Phenylpiperazine 5,5-Dimethylhydantoin Derivatives as First Synthetic Inhibitors of Msr(A) Efflux Pump in Staphylococcus epidermidis.
Herein, 15 phenylpiperazine 3-benzyl-5,5-dimethylhydantoin derivatives (1-15) were screened for modulatory activity towards Msr(A) efflux pump present in S. epidermidis bacteria. Synthesis, crystallographic analysis, biological studies in vitro and structure-activity relationship (SAR) analysis were performed. The efflux pump inhibitory (EPI) potency was determined by employing ethidium bromide accumulation assay in both Msr(A) efflux pump overexpressed (K/14/1345) and deficient (ATCC 12228) S. epidermidis strains. The series of compounds was also evaluated for the capacity to reduce the resistance of K/14/1345 strain to erythromycin, a known substrate of Msr(A). The study identified five strong modulators for Msr(A) in S. epidermidis. The 2,4-dichlorobenzyl-hydantoin derivative 9 was found as the most potent EPI, inhibiting the efflux activity in K/14/1345 at a concentration as low as 15.63 µM. Crystallography-supported SAR analysis indicated structural properties that may be responsible for the activity found. This study identified the first synthetic compounds able to inhibit Msr(A) efflux pump transporter in S. epidermidis. Thus, the hydantoin-derived molecules found can be an attractive group in search for antibiotic adjuvants acting via Msr(A) transporter.


Gastrin antibodies: induction, demonstration, and specificity.
Antibodies to the antral hormone gastrin have been induced in the rabbit and detected by passive hemagglutination. Specificity of the antibody, as determined by three methods, is directed to gastrins I and II, gastrin pentapeptide, and gastrin tetrapeptide, as well as to the stage-1 gastrin used for immunization.


Evidence for the existence of a single enzyme catalyzing the phosphorylation of choline and ethanolamine in primate lung.
Choline kinase (ATP:choline phosphotransferase, EC 184.108.40.206) has been isolated and purified 1000-fold from adult African Green monkey lung with a yield of 10%. The purified enzyme also phosphorylated ethanolamine (ratio of ethanolamine kinase to choline kinase = 0.30). This ratio remained constant throughout the purification procedure. The Km for choline (3.0 - 10(-5) M) was lower than that of ethanolamine (1.2 - 10(-3) M.) Choline was also found to inhibit ethanolamine kinase activity by 50% at a concentration of 0.005 mM, while ethanolamine inhibited choline only at very high concentrations (100--150 mM). When the enzyme was subjected to inactivation by heat, hemicholinium-3, trypsin digestion, and p-hydroxymercuribenzoate, both ethanolamine kinase and choline kinase activities were destroyed at the same rate. Freezing and thawing in the absence of glycerol also destroyed both activities at the same rate. Based on these findings, we conclude that in adult African Green monkey lung tissue, there is only one enzyme for the phosphorylation of ethanolamine and choline, and that choline phosphorylation predominates.


A standard to meet high expectations. Assuring quality care for families of intensive care patients.
1. To give truly holistic care, nurses need to appreciate the family stress response to critical care hospitalisation. 2. Standard setting is an ideal way of meeting relatives' self-perceived needs in intensive care. 3. All staff should be involved with implementation for it to be successful.


A comparison of two methods of dietary assessment in peritoneal dialysis patients.
To conduct a comparison of two methods of dietary assessment in patients on peritoneal dialysis. Comparative, cross-sectional study of two methods of dietary assessment (3-day diet diary and 24-hour recall). Data was collected simultaneously by a single experienced dietitian. Each assessment was coded and analyzed blind. Regional speciality peritoneal dialysis training unit. In this study, 30 peritoneal dialysis patients recruited prospectively and consecutively as they attended for out-patient assessment of dialysis adequacy. Age range was 22 to 77 years. Patients were excluded if unwell, younger than 18 years, or had peritonitis. Total energy and protein intakes from both methods were compared. Protein intakes from both methods were compared with the protein catabolic rate generated from urea kinetics. Data obtained from both methods were compared using paired t tests, linear regression, and Bland and Altman techniques. There were no significant differences in the mean daily protein (72.4 g v 76.6 g) and total energy (1757 kcals (7.35 MJ) v. 1897 kcals (7.94 MJ)) intakes determined by the two methods. Positive correlations were seen between the measurements for protein intake (r =.58, P =.0026) energy intake (r =.78, P <.00001), with mean differences of.066 g/kg/d (SD.38) 2.04 kcal/kg/day (SD 6.67), respectively. For both methods there was a similar positive correlation between dietary protein intake and protein catabolic rate. A 24-hour recall was more likely to result in successful collection of data (29 of 30) than 3-day diet diaries (25 of 30) and was less time consuming. These two methods of determining dietary protein and energy intake do not differ significantly in the information they provide. The relative success in obtaining completed records of intake, the shorter time taken and the opportunity for patient education and assessment of other nutrition related factors has led to the adoption of the 24-hour recall method in our institution.


Caring for the caregiver in dementia.
Caregiver stress is higher for dementia caregiving than other types. By focusing on the family, FPs can help to ease the burden.


Response to the "letter to the editor" by Sani Rachman Soleman et al., "spatiotemporal association of low birth weight with Cs-137 deposition at the prefecture level in Japan after the Fukushima nuclear power plant accidents".
We thank Sani Rachman Soleman et al. for three specific points of criticism concerning our investigation of the ecological association between low birth weight (LBW) and radioactive contamination in Japan after the Fukushima Daiichi Nuclear Power Plant (FDNPP) accidents: 1. Ecological variables are not justified enough to adjust potential confounding. 2. The spatiotemporal regression model does not consider temporal reduction in radiation dose rate. 3. Dose-response plot between dose rates and odds ratios overestimates R2 and underestimates p-value. This criticism is a good starting point to explain some of the technical backgrounds of our approach in more detail.


Adjuvant Radioactive iodine 131 ablation in papillary microcarcinoma of thyroid: Saudi Arabian experience [corrected].
Papillary Microcarcinoma (PMC) of thyroid is a rare type of differentiated thyroid cancer (DTC), which according to the World Health Organization measures 1.0 cm or less. The gold standard of treatment of PMC is still controversy. Our aim was to contribute in resolving the debate on the therapeutic choices of the surgical and adjuvant I-131 (RAI) treatment in PMC. From 2000 to 2012, 326 patients were found to have PMC and were retrospectively reviewed for clinicopathological characteristics, treatment outcomes and prognostic factors. Mean age of cohort was 42.6 years (range: 18-76) and the mean tumor size was 0.61 cm ± 0.24; lymph node involvement was seen in 12.9 % of cases. Median follow up period was 8.05 years (1.62-11.4). Total 23 all site recurrences (7.13 %) were observed; more observed in patients without I-131 ablation (p <0.0001). Ten year DFS rates were 89.6 %. Cox regression Model analysis revealed size, histopathologic variants, multifocality, extrathyroidal extension, lymphovascular space invasion, nodal status, and adjuvant RAI ablation the important prognostic factors affecting DFS. Despite excellent DFS rates, a small proportion of patients with PMC develop recurrences after treatment. Adjuvant RAI therapy improves DFS in PMC patients with aggressive histopathologic variants, multifocality, ETE, LVSI, tumor size (> 0.5 cm) and lymph node involvement. Failure of RAI ablation to decrease risk in N1a/b supports prophylactic central neck dissection during thyroidectomy, however more trials are warranted. Adjuvant I-131 ablation following thyroidectomy in PMC patients, particularly with poor prognostic factors improves DFS rates.


Entangled N-photon states for fair and optimal social decision making.
Situations involving competition for resources among entities can be modeled by the competitive multi-armed bandit (CMAB) problem, which relates to social issues such as maximizing the total outcome and achieving the fairest resource repartition among individuals. In these respects, the intrinsic randomness and global properties of quantum states provide ideal tools for obtaining optimal solutions to this problem. Based on the previous study of the CMAB problem in the two-arm, two-player case, this paper presents the theoretical principles necessary to find polarization-entangled N-photon states that can optimize the total resource output while ensuring equality among players. These principles were applied to two-, three-, four-, and five-player cases by using numerical simulations to reproduce realistic configurations and find the best strategies to overcome potential misalignment between the polarization measurement systems of the players. Although a general formula for the N-player case is not presented here, general derivation rules and a verification algorithm are proposed. This report demonstrates the potential usability of quantum states in collective decision making with limited, probabilistic resources, which could serve as a first step toward quantum-based resource allocation systems.


Long-term prognosis of medullary thyroid carcinoma.
The clinical course of patients with medullary thyroid carcinoma (MTC) is variable, even in the subgroup of patients after surgery with curative intent and postoperatively persistent elevated calcitonin levels. This study aimed to evaluate the long-term prognosis of survival in patients with MTC. Long-term survival was analysed in 32 patients with MTC being treated in an endocrine centre over a 40-year period. Patients were classified as having sporadic MTC, familial MTC (FMTC), multiple endocrine neoplasia (MEN) IIA or MEN IIB. Seventeen patients had sporadic MTC (53.1%), eight had MEN IIA (25%) and three had MEN IIB (9.4%); the remaining four patients (12.5%) had not undergone genetic analysis until now. The overall average age at diagnosis was 42.0 years, and the median follow-up time was 9.5 years (range 0.5-39 years). Mortality due to progressive MTC was 15.6%. The 5-year survival rate was 96% (95% CI 89-100), the 10-year survival rate 91% (95% CI 79-100), and the 15-year survival rate 85% (95% CI 78-100). The estimated mean survival time after initial diagnosis was 31 years (95% CI 26.7-37.0). There is a significant difference in survival time between patients achieving complete remission compared with patients with biochemical persistent disease (P = 0.038) or metastasis (P = 0.0003). In five patients, advanced imaging with positron emission tomography/computed tomography (PET/CT) identified additional sites of tumour load. Eight more lymph node metastases were found in four patients and one local tumour recurrence in one patient by PET/CT. The overall prognosis of MTC is favourable, even if the rate of biochemical cure is lower in MTC than in differentiated types of thyroid cancer. This is also true for patients with biochemically persistent disease. Whether the identification of further tumour sites by advanced imaging procedures such as PET/CT translates into a better prognosis in patients with persistently elevated calcitonin levels remains to be investigated.


Bernard, Mitchell, Horner syndrome and others?
Recent findings of parasellar sympathetic connections lead to renewed speculation concerning the ultimate course of the orbital and facial sympathetic pathways.


