing the reduction of hexavalent chromium was purified from a Leucobacter sp. and characterized. The pure chromate reductase was obtained from the cell-free extract through hydrophobic interaction and gel filtration column chromatographic methods. It was a monomeric enzyme and showed similar molecular weights in both gel filtration (∼68 KDa) and SDS-PAGE (64 KDa). It reduced Cr(VI) using both NADH and NADPH as the electron donor, but exhibited higher activity with NADH. The optimal activity was found at pH 5.5 and 30 °C. The K(m) and V(max) for Cr(VI) reduction with NADH were 46.57 μM and 0.37 μmol min(-1) (mg protein) (-1), respectively. The activity was inhibited by p-hydroxy mercury benzoate, Ag(2+) and Hg(2+) indicating the role of thiol groups in the catalysis. The spectrophotometric analysis of the purified enzyme showed the absence of bound flavin in the enzyme. The N-terminal amino acid sequence and LC/MS analysis of trypsin digested purified enzyme showed similarity to dihydrolipoyl dehydrogenase. The purified enzyme had dihydrolipoyl dehydrogenase activity with dihydrolipoamide as the substrate, which suggested that Leucobacter sp. uses reductase with multiple substrate specificity for reduction of Cr(VI) detoxification.


Assessing Use, Usefulness, and Application of the High Impact Practices in Family Planning Briefs and Strategic Planning Guides.
In global health, persistent barriers and challenges to bridging the gap between research and practice remain critical to address in most health areas. The High Impact Practices (HIPs) briefs and strategic planning guides aim to bridge the know-do gap in family planning (FP) by facilitating research utilization and knowledge sharing and also providing a summary of experiential knowledge from experts. The purpose of this qualitative study was to assess the use, usefulness, and application of these 2 knowledge products developed by the HIP Partnership for decision-makers and implementers in low- and middle-income countries (LMICs). This research used in-depth interviews with FP stakeholders from various LMICs to assess the use, usefulness, and application of 2 HIP products for FP decision-makers and implementers. The analysis was shaped by an adapted logic model framework to assess HIP product reach, engagement, usefulness, learning, and action. We interviewed 35 FP professionals from January to March 2021. Participants reported that HIP products have a wide reach, have garnered positive engagement, and were useful. Participants generally liked the current format of the HIP products and reported using them to inform program design, guide discussions with partners, enhance personal knowledge, support advocacy work, and strengthen the guidance they provide to colleagues in the field. The participants shared important feedback to improve the development and dissemination of HIP products, particularly a need to enhance local access and use. The study highlighted the importance of knowledge products, such as the HIP briefs and strategic planning guides, to make evidence and experiential knowledge accessible to a wide audience. These can be valuable tools for policymakers and program implementers to ensure public health practices are evidence-based and integrate experiential knowledge.


Modulation of apoptosis in the mouse brain after morphine treatments and morphine withdrawal.
We have examined the effects of acute or chronic morphine and naltrexone-precipitated withdrawal on mouse brain apoptotic cell death. The associated changes in the expression of apoptosis regulatory proteins were also analyzed. After a single dose of morphine, no apoptotic cells were detected by TUNEL or active caspase-3 immunocytochemistry. Concurrently, a down-regulation of the proapoptotic proteins FasL and Bad was detected in cortical lysates. On the other hand, the brains of chronic-morphine-treated mice and abstinent mice exhibited scattered apoptotic neurons and astrocytes throughout the brain. This neurotoxic effect was accompanied by up-regulation of the proapoptotic proteins FasL, Fas, and Bad and the active fragments of caspases-8 and -3 in cortical and hippocampal lysates. Abstinent mice also displayed a reduced expression of the antiapoptotic protein Bcl-2. No changes on t-Bid expression were detected under any experimental condition. These results suggest a neurotoxic effect exerted by chronic, but not acute, morphine and its withdrawal by activating both the intrinsic and the extrinsic apoptotic pathways. The possible clinical implications of our findings are discussed.


