Occurrence of resilin and its significance in the cuticle of Pennella elegans, a copepod parasite.
The cuticle of the region connecting the anterior and posterior half of the body of parasite shows two peculiarities. The epicuticle is folded and the lamellations in the outer region of the procuticle are wavy. Histochemical tests and investigations with fluorescent compounds showed that there is evidence of stabilization of the cuticle protein by formation of di- and trityrosine links as reported in the wing ligament cuticle of insects and elastic leg-hinge of the crayfish. By chromatography analysis, the above two amino acids were isolated and were found to be involved in the stabilization of resilin. In the light of above findings it is suggested that this may be related to this region serving as a flexible hinge.


Evaluating the potentials of randomised integrated control trial on tungiasis in a South-western Nigerian community.
Considering the fact that new, safe and incorporable treatment alternatives to therapeutic prophylaxis for tungiasis are lacking and sometimes proving difficult, this present study evaluated the potentials of integrating control approach involving focal premise treatment using 5% emulsifiable concentrate of cypermethrin, and topical application of Piper guineense oil with personal protection. Of the 90 houses selected, their floors were classified into paved rooms & unpaved verandas, paved verandas & unpaved rooms, paved rooms & verandas, and unpaved rooms & verandas, and tested for the presence of sand fleas using the sweeping and beating as well as soil collection and extraction by tullgren funnel method before fumigation. A total of 100 individuals partitioned into four groups of 25 individuals each were assigned 3%, 5%, 10% and 15% of P. guineense oil irrespective of stages of embedded fleas. The mean of sand fleas decreased from 1.14, 0.07, 0.21, and 1.66 to 0.37, 0.02, 0.09 and 1.08 after two weeks of single spray compared to the untreated location (p < 0.001). Furthermore, the mean lesions of stage I reduced to 0 after 2 to 6 days of 3%, 5%, 10% and 15% ointment application, and stage II and III after 10 to 12 days of 10% and 15% application respectively. The difference between tungiasis stages and exposure time for P. guineense ointment was highly significant p = 0.007 and p = 0.0002. Notable reduction in severity score of acute and chronic tungiasis was observed thus indicating effectiveness of the topical ointment. Hexanolic oil extracts of P. guineense in 10% and 15% concentration would kill embedded sand flea and ameliorate the sufferings in endemic settings.


Immune-spaying as an alternative to surgical spaying in Iberian×Duroc females: effect on carcass traits and meat quality characteristics.
The aim of this study was to assess the effect of immune-spaying on production traits and meat quality characteristics of fresh loin (longissimus dorsi) by comparing 3 groups of Iberian×Duroc females (N=12 per batch): surgically spayed, immune-spayed and entire females. Carcass traits and physicochemical parameters, including fatty acid profile, were investigated. The only carcass trait significantly affected by castration was the ham fat thickness, where both immune-spayed and surgically spayed females showed higher values against entire females (57±9.5mm, 62±2.5mm and 51±10.1mm, respectively; p<0.05). Furthermore, there were no significant differences in the quality parameters of fresh meat. These results are important regarding animal welfare, since in Europe, there is a plan to voluntarily end the surgical castration of pigs by 2018. Therefore, with an adequate vaccination protocol, immune-spaying might represent a good alternative to surgical spaying. The reliability of immune-spaying over long periods should be evaluated.


Physical activity, mood and the functioning of daily living A longitudinal study among former elite athletes and referents in middle and old age.
We studied whether factors related to type of sport participated in as young adults and level of and changes in physical activity later in life predict changes in mood as well as functioning during a 6-year follow-up. A cohort of male Finnish former athletes (N=504), referents (N=349) was followed up for changes in physical activity, in relation to subsequent self-reported mood and functioning of daily living in 1985, 1995, and 2001. The mean age of the cohort was 68.6 years in 2001. Multinomial logistic regression analysis was used to assess changes in mood and functioning between 1995 and 2001 in relation to baseline values and changes in exposure variables and covariates. A low level of physical activity in 1985 predicted a decrease in physical functioning between 1995 and 2001 in the lowest physical activity compared to the highest quintile as well as poor physical functioning at the end of follow-up in 2001. An increase in physical activity between 1985 and 1995 protected against onset of anxiety between 1995 and 2001. Physical activity for elderly seems to have an important role in reducing the progress of deficiencies in physical functioning and in preventing onset of anxiety.


Rational Construction of Heterostructured Cu3 P@TiO2 Nanoarray for High-Efficiency Electrochemical Nitrite Reduction to Ammonia.
Electroreduction of nitrite (NO2 - ) to valuable ammonia (NH3 ) offers a sustainable and green approach for NH3 synthesis. Here, a Cu3 P@TiO2 heterostructure is rationally constructed as an active catalyst for selective NO2 - -to-NH3 electroreduction, with rich nanosized Cu3 P anchored on a TiO2 nanoribbon array on Ti plate (Cu3 P@TiO2 /TP). When performed in the 0.1 m NaOH with 0.1 m NaNO2 , the Cu3 P@TiO2 /TP electrode obtains a large NH3 yield of 1583.4 µmol h-1 cm-2 and a high Faradaic efficiency of 97.1%. More importantly, Cu3 P@TiO2 /TP also delivers remarkable long-term stability for 50 h electrolysis. Theoretical calculations indicate that intermediate adsorption/conversion processes on Cu3 P@TiO2 interfaces are synergistically optimized, substantially facilitating the conversion of NO2 - -to-NH3 .


Improving risk assessment and prevention of pressure injuries during the implementation of a best practice clinical guideline.
To evaluate the indicators of quality of care in pressure injuries (PI) before and after 6 months of the implementation of the Good Clinical Practice Guideline of the Registered Nurses' Association of Ontario in the geriatric unit of the Hospital Universitari Santa Maria de Lleida. Longitudinal descriptive observational study throughout the implementation of the Good Clinical Practice Guideline (GCP) "Risk Assessment and Pressure Injury Prevention" carried out by the Ontario Nurses' Professional Association. The main variables - incidence and category of nosocomial pressure injuries, risk assessment of PI during the first 24hours of admission and risk level according to the EMINA scale, continuous assessment according to the risk of PI and special surface for pressure management recorded in the care plan - were extracted from the electronic medical records for subsequent descriptive analysis and hypothesis contrasting for comparison of proportions. A total of 154 subjects were included, most of them being women (57%), average age of 86 years and an average stay of admission of 8 days. With the implementation of the guide it was possible to improve, not always with statistical significance, the quality indicators: the incidence of PI decreased by 14.54%, risk assessments of PI 24hours after admission increased by 2.90%, while periodic risk assessments increased by 280.35%, recording 48.19% compared to 12.67% for the baseline situation. In addition, the recording of special surfaces in patients at risk of PI also increased by 13.33%. The implementation of the RNAO GCP improved the results related to the assessment and prevention of PI, with a positive impact on the quality of care indicators.


Effect of impeller type and agitation on the performance of pilot scale ASBR and AnSBBR applied to sanitary wastewater treatment.
The objective of this work was to assess the effect of agitation rate and impeller type in two mechanically stirred sequencing batch reactors: one containing granulated biomass (denominated ASBR) and the other immobilized biomass on polyurethane foam (denominated AnSBBR). Each configuration, with total volume of 1 m(3), treated 0.65 m(3) sanitary wastewater at ambient temperature in 8-h cycles. Three impeller types were assessed for each reactor configuration: flat-blade turbine impeller, 45 degrees -inclined-blade turbine impeller and helix impeller, as well as two agitation rates: 40 and 80 rpm, resulting in a combination of six experimental conditions. In addition, the ASBR was also operated at 20 rpm with a flat-blade turbine impeller and the AnSBBR was operated with a draft tube and helix impeller at 80 and 120 rpm. To quantify how impeller type and agitation rate relate to substrate consumption rate, results obtained during monitoring at the end of the cycle, as well as the time profiles during a cycle were analyzed. Increasing agitation rate from 40 rpm to 80 rpm in the AnSBBR improved substrate consumption rate whereas in the ASBR this increase destabilized the system, likely due to granule rupture caused by the higher agitation. The AnSBBR showed highest solids and substrate removal, highest kinetic constant and highest alkalinity production when using a helix impeller, 80 rpm, and no draft tube. The best condition for the ASBR was achieved with a flat-blade turbine impeller at 20 rpm. The presence of the draft tube in the AnSBBR did not show significant improvement in reactor efficiency. Furthermore, power consumption studies in these pilot scale reactors showed that power transfer required to improve mass transfer might be technically and economically feasible.


Purging of chronic myelogenous leukemia cells by retrovirally expressed anti-bcr-abl ribozymes with specific cellular compartmentalization.
In patients with chronic myelogenous leukemia (CML), abnormal expansion of myeloid cells is maintained by expression of the p210(bcr-abl) fusion protein. Thus, this protein and its mRNA represent primary targets to inhibit proliferation of these cells. Here we describe the properties of a ribozyme against the bcr-abl mRNA, expressed as a fusion transcript with the human U1 small nuclear RNA or the adenovirus VA1 RNA and delivered to the cells through retroviral vectors. These fusion ribozymes are specifically localized in the nucleus or in the cytoplasm, respectively. Transduction of 32D-LG7 myeloid cells, whose growth is IL-3 independent thanks to deregulated bcr-abl expression, imposed strong negative selective pressure on cell growth and induced restoration of an IL-3-dependent phenotype. Although expressed at a level similar to that of the U1-fusion ribozyme, the cytoplasmic VA1 ribozyme was a more powerful inhibitor of p210(bcr-abl) gene expression. In cells transduced with the vector expressing this ribozyme, the levels of the bcr-abl transcript were reduced up to 10(4)-fold, the p210(bcr-abl) protein became undetectable, and the cells underwent massive apoptosis when cultured in the absence of IL-3. Transduction of primary hematopoietic cells obtained from bone marrow of patients with CML resulted in remarkable reduction of bcr-abl mRNA levels, starting a few days after transduction. These results show the feasibility and efficacy of vector-expressed anti-bcr-abl ribozymes for purging of CML cells.


Effect of Cytomegalovirus Reactivation With or Without Acute Graft-Versus-Host Disease on the Risk of Nonrelapse Mortality.
Despite a strong association between acute graft-versus-host disease (GVHD) and cytomegalovirus reactivation (CMVR), the joint effect of acute GVHD and CMVR on nonrelapse mortality (NRM) has not been well studied. We evaluated the impact of CMVR on NRM stratified according to the development of acute GVHD using a landmark method. This study included 6078 patients who received their first allogeneic hematopoietic cell transplantation (HCT) with a preemptive strategy for CMVR between 2008 and 2017. The cumulative incidences of grade 2-4 acute GVHD (G24GVHD), CMVR by day 100, and CMV disease by day 365 were 37.3%, 52.1%, and 2.9%, respectively. Patients with G24GVHD were associated with the subsequent development of CMVR, and the presence of CMVR also increased the risk of G24GVHD. In a landmark analysis at day 65, the cumulative incidence of NRM at 1 year was 5.4%, 10.0%, 13.9%, and 19.7% in patients with G24GVHD-/CMVR-, G24GVHD-/CMVR+, G24GVHD+/CMVR-, and G24GVHD+/CMVR+, respectively. In a multivariate analysis, CMVR was respectively associated with an increased risk of NRM by day 365 in patients without G24GVHD (hazard ratio [HR], 1.59; 95% confidence interval [CI], 1.24-2.05; P < .001) and with G24GVHD (HR, 1.34; 95% CI, 1.06-1.70; P = .014), but the interaction between G24GVHD and CMVR was not significant (P = .326). Subgroup analyses suggested that the joint effect of acute GVHD and CMVR might vary according to the baseline characteristics. These data regarding the close relationship between acute GVHD and CMVR should provide important implications for the treatment strategy after HCT.