GISH-based comparative genomic analysis in Urochloa P. Beauv.
The genus Urochloa P. Beauv. [syn. Brachiaria (Trin.) Griseb.] comprises species of great economic relevance as forages. The genomic constitution for the allotetraploid species Urochloa brizantha (cv. Marandu) and Urochloa decumbens (cv. Basilisk) and the diploid Urochloa ruziziensis was previously proposed as BBB1B1, B1B1B2B2 and B2B2, respectively. Evidence indicates U. ruziziensis as the ancestral donor of genome B2 in U. decumbens allotetraploidy, but the origin of the genomes B and B1 is still unknown. There are diploid genotypes of U. brizantha and U. decumbens that may be potential ancestors of the tetraploids. The aim of this study was to determine the genomic constitution and relationships between genotypes of U. brizantha (2x and 4x), U. decumbens (2x and 4x) and U. ruziziensis (2x) via genomic in situ hybridization (GISH). Additionally, chromosome number and genome size were verified for the diploid genotypes. The diploids U. brizantha and U. decumbens presented 2n = 2x = 18 chromosomes and DNA content of 1.79 and 1.44 pg, respectively. The GISH analysis revealed high homology between the diploids U. brizantha and U. decumbens, which suggests relatively short divergence time. The GISH using genomic probes from the diploid accessions on the tetraploid accessions' chromosomes presented similar patterns, highlighting the genome B1 present in both of the tetraploids. Based on GISH results, the genomic constitution was proposed for the diploid genotypes of U. brizantha (B1B1) and U. decumbens (B1'B1') and both were pointed as donors of genome B1 (or B1'), present in the allotetraploid genotypes.


Effect of morphine and morphine-like drugs on carbachol-induced fighting in cats.
In the present experiments, morphine, methadone or pethidine was injected into the cerebral ventricle of the unanesthetized cat after fighting was induced with carbachol injected previously. The fighting evoked by carbachol was sensitive to the depressant action of morphine or pethidine but not to the depressant effect of methadone. The most likely explanation of the depressant effects of the former compounds is that they act on the postsynaptic receptors of central cholinergic neurons.


Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems.
This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to finally obtain rabies VLP in two culture systems: Schott flask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specific rates were quantified over the exponential growth phase and infection stage. The highest uptake specific rate was observed for glucose (42.5 × 10-12 mmol cell/h) in SF and for glutamine (30.8 × 10-12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10-10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The findings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity.


Propolis allergy. (II). The sensitizing properties of 1,1-dimethylallyl caffeic acid ester.
As shown in the preceding paper, propolis or bee-glue is the cause of an increasing number of allergic reactions in persons using it in external preparations and cosmetics. Propolis and its main contact allergen, 1,1-dimethylallyl caffeic acid ester, designated LB-1, show strong sensitizing properties in patients as well as in guinea pig experiments. 9 patients have been patch tested with this compound, 8 of whom reacted strongly. Chemical separation of different propolis samples and poplar bud extracts reveal that LB-1 is always present. Poplar bud secretion is the bee's major source for propolis and hence the origin of LB-1. A warning is indicated, in agreement with several other authors, that propolis should not be used in topical products because of its strong sensitizing properties.


Cephalometric evaluation of facial types in preschool children without sleep-related breathing disorder.
this study was aimed at characterizing the craniofacial structures, i.e. the facial skeleton, cranial base, dentition, pharyngeal airway space, and the hyoid bone position, in healthy preschool children without sleep-related breathing disorder. from lateral cephalometric radiographs taken of 92 children for diagnostic purposes, 45 were selected for the present investigation on the basis of head position, and divided according to the classification of Siriwat and Jarabak (Angle Orthod. 55 (1985) 127) into groups representing counter-clockwise (CC), straight downward (SD), and clockwise (C) facial types. the findings showed that, in comparison with the other groups, CC is associated with larger facial taper and posterior facial height, smaller mandibular line angle, ramus position, lower facial height and cranial base angle. By a similar comparison, C is associated with larger mandibular line, gonial angles and convexity, and with a smaller L-1 to mandibular line angle. There were no significant differences in hyoid bone position among the three groups. However, the distance of the lower pharynx was smaller in CC than in C, while the tongue base of CC was noted to be in a posterior position. thus, the results indicate that there are significant differences in the madibular position and form among the present three groups of children. It is proposed that the objective of vertical facial control ought to be included in the treatment of preschool children with malocclusion and respiratory disorder.


The effects of biochar on soil organic matter pools are not influenced by climate change.
The sustainability of Mediterranean croplands is threatened by climate warming and rainfall reduction. The use of biochar as an amendment represents a tool to store organic carbon (C) in soil. The vulnerability of soil organic C (SOC) to the joint effects of climate change and biochar application needs to be better understood by investigating its main pools. Here, we evaluated the effects of partial rain exclusion (∼30%) and temperature increase (∼2 °C), combined with biochar amendment, on the distribution of soil organic matter (SOM) into particulate organic matter (POM) and the mineral-associated organic matter (MAOM). A set of indices suggested an increase in thermal stability in response to biochar addition in both POM and MAOM fractions. The MAOM fraction, compared to the POM, was particularly enriched in labile substances. Data from micro-Raman spectroscopy suggested that the POM fraction contained biochar particles with a more ordered structure, whereas the structural order decreased in the MAOM fraction, especially after climate manipulation. Crystalline Fe oxides (hematite) and a mix of ferrihydrite and hematite were detected in the POM and in the MAOM fraction, respectively, of the unamended plots under climate manipulation, but not under ambient conditions. Conversely, in the amended soil, climate manipulation did not induce changes in Fe speciation. Our work underlines the importance of discretely taking into account responses of both MAOM and POM to better understand the mechanistic drivers of SOC storage and dynamics.


Organized sports participation and the association with injury in paediatric patients with haemophilia.
With the wide availability of factor and the routine use of prophylaxis boys with haemophilia are now able to participate in regular physical activity, including organized sports. Current guidelines vary regarding specific recommendations for sports participation and concerns remain regarding safety. To determine if participation in organized sports is associated with an increased risk for injury in paediatric subjects with haemophilia. Retrospective single-centre cohort study from January 1, 2008 to December 31, 2010 in male subjects ages 10-18 years with a factor VIII (FVIII) or FIX level <40%. The number of injuries per subject and participation in organized sports was recorded. 48 male subjects with a mean age of 14.3 ± 2.6 years (range: 10-18.8) were included; 64.6% (31/48) FVIII deficiency, 54.2% (26/48) severe haemophilia, 18.8% (9/48) moderate and 27.1% (13/48) mild. The majority [62.5% (30/48)] of subjects participated in at least one season of organized sport. There were 77 injuries in 36/48 (75%) subjects. The mean number of injuries per subject was 1.6 ± 1.5. There was no statistical difference in the mean number of injuries (P = 0.44) or target joint formation (P = 0.52) between the subjects who participated in organized sports compared to those who did not. In this study, participation in organized sports by boys with haemophilia, ages 10-18 years, is common and not associated with an increased number of injuries or the development of a target joint. As injuries occurred equally in both groups, concerted efforts should be directed at reducing injuries in all patients.


New Role of Bioethics in Emergency Situations on the Example of COVID-19.
The COVID-19 pandemic has dramatically changed existing norm perception both in medical practice and in society. In the context of limited resources and absence of a known treatment protocol, this change particularly affected healthcare system, doctors, and the distribution of a life-support therapy. One of the main bioethical dilemmas presented by coronavirus epidemic is the discrepancy between public health ethics, characterized by the fair distribution of limited resources and public safety orientation, and patient-specific clinical ethics. The COVID-19 pandemic forced health workers into tragic situations that they had never previously experienced, when lack of available health care workers, ventilators, or hospital beds creates a necessity to classify and prioritize patients to determine who will get (or will not get) what type of care and where. Another important issue is the digital control of citizens who must reduce their freedom for the sake of the health of other citizens. This caused great concern among many people, who fear that the current situation will lead to their manipulation in the future. There is also a question of responsibility among politicians and authorized organizations for the health of not only the population of their country, but also the entire planet. These and other current issues require bioethical expertise.


Blowing Off Steam: Virus Inhibition of cGAS DNA Sensing.
Detection of viral DNA is essential for eliciting mammalian innate immunity. However, viruses have acquired effective mechanisms for blocking host defense. Indeed, in this issue of Cell Host & Microbe, Wu et al. (2015) discover a herpesviral strategy for inhibiting the prominent host sensor of viral DNA, cGAS.


The diminution of Heterochromatic chromosomal segments in Cyclops (Crustacea, Copepoda).
The chromosomes of Cyclops divulsus, C. furcifer, and C. strenuus, like those of several other Copepods, undergo a striking diminution of chromatin early in embryogenesis. The process is restricted to the presumptive soma cells and occurs at the 5th cleavage in C. divulsus, at the 6th and 7th in C. furcifer, and at the 4th in C. strenus. The eliminated chromatin derives from the excision of heterochromatic chromosome segments (H-segments). Their chromosomal location is different in the three investigated species: Whereas in C. divulsus and C. furcifer the H-segments form large blocks-exclusively terminal in the former and terminal as well as kinetochoric in the latter-the germ line heterochromatin in C. strenuus is scattered all along the chromosomes. Extensive polymorphism exists with respect to the length of the terminal H-segments in C. furcifer, and with respect to the overall content of heterochromatin in the chromosomes of C. strenuus. In a local race of C. strenuus an extreme form of dimorphism has been found which is sex limited: females as a fule are heterozygous for an entire set of large (heterochromatin-rich), and a second set of small chromosomes in their germ line. Males are homozygous for the large set. In the first three cleavage divisions the H-polymorphism is solely expressed through differences of chromosome length. Following diminution the differences between homologous have disappeared. Feulgen cytophotometry demonstrates that in the three species the 1C DNA value for the germ line, as measured in sperm, is about twice that measured in somatic mitoses (germ line/soma C-values in picograms of DNA: C. strenuus 2.2/0.9, C. furcifer 2.9/1.44, C. divulsus 3.1/1.8). - The data imply that chromatin diminution is based on a mechanism which allows specific DNA segments, regardless of their location and size, to be cut out from the chromosomes without affecting the structural continuity of the remaining DNA. The mechanism may be analogous to that of prokaryotic DNA excision.