Cell-Type-Specific Immune Dysregulation in Severely Ill COVID-19 Patients.
Recent studies have demonstrated immunologic dysfunction in severely ill coronavirus disease 2019 (COVID-19) patients. We use single-cell RNA sequencing (scRNA-seq) to analyze the transcriptome of peripheral blood mononuclear cells (PBMCs) from healthy (n = 3) and COVID-19 patients with moderate disease (n = 5), acute respiratory distress syndrome (ARDS, n = 6), or recovering from ARDS (n = 6). Our data reveal transcriptomic profiles indicative of defective antigen presentation and interferon (IFN) responsiveness in monocytes from ARDS patients, which contrasts with higher responsiveness to IFN signaling in lymphocytes. Furthermore, genes involved in cytotoxic activity are suppressed in both natural killer (NK) and CD8 T lymphocytes, and B cell activation is deficient, which is consistent with delayed viral clearance in severely ill COVID-19 patients. Our study demonstrates that COVID-19 patients with ARDS have a state of immune imbalance in which dysregulation of both innate and adaptive immune responses may be contributing to a more severe disease course.


Hair Biology: Growth and Pigmentation.
Healthy hair is vital to identity. Understanding the intricate anatomy and physiology of hair provides insight into the aging process and the eventual loss of either hair pigmentation or hair shafts. Several biologics are available that have enabled altering or slowing the aging process of hair, but, unfortunately, no agent exists that can reverse the natural course. The commonly used biologics are discussed.


29-Deoxymaklamicin, a new maklamicin analogue produced by a genetically engineered strain of Micromonospora sp. NBRC 110955.
Maklamicin is a spirotetronate-class antibiotic produced by Micromonospora sp. NBRC 110955, and a polyketide assembly line and a glycerate utilization system are involved in its biosynthesis. One tailoring step in the biosynthesis is predicted to be post-polyketide synthase (PKS) modification, which seems to be catalysed by putative cytochrome P450 monooxygenases, MakC2 and/or MakC3. In this study, we characterized makC2 and makC3 in the biosynthesis of maklamicin and identified a new maklamicin analogue from a makC2 disruptant. Gene deletion of makC2 resulted in the complete loss of maklamicin production with concomitant accumulation of a new compound (29-deoxymaklamicin), while gene deletion of makC3 did not affect the maklamicin production, indicating that 29-deoxymaklamicin is an intermediate in the biosynthetic pathway of maklamicin and should serve as the substrate of MakC2. 29-Deoxymaklamicin showed strong-to-modest anti-microbial activity against gram-positive bacteria. The fact that Streptomyces avermitilis heterologously expressing makC2 successfully converted 29-deoxymaklamicin into maklamicin confirmed that MakC2 is the final-step hydroxylase in the formation of mature maklamicin.


Chronic cerebral hypoperfusion induces striatal alterations due to the transient increase of NO production and the depression of glutathione content.
We examined the effects of chronic cerebral hypoperfusion on the endogenous oxidative stress-related indices, nitrite and nitrate (NOx) concentration, glutathione (GSH) content, superoxide dismutase and catalase activities, and thiobarbituric acid-reactive substances level in the rat striatum, to clarify the participation of oxidative stress in the chronic cerebral hypoperfusion-induced alterations. Our present results indicate that chronic cerebral hypoperfusion produces oxidative stress and disturbs intracellular redox regulation in two distinct phases: at 1 day, "acute" and at 6 weeks, "chronic" alterations after the operation. Therefore, striatal neural cell damage may be mainly attributed to the transient increase of NOx production at 1 day after, and the delayed reduction of muscarinic acetylcholine receptor binding in the striatum may be mostly attributed to the continuous depression of GSH content from the 1st to the 6th postoperative week. In particular, the continuous GSH depression may be considered to accompany the pathophysiology of chronic cerebral hypoperfusion.


