Physiological effect of the alterations of plasmids derived from Neisseria gonorrhoeae.
Transfer of plasmid DNA from penicillinase-producing Neisseria gonorrhoeae to Escherichia coli produced relatively small colonies that became pigmented in the presence of the dye, congo red. Mutations to large, non-pigmented colonies were frequent, and were the result of a variety of alterations in these plasmids. Miniplasmid formation was the major alteration that led to non-pigmented colonies. However, deletions in any of three putative replication regions resulted in non-pigmented colonies. These altered plasmids mediated significantly less beta-lactamase production than that of the parent plasmid. A new origin of replication was discovered and mapped within the miniplasmid. This origin was responsible for: 1) high plasmid copy number, 2) stability of inheritance, and 3) the majority of beta-lactamase produced in culture. The evolution of gonococcal plasmids may be followed in E. coli, since less defective plasmids produce nonpigmented colonies on congo red agar medium.


[Effect of preoperative preparation on the state of the surfactant in chronic pneumonia].
A complex of physico-chemical, biochemical and morphologic methods was used to study the surfactant state in resected lungs of 44 patients with chronic suppurative processes. A possibility of correction of some surface--active properties of the surfactant through the effect of the preoperative management with the use of intrapulmonary electrophoresis of antibiotics, hydrocortisone and heparin was established.


The Influence of AA29504 on GABAA Receptor Ligand Binding Properties and Its Implications on Subtype Selectivity.
The unique pharmacological properties of δ-containing γ-aminobutyric acid type A receptors (δ-GABAARs) make them an attractive target for selective and persistent modulation of neuronal excitability. However, the availability of selective modulators targeting δ-GABAARs remains limited. AA29504 ([2-amino-4-(2,4,6-trimethylbenzylamino)-phenyl]-carbamic acid ethyl ester), an analog of K+ channel opener retigabine, acts as an agonist and a positive allosteric modulator (Ago-PAM) of δ-GABAARs. Based on electrophysiological studies using recombinant receptors, AA29504 was found to be a more potent and effective agonist in δ-GABAARs than in γ2-GABAARs. In comparison, AA29504 positively modulated the activity of recombinant δ-GABAARs more effectively than γ2-GABAARs, with no significant differences in potency. The impact of AA29504's efficacy- and potency-associated GABAAR subtype selectivity on radioligand binding properties remain unexplored. Using [3H]4'-ethynyl-4-n-propylbicycloorthobenzoate ([3H]EBOB) binding assay, we found no difference in the modulatory potency of AA29504 on GABA- and THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol)-induced responses between native forebrain GABAARs of wild type and δ knock-out mice. In recombinant receptors expressed in HEK293 cells, AA29504 showed higher efficacy on δ- than γ2-GABAARs in the GABA-independent displacement of [3H]EBOB binding. Interestingly, AA29504 showed a concentration-dependent stimulation of [3H]muscimol binding to γ2-GABAARs, which was absent in δ-GABAARs. This was explained by AA29504 shifting the low-affinity γ2-GABAAR towards a higher affinity desensitized state, thereby rising new sites capable of binding GABAAR agonists with low nanomolar affinity. Hence, the potential of AA29504 to act as a desensitization-modifying allosteric modulator of γ2-GABAARs deserves further investigation for its promising influence on shaping efficacy, duration and plasticity of GABAAR synaptic responses.


Inhibitory and excitatory effects on respiration by phrenic nerve afferent stimulation in cats.
Respiratory effects of electrical stimulation of phrenic nerve afferents were studied in anesthetized cats, either spontaneously breathing or paralyzed and ventilated. The type of phrenic afferent fibers activated was controlled by recording the evoked action potentials from dorsal root fibers. In both preparations, stimulation at a strength sufficient to activate small diameter myelinated phrenic nerve afferents induced a biphasic response. The first phase lasted a few respiratory cycles and was inhibitory and consisted of a decrease in tidal volume (VT) or phrenic activity (NA), inspiratory time (TI), respiratory duty cycle (TI/Ttot) and instantaneous ventilation (VE) or minute phrenic activity (NMA). Expiratory time (TE) increased and breathing frequency (f) and mean inspiratory flow (VT/TI) or mean inspiratory neural activity (NA/TI) did not change. This short-term response was suppressed in animals pretreated with bicuculline. The second phase was a long-term excitation in which VT or NA, f, VE or NMA and VT/TI increased whereas both TI and TI/Ttot decreased and TE did not change. Unlike published data, our results suggest that small-diameter myelinated phrenic nerve afferents are involved in these responses. These phrenic fibers, like afferents from other muscles, affect respiratory output and may play a role in the control of breathing.


A fast and simple procedure for determination of perfluoroalkyl substances in food and feed: a method verification by an interlaboratory study.
In this study, a simple, fast, and cheap sample preparation procedure for the analysis of three well-known representatives of perfluoroalkyl substances (perfluorooctane sulfonate, perfluorooctanoic acid, and perfluorooctane sulfonamide) was validated in accordance with Commission Decision 2002/657/EC. The method was based on extraction with methanol followed by a dispersive solid phase extraction cleanup step by addition of activated charcoal for fish tissue, fish feed, and milk samples. The novel analytical approach combined with liquid chromatography-tandem mass spectrometry makes it possible to achieve limits of quantification below 1 μg/kg (defined by Commission Recommendation 2010/161/EU). This method provides a high laboratory sample throughput: ten samples in 60 min. The validated procedure was successfully verified in an interlaboratory study.


Production and use of replication-deficient adenovirus for transgene expression in neurons.
Adenoviruses infect a wide range of cell types, do not require integration into the host cell genome, and can be produced as replication-deficient viruses capable of expressing transgenes behind any desired promoter. Thus, they are ideal for use in expressing transgenes in the postmitotic neuron. This chapter describes simplifications in the protocols for making recombinant adenoviruses and their use in expressing transgenes in primary neurons of several different types.


Inhibition of mercury release from forest soil by high atmospheric deposition of Ca²⁺ and SO₄²⁻.
As one of the most important natural mercury (Hg) sources, soil release (emission to the atmosphere or leaching to soil water) depends on various factors, some of which can be affected by atmospheric deposition. We studied the effect of flue gas desulfurization gypsum (FGDG) addition on soil Hg release in a Masson pine (Pinus massoniana) forest in southwestern China. FGDG addition simulated atmospheric deposition of Ca(2+), SO4(2-) and Hg, which are commonly high in China. Results showed that Hg concentration in soil water decreased with the gypsum treatment, suggesting that the mobility of Hg in mineral soil was reduced. Moreover, the application of gypsum also seems to have decreased Hg emission from the soil, shown by the lower Hg contents in leaf tissues of ground vegetation in the treated plots than in the reference. Both Hg mobility in the soil and Hg emission to the atmosphere were decreased despite the additional Hg input from FGDG. The decreased DOC concentration in soil water and the elevated organic sulfur content in the soil Oe & Oa horizons were speculated to result in an enhanced capacity of surface soil to bind Hg, and thus to reduce Hg release from the soil. However, with the increasingly stringent control of particulate matter (PM) and sulfur dioxide (SO2) emissions in China, the deposition of Ca(2+) and SO4(2-) is expected to decrease, and their ability to inhibit soil Hg release is likely to decline in the future.


