Rapid activation by photolysis of nitr-5 in skinned fibres of the striated adductor muscle from the scallop.
Photolysis of nitr-5, a caged calcium molecule, has been used for rapid activation of skinned fibre bundles of a myosin-regulated muscle, the striated adductor of the scallop, Pecten maximus. Chemically skinned fibre bundles (diameter 70-200 microns) were equilibrated in solutions containing 1-3 mM nitr-5 (pCa 6.1) and then activated by ultraviolet laser pulse (25 ns). Pulse energies of 60-95 mJ gave contractions of over 90% maximum tension and a mean half-time for tension rise of 43 ms (n = 4) at 12 degrees C. Electrically stimulated bundles of intact fibres develop a tetanus with a rise half-time of 60.2 ms at 10 degrees C (n = 5) (Rall, J.A. (1981) J. Physiol. 321, 287-295, and personal communication). At lower pulse energies the skinned fibres gave smaller amplitude contractions with slower rates of rise (up to 260 ms half-time). In addition, a slower component of tension development (mean rise half-time 13.3 s) was often observed. In ATP-free solutions containing hexokinase and glucose, rigour tension developed with a delayed onset. Rapid release of ATP (0.47-0.59 mM) from photolysis of caged ATP (2 mM) at pCa 4.5 then caused a rapid contraction with a mean half-time for tension development of 17 ms (n = 4). The fast activation rates obtained by the photorelease of Ca2+ from nitr-5 are similar to those obtained with skinned skeletal fibres of actin-regulated muscle. The results imply that the rate-limiting step in excitation-contraction coupling of the scallop muscle is not the increase in sarcoplasmic Ca2+, but rather the activation of the muscle in response to this increase. The half-times of ATP-induced contractions at pCa 4.5 suggest that in a contraction activated by a rapid Ca2+ jump the process comprising ATP hydrolysis and cross-bridge cycling occurs at a somewhat faster rate than the Ca2(+)-dependent activation process which precedes it.


The limitations of kinetic perimetry in early scotoma detection.
When tested by Goldmann kinetic perimetry ten patients with glaucomatous optic disc cupping had no visual field defects, but visual field defects were detected when tested by threshold static perimetry on the Tubingen perimeter. To improve the detection ability of kinetic perimetry, we suggest placing the interisopter spot-checks close together or expanding the central 30 degrees plotting area.


Pediatric Tracheal Lobular Capillary Hemangioma: A Case Report and Review of the Literature.
Chronic cough is a common complaint in the pediatric population and can have many different etiologies. We present a rare case of a tracheal lobular capillary hemangioma (LCH), also known as pyogenic granuloma, causing chronic cough in a child. In this case, the tracheal LCH was managed successfully with laser ablation. A review of the literature reveals only 2 other reported pediatric cases of tracheal LCH. Laryngoscope, 131:1729-1731, 2021.


Optimization and a Kinetic Study on the Acidic Hydrolysis of Dialkyl α-Hydroxybenzylphosphonates.
The two-step acidic hydrolysis of α-hydroxybenzylphosphonates and a few related derivatives was monitored in order to determine the kinetics and to map the reactivity of the differently substituted phosphonates in hydrolysis. Electron-withdrawing substituents increased the rate, while electron-releasing ones slowed down the reaction. Both hydrolysis steps were characterized by pseudo-first-order rate constants. The fission of the second P-O-C bond was found to be the rate-determining step.


Lipopolysaccharide-Induced CD300b Receptor Binding to Toll-like Receptor 4 Alters Signaling to Drive Cytokine Responses that Enhance Septic Shock.
Receptor CD300b is implicated in regulating the immune response to bacterial infection by an unknown mechanism. Here, we identified CD300b as a lipopolysaccharide (LPS)-binding receptor and determined the mechanism underlying CD300b augmentation of septic shock. In vivo depletion and adoptive transfer studies identified CD300b-expressing macrophages as the key cell type augmenting sepsis. We showed that CD300b, and its adaptor DAP12, associated with Toll-like receptor 4 (TLR4) upon LPS binding, thereby enhancing TLR4-adaptor MyD88- and TRIF-dependent signaling that resulted in an elevated pro-inflammatory cytokine storm. LPS engagement of the CD300b-TLR4 complex led to the recruitment and activation of spleen tyrosine kinase (Syk) and phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K). This resulted in an inhibition of the ERK1/2 protein kinase- and NF-κB transcription factor-mediated signaling pathways, which subsequently led to a reduced interleukin-10 (IL-10) production. Collectively, our data describe a mechanism of TLR4 signaling regulated by CD300b in myeloid cells in response to LPS.


ACT for Lupus: Pilot Feasibility and Acceptability Study of a Novel Web-Based Acceptance and Commitment Therapy Program for Patients With Lupus.
Acceptance and commitment therapy (ACT) has demonstrated effectiveness in addressing symptoms of anxiety and depression, frequently experienced by patients with systemic lupus erythematosus (SLE). The goal of this pilot study was to develop and assess the feasibility and acceptability of a novel web-based ACT skills training program tailored for patients with lupus: ACT for Lupus. The program served as a complementary approach to support the management of symptoms and stressors during the COVID-19 pandemic. This study employed a single-group pretest-posttest design. Participants with a diagnosis of SLE were primarily recruited through an institutional healthcare system between November 25, 2020 and December 17, 2020 and through an online national lupus organization listserv. Participants were invited to attend two 1-hour webinars delivered over a 2-week period. Educational ACT-based content was tailored for patients with lupus and delivered by an experienced clinical psychologist specializing in ACT. Surveys assessed patient-reported outcomes of anxiety, depression, and quality of life and evaluated program feedback, usability, and satisfaction. A total of 83 participants submitted the baseline survey, with 21 participants designated as study completers. The program was well received by participants, who reported feasibility and acceptability of the intervention, as reflected by high usability ratings. Participants reported favorable experiences with the program. Feedback included suggestions to include additional lupus-tailored content, increase the range and scope of sessions and activities, and improve program flexibility and availability to avoid scheduling conflicts. This study provides preliminary evidence for an adapted, ACT-based virtual skills training program as a feasible and acceptable intervention to support the well-being of patients with lupus.


Role of nanotechnology in HIV/AIDS vaccine development.
HIV/AIDS is one of the worst crises affecting global health and influencing economic development and social stability. Preventing and treating HIV infection is a crucial task. However, there is still no effective HIV vaccine for clinical application. Nanotechnology has the potential to solve the problems associated with traditional HIV vaccines. At present, various nano-architectures and nanomaterials can function as potential HIV vaccine carriers or adjuvants, including inorganic nanomaterials, liposomes, micelles and polymer nanomaterials. In this review, we summarize the current progress in the use of nanotechnology for the development of an HIV/AIDS vaccine and discuss its potential to greatly improve the solubility, permeability, stability and pharmacokinetics of HIV vaccines. Although nanotechnology holds great promise for applications in HIV/AIDS vaccines, there are still many inadequacies that result in a variety of risks and challenges. The potential hazards to the human body and environment associated with some nano-carriers, and their underlying mechanisms require in-depth study. Non-toxic or low-toxic nanomaterials with adjuvant activity have been identified. However, studying the confluence of factors that affect the adjuvant activity of nanomaterials may be more important for the optimization of the dosage and immunization strategy and investigations into the exact mechanism of action. Moreover, there are no uniform standards for investigations of nanomaterials as potential vaccine adjuvants. These limitations make it harder to analyze and deduce rules from the existing data. Developing vaccine nano-carriers or adjuvants with high benefit-cost ratios is important to ensure their broad usage. Despite some shortcomings, nanomaterials have great potential and application prospects in the fields of AIDS treatment and prevention.


Respiratory burst of circulating polymorphonuclear leukocytes and plasma elastase levels in patients with inflammatory bowel disease in remission.
The activation of circulating polymorphonuclear leukocytes was determined in terms of superoxide radical generation and granulocyte elastase release in untreated patients with ulcerative colitis (N = 10) and Crohn's disease (N = 9) in remission and in control subjects (N = 10). Superoxide radical generation was determined by monitoring spectrophotometrically the reduction of ferricytochrome, after stimulation of cells with phorbol myristate acetate. Plasma elastase concentration was measured by a solid-phase enzyme immunoassay technique as the complex with alpha-1-proteinase inhibitor. Superoxide formation by polymorphonuclear leukocytes from patients with ulcerative colitis and Crohn's disease was significantly lower compared with controls [median (range) nmol/min/mg protein: Crohn's disease 7.8 (7.1-9.6); ulcerative colitis 8.25 (7.4-10.3); controls 14.7 (13.6-15.8)] (P < 0.001), while no difference was found between the two groups of patients. In contrast plasma elastase levels in patients with ulcerative colitis and Crohn's disease were similar to that of controls. This defective respiratory burst of polymorphonuclear leukocytes in patients with inflammatory bowel disease in remission, in absence of an altered degranulation, could represent an important factor for the pathogenesis of these diseases.


Polygenic risk scores for prediction of breast cancer in Korean women.
Polygenic risk scores (PRSs) for breast cancer, developed using European and Asian genome-wide association studies (GWAS), have been shown to have good discrimination in Asian women. However, prospective calibration of absolute risk prediction models, based on a PRS or PRS combined with lifestyle, clinical and environmental factors, in Asian women is limited. We consider several PRSs trained using European and/or Asian GWAS. For each PRS, we evaluate the discrimination and calibration of three absolute risk models among 41 031 women from the Korean Cancer Prevention Study (KCPS)-II Biobank: (i) a model using incidence, mortality and risk factor distributions (reference inputs) among US women and European relative risks; (ii) a recalibrated model, using Korean reference but European relative risks; and (iii) a fully Korean-based model using Korean reference and relative risk estimates from KCPS. All Asian and European PRS improved discrimination over lifestyle, clinical and environmental (Qx) factors in Korean women. US-based absolute risk models overestimated the risks for women aged ≥50 years, and this overestimation was larger for models that only included PRS (expected-to-observed ratio E/O = 1.2 for women <50, E/O = 2.7 for women ≥50). Recalibrated and Korean-based risk models had better calibration in the large, although the risk in the highest decile was consistently overestimated. Absolute risk projections suggest that risk-reducing lifestyle changes would lead to larger absolute risk reductions among women at higher PRS. Absolute risk models incorporating PRS trained in European and Asian GWAS and population-appropriate average age-specific incidences may be useful for risk-stratified interventions in Korean women.