High quantum yield aminophenylboronic acid-functionalized N-doped carbon dots for highly selective hypochlorite ion detection.
High quantum yield 3-aminophenylboronic acid-functionalized nitrogen-doped carbon dots (GAAP-CDs) were fabricated using a simple hydrothermal route and used as a sensing probe for toxic hypochlorite (ClO-). The as-synthesized GAAP-CDs showed absorption peaks at 252, 297, and 370 nm and an emission peak at 375 nm with an excitation wavelength of 310 nm. The quantum yield of GAAP-CDs reached 58.28%, with no noticeable fluorescence change observed under high ionic strength conditions and a three-month long-term test. GAAP-CDs-based ClO- sensing was carried out by UV-vis absorbance and fluorescence spectroscopy; the detection limit was as low as 0.77 μM (linear range of 0-100 μM), and 0.50 μM (linear range of 0.1-100 μM), respectively. In addition, the as-synthesized GAAP-CDs showed excellent selectivity towards ClO- ions in the presence of various interfering chemicals. The satisfactory results from the proposed method of ClO- detection in tap water and drinking water samples, suggesting promising application of GAAP-CDs for ClO- detection.


Epinephrine's effect in varying concentrations on the end artery organ in an animal model.
To determine the concentration of epinephrine that causes end artery necrosis and ischemia. An experimental study. Animal Laboratory of Khordad Hospital, Tehran, Iran, from September 2007 to June 2008. This study was done on 54 ear flaps from 9 rabbits [9 (rabbits) x2 (ears) x3 (splits) =54] divided into three groups of 18 ear flaps each. Nine of the ear flaps were randomized as control and nine of them were randomized as study group. The different concentrations of epinephrine and lidocaine 1% were injected in study groups but only 1% lidocaine was injected in control groups. Study group 1 received 1/200000, group 2 received 1/100000 and group 3 received 1/500000 concentration of epinephrine respectively. There was no evidence of necrosis in the control and study groups 1 and 2. However, 4 out of 9 of the ear flaps showed clear evidence of tissue necrosis in group 3. Injecting epinephrine in 1/200000 and 1/100000 concentrations had no side effect for the end artery of the rabbit ear flaps, but 1/50000 concentration is shown that it can cause tissue necrosis. Further studies need to be done in order to find out the effects of epinephrine with the same concentrations on the human end artery.


Comparative Evaluation of Dentinal Microcrack Formation before and after Root Canal Preparation Using Rotary, Reciprocating, and Adaptive Instruments at Different Working Lengths-A Micro-computed Tomographic Study.
This study aimed to compare root dentinal microcrack formation after root canal shaping using rotary, reciprocating, and adaptive instruments at different working lengths using micro-computed tomographic imaging. One hundred eighty extracted mature mandibular molar mesial roots with 2 separate canals were selected. The mesial roots were resected at the cementoenamel junction and randomly divided into 4 groups (n = 45) based on the nickel-titanium file system used: ProTaper Universal (Dentsply Maillefer, Ballaigues, Switzerland), ProTaper Gold (Dentsply Maillefer), Twisted File Adaptive (SybronEndo, Orange, CA), and Reciproc Blue (VDW, Munich, Germany). Each of the 4 groups were then subdivided into 3 groups (n = 15) depending on the working length used for root canal preparation (ie, instrumentation 1 mm short, flush, and 1 mm beyond the major apical foramen). The roots were imaged with micro-computed tomographic scanning before and after root canal preparation. The cross-sectional images generated were screened to detect the presence of new microcracks. The ProTaper Universal system significantly increased the number of postinstrumentation microcracks at all working lengths (P ≤ .05). No significant increase (P > .05) in postinstrumentation microcracks was observed in the ProTaper Gold, Twisted File Adaptive, or Reciproc Blue groups. Rotary instrumentation induced a higher number of dentinal microcracks compared with reciprocating and adaptive instruments. Instrumentation at different working lengths did not significantly influence the formation of dentinal microcracks.


Fibrosing interstitial lung disease in primary Sjogren syndrome.
Interstitial lung disease (ILD) represents the main pulmonary involvement in primary Sjogren syndrome (pSS). A proportion of patients with pSS develop a progressive fibrosing form of ILD, but no data are available about the prevalence of these patterns in pSS patients. Aim of this monocentric, cross-sectional study was to investigate the prevalence of fibrosing patterns in pSS patients with ILD. All consecutive patients fulfilling classification criteria for pSS with a new or previous diagnosis of ILD were enrolled in the study. Diagnosis of ILD was always performed by mean of HRCT and specific patterns were identified according to current classification criteria and divided in two groups according to the detection of a fibrotic pattern. Thirty-four pSS-ILD patients were enrolled in the study (males/females 3/31, median age 69.5 years, median pSS duration 47.5 months). Fibrotic pattern was detected in 52.9% of patients, namely: UIP (13 patients, 38.2%), fibrotic NSIP (4, 11.8%), fibrotic OP (1 2.9%) and group 2 (16 pts, 47.1%) including NSIP (6, 17.6%), OP (4, 11.8%), LIP (2, 5.9%) and unclassifiable (4, 11.8%). These patients were younger and with a shorter pSS duration at ILD diagnosis, in particular ILD diagnosis was prior or concurrent to pSS in 83.3% of cases compared to 62.5% in the group of nonfibrotic pattern (P<0.05). Our data suggest a high prevalence of this pulmonary clinical phenotype in pSS-ILD patients. Since the course of progressive fibrosing pneumonia generally results in respiratory failure, this result could be worthy of further studies.


Morphometry and microanatomy of the barbels of the common sawshark Pristiophorus cirratus (Pristiophoridae): implications for pristiophorid behaviour.
The internal anatomy of the barbels of the common sawshark Pristiophorus cirratus was examined with light microscopy to clarify their sensory role. No sensory structures such as taste buds (chemoreception), ampullae of Lorenzini (electroreception) or free neuromasts (lateral line mechanoreception) could be located in the barbels. The presence of bundles of nerve fibres, however, indicates a tactile function for the barbels. Conveyance of information regarding potentially damaging stimuli (nociception) and temperature (thermoception) cannot be excluded at this stage. It is hypothesized that the barbels are used by P. cirratus to locate prey in both the water column and on the substratum via wake detection and sensing changes in surface texture. The barbels may also be involved in the detection of water currents for rheotaxis. Regression analyses on P. cirratus morphometric data showed that the width of the rostrum at two sections (the barbels and the rostrum tip) does not significantly correlate with total length. The regression analyses also suggested that the barbels of P. cirratus may be lateralised.


Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina.
Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel in vivo gene silencing approach in the rat retina. siRNA designed for knockdown of the house keeping gene Gapdh or four retinal cell type-specific genes (Nefl, Pvalb, Rho and Opn1sw) was injected into the vitreous body, and their retinal mRNA levels were quantified using real-time PCR. Intravitreal injection of siRNA for Gapdh resulted in approximately 40-70% reduction in its retinal mRNA levels, which lasted throughout a 9-day study period. Furthermore, all the selected retinal specific genes were efficiently down-regulated by 60-90% following intravitreal injection, suggesting injected siRNA penetrated into major retinal cell types. These findings were consistent with uniform distribution of a fluorescence-labeled siRNA injected into the vitreous body. Interestingly, gene silencing of Grin1, a core subunit of NMDA receptor, was accompanied by significant prevention from NMDA-induced retinal ganglion cell death. Thus, we provide single intravitreal injection of Accell siRNA as a versatile technique for robust and sustainable in vivo retinal gene silencing to characterize their biological functions under physiological and pathophysiological conditions.


Portable all-in-one automated microfluidic system (PAMICON) with 3D-printed chip using novel fluid control mechanism.
State-of-the-art microfluidic systems rely on relatively expensive and bulky off-chip infrastructures. The core of a system-the microfluidic chip-requires a clean room and dedicated skills to be fabricated. Thus, state-of-the-art microfluidic systems are barely accessible, especially for the do-it-yourself (DIY) community or enthusiasts. Recent emerging technology-3D-printing-has shown promise to fabricate microfluidic chips more simply, but the resulting chip is mainly hardened and single-layered and can hardly replace the state-of-the-art Polydimethylsiloxane (PDMS) chip. There exists no convenient fluidic control mechanism yet suitable for the hardened single-layered chip, and particularly, the hardened single-layered chip cannot replicate the pneumatic valve-an essential actuator for automatically controlled microfluidics. Instead, 3D-printable non-pneumatic or manually actuated valve designs are reported, but their application is limited. Here, we present a low-cost accessible all-in-one portable microfluidic system, which uses an easy-to-print single-layered 3D-printed microfluidic chip along with a novel active control mechanism for fluids to enable more applications. This active control mechanism is based on air or gas interception and can, e.g., block, direct, and transport fluid. As a demonstration, we show the system can automatically control the fluid in microfluidic chips, which we designed and printed with a consumer-grade 3D-printer. The system is comparably compact and can automatically perform user-programmed experiments. All operations can be done directly on the system with no additional host device required. This work could support the spread of low budget accessible microfluidic systems as portable, usable on-the-go devices and increase the application field of 3D-printed microfluidic devices.


Insulin controls food intake and energy balance via NPY neurons.
Insulin signaling in the brain has been implicated in the control of satiety, glucose homeostasis and energy balance. However, insulin signaling is dispensable in energy homeostasis controlling AgRP or POMC neurons and it is unclear which other neurons regulate these effects. Here we describe an ancient insulin/NPY neuronal network that governs energy homeostasis across phyla. To address the role of insulin action specifically in NPY neurons, we generated a variety of models by selectively removing insulin signaling in NPY neurons in flies and mice and testing the consequences on energy homeostasis. By specifically targeting the insulin receptor in both fly and mouse NPY expressing neurons, we found NPY-specific insulin signaling controls food intake and energy expenditure, and lack of insulin signaling in NPY neurons leads to increased energy stores and an obese phenotype. Additionally, the lack of insulin signaling in NPY neurons leads to a dysregulation of GH/IGF-1 axis and to altered insulin sensitivity. Taken together, these results suggest that insulin actions in NPY neurons is critical for maintaining energy balance and an impairment of this pathway may be causally linked to the development of metabolic diseases.