Biochemical genetics of Fundulus heterolitus (L.). I. Temporal and spatial variation in gene frequencies of Ldh-B, Mdh-A, Gpi-B, and Pgm-A.
Natural populations of Fundulus heteroclitus show extensive spatial variation in gene frequencies at four unlinked polymorphic loci. Large clinal changes in gene frequencies were found for Ldh-B, Mdh-A, and Gpi-B, whereas the spatial variation for the Pgm-B locus was small. Since the geographical area over which these clines are found is characterized by a steep thermal gradient, the clines in gene frequency are correlated with a directional change in mean water temperature. Maximum gene diversity of these four loci was correlated with annual fluctuations in water temperature. Temporal stability of the allelic frequencies was established for a 2--4 year period.


Long-term predictive value of salivary microbial diagnostic tests in children.
To investigate in a group of children (n=183) the predictive value of early colonization of salivary lactobacilli and candida on caries development in primary molars and permanent first molars in 7 years of follow-up time by applying a longitudinal survival analysis method. Longitudinal cohort study of clinical outcomes based on dental records. The Kaplan-Meier survival analysis method was used to compare survival times to caries onset in primary molars and permanent first molars for children colonized by salivary lactobacilli and candida and children who were not colonized by those microbes at baseline when children's mean age was 2.5 years. The survival time was the time elapsing between the birth of a child and the first restoration as a result of dental caries, that is the time when caries has progressed to a stage when a dentist has made a decision to restore the affected tooth. The survival curves for primary and permanent molars were consistently lower for those children whose mouths were colonized by salivary lactobacilli at baseline. A statistically significant relationship was found with onset of caries in both primary and permanent molars and colonization by salivary lactobacilli. The survival of primary second molars was 60% at the age of 9 years in the group colonized by lactobacilli and 80% in those children whose mouths were not colonized. Colonization by candida was also a significant risk factor for caries onset in primary molars but not in permanent first molars. Children whose mouths were colonized by salivary lactobacilli at baseline were more susceptible to caries onset in both primary and permanent molars at any given time during follow- up of seven years than children who were not colonized by those caries associated microorganisms.


Catheter-induced pulmonary artery haemorrhage resulting from a pneumothorax.
We present a case of pulmonary artery perforation in a patient who developed a pneumothorax after cardiac surgery. In the process of inserting a chest tube the patient became tachypnoeic, and developed haemoptysis. The trachea was intubated, and right bronchial intubation was performed with persistent bleeding. The pulmonary artery catheter was gently withdrawn and the balloon inflated, with cessation of bleeding. The patient was taken to the operating room, a bronchial blocker was placed in the right lower lobe bronchi, and the pulmonary artery catheter was removed. The bronchial blocker was removed the following day with no bleeding. The aetiology of perforation was secondary to the pneumothorax, which caused a shift of the mediastinum to the right, elevated pulmonary artery pressures, and the distal migration of the catheter through the pulmonary artery. It is recommended that treatment include tracheal intubation, inflation of the pulmonary artery catheter balloon, and the placement of a right lower lobe bronchial blocker.


Limb fractures in patients with spinal cord injury.
Limb fractures that occur late in patients with spinal paralysis are considered to be different enough to warrant variations in treatment. In the author's experience, treatment ensures that the functional level is unchanged when healing occurs. Treatment includes use of splints, made of soft materials, that are effective, inexpensive, safe, and allow for good healing. There is motion at the fracture site but, in the author's experience, healing occurs in all fractures except those at the femoral neck. Treatment with soft materials is the least expensive treatment method because it does not require hospitalization, surgery is not recommended, the bone is so soft and pathological that it does not hold firmly with internal fixation, rehabilitation is simplified.


Detection efficacy of analog [18F]FDG PET/CT, digital [18F]FDG, and [13N]NH3 PET/CT: a prospective, comparative study of patients with lung adenocarcinoma featuring ground glass nodules.
This prospective study compared the detection efficacy of analog 18F-fluoro-2-deoxyglucose (18F-FDG) positron emission tomography (PET)/computed tomography (CT) (aF PET/CT), digital [18F]FDG PET/CT (dF PET/CT), and digital 13N-ammonia (13N-NH3) PET/CT (dN PET/CT) for patients with lung adenocarcinoma featuring ground glass nodules (GGNs). Eighty-seven patients with lung adenocarcinoma featuring GGNs who underwent dF and dN PET/CT were enrolled. Based on the GGN component, diameter, and solid-part size, 87 corresponding patients examined using aF PET/CT were included, with age, sex, and lesion characteristics closely matched. Images were visually evaluated, and the tumor to background ratio (TBR) was used for semi-quantitative analysis. Ultimately, 40 and 47 patients with pure GGNs (pGGNs) and mixed GGNs (mGGNs), respectively, were included. dF PET/CT revealed more positive lesions and higher tracer uptake in GGNs than did aF PET/CT (53/87 vs. 26/87, p < 0.05; TBR: 3.08 ± 4.85 vs. 1.42 ± 0.93, p < 0.05), especially in mGGNs (44/47 vs. 26/47, p < 0.05; TBR: 4.48 ± 6.17 vs. 1.78 ± 1.16, p < 0.05). However, dN PET/CT detected more positive lesions than did dF PET/CT (71/87 vs. 53/87, p < 0.05), especially in pGGNs (24/40 vs. 9/40, p < 0.05). dF PET/CT provides superior detection efficacy over aF PET/CT for patients with lung adenocarcinoma featuring GGNs, particularly mGGNs. dN PET/CT revealed superior detection efficacy over dF PET/CT, particularly in pGGNs. aF, dF, and dN PET/CT are valuable non-invasive examinations for lung cancer featuring GGNs, with dN PET/CT offering the best detection performance. • Digital PET/CT provides superior detection efficacy over analog PET/CT in patients with lung adenocarcinoma featuring GGNs. • dN PET/CT can offer more help in the early detection of malignant GGN.


Peak plasma anti-Xa levels after first and third doses of enoxaparin in women receiving weight-based thromboprophylaxis following caesarean section: a prospective cohort study.
Women undergoing caesarean section are at higher risk for thromboembolic complications following delivery than other parturients. The aim of this study was to determine whether higher doses of enoxaparin based on body weight are safe and more likely to achieve plasma anti-Xa levels within the accepted thromboprophylactic range. We undertook a prospective cohort study of 80 women undergoing caesarean section in a tertiary obstetric hospital with >6000 deliveries per year. Enoxaparin was administered after caesarean section using the Royal College of Obstetricians and Gynaecologists weight-adjusted dosing guidelines. Plasma anti-Xa levels were measured at baseline and 3-4 h after enoxaparin administration on days one and three postoperatively. The main outcomes of interest were plasma anti-Xa levels and the proportion of patients with plasma anti-Xa levels in the range of 0.2-0.4 IU/mL. The proportion of women with anti-Xa levels between 0.2 and 0.4 IU/mL was 72% (95% CI 60-81%). Unadjusted mean anti-Xa levels were 0.26 ± 0.09 IU/mL and 0.28 ± 0.08 IU/mL on day one and day three respectively. No woman had levels >0.48 IU/mL. The majority of women receiving weight-based enoxaparin thromboprophylaxis following caesarean section achieved plasma anti-Xa levels within the putative thromboprophylactic range. No woman achieved levels associated with an increased risk of bleeding (>0.8 IU/mL). These findings provide a safety basis for a large prospective study using this regimen.