17beta-Estradiol inhibits cytokine induction of the human E-selectin promoter.
Estradiol has been shown to decrease levels of the cell adhesion molecule E-selectin in cultured cells and in women on hormone replacement therapy. We set out to determine if the mechanism of estradiol action on E-selectin is at the level of its promoter. It was found that estradiol repressed the cytokine-stimulated induction of luciferase activity driven by the human E-selectin promoter in a reporter plasmid (hE-sel-LUC) in co-transfected human hepatoma cells (Hep G2) and human umbilical cord endothelial cells (ECV-304). Repression by estradiol was dependent on the presence of transfected estrogen receptor (ER) alpha or beta expression vectors. The ER antagonist ICI-182,780 blocked the repression by estradiol, confirming the receptor-dependence of the effect. The intact DNA-binding domain of ERalpha was required for estradiol repression of the cytokine-induced stimulation of the promoter in each cell line as demonstrated by the inability of an ER construct with two point mutations in the DNA-binding domain to inhibit reporter activity. Mutation of the NFK-B site at -94 to -85 within the E-selectin promoter led to less stimulation of hE-sel-LUC by interleukin one beta (IL-1beta). Estradiol did not inhibit this IL-1beta stimulated luciferase activity, indicating that the NFK-B site is necessary for ER-mediated inhibition of this promoter. Mutation of the AP-1 site at -500 to -494 within the E-selectin promoter had no effect on the ability of IL-1beta to stimulate its transcription, and estradiol repressed this activation in an ER-dependent manner with identical efficacy and potency in comparison with the wild-type promoter. Therefore, the E-selectin promoter is down-regulated by estradiol working through either ERalpha or ERbeta and requires the NFK-B site at -94 to -85 within the promoter.


Single-cell RNA-seq transcriptomic landscape of human and mouse islets and pathological alterations of diabetes.
Single-cell RNA sequencing has paved the way for delineating the pancreatic islet cell atlas and identifying hallmarks of diabetes. However, pathological alterations of type 2 diabetes (T2D) remain unclear. We isolated pancreatic islets from control and T2D mice for single-cell RNA sequencing (scRNA-seq) and retrieved multiple datasets from the open databases. The complete islet cell landscape and robust marker genes and transcription factors of each endocrine cell type were identified. GLRA1 was restricted to beta cells, and beta cells exhibited obvious heterogeneity. The beta subcluster in the T2D mice remarkably decreased the expression of Slc2a2, G6pc2, Mafa, Nkx6-1, Pdx1, and Ucn3 and had higher unfolded protein response (UPR) scores than in the control mice. Moreover, we developed a Web-based interactive tool, creating new opportunities for the data mining of pancreatic islet scRNA-seq datasets. In conclusion, our work provides a valuable resource for a deeper understanding of the pathological mechanism underlying diabetes.


Tunable Si Dangling Bond Pathway Induced Forming-Free Hydrogenated Silicon Carbide Resistive Switching Memory Device.
With the coming of the big data age, the resistive switching memory (RSM) of three-dimensional (3D) high density shows a significant application in information storage and processing due to its simple structure and size-scalable characteristic. However, an electrical initialization process makes the peripheral circuits of 3D integration too complicated to be realized. Here a new forming-free SiCx:H-based device can be obtained by tuning the Si dangling bond conductive channel. It is discovered that the forming-free behavior can be ascribed to the Si dangling bonds in the as-deposited SiCx:H films. By tuning the number of Si dangling bonds, the forming-free SiCx:H RSM exhibits a tunable memory window. The fracture and connection of the Si dangling bond conduction pathway induces the switching from the high-resistance state (HRS) to the low-resistance state (LRS). Our discovery of forming-free SiCx:H resistive switching memory with tunable pathway opens a way to the realization of 3D high-density memory.


Evaluation of the California Mastitis Test as a precalving treatment selection tool for Holstein heifers.
The objective of this study was to evaluate the California Mastitis Test (CMT) and a portable electrical conductivity meter for diagnosing precalving intramammary infection (IMI) in Holstein heifers. A total of 428 dairy heifers from 23 dairy herds were enrolled between 6 and 12 days before the expected calving date from June 2002 to June 2003. Mammary secretions were tested by both diagnostic methods and by bacterial culture for evidence of IMI. California Mastitis Test was considered negative if the score was negative, trace or 1 and was considered positive otherwise. Two cut-off points were evaluated for milk electrical conductivity (>5 and >6.5 mS/cm). From this study, an overall proportion of 69% of heifers had precalving IMI and the overall heifer prevalence of major pathogen IMI was 16.8%. At the quarter level, sensitivity and specificity of CMT (68.9% and 68.4%, respectively) and milk conductivity >5 mS/cm (41.0% and 65.2%, respectively) or >6.5 mS/cm (25.2% and 83.3%, respectively) to identify all IMI were low. However, the heifer level sensitivity and specificity of CMT for major pathogens were 91.0% (81.5-96.6) and 27.5% (22.8-32.6), respectively. Using a cut-off point of 5 mS/cm, the heifer level sensitivity and specificity for major pathogens was 68.7% (56.2-79.4) and 44.1% (38.7-49.6), respectively. A conductivity cut-off value of 6.5 mS/cm decreased the sensitivity and increased the specificity to 53.7% (41.1-66.0) and 59.5% (54.0-64.8), respectively. California Mastitis Test and milk electrical conductivity are not good predictors of major pathogen IMI in heifers during the last 2 weeks before calving. However, the negative predictive values at quarter or heifer level were high and the heifer false negative rate was 6-14% using CMT or conductivity, respectively. Therefore, these measures could be useful for screening out heifers or quarters that are unlikely to have a major pathogen IMI.


Targeting inflammation, autophagy, and apoptosis by troxerutin attenuates methotrexate-induced renal injury in rats.
Troxerutin, a bioflavonoid with marked immune-modulatory and antioxidant features, has been proven to ameliorate experimental cardiotoxicity, hepatotoxicity, and neurodegeneration. However, its impact on methotrexate (MTX)-induced nephrotoxicity has not been investigated. In the current work, we aimed to investigate the potential of troxerutin to combat MTX-triggered renal injury, exploring immune cell infiltration, inflammation, autophagy, and apoptosis, with emphasis on the HMGB1/RAGE/NF-κB, AMPK/mTOR, and Nrf2/HO-1 pathways. Troxerutin (150 mg/kg/day) was administered by oral gavage and the renal tissues were examined with the aid of biochemical assays, ELISA, histology, and immunohistochemistry. Troxerutin mitigated MTX-induced renal dysfunction by significantly lowering creatinine, BUN, and KIM-1 alongside immune-cell infiltration and histopathologic aberrations. These favorable effects were mediated by inhibition of HMGB1/RAGE/NF-κB cascade via downregulating the protein expression of HMGB1, RAGE, and nuclear NF-κBp65 alongside its downstream signals, including COX-2 and TNF-α. Moreover, troxerutin activated the autophagy flux as evidenced by upregulating renal Beclin 1, lowering p62 SQSTM1 accumulation, and activation of AMPK/mTOR pathway, seen by increasing p-AMPK/total AMPK and lowering p-mTOR/total mTOR signals. In tandem, troxerutin combated renal apoptotic changes as proven with lowering caspase-3 activity, Bax expression, and Bax/Bcl-2 ratio and upregulating the proliferation signal PCNA. Additionally, the oxidative insult was attenuated by troxerutin, as evidenced by lowering NOX-1 and lipid peroxides, replenishing GSH, GPx, and SOD antioxidants, and activating Nrf2/HO-1 pathway. Troxerutin attenuated MTX-triggered renal injury via inhibition of inflammation and apoptosis alongside activation of autophagy. Thus, it may serve as an adjunct modality for the management of MTX-linked nephrotoxicity.


Integrated care pathway: the prevention and management of pressure ulcers.
The introduction of this pathway for the prevention and management of pressure ulcers was influenced by the development of the pathway for the management of a patient with a fractured neck of femur. Systematic documentation of preventative measures for patients with a fractured neck of femur helped reduce the incidence of pressure ulcers in this high-risk group. The original draft was informed by evidence-based clinical practice guidelines. This draft of the pathway was piloted, reviewed and changed in line with current evidence and comments from users. It was invaluable to pilot the pathway and review as necessary in order to make this a true workable document that would enable continuity of care, help prevent hospital-acquired pressure ulcers and be an effective teaching aid. A trust-wide retrospective audit of documentation for the prevention and management of pressure ulcers has been conducted, which found that the pathway facilitated a more concise approach to inform care than standard documentation. A further audit has been undertaken in the care of the elderly directorate which has demonstrated various ways in which the pathway had been completed. Challenges remain for the effective use of this documentation. Further education needs to be completed in order to promote multiprofessional use of the pathway. The pathway is now being integrated into others, which will increase awareness and involvement of other professional groups in this important aspect of patient care. Ongoing audit will take place to assess the impact upon patient outcomes.


No Evidence of Sexual Risk Compensation Following PrEP Initiation Among Heterosexual HIV Serodiscordant Couples in Kenya and Uganda.
Recent studies among men who have sex with men suggest that sexual behaviors associated with risk of sexually transmitted infections increase following initiation of pre-exposure prophylaxis (PrEP) for HIV prevention. We used longitudinal data from HIV-uninfected participants (n = 1013) enrolled in an open-label study of PrEP delivered to Ugandan and Kenyan heterosexual HIV serodiscordant couples to understand the association between PrEP initiation and HIV risk-related sexual behaviors among these couples. In the month following PrEP initiation, the mean number of monthly sex acts within couples decreased from 7.9 to 6.9 (mean difference: - 1.1; 95% CI - 1.5, - 0.7) and the proportion of couples having condomless sex decreased from 65% to 32% (percentage point change: - 33%; 95% CI - 37%, - 30%); these behaviors then remained relatively constant over 2 years. We found no evidence of sexual risk compensation following PrEP initiation within African serodiscordant couples. However, roughly a third of couples continued to engage in condomless sex during follow up, emphasizing the importance of continued PrEP use to sustain HIV protection.


Cochlear Implantation in Children with Otitis Media.
One of the concerns during the cochlear implant candidacy process is the presence of chronic otitis media which could delay the implantation process. The aim of this study was to evaluate the surgical difficulties and the long-term complications in children with otitis media and to examine whether it is necessary to delay the implantation until the infection is resolved. The study used a comparative retrospective design based on chart review of all patients who received their implant(s) from January to December of 2012. A total of 200 patients were identified and were followed for 4 years post surgery. Patients were divided into three groups based on their history of otitis media (non-otitis media, chronic otitis media with effusion, and acute otitis media). Data included long-term complications, operative time and duration from first clinical visit to the time until implantation was received. None of study participants had long-term complications during the study period. The operative time was longer in the acute otitis media group with a difference of 45 min. The average delay in cochlear implantation due to the presence of otitis media in chronic group was more than 5 months. Pediatric patients with otitis media could be implanted in one stage safely and effectively.