Geographical variation in the management of urolithiasis in Australia.
To examine any geographical variation in the management of urolithiasis amongst the Australian states and territories. Retrospective study with data obtained from the Medicare Australia database, the Royal Australasian College of Surgeons and the Urological Society of Australia and New Zealand. Minimally invasive stone treatment with shock wave lithotripsy (SWL) and ureteroscopy (URS) accounted for the majority of stone treatments in Australia (98%). Variation of stone treatment modalities exists amongst the Australian states and territories with an inverse relationship between the use of SWL and URS. We compared Western Australia (WA) and Queensland (QLD) which have a comparable geographical area. SWL accounts for 1% and 22% of stone treatments in WA and QLD, respectively. In WA, urologists are concentrated in two cities with no SWL available in the private sector. In QLD, urologists are distributed in 11 cities with SWL available in both the public and private sector. The three largest states or territories by geographical area - the Northern Territory, WA and QLD - have stone treatment rates of 1:1337, 1:1110 and 1:2432 per capita of privately insured patients, respectively. In comparison, smaller Australian states/territories such as Tasmania and Victoria have stone treatment rates of 1:619 and 1:765 per capita of privately insured patients, respectively. The distribution of urologists and treatment modalities available in each state or territory appear to play a contributory role in choice of treatment modality. In addition, inequality to stone treatment access exists in geographically large Australian states/territories.


Comparative analysis of trace metal levels in the crab Dotilla fenestrata, sediments and water in Durban Bay harbour, Richards Bay harbour and Mlalazi estuary, Kwazulu-Natal, South Africa.
Durban Bay and Richards Bay Harbours are the largest and most economically active shipping harbours in South Africa supporting a diversity of ecosystems and biota of ecological importance. This study assessed and compared levels of metals in selected tissues of the sand bubbler crab (Dotilla fenestrata), water and sediments from anthropogenically impacted Durban and Richards Bay Harbours with those of Mlalazi estuary, a considered pristine site due to its sheltered catchments. Metal concentrations (Cd, Cu, Pb and Zn) were investigated using Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). Metals concentrations in crab tissues were as follows; exoskeleton > gill > digestive gland, metal concentrations in tissues followed the order Cu > Zn > Pb > Cd. Metal levels in crabs from Durban Harbour (Cd 0.42, Cu 83.8, Pb 2.43and Zn 6.4 μg/g) were significantly higher than Richards Bay (Cd 0.22, Cu 27.7, Pb 1.23 and Zn 9.54 μg/g) and Mlalazi estuary (Cd 0.17, Cu 18.7, Pb 3.53 and Zn 6.91 μg/g). Metal levels in sediments followed the order Zn > Cu > Pb > Cd. Mlalazi had significantly elevated metal levels in sediment (Cd 6.83, Cu 35.63, Pb 33.43 and Zn 56.27 μg/g) compared to Durban Harbour (Cd 2.73, Cu 16.07, Pb 12.20 and Zn 38.70 μg/g) and Richards Bay (Cd 3.10, Cu 16.00, Pb 11.43 and Zn 26.07 μg/g). Metal concentrations in water were Cu > Zn > Pb > Cd save Mlalazi estuary with significantly higher Zn levels. Mlalazi estuary, the considered pristine site had significantly elevated metal levels in the sediments and water compared with the impacted sites; however, had significantly lower metal levels in the crabs' tissues due to lower metal bioavailability. Factors such as high natural metal concentrations, metal speciation, sediment grain and organic matter content could possibly account for high metal concentrations without corresponding bioaccumulation and magnification in crabs from Mlalazi estuary.


Double loop figure-of-8 reentry as the mechanism of multiple atrioventricular node reentry tachycardias.
Seven patients with multiple atrioventricular node reentry tachycardia were analyzed to unravel the mechanism of these tachycardias. Six of the seven patients showed anterograde dual atrioventricular node pathways and one showed anterograde conduction through the fast pathway. Three types of retrograde pathways were noted among these seven patients: (1) the fast pathway with the earliest atrial activation at the His bundle area; (2) the intermediate pathway with the earliest atrial activation at the ostium of the coronary sinus; and (3) the slow pathway with the earliest atrial activation at the ostium of the coronary sinus. All seven patients used the intermediate pathway for retrograde conduction. However, one patient showed evidence of retrograde slow pathway conduction with demonstrable retrograde dual pathways, and another showed evidence of retrograde fast pathway conduction with a shift of atrial activation sequence when conduction switched to the intermediate pathway. Four different types of reentry circuits using either the fast or the slow pathway as the anterograde limb and one of the three retrograde pathways as the retrograde limb were demonstrated in these seven patients, resulting in two types of tachycardias in four patients and three types of tachycardias in three patients. A change in tachycardia type could be induced with atrial or ventricular stimulation. A radiofrequency current delivered to the inferior aspect of Koch's triangle along the tricuspid anulus in five patients resulted in selective ablation or modification of the intermediate pathway or the slow pathway, with preservation of anterograde atrioventricular conduction and abolition of tachycardias. The findings suggest that a double loop figure-of-8 reentry circuit including a fast pathway, a slow pathway, and an intermediate pathway is responsible for multiple atrioventricular node reentry tachycardias.


Effect of hypothyroidism on the purinergic responses of corpus cavernosal smooth muscle in rabbits.
Several studies have reported evidence of hormonal abnormalities in 25-35% of impotent men. Hypothyroidism has been reported to occur in 6% of impotent men. In the present study, we examined purinergic relaxation responses in hypothyroidism in an experimental rabbit model and compared them with controls to evaluate the possible involvement of the purinergic pathway. The study comprised 20 male New Zealand white rabbits. The rabbits were divided into two equal groups. We tested the effects of ATP, alpha beta ATP, and adenosine precontracted with phenylephrine on the isolated corpus cavernosum preparations from control and hypothyroid rabbits. We also evaluated the effects of ATP, alpha beta ATP, and adenosine on the cGMP levels in the isolated corpus cavernosum preparations from control and hypothyroid rabbits. T3, T4, and testosterone levels were significantly lower in hypothyroid rabbits. ATP, alpha beta ATP, carbachol, and electrical field stimulation (EFS)-induced frequency-dependent relaxation responses in the isolated rabbit corpus cavernosum strips precontracted with phenylephrine reduced significantly (P<0.05). Adenosine-induced relaxation responses did not change significantly in hypothyroid rabbits. Reduction of relaxation response in hypothyroid rabbits corpus cavernosum can depend on a decreased release of nitric oxide (NO) from nitrergic nerves and endothelium.


Enzymatic Characteristics and Activities of Gingipains from Porphyromonas gingivalis.
Porphyromonas gingivalis is a gram-negative, rod-shaped, nonmotile bacterium belonging to the phylum Bacteroidetes. It produces abundant amounts of proteases in both cell-associated and secretory forms, including a group of cysteine proteases referred to as gingipains, which have attracted much attention due to their high proteolytic activity associated with pathogenicity. Gingipains are grouped into arginine (R)-specific (RgpA and RgpB) and lysine (K)-specific (Kgp) types. Both Rgp (collective term for RgpA and RgpB) and Kgp gingipains play crucial roles in the virulence of P. gingivalis, including the degradation of host periodontal tissues, disruption of host defense mechanisms, and loss of viability in host cells, such as fibroblasts and endothelial cells. In addition to their function in virulence, gingipains are also essential for the growth and survival of P. gingivalis in periodontal pockets through the acquisition of amino acids and heme groups. Furthermore, Rgp and Kgp gingipains are critical in processing fimbriae and several bacterial proteins that contribute to hemagglutination, coaggregation, and hemoglobin binding. This chapter describes the methods used to analyze gingipains.


Driving under the influence of prescription opioids: Self-reported prevalence and association with collision risk in a large Canadian jurisdiction.
Motor vehicle collisions are an important contributor to prescription opioid use-related morbidity and mortality. The purpose of the current study was to estimate the prevalence of driving under the influence of prescription opioids (DUIPO) in Ontario, Canada, and to measure the association between this behaviour and the risk of a motor vehicle collision. Data were based on telephone interviews with 7857 respondents who reported having driven in the past year. Data were derived from the 2011-2016 cycles of the CAMH Monitor, an ongoing cross-sectional representative survey of adults aged 18 years and older. A binary logistic regression analysis of collision involvement in the previous 12 months was conducted and included demographic characteristics (sex, age, marital status, education, income, region), driving exposure, poor mental health, non-medical use of prescription opioids, and driving after use of alcohol. The prevalence of past-year DUIPO was 3.1%. Controlling for demographic characteristics, driving exposure, and other risk factors, self-reported DUIPO significantly increased the odds of a collision (AdjOR = 1.97; 95% CI 1.08, 3.60; p = 0.026). Based on these findings, DUIPO is a notable road safety issue. Research focused on better understanding the impact of prescription opioids on driver behaviour, reducing the prevalence of DUIPO, and improving drug-impaired driving policy and interventions should be prioritized in public health strategies.


A one-step ligation system for rapid generation of armed, conditionally-replicating adenoviruses.
Conditionally-replicating adenoviruses (CRAd) are becoming a promising strategy for cancer therapy. However, the generation of CRAd is complicated and time-consuming, especially for armed CRAd Ad5-D24. A one-step ligation system is now reported for rapid construction of an armed CRAd Ad5-D24 carrying two foreign transgenes. The system has the following features: (1) the CRAd backbone pCRAd5-D24-BS-Lacza contains two unique restriction sites between the fiber and E4 for easy introduction of a transgene expression cassette; and (2) a bi-directional promoter in pshuttle-BS-Dual allows for the expression of two independent transgenes. As proof of principle, Ad5-D24 CRAd carrying eGFP and mCherry was generated and examined. This novel ligation system will be a versatile and efficient tool in cancer viral gene therapy.


Genetic aberrations in tissue inhibitor of metalloproteinases-3 lead to manifestations of the myofibroblast phenotype in mouse fibroblasts and fibroblasts of patients with Sorsby fundus dystrophy.
Mutations in the C-terminal domain of tissue inhibitor of metalloproteinases-3 (TIMP-3) lead to autosomal dominant hereditary disease of the retina (Sorsby fundus dystrophy). Mouse knock-out and heterozygous knock-in fibroblasts and fibroblasts from patients with Sorsby fundus dystrophy exhibit some characteristics of myofibroblasts. Genetic changes in the timp-3 gene (TIMP-3) lead to a shift towards the myofibroblast phenotype in the fibroblast culture. It is hypothesized that Timp-3 (TIMP-3) plays a role of antitransformation agent preventing myofibroblast transformation in vivo and that the pathogenesis of Sorsby fundus dystrophy is associated with the loss of the antitransformation function by the mutant protein.