Neddylation promotes protein translocation between the cytoplasm and nucleus.
Neddylation is an ubiquitin-like modification of proteins that affects the activity, stability and protein-protein interaction of its substrates. Apart from its role as a promoter for Cullin ring E3 ligase to positively regulate the ubiquitylation process, other functional studies about neddylation are still lacking. In this study, we developed a system to explore the impact of neddylation on changes in the subcellular localization of proteins at the omics level. By applying a method combining subcellular protein extraction and immunoprecipitation-mass spectrometry (IP-MS), 81 proteins with a tendency to shuttle between the cytoplasm and nucleus due to different neddylation levels were obtained. Among the 81 candidates, transforming growth factor-β (TGF-β)-activated kinase 1 (TAK1) and growth arrest and DNA damage protein 45a (Gadd45a) were confirmed as novel substrates of Nedd8, and neddylation promotes TAK1 nuclear import as well as Gadd45a nuclear export.


Evaluation of a new latex agglutination test for detection of streptococcal antibodies.
A liposome-enhanced latex agglutination test (Leapstrep, Cooper Biomedical, Inc., Malvern, PA) was evaluated for its efficacy as a screening test for antistreptolysin O (ASO) and antideoxyribonuclease B (ADNB) antibodies. A total of 179 sera were tested by both Leapstrep and reference ASO and ADNB test methods. The Leapstrep test had a sensitivity of 77% and a specificity of 69% for ASO antibodies when compared to a microtiter adaptation of the Todd-Hewitt ASO tube test. It had a sensitivity of 76% and 75% specificity for ADNB antibodies as compared to the ADNB microtiter enzyme neutralization test. The overall sensitivity of Leapstrep for detecting ASO and/or ADNB antibodies was 73%. The Leapstrep test failed to detect elevated levels of ASO and/or ADNB antibodies in eight patients with streptococcal infections or poststreptococcal sequelae. The Leapstrep test appears to have insufficient sensitivity to be recommended as a screening test for streptococcal antibodies.


Prevailing patterns and predictor variables in patients with acute tubular necrosis.
The courses of 276 acute tubular necrosis patients referred for dialysis were reviewed in search for prognostic indicators. Sixty-three percent survived. Of 28 possible predictor variables, a posttoxic cause and nonoliguria were favorable, whereas myocardial infarction and peritonitis affected survival unfavorably. Total pareneral nutrition influenced survival favorably only in those with multiple complications or peritonitis. No single variable or combination predicted a lethal outcome. Since survivors were frequently restored to complete health, we advocate an aggressive therapeutic approach even in the face of multiple complications.


The contribution of self-rated health and depressive symptoms to disability severity as a predictor of 10-year mortality in European elderly men.
To investigate the effect of disability severity and the contribution of self-rated health and depressive symptoms to 10-year mortality. Longitudinal data were collected from 1141 men aged 70 to 89 years from the Finland, Italy, and the Netherlands Elderly Study from 1990 to 2000. Disability severity was classified into 4 categories: no disability, instrumental activities, mobility, and basic activities of daily living. Self-rated health and depressive symptoms were classified into 2 and 3 categories, respectively. Multivariate Cox proportional hazard models were used to calculate mortality risks. Men with severe disability had a risk of mortality that was more than 2-fold higher (hazard ratio [HR]=2.41; 95% confidence interval [CI]=1.84, 3.16) than that of men without disability. Men who had severe disability and did not feel healthy had the highest mortality risk (HR = 3.30; 95% CI = 2.52, 4.33). This risk was lower at lower levels of disability and higher levels of self-rated health. The same trend was observed for depressive symptoms. For adequate prognoses on mortality or for developing intervention strategies, not only physical aspects of health but also other health outcomes should be taken into account.


Awareness of population regarding the effects of diabetes on dental implant treatment in Jeddah, Saudi Arabia.
Diabetes mellitus (DM) has several complications. Delayed wound healing, microvascular disease and an impaired response to infections are complications that can have a direct bearing on dental implant therapy. This paper studies the awareness of the population with regard to the effect of DM on dental implant treatment. A validated, close-ended questionnaire was distributed to 506 randomly selected mall-goers in the city of Jeddah. Responses were coded and entered into spreadsheet software (SPSS, IBM). The frequency distribution of the responses was calculated, and inferences were drawn. The study revealed that the majority of the sample were females (62.8%), did not have diabetes (80.4%) and reported a positive family history of diabetes (87.4%). Most of the respondents (56%) believed that uncontrolled diabetes can lead to implant loss and that diabetes affects the healing process (91.6%). Many patients (42%) responded that diabetes could be treated with dental implants if the blood sugar level was controlled. The studied sample revealed a satisfactory level of awareness regarding the association of diabetes and oral hygiene in dental implant therapy. However, there is less than adequate knowledge about the effects of diabetes on dental implants, as the majority of the respondents believe that only controlled diabetics can avail dental implant treatment.


Young Patients With Diabetes Have Decreased Cerebrovascular Reactivity Under Hypercapneic Conditions.
Adult diabetic patients have an abnormal cerebrovascular response to hypercapnia, but there are few studies focused on diabetes mellitus type 1 and cerebral blood flow in pediatric or adolescent patients. We hypothesize that young patients with diabetes exhibit a different response to hypercapnia than normal control counterparts. Using transcranial Doppler techniques, we compared young diabetic patients with healthy controls by measuring cerebral blood velocity before and during carbon dioxide challenge. Subjects with diabetes had decreased cerebral blood velocity reactivity when compared with the control group (P = 0.005). Our results suggest cerebrovascular dysfunction in diabetic patients beginning at an early age. The possibility of long-term implications for cerebrovascular disease demonstrates the need for further studies in the pediatric and adolescent diabetic population to better understand this prevalent condition.


Long-Term Survival of Ischemic Stroke Patients according to Prior Cognitive Status: Dijon Stroke Registry.
Understanding the influence of preexisting cognitive impairment on the poststroke outcome is a critical challenge in the context of current aging and growing population. This study aimed to assess long-term survival of patients with acute ischemic stroke (IS) according to their premorbid cognitive status and to identify contributing factors of death. Patients with IS were prospectively identified among residents of Dijon, France, using a population-based registry (2013-2017). The association between case fatality at 5 years and prestroke cognitive status was assessed by multivariable Cox models adjusted for other clinical characteristics and preexisting brain damage on the initial CT scan including leukoaraiosis, old vascular brain lesions, and cortical and central brain atrophy, as well as major arterial occlusion. 1,049 patients were included (mean age ± SD: 76.3 ± 15.2 years old, 54% women). Case fatality rates at 5 years were 38.1% in patients without cognitive impairment, 65.9% in patients with prior mild cognitive impairment (MCI, n = 132, 12.6%), and 86.6% in patients with dementia (n = 164, 15.6%) (p < 0.001). MCI (HR = 1.39; 95% CI: 1.06-1.81, p = 0.016) and dementia (HR = 1.89; 95% CI: 1.45-2.46, p < 0.001) were both independently associated with higher case fatality after adjustment for clinical variables. The association remained significant after further adjustment for preexisting brain damage and major arterial occlusion (HR = 1.47; 95% CI: 1.10-1.98, p = 0.009, for MCI and HR = 1.90; 95% CI: 1.43-2.53, p < 0.001, for dementia) among patients with available data on the CT scan (n = 916). Factors associated with death were roughly similar across groups. This study highlighted a poor long-term survival of IS patients with preexisting cognitive impairment, independently of other contributing factors of death. It is critical to better understand the trajectory of IS patients with preexisting cognitive impairment and to identify prognostic markers to guide clinicians in their management strategies.