4-Allyl-5-pyridin-4-yl-2,4-dihydro-3H-1,2,4-triazole-3-thione: synthesis, experimental and theoretical characterization.
The title molecule, 4-allyl-5-pyridin-4-yl-2,4-dihydro-3H-1,2,4-triazole-3-thione (C(10)H(10)N(4)S), was synthesized and characterized by IR-NMR spectroscopy and single-crystal X-ray diffraction. The compound crystallizes in the monoclinic space group is P2(1)/c, a=8.006(5)Å, b=15.363(5)Å, c=8.936(5)Å, β=104.441(5)° and V=1064.4(10)Å(3), F(000)=456, D(x)=1.362 g/cm(3). In addition to the molecular geometry from X-ray experiment, the molecular geometry, vibrational frequencies, gauge including atomic orbital (GIAO) (1)H and (13)C chemical shift values of the title compound in the ground state have been calculated using the Hartree-Fock (HF) and density functional method (DFT/B3LYP) with 6-31G(d) basis set. To determine conformational flexibility, molecular energy profile of the title compound was obtained by HF/6-31G(d) and (DFT/B3LYP) calculations with respect to selected degree of torsional freedom, which was varied from -180° to +180° in steps of 10°. Besides, molecular electrostatic potential (MEP), frontier molecular orbitals (FMO), and several thermodynamic properties were performed by the HF and DFT methods.


Sexual dimorphism and fruit production in a dioecious understory tree, Ilex opaca Ait.
This study suggested that sexual selection is potentially an important factor in the maintenance of dioecy in the American holly, Ilex opaca (Aquifoliaceae). Sexual dimorphisms in flower production and phenology were highly significant in this understory tree. On average, individual males produced 7.4 times as many flowers as did female trees. Staminate flowers lasted only a single day, whereas pistillate flowers lasted 3-4 days, during which they showed no significant decline in their ability to produce fruit after pollination. Individual male trees opened their flower buds asynchronously during the season, maximizing the number of days they were in flower. Individual females opened their buds more synchronously, maximizing their floral display at one point in time. Females produced fruits in numbers that were somewhat less than proportional to their flower production. Fruit development was initiated from only 38.9% and 69.5% of pistillate flowers in 1987 and 1988, respectively. By the time of ripening, an average female had lost 62.3%, 24.3%, and 11.1% of its initial fruit crop in 1986, 1987, and 1988, respectively. The proportion of fruit lost in 1986 was independent of the number of fruit that initially began development. In 1988, artifically supplementing pollen to a large number of flowers failed to increase either fruit or seed production relative to control branches with unsupplemented flowers. This suggested that resource levels were likely more important than pollen availability in limiting female reproductive success. These observations on I. opaca were consistent with the expectations for a population in which male reproductive success continues to benefit from continued pollinator service and female reproductive success does not.


Reflex epilepsy.
We report 3 cases of reflex epilepsy, 2 of them triggered by reading (reading epilepsy) and one by mathematical calculation (epilepsy arithmetics). Electroencephalographic abnormalities predominated in the dominant hemisphere. The pathophysiologic mechanisms involved in this rare form of epilepsy are commented on, particularly the role played by the association of mental concentration and emotional strain. A possible deficiency in GABA-ergic inhibitory synapses would cause a deficit in cortical inhibition. Different drugs have been used successfully in patients with reflex epilepsy. Psychotherapy relieves anxiety and has been shown to be an effective complement to medical treatment.


Tracheal intubation in children with Morquio syndrome using the angulated video-intubation laryngoscope.
There are a number of syndromes with proven or suspected instability of the cervical spine especially in pediatric patients. It is a challenge for the anesthesiologist to intubate these patients with as little movement of the cervical spine as possible. A new device to facilitate this task is the angulated video-intubation laryngoscope (AVIL). The AVIL is a curved endoscopic intubation laryngoscope with angulated distal tip. The video-view from the distal blade tip improves glottic visualization during difficult direct laryngoscopy. We report three sisters with Morquio syndrome scheduled for otorhinolaryngology surgery the same day. Two of them had radiologically suspected cervical spine instability. Tracheal intubation was planned with careful direct laryngoscopy under manual in-line stabilization of the neck and head by an assistant. Direct visualization of the larynx using a Miller blade No. 2 was impaired in two of the three children in whom the cervical spine was immobilized. They were both successfully intubated under endoscopic control using the AVIL. The AVIL may become a helpful device to aid endotracheal intubation in patients when cervical spine immobilization impairs direct laryngoscopy.


Acidic infusion in early reperfusion affects the activity of antioxidant enzymes in postischemic isolated rat heart.
Acidic perfusion (AP) performed at the onset of reperfusion (i.e., acid postconditioning) is cardioprotective. We investigated the effect of AP on postischemic cardiac function and on the activity of endogenous superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase. The role of exogenous CAT or SOD on AP cardioprotection was also investigated. Phosphorylation of redox-sensitive survival kinases (protein kinase C [PKC] ε and extracellular signal-regulated kinase [ERK] 1/2) was also checked. Isolated rat hearts underwent ischemia and reperfusion (I/R) for 30 and 120 min, respectively. AP was obtained by lowering [HCO3(-)] in the perfusion buffer. Infarct size and left ventricular pressure were measured. Protocols include I/R only, I/R plus acidic perfusion in early reperfusion (I/R + AP), and I/R plus AP and CAT (I/R + AP + CAT) or SOD (I/R + AP + SOD). I/R + SOD and I/R + CAT additional hearts served as controls. AP and/or antioxidants were given in the initial 3 min of reperfusion. Enzyme activities were studied in postischemic phase (seventh minute of reperfusion) in I/R or I/R + AP and Sham (buffer-perfused) hearts. AP with (I/R + AP + CAT or I/R + AP + SOD) or without (I/R + AP) antioxidant enzymes resulted in a larger reduction of infarct size compared with I/R, I/R + SOD, or I/R + CAT. Compared with I/R, the postischemic systolic and diastolic recoveries of the cardiac function were markedly improved by the addition of AP and a lesser extent by AP + SOD or AP + CAT. AP increased the postischemic activity of CAT and lowered that of SOD and glutathione peroxidase compared with I/R only. Also, the phosphorylation and activity of ERK1/2 and PKCε were increased by AP. Acid postconditioning affects the activity of endogenous antioxidant enzymes, activates ERK1/2-PKCε pathways, and protects against myocardial I/R injury. The combination of AP and exogenous SOD or CAT still provides cardioprotection. It is likely that intracellular (not extracellular) redox condition plays a pivotal role in acidic protection.


Patient And Nurse Experience Of Using Somatostatin Analogues To Treat Gastroenteropancreatic Neuroendocrine Tumors: Results Of The Somatostatin Treatment Experience Trial (STREET).
Evaluate patients' and nurses' experiences, including injection problem frequency, with the somatostatin analogues (SSAs) lanreotide autogel® (Somatuline® autogel®, deep subcutaneous) and octreotide long-acting release (LAR) (Sandostatin® LAR®, intramuscular) when treating gastroenteropancreatic neuroendocrine tumors (GEP-NETs). An observational, cross-sectional study across 2 NET centers in Sweden. Questionnaires based on participants' most recent injection experience were sent to patients with GEP-NETs treated with octreotide or lanreotide, and to nurses administering these treatments. Nurses were identified via patients completing their questionnaires. Resource use was sourced from Swedish prescription registry records. The planned sample size was 200, based on an estimated proportion of 0.50 and ±7% precision. 119/156 patients (n=53, lanreotide; n=66, octreotide) and 43/53 nurses (n=22, lanreotide; n=21, octreotide) completed questionnaires. Despite smaller recruitment than planned, the endpoint precision was ±9% with 119 participants, and still considered reasonable. More octreotide-treated patients reported problems (18% vs none; P=0.001) and experienced moderate-to-high anxiety pre-injection (11% vs 2%). Patients had similar physical HRQoL scores overall (Short Form-12 mean composite scores: physical: 39.4 vs 37.6; mental: 50.7 vs 49.6). The mean number of lanreotide and octreotide doses dispensed per year were 11.1 and 12.6, respectively (P<0.05). In the lanreotide group, 28% self-injected, while 29% were not aware they could self-inject. In the octreotide group, 3% self-injected and 73% were unaware of the availability of an SSA for self-injection. Most patients (61%) felt well-informed about their disease and treatment. Nurses were generally experienced and felt confident and well-informed about giving SSA injections; however, only 12% felt well-informed about the disease and treatment. Those treated with lanreotide reported fewer injection problems and experienced less pre-injection anxiety than those treated with octreotide. SSA choice did not appear to affect patients' HRQoL. Some patients treated with octreotide were unaware of an SSA with the flexibility of self-injection.


SERS assessment of the cancer-specific methylation pattern of genomic DNA: towards the detection of acute myeloid leukemia in patients undergoing hematopoietic stem cell transplantation.
In this label-free surface-enhanced Raman scattering (SERS) study of genomic DNA, we demonstrate that the cancer-specific DNA methylation pattern translates into specific spectral differences. Thus, DNA extracted from an acute myeloid leukemia (AML) cell line presented a decreased intensity of the 1005 cm-1 band of 5-methylcytosine compared to normal DNA, in line with the well-described hypomethylation of cancer DNA. The unique methylation pattern of cancer DNA also influences the DNA adsorption geometry, resulting in higher adenine SERS intensities for cancer DNA. The possibility of detecting cancer DNA based on its SERS spectrum was validated on peripheral blood genomic DNA samples from n = 17 AML patients and n = 17 control samples, yielding an overall classification of 82% based on the 1005 cm-1 band of 5-methylcytosine. By demonstrating the potential of SERS in assessing the methylation status in the case of real-life DNA samples, the study paves the way for novel methods of diagnosing cancer. Graphical abstract.


Internalization of the urokinase-plasminogen activator inhibitor type-1 complex is mediated by the urokinase receptor.
The role of the urokinase receptor (uPAR) in the internalization of the urokinase-plasminogen activator inhibitor type-1 (uPA.PAI-1) complex has been investigated. First, exploiting the species specificity of uPA binding, we show that mouse LB6 cells (that express a mouse uPAR) were unable to bind or degrade the human uPA.PAI-1 complex. On the other hand, LB6 clone 19 cells, which express a transfected human uPAR, degraded uPA.PAI-1 complexes with kinetics identical to the human monocytic U937 cells. We also show by immunofluorescence experiments with anti-uPA antibodies that in LB6 clone 19 cells, the uPA.PAI-1 complex is indeed internalized. While at 4 degrees C uPA fluorescence was visible at the cell surface, shift of the temperature to 37 degrees C caused a displacement of the immunoreactivity to the cytoplasmic compartment, with a pattern indicating lysosomal localization. If uPA.PAI-1 internalization/degradation is mediated by uPAR, inhibition of uPA.PAI-1 binding to uPAR should block degradation. Three different treatments, competition with the agonist amino-terminal fragment of uPA, treatment with a monoclonal antibody directed toward the binding domain of uPAR or release of uPAR from the cell surface with phosphatidylinositol-specific phospholipase C completely prevented uPA.PAI-1 degradation. The possibility that a serpin-enzyme complex receptor might be primarily or secondarily involved in the internalization process was excluded since a serpin-enzyme complex peptide failed to inhibit uPA.PAI-1 binding and degradation. Similarly, complexes of PAI-1 with low molecular mass uPA (33 kDa uPA), which lacks the uPAR binding domain, were neither bound nor degraded. Finally we also show that treatment of cells with uPA.PAI-1 complex caused a specific but partial down-regulation of uPAR. A similar result was obtained when PAI-1 was allowed to complex to uPA that had been previously bound to the receptor. The possibility therefore exists that the entire complex uPA.PAI-1-uPAR is internalized. All these data allow us to conclude that internalization of the uPA.PAI-1 complex is mediated by uPAR.


