Characterization of the Agrobacterium octopine-cucumopine catabolic plasmid pAtAg67.
In addition to tumor-inducing agrobacteria, non-pathogenic strains are often isolated from crown gall tumors. Such non-pathogenic strains sometimes contain catabolic plasmids that allow them to take advantage of the opines produced in the tumors. Here we characterize for the first time an octopine catabolic plasmid, pAtAg67, which is derived from an Agrobacterium strain isolated from a grapevine tumor in Crete. By sequence analysis, we deduce that pAtAg67 enables its host to catabolize not only octopine, but also cucumopine and agrocinopine-like compounds. We found that a highly similar set of catabolic genes was present in the Ti plasmids of tumorigenic octopine-cucumopine grapevine strains such as pTiAg57. However, the catabolic genes in octopine-cucumopine Ti plasmids were interrupted by a T-DNA segment. As no T-DNA remnants, virulence genes or border repeats were found in pAtAg67, catabolic plasmid pAtAg67 does not appear to be a degenerate octopine-cucumopine Ti plasmid. In line, plasmid pAtAg67 was found to be compatible with incRh1 octopine Ti plasmids, but to be incompatible with the incRh2 agropine Ti plasmid pTiBo542, forming cointegrates by recombination in the homologous trb genes.


Calcium-phosphate mediated DNA transfer into HEK-293 cells in suspension: control of physicochemical parameters allows transfection in stirred media. Transfection and protein expression in mammalian cells.
This is the first report of transient transfection of suspended cells with purified plasmid DNA in bioreactors or spinner flasks. DNA/calcium phosphate complexes were pumped or injected directly into stirred cultures of the immortalized human embryo kidney cell line 293 (HEK-293) which had been adapted to growth in suspension. We identified culture conditions suitable for this approach and modified the protocol for the generation of precipitate complexes, based on our earlier work. In order to stabilize the 'DNA-vehicle'-complex in the culture medium, we identified pH ranges and ion-concentrations which prevent dissolution or aggregation of the precipitate particles. Such conditions maintained suspended fine particles in spinners or bioreactors for up to 6 hr. During that period, cells and precipitate complexes interacted sufficiently to allow DNA transfer and subsequent expression of recombinant protein. In a simple 5 day batch process, with a starting density of 0.3 × 10(6) cells mL(-1), about 0.5 mg L(-1) of a recombinant tissue plasminogen activator variant was observed.


[Soft tissue manifestations of early rheumatic disease: imaging with MRI].
The aim of this study was to evaluate typical magnetic resonance imaging (MRI) findings in early rheumatic diseases manifesting at the soft tissues of the hand using a retrospective analysis. A total of 186 MRI examinations of patients with clinical suspicion of a rheumatic disease were evaluated in a consensus reading by two experienced radiologists. All imaging patterns were assessed with respect to their type and localization. Under blinded and non-blinded conditions diagnoses were correlated with final clinical diagnosis. The most frequent diagnoses were rheumatoid arthritis (RA, 45.7%) and psoriatic arthritis (PsA, 15.6%). The mean correlation between clinical and MRI diagnosis (r) was 0.75 in blinded and 0.853 in non-blinded reading (p <0.001). The following extra-articular imaging patterns were found: synovitis (59.1%), tendovaginitis (91.4%), dactylitis (14.5%), and bone marrow edema (18.3%). Only dactylitis was specific for a particular rheumatic disease (PsA; r=0.934; sensitivity 84.9%, specificity 82.4%). Inflammatory conditions of the hand can be reliably detected with MRI. In many cases the definite diagnosis can only be made when taking clinical, serological, and radiographic results into account (+13.7% increase of significance).


Nine years' experience with the Björk-Shiley prosthetic valve: early and late results of 932 valve replacements.
Between November, 1970, and December, 1977, 932 consecutive patients received the standard Björk-Shiley prosthesis. Operative mortality was 4.6% for aortic valve replacement (AVR; N = 364), 5.1% for mitral valve replacement (MVR; N = 313); 11.3% for multiple valve replacement (N = 194), and 13.7% for valve replacement combined with coronary artery bypass grafting (CABG; N = 51). Factors influencing hospital mortality included type of valve replacement, age at operation, whether the valve replacement was done as an emergency, and year of implantation. Complete follow-up (mean, 3.7 years) was achieved in 95% of the survivors. Actuarial survival was 82% for AVR patients at 8 years, 83% for MVR patients at 7 years, 72% for multiple valve replacement patients at 7 years, and 76% for those who had valve replacement with CABG at 3 years. No significant difference in late survival was found between patients with mitral insufficiency alone (p greater than 0.2) or aortic insufficiency alone (p greater than 0.9) and those with stenotic lesions only. Thromboembolic complications occurred at an incidence of 1.17% per patient-year, and Dicumarol-induced hemorrhages occurred at an incidence of 2.11% per patient-year. Twelve out of 13 patients survived operation for prosthetic valve thrombosis, indicating that this complication is not as catastrophic as many believe. The incidence of reoperation in anticoagulated patients was 0.63% per patient-year. All of these results are compared with data from the recent literature.


Development of a recombinant cell-based indirect immunofluorescence assay (RC-IFA) for the determination of autoantibodies against "rings and rods"-associated inosine-5'-monophosphate dehydrogenase 2 in viral hepatitis C.
Autoantibodies against so-called "rings and rods" structures, as determined by indirect immunofluorescence assay (IFA) using the human cell line HEp-2, have been described in chronic hepatitis C virus (HCV) infected patients treated with interferon/ribavirin. Recently, cytidine triphosphate synthase (CTPS) and inosine-5'-monophosphate dehydrogenase 2 (IMPDH2), the enzyme inhibited by ribavirin, were proposed as the target antigens. We wanted to confirm the identification and setup a robust system for autoantibody testing in routine laboratories. CTPS and IMPDH2 were individually expressed in HEK293 cells and the recombinant cells were used in IFA (RC-IFA) to analyze sera from 33 anti-"rings and rods" antibody positive individuals with unknown diagnosis, 50 patients with chronic HCV infection, 100 with autoimmune hepatitis, 50 with primary biliary cirrhosis and 50 healthy blood donors. We found that all sera with anti-"rings and rods" reacted with recombinant IMPDH2 but none with CTPS. In western blot or ELISA, anti-IMPDH2 positive sera reacted only weakly, if at all, with Escherichia coli derived recombinant IMPDH2 indicating that the autoantibody reaction probably depends on the 3-dimensional conformation of the antigen. A rabbit hyperimmune serum raised against bacterially expressed IMPDH2 produced the ring/rods pattern in IFA using HEp-2. We conclude, that IMPDH2 is indeed the main target of anti-"rings and rods" while CTPS is an unlikely target. Moreover, the novel RC-IFA test system allows a standardized semi-quantitative determination of anti-IMPDH2 basing on a defined recombinant antigen.


Assessment of hindgut function in premature newborns.
Anorectal manometry was performed on 32 small for dates and or preterm infants, using a micro-tip pressure transducer. A normal rectoanal reflex was observed in otherwise normal infants on the first day of life. During the neonatal period, hindgut function in these infants was studied longitudinally using the minimum excitation energy (MER) to elicit a rectoanal reflex. Data suggest that hindgut dysfunction (indicated by a significant increase in MER) develops prior to the onset of abdominal distension or necrotizing enterocolitis (NEC), rather than being present from birth. Consequently, this abnormality of function probably does not initiate the process that leads to NEC. This increase in MER may be a warning that the infant is about to develop abdominal distension or NEC.


Robotic right hemicolectomy: Analysis of 108 consecutive procedures and multidimensional assessment of the learning curve.
Surgeons tend to view the robotic right colectomy (RRC) as an ideal beginning procedure to gain proficiency in robotic general and colorectal surgery. Nevertheless, oncological RRC, especially if performed with intracorporeal ileocolic anastomosis confectioning, cannot be considered a technically easier procedure. The aim of this study was to assess the learning curve of the RRC performed for oncological purposes and to evaluate its safety and efficacy investigating the perioperative and pathology outcomes in the different learning phases. Data on a consecutive series of 108 patients undergoing RRC with intracorporeal anastomosis between June 2011 and September 2015 at our institution were prospectively collected to evaluate surgical and short-term oncological outcomes. CUSUM (Cumulative Sum) and Risk-Adjusted (RA) CUSUM analysis were performed in order to perform a multidimensional assessment of the learning curve for the RRC surgical procedure. Intraoperative, postoperative and pathological outcomes were compared among the learning curve phases. Based on the CUSUM and RA-CUSUM analyses, the learning curve for RRC could be divided into 3 different phases: phase 1, the initial learning period (1st-44th case); phase 2, the consolidation period (45th-90th case); and phase 3, the mastery period (91th-108th case). Operation time, conversion to open surgery rate and the number of harvested lymph nodes significantly improve through the three learning phases. The learning curve for oncological RRC with intracorporeal anastomosis is composed of 3 phases. Our data indicate that the performance of RRC is safe from an oncological point of view in all of the three phases of the learning curve. However, the technical skills necessary to significantly reduce operative time, conversion to open surgery rate and to significantly improve the number of harvested lymph nodes were achieved after 44 procedures. These data suggest that it might be prudent to start the RRC learning curve by treating only benign diseases and to reserve the performance of oncological resection to when at least the initial learning phase has been completed.


Infantile abdominoscrotal hydrocele: a not so benign condition.
Infantile abdominoscrotal hydrocele is a rarely described condition. We report the outcome of the largest single institution experience managing these lesions. We retrospectively reviewed descriptive case series of all infants treated between January 1998 and December 2007. Postoperative followup ranged from 3 to 107 months (median 48). A total of 18 consecutive patients underwent abdominoscrotal hydrocele repair at age 3 to 21 months (median 8). The first 13 patients underwent inguinal hydrocelectomy, which consisted of high ligation of the processus vaginalis and complete excision of the abdominal component of the lesion. The remaining 5 patients underwent plication of the tunica vaginalis accomplished through the scrotum. Included in this group were 2 patients initially treated expectantly because the lesion was without undue tension at diagnosis. Both cases demonstrated spontaneous resolution of the abdominal component of the lesion but ultimately required surgery for scrotal hydrocele. Overall 23 testes (5 bilateral lesions) were operated on, of which 18 had some degree of testicular dysmorphism detected by ultrasonography or during surgery and 15 fully recovered postoperatively. A total of 11 patients experienced 1 or more postoperative complications, including persistent scrotal swelling (7), hypoplastic testis (3), scrotal hematoma (2), inguinal hernia (1) and cryptorchidism (1). Morbidity related to scrotal hydrocelectomy was negligible. There were no recurrences of abdominoscrotal hydrocele in the series. Tense infantile abdominoscrotal hydroceles are associated with a high rate of testicular dysmorphism, which is often reversed by early intervention. Simple transscrotal plication of the tunica vaginalis represents an effective procedure with decreased postoperative complications.