Label propagation via local geometry preserving for deep semi-supervised image recognition.
In this paper, we propose a novel transductive pseudo-labeling based method for deep semi-supervised image recognition. Inspired from the superiority of pseudo labels inferred by label propagation compared with those inferred from network, we argue that information flow from labeled data to unlabeled data should be kept noiseless and with minimum loss. Previous research works use scarce labeled data for feature learning and solely consider the relationship between two feature vectors to construct the similarity graph in feature space, which causes two problems that ultimately lead to noisy and incomplete information flow from labeled data to unlabeled data. The first problem is that the learned feature mapping is highly likely to be biased and can easily over-fit noise. The second problem is the loss of local geometry information in feature space during label propagation. Accordingly, we firstly propose to incorporate self-supervised learning into feature learning for cleaner information flow in feature space during subsequent label propagation. Secondly, we propose to use reconstruction concept to measure pairwise similarity in feature space, such that local geometry information can be preserved. Ablation study confirms synergistic effects from features learned with self-supervision and similarity graph with local geometry preserving. Extensive experiments conducted on benchmark datasets have verified the effectiveness of our proposed method.


Ovarian malignant melanoma: unusual presentation of a solitary metastasis.
Malignant melanomas of the ovary are rare with unpredictable clinical and biological behavior. We present this case report of a metastatic ovarian malignant melanoma, where the ovary was the first site of relapse after 10 years of remission. A 58-year-old postmenopausal woman was incidentally found to have an abdominal mass, 10 years after removal of a cutaneous malignant melanoma lesion. Ultrasound and CT scan revealed an adnexal mass, which was found to be a solitary ovarian tumor at laparotomy. Histology confirmed malignant melanoma metastasis. Relapse after a prolonged period of remission, presenting, as an isolated ovarian metastasis is an unusual presentation of malignant melanoma. As illustrated by this case report, a differential diagnosis of a metastatic malignant melanoma must be considered.


The plastoglobule-localized protein AtABC1K6 is a Mn2+-dependent kinase necessary for timely transition to reproductive growth.
The Absence of bc1 Complex (ABC1) is an ancient, atypical protein kinase family that emerged prior to the archaeal-eubacterial divergence. Loss-of-function mutants in ABC1 genes are linked to respiratory defects in microbes and humans and to compromised photosynthetic performance and stress tolerance in plants. However, demonstration of protein kinase activity remains elusive, hampering their study. Here, we investigate a homolog from Arabidopsis thaliana, AtABC1K6, and demonstrate in vitro autophosphorylation activity, which we replicate with a human ABC1 ortholog. We also show that AtABC1K6 protein kinase activity requires an atypical buffer composition, including Mn2+ as a divalent cation cofactor and a low salt concentration. AtABC1K6 associates with plastoglobule lipid droplets of A. thaliana chloroplasts, along with five paralogs. We show that the protein kinase activity associated with isolated A. thaliana plastoglobules was inhibited at higher salt concentrations, but could accommodate Mg2+ as well as Mn2+, indicating salt sensitivity, but not the requirement for Mn2+, may be a general characteristic of ABC1 proteins. Finally, loss of functional AtABC1K6 impairs the developmental transition from vegetative to reproductive growth. This phenotype was complemented by the wild-type sequence of AtABC1K6, but not by a kinase-dead point mutant in the unique Ala-triad of the ATP-binding pocket, demonstrating the physiological relevance of the protein's kinase activity. We suggest that ABC1s are bona fide protein kinases with a unique regulatory mechanism. Our results open the door to detailed functional and mechanistic studies of ABC1 proteins and plastoglobules.


Caudatin induces cell cycle arrest and caspase-dependent apoptosis in HepG2 cell.
In the present study, we investigate the anti-cancer activity and mechanism of caudatin, the C-21 steroidal glycosides, on human hepatoma cell line HepG2. The MTT assay and flow cytometry were used to evaluate HepG2 cell proliferation and cell cycle. Annexin-V/PI and DAPI staining were used to investigate cell apoptosis. Western blotting analysis was used to evaluate the expression levels of proteins. It is found that caudatin inhibits HepG2 cell growth and induces of G0/G1 phase arrest in a dose dependent manner, which is associated with a decreased in the expression of cyclinD1 and increased the levels of p21 and p53. HepG2 cells dealing with caudatin showed typical characteristics of apoptosis. Western blotting analysis indicated that the levels of Bcl-2 were down-regulated after caudatin treatment, whereas the expression of Bax was up-regulated. Furthermore, caudatin-induced apoptosis was accompanied by activation of caspase-3, -9, and poly(ADP-Ribose) Polymerase (PARP). Treatment with caudatin also induced phosphorylation of extracellular-signal regulating kinase (ERK) and c-Jun N-terminal kinase (JNK). These results demonstrate that caudatin inhibits cell proliferation via DNA synthesis reduction and induces caspase-dependent apoptosis in HepG2 cell. Activation of ERK and JNK may be involved in caudatin-induced hepatoma cell apoptosis.


Evaluation of fatty acid profile, color characteristics, oxidative quality and stability of common Kilka (Clupeonella cultriventris caspia) oil obtained by various extraction techniques.
This study aimed to evaluate the impact of various oil extraction procedures on yield, color properties, fatty acid profile and oxidative stability of common Kilka (Clupeonella cultriventris caspia) oil. The supercritical fluid extraction (SFE) with carbon dioxide, than other techniques (wet reduction, ammonia and enzymatic extractions), showed the highest oil yield (96.94%), the best oxidative and color characteristics, with 0.68 mg KOH/g oil, 2.62 meq O2/kg oil, 6.5 and 5.36 in terms of acid value (AV), peroxide value (PV), yellowness and redness, respectively, and have the highest total of unsaturated fatty acids average (10.33), especially the omega-3 family. Furthermore, the oil obtained by SFE had the best oxidative parameters; AV, PV and thiobarbituric acid values, during the whole 90 days of storage time at 4 °C. Therefore, the SFE is the most effective procedure at obtaining the oil from common Kilka tissue than other methods studied in this research.


Validation of the North American Chest Pain Rule in Prediction of Very Low-Risk Chest Pain; a Diagnostic Accuracy Study.
Acute coronary syndrome accounts for more than 15% of the chest pains. Recently, Hess et al. developed North American Chest Pain Rule (NACPR) to identify very low-risk patients who can be safely discharged from emergency department (ED). The present study aimed to validate this rule in EDs of two academic hospitals. A prospective diagnostic accuracy study was conducted on consecutive patients 24 years of age and older presenting to the ED with the chief complaint of acute chest pain, during March 2013 to June 2013. Chest pain characteristics, cardiac history, electrocardiogram findings, and cardiac biomarker measurement of patients were collected and screening performance characteristics of NACPR with 95% confidence interval were calculated using SPSS 21. From 400 eligible patients with completed follow up, 69 (17.25 %) developed myocardial infarction, 121 (30.25%) underwent coronary revascularization, and 4 (2%) died because of cardiac or unidentifiable causes. By using NACPR, 34 (8.50%) of all the patients could be considered very low- risk and discharged after a brief ED assessment. Among these patients, none developed above-mentioned adverse outcomes within 30 days. Sensitivity, specificity, positive prediction value, and negative prediction value of the rule were 100% (95% CI: 87.35 - 100.00), 45.35 (95% CI: 40.19 - 50.61), 14.52 (95% CI: 10.40 - 19.85), and 100 (95% CI: 97.18 - 100.00), respectively. The present multicenter study showed that NACPR is a good screening tool for early discharge of patients with very low-risk chest pain from ED.


Tuning local chemistry of P2 layered-oxide cathode for high energy and long cycles of sodium-ion battery.
Layered transition-metal oxides have attracted intensive interest for cathode materials of sodium-ion batteries. However, they are hindered by the limited capacity and inferior phase transition due to the gliding of transition-metal layers upon Na+ extraction and insertion in the cathode materials. Here, we report that the large-sized K+ is riveted in the prismatic Na+ sites of P2-Na0.612K0.056MnO2 to enable more thermodynamically favorable Na+ vacancies. The Mn-O bonds are reinforced to reduce phase transition during charge and discharge. 0.901 Na+ per formula are reversibly extracted and inserted, in which only the two-phase transition of P2 ↔ P'2 occurs at low voltages. It exhibits the highest specific capacity of 240.5 mAh g-1 and energy density of 654 Wh kg-1 based on the redox of Mn3+/Mn4+, and a capacity retention of 98.2% after 100 cycles. This investigation will shed lights on the tuneable chemical environments of transition-metal oxides for advanced cathode materials and promote the development of sodium-ion batteries.


Neuropathological Characterization of Dolphin Morbillivirus Infection in Cetaceans Stranded in Italy.
Cetacean morbillivirus (CeMV) is responsible for epidemic and endemic fatalities in free-ranging cetaceans. Neuro-inflammation sustained by CeMV is a leading cause of death in stranded cetaceans. A novel dolphin morbillivirus (DMV) strain of Atlantic origin circulating in Italian waters since early 2016 has caused acute/subacute lesions associated with positive immunolabelling of the virus. To date, myelin damage has not been fully documented and investigated in cetaceans. This study describes neuropathological findings in the brain tissue of 31 cetaceans found stranded along the Italian coastline and positive for DMV infection on molecular testing. Cell changes in the areas of myelinopathy were revealed by double indirect immunofluorescence. The most frequent DMV-associated lesions were astro-microgliosis, neuronal necrosis, spongiosis, malacia, and non-suppurative meningoencephalitis. Myelin reduction and areas of demyelination were revealed by means of a specific myelin biomarker. Morbilliviral antigen immunolabelling was mainly observed in neurons and microglial cells, in association with a marked activation of microglia and astrocytes. These findings extend our knowledge of DMV-associated brain lesions and shed light on their pathogenesis.


Fluid and food intake during professional men's and women's road-cycling tours.
To quantify the fluid and food consumed during a men's and women's professional road-cycling tour. Eight men (age 25 +/- 5 y, body mass 71.4 +/- 7.4 kg, and height 177.4 +/- 4.5 cm) and 6 women (age 26 +/- 4 y, body mass 62.5 +/- 5.6 kg, and height 170.4 +/- 5.2 cm) of the Australian Institute of Sport Road Cycling squads participated in the study. The men competed in the 6-d Tour Down Under (Adelaide, Australia), and the women, in the 10-d Tour De L'Aude (Aude, France). Body mass was recorded before and immediately after the race. Cyclists recalled the number of water bottles and amount of food they had consumed. Men and women recorded body-mass losses of approximately 2 kg (2.8% body mass) and 1.5 kg (2.6% body mass), respectively, per stage during the long road races. Men had an average fluid intake of 1.0 L/h, whereas women only consumed on average 0.4 L/h. In addition, men consumed CHO at the rate suggested by dietitians (average CHO intake of 48 g/h), but again the women failed to reach recommendations, with an average intake of approximately 21 g/h during a road stage. Men appeared to drink and eat during racing in accordance with current nutritional recommendations, but women failed to reach these guidelines. Both men and women finished their races with a body-mass loss of approximately 2.6% to 2.8%. Further research is required to determine the impact of this loss on road-cycling performance and thermoregulation.