Introducing titratable water to all-atom molecular dynamics at constant pH.
Recent development of titratable coions has paved the way for realizing all-atom molecular dynamics at constant pH. To further improve physical realism, here we describe a technique in which proton titration of the solute is directly coupled to the interconversion between water and hydroxide or hydronium. We test the new method in replica-exchange continuous constant pH molecular dynamics simulations of three proteins, HP36, BBL, and HEWL. The calculated pKa values based on 10-ns sampling per replica have the average absolute and root-mean-square errors of 0.7 and 0.9 pH units, respectively. Introducing titratable water in molecular dynamics offers a means to model proton exchange between solute and solvent, thus opening a door to gaining new insights into the intricate details of biological phenomena involving proton translocation.


Induction of secretory antibodies in humans following ingestion of Streptococcus mutans.
1) Ingestion of Streptococcus mutans antigen by human volunteers induced the selective appearance of antibodies in saliva and tears but not in serum. 2) These antibodies were associated with the IgA class as determined by enhanced or blocked agglutination, immunofluorescence and gel filtration. 3) A second series of antigen ingestion resulted in a secondary response in these secretions characterized by an earlier appearance of antibodies which reached higher titers than in the primary response.


Observation of Changes in Helicobacter pylori Antigen and Antibody Positivity According to Non-Invasive Tests Before and After Helicobacter pylori Eradication Therapy in Symptomatic Patients.
Non-invasive tests can help with the diagnosis of Helicobacter pylori (H. pylori) infection and in determining patient prognosis following H. pylori eradication therapy. The aim of the study was to detect H. pylori antigens in the stool in symptomatic patients and to observe changes in the antigen test results following H. pylori eradication therapy. A prospective study was conducted. Blood, urine and stool samples were collected from 62 dyspeptic patients. Anti-H. pylori IgM and IgG antibodies were detected in the serum by ELISA, anti-H. pylori IgG antibodies were detected in the urine by ICT and H. pylori antigens were detected in the stool by ELISA. Among the 62 patients, 39 (62.90%) were positive with all three methods. These 39 patients were asked to complete a 2-week course of medication and return after 6 weeks following completion of therapy to undergo repeated tests. In total, 3 dropped out of the study. Among the 62 dyspeptic patients, 41 (66.13%) were positive for serum IgG according to ELISA, 39 (62.90%) were positive for urine IgG according to ICT, 8 (12.90%) were positive for serum IgM according to ELISA, and 42 (67.74%) were positive for HpSA according to ELISA. After eradication therapy, 18 (50.00%) patients were positive for serum IgG, 19 (52.78%) were positive for urine IgG, 4 (11.11%) were positive for serum IgM and 5 (13.88%) were positive for HpSA. The difference in HpSA positivity before and after eradication therapy was statistically significant (P <0.05). This study involved non-invasive procedures that can be used as first-line screening tools for the detection of active H. pylori infection to observe the role of HpSA test in diagnosis and assessment of prognosis following eradication therapy for H. pylori.


Selection of a macaque Fab with framework regions like those in humans, high affinity, and ability to neutralize the protective antigen (PA) of Bacillus anthracis by binding to the segment of PA between residues 686 and 694.
Human anthrax infection cannot always be treated successfully by antibiotics, as highlighted by recent bioterrorist attacks; thus, adjunct therapies are clearly needed for the future. There is a particular need to further develop adjunct therapies that can neutralize secreted toxins, such as antibodies directed towards the 83-kDa protective antigen (PA(83)). In the absence of human donors, we immunized a macaque (Macaca fascicularis) with PA(83) to obtain such antibodies suitable as an adjunct therapy for human anthrax infection. By using bone marrow as a template, we PCR amplified specific Fab-encoding genes and cloned them as an immune library (10(7) clones). We isolated a high-affinity (equilibrium dissociation constant [K(D)], 3.4 nM), highly neutralizing (50% inhibitory concentration, 5.6 +/- 0.13 nM) Fab (designated 35PA(83)) from this library by panning. Its epitope was localized by Pepscan analysis between residues 686 and 694 of PA(83) and is part of the region which directly interacts with the cell receptor. 35PA(83) may thus neutralize the anthrax toxin by competing directly for its receptor. The genes encoding 35PA(83) were similar to those of a human immunoglobulin germ line and were assigned to subgroups of human V, (D), or J genes by IMGT/V-QUEST analysis. The 35PA(83) framework regions were 92% identical to a representative allele of each subgroup. When compared to framework regions coded by related human germ line genes, only 2 of 74 (VH) or 75 (VK) analyzed amino acids of 35PA(83) have different chemical characteristics. A very high degree of identity with human framework regions makes 35PA(83) well suited for expression as a whole primatized immunoglobulin G and demonstrates the practicality of using macaque Fabs when immunized human plasma cell donors are not available.


[Methodologic approach to the therapeutic problem of infant and childhood obesity. Review of the literature and personal experience].
Notwithstanding the high prevalence and morbidity, current knowledge of obesity is rather rudimentary. Nevertheless recent findings have enabled us to surmount anachronistic nosographic generalizations whereby excess weight was attributed only to inappropriate and excessive food intake and the lack of physical exercise. Various studies indicate that there are complex interactions in determining weight excess: genetic, psychosocial, cultural but also biochemical factors are incriminated and it is now recognised that the aetiology of obesity is multifactorial and that, consequently, there are different types of obesity. However, in most studies concerning the therapeutic approach, it is noted that the same type of treatment is applied to groups of subjects that are probably highly heterogeneous for type of obesity. No easy or final solution to the problem is proposed as for the moment no ideal treatment exists. What is suggested is the need to attempt to identify possible factors linked to weight excess before commencing the therapeutic approach. For each single case, this could permit the use of the most appropriate treatment and make therapeutic results less frustrating.


Satisfaction with telephonic interpreters in pediatric care.
To compare satisfaction among Spanish-speaking mothers who did and did not use telephonic interpreters during pediatric visits, and to examine resident physician attitudes about telephonic interpreter use. Anonymous surveys were administered to 98 mothers limited in English proficiency and presenting for well-baby visits in an urban university hospital-affiliated practice. Pediatric visits were performed by 24 non-Spanish-proficient pediatric residents. The first 46 women (baseline cohort) received routine services, including ad-hoc interpretation or no interpretation; the second 52 women (intervention cohort) used a dual-headset telephonic interpreter service. Mothers completed postvisit interviews assessing overall satisfaction, comfort and ease of communication. Pediatric residents completed self-administered questionnaires assessing attitudes about and experience with telephonic interpretation. The intervention cohort overwhelmingly rated telephonic interpretation as "very helpful" (94%), indicating the visit would have been "harder" without the service (98%). Significantly more intervention cohort mothers reported it was "very easy" to communicate with the physician (83% vs. 22%, P < 0.01), they understood "all" that the physician told them (97% vs. 80%, P < 0.05) and they were "very satisfied" with the clinic overall (85% vs. 57%, P < 0.05). Almost all intervention cohort mothers (96%) reported a preference to use telephonic interpretation at their subsequent visit; however, only one-third of residents believed their patients would prefer to use the telephonic interpreter in the future. Mothers who used telephonic interpretation reported significantly greater communication and overall satisfaction compared to mothers in routine care. Pediatric residents substantially underestimated their patients' desire to use telephonic interpreters.


Detection of divinyl ether synthase in Lily-of-the-Valley (Convallaria majalis) roots.
Incubations of linoleic acid with cell-free preparations from Lily-of-the-Valley (Convallaria majalis L., Ruscaceae) roots revealed the presence of 13-lipoxygenase and divinyl ether synthase (DES) activities. Exogenous linoleic acid was metabolized predominantly into (9Z,11E,1'E)-12-(1'-hexenyloxy)-9,11-dodecadienoic (etheroleic) acid. Its identification was confirmed by the data of ultraviolet spectroscopy, mass spectra, (1)H NMR, COSY, catalytic hydrogenation. The isomeric divinyl ether (8E,1'E,3'Z)-12-(1',3'-nonadienyloxy)-8-nonenoic (colneleic) acid was detected as a minor product. Incubations with linoleic acid hydroperoxides revealed that 13-hydroperoxide was a preferential substrate, while the 9-hydroperoxide was utilized with lesser efficiency.


HoxA9 induces insulin-like growth factor-1 receptor expression in B-lineage acute lymphoblastic leukemia.
The homeobox (Hox) gene family encodes a group of transcription factors preferentially expressed during embryonic development and hematopoiesis. Deregulation of Hox gene expression is frequently associated with acute leukemia. HoxA9 is the most commonly overexpressed Hox gene in acute leukemia. However, little is known regarding specific pathways regulated by HoxA9 that promote the growth and survival of leukemic cells. We have generated a conditional model of HoxA9 activity in the stromal cell dependent, HoxA9 negative, pre-B-cell line B-lineage-2 (BLIN-2). Conditional HoxA9 activation in BLIN-2 resulted in increased proliferation in the presence and absence of stromal cell support. Stimulation of HoxA9 activity resulted in increased expression of the c-Myb transcription factor and induction of insulin-like growth factor-1 receptor (IGF-1R) surface expression. HoxA9-mediated proliferative effects in BLIN-2 cells were abrogated when the cells were treated with specific IGF-1R tyrosine kinase inhibitors or with an IGF-1R mAb (A12). IGF-1R expression correlated with endogenous HoxA9 expression in a small panel of mixed lineage leukemia (MLL)/AF4 cell lines. siRNA knockdown of endogenous HoxA9 expression in the MLL/AF4-positive cell line RS4;11 resulted in loss of IGF-1R expression. These data indicate that HoxA9 overexpression induces IGF-1R expression and subsequently promotes leukemic cell growth.


Serotonin 5-HT2A receptor involvement and Fos expression at the spinal level in vincristine-induced neuropathy in the rat.
We recently showed that peripheral and spinal 5-HT2A receptors (5-HT2AR) are involved in a rodent model of neuropathy induced by a nucleoside analogue reverse transcriptase inhibitor. In this paper, we show that 5-HT2AR are also involved in neuropathy induced by an anti-neoplasic drug, vincristine. Vincristine-treated rats (0.1mg/kg, daily i.p. administration for two 5-day cycles) developed thermal allodynia and mechanical hypersensitivity, which decreased in a dose-related manner after epidural injection a 5-HT2A receptor antagonist. Moreover, 5-HT2A-/- mice did not develop vincristine-induced neuropathy contrarily to their 5-HT2A+/+ littermates. In vincristine-treated rats, the number of nociceptive dorsal root ganglion cells expressing the 5-HT2AR was increased by 38%, and 5-HT2AR immunolabelling was enhanced in layers I-IV of the dorsal horn. At the EM level, a 76.3% increase in the density of 5-HT2AR immunopositive axon terminals within superficial layers of the dorsal horn was noted after vincristine treatment. Immunocytochemical study of Fos expression in vincristine-treated rats revealed a significant increase in the number of Fos-positive neurons not only in regions where nociceptive fibres terminate superficial (I-II) and deep layers (V-VI) of the spinal cord, but also in intermediate layers, suggesting that Abeta fibres could be involved in the spinal sensitization observed in this model. Double labelling experiments showed that Fos-positive neurons were endowed with 5-HT2AR immunolabelling in the dorsal horn of vincristine-treated rats. These data provide support to the idea that, in vincristine-induced neuropathy, 5-HT2AR are involved in the sensitization of peripheral nociceptors and spinal nociceptive processing.