[Cortexolone as a modulator of diazepam activity].
The influence of ACTH (200 micrograms/kg), corticosterone (20 mg/kg) and cortexolone (20 mg/kg) on the anxiolytic activity of diazepam was studied. ACTH partly and corticosterone completely blocked the action of diazepam. Cortexolone injection 30 min before the administration of diazepam induced a 100% anxiolytic effect of diazepam in the range of doses from 0.1 to 0.3 mg/kg (ED50 of anxiolytic diazepam effect is 0.2 mg/kg). The role of stress hormones in the regulation of psychotropic drug activity is discussed.


A novel intensive induction therapy for high-risk neuroblastoma utilizing sequential peripheral blood stem cell collection and infusion as hematopoietic support.
To determine the feasibility, toxicities, and the response rate (RR) of a dose intensive, submyeloablative, induction chemotherapy protocol termed EPiC (etoposide, carboplatin, and intensive cyclophosphamide) utilizing sequential peripheral blood stem cell (PBSC) collection and infusion as hematopoietic support in children with newly diagnosed Stage 4 neuroblastoma. Twenty-five children (age >1 year) with Stage 4 neuroblastoma were enrolled. First and third cycles consisted of cyclophosphamide (4 gm/m2) and carboplatin (400 mg/m2). Second and fourth cycles consisted of carboplatin (1 gm/m2), and etoposide (450 mg/m2). PBSC were collected following Cycles 1, 2, and 3 and reinfused in each subsequent cycle. Following EPiC and surgical resection of the primary tumor, patients proceeded to various consolidation therapies. RR was scored using the International Neuroblastoma Response Criteria. Using PBSC infusion following EPiC chemotherapy resulted in a dose intensity averaging 85% of intended dose intensity; and in early neutrophil but not platelet recovery. PBSC were adequately collected in all, but one patient. The protocol had minimal non-hematological toxicities. There was one toxic death. The overall RR was 78%, which included PR (partial response) and VGPR (very good partial response). The 5-year event-free survival and overall survival were 44% and 54%, respectively at a median follow-up of 58.6 months. EPiC is a feasible, well-tolerated, sub-myeloablative, induction chemotherapy protocol for children with high-risk neuroblastoma. RR is equivalent to prior published studies, however, with minimal toxicities. Sequential PBSC collection and infusion is feasible even in very young children.


Complementary and alternative medicine use among older urban African Americans: individual and neighborhood associations.
Older African Americans may be vulnerable to risks due to impaired healthcare access and understudied complementary and alternative medicine (CAM) use and other traditions. We describe CAM use among older African Americans in Baltimore, MD, examining associations among CAM and sociodemographic, health status, healthcare utilization and neighborhood factors. Cross-sectional telephone survey of 95 African Americans ages > or =60 on use of CAM modalities (herb/home remedies, prayer, group spiritual practices, meditation/visualization, massage, chiropractic, acupuncture, relaxation/biofeedback) in prior year. A large majority (88.4%) reported CAM use in the previous year: 50.5% reported CAM use excluding individual prayer. The most commonly reported modalities were individual prayer (96.8%), herbs/home remedies (29.5%) and group spiritual practices (17.0%). Most (77.3%) herbal/home remedy users disclosed use to providers. In multivariable logistic regression at the individual level, CAM was associated with higher number of comorbidities (OR = 1.24, 95% CI: 1.06-1.45), older age (OR = 1.09, 95% CI: 1.01-1.17) and more years of education (OR = 1.24, 95% CI: 1.03-1.49). Adding neighborhood variables attenuated effects of comorbidity (OR = 1.17, 95% CI: 0.99-1.39); residence in more racially integrated neighborhoods (OR = 1.03, 95% CI: 1.00-1.06; p = 0.047) was also important. CAM use was greater than expected. Although disclosure of herb use was high, providers should probe for CAM use.


[Comparative adrenergic activity in the Long Evans and Sprague Dawley rat].
Sprague Dawley rats show a more pronounced reactivity of their alpha and beta receptors than Long Evans rats after an infusion of noradrenaline.


Effect of C.C.C. [2-(chloroethyl)] trimethyl ammonium chloride on growth and sporulation in Fusarium oxysporum f. udum (butl.) Sn. et H.
In Fusarium oxysporum f. udum sporulation of macroconidia was completely inhibited due to the influence of C.C.C. [2-(Chloroethyl)] trimethyl ammonium chloride). Microconidial proliferation, on the other hand, was adversely affected and with increase in concentration of this compound a parallel and proportional decrease in the amount of microconidia was found. Size of microconidia, too, showed a concomitant decline. Although the size of the chlamydospores was also reduced, a tremendous increase in its population was evident, showing a positive response in favour of C.C.C. addition. Fungal mat accumulation, another important aspect of growth, did not evince any inhibitory pattern, compared to the progressive increase in C.C.C. amoung in younger cultures. In older cultures (15-day old), an initial depressing shock of C.C.C. addition could be seen. However, after stepwise and gradual adjustment to the new environment the same positive response of mycelial growth was discernible, although in a less degree than that of the young cultures.


Action potential prolongation exhibits simple dose-dependence for sotalol, but reverse dose-dependence for quinidine and disopyramide: implications for proarrhythmia due to triggered activity.
Plasma drug concentrations in patients who develop torsade de pointes while receiving quinidine or disopyramide treatment have been reported to be usually in or below the therapeutic range, whereas patients developing the same complication during sotalol treatment usually have drug concentrations well above the therapeutic range. We wished to provide a cellular electrophysiologic rationale for this observation. Standard intracellular microelectrode techniques were used to record action potentials (APs) from canine Purkinje fibers at interstimulus intervals (ISI) of 1,000-6,000 ms, with and without three varying concentrations of quinidine, disopyramide, and sotalol. Cesium chloride 0.5 mM was added to reduce spontaneous diastolic depolarization. We observed a biphasic response in action potential duration (APD) to quinidine and disopyramide. Low concentrations tended to prolong APD, particularly at slower drive rates, whereas this effect tended to reverse as the concentration was increased. In contrast, sotalol produced a consistent, monophasic dose-dependent increase in APD across the therapeutic concentration range and well beyond it. We also observed an apparent increased likelihood of early afterdepolarizations (EADs), with or without triggered activity, at low concentrations of quinidine and disopyramide, with a trend toward reversal as the concentration was increased. We conclude that the biphasic dose response observed for APD with quinidine and disopyramide is due to the opposing effects of these agents on outward potassium and inward sodium currents and may cast some light on the clinical observation noted above. Sotalol on the other hand appears to produce EADs and triggered activity only at high concentrations.


Management of United Airway Disease Focused on Patients With Asthma and Chronic Rhinosinusitis With Nasal Polyps: A Systematic Review.
The clinical approach to upper and lower respiratory diseases from a joint perspective, known as united airways disease (UAD), is challenging for health care professionals owing to a paucity of specific studies. This study reviews recent scientific evidence on the management of asthma and chronic rhinosinusitis with nasal polyps (CRSwNP) from a UAD perspective. A systematic search of PubMed, Scopus, and Web of Science was conducted for 9 research questions, and studies published from January 2015 to July 2021 were included. Quality assessment was performed with the Critical Appraisal Skills Programme. In total, 32 publications met the inclusion criteria. Control of type 2 inflammation in UAD (reported in 9 studies) was associated with biologic therapies, for which an impact on asthma, CRSwNP, and/or aspirin/nonsteroidal anti-inflammatory drug-exacerbated respiratory disease outcomes was described in 9 studies. However, there was a lack of scientific evidence on clinical and/or biochemical markers associated with response to biologics in patients with UAD. The benefit on corticosteroid reduction in patients receiving biologics was reported in 9 studies. Three publications reported a positive impact of surgery on asthma and/or CRSwNP outcomes, and the effect of biologics on reducing the need of surgery was consistent across 6 studies. Our results underscore an overall scarcity of scientific evidence on the treatment strategies for these frequent coexisting entities from an UAD approach but also identify several research gaps and unmet needs that should be addressed to ensure optimal diagnosis, management, and follow-up of these patients.


Should forensic autopsies be a source for medical education? A preliminary study.
Practical anatomy sessions including dissection of cadavers are essential for anatomy courses. There are many difficulties in obtaining cadavers. In addition, hardened and discolored cadavers that are fixed with formaldehyde look unrealistic and generate apathy among students. We considered that forensic autopsies may be used as ancillary and supportive practice in anatomy education. We invited the participation of Year 2 medical students in suitable forensic autopsy cases during the course of one year. Specialists of forensic medicine and anatomy provided theoretical support through talks in their specialized fields during the autopsy. At the end of the semester, feedback questionnaire forms were prepared and the students were asked to evaluate these sessions. Forty students participated in the evaluation by completing the questionnaire. Students made positive statements about adequacy of the time of the application, consistency of the structures with theoretical and practical issues shown in anatomy lectures, and necessary explanations of the lecturers during and after the application. We think that forensic autopsies are an attractive supplementary educational model, and we have decided to continue the forensic autopsy practices. We believe that further studies on the evaluation of the sessions using a larger student population will lead to more conclusive results.


Effect of cimetidine, histamine H2 receptor antagonist, in the prevention of experimental stress ulcer in the rat.
The protective effect of cimetidine on gastric mucosal lesions in the rat was investigated. Through prophylactic administration of 3x 25 7g/kh body weight for 24 just prior to the test period, the incidence rate of lesions, as well as upper gastro-intestinal haemorrhage was highly significantly reduced.


Syncytia formation of human transformed cell lines by simian sarcoma virus type I (SSV-I/SSAV-I).
Human cells derived from malignant tumors (HeLa, HEp-2 and KB) and human cells transformed by tumor viruses (KCand RSb) formed syncytia by simian sarcoma virus type I (SSV-I/SSAV-I), but human diploid or non-transformed cells (WI-38, HEL and HEC) did not.