Influence of neoadjuvant chemoradiotherapy on the anal sphincter: ultrastructural damage may be critical.
Neoadjuvant chemoradiotherapy (nCRT) in addition to the curative surgery has been the first of treatment for local advanced rectal cancer because of its benefits in local recurrence and sphincter-saving. However, its side effects on anorectal function have been recognized. The histopathological changes on internal anal sphincter (IAS) have been reported, but ultrastructure changes of external anal sphincter (EAS) are unknown. The aim of this study is to detect the alterations on the gross morphology of IAS and ultrastructure of EAS after nCRT. We collected 34 anal canal specimens of patients undergoing abdominoperineal resection (APR) prospectively. The length and thickness of IAS were measured with vernier caliper. The EAS was dissected for observation with transmission electron microscope (TEM). Ten patients received nCRT (nCRT group) before surgery and 24 underwent APR directly (control group). The length and thickness of IAS in nCRT group were 22.68 ± 3.56 and 5.39 ± 0.74 mm, respectively. These parameters were 21.28 ± 3.62 and 5.35 ± 1.12 mm in control group, respectively. There were no significant differences in the length and thickness of IAS between the two groups (P>0.05). In nCRT group, the sarcomere and myofibril were arranged disorderly and parts of them that were filled with collagenous fiber, triads, and mitochondria were destroyed severely and the glycogenosome also distributed disorderly. Such alterations of EAS did not occur in control group. The nCRT cannot change the gross morphology of IAS, while it induces serious damages to the ultrastructures of EAS which may adversely affect the anorectal function.


A study to validate the colonoscopy simulator.
The aim of this study was to investigate the relation between clinical experience and performance with regard to colonoscopic procedures performed on the HT Immersion Medical Colonoscopy Simulator. The hypothesis is that the performance of novice, intermediate, and experienced operators is different on simulators, just as it is on real patients. 25 Postgraduate doctors were recruited and divided into three groups according to their level of colonoscopic experience. Candidates were asked at random to perform colonoscopy on module 3 or 4 of the HT Immersion Medical Colonoscopy Simulator. Modules 3 and 4 have built-in complex loops, and can demonstrate the candidate's ability either to avoid or undo the loop. Candidates in the first group, termed "novice," each had performed fewer than 10 colonoscopies and included four preregistration house officers (PRHOs), five specialist trainee resistrars (SpRs) and two consultants. This novice group had completed 80 episodes among them. In the second group, termed "intermediate," each candidate had performed between 11 and 100 colonoscopies, and the candidates included five SpRs and two research fellows. This intermediate group had completed 65 episodes. Members of the third group, termed "experienced" each had performed more than 101 colonoscopies, and included one SpR and six consultants. This experienced group had completed 45 episodes between them. A time result of 3,600 s (1 h) was used to denote perforation. The experienced group were shown to perform better than the intermediate group, which in turn performed better than the novice group. The assessment was based on multiple factors including time taken to complete the test, percentage of the colonic mucosa visualized, incidence of colonic perforations, and path length used. The results were highly significant statistically for all these factors ( p < 0.000) except in path length used. This study demonstrated that operators who differ in terms of their clinical experience and technical ability also differ in their performance of simulated colonoscopy. Thus, the findings have shown the simulator technology to be a powerful discriminator of manipulative skills in colonoscopy. The clinical differences that exist between novices and experts in terms of experience and technical expertise in endoscopic procedures are reflected by statistically significant differences in performance on the simulator. It is therefore reasonable to argue that improving proficiency on the simulator may translate into improvements in clinical practice. This study has validated the use of the HT Medical Immersion Colonoscopy Simulator to discriminate among subjects with different levels of experience.


Preparation of an oral thin film containing meclizine hydrochloride: In vitro and in vivo evaluation.
Oral thin film (OTF) is a preparation of postage stamp size, with advantages of flexible, tasty and without water for oral administration. A commercial product (Zentrip®) was developed for people who suffered from motion sickness. In order to improve the mechanical strength of Zentrip®, OTF containing meclizine hydrochloride (MH) was designed and prepared using the solvent casting method. The characteristics of the prepared OTF were evaluated using micrometer, auto stripping tester, DSC, X-ray diffraction. ATR-FTIR was employed to investigate the interaction between drug and polymer. The thickness of MH OTF obtained was 0.116±0.004mm, the tensile strength was 17.37±1.54Nmm(-2) and the drug dissolution at 5min was more than 80% both in distilled water and 0.1mol/L HCL. DSC and XRD showed MH was amorphous in the polymer. ATR-FTIR indicated the MH molecules inserted into the network structure of polymer, which resulted in an inhibition of drug recrystallization. The Cmax of Zentrip® and MH OTF were 1.46±0.44μg/mL and 1.91±0.51μg/mL, and the AUC were 10.38±2.93μgh/mL and 13.74±3.23μgh/mL, respectively. Compared with Zentrip®, MH OTF successfully overcome the weakness of mechanical strength, possessed faster dissolution profile and showed bioequivalence in pharmacokinetics, deserving to a further development.


A cross-sectional study assessing the relationship between BMI, asthma, atopy, and eNO among schoolchildren.
Increased body weight may influence airway inflammatory mechanisms. To assess whether overweight-obesity (OW-O), evaluated as increased body mass index, is associated either with exhaled nitric oxide (eNO), a marker of airway inflammation, or with allergic sensitization in a large sample of children and adolescents. A cross-sectional, epidemiological study was performed on a population sample of schoolchildren evaluating 708 subjects (age 10-16 years; BMI 13-39 kg/m(2)) by respiratory health questionnaire, skin prick tests, spirometry, and eNO measure. Prevalence rates were: OW-O 16.4%, asthma ever (A) 11.9%, and rhinoconjunctivitis (RC) 14.8%. Asthma ever and allergic sensitization were significantly more frequent among OW-O (21.0 and 51.6%) than in non-OW-O (10.2 and 37.0%, respectively). The forced expiratory volume in 1 second (FEV(1))/forced vital capacity (FVC) ratio was not significantly different between OW-O and non-OW-O. Exhaled NO (median and interquartile range) was 15.3 (11.2-23.1) ppb in the overall sample, 20.3 (12.9-35.8) ppb among allergic subjects, and 13.9 (10.6-18.3) ppb among nonallergic subjects (P<.0001). No significant difference between OW-O and non OW-O subjects was found in eNO levels. Similarly, OW-O subjects with A or RC did not show significantly higher eNO levels than non-OW-O. In a logistic regression model, presence of allergic sensitization, A, and RC, and not OW-O, were significant predictors of increased eNO. In children, OW-O was not associated with increased eNO levels, but it was an independent risk factor for asthma and allergic sensitization.