Factors associated with admission of children to an intensive care unit and readmission to hospital within 28 days of discharge: A population-based study.
Hospital readmissions within 28 days are an important performance measurement of quality and safety of health care. The aims of this study were to examine the rates, trends and characteristics of paediatric intensive care unit admissions, and factors associated with readmissions to hospital within 28 days of discharge. This retrospective, population-based record linkage study included all children ≥28 days and <16 years old admitted to an intensive care unit (ICU) in a New South Wales (NSW) public hospital from 2004 to 2013. Data were sourced from the NSW Admitted Patients Data Collection and the NSW Registry of Births, Deaths and Marriages, Death Registration. We identified 21 200 ICU admissions involving 17 130 children. Admissions increased by 24% over the study period with the greatest increase attributed to respiratory and musculoskeletal conditions. A higher proportion of children were <5 years, male, lived in major cities, were publicly insured and had chronic conditions. The median length of ICU stay was 42 h and overall hospital stay was 7 days. There were 905 deaths, two-thirds during the index admission with the leading causes being injuries, cancer and infections. Twenty-three per cent of ICU admissions were readmitted to hospital within 28 days of discharge. Associated independent factors were younger age, longer index hospital stay and emergency index admission. Children with chronic conditions of cancer and genitourinary disorders were more likely to be readmitted. Identification of complex chronic conditions, consideration of long-term health planning and interventions intended to reduce readmission is warranted in order to reduce the burden to families and the health-care system.


Calbindin-D28k is a more reliable marker of human Purkinje cells than standard Nissl stains: a stereological experiment.
In a study of human Purkinje cell (PC) number, a striking mismatch between the number of PCs observed with the Nissl stain and the number of PCs immunopositive for calbindin-D28k (CB) was identified in 2 of the 10 brains examined. In the remaining eight brains this mismatch was not observed. Further, in these eight brains, analysis of CB immunostained sections counterstained with the Nissl stain revealed that more than 99% Nissl stained PCs were also immunopositive for CB. In contrast, in the two discordant brains, only 10-20% of CB immunopositive PCs were also identified with the Nissl stain. Although this finding was unexpected, a historical survey of the literature revealed that Spielmeyer [Spielmeyer W. Histopathologie des nervensystems. Julius Springer: Berlin; 1922. p. 56-79] described human cases with PCs that lacked the expected Nissl staining intensity, an important historical finding and critical issue when studying postmortem human brains. The reason for this failure in Nissl staining is not entirely clear, but it may result from premortem circumstances since it is not accounted for by postmortem delay or processing variables. Regardless of the exact cause, these observations suggest that Nissl staining may not be a reliable marker for PCs and that CB is an excellent alternative marker.


The anti-tumour effect of cisplatin and ifosfamide on xenografted squamous cell carcinoma of the head and neck is schedule-dependent.
The role of chemotherapy (CHX) in squamous cell carcinoma of the head and neck (SCCHN) has been expanding. Although combination chemotherapy regimens regularly produce significantly high response rates, meta-analyses show little benefit regarding final outcome. One way to improve induction CHX is to improve drug combinations and schedules for CHX. Cisplatin (CDDP) is one of the most active drugs in the treatment of patients with SCCHN, and it is used in most combinations. Ifosfamide (IFO) is another agent that has shown activity in the treatment of patients with SCCHN. A poorly differentiated squamous cell carcinoma xenografted to nude mice was used. CDDP (2.5 mg/kg) and IFO (100 mg/kg) as single bolus doses induced significant retardation of tumour growth. Single drug administration was compared with CDDP given before IFO and IFO given before CDDP. Mean specific growth delay (SGD) for untreated controls was 0. For CDDP as single drug it was 1.50, for IFO as single drug it was 0.79, for CDDP given 4 h before IFO it was 1.79, and for IFO given 4 h before CDDP it was 2.90. Maximum toxicity, calculated as change in median weight at day 7, was less than 10%. The efficacy and toxicity of CDDP and IFO is schedule-dependent, with IFO given before CDDP being more effective than CDDP given before IFO. This is in contrast to most schedules used clinically. The toxicities were comparable. The number of combinations of drugs with respect to order and time interval is of a magnitude that would not be possible to test clinically. In the pursuit of more efficient combinations, the importance of order and schedule of drugs and also the potential of xenografted SSCHN are underestimated.


Thromboplastic and fibrinolytic activities of cultured human cancer cell lines.
Thromboplastic and fibrinolytic activities of 14 lines of cultured human cancer cells were estimated by modified Astrup's methods. High tissue thromboplastic activity was found in one line of urinary-bladder cancer, 2 lines of gastric cancer and one line of lung cancer, but no activity was found in 6 lines of lung cancer. High fibrinolytic activity was noted in one line of gastric cancer and 2 lines of lung cancer, but no activity was seen in 6 lines of lung cancer and one line of gastric cancer. No plasmin activity was found. The tumour cell lines could be classified into 3 groups on the basis of the 2 activities. Cancer cell lines could also be classified into 2 groups: with high or low release of thromboplastin into culture media. Fibrinolytic activity was found in the culture media of all cell lines with high fibrinolytic activity. Fibrinolytic activity, but not thromboplastic activity, seemed to be influenced by the constituents of culture media. No definite correlation was found between the 2 activities and the histological types of the parent tumours of the cultured cells.


Villus growth and cell replacement in the small intestine of the neonatal pig.
Cells produced in the crypts of newborn pig ileum migrate onto villi during development. There is little or no corresponding loss of cells from villus tips during the first week of postnatal life. Villus growth during this period is largely responsible for the slow rate of cellular renewal seen to take place.


Relationships between total plasma load of torquetenovirus (TTV) and TTV genogroups carried.
In 239 torquetenovirus-positive people, multiple-genogroup infections were common and associated with higher viral loads than would be expected from simple additive effects. The latter observation was restricted to the infections which included both genogroups 1 and 3, pointing to the possible existence of some kind of infection facilitation between these genogroups.


Identification of Key Residues Associated with the Interaction between Plutella xylostella Sigma-Class Glutathione S-Transferase and the Inhibitor S-Hexyl Glutathione.
Glutathione S-transferases (GSTs) are important detoxification enzymes involved in the development of metabolic resistance in Plutella xylostella. Uncovering the interactions between representative PxGSTs and the inhibitor S-hexyl glutathione (GTX), helps in the development of effective PxGST inhibitors for resistance management. As the PxGST most severely inhibited by GTX, PxGSTσ (sigma-class PxGST) adopts the canonical fold of insect GSTs. The formation of the PxGSTσ-GTX complex is mainly driven by H-bond and hydrophobic interactions derived from the side chains of favorable residues. Of the residues composing the active site of PxGSTσ, Lys43 and Arg99 are two hot spots, first reported in the binding of GSH derivatives to GSTs. Such differences indicate the metabolism discrimination of different insect GSTs. Unfavorable interactions between the PxGSTσ active site and GTX are depicted as well. The research guides the discovery and optimization of PxGSTσ inhibitors.


[Change in recurrent inhibition when standing in normal subjects and spastics (author's transl)].
In normal subjects and patients with diseases of the upper motor neurone, recurrent inhibition directed to soleus motoneurones was studied in three different postures: sitting (control situation), standing when leaning backward on a wall, standing independently without the support of a wall. In normal subjects, this recurrent inhibition was not modified when standing with the support of a wall, while it was enhanced when standing without such a support.


Monocytic leukemia with cutaneous involvement: ultrastructural observations on unusual cytoplasmic complexes.
A case of pure monocytic leukemia ("true" histiocytic lymphoma) presented as multiple cutaneous nodules. The cells contained unusual cylindric formations of the rough endoplasmic reticulum that differ from classic ribosomal-lamellar complexes.


Prevalence of Theileria in the tick Hyalomma detritum detritum in the Doukkala region, Morocco.
The overall prevalence of Theileria species, mainly, if not exclusively, T.annulata, in 901 Hyalomma detritum detritum collected from cattle in the Doukkala region of Morocco over a period of 2 years was 21.5%. The quantity of infection (number of sporoblasts per infected tick) followed the negative binomial distribution with between one and 250 sporoblasts per infected tick. Infected ticks were found in eight of fourteen areas examined whilst T.annulata was present in all fourteen. There were significant differences in both the prevalence and the quantity of infection between ticks collected from different farms, and between nymphs collected in the autumn from these farms, and moulted in the laboratory, and adults collected in the following summer. The prevalence, but not the quantity, of infection was higher in female than in male ticks. No correlations were established between infection of engorged nymphs and the breed, sex and Theileria piroplasm parasitaemia of the host animal. However, calves infected a greater proportion of nymphs than adult cattle and the heavier the infestation of nymphs on an animal, up to a plateau, the higher the prevalence of infection in those nymphs. There were no differences in infection between ticks moulted at 24 degrees C and 37 degrees C, after the engorged nymphs had been stored at 18 degrees C to simulate over-wintering.


A Pilot Study of High-Definition Transcranial Direct Current Stimulation in Refractory Status Epilepticus: The SURESTEP Trial.
Refractory status epilepticus (RSE) is a life-threatening emergency with high mortality and poor functional outcomes in survivors. Treatment is typically limited to intravenous anesthetic infusions and multiple anti-seizure medications. While ongoing seizures can cause permanent neurological damage, medical therapies also pose severe and life-threatening side effects. We tested the feasibility of using high-definition transcranial direct current stimulation (hd-tDCS) in the treatment of RSE. We conducted 20-min hd-tDCS sessions at an outward field orientation, intensity of 2-mA, 4 + 1 channels, and customized for deployment over the electrographic maximum of epileptiform activity ("spikes") determined by real-time clinical EEG monitoring. There were no adverse events from 32 hd-tDCS sessions in 10 RSE patients. Over steady dosing states of infusions and medications in 29 included sessions, median spike rates/patient fell by 50% during hd-tDCS on both automated (p = 0.0069) and human (p = 0.0277) spike counting. Median spike rates for any given stimulation session also fell by 50% during hd-tDCS on automated spike counting (p = 0.0032). Immediately after hd-tDCS, median spike rates/patient remained down by 25% on human spike counting (p = 0.018). Compared to historical controls, hd-tDCS subjects were successfully discharged from the intensive care unit (ICU) 45.8% more often (p = 0.004). When controls were selected using propensity score matching, the discharge rate advantage improved to 55% (p = 0.002). Customized EEG electrode targeting of hd-tDCS is a safe and non-invasive method of hyperacutely reducing epileptiform activity in RSE. Compared to historical controls, there was evidence of a cumulative chronic clinical response with more hd-tDCS subjects discharged from ICU.