Modelling the response of auditory midbrain neurons in the grassfrog to temporally structured monaural stimuli.
In a previous paper [Van Stokkum and Gielen, Hear. Res. 41, 71-86, 1989] a model was presented to describe the processing of monaural stimuli by the auditory periphery of the grassfrog. The main components of this model were: a middle ear filter, transduction and tuning of the haircell, short-term adaptation, action potential (event) generation with refractory properties, and spatiotemporal integration of converging inputs. The model is now extended to model auditory midbrain neurons as third order neurons. The mechanisms that generate selectivity for temporal characteristics of sound are adaptation, coincidence detection of second order neurons, temporal integration of third order neurons, and most important, event generation of the first, second and third order model neurons. Variation of the parameters of the model successfully reproduces the range of response patterns which have been obtained from eighth nerve fibres, dorsal medullary nucleus neurons, and torus semicircularis neurons without inhibition. With a single set of parameters the output of the model in response to a set of spectrally and temporally structured stimuli qualitatively resembles the responses of a single neuron to all these stimuli. In this way the responses to the different stimuli are synthesized into a framework, which functionally describes the neuron.


Bezlotoxumab Therapy for Recurrent Clostridium difficile Infection in an Ulcerative Colitis Patient.
Clostridium difficile infection (CDI) is the most common infectious cause of nosocomial diarrhea, comprising 10%-20% of all cases. CDI is a significant complication in patients with inflammatory bowel disease (IBD). New monoclonal antibody therapies have emerged as leading treatment options for recurrent CDI (rCDI). Bezlotoxumab, a novel monoclonal antibody, has shown success in decreasing the recurrence rates of patients with rCDI. However, data extrapolating diminished rCDI in patients with concomitant IBD is limited. A single infusion of bezlotoxumab @ 10mg/kg was given with fidaxomicin 200mg for 10 days in a patient with rCDI and ulcerative colitis. The patient's symptoms improved, inflammatory markers normalized, and she has remained asymptomatic for twelve months. This case supports the findings in the MODIFY I/II trials that Bezlotoxumab is a viable treatment option of rCDI in IBD patients.


Use of intravenous FK506 to treat acute rejection in simultaneous pancreas-kidney transplant recipients on maintenance oral FK506.
Most recipients of simultaneous pancreas-kidney transplants experience acute rejection in the early postoperative course. We report our experience with four recipients of simultaneous pancreas-kidney transplants with acute rejection who were effectively treated with the combination of intravenous and oral FK506 therapy. This spared these patients an extra course of monoclonal or polyclonal antibody.


Accidental poisoning with podophyllin.
Podophyllin is available without prescription and is commonly used in the topical treatment of warts. We describe a case of accidental ingestion of podophyllin, mistaken for cough linctus, and its resultant toxicity.


Direct Zinc Determination in Brazilian Sugar Cane Spirit by Solid-Phase Extraction Using Moringa oleifera Husks in a Flow System with Detection by FAAS.
This paper reports a method for the determination of zinc in Brazilian sugar cane spirit, (cachaça in Portuguese), using solid-phase extraction with a flow injection analysis system and detection by FAAS. The sorbent material used was activated carbon obtained from Moringa oleifera husks. Flow and chemical variables of the proposed system were optimized through multivariate designs. The factors selected were sorbent mass, sample pH, sample flow rate, and eluent concentration. The optimum extraction conditions were obtained using a sample pH of 4.0, a sample flow rate of 6.0 mL min(-1), 30.0 mg of sorbent mass, and 1.0 mol L(-1) HNO(3) as the eluent at a flow rate of 4.0 mL min(-1). The limit of detection for zinc was 1.9 μg L(-1), and the precision was below 0.82% (20.0 μg L(-1), n = 7). The analytical curve was linear from 2 to 50 μg L(-1), with a correlation coefficient of 0.9996. The method developed was successfully applied to spiked Brazilian sugar cane spirit, and accuracy was assessed through recovery tests, with results ranging from 83% to 100%.


Human Epidermal Growth Factor Receptor-2 Gastric Adenocarcinoma: Expanding Therapy of a Recognized Target.
Human epidermal growth factor receptor-2 (HER2) is a well-known cancer target. Many HER2-targeted agents are marketed and being investigated. Unfortunately, these therapies lack consistent responses and outcomes amongst different tumors. Questions remain as to why HER2 biology is different in different tumor types. Gastric adenocarcinomas (GACs) demonstrate both intra- and inter-tumor HER2 expression heterogeneity and show discordance amongst primary and metastatic disease sites. This creates barriers in determining HER2 agents' effectiveness and contributes to the failure of some HER2-targeted agents in the treatment of HER2-positive advanced GACs. Trastuzumab deruxtecan, an antibody drug conjugate of trastuzumab with a topoisomerase inhibitor, was recently approved for the treatment of refractory HER2-positive advanced GAC patients. There are exciting and newer therapies under investigation. Examining resistance patterns (both adaptive and acquired) along with establishing a better understanding of the intra- and inter-tumor heterogeneity is necessary to ensure successful progress. Here we review the current status of HER2-targeted therapy in GACs. We additionally review newer therapies under investigation and their potential role in HER2 GACs.


A Review and Summary of Patients with Symptomatic Postoperative Discal Pseudocysts of the Lumbar Spine.
Postoperative discal pseudocyst (PDP) is a rare complication after discectomy. This study aimed to summarize the characteristics, pathological mechanisms and management of PDPs. Nine patients with PDP who received surgical treatment at our institution from January 2014 to December 2021 were retrospectively reviewed. A systematic review of the literature on PDP was performed. The demographic data, clinical and imaging features, surgical options and patient prognosis were analyzed. Among the nine patients treated at our center, seven were male and two were female. The mean patient age (± standard deviation) at the time of surgery was 28.3 ± 5.7 years (range 18-37 years). The first operation performed on seven patients was percutaneous endoscopic transforaminal discectomy (PETD) and two patients underwent microdiscectomy. The time to conservative treatment before surgical intervention was 20 ± 9.2 days. In three cases, the disc cysts were located in L4/5 and in six cases the lesions were located in L5/S1. Intervertebral disc cyst interventions included foraminal scope (three cases), open discectomy (three cases), conservative treatment with a quadrant channel (one case) and CT-guided puncture (one case). All patients fully recovered after surgery and the mean follow-up time was 3.5 ± 2.1 years. A literature review identified 14 relevant articles that reported 43 PDP cases of PDP. PDP occurs in Asian males with mild intervertebral disc degeneration and occurs 1 month after discectomy. Treatment should be based on specific patient scenarios. Conservative treatment is necessary and surgery should be performed with caution.


High Serum Alkaline Phosphatase Flare after First-Line Androgen Deprivation Therapy Predicts Poor Prognosis in Metastatic Prostate Cancer Patients Treated with Second-Generation Androgen Receptor Targeted Therapy.
To determine whether an alkaline phosphatase (ALP) flare after androgen deprivation therapy (ADT) is associated with the treatment response in castration-resistant prostate cancer (CRPC) and predicts the prognosis of metastatic prostate cancer (PCa) patients. One hundred and nineteen patients diagnosed with metastatic PCa between 2008 and 2017 were retrospectively studied. The ALP flare ratio was calculated as the ratio of ALP levels 1 month after beginning ADT to ALP levels at diagnosis. The association of the ALP flare ratio with the prostate-specific antigen (PSA) response to CRPC treatment (second-generation androgen receptor targeted therapy (ART) or docetaxel), time to CRPC, and overall survival (OS) were investigated. The time to CRPC and OS was significantly longer in patients with an ALP flare ratio less than 1.33 compared to a ratio more than 1.33. No difference in PSA response was seen regarding the ALP flare ratio in both ART and docetaxel treatment. Second-generation ART-treated patients with a low ALP flare ratio showed longer OS than those with a higher ALP flare ratio (p=0.0367). However, no difference was seen between a high and low ALP flare ratio (p=0.8054) in docetaxel-treated patients. The ALP flare ratio was the most significant prognostic factor for OS (p < 0.0001). A higher ALP flare ratio after first-line ADT was a significant prognostic factor in metastatic PCa, especially in patients treated with second-generation ART for CRPC. Chemotherapy for patients with a higher ALP flare ratio 1 month after induction of ADT may be a clinically relevant decision.


Patterns of blood mercury variation in two long-distance migratory thrushes on Mount Mansfield, Vermont.
We investigated mercury (Hg) blood concentrations in Bicknell's thrush (Catharus bicknelli) and Swainson's thrush (C. ustulatus), congeneric long-distance migratory songbirds, from 2000-2017 at a montane forest site in north-central Vermont. We analyzed variation in blood Hg of both species using mixed-effects models, incorporating atmospheric wet Hg deposition data from a nearby sampling location. Although Hg deposition varied among years and seasonally, we detected no temporal trend in either atmospheric deposition or blood Hg, nor evidence of a relationship between the two. Sampling date had the strongest effect on blood Hg concentration, which declined seasonally, followed by age and sex of the individual. The data did not support an effect of species. We believe that the absence of a clear relationship between local atmospheric deposition and thrush blood Hg concentrations suggests that Hg cycling dynamics, mechanisms of transfer, and timing of uptake by montane forest biota are complex and poorly understood. The blood Hg concentrations of ~0.07-0.1 μg/g we documented in Bicknell's and Swainson's thrush are below those found to negatively impact physiological or reproductive endpoints in other invertivorous terrestrial passerines. To better evaluate the validity of Bicknell's thrush as a bioindicator of MeHg availability in montane forest ecosystems, we recommend (1) effects-based investigations, (2) a more robust understanding of Hg and MeHg cycling, (3) more clear geospatial and temporal links between Hg deposition and biotic uptake, and (4) more thorough documentation of Hg burdens across the species' annual cycle.