Nationwide patterns of hydroxychloroquine dosing and monitoring of retinal toxicity in patients with systemic lupus erythematosus.
This study identified trends in hydroxychloroquine (HCQ) prescription and retinopathy screening in patients with systemic lupus erythematosus (SLE) according to clinical practice guidelines to minimise the risk of HCQ retinopathy. We used data from patients diagnosed with SLE between 2004 and 2019 from the National Health Insurance Service in Korea. To assess trends of daily dose per actual body weight (ABW), we performed an interrupted time-series analysis and identified effects after revision of guidelines. Among 38,973 patients with SLE, 28,415 (72.9%) were prescribed HCQ from 2004 to 2019. The proportion of patients using HCQ among SLE patients was 63% in 2004 and increased to 76% in 2019. The median daily dose per ABW for HCQ users decreased from 5.88 mg/kg in 2004 to 3.98 mg/kg in 2019, and from 5.45 mg/kg in 2005 to 4.17 mg/kg in 2019 for HCQ new users. The annual implementation rate of screening tests among HCQ new users increased from 3.5% in 2006 to 22.5% in 2019. Study results indicated that HCQ dosing management was adequate based on the revised guidelines. Although the implementation rate of retinal screening has increased, it is necessary to enhance awareness of retinal screening in clinical settings.


Effects of the Dietary Approaches to Stop Hypertension (DASH) eating plan on cardiovascular risks among type 2 diabetic patients: a randomized crossover clinical trial.
To determine the effects of the Dietary Approaches to Stop Hypertension (DASH) eating pattern on cardiometabolic risks in type 2 diabetic patients. A randomized crossover clinical trial was undertaken in 31 type 2 diabetic patients. For 8 weeks, participants were randomly assigned to a control diet or the DASH eating pattern. After following the DASH eating pattern, body weight (P = 0.007) and waist circumference (P = 0.002) reduced significantly. Fasting blood glucose levels and A1C decreased after adoption of the DASH diet (-29.4 ± 6.3 mg/dl; P = 0.04 and -1.7 ± 0.1%; P = 0.04, respectively). After the DASH diet, the mean change for HDL cholesterol levels was higher (4.3 ± 0.9 mg/dl; P = 0.001) and LDL cholesterol was reduced (-17.2 ± 3.5 mg/dl; P = 0.02). Additionally, DASH had beneficial effects on systolic (-13.6 ± 3.5 vs. -3.1 ± 2.7 mmHg; P = 0.02) and diastolic blood pressure (-9.5 ± 2.6 vs. -0.7 ± 3.3 mmHg; P = 0.04). Among diabetic patients, the DASH diet had beneficial effects on cardiometabolic risks.


The mechanism of free base formation from DNA by bleomycin. A proposal based on site specific tritium release from Poly(dA.dU).
Poly(dA.dU), which is specifically tritiated at the 1'-, 2'- (ribo configuration), 3'-, or 4'-position of deoxyuridine, has been synthesized and the fate of the tritium has been determined upon degradation of the polymer by bleomycin, Fe(II), and O2. No tritium is labilized from the 1'-3H-labeled polymer as 3H2O; however, the resulting 3-(uridin-1'-yl)-2-propenal (uracil propenal) has the expected specific activity. The 2'-3H-labeled polymer affords 3H2O and no label in the uracil propenal. This result and the lack of solvent incorporation into the uracil propenal suggest that proton abstraction from C-2' to afford the trans-propenal is highly stereospecific. For the 3'-3H-labeled polymer, 3H2O is formed and the specific activity of the uracil propenal is identical to that of the deoxyuridine. This suggests that the labilization of the 3'-H is exclusively associated with free uracil formation. 3H2O is also formed from the 4'-3H-labeled polymer. These findings along with previous studies are consistent with the formation of uracil propenal and free uracil by the trapping of the initially formed 4'-radical species by O2 or by a monooxygen species, respectively.


Role of 18F-FDG PET/CT in diagnosing peritoneal carcinomatosis in the restaging of patient with ovarian cancer as compared to contrast enhanced CT and tumor marker Ca-125.
To investigate the role of whole-body fluorine-18-2-deoxy-2-fluoro-d-glucose positron emission tomography/computed tomography ((18)F-FDG PET/CT) in the identification of peritoneal carcinomatosis in patients with ovarian cancer (OC). Seventy-nine patients with histologically proven stages III-IV OC who underwent (18)F-FDG PET/CT were studied retrospectively. We considered group A as 51 patients who also underwent computed-tomography with contrast-enhancement (CECT), and group B as 35 patients who had also been tested for biomarker Ca-125. Sensitivity, specificity, accuracy, positive predictive values (PPV) and negative predictive values (NPV) of (18)F-FDG PET/CT as compared to CECT and to Ca-125 were evaluated. (18)F-FDG PET/CT' sensitivity, specificity, accuracy, PPV and NPV for all 79 patients were: 85%, 92.31%, 88.61%, 91.89% and 85.71%, respectively. (18)F-FDG PET/CT sensitivity in group A was 78.6%, while it was 53.6% for CECT. (18)F-FDG PET/CT specificity, calculated in the same group, was 91.3%, while that of CECT was 60.9% (statistically significant difference, McNemar 4, P=0.039). Accuracy was 84.3% and 56.9%, respectively. (18)F-FDG PET/CT' sensitivity in group B was 86.4%, while that of Ca-125 was 81.8% (no statistical difference, McNemar 0, P=1). (18)F-FDG PET/CT specificity in group B was 84.6% while that of Ca-125 was 38.5% (clear but not statistically significant difference, McNemar 3.12, P=0.070). Accuracy calculated in the same group was 85.7% for (18)F-FDG PET/CT and 65.7% for Ca-125. (18)F-FDG PET/CT is a useful diagnostic tool when peritoneal biopsy cannot be performed and it can better select those who are candidates for adjuvant chemotherapy.


Rubber solvent: a clastogenic agent that fails to induce sister-chromatid exchanges.
Rubber solvent was tested for its ability to induce chromosome aberrations and sister-chromatid exchanges in human whole blood cultures. Following exposure to relatively low rubber solvent concentrations (0.0125% and greater) significant increases in the frequencies of chromatid gaps and breaks were observed. At higher rubber-solvent concentrations (0.05% and greater) there were also significant increases in the frequency of chromosome breaks. In contrast to the increase in chromosome aberrations following rubber-solvent exposure, rubber-solvent concentrations up to the toxic level failed to produce increases in the sister-chromatid exchange frequency.


Repeated haloperidol administration changes basal release of striatal dopamine and subsequent response to haloperidol challenge.
The effects of acute or repeated administration of haloperidol on release of dopamine (DA) and homovanillic acid (HVA) from striata of awake rats were studied using a microdialysis probe. A single injection of haloperidol (1 mg/kg, i.p.) produced a time-dependent increase in DA and HVA in the perfusate. Comparative studies in rats anesthetized with 300 mg/kg of chloral hydrate given i.p. found that anesthesia decreased the basal release of DA, but not HVA, and significantly blocked haloperidol-induced increases in DA, while haloperidol-induced increases in HVA were not affected. Studies done in awake rats found that 21 repeated daily injections of haloperidol increased the basal release of DA, but not HVA. Subsequent challenge with haloperidol indicated a significant decrease in responsiveness to haloperidol-induced release of DA, but not HVA, in chronically dosed rats. These data suggest that repeated exposure to haloperidol causes a compensatory increase in extracellular DA release. That these compensatory changes may be associated with the increased therapeutic efficacy or extrapyramidal side effects of neuroleptics following repeated dosing warrants further study.