Clinical implications of prospective genomic profiling of metastatic breast cancer patients.
Metastatic breast cancer remains incurable. Next-generation sequencing (NGS) offers the ability to identify actionable genomic alterations in tumours which may then be matched with targeted therapies, but the implementation and utility of this approach is not well defined for patients with metastatic breast cancer. We recruited patients with advanced breast cancer of any subtype for prospective targeted NGS of their most recent tumour samples, using a panel of 108 breast cancer-specific genes. Genes were classified as actionable or non-actionable using the European Society of Medical Oncology Scale for Clinical Actionability of Molecular Targets (ESCAT) guidelines. Between February 2014 and May 2019, 322 patients were enrolled onto the study, with 72% (n = 234) of patients successfully sequenced (n = 357 samples). The majority (74%, n = 171) of sequenced patients were found to carry a potentially actionable alteration, the most common being a PIK3CA mutation. Forty-three percent (n = 74) of patients with actionable alterations were referred for a clinical trial or referred for confirmatory germline testing or had a change in therapy outside of clinical trials. We found alterations in AKT1, BRCA2, CHEK2, ESR1, FGFR1, KMT2C, NCOR1, PIK3CA and TSC2 to be significantly enriched in our metastatic population compared with primary breast cancers. Concordance between primary and metastatic samples for key driver genes (TP53, ERBB2 amplification) was > 75%. Additionally, we found that patients with a higher number of mutations had a significantly worse overall survival. Genomic profiling of patients with metastatic breast cancer can have clinical implications and should be considered in all suitable patients.


Prostaglandins mediate lipopolysaccharide effects upon cholecystokinin and neurotensin phenotypes in neuroendocrine corticotropin-releasing hormone neurons: in situ hybridization evidence in the rat.
Intraperitoneal injection of the endotoxin lipopolysaccharide produces an inflammation accompanied by immune system activation and secretion of cytokines that stimulate the hypothalamo-pituitary-adrenal (HPA) axis to release the anti-inflammatory corticosterone. Upstream in HPA axis are neuroendocrine corticotropin-releasing hormone neurons in the paraventricular nucleus whose multipeptidergic phenotype changes during inflammation: coexisting corticotropin-releasing hormone and cholecystokinin mRNAs are up-regulated whereas neurotensin mRNA expression is induced de novo. These changes may be mediated by prostaglandins released from perivascular and microglial cells in response to circulating cytokines. We examined by quantitative in situ hybridization histochemistry whether blockade of prostaglandin synthesis by indomethacin alters phenotypic expression in paraventricular nucleus neurons after lipopolysaccharide. Because indomethacin also elevated circulating corticosterone, animals were adrenalectomized and corticosterone replaced. Results showed that i.p. indomethacin administration suppressed lipopolysaccharide effects in a phenotype non-specific manner: one injection was sufficient to prevent both the increase in corticotropin-releasing hormone and cholecystokinin mRNAs expression and the induction of neurotensin mRNA expression. Therefore, neuroendocrine corticotropin-releasing hormone neurons with different peptidergic phenotypes appear to respond as a whole in the acute phase response to systemic infection.


Functional connectivity of proximal and distal lower limb muscles and impact on gait variability in stroke.
Higher gait variability after stroke increases risk of falls and compromises safe community ambulation. Corticomotor connectivity plays an important role in walking after stroke, however, its relation to gait variability remains unknown. Do corticomotor characteristics of the proximal and distal lower limb muscles predict gait variability in individuals with chronic stroke? Retrospective analysis of data from 30 individuals with chronic stroke was conducted. Corticomotor characteristics were measured in the paretic and non-paretic tibialis anterior (TA, distal muscle) and rectus femoris (RF, proximal muscle) using transcranial magnetic stimulation. We calculated corticomotor excitability ratio of paretic TA and RF (CMETA/RF), corticomotor excitability symmetry (CMEsym) between hemispheres for the TA and RF, and ipsilateral corticomotor excitability (ICE) of the paretic TA. Gait variability was quantified as the coefficient of variation of the paretic step length (spatial) and step time (temporal) during comfortable walking. Relations between corticomotor characteristics and gait variability were tested with multiple linear regression. CMETA/RF and CMEsym of RF were significant predictors of spatial gait variability. Greater corticomotor input to the paretic RF compared to the paretic TA and greater symmetry of RF were related to higher spatial gait variability. There were no significant predictors of temporal gait variability. Corticomotor inputs to the proximal RF may be important for spatial gait variability, reflecting a compensatory role of RF in walking after stroke. Stroke survivors with relatively greater corticomotor input to the paretic RF may adopt compensatory strategy to enhance propulsion and achieve foot clearance, but it may also increase spatial gait variability, particularly when combined with impaired motor control of the paretic TA. These findings may provide novel rehabilitative targets to decrease gait variability and promote safe ambulation in individuals with stroke.


What is happening outside North America regarding human dirofilariasis?
The etiologic agents of human dirofilariasis in the Old World are Dirofilaria immitis, which cause pulmonary and subcutaneous nodules, and Dirofilaria repens, which cause ocular lesions. Although reports of new cases of dirofilariasis are sporadic in other parts of the world, a considerable amount of information is generated in Europe regarding human dirofilariasis. Most cases have been detected in the Mediterranean countries, Ukraine, and Russia; however, isolated or short series of cases have been reported in the Balkan Republics and central and northern European countries. Seroepidemiologic studies have provided evidence that humans living in endemic areas present rates of infection similar to those of the autochthonous canine populations. Antibodies against endosymbiont Wolbachia bacteria have been demonstrated recently in human Dirofilaria infections. During D. immitis infections, preadult worms and third- and fourth-stage larvae are often destroyed by the host reaction, releasing a considerable amount of Wolbachia, and a Th1-type response against Wolbachia and/or filarial antigens is mounted. On the contrary, infections with D. repens, in which worms frequently remain intact, no Th1-type response has been observed. As humans are resistant hosts, the Th1-response could have a role in the resistance against parasites. The causes for the rise in the incidence of human dirofilariasis as well as the possible application of Wolbachia antigens in the serodiagnosis of human infections are discussed.


Tanshinone IIA Sodium sulfonate regulates antioxidant system, inflammation, and endothelial dysfunction in atherosclerosis by downregulation of CLIC1.
Tanshinone IIA Sodium sulfonate (STS) is clinically used for treating cardiovascular diseases in Traditional Chinese Medicine due to its antioxidation and anti-inflammation activities. Intracellular chloride channel 1 (CLIC1) participates in the regulation of oxidative stress and inflammation. This study investigates whether CLIC1 mediates the cardioprotective effects of STS. STS were used to treat atherosclerosis (AS) induced by feeding Apolipoprotein E-deficient (ApoE-/-) mice with a high-fat, cholesterol-rich diet. In addition, normal and CLIC1-/- human umbilical vein endothelial cells were treated with STS after exposure to H2O2 for 12h. The oxidative status was determined by analyzing reactive oxygen species(ROS) and malondialdehyde (MDA) levels. ELISA, qRT-PCR and Western blot were used to determine the levels of TNF-α, IL-6, ICAM-1 and VCAM-1. CLIC1 cellular localization was examined by immunofluorescence. Chloride ion concentration was detected with chloride ion quenchers (MQAE). STS treatment decreased atherosclerotic lesion area by 3.5 times (P = 0.001) in vivo. Meanwhile, STS reduced MDA production (13.6%, P = 0.008), increased SOD activity (113.6%, P = 0.008), decreased TNF-α (38.6%, P = 0.008) and IL-6 (43.0%, P = 0.03) levels, and downregulated the expression of CLIC1, ICAM-1, and VCAM-1 in the atherosclerotic mice. The dose-dependent anti-oxidative and anti-inflammatory effects of STS were further confirmed in vitro. Furthermore, CLIC1 depletion abolished the STS-mediated decrease of ROS and MDA production in HUVEC cells. Additionally, STS inhibited both CLIC1 membrane translocation and chloride ion concentration. The anti-oxidant, and anti-inflammation properties of STS in preventing AS is mediated by its inhibition of CLIC1 expression and membrane translocation.


Low defensive cardiac reactivity as a physiological correlate of psychopathic fearlessness: Gender differences.
Affective/interpersonal features of psychopathy have been consistently associated with diverse psychophysiological indicators of low threat sensitivity, suggesting an underlying deficit in the reactivity of the brain's defensive motivational system. This study examined the Cardiac Defense Response (CDR) -a complex pattern of heart rate changes in response to an aversive, intense, and unexpected stimulus- and its second accelerative component (A2), as a new physiological indicator of the fearlessness trait component of psychopathy. The differential contribution of dispositional fearlessness, externalizing proneness, and coldheartedness to the CDR pattern elicited during a defense psychophysiological test was examined in a mixed-gender sample of 156 undergraduates (62% women) assessed by the Psychopathic Personality Inventory-Revised (PPI-R). Higher PPI-R Fearless Dominance scores were related to lower heart rate changes throughout the CDR in women, but not in men. Further analyses on scales conforming the fearless dominance factor revealed that the hypothesized reduced A2 was specifically related to higher PPI-R Fearlessness scores only in women. Our findings provide initial evidence for the utility of the A2 to better understand the physiological aspects of fearlessness tendencies and its potential distinct manifestations across genders.


In situ aerobic composting eliminates the toxicity of Ageratina adenophora to maize and converts it into a plant- and soil-friendly organic fertilizer.
Ageratina adenophora has invaded many subtropical and tropical countries and caused tremendous ecological and economic losses. This necessitates a new way to use the debris left after clearing this plant. Therefore, the allelochemicals in fresh and aerobically composted A. adenophora plants (FA and CA, respectively) were compared, and their allelopathy against maize was evaluated. The results showed that CA decreased the allelochemicals (6-hydroxy-5-isopropyl-3,8-dimethyl-4a,5,6,7,8,8a-hexahydronaphthalen-2(1H)-one and 4,7-dimethyl-1-(propan-2-ylidene)-1,4,4a,8a-tetrahydronaphthalene-2,6(1H, 7H)-dione) by over 95% compared to FA. In a seed germination test, CA aqueous solutions improved the seed germination and seedling growth, whereas FA solutions led to opposite results. Chemical fertilizers (CF) plus FA resulted in much lower plant biomass and nutrient uptake than CF in a greenhouse experiment. Compared with CF, CF+CA showed positive effects on maize, soil microbial biomass and diversity and enzyme activities in the field. However, the compositions of the predominant microbes were almost unaffected by the application of CA and CF+CA. These significant findings extended our knowledge regarding the elimination of A. adenophora toxicity against other plants and soil microbes through allelochemical degradation in the composting process. In situ aerobic composting provides a new, simple and economical method to convert A. adenophora into a plant- and soil-friendly organic fertilizer.