Ontogenical studies on kinetics of lipopolysaccharide-induced response to bromelain-treated mouse erythrocytes in mouse spleen cells. II. Response of spleen cells with or without Fc receptors, C3 receptors, or Ia antigens.
Lipopolysaccharide (LPS)-induced plaque-forming cells secreting IgM (IgM-PFC) and antibodies against bromelain-treated mouse erythrocytes (anti-BrMRBC PFC) on Days 1 and 2 of cultures were quantitatively estimated in spleen cells from mice of various ages. The concentrations of the four groups of PFC changed independently with age. The LPS dose dependency of the PFC response was markedly different between PFC on Days 1 and 2, but not different between anti-BrMRBC PFC and IgM-PFC or between 2- and 10-week-old mice. In a second experiment, spleen cells from 2- and 10-week-old mice were separated into subpopulations with or without Fc receptors, C3 receptors, or Ia antigens, and the LPS-induced PFC responses were quantitatively assessed in each subpopulation. Both the receptor-bearing and -lacking populations included LPS-reactive B cells, and the percentages of the LPS-reactive B cells recovered in the receptor-bearing population increased with age. However, the percentages of anti-BrMRBC PFC recovered in each receptor-bearing or -lacking population were different from those of IgM-PFC. In Ia- populations, the percentages of IgM-PFC on Day 2 were obviously higher than those on Day 1, and both of the percentages increased with age. These results suggest that the four groups of LPS-reactive B cells can be discriminated from each other by their LPS dose dependency and their cell surface markers, and that they develop differently during ontogeny.


Management of oral mucositis in patients with cancer.
Oral mucositis (OM) is a distressing toxic effect of chemotherapy and radiotherapy. It can increase the need for total parenteral nutrition and opioid analgesics, prolong hospital stays, increase the risk of infection, and greatly diminish a patient's quality of life. Nurses play a critical role in the assessment and management of OM. However, nurses face several significant challenges in effectively managing OM. First, the assessment and management of OM is inconsistent across Europe. Second, available treatment options for OM are largely ineffective, aimed at palliation of symptoms, and do not target the underlying pathophysiology. Nursing care has focused primarily on symptom management and pain control. With recent advances in the understanding of the pathobiology of OM and the development of interventions, it is hoped that the quality of care provided to patients will improve dramatically as new agents should allow the goal of OM management to shift from symptom relief to protection and prevention. Nurses should be educated about these novel options for the management of OM.


Effects of nebulised iloprost on pulmonary function and gas exchange in severe pulmonary hypertension.
Nebulised iloprost is established therapy of severe pulmonary hypertension; however, the effects on the bronchoalveolar compartment have not been investigated so far. We studied the short- and long-term effects of nebulised iloprost on pulmonary function tests and gas exchange in 63 patients with severe pulmonary hypertension (idiopathic n=17, chronic thromboembolism n=15, connective tissue disease n=12, congenital heart disease n=11, respiratory diseases n=8). Patients received iloprost in increasing dose up to 140 micro g iloprost/24h via an ultrasonic nebuliser. Short-term effects were assessed before and after every nebulisation: peak expiration flow decreased in mean by 1.9% (423+/-98 to 415+/-98) and percutaneous oxygen saturation increased in mean by 0.7% (90+/-6 to 91+/-5) post-nebulisation. There were no significant differences concerning underlying diagnosis or dose of nebulised iloprost. Within 3 months, 9 patients stopped treatment due to non-compliance with frequent nebulisations (n=3), or severe side effects (n=4); 2 patients with additional obstructive lung disease developed bronchoconstriction. Long-term effects were assessed by pulmonary function tests and gas exchange parameters at baseline and after 3 months treatment. There were no significant differences after 3 months therapy neither in FEV(1), FVC, TLC, residual volume nor in diffusions capacity, SO(2) at rest and during 6 min walking test, also in respect of the underlying diseases. However, there was a significant increase in 6 min walking distance (6 MWD) after 3 months (246+/-113 to 294+/-115 m, P<0.05). In conclusion, treatment with nebulised iloprost leads to functional improvement in severe pulmonary hypertension without systematic adverse short- and long-term effects on pulmonary function test or gas exchange. Patients with additional obstructive lung disease might develop bronchoconstriction. Severe side effects leading to discontinuation of treatment occurred in 9% of patients.


In vitro activities of tigecycline against clinical isolates of Aeromonas, Vibrio, and Salmonella species in Taiwan.
All 198 Salmonella isolates (58.6% of isolates were resistant to tetracycline), 92 Vibrio isolates (4.4% of isolates were resistant to tetracycline), and 200 of 201 Aeromonas isolates (39.3% of isolates were resistant to tetracycline; 1 A. caviae isolate had a tigecycline MIC of 4 mug/ml) in our study were susceptible to tigecycline, by U. S. Food and Drug Administration criteria for Enterobacteriaceae.


An enantioselective fluorimetric assay for alcohol dehydrogenases using albumin-catalyzed beta-elimination of umbelliferone.
3-hydroxybutyl umbelliferyl ethers (R)-1 and (S)-1 are fluorogenic substrates for alcohol dehydrogenases. Their oxidation forms ketone 2, which undergoes beta-elimination of umbelliferone under catalysis by bovine serum albumin, leading to a > 20-fold fluorescence increase at lambda em = 460 +/- 20 nm (lambda ex = 360 +/- 20 nm). Enantioselectivity is determined in two separate tests with each enantiomeric substrate.


Small Molecule Restores Itaconate Sensitivity in Salmonella enterica: A Potential New Approach to Treating Bacterial Infections.
In the context of increasing global antibiotic resistance, the need for alternative therapeutic targets is great. Although new antibiotics and resistance inhibitors provide temporary solutions, they are bound to become obsolete. In this work, we propose a new approach, coined "bacterio-modulation" that aims to restore macrophage potency towards bacterial strains that are able to survive in phagolysosomes. One key defense in the macrophage's arsenal is itaconate, an endogenous molecule with antimicrobial activity. Some intracellular pathogens have evolved to produce itaconate-degrading enzymes, which are required for intracellular proliferation and to promote pathogenicity. We herein present the first molecule able to resensitize Salmonella enterica to itaconate.


Waddington's widget: Hsp90 and the inheritance of acquired characters.
Conrad Waddington published an influential model for evolution in his 1942 paper, Canalization of Development and Inheritance of Acquired Characters. In this classic, albeit controversial, paper, he proposed that an unknown mechanism exists that conceals phenotypic variation until the organism is stressed. Recent studies have proposed that the highly conserved chaperone Hsp90 could function as a "capacitor," or an "adaptively inducible canalizer," that masks silent phenotypic variation of either genetic or epigenetic origin. This review will discuss evidence for, and arguments against, the role of Hsp90 as a capacitor for morphological evolution, and as a key component of what we call "Waddington's widget."


Dynamics of thermographic skin temperature response during squat exercise at two different speeds.
Low intensity resistance training with slow movement and tonic force generation has been shown to create blood flow restriction within muscles that may affect thermoregulation through the skin. We aimed to investigate the influence of two speeds of exercise execution on skin temperature dynamics using infrared thermography. Thirteen active males performed randomly two sessions of squat exercise (normal speed, 1s eccentric/1s concentric phase, 1s; slow speed, 5s eccentric/5s concentric phase, 5s), using ~50% of 1 maximal repetition. Thermal images of ST above muscles quadriceps were recorded at a rate of 0.05Hz before the exercise (to determine basal ST) and for 480s following the initiation of the exercise (to determine the nonsteady-state time course of ST). Results showed that ST changed more slowly during the 5s exercise (p=0.002), whereas the delta (with respect to basal) excursions were similar for the two exercises (p>0.05). In summary, our data provided a detailed nonsteady-state portrait of ST changes following squat exercises executed at two different speeds. These results lay the basis for further investigations entailing the joint use of infrared thermography and Doppler flowmetry to study the events taking place both at the skin and the muscle level during exercises executed at slow speed.


Anti-Leishmania antibodies in blood donors from the Midwest region of Brazil.
The aim of the study was to detect asymptomatic infection by Leishmania sp. in blood donors. Serum samples (430) were tested by Immunofluorescent Antibody Test, and an interview with the blood donors was carried out. Antibodies were detected in 15.6% of samples. The variables associated with the infection were: origin of the donor, presence of builds, parks or squares, sick dog in the neighborhood, and neighboring with leishmaniasis. It was observed association between origin of donors and the presence of sick dog. It is important a careful screening of donors, due to the risk of infection through blood transfusion.


Microenvironmental reprogramming of human dermal papilla cells for hair follicle tissue engineering.
The restoration of hair-inductive potential in human dermal papilla cells (hDPCs) is a tremendous challenge for hair regeneration. Much of the research thus far has indicated that three-dimensional (3-D) culture shows improved efficacy in hair follicle (HF) neogenesis. However, mature HF cannot regenerate in an incomplete microenvironment. This study developed an optimized 3-D co-culture system to restore the hair-inductive characteristics of hDPCs by mimicking the in-vivo microenvironment. As a result, Matrigel-encapsulated hDPCs spontaneously formed into hDPC aggregates (hDPAs), which exhibited better activity, higher proliferation rates, and less apoptosis and hypoxia than the ultra-low attachment culture. Interestingly, the co-culture with the hair matrix cells and dermal sheath cup cells further enhanced the expression of hair regeneration-related genes of hDPAs compared to conditioned medium and improved mature HF induction. In addition, these hDPAs with higher hair inductivity could be produced on a large scale and easily separated for gene expression detection. Finally, the mRNA sequencing, PCR, and WB results showed that the co-culture biomimetic microenvironment stimulated the canonical Wnt signaling pathway and inhibited the BMP signaling pathway. Thus, this co-culture system will provide a reliable platform that allows high-throughput culture, testing, and harvesting of hDPAs for HF tissue engineering. STATEMENT OF SIGNIFICANCE: Extensive hair loss continues to be difficult to treat and causes significant patient morbidity. Hair follicle (HF) tissue engineering may seem to be a way out. However, the absence of the in-vivo microenvironment fails to regenerate mature hairs. This study systematically described a biomimetic co-culture approach to generate better quality human dermal papilla cell aggregates (hDPAs) with improved hair inductive properties, which can be further used for HF tissue engineering. The hDPC microenvironment was reprogrammed through the controllable formation of self-assembled organoids in Matrigel and the tri-culture with hair matrix cells and dermal sheath cup cells. This work indicates that the production of hDPAs could be readily scaled, in theory for large-scale assays, analyses, or therapeutic applications.