Assessment of HER-2/neu status in breast cancer using fluorescence in situ hybridization & immunohistochemistry: Experience of a tertiary cancer referral centre in India.
Determination of HER2 status in breast cancer has become important to identify potential candidates for anti-HER2 therapy. In this study we compared fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) for the determination of HER2 status in breast cancer patients referred to a tertiary care referral centre. A total of 200 cases of invasive breast cancer were evaluated for HER2 status using IHC and FISH and results were compared. The IHC 3+ (93.9%) and IHC negative (85.9%) cases showed good concordance with the corresponding FISH results; while 66.6 per cent of IHC 2+ cases showed gene amplification by FISH. In addition, hormone receptor expression and HER2 gene status showed a statistically significant inverse association (P<0.05). These findings reaffirm IHC as a prudent first-step to screen tissue samples for HER2 status and to determine suitability for technically demanding FISH test and the dual coloured FISH as a gold standard for determination of HER2/neu status in IHC equivocal cases of breast carcinoma.


Pharmacokinetics of a valpromide isomer, valnoctamide, in healthy subjects.
The pharmacokinetics of a single 400 mg oral dose of valnoctamide (VCD) has been investigated in seven healthy, adult, male volunteers. VCD was not biotransformed rapidly to its corresponding acid valnoctic acid (VCA), unlike its isomer valpromide (VPD). It had a mean residence time of 13.2 h and a terminal half-life of 9.3 h. Throughout the study, only low plasma levels of VCA could be detected. Thus, unlike VPD, which is a pro-drug of the corresponding acid, (valproic acid, VPA). VCD appears to act as a drug in its own right, and it does not undergo similar hydrolysis. The pharmacokinetic difference may account for the different pharmacological activities of the two isomers.


Effect of carboxamide derivative IS-741 on rat spontaneous chronic pancreatitis.
This study aimed to analyze the therapeutic effects of the carboxamide derivative IS-741 on spontaneous chronic pancreatitis (CP) in the WBN/Kob rat. Four-week-old male WBN/Kob rats were fed with MB-3, and IS-741 (0.012% in MB-3) was orally administered for 12 weeks. Cytokine-induced neutrophil chemoattractant (CINC), interleukin (IL) -6, pancreatitis-associated protein (PAP), and p8 mRNAs in the pancreas were detected with semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Histological examination of the pancreas showed that hemorrhage, interstitial edema, and inflammatory cell infiltration were markedly less severe at 12 weeks, while fibrosis and acinar degeneration were also milder at 16 weeks, in the IS-741-treated rats than in untreated rats. The above-mentioned genes were significantly suppressed in IS-741-treated rats at 12 weeks. Our results show that IS-741 exerts a potentially therapeutic effect on experimental CP by suppressing the expressions of the proinflammatory cytokines (IL-6 and CINC) and pancreatic acute phase proteins (PAP and p8).


Dominant inheritance of cerebral gigantism.
Cerebral gigantism is a syndrome consisting of characteristic dysmorphic features, accelerated growth in early childhood, and variable degrees of mental retardation. Its etiology and pathogenesis have not been defined. Three families are presented with multiple affected members. The vertical transmission of the trait and equal expression in both sexes in these families indicates a genetic etiology with a dominant pattern of inheritance, probably autosomal. As in previously reported cases, extensive endocrine evaluation failed to define the pathogenesis of the accelerated growth present in this disorder.


Graph matching and deep neural networks based whole heart and great vessel segmentation in congenital heart disease.
Congenital heart disease (CHD) is one of the leading causes of mortality among birth defects, and due to significant variations in the whole heart and great vessel, automatic CHD segmentation using CT images has been always under-researched. Even though some segmentation algorithms have been developed in the literature, none perform very well under the complex structure of CHD. To deal with the challenges, we take advantage of deep learning in processing regular structures and graph algorithms in dealing with large variations and propose a framework combining both the whole heart and great vessel segmentation in complex CHD. We benefit from deep learning in segmenting the four chambers and myocardium based on the blood pool, and then we extract the connection information and apply graph matching to determine the categories of all the vessels. Experimental results on 68 3D CT images covering 14 types of CHD illustrate our framework can increase the Dice score by 12% on average compared with the state-of-the-art whole heart and great vessel segmentation method in normal anatomy. We further introduce two cardiovascular imaging specialists to evaluate our results in the standard of the Van Praagh classification system, and achieves well performance in clinical evaluation. All these results may pave the way for the clinical use of our method in the incoming future.


RNA interference targeting the platelet-derived growth factor receptor beta subunit ameliorates experimental hepatic fibrosis in rats.
Platelet-derived growth factor (PDGF) is the strongest stimulator of the proliferation of hepatic stellate cells (HSCs). PDGF receptor beta subunit (PDGFR-beta) is acquired on HSCs proliferation induced by PDGF. In this study, we aim to investigate the effect of PDGFR-beta small interference RNA (siRNA) on experimental hepatic fibrosis. We constructed a PDGFR-beta siRNA expression plasmid and investigated its effect on the activation of HSCs. Bromodeoxyuridine incorporation was performed to investigate the effect of PDGFR-beta siRNA on HSCs proliferation. A hydrodynamics-based transfection method was used to deliver PDGFR-beta siRNA to rats with hepatic fibrosis. The distribution of transgenes in the liver was observed by immunofluorescence. The antifibrogenic effect of PDGFR-beta siRNA was investigated pathologically. Platelet-derived growth factor receptor-beta subunit siRNA could significantly downregulate PDGFR-beta expression, suppress HSCs activation, block the mitogen-activated protein kinase signalling pathway and inhibit HSCs proliferation in vitro. PDGFR-beta siRNA expression plasmid could be delivered into activated HSCs by the hydrodynamics-based transfection method, and remarkably improve the liver function of the rat model induced by dimethylnitrosamine and bile duct ligation. Furthermore, the progression of fibrosis in the liver was significantly suppressed by PDGFR-beta siRNA in both animal models. Platelet-derived growth factor receptor-beta subunit siRNA may be presented as an effective antifibrogenic gene therapeutic method for hepatic fibrosis.


Status Quo of Diagnostic Procedures and Treatment of Inpatients with Infective Endocarditis at the Department of General Medicine at a University Hospital in a Suburban City in Japan: A Single-Hospital-Based Retrospective Study.
Our hospital's department of general medicine is often involved in the diagnosis and treatment of diseases that are considered by other hospitals or other departments in our hospital to be difficult to diagnose correctly. The aim of this study was to clarify how patients with infective endocarditis (IE) being admitted to our hospital's department of general medicine were examined and treated and to elucidate their prognosis compared with patients admitted to other departments. Inpatients of Saga University Hospital with definite IE from September 2007 to August 2017 were divided into 2 groups: those admitted to the general medicine department (the GM group) and those admitted to other departments (the non-GM group). Seventy-four patients were included; 17 (23%) were admitted to the general medicine department. In the GM group, the percentage of patients diagnosed with definite or suspected IE was lower (0% vs 32%, p=0.008), as was the rate of patients with echocardiographic findings that fulfilled the major modified Duke's criteria (71% vs 98%, p≤0.001), preadmission. The GM group had higher percentages of patients with back or joint pain (41% vs 9%, p=0.001) and complications, including pyogenic spondylitis (35% vs 2%, p≤0.001), deep-seated abscesses (24% vs 5%, p=0.024), pyogenic arthritis (18% vs 0%, p=0.001), and glomerulonephritis (77% vs 37%, p=0.004) than did the non-GM group. Mortality within 30 days of admission to our hospital (12% vs 14%, p=0.753) and overall in-hospital mortality (12% vs 18%, p=0.570) did not significantly differ. The general medicine department could have accurately diagnosed IE, given appropriate treatments, and obtained similar prognoses to those of IE patients treated by other departments, including cardiology and cardiovascular surgery, even in patients for whom diagnosing IE was more difficult or who had less typical echocardiographic findings preadmission.


Free antibodies-to-infliximab are biomarker for predicting the effect of dose intensification in pediatric Crohn's disease patients with secondary loss of response.
Immunogenicity to antitumor necrosis factor alpha agents, such as infliximab (IFX), may lead to therapeutic failure. This study evaluated the relationship between free and total antibodies-to-infliximab (ATIs), trough levels (TLs) of IFX, and the response to dose intensification. We performed a prospective, observational study including pediatric patients with Crohn's disease (CD) receiving IFX maintenance therapy without dose intensification. We compared clinical and laboratory outcomes according to the presence of free and total ATIs. Factors associated with response to IFX dose intensification were investigated by analyzing IFX TLs and free and total ATIs. Of the 98 patients, 9 patients had detectable free ATIs and 38 patients had total ATIs. Patients with free ATIs had significantly lower TLs (0.7 versus 5.1 µg/mL, p < 0.001) than patients without free ATIs. However, there was no difference in the IFX TLs according to the presence of total ATIs (p = 0.2523). Analysis of the 38 samples with total ATIs showed that response to dose intensification was significantly lower in patients with free ATIs than those without free ATIs (22.2% versus 65.5%, p < 0.001). In addition, free ATIs were the only factor with poor response to dose intensification [odds ratio (OR): 14.15, 95% confidence interval (CI): 1.31-151.97, p = 0.0140]. According to the receiver operating characteristic analysis, the optimal cutoff level indicating non-response to IFX dose intensification was 30.0 AU/mL for free ATIs concentration (area under curve, 0.792; 95% CI: 0.590-0.942; sensitivity, 60.0%; specificity, 96.7%; p = 0.0241). Free ATIs, but not total ATIs, have a negative impact on the course of CD. Free ATIs are potential reliable biomarker for predicting the effect of dose intensification in patients with loss of response to IFX. Future studies based on serial and proactive therapeutic drug monitoring are required in the future.