TLR2-dependent modulation of dendritic cells by LT-IIa-B5, a novel mucosal adjuvant derived from a type II heat-labile enterotoxin.
A host of human pathogens invades the body at mucosal surfaces. Yet, strong, protective mucosal immune responses directed against those pathogens routinely cannot be induced without the use of adjuvants. Although the strongest mucosal adjuvants are members of the family of HLTs, the inherent toxicities of HLT holotoxins preclude their clinical use. Herein, it is shown that LT-IIa-B(5) enhances mucosal immune responses by modulating activities of DCs. i.n. immunization of mice with OVA in the presence of LT-IIa-B(5) recruited DCs to the NALT and significantly increased uptake of OVA by those DCs. Furthermore, LT-IIa-B(5) increased expression of CCR7 by DCs, which mediated enhanced migration of the cells from the NALT to the draining CLNs. LT-IIa-B(5) also enhanced maturation of DCs, as revealed by increased surface expression of CD40, CD80, and CD86. Ag-specific CD4(+) T cell proliferation was augmented in the CLNs of mice that had received i.n. LT-IIa-B(5). Finally, when used as an i.n. adjuvant, LT-IIa-B(5) dramatically increased the levels of OVA-specific salivary IgA and OVA-specific serum IgG. Strikingly, each of the activities induced by LT-IIa-B(5) was strictly TLR2-dependent. The data strongly suggest that the immunomodulatory properties of LT-IIa-B(5) depend on the productive modulation of mucosal DCs. Notably, this is the first report for any HLT to demonstrate in vivo the elicitation of strong, TLR2-dependent modulatory effects on DCs with respect to adjuvanticity.


A Randomized Phase 2 Study of Gefitinib With or Without Pemetrexed as First-line Treatment in Nonsquamous NSCLC With EGFR Mutation: Final Overall Survival and Biomarker Analysis.
Clinical studies have shown that a combination of a tyrosine kinase inhibitor (TKI) and pemetrexed overcame acquired resistance to epidermal growth factor receptor (EGFR) TKI in NSCLC. Previously, pemetrexed+gefintib (P+G) had improved progression-free survival (PFS) compared with gefitinib. We present OS, updated PFS, biomarker analysis, and safety of P+G versus gefitinib. This was a phase 2, multicenter, randomized study conducted in East Asian patients with advanced nonsquamous NSCLC with EGFR mutations. Patients were randomized (2:1) to receive P+G (500 mg/m2 intravenously 3-weekly + 250 mg/day orally) or gefitinib. In total, 191 patients (P+G, n=126; gefitinib, n=65) comprised the intent-to-treat and safety populations. Median OS was 43.4 months in P+G versus 36.8 months in gefitinib arm; adjusted HR 0.77 (95% CI, 0.5-1.2); one-sided P=0.105. Median PFS was significantly longer in the P+G (16.2 months) versus gefitinib arm (11.1 months); adjusted HR 0.67 (95% CI, 0.5-0.9); one-sided P=0.009. In the P+G and gefitinib arms, median PFS was 22.6 and 11.0 months, respectively, in patients with low thymidylate synthase (TS) expression, and 12.6 and 9.9 months, respectively, in patients with high TS expression. Common second-line post-discontinuation systemic therapies were EGFR-TKIs and chemotherapy. Most patients experienced at least one adverse event. Addition of pemetrexed to EGFR TKI gefitinib resulted in significantly improved PFS and numerically longer OS compared with gefitinib in treatment-naïve patients with EGFR-mutated advanced nonsquamous NSCLC. Low TS expression appeared to be a good predictor for treatment outcomes.


2-Chloro-4,6-bis-(piperidin-1-yl)-1,3,5-triazine.
The title compound, C(13)H(20)ClN(5), crystallizes with two mol-ecules in the asymmetric unit. The piperidine rings in both mol-ecules adopt chair conformations. Weak π-π inter-actions [centroid-centroid distance = 3.9815 (8) Å] are observed in the crystal structure.


Differential Phospho-Signatures in Blood Cells Identify LRRK2 G2019S Carriers in Parkinson's Disease.
The clinicopathological phenotype of G2019S LRRK2-associated Parkinson's disease (L2PD) is similar to idiopathic Parkinson's disease (iPD), and G2019S LRRK2 nonmanifesting carriers (L2NMCs) are at increased risk for development of PD. With various therapeutic strategies in the clinical and preclinical pipeline, there is an urgent need to identify biomarkers that can aid early diagnosis and patient enrichment for ongoing and future LRRK2-targeted trials. The objective of this work was to investigate differential protein and phospho-protein changes related to G2019S mutant LRRK2 in peripheral blood mononuclear cells from G2019S L2PD patients and G2019S L2NMCs, identify specific phospho-protein changes associated with the G2019S mutation and with disease status, and compare findings with patients with iPD. We performed an unbiased phospho-proteomic study by isobaric label-based mass spectrometry using peripheral blood mononuclear cell group pools from a LRRK2 cohort from Spain encompassing patients with G2019S L2PD (n = 20), G2019S L2NMCs (n = 20), healthy control subjects (n = 30), patients with iPD (n = 15), patients with R1441G L2PD (n = 5), and R1441G L2NMCs (n = 3) (total N = 93). Comparing G2019S carriers with healthy controls, we identified phospho-protein changes associated with the G2019S mutation. Moreover, we uncovered a specific G2019S phospho-signature that changes with disease status and can discriminate patients with G2019S L2PD, G2019S L2NMCs, and healthy controls. Although patients with iPD showed a differential phospho-proteomic profile, biological enrichment analyses revealed similar changes in deregulated pathways across the three groups. We found a differential phospho-signature associated with LRRK2 G2019S for which, consistent with disease status, the phospho-profile from PD at-risk G2019S L2NMCs was more similar to healthy controls than patients with G2019S L2PD with the manifested disease. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.


Construction of Organelle-Like Architecture by Dynamic DNA Assembly in Living Cells.
The design of controllable dynamic systems is vital for the construction of organelle-like architectures in living cells, but has proven difficult due to the lack of control over defined topological transformation of self-assembled structures. Herein, we report a DNA based dynamic assembly system that achieves lysosomal acidic microenvironment specifically inducing topological transformation from nanoparticles to organelle-like hydrogel architecture in living cells. Designer DNA nanoparticles are constructed from double-stranded DNA with cytosine-rich stick ends (C-monomer) and are internalized into cells through lysosomal pathway. The lysosomal acidic microenvironment can activate the assembly of DNA monomers, inducing transformation from nanoparticles to micro-sized organelle-like hydrogel which could further escape into cytoplasm. We show how the hydrogel regulates cellular behaviors: cytoskeleton is deformed, cell tentacles are significantly shortened, and cell migration is promoted.


Evidence that formulations of the selective MAO-B inhibitor, selegiline, which bypass first-pass metabolism, also inhibit MAO-A in the human brain.
Selegiline (L-deprenyl) is a selective, irreversible inhibitor of monoamine oxidase B (MAO-B) at the conventional dose (10 mg/day oral) that is used in the treatment of Parkinson's disease. However, controlled studies have demonstrated antidepressant activity for high doses of oral selegiline and for transdermal selegiline suggesting that when plasma levels of selegiline are elevated, brain MAO-A might also be inhibited. Zydis selegiline (Zelapar) is an orally disintegrating formulation of selegiline, which is absorbed through the buccal mucosa producing higher plasma levels of selegiline and reduced amphetamine metabolites compared with equal doses of conventional selegiline. Although there is indirect evidence that Zydis selegiline at high doses loses its selectivity for MAO-B, there is no direct evidence that it also inhibits brain MAO-A in humans. We measured brain MAO-A in 18 healthy men after a 28-day treatment with Zydis selegiline (2.5, 5.0, or 10 mg/day) and in 3 subjects receiving the selegiline transdermal system (Emsam patch, 6 mg/day) using positron emission tomography and the MAO-A radiotracer [(11)C]clorgyline. We also measured dopamine transporter (DAT) availability in three subjects from the 10 mg group. The 10 mg Zydis selegiline dose significantly inhibited MAO-A (36.9±19.7%, range 11-70%, p<0.007)) but not DAT; and while Emsam also inhibited MAO-A (33.2±28.9 (range 9-68%) the difference did not reach significance (p=0.10)) presumably because of the small sample size. Our results provide the first direct evidence of brain MAO-A inhibition in humans by formulations of selegiline, which are currently postulated but not verified to target brain MAO-A in addition to MAO-B.


Is meconium passage a risk factor for maternal infection in term pregnancies?
To study the association between meconium and maternal infection. This was a retrospective cohort study of 678 pregnant women. All term deliveries during a 31-month period with meconium passage were included. Each meconium delivery was matched with a consecutive delivery without meconium at the same gestational age. The overall infection rate was 16%, with 13% of the infections directly related to the pregnancy, labor, and delivery. The endometritis rate was 5%, with 7.1% and 3% in the meconium and no-meconium group, respectively. The chorioamnionitis rate was 8.3%, with 9.5% in the meconium and 7.1% in the no-meconium group. Factors found to be associated with overall obstetric infections had the following odds ratios (ORs) and 95% confidence intervals (CIs): meconium (OR 1.8, 95% CI 1.1, 2.8), internal monitoring (OR 3.4, 95% CI 1.9, 5.9), amnioinfusion (OR 2.0, 95% CI 1.3, 3.3), number of vaginal exams (OR 4.5, 95% CI 2.8, 7.1), length of labor (OR 2.8, 95% CI 1.8, 4.4), and cesarean (OR 3.1, 95% CI 1.9, 5.1). Logistic regression analyses revealed the following ORs and 95% CIs: 1) for endometritis-cesarean (OR 4.2, 95% CI 1.9, 8.9), internal monitoring (OR 2.5, 95% CI 1.1, 5.9), and meconium (OR 2.5, 95% CI 1.1, 5.5), and 2) for chorioamnionitis-length of labor greater than 10 hours (OR 2.7, 95% CI 1.4, 5.6), number of vaginal exams greater than seven (OR 3.4, 95% CI 1.7, 6.6), and use of internal monitors (OR 2.5, 95% CI 1.2, 5.3). Meconium passage increases the risk of postpartum endometritis but not chorioamnionitis. Length of labor, internal monitoring, and number of vaginal exams are risk factors for chorioamnionitis.


The preventive effect and mechanisms of adsorbent supplementation in low concentration aflatoxin B1 contaminated diet on subclinical symptom and histological lesions of broilers.
This study aimed to investigate the subclinical symptom and histological lesions of 21-day-old and 42-day-old broilers exposure to low concentration aflatoxin B1 (AFB1), and the preventive effect with adsorbent (Toxo-MX) supplementation. A total of 576 one-day-old Arbor Acres broilers were randomly allotted into 6 treatments 8 replicates and 12 birds per cage, fed with 0 ppb, 60 ppb and 120 ppb AFB1 contamination diet with or without Toxo-MX supplementation. Results showed both 60 ppb and 120 ppb AFB1 contamination significantly reduced growth performance in 21-day-old broilers (P < 0.05), but not in 42-day-old broilers (P > 0.05), however, AFB1 contamination in diet caused a higher feed to gain ratio (P < 0.05). Broilers of 21-day-old exposure to 60 ppb and 120 ppb AFB1 increased mRNA expression of hepatic inflammatory cytokines, and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity (P < 0.05), 42-day-old broilers showed a same change in 120 ppb but not in 60 ppb of AFB1 contamination (P < 0.05). mRNA expressions of clauding-1, Zonula occludens-1 (ZO-1), and occludin decreased, but Bax, Bcl-2, and caspase-3 increased in 21-day-old broilers exposure to 60 ppb and 120 ppb AFB1 (P < 0.05), broilers of 42-day-old resisted on intestinal aflatoxicosis impairment against 60 ppb AFB1 contamination (P < 0.05), but not in 120 ppb (P < 0.05). Toxo-MX supplementation significantly reversed the detrimental effects on growth performance in both age broilers and reduced the accelerated feed to gain ratio caused by AFB1 (P < 0.05). Intestinal mRNA expression of tight junction and apoptotic genes in both age broilers were recovered by Toxo-MX supplementation (P < 0.05). However, Toxo-MX did not restore the accelerated expression of hepatic inflammation cytokines and SOD, GSH-Px in 120ppb AFB1 group (P < 0.05). The data demonstrated that diet supplementation with Toxo-MX reversed the detrimental effect on growth performance and intestine in broilers exposure to 60 ppb and 120 ppb AFB1. However, did not completely recovered hepatic inflammation induced by AFB1.