Effects on bone mineral density of 12-month goserelin treatment in over 40-year-old women with uterine myomas.
We evaluated the effects on bone mineral density (BMD) of a 12-month treatment with goserelin depot, a gonadotropin-releasing hormone agonist, in a group of women with symptomatic uterine myomas requiring hysterectomy. Sixteen women, mean age 45.6 +/- 5.0, reporting menorrhagia associated with uterine myomas, candidates for hysterectomy, were scheduled to be treated with goserelin depot for 12 months. BMD was measured at the vertebral (L2-L4) and proximal femur level (femoral neck and trochanter) at the start of therapy and 6, 12, and 18 months later using dual energy X-ray absorptiometry (Hologic QDR 1000/W). The patients were followed for a minimum of 6 months after the end of treatment. Thirteen of the 16 women enrolled completed the treatment and three suspended it after 5, 6, and 7 months, respectively, because of side effects (hot flashes, insomnia, depression). Of the 13 women who completed the treatment, three underwent hysterectomy because of myoma regrowth and the recurrence of symptoms 3-18 months later; four reached the menopause 5-16 months later, and six were all menstruating normally with a follow-up varying from 6 to 18 months. After 12 months of therapy we observed a bone loss at vertebral, femoral neck, and trochanter of 4.4% (P < 0.05 versus baseline; P = not significant versus 6 months), 7.5% (P < 0.01 versus baseline, P < 0.01 versus 6 months), and 7.6% (P < 0.001 versus baseline, P < 0.05 versus 6 months), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Power to the people.
ICROSS is a small but effective organisation tackling issues by working with the resources and capacities of poor communities.


Effect of biomimetic shear stress on intracellular uptake and cell-killing efficiency of doxorubicin in a free and liposomal formulation.
Shear stress could be considered in the context of cellular uptake and cell-killing efficiency. Thus, mimicking the dynamic characteristics of in vivo environment is important in targeted drug delivery. We investigated the intracellular uptake and cell-killing efficiency of doxorubicin (DOX) in a free and liposomal form (Doxil(®)) under biomimetic shear stress to mimic in vivo environment. In this dynamic environment, cells demonstrated significantly higher fluorescence intensity than that of the static environment, and fluorescence microscopy images indicated increased intracellular uptake of DOX in the presence of fluidic shear stress. In cells treated with free DOX and liposomal Doxil(®), cell-killing efficiency was affected by shear stress. Taken together, shear stress, affecting drug uptake and cell-killing efficiency, is important in intracellular drug targeting.


Downregulating PKC delta provides a PI3K/Akt-independent survival signal that overcomes apoptotic signals generated by c-Src overexpression.
3Y1 rat fibroblasts overexpressing the tyrosine kinase c-Src (3Y1(c-Src) cells) become transformed by downregulation of protein kinase C delta (PKC delta). However, when 3Y1(c-Src) cells were subjected to serum withdrawal, they underwent apoptosis via a cytochrome c/caspase 9 pathway. In contrast, neither parental nor v-Src-transformed 3Y1 cells underwent apoptosis when subjected to serum withdrawal. If PKC delta was downregulated, the apoptotic phenotypes induced by serum withdrawal in the 3Y1(c-Src) cells were suppressed. The apparent survival signal generated by PKC delta downregulation was independent of the phosphatidylinositol-3-kinase (PI3K)/Akt survival pathway. Collectively, these data indicate that (1) c-Src overexpression renders cells sensitive to apoptotic stress, and (2) that downregulation of PKC delta provides a novel PI3K/Akt-independent survival signal capable of suppressing apoptotic signals.


Testosterone Replacement Therapy and the Cardiovascular System.
As testosterone replacement therapy (TRT) has emerged as a commonly prescribed therapy for symptomatic low testosterone, conflicting data have been reported in terms of both its efficacy and potential adverse outcomes. One of the most controversial associations has been that of TRT and cardiovascular morbidity and mortality. This review briefly provides background on the history of TRT, the indications for TRT, and the data behind TRT for symptomatic low testosterone. It then specifically delves into the rather limited data for cardiovascular outcomes of those with low endogenous testosterone and those who receive TRT. The available body of literature strongly suggests that more work, by way of clinical trials, needs to be done to better understand the impact of testosterone and TRT on the cardiovascular system.


Development of an expedient intramolecular Pauson-Khand reaction approach to stereoselectively construct the trans-decalin with a C1 quaternary chiral center.
Stereoselective synthesis of the trans-decalin subunit with a defined C1 quaternary chiral center has been achieved by the Pauson-Khand reaction (PKR) as a key step. The developed chemistry offers an alternative to the IMDA reaction that has been used for the syntheses of trans-decalin based biologically active natural products.


Exploring the antimicrobial, antiviral, antioxidant, and antitumor potentials of marine Streptomyces tunisiensis W4MT573222 pigment isolated from Abu-Qir sediments, Egypt.
Due to the therapeutic importance of microbial pigments, these pigments are receiving the attention of researchers. In this present study 60 isolates were isolated from sediments of Abu-Qir coast of the Mediterranean sea, Alexandria, Egypt, out of which 12 were considered as pigmented actinomycetes. Streptomyces sp. W4 was characterized by small round green pigmented colonies when grown on starch-casein agar medium. The green pigment was extracted using a mixture of acetone-methanol (7:3 v/v). The antimicrobial, antioxidant, antiviral, and anticancer activities of the green pigment produced by Streptomyces sp.W4 were investigated. The pigment was characterized using FTIR, Raman spectroscopy, EDX and GC-MS. The results revealed that the pigment has antibacterial and antifungal activity and also showed inhibition of HAV 78% but its antiviral activity against the Adenovirus was weak. The results proved the safety of the pigment toward normal cells and anticancer activity against three different cancer cell lines HepG-2 (liver cancer cell line), A549 (lung cancer cell line), and PAN1 (pancreas cancer cell line). The pigment was combined with 9 antibiotics and then tested against the Gram-negative bacterium Enterococcus faecalis using disc diffusion bioassay. LEV showed an antagonistic effect, while CXM and CIP showed a synergistic effect.


Response to vWF-coated beads and soluble p-selectin to characterize apheresis platelets: a comparison of three cell separators.
Plateletpheresis technologies differ among various cell separators. Differences in centrifugation force, centrifugation time, and platelet concentration in the platelet concentrate may affect platelet activation and function. In a three-way crossover design, 12 donors were randomly assigned to three types of cell separators, two continuous flow systems (Amicus DN, Fenwall Division, Baxter and Spectra LRS, Gambro BCT) and one intermittent flow system (MCS+, Haemonetics). The response to vWF-coated beads was determined in the peripheral blood and fresh platelet concentrates to obtain information about the initiating step of platelet aggregation, that is, platelet adhesion. Levels of soluble p-selectin were measured as a marker of platelet activation. Platelet yield and concentration of platelets were higher in platelet concentrates obtained with the Amicus DN than with the Spectra LRS or the MCS+. Maximal aggregation was significantly higher in platelets from the Amicus DN than in platelets from the Spectra LRS or the MCS+. Higher concentrations of soluble p-selectin were seen in platelet concentrates obtained with the Amicus DN than in concentrates from the Spectra LRS or the MCS+, but they did not differ after correction for the number of platelets per component. Different plateletpheresis procedures induced distinct changes of platelet function. Platelets collected using the Amicus DN retained the strongest adhesion capacity irrespective of their activation.


Induction of resistance, enzyme activity, and phytochemicals in canola plants treated with abscisic acid elevated based on nutrient availability: a case study on Brevicoryne brassicae L. (Hemiptera: Aphididae).
The cabbage aphid, Brevicoryne brassicae L. (Hemiptera: Aphididae), is one of the important pests of cruciferous plants throughout the world including Iran. In the present study, we grew cultivated canola plants under different fertilizers or distilled water and sprayed them with 100 µM abscisic acid (ABA) or a control solution (NaOH dissolved in water) to study (i) the antibiosis parameters of B. brassicae on these plants; (ii) the antixenosis of B. brassicae adults on these plants; (iii) the plant's peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) activity; and (iv) the plant's total phenolic and glucosinolate content. The results of antibiosis experiments showed that ABA and fertilizers have a profound and negative effect on the performance of B. brassicae. In the antixenosis experiment, control plants attracted a significantly higher number of adult females in comparison to treated plants. Also, B. brassicae had lower performance and preference when they were reared on the ABA-treated fertilized plants with higher levels of phenolic and glucosinolate content. These results prompted us to hypothesize that fertilizers enable canola plants to trigger a higher level of secondary metabolites. Our findings reveal that the type and level of nutrient availability may have different impacts on how the plant regulates its defense mechanisms.


Predictive toxicogenomics in preclinical discovery.
The failure of drug candidates during clinical trials due to toxicity, especially hepatotoxicity, is an important and continuing problem in the pharmaceutical industry. This chapter explores new predictive toxicogenomics approaches to better understand the hepatotoxic potential of human drug candidates and to assess their toxicity earlier in the drug development process. The underlying data consisted of two commercial knowledgebases that employed a hybrid experimental design in which human drug-toxicity information was extracted from the literature, dichotomized, and merged with rat-based gene expression measures (primary isolated hepatocytes and whole liver). Toxicity classification rules were built using a stochastic gradient boosting machine learner, with classification error estimated using a modified bootstrap estimate of true error. Several types of clustering methods were also applied, based on sets of compounds and genes. Robust classification rules were constructed for both in vitro (hepatocytes) and in vivo (liver) data, based on a high-dose, 24-h design. There appeared to be little overlap between the two classifiers, at least in terms of their gene lists. Robust classifiers could not be fitted when earlier time points and/or low-dose data were included, indicating that experimental design is important for these systems. Our results suggest development of a compound screening assay based on these toxicity classifiers appears feasible, with classifier operating characteristics used to tune a screen for a specific implementation within preclinical testing paradigms.