SOD3 Ameliorates Aβ25-35-Induced Oxidative Damage in SH-SY5Y Cells by Inhibiting the Mitochondrial Pathway.
This study was designed to investigate the protective effects of extracellular superoxide dismutase (SOD3) against amyloid beta (Aβ25-35)-induced damage in human neuroblastoma SH-SY5Y cells and to elucidate the mechanisms responsible for this beneficial effect. SH-SY5Y cells overexpressing SOD3 were generated by adenoviral vector-mediated infection and Aβ25-35 was then added to the cell culture system to establish an in vitro model of oxidative stress. Cell viability, the generation of intracellular reactive oxygen species (ROS), the expression and activity of antioxidant enzymes, the levels of lipid peroxidation malondialdehyde (MDA), the expression of mitochondrial apoptosis-related genes and calcium images were examined. Following Aβ25-35 exposure, SOD3 overexpression promoted the survival of SH-SY5Y cells, decreased the production of ROS, decreased MDA and calcium levels, and decreased cytochrome c, caspase-3, caspase-9 and Bax gene expression. Furthermore, SOD3 overexpression increased the expression and activity of antioxidant enzyme genes and Bcl-2 expression. Together, our data demonstrate that SOD3 ameliorates Aβ25-35-induced oxidative damage in neuroblastoma SH-SY5Y cells by inhibiting the mitochondrial pathway. These data provide new insights into the functional actions of SOD3 on oxidative stress-induced cell damage.


Anti-HIV activity of N-1-adamantyl-4-aminophthalimide.
The discovery of new leads acting via novel modes of action in the treatment of the human immunodeficiency virus (HIV), the causative agent of AIDS, remains a challenge. Along this line we synthesized and evaluated a series of N-substituted 4-aminophthalimides which were designed according to the models of thalidomide, phenytoin (PHT) and ameltolide. From a series of 24 compounds only N-1-adamantyl-4-aminophthalimide was endowed with anti-HIV-1 and -HIV-2 activity in CEM cell cultures.


MicroRNA negatively regulates NF-κB-mediated immune responses by targeting NOD1 in the teleost fish Miichthys miiuy.
Inflammation is a self-protection mechanism that can be triggered when innate immune cells detect infection. Eradication of pathogen infection requires appropriate immune and inflammatory responses, but excessive inflammatory responses can cause uncontrolled inflammation, autoimmune diseases, or pathogen dissemination. Mounting evidence has shown that microRNAs (miRNAs) in mammals act as important and versatile regulators of innate immunity and inflammation. However, miRNA-mediated regulation networks are largely unknown in inflammatory responses in lower vertebrates. Here miR-144 and miR-217 are identified as negative regulators in teleost inflammatory responses. We find that Vibrio harveyi and lipopolysaccharide (LPS) treatment significantly upregulate the expression of fish miR-144 and miR-217. Upregulated miR-144 and miR-217 suppress LPS-induced inflammatory cytokine expression by targeting nucleotide-binding oligomerization domain-containing protein 1 (NOD1), thereby avoiding excessive inflammatory responses. In addition, miR-144 and miR-217 regulate inflammatory responses through NOD1-induced nuclear factor kappa (NF-kB) signaling pathways. These findings demonstrate that miR-144 and miR-217 play regulatory roles in inflammatory responses by modulating the NOD1-induced NF-κB signaling pathway.


Inversion of the cerebral mantle as a complication of ventricular shunting. Case report.
The authors report a case of inversion of part of the cerebral mantle into the frontal horn of the lateral ventricle following ventriculoperitoneal shunting for noncommunicating hydrocephalus. The abnormally positioned cerebral mantle continued to grow and caused no detectable neurological dysfunction.


Miniaturization of scorpion beta-toxins uncovers a putative ancestral surface of interaction with voltage-gated sodium channels.
The bioactive surface of scorpion beta-toxins that interact with receptor site-4 at voltage-gated sodium channels is constituted of residues of the conserved betaalphabetabeta core and the C-tail. In an attempt to evaluate the extent by which residues of the toxin core contribute to bioactivity, the anti-insect and anti-mammalian beta-toxins Bj-xtrIT and Css4 were truncated at their N and C termini, resulting in miniature peptides composed essentially of the core secondary structure motives. The truncated beta-toxins (DeltaDeltaBj-xtrIT and DeltaDeltaCss4) were non-toxic and did not compete with the parental toxins on binding at receptor site-4. Surprisingly, DeltaDeltaBj-xtrIT and DeltaDeltaCss4 were capable of modulating in an allosteric manner the binding and effects of site-3 scorpion alpha-toxins in a way reminiscent of that of brevetoxins, which bind at receptor site-5. While reducing the binding and effect of the scorpion alpha-toxin Lqh2 at mammalian sodium channels, they enhanced the binding and effect of LqhalphaIT at insect sodium channels. Co-application of DeltaDeltaBj-xtrIT or DeltaDeltaCss4 with brevetoxin abolished the brevetoxin effect, although they did not compete in binding. These results denote a novel surface at DeltaDeltaBj-xtrIT and DeltaDeltaCss4 capable of interaction with sodium channels at a site other than sites 3, 4, or 5, which prior to the truncation was masked by the bioactive surface that interacts with receptor site-4. The disclosure of this hidden surface at both beta-toxins may be viewed as an exercise in "reverse evolution," providing a clue as to their evolution from a smaller ancestor of similar scaffold.


Adsorption of tributyltin by tributyltin resistant marine Pseudoalteromonas sp. cells.
The isolate, Pesudoalteromonas sp. TBT1, could grow to overcome the toxicity of tributyltin chloride (TBTCl) up to 30 microM in the absence of Cl(-) in the medium until the cells reached an exponential phase of growth. The viability, however, was reduced after the cells reached a stationary phase. The degradation products, such as dibutyltin (DBT) and monobutyltin (MBT), were not detected in the growth medium, indicating that the isolate has no ability to degrade TBT into less toxic DBT and MBT. Up to about 10(7.5) TBT molecules were adsorbed by a single cell. The observation of morphological changes with an electron microscope showed that the cell surface became wrinkled after exposure to the lethal concentration of 10 mM TBTCl. These results indicate that the resistance of the isolate toward the toxicity of TBTCl is not related to the unique cell surface, which seems to play an important role in preventing the diffusion of TBTCl into the cytoplasm.


Sampling effort/frequency necessary to infer individual acute stress responses from fecal analysis in Greylag geese (Anser anser).
Measuring hormone metabolites from excreta is a powerful method to study hormone-behavior relationships. Currently, fecal corticosterone metabolite concentrations are used to estimate individual short-term stress responses. From the free-roaming, semitame flock of greylag geese (Anser anser), as many fecal samples as possible were collected over 3 h following a challenge (social density stress) or in a control situation. This time span corresponds to the gut passage time of geese. It was asked how many samples were necessary to determine differences in excreted corticosterone immunoreactive metabolites (CORTs) between control and social density stress and which parameters (means, maxima, range) reliably showed this difference. A large variation of CORT was found between consecutive samples. Still, means, maxima, and ranges of the samples in a fecal series consistently showed the response to a stressor both within and between individuals. Three samples sufficed if the maximum value of CORT was used, whereas four or more samples were necessary to work with the mean. It was concluded that by increasing the number of fecal samples collected, the course of CORT could be measured more precisely and an individual's acute stress response inferred more reliably.


Nanostructured Lipid Carriers- A Versatile Carrier for Oral Delivery of Lipophilic Drugs.
Around 40% of newly discovered chemical entities in pharmaceutical industries have poor water solubility and hence they suffer from low oral bioavailability owing to undesirable physicochemical and pharmacokinetic properties. So, it is the challenge for the formulation scientists to develop the oral formulation that can mitigate the pitfalls associated with such lipophilic drugs. Lipid nanoparticles hold a promising tool to decrease the pitfalls of lipophilic drugs as lipid components can effectively increase the absorption of drugs, which leads to improvement in oral bioavailability. They are also considered as safe because they are made up of physiological lipids, which are biocompatible and biodegradable in nature. Amongst the lipid nanoparticles, Nanostructured Lipid Carriers (NLCs) are the second-generation lipid nanoparticles and were developed to conquer the limitations of solid lipid nanoparticles. They increase the solubility, permeability, reduce metabolism, P-glycoprotein efflux, and thereby increase the bioavailability of poorly soluble drugs. This review highlights the various aspects of NLCs, such as structural components, types, in vivo fate, pharmacokinetic, toxicity, recent applications, and patent reviews of NLCs in drug delivery.


Separation of two forms of cytochrome P450 adrenal cortex mitochondria.
The separation of two distinct forms of cytochrome P450 from adrenal cortex mitochondria has been achieved by the following steps; (1) lyophilisation (2) iso-octane extraction, (3) (NH(4))(2)SO(4) fractionation in the presence of sodium cholate. The fraction precipitating between 25-35 percent (NH(4))(2)SO(4) gave a difference spectrum with 11-deoxycorticosterone (11-DOC) but not with 20alpha-hydroxycholesterol (20alpha-HOC). This fraction showed high 11beta-hydroxylase activity but low activity for side chain cleavage of cholesterol (S.C.C.). The fraction precipitating between 45-60 percent (NH(4))(2)SO(4) gave a difference spectrum with 20alpha-HOC but not with 11-DOC and exhibited high S.C.C. activity but low 11beta-hydroxylase activity. The absorption spectrum of the 45-60 percent fraction indicated a preponderance of high spin hemoprotein (lambda(max) 395 nm).


Occipitocervical fusion in an infant with atlantooccipital dislocation.
For children who survive traumatic atlantooccipital dislocation (AOD), early surgical stabilization and arthrodesis of the occipitoatlantoaxial complex is typically performed. Because of the unique and crowded anatomy of the occipitocervical junction, the creation of a fusion construct that is both safe and biomechanically sound is extremely challenging, especially in infants. We present a technical report of a patient with infantile type I AOD with gross instability, who underwent surgical stabilization consisting of occiput to C2 arthrodesis using autologous rib, augmented with bone morphogenic protein 2 (BMP-2), Mersilene suture, and Ethibond suture as "cross-connectors." The patient is a 12-month-old female infant who was involved in a high-speed motor vehicle accident and sustained a type I AOD. Definitive surgical stabilization consisting of arthrodesis from the occiput to C2 using autologous rib augmented with BMP-2, Mersilene suture, and Ethibond suture as cross-connectors was performed. There were no intraoperative complications. A follow-up cervical spine computed tomography obtained 12 weeks postoperatively demonstrated excellent occipitocervical alignment, with evidence of modest bony fusion from the occiput to C2. Neurologic examination demonstrated modest, but progressive, improvement, with partial resolution of bilateral sixth nerve palsies, and improvement in upper- and lower-extremity muscle strength and tone. Significant surgical challenges exist regarding occipitocervical fusion in infants with AOD and gross instability. This case report illustrates the successful application of BMP-2-augmented occipitocervical fusion using autologous rib, in combination with Mersilene and Ethibond suture, in the treatment of an infant with type I AOD with gross instability.