Effect of olive-mill waste addition to agricultural soil on the enantioselective behavior of the chiral fungicide metalaxyl.
Certain soil management practices can affect the enantioselective behavior of chiral pesticide enantiomers in agricultural soils. In this work, laboratory experiments were conducted to study the effects of olive-mill waste (OMW) addition to a Mediterranean agricultural soil on the enantioselectivity of sorption, degradation, and leaching processes of the chiral fungicide metalaxyl. Sorption-desorption isotherms indicated that the sorption of metalaxyl enantiomers by unamended and OMW-amended soil (2% w/w) was non-enantioselective and that OMW addition had little effect on the extent of sorption of metalaxyl enantiomers by the soil. Soil incubation experiments revealed that the degradation of metalaxyl in unamended soil was highly enantioselective, with R-metalaxyl being degraded faster (t1/2 = 12 days) than S-metalaxyl (t1/2 = 39 days). OMW addition to the soil increased the half-life of the biologically-active R-metalaxyl enantiomer from 12 to 28 days, and decreased the half-life of the non-active S-metalaxyl enantiomer from 39 to 33 days. Consequently, the enantioselectivity of metalaxyl degradation in the soil was greatly reduced upon OMW addition. Column leaching data were consistent with batch sorption and incubation results, showing similar retardation of S- and R-metalaxyl in unamended and OMW-amended soil and enantioselective leaching of the fungicide only in unamended soil. The results have important implications regarding the biological efficacy and environmental impact of the fungicide when applied as a mixture of enantiomers or racemate to OMW-treated soils.


Accelerated dissipation, soil microbial toxicity and dispersal of antimicrobial resistance in soils repeatedly exposed to tiamulin, tilmicosin and sulfamethoxazole.
The application of manures leads to the contamination of agricultural soils with veterinary antibiotics (VAs). These might exert toxicity on the soil microbiota and threaten environmental quality, and public health. We obtained mechanistic insights about the impact of three VAs, namely, sulfamethoxazole (SMX), tiamulin (TIA) and tilmicosin (TLM), on the abundance of key soil microbial groups, antibiotic resistance genes (ARGs) and class I integron integrases (intl1). In a microcosm study, we repeatedly treated two soils (differing in pH and VA dissipation capacity) with the studied VAs, either directly or via fortified manure. This application scheme resulted in accelerated dissipation of TIA, but not of SMX, and accumulation of TLM. Potential nitrification rates (PNR), and the abundance of ammonia-oxidizing microorganism (AOM) were reduced by SMX and TIA, but not by TLM. VAs strongly impacted the total prokaryotic and AOM communities, whereas manure addition was the main determinant of the fungal and protist communities. SMX stimulated sulfonamide resistance, while manure stimulated ARGs and horizontal gene transfer. Correlations identified opportunistic pathogens like Clostridia, Burkholderia-Caballeronia-Paraburkholderia, and Nocardioides as potential ARG reservoirs in soil. Our results provide unprecedented evidence about the effects of understudied VAs on soil microbiota and highlight risks posed by VA-contaminated manures. ENVIRONMENTAL IMPLICATION: The dispersal of veterinary antibiotics (VAs) through soil manuring enhances antimicrobial resistance (AMR) development and poses a threat to the environment and the public health. We provide insights about the impact of selected VAs on their: (i) microbially-mediated dissipation in soil; (ii) ecotoxicity on the soil microbial communities; (iii) capacity to stimulate AMR. Our results (i) demonstrate the effects of VAs and their application-mode on the bacterial, fungal, and protistan communities, and on the soil ammonia oxidizers; (ii) describe natural attenuation processes against VA dispersal, (iii) depict potential soil microbial AMR reservoirs, essential for the development of risk assessment strategies.


Screening for potential quality markers of Callerya nitida var. hirsutissima. Z.Wei based on components profile, pharmacokinetics, and anti-inflammatory study.
Callerya nitida var. hirsutissima. Z.Wei is a classical, traditional Chinese herbal medicine mostly used to treat rheumatoid arthritis. Recent reports suggest that inconsistent and poor-quality control levels have primarily affected the therapeutic efficacy. Therefore, the aim of the current study was to investigate the active chemical ingredients, stability of components in blood, pharmacokinetics, and pharmacodynamics to specify the potential markers for quality control and quality evaluation of Callerya nitida. The active components in vitro and in vivo were obtained by ultra-high-performance liquid chromatography-mass spectrometry. Moreover, the changes of the bioactive components in the blood were monitored over time (0-24 h) in order to identify stable active components. On this basis, the pharmacokinetic characteristics of these ingredients combined with the anti-inflammatory activity were determined to screen out the potential markers for ensuring the quality control of Callerya nitida. The identified four components, such as calycosin, daidzein, formononetin, and 5-hydroxymethylfurfural, have the characteristics of intrinsic components, clearly defined structures, high exposure values, and in vivo stability, which are important for the therapeutic activity of pharmacologically active materials. Therefore, they can be used as potential markers to control the quality levels of Callerya nitida.


Differences in Anxiety and Depression Among Migrant and Non-Migrant Primary School Children in The Netherlands.
This is the first Dutch study investigating symptoms of five DSM-IV-classified anxiety disorders and depression in a large sample of pre-adolescent children with and without a migration background, adjusting for socioeconomic position (SEP) and social preference. Both are potential explanatory factors for differences in mental health among migrant children. We measured anxiety and depression scores with the self-report Revised Child Anxiety and Depression Scale (RCADS) in 2063 children (aged 8-13 years, 55% girls) in the Netherlands. Surinamese/Antillean, Turkish, and Moroccan children reported significantly higher anxiety scores than Dutch children. SEP and peer rejection partly explained higher anxiety scores. Surinamese/Antillean and Turkish children reported comparable depression scores to Dutch children, but Moroccan children reported lower depression scores after adjusting for SEP and peer rejection. Girls reported higher anxiety and depression levels across all four subgroups. Although differences between children with or without a migration background were small, these may increase in later life as the prevalence of anxiety and depression increases with age.


Spinal cord stimulation in animal models of mononeuropathy: effects on the withdrawal response and the flexor reflex.
Spinal cord stimulation (SCS) is efficacious for pain due to injury of peripheral nerves, and therefore models of mononeuropathy appear to be particularly suitable for an experimental approach to the study of mechanisms underlying the clinical effect of this mode of treatment in chronic neuropathic pain. Virtually all previous experimental studies on SCS have utilized acute and nociceptive types of peripheral pain stimuli to explore the attenuating effects of SCS. In the present study we made use of the two models of supposedly painful neuropathy developed by Bennett and Xie (1988) and Seltzer et al. (1990) to explore the effect of SCS applied with stimulus parameters similar to those used in clinical practice. In rats subjected to ligatures of the sciatic nerve according to these two methods, SCS was applied via chronically implanted electrodes, or acutely via a laminectomy in the lower thoracic region. In awake, freely moving animals SCS produced a marked increase of the withdrawal thresholds to innocuous mechanical stimuli in the form of von Frey filaments. This threshold elevation lasted for up to 40 min after 10 min of SCS. In about one-half of the animals there was also a moderate, but short-lasting increase in the intact leg. The degree and duration of the withdrawal threshold elevation was clearly related to the intensity of SCS which was kept below the level of which a response in the thoracic or leg musculature was produced. In a second series of experiments the effect of SCS, applied acutely via a laminectomy, on the early component (latency: 8-12 msec) of the flexor reflex was studied. As a result of nerve ligation with either of the methods used, the thresholds for evoking the early as well as the late component in the nerve-ligated leg were significantly lower than in the intact one. SCS resulted in a marked and long-lasting increase of the threshold of the early component in the nerve-ligated leg. On the intact side only a slight and short-lasting increase was observed. The late, C fibre-mediated component was not influenced by SCS. The first component of the flexor reflex is conceivably mediated by A beta-fibre activation and it presumably corresponds to the withdrawal response induced by innocuous mechanical stimuli. The lack of effect of SCS on the late reflex component indicates that it selectively influences transmission of A-fibre activity. (ABSTRACT TRUNCATED AT 400 WORDS)


Immunological function of Blimp-1 in dendritic cells and relevance to autoimmune diseases.
Previous studies have identified the immunological functions of transcription factor B lymphocyte-induced maturation protein-1 (Blimp-1) in various adaptive immune cell types such as T and B lymphocytes. More recently, it has been shown that Blimp-1 extends its functional roles to dendritic cells (DCs) and macrophages, two cell types belonging to the innate immune system. The protein acts as a direct and indirect regulator of target genes by recruiting chromatin modification factors and by regulating microRNA expression, respectively. In DCs, Blimp-1 has been identified as one of the components involved in antigen presentation. Genome-wide association studies identified polymorphisms associated with multiple autoimmune diseases such as system lupus erythematosus, rheumatoid arthritis, and inflammatory bowel disease in PRDM1, the gene encoding Blimp-1 protein. In this review, we will discuss the immune regulatory functions of Blimp-1 in DCs with a main focus on the tolerogenic mechanisms of Blimp-1 required to protect against the development of autoimmune diseases.


HtrA family proteases of bacterial pathogens: pros and cons for their therapeutic use.
Because there is an urgent need to develop antibacterial therapies other than antibiotics, research has increasingly focused on the high-temperature-requirement protein A (HtrA) family proteases, which have both serine protease and chaperone activities. The research progresses of the role of HtrA family proteases in the pathogenesis of bacterial infections are summarized, and the pros and cons of exploiting HtrA inhibitors in antibacterial drug development are proposed. A search of PubMed was performed to identify relevant studies. HtrA is essential for bacteria to survive in harsh environments, based on the degradation and refolding of misfolded proteins. Moreover, HtrA family protease can lyse the epithelial cell barrier to promote invasion and can also act as or assist virulence factors to enhance pathogenicity. On the other hand, HtrA secreted by certain bacteria can also affect intra- and interspecies biofilm formation (the mechanism of its promotion or inhibition has not yet been proven). Overall, in view of the role of the HtrA family in promoting bacterial pathogenicity, effective HtrA inhibitors may be an exciting direction for drug development. Therefore, the research progress regarding HtrA inhibitors are summarized and the risks of their application are discussed. This review will be useful both for investigators involved in the HtrA field as well as those wishing to acquire a basic understanding of the role and potential implementations of HtrA.


Cervical spinal brucellosis with epidural abscess causing neurologic deficit with negative serologic tests.
Here we present a case of cervical spinal epidural abscess causing neurologic deficits despite negative serologic tests. Unlike a previously reported case, blood cultures and serologic tests were negative for this 50-year-old male with cervical brucellar spondylodiscitis and neurologic deficits. The diagnosis was made based on magnetic resonance imaging (MRI) findings and tissue culture. We drained the epidural abscess, performed anterior cervical discectomy fusion with an iliac strut bone graft, and administered antimicrobial therapy (gentamicin and doxycycline). At the 1-year follow-up visit, a C-spine lateral plain radiograph revealed solid fusion achieved by autogenous strut bone grafting at C4-C5 and C5-C6, and a T2-weighted image showed that epidural abscess and granulation tissue causing spinal cord compression were absent. In addition, the patient had recovered from all of the neurologic deficits with the exception of voiding difficulty. It is important for clinicians to consider brucellar cervical spondylodiscitis when an epidural abscess is identified in a patient with a history of potential exposure. We recommend both blood and tissue cultures for diagnosis when serologic tests are negative.