Molecular mechanisms of microRNAs in glioblastoma pathogenesis.
Glioblastoma (GBM) is human's most prevalent and severe brain cancer. Epigenetic regulators, micro(mi)RNAs, significantly impact cellular health and disease because of their wide range of targets and functions. The "epigenetic symphony" in which miRNAs perform is responsible for orchestrating the transcription of genetic information. The discovery of regulatory miRNA activities in GBM biology has shown that various miRNAs play a vital role in disease onset and development. Here, we summarize our current understanding of the current state-of-the-art and latest findings regarding the interactions between miRNAs and molecular mechanisms commonly associated with GBM pathogenesis. Moreover, by literature review and reconstruction of the GBM gene regulatory network, we uncovered the connection between miRNAs and critical signaling pathways such as cell proliferation, invasion, and cell death, which provides promising hints for identifying potential therapeutic targets for the treatment of GBM. In addition, the role of miRNAs in GBM patient survival was investigated. The present review, which contains new analyses of the previous literature, may lead to new avenues to explore in the future for the development of multitargeted miRNA-based therapies for GBM.


A reproducible procedure for primary culture and subsequent maintenance of multiple lines of human skin fibroblasts.
Cultured fibroblasts are a valuable tool to study many cellular processes and their modification by aging. Fibroblasts are a useful cell type in which to study many diseases, including those of the nervous system, in which a strong genetic component is suspected. Fibroblasts permit the study of multiple, dynamic processes in living cells, while avoiding the effect of the dying process and post-mortem artifacts that limit other approaches. For results to be comparable across time in one laboratory or consistent between laboratories, the detailed culture techniques require meticulous care and replicability. Lack of attention to detail in initial stages can lead to selection of different cell populations. Small variations in othe variables such as batches of serum can significantly alter growth rates and comparisons of cells from controls and Alzheimer patients. The aim of this paper is to present a detailed protocol for comparison of multiple cell lines from many patients. An example of using this approach to study growth and phase out (i.e., senescence) of cells from Alzheimer patients is presented. This procedure represents a modification of an earlier protocol (Cristofalo and Charpentier, 1980).


Total pancreatectomy in six patients with intraductal papillary mucinous tumour of the pancreas: the treatment of choice.
Intraductal papillary mucinous tumours (IPMT) were described as a distinct entity in 1982.The extent of surgical resection remains controversial. Six patients with a diffuse dilatation of the main pancreatic duct were treated with total pancreatectomy for cure of IPMT. Histological examination showed one IPM adenoma, four IPM non-invasive carcinomas and one IPM invasive carcinoma. In all but one case multifocal extensive intraductal changes were found, affecting either most of the pancreas or the whole organ. All patients survived the operation and remain alive 5-56 months later. Post-pancreatectomy diabetes has been moderately well controlled. IPMTs represent a subgroup of pancreatic neoplasms with a favourable prognosis, and the resection should aim at removing all dysplastic foci. In cases with diffuse dilatation of the main pancreatic duct, widespread tumour involvement of the duct system can be expected, so total pancreatectomy should be the operation of choice.


The need for prophylactic treatment to the central nervous system in patients with aggressive non-Hodgkin's lymphoma.
During 2 years, 6 out of 34 patients presenting with newly diagnosed non-Hodgkin's lymphoma developed central nervous system (CNS) complications. All were male and had diffuse, undifferentiated lymphomas. In all 6 patients, CNS disease developed during, or shortly after, treatment. We suggest that a group at high risk from these complications can be identified and should be considered for a trial of prophylaxis.


In silico analysis and in vitro evaluation of immunogenic and immunomodulatory properties of promiscuous peptides derived from Leishmania infantum eukaryotic initiation factor.
It is generally considered as imperative the ability to control leishmaniasis through the development of a protective vaccine capable of inducing long-lasting and protective cell-mediated immune responses. In this current study, we demonstrated potential epitopes that bind to H2 MHC class I and II molecules by conducting the in silico analysis of Leishmania infantum eukaryotic Initiation Factor (LieIF) protein, using online available algorithms. Moreover, we synthesized five peptides (16-18 amino acids long) which are part of the N-terminal portion of LieIF and contain promising MHC class I and II-restricted epitopes and afterwards, their predicted immunogenicity was evaluated in vitro by monitoring peptide-specific T-cell responses. Additionally, the immunomodulatory properties of these peptides were investigated in vitro by exploring their potential of inducing phenotypic maturation and functional differentiation of murine Bone-Marrow derived Dendritic Cells (BM-DCs). It was revealed by our data that all the synthetic peptides predicted for H2 alleles; present the property of immunogenicity. Among the synthetic peptides which contained T-cell epitopes, the peptide 52-68 aa (LieIF_2) exhibited immunomodulatory properties with the larger potential. LieIF_2-pulsed BM-DCs up-regulated the expression of the co-stimulatory surface molecules CD80 and CD86, as well as the production of the proinflammatory cytokine TNF-α and of the Th1-polarizing cytokines IL-12 and IFN-γ. The aforementioned data suggest that selected parts of LieIF could be used to develop innovative subunit protective vaccines able to induce effective immunity mediated by MHC class I-restricted as well as class II-restricted T-cell responses.


Oxford foot model kinematics in landings: A comparison between professional dancers and non-dancers.
Dancers frequently perform jump-landing activities, with the foot-ankle complex playing an essential role to attenuate the landing forces. However, scarce research has been conducted in professional dancers multi-segmented foot in landings. The aim of this study was to compare the multi-segmented foot kinematics between professional dancers and non-dancers, during forward and lateral single-leg jump-landings. Descriptive group comparison. Marker trajectories and synchronized ground reaction forces of 15 professional dancers and 15 non-dancers were collected using motion capture and a force plate, during multidirectional single-leg jump-landings. Sagittal and frontal hindfoot-tibia, forefoot-hindfoot, and hallux-forefoot kinematics of the multi-segmented foot model were computed at initial contact, peak vertical ground reaction force and peak knee flexion. Repeated measures ANOVAs were conducted (p < 0.05). Professional dancers landed with higher hindfoot-tibia and forefoot-hindfoot plantarflexion angles at initial contact (p < 0.001), and hindfoot-tibia dorsiflexion angles at peak vertical ground reaction force and peak knee flexion (p < 0.001) than non-dancers. Also, dancers exhibited higher sagittal hindfoot-tibia and forefoot-hindfoot excursions than non-dancers (p < 0.001). No statistically significant differences were found in the frontal plane. The multi-segmented foot allows a comprehensive kinematic analysis of the different foot joints. In jump-landings, professional dancers higher hindfoot-tibia, and forefoot-hindfoot plantarflexion at initial contact, compared to non-dancers, contributed to a subsequent higher foot joints excursion. This pattern is commonly linked to a better shock absorption mechanism in landings.