Inhibitors produced by algae as an ecological factor affecting bacteria in water ecosystems. I. Dependence between phytoplankton and bacteria development.
Studies were conducted on the eutrophic Mikołajskie Lake in the Mazurian Lake District. Over the period of investigation three maxima of the development of phytoplankton were observed: in the spring, summer and autumn. During the algal blooms the total number of bacteria in the lake strongly decreased and was between several and a dozen time smaller than between blooms. The decrease in the total number of bacteria in water and the elimination of gram positive bacteria during the algal blooms is most probably caused by the production by the algae of substances inhibiting bacterial development.


PCR amplification products are of limited use for the study of DNA/protein interaction.
Conventional methods for labeling double-stranded DNA lead to high specific activity. Yet they often alter the target DNA sequence to such an extent as to prevent a meaningful protein/DNA interaction analysis. Therefore we tried to establish a polymerase chain reaction (PCR)-based method which allows radiolabeling to high specific activity and should maintain the protein binding capability of small double stranded DNA fragments. By using PCR it is possible to label double stranded DNA to high specificity, but the protein binding capability of such DNA is drastically reduced.


The co-existence of two growth hormone receptors and their differential expression profiles between female and male tongue sole (Cynoglossus semilaevis).
Growth hormone receptor (Ghr) is a single-transmembrane pass protein which is important in initiating the ability of growth hormone (Gh) to regulate development and somatic growth in vertebrates. In this study, molecular cloning, expression analysis of two different ghr genes (ghr1 and ghr2) in the tongue sole (Cynoglossus semilaevis) was conducted. As a result, the ghr1 and ghr2 cDNA sequences are 2364 bp and 3125 bp, each of which encodes a transmembrane protein of 633 and 561 amino acids (aa), respectively. Besides, the ghr1 gene includes nine exons and eight introns. The sex-specific tissue expression was analyzed by using 14 tissues from females, normal males and extra-large male adults. Both the ghr1 and ghr2 were predominantly expressed in the liver, and the ghr1 expression level in normal males was 1.6 and 1.4 times as much as those in females and extra-large males, while the ghr2 mRNA expression level in normal males was 1.1 and 1.2 times as much as those in females and extra-large males, respectively. Ontogenetic expression analysis at early life stages indicated that the ghr1 and ghr2 mRNAs were detected at all of the 35 sampling points (from oosphere to 410days-old). Furthermore, the sex differences in ghr mRNA expressions were also examined by using a full-sib family of C. semilaevis. Significantly higher levels of ghr1 mRNA were observed in males than in females at most stages of the sampling period (P<0.01). The ghr2 mRNA expression at most stages exhibited a significant sexual difference at each sampling point (P<0.01) without any variation trend related with the sexes during the whole sampling period.


Crosstalk between protective autophagy and NF-κB signal in high glucose-induced podocytes.
Despite a great deal of recent studies focused on the pivotal role of autophagy in maintaining podocyte energy homeostasis, the mechanisms of autophagy in regulating transcriptional factors under high glucose (HG) condition are not fully understood. Here, we evaluated the effect of HG on nuclear factor-kappa B (NF-κB) signaling and autophagic process. The results showed that HG promoted autophagy in podocytes. Bafilomycin A1 (Baf A1) further enhanced this effect, but 3-methyadenine (3-MA) inhibited it. The proautophagic effects of HG manifested in the form of enhanced podocyte expression of light chain 3 (LC3)-II. In these cells, blockade of NF-κB signal by ammonium pyrrolidinethiocarbamate constrained in effectively reducing LC3-II up-regulation and increasing podocyte apoptosis. Furthermore, the autophagy inhibitors, such as Baf A1 and 3-MA, significantly enhanced HG-induced NF-κB activation and increased apoptosis. Thus, we conclude that the accumulation of autophagosomes results from enhancement of the autophagic flux, but not the blockage of autophagosome-lysosome fusion by HG. We also prove that HG-induced apoptosis, autophagy, and NF-κB signal are in a close crosstalk through a yet undetermined mechanism in podocytes.


Experience of decortication for restrictive hemodynamics in adults with congenital heart disease.
We treated four postoperative adults with congenital heart disease with severe restrictive hemodynamics (RH), and performed decortication (DC) with the anticipation of some relief of the RH. The catheterizations before DC showed high central venous, and right and left ventricular end-diastolic pressures with "dip-and-plateau" pressure waveforms in the right and left ventricles. Upon myocardial histopathologic examination, moderate myocardial fibrotic change was demonstrated in two of three cases. DC led to decrease in type B natriuretic peptide levels in all cases, resulting in a decline in the central venous, right and left ventricular end-diastolic pressures in three cases. Successful DC-related relief of RH, dilatation of the ventricles with decline in central and end-diastolic pressures, was observed in only one case. Our limited DC-related hemodynamic improvement indicates a complexity of the severe RH, which may represent a unique intractable heart failure pathophysiology in intractable postoperative adult congenital heart disease.


Frequent mutations of beta-catenin gene in sporadic secreting adrenocortical adenomas.
Molecular alterations remain largely unknown in most sporadic adrenocortical tumours and hyperplasias. In our previous work, we demonstrated the differential expression of several Wnt/beta-catenin signalling-related genes implicated in ACTH-independent macronodular adrenal hyperplasias (AIMAH). To better understand the role of Wnt/beta-catenin signalling in adrenocortical tumours, we performed mutational analysis of the beta-catenin gene. We studied 53 human adrenocortical samples (33 adenomas, 4 carcinomas, 13 AIMAH, 3 ACTH-dependent adrenal hyperplasias) and the human adrenocortical cancer cell line NCI-H295R. All samples were screened for somatic mutations in exons 3 and 5 of the beta-catenin gene. Eleven and six samples were analysed for beta-catenin protein expression by Western blotting and immunohistochemistry, respectively. No mutations were detected in adrenocortical carcinomas, AIMAH and ACTH-dependent hyperplasias. Genetic alterations were found in 5 (15%) out of 33 adenomas: three cortisol-secreting adenomas, one aldosterone-secreting adenoma and one nonfunctional adenoma. Two-point mutations occurred at serine residues of codons 37 and 45 (S37C and S45F). The remaining three tumours contained deletions of 6, 55 and 271 bp. H295R cells carry an activating S45P mutation. Western blot analysis of samples with 55- and 271-bp deletions showed an additional shorter and more intense band representing an accumulation of the mutated form of beta-catenin protein. In addition, cytoplasmic and/or nuclear accumulation of beta-catenin was observed in mutated adenomas by immunohistochemistry. Activating mutations of exon 3 of the beta-catenin gene are frequent in adrenocortical adenomas, and further characterization of the Wnt/beta-catenin signalling pathway should lead to a better understanding of adrenal tumourigenesis.


Analysis of morphogenetic movements in the development of the notum anlage of Drosophila melanogaster.
A comparison of the morphogenetic maps of the notum anlage of Drosophila melanogaster derived from the gynandromorph data and mosaics induced by somatic crossing-over during the first instar larval stage revealed that practically no major morphogenetic movements occur in the development of the anlage between the blastoderm and first instar larval stages and the adult stage. By comparing the morphogenetic map derived from gynandromorphs and the fate map derived from data on the transplantation of fragments of the mature wing imaginal disc, it was observed that no major morphogenetic movements occur in the notum anlage between the stages of the allocation of the disc and the mature disc. The results are consistent with the observations of other authors concerning the larval development of eye-antenna, wing and leg discs.


[COVID-19: retrospective analysis of cardiac surgery capacity in German heart centers].
For documentation of COVID-19 pandemic-related changes in cardiac surgery in Germany, the German Society for Thoracic and Cardiovascular Surgery carried out a structured survey. From 79 German institutions for cardiac surgery, 53 took part in the survey. The cut-off date before the pandemic was taken as 1 January 2020 and 30 April 2021 as the cut-off date for data recruitment. At this point of time, the third wave of the pandemic in Germany had already ebbed away and the return to routine surgery seemed to be foreseeable. On 1 January 2020, in 53 German cardiac surgery departments 800 intensive care beds were allocated. In April 2021, this capacity was reduced by 25.6% to 595 beds. Accordingly, the number of operating rooms for cardiac surgery procedures was reduced from 207 to 152 (-26.9%) during the first two lockdowns. During the COVID-19 pandemic, hospital facilities were restructured: 50.9% of participants delegated medical personnel, and 75.5% had to transfer nursing personnel to special COVID-19 facilities in their hospitals. From the start of the COVID-19 pandemic until the end of the third wave during April 2021, cardiac surgery in Germany changed profoundly, leading to a substantial reduction of intensive care beds and operating rooms for patients requiring cardiac surgery.


A true long horn-a new species of Macronemus Dejean, 1835 (Coleoptera, Cerambycidae, Lamiinae, Acanthoderini) from the Bolivian Andes.
A new species of long-horned beetle, Macronemus waxrasapa sp. nov. from Bolivian Yungas forest, is described and illustrated.


X-ray Absorption and Electron Paramagnetic Resonance Guided Discovery of the Cu-Catalyzed Synthesis of Multiaryl-Substituted Furans from Aryl Styrene and Ketones Using DMSO as the Oxidant.
The first example of DMSO serving not only as a solvent but also as an oxidant to promote the oxidation of Cu(I) to Cu(II) has been demonstrated. X-ray absorption and electron paramagnetic resonance evidence revealed a single-electron redox process where DMSO could oxidize Cu(I) to Cu(II). The novel discovery guided the rational design of copper-catalyzed oxidative cyclization of aryl ketones with styrenes to furans, providing a new method for the synthesis of multiaryl-substituted furans from cheap and readily available starting materials.