Evaluation of microcrystalline chitosans for gastro-retentive drug delivery.
In vivo absorption studies were carried out in human volunteers to evaluate whether microcrystalline chitosan (MCCh) granules would be gastro-retentive. Furosemide, which is site-specifically absorbed from the upper gastrointestinal tract, was used as model drug. The rate of release of furosemide in vitro could be prolonged by increasing the molecular weight (M(w)) or amount of MCCh (150 to 240 kDa; 80 to 95%) in the granules, and also by addition of acidic excipients to the formulations. No marked changes in the in vivo absorption rate (t(max)) were noted, but the amounts of furosemide absorbed (AUC(0- infinity ) and C(max)) decreased as the in vitro release rate decreased, although this was not statistically significant in the case of AUC. The highest AUC(0- infinity ) (3050 micro g l(-1) h) for furosemide (40 mg) was achieved with granules containing 80% MCCh of 150 kDa M(w). With MCCh of 240 kDa M(w) AUC(0- infinity ) was 1890 micro g l(-1) h. This kind of pharmacokinetic profile of furosemide suggests that the gastric retention time of the granules is too short in relation to the release rate, and a large amount of the drug passes its "absorption window" before being released. The in vivo study produced no evidence that the chitosan formulations studied can be used as mucoadhesive gastro-retentive drug delivery systems. The results of in vitro mucoadhesion studies did not predict the results of in vivo studies.


Primary Sclerosing Cholangitis: What the Gastroenterologist and Hepatologist Needs to Know.
Primary sclerosing cholangitis (PSC) is a chronic, idiopathic biliary tract disease characterized by segmental strictures. The disease is progressive with no proven treatments and may eventually lead to cirrhosis and end-stage liver disease. Abrupt changes in liver biochemistries, pain, and/or cholangitis may suggest a dominant stricture amenable to endoscopic therapy or the development of cholangiocarcinoma. Patients with PSC are at increased risk of cholangiocarcinoma. There is a strong association with inflammatory bowel disease, and an associated increased risk of colorectal cancer. Colonoscopy every 1 to 2 years is appropriate.


Further evidence of an association between adolescent bipolar disorder with smoking and substance use disorders: a controlled study.
Although previous work suggests that juvenile onset bipolar disorder increases risk for substance use disorders and cigarette smoking, the literature on the subject is limited. We evaluated the association of risk for substance use disorders and cigarette smoking with bipolar disorder in adolescents in a case-control study of adolescents with bipolar disorder (n=105, age 13.6+/-2.5 years [mean]; 70% male) and without bipolar disorder ("controls"; n=98, age 13.7+/-2.1 years; 60% male). Rates of substance use and other disorders were assessed with structured interviews (KSADS-E for subjects younger than 18, SCID for 18-year-old subjects). Bipolar disorder was associated with a significant age-adjusted risk for any substance use disorder (hazard ratio[95% confidence interval]=8.68[3.02 25.0], chi(2)=16.06, p<0.001, df=1), alcohol abuse (7.66 [2.20 26.7], chi(2)=10.2, p=0.001, df=1), drug abuse (18.5 [2.46 139.10], chi(2)=8.03, p=0.005, df=1) and dependence (12.1 [1.54 95.50], chi(2)=5.61, p=0.02, df=1), and cigarette smoking (12.3 [2.83 53.69], chi(2)=11.2, p<0.001, df=1), independently of attention deficit/hyperactivity disorder, multiple anxiety, and conduct disorder (CD). The primary predictor of substance use disorders in bipolar youth was older age (BPD-SUD versus BPD+SUD, logistic regression: chi(2)=89.37, p<0.001). Adolescent bipolar disorder is a significant risk factor for substance use disorders and cigarette smoking, independent of psychiatric comorbidity. Clinicians should carefully screen adolescents with bipolar disorder for substance and cigarette use.


Secreted frizzled-related protein 5: A promising therapeutic target for metabolic diseases via regulation of Wnt signaling.
Metabolic diseases pose a significant global health challenge, characterized by an imbalance in metabolism and resulting in various complications. Secreted frizzled-related protein 5 (SFRP5), an adipokine known for its anti-inflammatory properties, has gained attention as a promising therapeutic target for metabolic diseases. SFRP5 acts as a key regulator in the Wnt signaling pathway, exerting its influence on critical cellular functions including proliferation, differentiation, and migration. Its significance extends to the realm of adipose tissue biology, where it plays a central role in regulating inflammation, insulin resistance, adipogenesis, lipid metabolism, glucose homeostasis, and energy balance. By inhibiting Wnt signaling, SFRP5 facilitates adipocyte growth, promotes lipid accumulation, and contributes to a decrease in oxidative metabolism. Lifestyle interventions and pharmacological treatments have shown promise in increasing SFRP5 levels and protecting against metabolic abnormalities. SFRP5 is a pivotal player in metabolic diseases and presents itself as a promising therapeutic target. An overview of SFRP5 and its involvement in metabolic disorders and metabolism is provided in this comprehensive review. By elucidating these aspects, valuable insights can be gained to foster the development of effective strategies in combating metabolic diseases.


Inhibitory effects of the anthocyanins from Lycium ruthenicum Murray on angiotensin-I-converting enzyme: in vitro and molecular docking studies.
Lycium ruthenicum Murray (LRM), a perennial shrub plant belonging to the Solanaceae family, is rich in anthocyanins, which have anti-inflammatory, antioxidant, lipid-lowering, intestinal flora regulating, and other pharmacological qualities. This study was primarily aimed to investigate the inhibitory effect of different anthocyanin purities from LRM on angiotensin-I-converting enzyme (ACE) activity in vitro. Moreover, the inhibitory mechanism was further analyzed by molecular docking technology. Two main anthocyanin isomers were identified by ultra-performance liquid chromatography-tandem mass spectrometry and proton/carbon-13 nuclear magnetic resonance, namely petunidin-3-O-[rhamnopyranosyl-(trans-p-coumaroyl)]-5-O-(β-d-glucopyranoside) (trans-Pt3R5G) and petunidin-3-O-[rhamnopyranosyl-(cis-p-coumaroyl)]-5-O-(β-d-glucopyranoside) (cis-Pt3R5G), with a molar ratio of 9:1. Three purification grades of Pt3R5G all showed excellent inhibitory effects on ACE, with the half maximal inhibitory concentration (IC50 ) values being 0.562, 0.421, and 0.106 mg·mL-1 . Increasing the purity may reduce the IC50 within a certain concentration range. An enzymatic kinetic experiment showed that the inhibitory effect of Pt3R5G on ACE was reversible and non-competitive: Pt3R5G and substrate were not in competition for the active sites of ACE. Molecular docking technology further revealed the possible mechanism was that Pt3R5G and ACE amino acid residues were interacting by hydrogen bonds to exert the inhibitory effect. The results indicated that Pt3R5G from LRM was highly effective at inhibiting ACE activity in vitro, with the hydrogen bonds of Pt3R5G and ACE amino acid residues exerting the inhibition. As a potential plant-based ACE inhibitor, Pt3R5G can be used as a functional ingredient for antihypertensive effects. © 2023 Society of Chemical Industry.


Cloning and expression of avian glutamine phosphoribosylpyrophosphate amidotransferase. Conservation of a bacterial propeptide sequence supports a role for posttranslational processing.
The cDNA for glutamine phosphoribosylpyrophosphate amidotransferase, the regulatory enzyme of de novo purine nucleotide biosynthesis, has been cloned for the first time from an animal. The derived amino acid sequence of the avian amidotransferase is homologous with amidotransferase sequences from bacteria and yeast. An 11-amino acid propeptide in Bacillus subtilis amidotransferase is conserved in the avian enzyme. Expression in Chinese hamster ovary (CHO) cells and Escherichia coli provides evidence for two post-translational maturation steps needed for synthesis of active enzyme: incorporation of an iron component and processing of the 11-amino acid propeptide. Functional complementation of a CHO amidotransferase mutant suggests that both maturation steps take place in CHO cells. In contrast, function in E. coli requires deletion of the sequence encoding the propeptide. Defective assembly of the iron component may restrict propeptide removal and activation of the avian amidotransferase in E. coli.


Natural killer cell intrinsic toll-like receptor MyD88 signaling contributes to IL-12-dependent IFN-γ production by mice during infection with Toxoplasma gondii.
Myeloid differentiation factor 88 (MyD88)-dependent IL-12 secretion by dendritic cells is critical for natural killer cell-mediated IFN-γ production and innate resistance to Toxoplasma gondii. Although MyD88(-/-) mice challenged with T. gondii have defective IL-12 responses and succumb to infection, administration of IL-12 to MyD88(-/-) mice fails to prevent acute mortality, suggesting that MyD88 may mediate signals within natural killer cells important for IL-12-dependent IFN-γ production and innate resistance to this parasite. In this study, we found that T. gondii antigens and IL-12 could synergistically trigger IFN-γ secretion by natural killer cells, which was dependent on toll-like receptor-MyD88 signaling. Further analysis showed that p38 mitogen-activated protein kinase, extracellular signal-regulated kinase, c-Jun N-terminal kinase and NF-κB multiple pathways downstream of MyD88 contributed to IFN-γ production by natural killer cells. Moreover, the well-established toll-like receptor agonists, T. gondii profilin (Tgprofilin) and T. gondii heat shock protein 70 (TgHSP70) could evoke a similar IFN-γ secretory response in natural killer cells to that evoked by T. gondii antigens. In vivo adoptive transfer experiments showed that, upon challenge with T. gondii, NOD/SCID-β2 microglobulin null (NOD/SCID-β2m(-/-)) mice injected i.v. with MyD88(-/-) natural killer cells had reduced serum IFN-γ levels and increased splenic tachyzoite burdens compared with those injected i.v. with wild-type natural killer cells. Taken together, these findings demonstrate a critical role for natural killer cell intrinsic toll-like receptor-MyD88 signaling in IL-12-dependent early IFN-γ production and innate resistance to T. gondii.


Genome sequencing and population genomics modeling provide insights into the local adaptation of weeping forsythia.
Understanding the genetic basis underlying the local adaptation of nonmodel species is a fundamental goal in evolutionary biology. In this study, we explored the genetic mechanisms of the local adaptation of Forsythia suspensa using genome sequence and population genomics data obtained from specific-locus amplified fragment sequencing. We assembled a high-quality reference genome of weeping forsythia (Scaffold N50 = 7.3 Mb) using ultralong Nanopore reads. Then, genome-wide comparative analysis was performed for 15 natural populations of weeping forsythia across its current distribution range. Our results revealed that candidate genes associated with local adaptation are functionally correlated with solar radiation, temperature and water variables across heterogeneous environmental scenarios. In particular, solar radiation during the period of fruit development and seed drying after ripening, cold, and drought significantly contributed to the adaptive differentiation of F. suspensa. Natural selection exerted by environmental factors contributed substantially to the population genetic structure of F. suspensa. Our results supported the hypothesis that adaptive differentiation should be highly pronounced in the genes involved in signal crosstalk between different environmental variables. Our population genomics study of F. suspensa provides insights into the fundamental genetic mechanisms of the local adaptation of plant species to climatic gradients.


[Evaluation of the efficacy of an influenza vaccine campaign among health personnel for the prevention of influenza-like illness (ILI)].
Aim of the study is to evaluate influenza vaccination effectiveness in health care workers. Results show: (i) a significant decrease of ILI-cases among vaccinated health care workers, (ii) a significant decrease of sickness absence among vaccinated workers, (iii) a favourable cost/benefit analysis.