Outcomes of pelvic exenteration for locally advanced primary rectal cancer: Overall survival and quality of life.
Radical surgery with pelvic exenteration offers the only potential for cure in patients with locally advanced primary rectal cancer. This study describes the clinical and patient-reported quality of life outcomes over 12 months for patients having pelvic exenteration for locally advanced primary rectal cancer at a specialised centre for pelvic exenteration. Clinical data of consecutive patients undergoing pelvic exenteration for locally advanced primary rectal cancer and patient-reported outcomes were collected at baseline, hospital discharge and at 1, 3, 6, 9 and 12 months. Patient-reported outcomes included cancer-specific quality of life (QoL) and physical and mental health status. Quality of life trajectories were modelled over the 12 months from the date of surgery using linear mixed models. 104 patients with locally advanced rectal cancer underwent pelvic exenteration at Royal Prince Alfred Hospital, Sydney, between December 1994 and October 2014. Complete soft tissue exenteration was performed in 38%. A clear margin was obtained in 86% with a 62% overall five-year survival. QoL outcome questionnaires were completed by 62% of patient cohort. The average FACT-C score returned to pre-surgery QoL by 2 months after surgery, and the average QoL continued to increase slowly over the 12 months. Our results support an aggressive approach to advanced primary rectal cancer and lend weight to the oncological role of pelvic exenteration for this group of patients. Quality of life improves rapidly after pelvic exenteration for locally advanced primary rectal cancer and continues to improve over the first year.


Dihydrochalcone Compounds Isolated from Crabapple Leaves Showed Anticancer Effects on Human Cancer Cell Lines.
Seven dihydrochalcone compounds were isolated from the leaves of Malus crabapples, cv. "Radiant", and their chemical structures were elucidated by UV, IR, ESI-MS, ¹H-NMR and (13)C-NMR analyses. These compounds, which include trilobatin (A1), phloretin (A2), 3-hydroxyphloretin (A3), phloretin rutinoside (A4), phlorizin (A5), 6''-O-coumaroyl-4'-O-glucopyranosylphloretin (A6), and 3'''-methoxy-6''-O-feruloy-4'-O-glucopyranosyl-phloretin (A7), all belong to the phloretin class and its derivatives. Compounds A6 and A7 are two new rare dihydrochalcone compounds. The results of a MTT cancer cell growth inhibition assay demonstrated that phloretin and these derivatives showed significant positive anticancer activities against several human cancer cell lines, including the A549 human lung cancer cell line, Bel 7402 liver cancer cell line, HepG2 human ileocecal cancer cell line, and HT-29 human colon cancer cell line. A7 had significant effects on all cancer cell lines, suggesting potential applications for phloretin and its derivatives. Adding a methoxyl group to phloretin dramatically increases phloretin's anticancer activity.


Ototoxicity of olive oil in a chinchilla animal model.
Olive oil is often used by patients to soften ear wax or to relieve ear canal obstruction. It is also sold in drugstores as a cerumenolytic. To date, no study has assessed the safety of ototopical olive oil on hearing in the presence of tympanic membrane perforation. The present study aimed to assess the safety of ototopic olive oil on hearing in the presence of tympanic membrane perforation. Prospective, randomized, controlled trial in a chinchilla animal model. Eleven chinchillas underwent bilateral myringotomy. In each animal, one ear was randomly assigned to receive olive oil (experimental ear), while the contralateral control ear received normal saline. Auditory brain response (ABR) test was performed at baseline and then 7, 14, and 30 days following the application. At 30 days follow-up, there was no significant change in auditory brain response thresholds at 8, 16, 20, or 25 kHz. Scanning electron microscope imaging showed no damage to the hair cells. Olive oil does not seem to cause hearing loss in chinchillas with perforated tympanic membranes. Future clinical studies are required.


Morphology regulation in vascular endothelial cells.
Morphological change in endothelial cells is an initial and crucial step in the process of establishing a functional vascular network. Following or associated with differentiation and proliferation, endothelial cells elongate and assemble into linear cord-like vessels, subsequently forming a perfusable vascular tube. In vivo and in vitro studies have begun to outline the underlying genetic and signaling mechanisms behind endothelial cell morphology regulation. This review focuses on the transcription factors and signaling pathways regulating endothelial cell behavior, involved in morphology, during vascular development.


Amphibian and reptile phenology: the end of the warming hiatus and the influence of the NAO in the North Mediterranean.
In the south of France, the so-called climate hiatus from 1998 to 2013 was associated with a late winter cooling which has affected the phenology of several reptiles and amphibian species, delaying their dates of first appearances in spring. This episode has been related to a period of frequently negative values of the North Atlantic Oscillation index (NAOi). The recent increase of this index after this episode marks the end of the "hiatus" and provides an opportunity to verify the impact of the North Atlantic Oscillation (NAO) on the fauna of the North Mediterranean region. Most of the emergence dates of amphibians and reptiles in spring have rapidly advanced from 1983 to 1997 and then receded or stabilized from 1998 to 2010. They began to advance again since 2010. These phenological changes covary with the temperature of February-March in the study area, which is itself related to the variations of the NAO index. These changes confirm the influence of the NAO on the phenology of terrestrial organisms in northern Mediterranean where its influence is sometimes assumed to be attenuated.


4. Steroid pathophysiology of BPH. Correlative morphological and biochemical investigations on the stromal tissue of the human prostate.
Light and electron microscopic measurements show that tissue overgrowth in human benign prostatic hyperplasia is mostly due to an increase in stromal tissue. There is evidence that interaction between stromal and epithelial cells play a critical role in the regulation of the coordinated growth of the prostatic gland. Stromal response to hormones is virtually not completely understood, however at present there are strong indications that it is the androgens above all which control the function of the stromal part of the human prostate. It has been shown that the endogenous concentration of 17 beta-estradiol in the stromal compartment of the guinea pig prostate exceeded that of the blood plasma and prostatic epithelium. In an attempt to shed light on the action of endogeneous androgens and estrogens on the stromal tissue of the human prostate, selective androgen and estrogen antagonists as well as bromocriptine were used to study the stromal reaction pattern in human prostatic hyperplasia.


What are the factors affecting older adults' experience of unmet healthcare needs amid the COVID-19 pandemic in Korea?
Unmet healthcare need is a critical indicator, showing a plausible picture of how the healthcare system works in the unprecedented pandemic situation. It is important to understand what factors affect healthcare services of older adults in the midst of the outbreak, as this could help identify service- and performance-related challenges and barriers to the healthcare system. This study aimed to identify factors associated with unmet healthcare needs among the older Korean population amid the COVID-19 pandemic. Cross-sectional data were used from the Experience Survey on Healthcare Use of Older Adults during the COVID-19 (COVID-19 Survey) in Korea (n = 1,917). Our main outcome, unmet healthcare need, was measured based on self-reported experience of overall, regular, and irregular outpatient care services-related unmet healthcare needs. Independent variables were selected based on previous studies on determinants of unmet healthcare need during the COVID-19 pandemic and Andersen's expanded behavioural model, which theorizes that healthcare-seeking behaviours are driven by psychosocial, enabling, and need factors. Using multiple logistic regression models, we identified a good understanding of the nation's health system was associated with lower likelihood of all types of unmet healthcare needs among older Korean adults (OR: 0.39, 95%CI: 0.25-0.61; OR: 0.36, 95%CI: 0.20-0.63; OR: 0.41, 95%CI: 0.23-0.75). Decreased social activities (i.e., shopping and visiting family members) and worsened psychological health issues (i.e., increased anxiety & nervousness and greater difficulty sleeping) were also factors affecting overall and irregular outpatient services-related unmet needs. To ensure timely access to necessary healthcare services for older adults in the era of the COVID-19 outbreak, improving older adult's understanding on how the healthcare system works is necessary. Moreover, changes in psychological condition and daily activities due to COVID-19 should be considered as possible barriers to healthcare services among older adults during the global pandemic.