Effects of valinomycin doping on the electrical and structural properties of planar lipid bilayers supported on polyelectrolyte multilayers.
Supported Lipid Bilayers (SLBs) on Polyelectrolyte Multilayers (PEMs) have large potential as models for developing sensor devices. SLBs can be designed with receptors and channels, which benefit from the biological environment of the lipid layers, to create a sensing interface for ions and biomarkers. PEMs assembled by the Layer-by-Layer (LBL) technique and used as supports for a lipid bilayer enable an easy integration of the bilayer on almost any surface and device. For electrochemical sensors, LBL assembly enables nanoscale tunable separation of the lipid bilayer from the electrode surface, avoiding undesired effects of the electrode surface on the lipid bilayers. We study the fabrication of valinomycin-doped SLBs on PEMs as a model system for biophysical studies and for selective ion sensing. SLBs are fabricated from dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylserine (DOPS) 50:50 vesicles doped with valinomycin, as a K+-selective carrier. SLBs were deposited on electrodes coated with poly(allyl amine hydrochloride) (PAH) and poly(styrene sodium sulfonate) (PSS) multilayers. Lipid bilayer formation was monitored by using Quartz Crystal Microbalance with Dissipation (QCMD) technique and Atomic Force Microscopy (AFM). Electrochemical impedance spectroscopy (EIS) and potentiometric measurements were performed to assess K+ selectivity over other ions and the potential of valinomycin-doped SLBs for K+-sensing.


HIV and urethritis: time required for antiretroviral therapy to suppress HIV in semen.
To examine the time required to suppress HIV in the genital tract with antiretroviral therapy (ART) in men with urethritis. An observational cohort study. Men with HIV and urethritis not on ART were enrolled at an STI clinic in Malawi and offered to initiate ART. Blood and semen samples were collected pretreatment and at 1, 2, 4, 8, 12 and 24 weeks posturethritis treatment. Median viral loads (VLs) were calculated by ART initiation groups: 'within 1 week', 'between 1 and 4 weeks' and 'no ART before 4 weeks', based on the men's choice about whether or not to initiate ART. The presence of ART at each visit was confirmed by bioanalytical methods. Between January 2017 and November 2018, 74 men presented with urethritis and HIV and were confirmed ART naive. The median age was 32 years. Forty-one (55% of men) initiated ART within 1 week; 12 (16%) between 1 and 4 weeks; and 21 (28%) did not initiate ART by week 4. Within the 1 week group, median VL was suppressed within 4 weeks in both semen and blood. Among the 1-4 weeks group, VL was suppressed within 4 weeks in semen and 5 weeks in blood. Among the no ART before 4 weeks group, VL in semen declined within the first 4 weeks but remained unsuppressed through week 24, and there was no significant decline in blood HIV. Treatment of urethritis and prompt initiation of ART with counseling for safer sex for at least one month is a critical measure to reduce transmission of HIV.


Synthesis, anti-ferroptosis, anti-quorum sensing, antibacterial and DNA interaction studies of chromene-hydrazone derivatives.
Nineteen chromene-hydrazone derivatives containing a variety of structural modifications on the hydrazone moiety were synthesized. Structure-activity correlations were investigated to determine the influence of structural variations on anti-ferroptosis, anti-quorum sensing, antibacterial, DNA cleavage and DNA binding properties. Ferroptosis inhibitory activity was determined by measuring the ability of the derivatives to reverse erastin-induced ferroptosis. Several of the derivatives were more effective than fisetin at inhibiting ferroptosis, with the thiosemicarbazone derivative being the most effective. Quorum sensing inhibition was evaluated using Vibrio harveyi, and both V. harveyi and Staphylococcus aureus were used to determine antibacterial activity. The semicarbazone and benzensulfonyl hydrazone derivatives showed moderate quorum sensing inhibition with IC50 values of 27 μM and 22 μM, respectively, while a few aryl hydrazone and pyridyl hydrazone derivatives showed bacterial growth inhibition, with MIC values ranging from 3.9 to 125 μM. In addition, the interaction of the hydrazone derivatives with DNA was investigated by gel electrophoresis, UV-Vis spectroscopy and molecular docking. All of the derivatives cleaved plasmid DNA and showed favorable interaction with B-DNA through minor groove binding. Overall, this work highlights a broad range of pharmacological applications for chromene-hydrazone derivatives.


Rapid determination of polyploidy in human chorionic tissue sections.
Chromosomal analysis from aborted tissue has become an important diagnostic aid. However, the necessary cultures are frequently unsuccessful due to the condition of the aborted tissue. Polyploidy, in particular triploidy, in the conceptus is a common cause of early pregnancy loss and unlike aneuploidy does not appear to be associated with an increased recurrence risk. The necessity to monitor a subsequent pregnancy with amniocentesis is therefore eliminated. Therefore, in cases where a chromosomal anomaly is probable, a fast simple method of identification of a polyploid karyotype would be valuable. In this presentation, we describe a method using a scanning light microscope and histologic tissue preparations. This method can accurately determine the ploidy of the aborted material in 5 days.


Childhood allergies and asthma: New insights on environmental exposures and local immunity at the lung barrier.
While certain bacteria and respiratory viruses promote local inflammation and disease onset, a more diverse colonization of the different species in the (gut) microbiome may be linked to more regulatory responses and protection against asthma and allergies. These processes are also influenced in part by food intake, both targeting the composition of the gut microbiome and influencing the immune system via metabolites. Early life environmental microbial exposure also contributes to protection against asthma and allergy and is linked with an early activation of the innate immune system and the development of regulatory immune responses. Although greater mechanistic insight is needed, it is tempting to speculate that part of the environmental effect can be explained by modulation of the microbiome composition at mucosal surfaces, epithelial barrier function and/or local immunity. A review of the latest studies is provided.


Epidemiological Screening for the Risk of Mental, Behavioral and Developmental Disorders, Including Autism, in Early Childhood: Data for Russia 2017-2019.
We aimed to screen children aged 18-48 months in the general population of nine Russian regions for risk of mental, behavioral and developmental disorders (MBDDs) including autism spectrum disorders (ASD) using an original screening tool. The prevalence of the risk for MBDDs is 1307:10,000 (13.07%), the prevalence of clinically verified cases of MBDDs is 151:10,000 (1.51%), whereas the prevalence of ASD among them is 18:10,000 (0.18%). Basing on our results, the screening procedures are already integrated into the Russian primary care system since the end of 2019. Screening of the risk for MBDDs including ASD in Russia among children in the general pediatric population is a promising area of preventive medicine.


Normative Beliefs and Aggression: The Mediating Roles of Empathy and Anger.
This study examined a two-mediator model with both empathy and anger as mediators in the association between children's normative beliefs about aggression and forms (relational and physical) and functions (reactive and proactive) of aggressive behavior. Ninety-eight children (54% males, Mage=46.21months, SD = 8.84months) reported their approval of relationally and physically aggressive behaviors depicted in iconic (animation) and enactive (toy figurines) hypothetical scenarios. Children's aggression, empathy and anger were measured using teacher reports. No main effects of normative beliefs about aggression on the corresponding aggressive behavior were found. Normative beliefs about aggression were negatively associated with empathy and empathy was significantly associated with relational aggression, suggesting that developing social emotional processes mediate the relation between social cognitions and aggression. Anger was associated with aggression, but not normative beliefs about aggression. The findings provide support for the distinction between subtypes of aggressive behavior in young children and the developing social-cognitive and affective processes that influence these behaviors.


Constitutive and barbital-induced expression of the Cyp6a2 allele of a high producer strain of CYP6A2 in the genetic background of a low producer strain.
The levels of one or more cytochrome P450 (CYP) enzymes and the respective mRNAs are found to be higher in insecticide-resistant insects than in susceptible insects. To understand better how insects regulate the levels of CYPs, we examined the expression of the Cyp6a2 gene in various strains of Drosophila melanogaster. We also took a transgenic approach to understand the molecular mechanisms that are involved in strain variation of Cyp6a2 expression. RNA blot analysis showed that the constitutive expression of Cyp6a2 varies from strain to strain; the level of CYP6A2 mRNA is barely detectable in the underproducer ry506 strain, whereas it is very high in the overproducer 91-R and MHIII-D23 strains. The long terminal repeat (LTR) of mobile element 17.6 that is found in the 3' untranslated region (UTR) of the Cyp6a2 gene of some strains does not appear to have any role on the steady-state CYP6A2 mRNA level. We also found that the Cyp6a2 gene is inducible by barbital in 91-R, ry506 as well as 91-C, which carries an LTR insertion. To examine the genetic background of the underproducer ry506 strain with respect to Cyp6a2 expression, we transformed the ry506 strain with the Cyp6a2 allele of the overproducer 91-R strain (Cyp6a2-91 R) and measured the constitutive and barbital-induced expression of the Cyp6a2-91 R transgene in the transformed flies. The Cyp6a2-91 R transgene carrying 129 bp of DNA upstream of the ATG codon did not show any constitutive or barbital-induced expression in the ry506 host genome. However, transgenes with 1331 and 985 bp upstream DNA showed similar levels of constitutive expression that were higher than that of the endogenous Cyp6a2 gene of the ry506 host strain, but lower than the expression of the same gene in the 91-R strain. Both these transgenes, with 1331 and 985 bp upstream DNA, also showed induction with 0.1 M barbital. DNA sequence analysis revealed that in both 91-R and ry506, the upstream DNA between +1 and -985 bp contains a distal and a proximal group of three potential barbie boxes, i.e. cis-elements that are thought to be involved in barbiturate-mediated induction of CYP genes. Except for four bases located near the distal cluster of barbie boxes and two other bases, the base sequence of the upstream DNA is identical in ry506 and 91-R strains. These results suggest that the underproducer ry506 strain has the trans-regulatory factors to support constitutive and induced expression of the Cyp6a2-91 R allele carrying DNA between -129 and -1331 bp regions. Possible reasons for low constitutive expression of the endogenous Cyp6a2 gene and moderate level of expression of the Cyp6a2-91 R allele in the ry506 genetic background are discussed.


[Use of a rhomboid flap to repair temporal and frontotemporal cutaneous defects: 11 cases].
Temporal and frontotemporal skin defects can be repaired using various reconstruction procedures: temporojugal rotation-advancement flaps, frontotemporal advancement flaps, skin grafts, etc. We propose an alternative method using a rhomboid transposition flap, thus extending the possibilities for repair in this region. This is a retrospective, non-comparative study of patients in whom a rhomboid transposition flap of the temporal or frontotemporal region was created between February 2008 and March 2010. Sex, age, hospitalization, histological type of the excised lesion, type of anaesthesia, defect size, possible occurrence of complications and outcome were compiled from the medical records. Eleven patients (five women and six men) of average age 76 years were included. The lesions were due in all cases to basal cell carcinoma. Five patients were ambulatory and seven were hospitalized. All patients underwent surgery under local anaesthesia in a single session. Hospitalized patients had more extensive cutaneous defects, were older and had more comorbidities. One patient presented incomplete lateral resection. The most frequent complication was bruising on the lower eyelid (5/11), and two more severe complications occurred: distal flap necrosis and haematoma. The aesthetic result was good for all patients after a mean 20months of follow-up. Rhomboid transposition flaps appear to be a reconstruction option that warrants consideration in cutaneous defects in the temporal or frontotemporal region.