Fungal Periprosthetic Knee Joint Infection in a Patient with Metamizole-Induced Agranulocytosis.
We present the case of a 55-year-old female patient with metamizole-induced agranulocytosis after total knee arthroplasty, leading to septic periprosthetic joint infections (PJIs). Owing to metamizole-induced agranulocytosis, the synovial leukocyte count was negative. Here, we discuss the diagnostic challenges evolving from sepsis and neutropenia in patients with suspected PJIs. We suggest an urgent surgical approach, mainly focusing on the clinical presentation preoperatively. Later, our patient developed candidemia and periprosthetic tissue samples were positive for Candida albicans. For fungal PJIs, long-term follow-up studies are lacking and therapeutic recommendations differ. Here, we present our therapeutic approach, including staged revision and 12 weeks of systemic antifungal therapy, and discuss recent findings regarding the therapy of fungal PJIs.


Melatonin Delivery: Transdermal and Transbuccal Evaluation in Different Vehicles.
Melatonin (MLT) could be candidate drug for treatment of several diseases because of its high antioxidant and anticarcinogenic activity and its important biological roles. The aim of this study was to assess the influence of different vehicles on the permeation of MLT through buccal and skin tissues. Formulations were characterized in terms of rheology, drug release and permeation through human skin as well as porcine buccal mucosa. Irradiation experiments were also performed. The lowest amount of MLT released was from oral adhesive paste Orabase® (OB) and the highest from the emulsion system Montanov® 68 (M68). Skin permeation revealed high pattern for Carbopol® 940 (C940) and M68, and poor for poloxamer 407 (P407) and Pluronic® lecithin organogel (PLO). Statistical differences of MLT remaining in skin between M68 vs C940 (p < 0.05) and M68 vs PLO (p < 0.05) were observed. Transmucosal results showed that sodium carboxymethylcellulose (NaCMC) was the best and OB the worst vehicle. P407 and PLO followed similar behaviour. Photostability studies revealed high percentage of degradation of MLT in solution which was also similar when was loaded in OB. The rest of formulations showed low rates of degradation. C940 or M68 and NaCMC can be proposed as formulations for a potential systemic effect of MLT by skin and buccal mucosa routes, respectively. However, if the intended objective is to obtain local action in the skin and buccal mucosa, the proposed formulations are M68 or P407 and PLO.


Comparison of long-term outcomes in a 10-year experience of robotic cystectomy vs. open cystectomy.
To compare the outcomes of robotic-assisted (RARC) vs. open radical cystectomy (ORC) at a single academic institution. We retrospectively identified patients undergoing radical cystectomy for urothelial carcinoma of the bladder at our institution from 2007 to 2017. Data collected included age, sex, Body Mass Index (BMI), Charlson Age-Adjusted Comorbidity Index (CCI), final pathologic stage, surgical margins, lymph-node yield, estimated blood loss (EBL), 90-day complication rate, and length of stay (LOS). We evaluated overall survival (OS) and recurrence-free survival (RFS). Multivariable Cox proportional hazard models were used to adjust for covariates. We identified 232 patients (73 RARC, 159 ORC) who underwent radical cystectomy. Patients who underwent RARC were older (71.8 vs. 67.5, p < 0.05) and had higher CCI scores (6.2 vs. 5.3, p < 0.05). In comparing perioperative outcomes, RARC patients had lower EBL (500 vs. 850, p < 0.01), lower blood transfusion rate (p < 0.01), and lower lymph-node yield (12 vs. 20, p < 0.01), and higher ICU admission rate (29% vs. 16% p < 0.01). There was no difference in BMI (p = 0.93), sex (p = 0.28), final pathological stage (p = 0.35), positive surgical margins (p = 0.47), complications (p = 0.58), or LOS (p = 0.34). Kaplan-Meier analysis showed no difference in OS (p = 0.26) or RFS (p = 0.86). There was no difference in restricted mean survival time for OS (53 vs. 56 months, p = 0.81) or for RFS (65 vs. 64 months, p = 0.90). Cox multivariate regression models showed that surgical approach does not have a significant impact on OS (p = 0.46) or RFS (p = 0.35). Our study indicates that in our 10-year experience, patients undergoing there was no difference between RARC and ORC patients with respect to OS and RFS despite being older and having more comorbidities. Our work supports the importance of patient selection to optimize outcomes.


Rivaroxaban vs. warfarin for the treatment and prevention of venous thromboembolism: A meta-analysis.
Anticoagulant treatment is used to treat and prevent venous thromboembolism (VTE). However, the relative effectiveness of newer anticoagulants vs. warfarin has not been appraised. The aim was to evaluate the safety and efficacy of rivaroxaban for VTE in comparison to warfarin. From January 2000 until October 2021, all related studies were collected by EMBASE, the Cochrane Library, PubMed and Web of Scienceand. During the review process, two reviewers independently analyzed the included studies, including quality evaluation, screening and data extraction. We focused on VTE events as our primary outcomes. In total, 20 trials were retrieved. These studies involved 230,320 patients, of which 74,018 received rivaroxaban and 156,302 received warfarin. Compared with warfarin, the incidence of VTE in rivaroxaban is significantly lower (risk ratio (RR) 0.71, 95% confidence interval (CI) [0.61, 0.84]; P < 0.0001, random effect model), and significantly reduced major [RR: 0.84, 95% CI (0.77, 0.91); P < 0.0001, fixed effect model] and nonmajor [RR: 0.55, 95% CI (0.41, 0.74); P < 0.0001, fixed effect model] bleeding. No significant differences in all-cause mortality between the two groups [RR: 0.68, 95% CI (0.45, 1.02); P = 0.06, fixed effect model]. Rivaroxaban significantly reduced the incidence of VTE compared to warfarin in this meta-analysis. In order to verify these findings, larger sample sizes are required in well-designed studies.


Effect of combined statin and beta-blocker treatment on one-year morbidity and mortality after acute myocardial infarction associated with heart failure.
This retrospective, nonrandomized analysis evaluated the effect of initiating statin or beta-blocker treatment early in the course of heart failure developed during acute myocardial infarction compared with the effect of neither or both treatments. Early initiation of statins or beta blockers alone was associated with improved event-free survival, and the benefits of the combined treatment were additive.


Electrophoretic separation of DNA using a new matrix in uncoated capillaries.
A new separation matrix, consisting of polymer poly(N-isopropylacrylamide) (PNIPAM) and small molecule additive mannitol, was used for double-stranded (ds) DNA and plasmid DNA separation by capillary electrophoresis. The matrix had a low viscosity, which made it very easy to handle. The additive mannitol dramatically enhanced the sieving performance of PNIPAM in TBE buffer. The optimal mannitol concentration 6% in polymer solution, was determined with the consideration of both speed and resolution. A resolution of 0.95 was achieved on the separation of 271/281 bp in the phiX174/HaeIII digest by using 1.5% PNIPAM + 6% mannitol, while the supercoiled, linear and nicked conformers of lambda plasmid were separated in 1% PNIPAM + 6% mannitol, demonstrating the potential use of this new matrix for effective DNA separations. The dramatic impact of mannitol on sieving performance of PNIPAM solution was investigated. pH dependent self-coating ability of PNIPAM was revealed. The presence of mannitol in TBE buffer decreased the pH of the buffer, which led to more efficient self-coating ability of PNIPAM probable due to the formation of hydrogen bonds between PNIPAM molecules and silanol groups at the silica wall.


RNA interference of Xenopus NMDAR NR1 in vitro and in vivo.
Here, we present a short interfering RNA (siRNA) application that reduces the expression of NR1 the obligate subunit of the NMDA receptor (NMDAR) and virtually eliminates NMDAR function in a small subset of neurons within otherwise normally developing Xenopus laevis tadpoles. We designed two plasmids each containing a CMV promoter driving a dsRed "reporter" cDNA and DNA coding for one short hairpin RNA (shRNA) under the control of the U6 promoter. The shRNA was cleaved to produce a siRNA against NR1 transcript (iNR1). NR1 transcript and protein, differentiation and survival of NR1 knockdown neurons, were assayed in vivo. iNR1 effects on NMDAR function used Xenopus tectal neuron cultures, Ca2+ imaging and patch-clamp electrophysiology. NR1 transcript, protein and NMDAR function was significantly reduced or eliminated in iNR1-expressing neurons. Protein, transcript and function for the closely related AMPA receptor (AMPAR) subunit GluR1 was unperturbed. In vivo imaging of small groups of iNR1 neurons at two different ages revealed that siRNA knockdown of the NMDAR does not affect differentiation or survival of young neurons. Thus, with siRNA, molecular hypotheses about synaptic mechanisms of circuit formation can be rapidly tested in intact developing vertebrates using normal neurons in the same animals as controls.


The loss of motorneurons corresponding to specific muscles in the wobbler mutant mouse.
The spinal motorneuronal pools specific to a peripheral muscular territory were studied in the wobbler mutant mouse, with the use of retrograde transport of horseradish peroxidase, which was injected into the musculo-cutaneous nerve and into the distal part of the sciatic nerve. Counts of labeled cells showed that the mean percentage of surviving motorneurons was 48.4% in the case of the musculo-cutaneous nerve and 83.6% in the case of the sciatic nerve. The cell loss was not found to be different in old compared to young mutants, suggesting that motorneuron degeneration is an early event in the course of the disease.


Conformational Ensembles of Antibodies Determine Their Hydrophobicity.
A major challenge in the development of antibody biotherapeutics is their tendency to aggregate. One root cause for aggregation is exposure of hydrophobic surface regions to the solvent. Many current techniques predict the relative aggregation propensity of antibodies via precalculated scales for the hydrophobicity or aggregation propensity of single amino acids. However, those scales cannot describe the nonadditive effects of a residue's surrounding on its hydrophobicity. Therefore, they are inherently limited in their ability to describe the impact of subtle differences in molecular structure on the overall hydrophobicity. Here, we introduce a physics-based approach to describe hydrophobicity in terms of the hydration free energy using grid inhomogeneous solvation theory (GIST). We apply this method to assess the effects of starting structures, conformational sampling, and protonation states on the hydrophobicity of antibodies. Our results reveal that high-quality starting structures, i.e., crystal structures, are crucial for the prediction of hydrophobicity and that conformational sampling can compensate errors introduced by the starting structure. On the other hand, sampling of protonation states only leads to good results when combined with high-quality structures, whereas it can even be detrimental otherwise. We conclude by pointing out that a single static homology model may not be adequate for predicting hydrophobicity.