Patterns and controls of vegetation productivity and precipitation-use efficiency across Eurasian grasslands.
Elucidating aboveground net primary production (ANPP) and precipitation-use efficiency (PUE) spatial variations and mechanisms are essential for predicting how ecosystem functioning will respond to future climate change. However, a comprehensive recognition of spatial patterns of ANPP and PUE across continental scale is still lacking. Here, we integrated long-term GIMMS NDVI remote sensing, field survey ANPP and meteorological datasets to reveal the spatial variations and controls of ANPP and PUE across Eurasian grasslands for the first time. The results showed that the mean value of ANPP and PUE of Eurasian grasslands were 40.20 ± 0.40 g C m-2 yr-1 and 0.15 ± 0.01 g C m-2 mm-1, respectively. At the continental scale, the ANPP and PUE showed unimodal patterns along mean annual precipitation (MAP) and hydrothermal index (HT) gradients, while a piecewise linear pattern along mean annual temperature (MAT) gradients. The MAP exerted positive effect on the ANPP in desert and temperate grasslands, while negative effect on the ANPP in alpine grasslands. Conversely, the MAT negatively affected the ANPP in desert and temperate grasslands, while positively affected the ANPP in alpine grasslands. The results indicated that the hydrothermal conditions coupled with the transition of vegetation types and its different responses combinedly shaped the spatial patterns of ANPP and PUE in Eurasian grasslands. This study advanced our knowledge of the spatial variations of ANPP and PUE at continental scale, providing theoretical information for predicting productivity and water use changes of arid and semi-arid grasslands under climate change in the future.


Assessment of the utility of routinely collected cattle census and disposal data for syndromic surveillance.
Census and disposal data provide a multipurpose source of information on cattle mortality. The retrospective analyses we conducted on the data gathered in the National Cattle Register produced relevant information for describing and modelling the cattle mortality baseline and evaluating the impact of the 2007-2008 Blue Tongue epidemic on the French cattle population. This work was conducted retrospectively but showed that monitoring cattle mortality near real time could help detecting unexpected events. We are thus currently working on a timely and automated system to monitor cadaver disposal requests received by rendering plants, thanks to a data interchange system recently implemented between the Ministry of Agriculture and the fallen stock companies. Besides technical and methodological challenges, using these data for surveillance purposes raises epidemiological questions that still need to be answered. The question remains notably as to whether an abnormal increased mortality is a sensitive and timely signal for detecting unexpected health events. It appears also very challenging to identify the most adequate surveillance scale (time, space and population) and the most adequate anomaly detection algorithms to apply when the characteristics of the signals to be detected (shape, amplitude, etc.) are not known a priori. In Human health, similar systems have not yet proven their ability to detect unexpected events earlier than classical surveillance systems currently in place, but they have already demonstrated their value for real time assessment of identified and potentially dangerous events. Combined with traditional surveillance systems, we think that monitoring routinely collected data could improve the surveillance of the animal population health. Even if not used for detection purposes, cattle mortality monitoring could be used to rapidly produce information on the impact and evolution of identified events, what would facilitate decision-making regarding management measures and improve the communication.


Robustness of Hill's overlapping-generation method for calculating Ne to extreme patterns of reproductive success.
For species with overlapping generations, the most widely used method to calculate effective population size (Ne) is Hill's, the key parameter for which is lifetime variance in offspring number ([Formula: see text]). Hill's model assumes a stable age structure and constant abundance, and sensitivity to those assumptions has been evaluated previously. Here I evaluate the robustness of Hill's model to extreme patterns of reproductive success, whose effects have not been previously examined: (1) very strong reproductive skew; (2) strong temporal autocorrelations in individual reproductive success; and (3) strong covariance of individual reproduction and survival. Genetic drift (loss of heterozygosity and increase in allele frequency variance) was simulated in age-structured populations using methods that generated no autocorrelations or covariances (Model NoCor); or created strong positive (Model Positive) or strong negative (Model Negative) temporal autocorrelations in reproduction and covariances between reproduction and survival. Compared to Model NoCor, the other models led to greatly elevated or reduced [Formula: see text], and hence greatly reduced or elevated Ne, respectively. A new index is introduced (ρα,α+), which is the correlation between (1) the number of offspring produced by each individual at the age at maturity (α), and (2) the total number of offspring produced during the rest of their lifetimes. Mean ρα,α+ was ≈0 under Model NoCor, strongly positive under Model Positive, and strongly negative under Model Negative. Even under the most extreme reproductive scenarios in Models Positive and Negative, when [Formula: see text] was calculated from the realized population pedigree and used to calculate Ne in Hill's model, the result accurately predicted the rate of genetic drift in simulated populations. These results held for scenarios where age-specific reproductive skew was random (variance ≈ mean) and highly overdispersed (variance up to 20 times the mean). Collectively, these results are good news for researchers as they demonstrate the robustness of Hill's model even in extreme reproductive scenarios.