Knee alignment in the transverse plane during weight-bearing activity and its implication for the tibial rotational alignment in total knee arthroplasty.
The alignment of the knee in the transverse plane is important to the biomechanical functions of the lower limb, and is also associated with the outcomes of the total knee arthroplasty. This study aimed to evaluate the dynamic tibiofemoral alignment in the transverse plane during a weight-bearing activity. Knee kinematics of weight-bearing flexion from 0° to 120° was obtained in 16 healthy subjects utilizing biplanar radiography and 3D-2D registration techniques. The anteroposterior axes of the femur at multiple flexion angles in the range of knee motion were used to calculate the anteroposterior axis of motion using the least square method. The latter was compared to six surface-derived anatomical axes in the proximal tibia which were most commonly reported in the literature. The relationship between the anteroposterior axis of motion and the tibial tubercle was also quantified. The anteroposterior axis of motion did not coincide with any of the six anatomical axes tested. Their orientations varied from 10.3° of external rotation to 9.9° of internal rotation in relation to the former. The anteroposterior axis of motion tended to intersect the tibial tubercle between the medial border and the medial 1/3 point. Instead of any of the tested anatomical axes, the transverse knee alignment during functional movements is associated with the medial 1/3 portion of the tibial tubercle. This region may be helpful for the rotational alignment of the tibial components in total knee arthroplasty.


Demonstration of Ga-67 localization in human placenta.
Gallium-67 uptake was noted in the mid-pelvic uterine area of a 19-week-pregnant female. Lateral and oblique Ga-67 scans revealed an anterior location of radioactivity that correlated with the position of the placenta as determined by ultrasonic technique.


Effect of different doses of almorexant on learning and memory in 8-month-old APP/PS1 (AD) mice.
To explore the effects of different doses of almorexant (an dual orexin receptor antagonist) on learning and memory in Alzheimer's disease (AD) mice. Forty-four APP/PS1 (model of Alzheimer's disease; AD) mice were randomly divided into 4 groups: the control group (CON) and those that received 10 mg/kg almorexant (low dose; LOW), 30 mg/kg almorexant (medium dose; MED) and 60 mg/kg almorexant (high dose; HIGH). During the 28-day intervention period, mice received an intraperitoneal injection at the beginning of the light period (6:00 am). The effects of different doses of almorexant on learning and memory and 24-hour sleep-wake behaviour were assessed by immunohistochemical staining. The above continuous variables are expressed as the mean ± standard deviation (SD), and then univariate regression analysis and generalized estimating equations were performed to compare the groups; these results are expressed as the mean difference (MD) and 95% confidence interval (CI). The statistical software used STATA 17.0 MP. Forty-one mice completed the experiment (3 died: 2 mice in the HIGH group and 1 mouse in the CON group). Compared with the CON group, the LOW group (MD=6803 s, 95% CI: 4470 to 9137 s), MED group (MD=14,473 s, 95% CI: 12,140-16,806 s) and the HIGH group (MD=24,505 s, 95% CI: 22,052-26,959 s) had significantly longer sleep durations. The Y maze results showed that LOW group (MD=0.14,95%CI: 0.078-0.20) and MED group (MD=0.14,95%CI = 0.074-0.20) mice compared to the CON group, and the low-medium dose of Almorexant did not damage the short-term learning and memory performance of APP / PS1 (AD) mice.Compared with the CON, LOW, and MED groups, the HIGH group exhibited a significant decrease in the Aβ plaque-positive area in the cortex (MD= -0.030, 95% CI: -0.035 to -0.025; MD=-0.049, 95% CI: -0.054 to -0.044; and MD=-0.07, 95% CI: -0.076 to -0.066, respectively). The moderate dose of almorexant (30 mg/kg) prolonged the sleep duration of APP/PS1 (AD) mice to a greater extent than the low dose (10 mg/kg) without altering learning and memory. The MED mice showed a good sleep response and a small residual effect on the next day. High-dose (60 mg / kg) almorexant impaired behavioral learning and memory performance in mice.Compared to the CON group and the LOW group, the MED group exhibited improved working memory. Thus, treatment with almorexant may reduce β-amyloid deposition in AD, slowing neurodegeneration. Additional studies are needed to determine the mechanism of action.


Influence of Er:YAG laser surface treatment on flexural and bond strengths to glass-infiltrated zirconia-reinforced ceramic.
To evaluate the effect of Er:YAG laser conditioning of a glass-infiltrated alumina-based zirconia-reinforced ceramic on its flexural strength and on bonding to a resin cement. Sixteen blocks (5 × 5 × 4 mm) and 50 discs (Ø 12 mm, 1 mm thickness) of In-Ceram Zirconia (ICZ) obtained from CAD-CAM blocks were infiltrated with glass. For the microtensile bond strength (μTBS) test, all blocks were treated with aluminum oxide (AOX) and divided into 4 groups (n = 4): G1 (AOX), no combined surface treatment; G2 (ROC), tribochemical silica-coating; G3 (EY200), Er:YAG laser 200 mJ/15 Hz; and G4 (EY250), Er:YAG laser 250 mJ/10 Hz. The ceramic blocks were silanated and cemented with a resin cement (Panavia F2.0/Kuraray) to composite resin blocks and subjected to the μTBS test. For the flexural strength evaluation, the discs were divided into 5 groups (n = 10) as described above, in addition to a control group (G5 - CTRL, mirror-polished without further treatment). Each surface treatment was submitted to qualitative evaluation under SEM. One-way ANOVA (α = 5%) revealed the highest bond strength value for EY200 with no significant difference from ROC. The groups AOX and EY250 showed similar μTBS values that were statistically lower than those of EY200. For flexural strength, ROC was the only group with significantly lower values when compared with the CTRL. The use of Er:YAG laser at 200 mJ/15 Hz can be considered an innovative and effective alternative for surface conditioning of ICZ since it did not reduce the flexural strength of the ceramic and improved the resin cement bond to this substrate.


Escalator-related injuries among older adults in the United States, 1991-2005.
This study describes the epidemiology of escalator-related injuries among adults age 65 and older in the U.S. between 1991 and 2005, through a retrospective analysis of data from the National Electronic Injury Surveillance System of the U.S. Consumer Product Safety Commission. There were an estimated 39,850 escalator-related injuries and no fatalities. The overall injury rate was 7.8/100,000 population. During the study period 1991-2005, the rate of escalator-related injuries doubled. The mean age of the study population was 80.1 years (S.D.=8.5 years) with 73.3% female. The most frequent cause of injury was a slip, trip or fall (84.9%, 95% CI: 82.7-87.2%). The most frequently injured body parts were the lower extremities (25.9%, 95% CI: 21.5-30.2%) and the head (25.0%, 95% CI: 20.5-29.5%). The leading type of injury was soft tissue injuries (54.2%, 95% CI: 49.7-58.7%) followed by lacerations (22.3%, 95% CI: 18.4-26.1%) and fractures (15.6%, 95% CI: 13.1-18.1%). The rate of head injuries and the rate of hospitalizations increased with age. Escalator-related injuries occur infrequently but may result in significant trauma. These injuries are often associated with a slip, trip or fall. Awareness of the risks and the circumstances leading to escalator injuries allows for better direction of intervention strategies on the part of injury prevention specialists.


Wnt, GSK3, and Macropinocytosis.
Here we review the regulation of macropinocytosis by Wnt growth factor signaling. Canonical Wnt signaling is normally thought of as a regulator of nuclear β-catenin, but emerging results indicate that there is much more than β-catenin to the Wnt pathway. Macropinocytosis is transiently regulated by EGF-RTK-Ras-PI3K signaling. Recent studies show that Wnt signaling provides for sustained acquisition of nutrients by macropinocytosis. Endocytosis of Wnt-Lrp6-Fz receptor complexes triggers the sequestration of GSK3 and components of the cytosolic destruction complex such as Axin1 inside multivesicular bodies (MVBs) through the action of the ESCRT machinery. Wnt macropinocytosis can be induced both by the transcriptional loop of stabilized β-catenin, and by the inhibition of GSK3 even in the absence of new protein synthesis. The cell is poised for macropinocytosis, and all it requires for triggering of Pak1 and the actin machinery is the inhibition of GSK3. Striking lysosomal acidification, which requires macropinocytosis, is induced by GSK3 chemical inhibitors or Wnt protein. Wnt-induced macropinocytosis requires the ESCRT machinery that forms MVBs. In cancer cells, mutations in the tumor suppressors APC and Axin1 result in extensive macropinocytosis, which can be reversed by restoring wild-type protein. In basal cellular conditions, GSK3 functions to constitutively repress macropinocytosis.


The interaction between human papilloma viruses related cancers and non-coding RNAs.
Human papillomaviruses (HPVs) constitute a number of double-stranded DNA viruses with propensity to cause infection in squamous epithelial cells. Certain types of these viruses have been found to cause human cancers through delivering their oncoproteins E6 and E7. Since not all of infected patients develop malignant lesions, other factors might affect HPV-associate carcinogenic processes. A number of investigations have shown interaction between HPV-encoded proteins and a number of non-coding RNAs, principally microRNAs (miRNAs) and long non-coding RNAs (lncRNAs). Such interactions have been found to influence pathogenesis of HPV-related cancers. miR-21, miR-9, miR-143, miR-214 and let-7 are among miRNAs that contribute in the pathogenesis of HPV-related lesions. HOTAIR, SNHG8, SOX2OT, SNHG12, GABPB1-AS1, SOX21-AS1, DINO, HOST2, CCDST, FAM83H-AS1, TMPOP2 and CCEPR are examples of lncRNAs that contribute in this process. In the current review, we provide an outline of investigations that reported the impact of these transcripts in HPV-related cancers.


Patients with active infection with Paracoccidioides brasiliensis present a Th2 immune response characterized by high Interleukin-4 and Interleukin-5 production.
Paracoccidiodomycosis (PCM) is caused by the dimorphic fungus Paracoccidioides brasiliensis. In humans, the disease presents a broad spectrum of clinical manifestations, ranging from localized mucocutaneous lesions to a widespread manifestation with involving the mononuclear phagocyte system. In attempt to better understand the regulation of immune response during the infection, this study analyzed the production of regulatory and inflammatory cytokines in 25 infected patients and 19 health controls. Regulatory and inflammatory cytokines were analyzed in supernatants of peripheral blood mononuclear cells (PBMC) stimulated with mitogens or soluble P. brasiliensis antigens. A pattern of Th2 immune response was observed in patients, mainly attributed to a higher production of IL-4 and IL-5 than to a lower production of IFN-gamma. Patients with disseminated infection presented undetectable levels of IFN-gamma after antigen stimulation and high levels of IL-1beta, which were probably associated with the inflammatory reaction observed in multifocal infection.