Cell proliferation patterns at the shoot apical meristem.
The highly stereotypic cell proliferation patterns in many plant species suggest that the strict control of the cell cycle in time and space is an essential basis for ordered development. At the same time, conflicting evidence contradicts this view, indicating that cell division simply follows growth patterns that are dictated by the local availability of nutrients. Recent evidence shows that there is no strict hierarchical relationship between growth and proliferation. Cell expansion and proliferation, for example, are controlled by the same regulators. Cell proliferation depends on nutrient distribution but, in turn, the use of the same nutrients depends on the activity of cell cycle regulators such as E2F transcription factors.


Inhibition of rat salivary gland adenylate cyclase by glycosaminoglycans and high molecular weight polyanions.
The effects of hyaluronic acid, chondroitin-4 and -6 sulphate, dermatan sulphate, heparin and the sulphated, polysugar dextran sulphate on membrane-associated adenylate cyclase were investigated in a plasma-membrane fraction derived from the glands. Adenylate-cyclase activity was inhibited in a dose-dependent fashion by all. The potency (concentration causing a 50 per cent inhibition (Ki) of adenylate cyclase activity) of each molecule varied with the degree of sulphation of the agent tested. The GTP and Gpp(NH)p activation of adenylate cyclase as well as basal-enzyme activity and of adenylate cyclase by isoproterenol (a beta-adrenergic agonist) were inhibited; however, little effect on hormone binding (assessed by [3H]-dihydroalprenolol binding) was observed at Ki concentrations. Inhibition of NaF-activation of adenylate cyclase was not as pronounced as the other inhibitory effects. The findings suggested that the agents inhibited enzyme activity by action at or near the catalytic site; hence the effect of these molecules on forskolin activation of adenylate cyclase was investigated. The polyanions inhibited forskolin stimulation of the enzyme to the same degree that they had previously inhibited basal, GTP and hormonally-stimulated enzyme activity. Thus, these molecules inhibited adenylate cyclase by an action (perhaps charge-related) that directly effected the stabilization of catalytic site of the enzyme. Thus polyanions of the type tested can directly modulate adenylate-cyclase activity and may serve as potential regulatory molecules for various biologic functions which are cyclic-AMP dependent.


Immunodetection of salivary biomarkers by an optical microfluidic biosensor with polyethylenimine-modified polythiophene-C70 organic photodetectors.
This work reports a novel optical microfluidic biosensor with highly sensitive organic photodetectors (OPDs) for absorbance-based detection of salivary protein biomarkers at the point of care. The compact and miniaturized biosensor has comprised OPDs made of polythiophene-C70 bulk heterojunction for the photoactive layer; whilst a calcium-free cathode interfacial layer, made of linear polyethylenimine, was incorporated to the photodetectors to enhance the low cost. The OPDs realized onto a glass chip were aligned to antibody-functionalized chambers of a poly(methyl methacrylate) microfluidic chip, in where immunogold-silver assays were conducted. The biosensor has detected IL-8, IL-1β and MMP-8 protein in spiked saliva with high detection specificity and short analysis time exhibiting detection limits between 80pgmL-1 and 120pgmL-1. The result for IL-8 was below the clinical established cut-off of 600pgmL-1, which revealed the potential of the biosensor to early detection of oral cancer. The detection limit was also comparable to other previously reported immunosensors performed with bulky instrumentation or using inorganic photodetectors. The optical detection sensitivity of the polythiophene-C70 OPD was enhanced by optimizing the thickness of the photoactive layer and anode interfacial layer prior to the saliva immunoassays. Further, the biosensor was tested with unspiked human saliva samples, and the results of measuring IL-8 and IL-1β were in statistical agreement with those provided by two commercial assays of ELISA. The optical microfluidic biosensor reported hereby offers an attractive and cost-effective tool to diagnostics or screening purposes at the point of care.


In vitro distribution of porphyrin metabolites from 10(-3) M delta-aminolevulinic acid in primary neural tissue cultures.
The distribution of porphyrin metabolites (uro-7-6-5-copro-3-proto-heme) from 10(-3) M delta-aminolevulinic acid (ALA) in neuronal and glial primary cultures was examined by an isotopic technique. In both cell types, 8-9% of the total production (cell + medium) was found in the cells and more than 90% in the medium. In both the cells and the medium the predominant porphyrin fraction was uro + 7 carboxylic. The heme fraction of the cells was also very high. Through the exclusion of ALA-synthetase activity, the "Uroporphyria-like-model" of "hepatic porphyria" was demonstrated.


Ceftriaxone prevents the neurodegeneration and decreased neurogenesis seen in a Parkinson's disease rat model: An immunohistochemical and MRI study.
Manganese-enhanced magnetic resonance imaging (MEMRI) is a widely used technique for detecting neuronal activity in the brain of a living animal. Ceftriaxone (CEF) has been shown to have neuroprotective effects in neurodegenerative diseases. The present study was aimed at clarifying whether, in an 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) rat model, the known CEF-induced neuronal protection was accompanied by neurogenesis and decreased loss of neuronal activity. After MPTP lesioning (day 0), the rats were treated with CEF (100mg/kg/day, i.p.) or saline for 15 days. They were then injected with MnCl2 (40mg/kg, i.p.) on day 13 and underwent a brain MRI scan on day 14, then the brain was taken for histological evaluation on day 15. The results showed that MPTP lesioning resulted in decreased neuronal activity and density in the nigrostriatal dopaminergic (DAergic) system and the hippocampal CA1, CA3, and dentate gyrus (DG) areas and reduced neurogenesis in the DG, but in hyperactivity in the subthalamic nucleus (STN). These neuronal changes were prevented by CEF treatment. Positive correlations between MEMRI R1 values and neuronal density in the hippocampus were evidenced. Neuronal densities in the hippocampus and SNc were positively correlated. In addition, the R1 value of the STN showed a positive correlation with its neuronal activity but showed a negative correlation with the density of DAergic neurons in the SNc. Therefore, MEMRI R1 value may serve as a good indicator for PD severity and the effect of treatment. To our knowledge, this is the first study showing that CEF prevents loss of neuronal activity and neurogenesis in the brain of PD rats. CEF may therefore have clinical potential in the treatment of PD.


Seroepidemiological evidence for an association between Chlamydia pneumoniae and atherosclerosis.
The respiratory pathogen Chlamydia pneumoniae has been implicated in the pathogenesis of coronary heart disease. Evidence for this includes the direct demonstration of the organism in atherosclerotic plaques and the results of seroepidemiological studies. Of the several serological tests available, the microimmunofluorescence (MIF) test is the most widely used and has become a 'gold standard' because of its specificity and selectivity. In spite of differences in serological criteria of infection and interpretation, the findings of various research groups who have used the MIF test have suggested a link between atherosclerotic cardiovascular disease and Chlamydia pneumoniae infection. Further research is now required in accordance with standard criteria and using pre-defined techniques to confirm the role of the organism in coronary heart disease.


Unveiling the extreme environmental radioactivity of cryoconite from a Norwegian glacier.
This study is a first survey of the occurrence of artificial (137Cs, 241Am, 207Bi, Pu isotopes) and natural (210Pb, 228Ac, 214Bi, 40K) radionuclides in Norwegian cryoconite. Cryoconite samples were collected before (12 samples) and after (5 samples) a rainfall event, after which 7 cryoconite holes dissapeared. The concentrations of radionuclides in cryoconite samples from the Blåisen Glacier are compared with data from the Arctic and Alpine glaciers. Cryoconite samples from the studied glacier had extremely high activity concentrations of 137Cs, 241Am, 207Bi and 239+240Pu (up to 25,000 Bq/kg, 58 Bq/kg, 13 Bq/kg and 131 Bq/kg, respectively) and also high concentrations of organic matter (OM), comparing to other Scandinavian and Arctic glaciers, reaching up to ~40% of total mass. The outstandingly high concentrations of 137Cs, 241Am, Pu isotopes, and 207Bi on the Blåisen Glacier are primarily related to bioaccumulation of radionuclides in organic-rich cryoconite and might be enhanced by additional transfers of contamination from the tundra by lemmings during their population peaks. The presumed influence of intense rainfall on radionuclide concentrations in the cryoconite was not confirmed.


Phytocompounds From Edible Oil Seeds Target Hub Genes To Control Breast Cancer.
Breast cancer is one of the most commonly diagnosed cancers in woman which accounts for more than 1 in 10 new cancers in the entire world. The recently found four new potential hub genes that show a strong expression in breast cancer are CCNA2, CCNB1, MAD2L1, and RAD51. Nowadays, food habits and lifestyle of an individual are one of the factors for causing cancers. Consumption of seeds on a regular basis is the key factor for leading a good health. Sesame seeds and Sunflower seeds are few examples of cancer fighting seeds. Sesame (Sesamum indicum) is one of the earliest oil seed plant with various phytocompounds present which include lignans, tocopherols, phenolics, polyunsaturated fatty acids, and phytosterols. Sunflower (Helianthus annuus L.) is primarily harvested as an oil seed plant with various phytocompounds present which include flavonoids, phenolic acids, tocopherols, and vitamin B3. These are the few seeds that help women to prevent and also to fight against Breast cancer with its potential anti-cancer activity. The main objective of the current study is to identify the potential phytocompounds present in the cancer fighting seeds using molecular docking and dynamic simulation approach which can further help pharmaceuticals industries in producing targeted drugs against breast cancer hub genes as well as food industries in producing products combining the potential phytocompounds present in the seeds.


Reliability of perceptual assessment for resonance disorders in patients with cleft palate.
Determine reliabilities of perceptual assessment of resonance disorders in people with cleft palate. A prospective study of inter- and intra-rater reliability of perceptual screening assessment of resonance disorders with 6-scale and 2-scale criteria among 19 speech and language pathologists (SLPs) from 30 connected speech samples of children with cleft palate aged 4-17 years via video recorder when compared to principle investigators. The percentage of agreement of inter-rater ranged from 23.33-100.00 while Kappa's coefficients was 0.08-0.67. For intra-rater reliability, the present study showed the percentage of agreement ranged from 46.67-100, Kappa's coefficient was 0.17-1. It seems that the more number of years experiences as an SLP, the better listening skill. Reliabilities of perceptual assessment of resonance varied widely but it appears that listening skill might be directly related to experiences. Training programs for critical listening skills are needed.