[Indication, technic and value of arthroscopy of the upper ankle joint].
The authors demonstrate on several joint preparations that essential parts of the upper ankle joint can be directly looked at by endoscopic examination with access at three different points. In case of osteochondrous fractures such as osteochondrosis dissecans, arthroscopy can be a means to determine the therapeutically relevant stages. A series of clinical examples is presented in order to demonstrate that this method, which is still seldom employed, represents a considerable extension of diagnostic and also therapeutic possibilities.


[Therapeutic utilization of stem cells in orthopedics].
Stem cells are becoming increasingly more important in the field of regenerative medicine. Adult mesenchymal stem cells (MSCs) are harvested predominantly from bone marrow or adipose tissue, are already being used in the clinical setting and have a low potential for side effects. In orthopedics, experience has been gained in the treatment of bone defects, non-unions, cartilage defects, osteoarthritis and tendon pathologies. The current data are derived from case studies and randomized controlled trials are missing; therefore, there are many open questions concerning the optimal cell source, number of cells, administration technique (e.g. injections and matrices) or combinations with growth factors; however, it is evident from the data that MSCs have a positive effect on tissue regeneration and are safe to use.


Assessment of knowledge, attitude, and practice on livestock traceability among cattle farmers and cattle traders in peninsular Malaysia and its impact on disease control.
The ability to trace the movement of animals and their related products is key to success in animal disease control. To ensure that a traceability system is optimized, livestock farmers and traders must have good appreciation and understanding about animal tracing. The present study examined the traceability of cattle in Malaysia vis-à-vis the domains of knowledge, attitude, and practice among cattle farmers and traders. A total of 543 farmers and traders in Peninsular Malaysia were interviewed. The results revealed that over 60% of the respondents had satisfactory knowledge and attitude about cattle movement and traceability. A lower proportion of the respondents (49%) were involved in appropriate practice that facilitated traceability of cattle. We found that the type of husbandry system and stakeholders' participation in livestock management-specific short courses were positively associated with satisfactory knowledge, attitude, and practice. A structured education and training program should be formulated to improve these domains so that the benefit of traceability becomes clear, paving the way to a successful traceability program.


Preliminary results of HBV DNA testing of Polish haemophilia patients--lack of occult HBV infection.
Identification of hepatitis B virus (HBV) infection in the absence of surface antigen (HbsAg) became possible with the introduction of HBV DNA detection methods. Such occult HBV infection was diagnosed recently in about half of the Japanese HBsAg-negative haemophilia patients. The aim of our study was to assess the prevalence of occult HBV infection in Polish severe haemophilia population on the sample of 115 haemophilia A and B patients (mean age 34.9 +/- 10.9) treated with non-virus inactivated clotting factor preparations before 1995. HBV DNA was detected in nine HBsAg-positive patients (7.8%). The mean HBV DNA load was 72,800 IU mL(-1) (250-400,000 IU mL(-1)). Hepatitis C virus (HCV) RNA was found in six out of nine HBV-positive patients. In conclusion, HBV DNA was identified only in HBsAg-positive patients. Unlike in Japan, the frequency of occult HBV infection in Polish haemophilia population seems extremely rare or absent.