[Hydatidosis in Sardinia. Co-involved parameters and hypothesis for a control campaign].
Although hydatidosis provokes in man and livestock losses of economic significance, there has been no comprehensive study on this disease. This article is an account of Sardinia as an important area of echinococcosis-hydatidosis. The incidence and distribution of infection in the intermediate and definitive hosts are reviewed and the current status of hydatid disease is evaluated. The review is divided into main sections: (a) geographical and environmental data; (b) domestic intermediate hosts; (c) definitive host; (d) human infections. An average of 85% of the sheep are infected and about 75% of cysts are fertile, 13% of the dogs are parasitized and mainly live as strays, up to 39% of the shepherd's dogs were found to harbour the mature worm of Echinococcus granulosus. Hospital data indicate that even nowadays about 200 people are operated every year with an incidence of 11.1/100.000 inhabitants. The socio-cultural factors (seminomadism, slaughtering practice, etc.) involved in the dissemination, the economic implication of this disease, the benefits due to an eradication, the feasibility of a campaign and the organizational structure aimed at eradicating echinococcosis/hydatidosis in this area, are discussed.


Theophylline clearance during and after mild upper respiratory infection.
Theophylline clearance was estimated by a single-sample technique in 19 subjects with acute clinical respiratory infections. Mean (+/- SD) theophylline clearance during acute illness was 0.044 +/- 0.011 L X h-1 X kg-1 compared to 0.043 +/- 0.012 L X h-1 X kg-1 1 month later. In a subset of five subjects, single-sample clearance estimates were not different from standard multi-sample clearance estimates. Mild upper respiratory infection does not appear to alter theophylline clearance in adults.


Post-menopausal women and osteoporosis: available choices for maintenance of skeletal health.
To briefly summarize the therapeutic choices for osteoporosis prevention which are currently available to post-menopausal women. Results of randomized clinical trials and epidemiological studies in postmenopausal women, and pre-clinical studies in ovariectomized rats were summarized. Estrogen combined with progestogen in hormone replacement therapy (HRT) is effective in relieving perimenopausal symptoms and maintaining bone mineral density. However, the increased breast cancer risk associated with long-term HRT use makes it a less desirable option for many women. Selective estrogen receptor modulators (SERMs), such as raloxifene, are also effective in maintaining bone density, without stimulating the breast or uterus. However, SERMs do not relieve perimenopausal hot flashes. HRT is effective for acute relief of perimenopausal symptoms, but for women who are unwilling or unable to take HRT long-term, SERMs such as raloxifene are a useful therapy for the prevention of osteoporosis.


[Characteristics of cerebral hemodynamics and psychological adaptation of patients with myocardial infarction during period of rehabilitation and convalescence].
Cerebral circulation and psychological state of 93 myocardial infarction patients aged from 29 to 60 years have been studied. The comparison of data concerning cerebral circulation with psychological profile of these patients enables us to make a conclusion that there is interrelation between the state of cerebral hemodynamics which ensures brain homeostasis and psychological adaptation of patients with myocardial infarction at different stages of the disease


Contribution of vascular smooth muscle cell apoptosis to spiral artery remodeling in early human pregnancy.
Uterine spiral artery remodeling is one of the key maternal adaptations of pregnancy, allowing delivery of the large volumes of maternal blood required for both placental and fetal growth. Failure of this process is associated with obstetric complications including preeclampsia, fetal growth restriction and miscarriage. Spiral artery remodeling is characterized by loss of the musculoelastic wall which is replaced by fibrinoid and intramural extravillous trophoblast cells (EVT). In recent years attention has focused on the initial stages of spiral artery remodeling which include separation of the vascular smooth muscle cells (VSMCs) and their phenotypic switch to a more synthetic phenotype, facilitating their migration away from the vessel wall. However, less is known about the final fate of the VSMCs. In vitro studies suggested that EVT could induce VSMC apoptosis, though VSMC apoptosis is not seen within the wall of the spiral arteries undergoing remodeling. However, apoptotic VSMCs have been observed amongst those cells which had migrated away from the vessel wall. This review article gives a brief overview of the current state of knowledge of the processes involved in spiral artery remodeling, and then concentrates on the mechanisms involved in VSMC apoptosis, drawing on knowledge from other vascular beds.


Comparative efficacy evaluation of moxidectin gel and ivermectin paste against internal parasites of equines in Brazil.
A total of 24 male and female equines of mixed breed, 10-20 months of age and naturally infected with internal parasites was utilized in a controlled test to evaluate the efficacy of a moxidectin 2% gel formulation at the dosage of 0.4 mg moxidectin per kg of live weight and an ivermectin 1.87% commercial paste formulation at the dosage 0.2 mg ivermectin per kg applied orally. Animals were allocated into three groups of eight horses each based on pre-treatment eggs per gram (EPG) counts and treatments were randomized among the groups. One group was kept as untreated controls. One animal in the moxidectin-treated group died before the end of the trial from a cause unrelated to treatment leaving a total of seven animals in this group. Fecal egg counts were performed three times post-treatment and the number of parasites remaining in each animal was determined. Statistical analyses using geometric means were performed at the 1% level of significance. Both moxidectin and ivermectin preparations reduced initial EPG from a mean of 1600 to 0 on Days 5, 7 and at the end of the trial on Day 14. Efficacy percentages of moxidectin and ivermectin against immature and adult nematodes were as follows: Trichostrongylus axei, Parascaris equorum, Strongylus edentatus, S. vulgaris, Triodontophorus spp. and Gyalocephalus capitatus, 100% for both products; Habronema muscae 99.5 and 99.6%, respectively, Strongyloides westeri, 100 and 99.2%, respectively; Oxyuris equi, 99.6 and 100%, respectively; small strongyles, 99.7% for both products. Of the latter, the most numerous were: Cylicocyclus insigne, Cylicostephanus longibursatus and Cyathostomum catinatum. No Gasterophilus nasalis were found in horses from either treated group, while two of eight control horses had infections with thisparasite. Moxidectin showed greater efficacy (84.9%) than ivermectin (67.8%) against Strongylus vulgaris larvae found in the mesenteric artery aneurisms, but the difference was not statistically significant. Total parasite counts for both treated groups were significantly lower (p<0.01) than in the non-treated group. No significant differences were noted between moxidectin and ivermectin. Efficacy against the 30 nematode species found in this study was very evident for both products. As expected, neither moxidectin nor ivermectin was effective in controlling the tapeworm Anoplocephala perfoliata. No adverse reactions were observed during the experimental period.


Binding of a rhodamine-labeled lipopolysaccharide to lipopolysaccharide-responder and nonresponder lymphocytes.
The binding levels of a highly soluble rhodamine-labeled lipopolysaccharide (LPS-Rh) on different LPS-responsive and nonresponsive murine spleen cells were measured with a cytofluorometer (CFM). In all the experiments, a preferential binding of LPS-Rh to LPS-responsive cells was observed. Furthermore, only a proportion of B cells from the responder mouse strain was selectively stained after addition of LPS-Rh. These findings suggested that LPS-Rh binds to specific components of the cell membrane, involved in the triggering of B-cell mitogenicity.


A new species of the idiocerine leafhopper genus Chinaocerus Zhang amp; Li (Hemiptera: Cicadellidae) from northwest China, with a checklist and key to species.
A new leafhopper species, Chinaocerus dentatus Zhang sp. nov., is described and illustrated from Qinghai and Gansu Provinces, China. Specimens were collected from willow (Salix sp.). A checklist and key to species of the genus are provided.


"Do-not-touch" lesions of bone revisited.
Incidental bone lesions are encountered frequently in day-to-day practice. Many of these lesions are indeterminate requiring referral to specialist centres for further characterisation with or without biopsy; however, as biopsy has its own drawbacks, not all lesions can be subjected to biopsy. The primary role of a radiologist in these situations is to characterise these lesions based on their imaging appearances into aggressive lesions requiring biopsy and non-aggressive lesions that do not require a biopsy. The term "do-not-touch lesion" is used to describe a lesion with typical radiographic appearances that can be characterised based on radiographic appearances alone without needing a biopsy. With recent advances in imaging, many incidental lesions can be characterised into do-not-touch lesions based on their imaging appearances alone using a single imaging technique or using a combination of imaging techniques and, less frequently, with the additional help of serological investigations, without the need for biopsy. Hence, the definition of do-not-touch lesions of bone needs a revisit. In this article, we attempt to redefine do-not-touch lesions of bone and propose an imaging-based classification for characterisation of these lesions.


Methicillin-resistant Staphylococcus aureus: related infections and antibiotic resistance.
Staphylococcus aureus is a well adapted human pathogen, capable of living freely in the inanimate environment and spreading from person to person, existing as a colonizer or commensal, hiding in intracellular compartments and, most importantly, inducing various forms of human disease. Infections caused by S. aureus, above all by antibiotic-resistant strains, have reached epidemic proportions globally. The overall burden of staphylococcal disease caused by antibiotic-resistant S. aureus, particularly by the methicillin-resistant strains, is increasing in many countries, including Italy, in both healthcare and community settings. The widespread use of antibiotics has undoubtedly accelerated the evolution of S. aureus, which, acquiring multiple resistance genes, has become able to survive almost all antibiotic families; this evolution versus more resistant phenotypes has continued among the newer agents, including linezolid and daptomycin. The diminished clinical usefulness of vancomycin is seen as one of the most worrisome problems in many clinical settings and in many countries. In fact, the increasing spread of heteroresistant vancomycin-intermediate S. aureus (hVISA) and vancomycin intermediate (VISA) strains adds new problems, not only in terms of the treatment of severe infections sustained by these microorganisms, but also in the microbiological definition of susceptibility.


Atherosclerosis: from genetic polymorphisms to system genetics.
It is generally considered that the genetics of atherosclerosis and its complications involves a large number of genes with common alleles having weak effects on disease risk but possibly interacting with each other and with non-genetic factors. In such a complex system, absence of marginal effects (effects of polymorphisms considered one at a time) is insufficient to exclude the implication of a polymorphism on disease risk. Investigating polymorphisms and even genes one by one is no longer appropriate. It is necessary to focus on biological systems and integrate the contribution of genetic as well as non-genetic factors and their interactions. Hopefully, system genetics will ultimately improve our understanding of the genetic architecture of complex traits.