Multifunctional Enzyme with Endoglucanase and Alginase/Glucuronan Lyase Activities from Bacterium Cellulophaga lytica.
Cellulophaga lytica is a Gram-negative aerobic bacterium in the genome of which there are many genes encoding polysaccharide degrading enzymes. One of the enzymes named ClGP contains a glycoside hydrolase domain from the GH5 family and a polysaccharide lyase domain from the PL31 family. The enzyme also contains the TAT signaling peptide and the TIGR04183 domain that indicates extracellular nature of the enzyme. Phylogenetic analysis has shown that the enzymes most closely related to ClGP and containing all four domains (TAT, GH5, PL31, TIGR04183) are widespread among bacterial species belonging to the Flavobacteriaceae family. ClGP produced by the recombinant strain of E. coli was purified and characterized. ClGP exhibited activity of endoglucanase (EC 126.96.36.199) and catalyzed hydrolysis of β-D-glucan, carboxymethyl cellulose sodium salt (CMC-Na), and amorphous cellulose, but failed to hydrolyze microcrystalline cellulose and xylan. Products of CMC hydrolysis were cellobiose and cellotriose, whereas β-D-glucan was hydrolyzed to glucose, cellobiose, cellotetraose, and cellopentaose. ClGP was more active against the poly-β-D-mannuronate blocks than against the poly-α-L-glucuronate blocks of alginic acid. This indicates that the enzyme is a polyM lyase (EC 22.214.171.124). ClGP was active against polyglucuronic acid, so it displayed a glucuronan lyase (EC 220.127.116.11) activity. The enzyme had a neutral pH-optimum, was stable in the pH range 6.0-8.0, and displayed moderate thermal stability. ClGP effectively saccharified two species of brown algae, Saccharina latissima and Laminaria digitata, that suggests its potential for use in the production of biofuel from macroalgae.


The application of genome editing technology in fish.
The advent and development of genome editing technology has opened up the possibility of directly targeting and modifying genomic sequences in the field of life sciences with rapid developments occurring in the last decade. As a powerful tool to decipher genome data at the molecular biology level, genome editing technology has made important contributions to elucidating many biological problems. Currently, the three most widely used genome editing technologies include: zinc finger nucleases (ZFN), transcription activator like effector nucleases (TALEN), and clustered regularly interspaced short palindromic repeats (CRISPR). Researchers are still striving to create simpler, more efficient, and accurate techniques, such as engineered base editors and new CRISPR/Cas systems, to improve editing efficiency and reduce off-target rate, as well as a near-PAMless SpCas9 variants to expand the scope of genome editing. As one of the important animal protein sources, fish has significant economic value in aquaculture. In addition, fish is indispensable for research as it serves as the evolutionary link between invertebrates and higher vertebrates. Consequently, genome editing technologies were applied extensively in various fish species for basic functional studies as well as applied research in aquaculture. In this review, we focus on the application of genome editing technologies in fish species detailing growth, gender, and pigmentation traits. In addition, we have focused on the construction of a zebrafish (Danio rerio) disease model and high-throughput screening of functional genes. Finally, we provide some of the future perspectives of this technology.


Scanning and transmission electron-microscopic study of the development of the podocyte in the human fetus.
The metanephric kidneys of seven human fetuses at 11-17 weeks' gestation were examined by scanning and transmission electron microscopy in order to evaluate differentiation of glomerular podocytes. When glomeruli were in the stage of S-shaped bodies, the surface of the visceral epithelium of renal corpuscles was smooth, with indistinct cell borders. As the glomeruli developed, visceral epithelial cells of renal corpuscles became spherical and resembled clusters of grapes in dense aggregation. In this stage, processes and foot processes were simultaneously formed at the base of epithelial cells. As glomeruli further differentiated, visceral epithelial cells of renal corpuscles began to separate from one another and became flat with the development of vascular loops. Processes and foot processes were exposed for the first time in Bowman's space. In this stage, the degree of differentiation of epithelial cells varied widely among various sites of the glomerulus. As glomeruli developed further, projections became more complex and epithelial cells began to show structures similar to those of adult epithelial cells. The adjacent foot processes arose from different cells throughout the period of morphological differentiation.


Augmentation of monocyte-mediated antibody-dependent cellular cytotoxicity by protein synthesis inhibitors: evidence for an endogenous regulatory mechanism.
Protein synthesis inhibitors, cycloheximide and puromycin, were used in cytotoxic assays employing human peripheral blood monocytes as effectors and sheep erythrocytes as target cells. ADCC could be initiated and could also achieve its full lytic activity in the absence of new protein synthesis. Furthermore, an augmentation of ADCC was observed in the presence of protein synthesis inhibitors. This augmentation was due to an increase in the cytotoxic ability of effector cells rather than a change in the lytic susceptibility of the target. Enhanced cytotoxic potential could not be attributed to an increase in the expression of FcRI but could be due to increased availability of antibody for mediating ADCC as a result of reduced numbers of FcRII. Suppression of prostaglandin-E2 release by monocytes was noted in the presence of cycloheximide, possibly as a result of inhibition of synthesis of cyclooxygenase. However, prostaglandin-E2 and other arachidonic acid metabolites did not appear likely to play a role in negatively regulating human monocyte ADCC since neither cytotoxicity nor cycloheximide-induced augmentation was affected by the presence of exogenous prostaglandin-E2 or arachidonic acid. Cycloheximide was found to induce the secretion of superoxide anions by monocytes, but a role for reactive oxygen species in cycloheximide-induced augmentation of ADCC could not be established by experiments involving the use of catalase or superoxide dismutase. These results raise the possibility that a rapidly turning over protein which negatively regulates monocyte-mediated ADCC exists.


Risk Factors for Stroke in HIV-Positive and-Negative Patients in Pretoria, South Africa.
Low- to middle-income countries experience a marked rise in cardiovascular diseases, and have the highest incidence of HIV infection. Stroke data in HIV-positive patients is still scarce. This study compares risk factors and types of stroke between HIV-positive and -negative patients in South Africa. We conducted a cross-sectional study at Kalafong Provincial Tertiary Hospital in Pretoria over a 10-month period. All adult patients presenting with an acute stroke were included. One hundred and forty consecutive patients with stroke were included, 23% were HIV-positive. The average age in the HIV-positive group was 41 years, compared to 61 years in the HIV-negative group (p < 0.01). Ischemic infarcts occurred in 80.7 and 19.3% were hemorrhagic, with no significant difference between the HIV-positive and -negative group (ischemic: 81% vs 80%; hemorrhagic: 19% vs 20%; p = 0.55). Small vessel infarcts occurred more frequently in HIV-positive patients (25% vs 9.3%; p < 0.02). While 78% of HIV-positive patients presented with concomitant infections, these were found in only 23% of HIV-negative patients (P < 0.001). Hypertension (81% vs 37.5%; p = 0.01) and dyslipidemia (62% vs 38%; p = 0.01) were more prevalent in the HIV-negative patients. Confounding variables were gender and age. Although more than half of the HIV-positive patients were on antiretroviral therapy, the majority (62.5%) showed virological non-suppression. HIV infection occurred in almost one-quarter of stroke patients and was seen more in the younger age group. Small vessel ischemic infarcts and underlying infections were more common in HIV-positive patients. The high number of HIV-positive patients with virological non-suppression is concerning and needs to be addressed.


Correction of rotary malunion of the fingers by metacarpal rotational osteotomy.
Metacarpal rotational osteotomy was performed in 12 patients to correct rotary malunion following fractures of the phalanges and metacarpals. All but one patient obtained excellent correction of their preoperative deformities. The procedure is simple and gives predictable results.


Perturbation of the mitochondrial lysine tRNA population by virus-induced transformation or stress of mammalian cells: functional properties and nucleotide sequence of a mitochondrially associated lysine tRNA.
Eleven isoaccepting lysine tRNAs from mammalian sources are demonstrable by RPC-5 chromatography and polyacrylamide gel electrophoresis. The appearance and amounts of these isoacceptors varies with the source and growth state of cells. One isoacceptor, tRNALys6, observed in preparations of tRNA from some virus-transformed cells in culture, has been characterized by determining functional properties, cellular location, and its nucleotide sequence. tRNALys6 responds primarily to the lysine codon AAA, but it is not used efficiently in a wheat germ translational system in vitro. Compared with lysine isoacceptors 1, 2, 4, 5a, and 5, [3H]lysine appears in vivo in tRNALys6 with a delay of about 3 h. This delay may in part be a result of a less functional tRNA, but a compartmented state of tRNALys6 also appears to be important. tRNALys6 is associated with mitoplasts prepared from KA31 fibroblasts. The nucleotide sequence of tRNALys6 was determined by rapid postlabeling procedures involving limited hydrolysis in formamide, 32P-labeling of 5' ends of fragments with polynucleotide kinase, separation of the nested set of fragments in polyacrylamide denaturing gels, release of 5'-labeled nucleotides with RNase T2, and identification of the released nucleotides by chromatography on PEI cellulose. Confirmation of the positions of major nucleotides was done by using limited digestions by RNases of tRNALys6 labeled with 32P on the 3' terminus in a gel readout procedure. The nucleotide sequence of tRNALys6 differs from that of cytoplasmic lysine tRNAs and mammalian mitochondrial lysine tRNAs. It contains U*, an unidentified modified uridine occurring in the anticodon of some mitochondrial tRNAs. tRNALys6 appears to occur in very limited amounts, or not at all, in most cells unless stressed, but when present it is associated with mitochondria, although it is probably coded in the nucleus.


[Hospital financing and information on hospital clients].
The health care system can be characterized, among other facts, by the multiplicity of involved actors (institutions and individuals) acting for final goals in competition, or even in opposition. The principal goal of the economic organization of the health care system consists in applying a regulation to grant the coordination of the great number of decisions taken by decentralized actors in order to introduce them in economic way into a common final goal. The market forces appear unable to realize the necessary regulation in the health care system and centralized planning can lead to lack of effectiveness. For this reason, it's necessary to make use of a decentralized economic regulation; in this view health care financing systems are essential and could positively influence the behaviour of the actors as well as the costs. Information on hospital Casemix represents an important contribution to better transparency of the working of the health care system and to facilitate competition.


Electrophoretic detection of chitinase isoenzymes using the PhastSystem.
Zymographic assays are described for the detection of chitinase isoenzymes following isoelectric focusing. Method 1 used a polyacrylamide overlay gel containing glycol chitin. Following hydrolysis, visualisation of isoenzymes was achieved by fluorescent counter-staining of the gel with Fluorescent Brightener 28. Method 2 used a cellulose acetate membrane overlay which incorporates 4-methylumbelliferyl substrates, hydrolysis of which released a fluorescent product (4-methylumbelliferone). The pIs of chitinase isoenzymes were estimated by using coloured pI markers. A significant time advantage was obtained over previous methods. The method was used to demonstrate chitinase activity in control rat lungs and in rat lungs infected with the pathogen Pneumocystis carinii.


Intensity-modulated radiation therapy and helical tomotherapy: its origin, benefits, and potential applications in veterinary medicine.
Intensity-modulated radiation therapy (IMRT), especially image-guided IMRT as represented by helical tomotherapy, is a novel approach to therapy and is rapidly evolving. Both of these forms of therapy aim to allow targeted radiation delivery to the tumor volume while minimizing dose to the surrounding normal tissues. Adaptive radiation therapy and conformal avoidance are possible with intensity-modulated therapy and helical tomotherapy, which offer opportunities for improved local tumor control, decreased normal tissue toxicity, and improved survival and quality of life. Human and veterinary patients are likely to benefit from the continued development of this radiation delivery technique, and data over the next several years should be crucial in determining its true benefit.