Debunking Lesbian Bed Death: Using Coarsened Exact Matching to Compare Sexual Practices and Satisfaction of Lesbian and Heterosexual Women.
The current study examined the prevalence and correlates of over 50 sexual practices in a national survey of heterosexual and lesbian women in relationships. Coarsened exact matching was used to create comparable samples of heterosexual (n = 2510) and lesbian (n = 283) women on six demographic factors, including relationship length. Heterosexual and lesbian women were equally likely to be sexually satisfied (66% heterosexual women vs. 68% lesbian women). Compared to heterosexuals, lesbians were more likely to report having sex 0-1 times per month (11% vs. 23%) and were less likely to report having sex greater than once per month (89% vs. 77%). Among women who had been in relationships for longer than 5 years, heterosexual women were less likely than lesbian women to report having sex 0-1 times per month (15%; 42%). This steeper drop in sexual frequency among lesbian women than heterosexual women has pejoratively been labeled lesbian bed death. Rather than accept the label "lesbian bed death" as characterizing these sexual relationships, we turn our attention to what we call lesbian bed intimacies: the myriad ways that lesbian women incorporate behaviors promoting emotional connection, romance, and mood setting, as well as relying on a wide variety of specific sexual acts (e.g., use of sex toys) and sexual communication. Compared to heterosexual women, lesbian women were more likely to usually to always receive oral sex during sex in the past month (28%; 47%) and to use sex toys in the past year (40%; 62%). In their last sexual encounter, lesbian women were more likely to say "I love you" (67%; 80%), have sex longer than 30 min (48%; 72%), and engage in gentle kissing (80%; 92%). These intimacies likely help explain why sexual satisfaction was similar in these groups despite notable differences in sexual frequency.


Identification of radioresistance-associated proteins in human nasopharyngeal carcinoma cell lines by proteomic analysis.
Radiotherapy is the mainstay of treatment modality for human nasopharyngeal carcinoma (NPC), but in some cases, the disease is radioresistant. This study aimed to identify proteins potentially responsible for radioresistance of NPC by a proteomic approach. The radioresistant human NPC cell line CNE-2R and its parental cell line CNE-2 were analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/time-of-flight mass spectrometry analysis. The candidate proteins were validated by Western blot analysis. In total, 16 differentially expressed proteins were identified; among them, Nm23 H1 was significantly increased, while Annexin A3 was significantly downregulated in CNE-2R cells, compared to CNE-2 cells. Our findings provide a platform for further characterization of the proteins implicated in radioresistance and suggest that these proteins could be exploited as a biomarker for predicting the NPC response to radiotherapy in the clinic.


Crash proximity and equivalent property damage calculation techniques: An investigation using a novel horizontal curve dataset.
Despite the numerous breakthroughs in crash analytics, there remains a lack of consensus among safety practitioners as to the optimal method for locating high crash locations. Two critical components in the traffic safety analysis process not agreed upon are 1) how the crash distance to a target location is included in the analysis and 2) how crashes are weighted based on crash-related characteristics. For example, the commonly used buffering technique to determine which crashes are associated with a specific target road segment does not associate crashes that are closer to a target road segment with any additional weight, even though it is likely to be more greatly associated with the characteristics of the target location. Additionally, the commonly used equivalent property damage only (EPDO) crash weight method has been found to weigh fatal crashes significantly more than serious injury crashes, even if the difference between the two outcomes was a single factor. This study proposes more robust crash weighting techniques for use in high-risk location identification using an application of a novel horizontal curve dataset. Specifically, a heteroscedastic censored regression approach was used to investigate the impact of different crash proximity weighting techniques and crash severity weighting methods on model outcomes. The results demonstrate that the use of a linear distance weighting factor used in conjunction with the buffering technique as well as a less precise EPDO weighting factor method results in more robust safety analysis outcomes. The improved results have the potential to improve hot spot identification and resource allocation at both the federal and regional levels by employing models that more accurately link specific crash segments with contributing crash characteristics.