Histopathologic consequences of surgical approaches to the singular nerve.
Selective sectioning of the singular nerve has proven successful in the relief of incapacitating benign positional vertigo (BPV), presumably on the histopathologic basis of posterior ampullary cupulolithiasis. Although the surgical techniques of transmeatal and the recently introduced retrosigmoid IAC approach have been well described, little has been written about the singular nerve's anatomical relationships determined histologically for each surgical approach. The anatomical relationships of the singular nerve in both the transmeatal and retrosigmoid IAC approaches are discussed. Histologic evidence presented suggests that other mechanisms may also be ultimately responsible for improvement in the symptoms of BPV following transmeatal surgery. Moreover, the high incidence of injury to the endolymphatic duct during retrosigmoid surgery raises questions about the exact role of the endolymphatic duct and sac in inner ear homeostasis.


Gallic acid binding to Spatholobus parviflorus lectin provides insight to its quaternary structure forming.
Therapeutic effects of gallic acid (GA) have already been extensively studied. However, its interaction with lectins has not gained much attention. It is of interest to validate the binding profile of GA with Spatholobus parviflorus seed lectin. A combination of Isothermal Titration Calorimetry (ITC), haemagglutination assay and molecular docking was applied on SPL-GA interaction. ITC results showed four binding sites, stoichiometry, n=4, irrespective of the ratio of SPL:GA taken for titration. Difference among the four binding sites of a single molecule of SPL with regard to GA binding kinetic parameters was consistently varying. Similarly, the glide scores obtained for GA in the four different binding clefts of SPL were also conformed to the ITC. The binding of GA on SPL without affecting its sugar binding property could be considered as a boon for glycobiological research. From the presented studies, it could be proposed that the SPL-GA interactions may facilitate drug delivery by specific targeting/attachment by profiling of cell-surface glycans, followed by controlled release of drugs.


Effects of assortative mate choice on the genomic and morphological structure of a hybrid zone between two bird subspecies.
Phenotypic differentiation plays an important role in the formation and maintenance of reproductive barriers. In some cases, variation in a few key aspects of phenotype can promote and maintain divergence; hence, the identification of these traits and their associations with patterns of genomic divergence is crucial for understanding the patterns and processes of population differentiation. We studied hybridization between the alba and personata subspecies of the white wagtail (Motacilla alba), and quantified divergence and introgression of multiple morphological traits and 19,437 SNP loci on a 3,000 km transect. Our goal was to identify traits that may contribute to reproductive barriers and to assess how variation in these traits corresponds to patterns of genome-wide divergence. Variation in only one trait-head plumage patterning-was consistent with reproductive isolation. Transitions in head plumage were steep and occurred over otherwise morphologically and genetically homogeneous populations, whereas cline centres for other traits and genomic ancestry were displaced over 100 km from the head cline. Field observational data show that social pairs mated assortatively by head plumage, suggesting that these phenotypes are maintained by divergent mating preferences. In contrast, variation in all other traits and genetic markers could be explained by neutral diffusion, although weak ecological selection cannot be ruled out. Our results emphasize that assortative mating may maintain phenotypic differences independent of other processes shaping genome-wide variation, consistent with other recent findings that raise questions about the relative importance of mate choice, ecological selection and selectively neutral processes for divergent evolution.


Does magnetic resonance imaging compromise integrity of the blood-brain barrier?
It has been reported that a standard clinical procedure of magnetic resonance imaging (MRI) carried out experimentally in the rat caused temporary opening of the blood-brain barrier. Electron micrography indicated blood-to-brain movement of horseradish peroxidase, a protein tracer that does not normally permeate the barrier. In the present study, permeability-area products (PA), describing the limited permeation of blood-borne [14C]sucrose into brain parenchyma, were measured in anesthetized rats subjected either to 23 min of MRI, to osmotic barrier opening by intracarotid infusion of hypertonic arabinose, or to control conditions. Osmotic opening caused manyfold increases in PA whereas MRI produced no change. The proposal that MRI can compromise integrity of the blood-brain barrier is not supported by these findings.


Evolution of an Instrument: Measuring the Nursing Work Environment: A Scoping Review.
Background and Purpose: The Nursing Work Index (NWI) was developed in the 1980s to measure the nursing work environment (NWE). Instruments descended from the NWI continue to measure the NWE today. The purpose of this review was to identify instruments derived from the NWI, examine how they have been used and revised, and evaluate their ability to capture elements of the current work environment. Methods: A scoping literature review. Results: Forty articles were included. Instruments developed from the NWI have been translated into numerous languages and administered to hundreds of thousands of nurses globally. Conclusions: The study of the NWE remains extensive throughout the world. Future research should examine the factorial structure of instrument adaptions and ensure that items are relevant to contemporary nursing practice.


On the structure of a proposed mixed-valent analogue of the diiron subsite of [FeFe]-hydrogenase.
We show that a dinuclear assembly apparently providing the first example of a synthetic molecule exhibiting key features of the diiron subsite of [FeFe] hydrogenase, viz. CO-bridging of a coordinatively unsaturated, dithiolate-bridged mixed-valence diiron centre, is in fact a diamagnetic tetranuclear complex.


New insights into the chemistry of Coenzyme Q-0: A voltammetric and spectroscopic study.
Coenzyme Q-0 (CoQ-0) is the only Coenzyme Q lacking an isoprenoid group on the quinoid ring, a feature important for its physico-chemical properties. Here, the redox behavior of CoQ-0 in buffered and non-buffered aqueous media was examined. In buffered aqueous media CoQ-0 redox chemistry can be described by a 2-electron-2-proton redox scheme, characteristic for all benzoquinones. In non-buffered media the number of electrons involved in the electrode reaction of CoQ-0 is still 2; however, the number of protons involved varies between 0 and 2. This results in two additional voltammetric signals, attributed to 2-electrons-1H(+) and 2-electrons-0H(+) redox processes, in which mono- and di-anionic compounds of CoQ-0 are formed. In addition, CoQ-0 exhibits a complex chemistry in strong alkaline environment. The reaction of CoQ-0 and OH(-) anions generates several hydroxyl derivatives as products. Their structures were identified with HPLC/MS. The prevailing radical reaction mechanism was analyzed by electron paramagnetic resonance spectroscopy. The hydroxyl derivatives of CoQ-0 have a strong antioxidative potential and form stable complexes with Ca(2+) ions. In summary, our results allow mechanistic insights into the redox properties of CoQ-0 and its hydroxylated derivatives and provide hints on possible applications.


Isolation of a Bacillus subtilis mutant defective in constitutive O6-alkylguanine-DNA alkyltransferase.
A mutant of Bacillus subtilis defective in the constitutive activity of O6-alkylguanine-DNA alkyltransferase was isolated from a strain (ada-1) deficient in the adaptive response to DNA alkylation. Cells carrying the mutation dat-1 which was responsible for the defect in constitutive activity exhibited hypersensitivity for lethality and mutagenesis when challenged with methyl-nitroso compounds. The constitutive activity is independent of the adaptive response, and seems to function as a basal defense against environmental alkylating agents.


Ultra-processed food intake and incident venous thromboembolism risk: Prospective cohort study.
Ultra-processed food (UPF) intake has been associated with multiple health outcomes, but data on the association between UPF intake and venous thromboembolism (VTE) risk are lacking. We conducted this study to examine the association between UPF intake and the risk of incident VTE. This prospective cohort study was based on 186,323 participants free of baseline VTE from the UK Biobank. UPF intake was assessed by 24-h recall questionnaires. Data on incident VTE came from the nationwide inpatient and primary care datasets and the death registry. Cox proportional hazards regression was used to estimate the association between UPF intake and incident VTE risk. Multiplicative interactions and stratified analyses by age, sex, and body mass index were performed. During a 10.5-year (median) follow-up, 4235 incident VTE cases were diagnosed. After adjusting for covariates, the hazard ratio of VTE among individuals with the highest quintile of UPF intake was 1.05 (95% confidence interval [CI] 0.94, 1.17) for UPF in servings, 1.12 (95% CI 1.01, 1.24) in grams, 1.10 (95% CI 1.00, 1.22) in grams %, 1.21 (95% CI 1.10, 1.33) in energy, and 1.15 (95% CI 1.05, 1.27) in energy % compared to those in the lowest quintile. Age, sex, and body mass index did not modify the associations (Pinteraction > 0.05). Higher UPF intake was associated with a moderately increased risk of VTE.


Epidemiological Aspects of Oral Cancer in North Indian Population.
Head and neck cancers are common in the developing nations, especially in the south-east Asia. They account for the commonest malignancy type in India and pose a major public health concern. Oral cancer is the commonest of all head and neck malignancies. It is a prospective study done from January 2017 to July 2018 at a tertiary care centre with patient population ranging from 22 to 87 years of age. All the 128 patients, irrespective of their gender, with a proven malignancy of the oral cavity were observed for various epidemiological and disease characteristics. Oral cavity cancers were more common in males than females. It is prevalent in the age group of 60-70 years. The most common site of involvement is the buccal mucosa followed by the tongue. Tobacco chewing and smoking is the most significant risk factor associated. The most commonly involved neck nodes are level I and II lymph nodes. Oral cancers are common in Indian population. It presents at advanced stage due to poor socioeconomic status of the patients. However, it has also been seen in younger population owing to increasing use of tobacco products. Preventive strategy, based on the epidemiological characteristics, is the need of the hour to reduce the disease burden.


Economic evaluation of filgrastim, sargramostim, and sequential sargramostim and filgrastim after myelosuppressive chemotherapy.
Filgrastim alone and sequential sargramostim and filgrastim have been shown to be more effective than sargramostim alone in the mobilization of CD34(+) cells after myelosuppressive chemotherapy (MC). We sought to compare costs and resource use associated with these regimens. Data were collected prospectively alongside a multicenter, randomized trial of filgrastim, sargramostim, and sequential sargramostim and filgrastim. Direct medical costs were calculated for inpatient and outpatient visits and procedures, including administration of growth factors and MC. We followed 156 patients for 30 days or until initiation of high-dose chemotherapy. The main outcome measures were resource use and costs of inpatient and outpatient visits, platelet and red blood cell transfusions, antibiotic use, and apheresis procedures. Hospital admissions, red blood cell transfusions, and use of i.v. antibiotics were significantly more common in the sargramostim group than in the other treatment arms. In univariate and multivariable analyses, total costs were higher for patients receiving sargramostim alone than for patients in the other groups. Mean costs in multivariable analysis for the filgrastim and sequential sargramostim and filgrastim arms were not significantly different. Filgrastim alone and sequential sargramostim and filgrastim are less costly than sargramostim alone after MC, as well as therapeutically more beneficial.