Quantitative gait analysis in mild cognitive impairment, dementia, and cognitively intact individuals: a cross-sectional case-control study.
Cognitive age-related decline is linked to dementia development and gait has been proposed to measure the change in brain function. This study aimed to investigate if spatiotemporal gait variables could be used to differentiate between the three cognitive status groups. Ninety-three older adults were screened and classified into three groups; mild cognitive impairment (MCI) (n = 32), dementia (n = 31), and a cognitively intact (n = 30). Spatiotemporal gait variables were assessed under single- and dual-tasks using an objective platform system. Effects of cognitive status and walking task were analyzed using a two-way ANCOVA. Sub-comparisons for between- and within-group were performed by one-way ANCOVA and Paired t-tests. Area Under the Curve (AUC) of Receiver Operating Characteristics (ROC) was used to discriminate between three groups on gait variables. There were significant effects (P < 0.05) of cognitive status during both single and dual-task walking in several variables between the MCI and dementia and between dementia and cognitively intact groups, while no difference was seen between the MCI and cognitively intact groups. A large differentiation effect between the groups was found for step length, stride length, and gait speed during both conditions of walking. Spatiotemporal gait variables showed discriminative ability between dementia and cognitively intact groups in both single and dual-tasks. This suggests that gait could potentially be used as a clinical differentiation marker for individuals with cognitive problems.


Self-supporting hierarchical PdCu aerogels for enhanced catalytic reduction of 4-nitrophenol.
This work reports a new kind of self-assembled PdCu monolithic aerogels via a mild reduction process, which exhibits highly efficient catalytic reduction activity towards 4-nitrophenol. The enhanced catalytic reduction performance can be contributed the following unique features of PdCu aerogels: 1) the interconnected channels and three-dimensional network provide a platform for accelerating mass transfer during catalysis; 2) metallic aerogels combined with stretching ultrathin nanowires has a large surface area and good crystallinity affording sufficient reactive sites and high atom utilization; 3) the introduction of nonprecious Cu not only drastically cuts down the cost but also attains the excellent catalytic activity due to the bimetallic intrinsic synergetic effect; 4) the self-supporting feature is good for improving the durability of the catalyst. This study pushes a new avenue to develop robust catalysts for heterogeneous catalytic reactions.


Response to dual blockade of epidermal growth factor receptor (EGFR) and cycloxygenase-2 in nonsmall cell lung cancer may be dependent on the EGFR mutational status of the tumor.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have demonstrated clinical benefit in patients with nonsmall cell lung cancer (NSCLC), particularly those with tumors that have EGFR-TK domain mutations. Moreover, the EGFR and cyclooxygenase (COX)-2 pathways are known to enhance the procarcinogenic effects of each other in different tumor types. Therefore, it was hypothesized that tumor EGFR mutation status may influence the effectiveness of simultaneous EGFR and COX-2 inhibition in patients with NSCLC. Three NSCLC cell lines with varying EGFR mutation status and sensitivities to EGFR-TKIs were selected: H3255 (L858R), H1650 (del E746-A750), and H1781 (wild-type EGFR). Cells were treated with erlotinib, gefitinib, or celecoxib alone, and the combination of both EGFR-TKI inhibitors with celecoxib. Cell survival and apoptosis was assessed and correlated with the expression of COX-2, EGFR, pEGFR, Akt, pAkt, expression, and derived prostaglandin E2 (PGE(2)). Celecoxib by itself was found to have no effects on cell growth or apoptosis in any of the cell lines. Erlotinib and gefitinib inhibited cell growth and induced apoptosis in both mutant cell lines and did so in H1781 cells at 10-fold higher concentrations. Celecoxib when added to erlotinib or gefitinib significantly enhanced the antiproliferative and proapoptotic effects in both mutant cell lines but had no additional effects in H1781 cells. Greater down-regulation of COX-2, EGFR, pEGFR, Akt, pAkt, and PGE(2) was found when H3255 cells were treated with the combination compared with any of the single agents alone. The results of the current study demonstrate that the effectiveness of the addition of celecoxib to an EGFR-TKI is significantly greater in NSCLC cells with EGFR mutations, which is likely due to more complete inhibition of both pathways.


Factors associated with utilization of skilled service delivery among women in rural Northern Ghana: a cross sectional study.
Ghana's current Maternal Mortality Ratio (MMR) of 319 per 100,000 live births makes achievement of the Sustainable Development Goal of 70 maternal deaths per 100,000 live births or less by 2030 appear to be illusory. Skilled assistance during childbirth is a critical strategy to reducing maternal mortality, yet the proportion of deliveries taking place within health facilities where such assistance is provided is very low in Ghana, with huge disparity between urban and rural women. To address the gap in skilled attendance in rural Upper East Region, the Ghana Health Service (GHS) in 2005 piloted a program that involved training of Community Health Officers (CHOs) as midwives. This study explored factors associated with skilled delivery services utilization in a predominantly rural district in Ghana. A cross-sectional study, data was collected from a sample of 400 women between the ages of 15 and 49 years who had given birth a year prior to the study. We used frequencies and percentages for descriptive analysis and chi-square (χ 2 ) test for relationship between independents factors and utilization of skilled delivery services. Of the 400 women included in the analysis, 93.3% of them delivered in a health facility. Almost all of the mothers (97.3%) attended or received antenatal care at their last pregnancy with 75.0% of them having four or more ANC visits. The proportion of women who received ANC and utilized skilled delivery services was high (91.5%). Mother's educational attainment, ANC attendance, frequency of ANC visits, satisfaction with ANC services and possession of valid NHIS card significantly associated with utilisation of skilled delivery services. For a predominantly rural district, the percentage of women who deliver within health facilities where skilled assistance is available is very encouraging and a significant stride towards reducing Ghana's overall MMR. Having four or more ANC visits and improving on the quality of care provided has a great potential of improving uptake of skilled delivery services.


Effect of nonsurgical periodontal therapy verses oral hygiene instructions on type 2 diabetes subjects with chronic periodontitis: a randomised clinical trial.
40 subjects with type 2 diabetes and moderate to severe CP were randomly distributed to groups receiving either NSPT or OHI. Periodontal parameters, glycosylated haemoglobin (HbA1c) and high-sensitivity C-reactive protein (hs-CRP) were evaluated at baseline, 2- and 3-months intervals. 40 subjects with type 2 diabetes and moderate to severe CP were randomly distributed to groups receiving either NSPT or OHI. Periodontal parameters, glycosylated haemoglobin (HbA1c) and high-sensitivity C-reactive protein (hs-CRP) were evaluated at baseline, 2- and 3-months intervals. 15 subjects from NSPT group and 17 from OHI group completed the study. The difference in plaque index (PI) between NSPT and OHI groups were significant at 2 months recall (p = 0.013). There was no significant difference between NSPT and OHI group for all other clinical periodontal parameters, HbA1c and CRP levels. At 3 months post-therapy, periodontal parameters improved significantly in both groups with sites with probing pocket depth (PPD) < 4 mm reported as 98 ± 1.8% in NSPT group and 92 ± 14.9% in OHI group. Mean PPD and mean probing attachment loss (PAL) within the NSPT group reduced significantly from baseline (2.56 ± 0.57 mm, 3.35 ± 0.83 mm) to final visit (1.94 ± 0.26 mm, 2.92 ± 0.72 mm) (p = 0.003, p < 0.001). For OHI group, improvements in mean PPD and mean PAL were also seen from baseline (2.29 ± 0.69 mm, 2.79 ± 0.96 mm) to final visit (2.09 ± 0.72 mm, 2.62 ± 0.97 mm) (p < 0.001 for both). Similarly, HbA1c levels decreased in both groups with NSPT group recording statistically significant reduction (p = 0.038). Participants who demonstrated ≥ 50% reduction in PPD showed significant reductions of HbA1c and hs-CRP levels (p = 0.004 and p = 0.012). NSPT significantly reduced PI at 2 months post-therapy as compared to OHI. Both NSPT and OHI demonstrated improvements in other clinical parameters as well as HbA1c and CRP levels. ClinicalTrials.gov: NCT01951547.


Transient visual field loss after COVID-19 vaccination: Experienced by ophthalmologist, case report.
The first reported case of bilateral transient visual field defect, experienced by an ophthalmologist, which developed shortly after COVID-19 vaccination (CoronaVac, Sinovac Biotech Ltd., Beijing, China) and confirmed by computerized automated perimetry. The patient is a 42-year-old Thai ophthalmologist. He developed blurred vision within an hour after the second dose of COVID-19 vaccination. We described his self-observed of sequential symptoms, chronologic events, and management steps. The visual field of left congruous hemianopia with respect to vertical midline was detected with computerized automated perimetry. The possible mechanism could be related to an acute vasospasm of the artery in the postchiasmatic visual pathway, triggered by COVID-19 vaccine, Corona Vac. With increasing use of the COVID-19 vaccine in the near future, it would be prudent for medical staff and ophthalmologists to be considered and aware of this associated condition.


[Comparative studies of calcium resorption modified by vitamin D2 and D3 in patients with chronic renal failure and dialysis patients].
In 16 patients with a chronic renal insufficiency at the stage of the compensated retention and 13 test persons of the chronic haemodialysis programme determinations of the total calcium and of the ionized calcium fraction as well as investigation of the intestinal calcium absorption by means of the isotope 47Ca were performed. In these cases the influence of vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol) on these parameters of the calcium metabolism was compared. Under the treatment with vitamin D a normalization of the serum calcium levels concerning the total calcium concentration as well as with regard to the ionized calcium fraction was achieved; this effect took place parallel to the improvement of the intestinal calcium absorption. Vitamin D3, had a more favourable effect than vitamin D2. The necessary doses are below 40,000 U a day.


Label-free and ultrasensitive colorimetric detection of DNA based on target-triggered quadratic amplification strategy.
Highly sensitive detection of DNA plays a crucial role in biomedical research and clinical diagnosis. Herein, we developed a simple, label-free, isothermal, and ultrasensitive colorimetric method for amplified detection of DNA on the basis of a new quadratic amplification strategy. With the presence of three ingeniously designed hairpin structures and Exonuclease III (Exo III), the target DNA can trigger two independent cycles of reactions: hairpin assembly reaction and Exo III cleavage reaction, which are designed to initiate target DNA recycling amplification and reporter DNA amplification, respectively. Therefore, the proposed method exhibits a high sensitivity toward target DNA with a detection limit of as low as 81 fM, and it can discriminate mismatched DNA from completely matched target DNA. Furthermore, this method could be used as a universal tool for the detection of various DNA sequences and might be further extended for the detection of aptamer-binding molecules.