Evaluation of the value of plasma concentration of copeptin in the first prenatal visit to diagnose gestational diabetes mellitus.
Gestational diabetes mellitus (GDM) is increasing worldwide. The aim of this study was to investigate whether copeptin concentrations, measured at the first prenatal visit, are associated with risk of GDM. From July 2015 to June 2016, consecutive women who admitted to the obstetrics center of our hospital were included. Data for fasting plasma glucose and copeptin concentrations at the first prenatal visit and one-step GDM screening with 75-g oral glucose tolerance test performed between 24 and 28 weeks of gestation were collected and analyzed. The relationship between levels of copeptin and GDM were evaluated using univariate and multivariate regression analysis. In this study, 101 out of the 827 women developed GDM (12.2%). The GDM distribution across the copeptin quartiles ranged between 4.4% (first quartile) and 25.1% (fourth quartile). For each 1 log-unit increase in plasma concentration of copeptin, the unadjusted and adjusted risk of GDM increased by 1442% (odds ratio 15.42 [95% CI 3.35-54.25], P < 0.001) and 642% (7.42 [2.69-16.02], P < 0.001), respectively. In a multivariate model using the fourth quartiles of copeptin versus quartiles 1 through 3 together with the clinical variables, the marker displayed prognostic information (GDM: OR for fourth quartile, 3.11 [95% CI 1.95-5.24; P = 0.001]). The net reclassification improvement statistic showed that the addition of copeptin to established risk factors significantly increased the correct reclassification of GDM (P = 0.02). The integrated discrimination improvement statistic found that the copeptin level significantly increased discrimination between women with GDM and without GDM (P = 0.01). High copeptin concentrations at the first prenatal visit were associated with increased risk of GDM and might be useful in identifying women at risk of GDM for early prevention strategies.


The establishment of rat hybridoma cell lines secreting McAb against strains of potato virus Y and analysis of its stability.
The rat splenocytes immunized with potato virus Y (PVYn) and ratmyeloma (IR983) were fused by PEG (M. W.1450). Three kinds of stable hybridoma cell lines secreting specific monoclonal antibodies (McAbs) were derived. One kind of the cell lines producing McAbs reacts to PVYn specifically. Another reacts to PVYo specifically. The third one reacts to both of the two strains. Tested by the methods of sandwich-ELISA and indirect-ELISA, all kinds of McAbs did not react to seven plant viruses: tobacco mosaic (TMV), cucumber mosaic (CMV), tobacco tech (TEV), alfalfa mosaic (AMV), turnip mosaic (TuMV), potato leaf roll (PLRV), potato virus X (PVX). The biological properties of the hybridoma cell lines and the McAbs were tested.


The Role of Salvage Resection After Definitive Radiation Therapy for Non-small Cell Lung Cancer.
This study evaluated outcomes of patients who undergo extended delay to resection after definitive radiation therapy for non-small cell lung cancer (NSCLC). Perioperative outcomes and 5-year overall survival of patients with NSCLC who underwent definitive radiation therapy, followed by resection, from 2004 to 2020 in the National Cancer Database were evaluated. Patients who underwent resection >180 days after the initiation of radiation therapy (including any external beam therapy at a total dose of >60 Gy) were included in the analysis. Subgroup analyses were conducted by operation type and pathologic nodal status. From 2004 to 2020, 293 patients had an extended delay to resection after definitive radiation therapy. The clinical stage distribution was stage I to II in 53 patients (18.1%), stage IIIA in 111 (37.9%), stage IIIB in 106 (36.2%), stage IIIC in 13 (4.4%), and stage IV in 10 (3.4%). Median dose of radiation therapy received was 64.8 Gy (interquartile range, 60.0-66.6 Gy). Median days from radiation therapy to resection were 221.0 (interquartile range, 193.0-287.0) days. Lobectomy (64.5%) was the most common operation, followed by pneumonectomy (17.1%) and wedge resection (7.5%). For wedge resection, lobectomy, and pneumonectomy, the 30-day readmission rate was 4.8%, 4.8%, and 8.3%, the 30-day mortality rate was 0%, 3.4%, and 6.4%, and the 90-day mortality rate was 0%, 6.2%, and 12.8%, respectively. Overall survival at 5 years for patients with pN0, pN1, and pN2 disease was 38.6% (95% CI, 30.0-47.2), 43.3% (95% CI, 16.3-67.9), and 24.0% (95% CI, 9.8-41.7), respectively. In this national analysis, extended delay to resection after definitive radiation therapy was associated with acceptable perioperative outcomes among a highly selected patient cohort.


Differential expression of sphingolipids in P-glycoprotein or multidrug resistance-related protein 1 expressing human neuroblastoma cell lines.
The sphingolipid composition and multidrug resistance status of three human neuroblastoma cell lines were established. SK-N-FI cells displayed high expression and functional (efflux) activity of P-glycoprotein, while multidrug resistance-related protein 1 was relatively abundant and most active in SK-N-AS cells. These two cell lines exhibited higher sphingolipid levels, compared to SK-N-DZ, which had the lowest activity of either ATP-binding cassette transporter protein. SK-N-DZ cells also differed in ganglioside composition with predominant expression of b-series gangliosides. In conclusion, these three neuroblastoma cell lines offer a good model system to study sphingolipid metabolism in relation to ATP-binding cassette transporter protein function.


A kinetic and thermodynamic study on hydrolysis of sodium laurate in aqueous phase accompanied by transfer into oil phases containing different organic additives (I).
The kinetic and thermodynamic behavior at the interface between an aqueous solution of sodium laurate (NaLA) and various oil phases comprised primarily of benzene (Bz) and/or different organic compounds including amphiphiles has been investigated in regard to the hydrolysis of NaLA accelerated at the interface, transfer of lauric acid (LA) into oil phase and reverse transfer of Bz into aqueous phase in addition to interface tension. The contact of aqueous NaLA solution with the oil phase was found to accompany the mass transfer of LA and simultaneously promote the hydrolysis of NaLA in water phase. Analysis of the change of OH- ion concentration ([OH-]) over time allowed us to treat the events as a first order reaction. From the rate constant data the activation parameters such as the activation enthalpy and entropy, both of which control the transfer of LA molecules, were determined. The parameters were found to depend greatly on varied situations of the oil phase, being clearly able to explain the physicochemical behavior of the interface. Comparing the cases where the oil phase is one of the respective single systems such as Bz, dodecane (C12) and dodecylbenzene (C12Bz), C12Bz resulted in the lowest rate constant. The transfer (or hydrolysis) rate was measured for the amphiphile-added oil systems as a function of amphiphile concentration. When 0.206 M C16OH-Bz came in contact with aqueous phase, emulsion formation at the interface layer was brought about with approximately zero activation enthalpy, leading to facile or spontaneous transfer of LA. In addition, UV absorbance representing the transfer of Bz from the oil phase to the aqueous phase also demonstrated the effects of added amphiphiles on the action of the interface.


Green detection of trace cyanuric acid and free chlorine together via ion chromatography.
Chlorinated cyanurates (CCAs) are a type of disinfectants currently used worldwide for fight of Coronavirus. However, CCAs upon dosed into water can release not only free chlorine (FC), a strong disinfectant, but also cyanurate (CYA), a persistent compound potentially harmful to human and environment. Therefore, detecting CYA and FC in water are very important not only for ensuring sufficient disinfection but also for monitoring the impacts of FC and CYA on receiving watershed. However, conventional analytical methods for them are mostly based on colorimetric methods, which have high method detection limits (MDLs) and rely on chemical reactions that are likely sensitive to coexisting chemicals. To overcome these issues, we herein proposed a facile and reaction-free method to detect CYA and FC together in just one run by ion chromatography (IC) equipped with both conductivity and ultraviolet absorbance detectors. The method features obvious advantages over colorimetric methods in being lower MDLs (3.6 μg/L for CYA and 9.0 μg/L for FC), environmental-friendly (i.e., no organic solvent involved), and more resistant to alkaline solution. With this method, trace levels of CYA (i.e., 34-44 μg/L), which were nondetectable by conventional method, were found in two river water samples, implying that the local environment was already polluted by CCAs during the pandemic period. Overall, this study demonstrates a robust tool that may assist better understanding and monitoring the fate and transport of trace CCA derivatives in water.


The haemocyanin of the whelk, Busycon contrarium (Conrad): aggregation states and subunit structure.
1. The haemocyanin of the left-handed whelk Busycon contrarium (Conrad) exists largely as six or more multi-decameric aggregates characterized by sedimentation coefficients of approximately 105S, 132S, 155S, 170S, 185S and about 200-220S. 2. These aggregates represent di- to hepta- or octa-decameric assemblies of the basic haemocyanin decamer having a mol. wt of 4.3 x 10(6)-4.5 x 10(6). 3. The fully dissociated subunits in 8.0 M urea (pH 8.5) and at pH 11.1, 0.01 M EDTA have mol. wts of 4.78 x 10(5) and 4.62 x 10(5), close to one-tenth of the mol. wt of the basic decameric unit of most gastropod haemocyanins. 4. The pH dependence of the mol. wts (Mw), studied by light-scattering at the constant protein concentration of 0.010%, exhibit bell-shaped pH transition profiles with mol. wt values of about 16 x 10(6) in the presence of 0.01 M Mg2+, in the pH region from about pH 4.5-8.0; in the absence of stabilizing divalent ions the observed mol. wt is about 10 x 10(6) at pH 4.5-7.0. Below pH 4.5 and above 7.0-8.0 there is a sharp drop in mol. wt to about 4 x 10(5)-4.5 x 10(5). 5. The transition profiles observed with both the urea and salt series of probes investigated at concentration = 0.010% are found to produce aggregation at low reagent concentrations with mol. wt changes from about 9 x 10(6)-12 x 10(6)-14 x 10(6), followed by a decrease in mol. wt below 4.3 x 10(6)-4.5 x 10(6) of the haemocyanin decamers.(ABSTRACT TRUNCATED AT 250 WORDS)


Penile vascular evaluation of men with Peyronie's disease.
A penile vascular evaluation was performed upon 95 consecutive men with Peyronie's disease (19 potent and 76 impotent) and 100 consecutive impotent men without Peyronie's disease to gain further understanding of the pathophysiology of erectile dysfunction in patients with Peyronie's disease. Only 1 of the 19 potent patients with Peyronie's disease had abnormal arterial blood flow by duplex ultrasonography and none had evidence of veno-occlusive dysfunction by either duplex ultrasonography or dynamic infusion cavernosometry/cavernosography. In contrast, 36% of the impotent men with Peyronie's disease had abnormal arterial blood flow and 59% had evidence of veno-occlusive dysfunction. There was no significant difference in historical risk factors for impotence between the impotent patients with Peyronie's disease and the control population of impotent patients. However, the presence of veno-occlusive dysfunction in the control population (16%) was significantly less than that of the impotent Peyronie's disease patients (p < 0.001). Although patients with Peyronie's disease may suffer impotence due to arterial occlusive disease, our results indicate that the principal cause of impotence in this patient population is veno-occlusive dysfunction.