Regulation of aromatase by nuclear receptors.
We investigated the effects of a nuclear receptor system constituted by retinoid X receptor (RXR) and its heterodimer partner on the aromatase activity in a cultured MCF-7 human breast cancer cell line and also in human ovarian granulosa cells, using each selective ligand for retinoic acid receptor, RAR (TTNPB), retinoid X receptor, RXR (LG100268), PPARgamma (troglitazone), and vitamin D3 receptor (cholecalciferol). In MCF-7 cells, the combined treatment with TTNPB and LG100268 caused a dramatic stimulation of the aromatase activity. The combined treatment with other ligand and LG100268 had little or no effect on the aromatase activity. The increase in the aromatase activity by TTNPB plus LG100268 was accompanied by an increase in the P450arom mRNA levels, which was also found to be related to the specific usage of promoter 1a of the CYP19 gene. These results suggest that a nuclear receptor system constituted by a RAR:RXR heterodimer is involved in the regulation of aromatase activity in MCF-7 breast cancer cells. In cultured human ovarian granulosa cells obtained from patients who underwent in vitro fertilization, troglitazone or LG100268 alone decreased the aromatase activity, while the combined treatment caused an even greater reduction in this activity. Little effect of other specific ligands for RXR heterodimer partners may support the notion that the effects of troglitazone and/or LG100268 in human granulosa cells may be mediated through the specific activation of PPARgamma:RXR heterodimer system. Since similar manners of effects of several PPARgamma ligands and/or LG100268 on the aromatase activity were observed in a newly established human ovarian granulosa cancer cell line, KGN, we performed the detailed analysis of the mechanisms of these effects using this cell line. As a result, the inhibitory effect of aromatase activity by troglitazone plus LG100268 was accompanied by the decrease of P450arom mRNA level. Furthermore, the loss of P450arom expression was considered to be due to both the decreased transcription and rapid degradation of its RNA based on the studies of nuclear run-on assay and RNA stability assay. In conclusion, RAR:RXR and PPARgamma:RXR heterodimer nuclear receptor systems may be other important modulators of estrogen production in human breast cancer cells and ovarian granulosa cells, respectively.


Late psychosocial sequelae in Hodgkin's disease survivors: a French population-based case-control study.
To evaluate late psychosocial sequelae in long-term survivors of Hodgkin's disease (HD) in the population of Calvados, France. Ninety-three patients issued from the Calvados General Tumor Registry, treated from 1978 to 1990, free of relapse and second malignancy since January 1991, were enrolled onto cross-sectional case-control study. One hundred eighty-six healthy controls, matched for sex, age, and residency, were selected at random from electoral rolls. Two self-administered questionnaires were mailed in the spring of 1995. Compared with controls, HD patients reported (1) more physical (P < .001), role (P < .001), and cognitive (P = .015) functioning impairments, as well as dyspnea (P < .001) and chronic fatigue (P = .025), while no statistical difference was found in global health status; (2) to be more often childless (P = .04), fewer divorces or separations (P = .013), fewer changes in relationships with friends (P = .012), similar proportions at work but less ambitious professional plans (P < .001), and greater difficulties in borrowing from banks (P < .001); (3) a slight increase in the number of visits to a general practitioner (P = .05) and greater consumption of medical resources (mainly thyroid extracts, P = .05). The study demonstrated that French long-term HD survivors have good global health status and good psychologic, familial, and professional status, although difficulties in borrowing from banks remain a major limitation in daily life. Although physical, role, and cognitive functioning impairments persist that might limit their activities, HD survivors seem to have learned to cope with problems related to their disease and its treatment.


Intra-individual variability in the response to anodal transcranial direct current stimulation.
To test the intra-individual reliability in response to anodal transcranial direct current stimulation (AtDCS). 45 healthy subjects received AtDCS (1 mA, 13 min) in two separate sessions, 6-12 months apart. Motor evoked potentials were collected at baseline and then at 5-min intervals after AtDCS for 1h. Short intracortical inhibition (SICI) was assessed at minutes 6 and 46 after AtDCS. AtDCS increased cortical excitability over minutes 0-30 post-stimulation in both sessions, with fair intra-individual reliability. 60% and 64% of subjects responded with the expected increase in cortical excitability in each session, respectively. 69% of the subjects maintained their response pattern between sessions during this timeframe. However, there were no significant effects on cortical excitability over the full hour post AtDCS in either session. SICI showed fair intra-individual reliability 6 min after AtDCS. A change in cortical excitability in the first half-hour post-AtDCS may be a good predictor of the response in a subsequent session. Furthermore, minute 15 post-stimulation showed the maximum increase in cortical excitability in both sessions. We show for the first time that intra-individual variability is lower than inter-individual variability, and with fair intra-individual inter-sessional reliability for 30 min after AtDCS-subjects are likely to maintain their response patterns to tDCS between sessions, with implications for experimental and therapeutic applications of tDCS.


Role of heat shock proteins in protection from and pathogenesis of infectious diseases.
Increased synthesis of heat shock proteins (hsp) occurs in prokaryotic and eukaryotic cells when they are exposed to stress. By increasing their hsp content, cells protect themselves from lethal assaults, primarily because hsp interfere with the uncontrolled protein unfolding that occurs under stress. However, hsp are not produced only by stressed cells; some hsp are synthesized constitutively and perform important housekeeping functions. Accordingly, hsp are involved in the assembly of molecules which play important roles in the immune system. It is not surprising that due to their wide distribution and their homology among different species, hsp represent target antigens of the immune response. Frequent confrontation of the immune system with conserved regions of hsp which are shared by various microbial pathogens can potentiate antimicrobial immunity. However, long-term confrontation of the immune system with hsp antigens which are similar in the host and invaders may convert the immune response against these host antigens and promote autoimmune disease. This review provides an overview of the role of hsp in immunity with a focus on infectious and autoimmune diseases.


Optogenetics in the cerebellum: Purkinje cell-specific approaches for understanding local cerebellar functions.
The cerebellum consists of the cerebellar cortex and the cerebellar nuclei. Although the basic neuronal circuitry of the cerebellar cortex is uniform everywhere, anatomical data demonstrate that the input and output relationships of the cortex are spatially segregated between different cortical areas, which suggests that there are functional distinctions between these different areas. Perturbation of cerebellar cortical functions in a spatially restricted fashion is thus essential for investigating the distinctions among different cortical areas. In the cerebellar cortex, Purkinje cells are the sole output neurons that send information to downstream cerebellar and vestibular nuclei. Therefore, selective manipulation of Purkinje cell activities, without disturbing other neuronal types and passing fibers within the cortex, is a direct approach to spatially restrict the effects of perturbations. Although this type of approach has for many years been technically difficult, recent advances in optogenetics now enable selective activation or inhibition of Purkinje cell activities, with high temporal resolution. Here we discuss the effectiveness of using Purkinje cell-specific optogenetic approaches to elucidate the functions of local cerebellar cortex regions. We also discuss what improvements to current methods are necessary for future investigations of cerebellar functions to provide further advances.


Mutation in pfmdr1 gene in chloroquine-resistant Plasmodium falciparum isolates, Southeast Iran.
The main objective of the present study was to detect point mutations at positions 86, 184, 1034, 1042, and 1246 of the Plasmodium falciparum multidrug resistance gene (pfmdr1) in blood samples collected from malaria patients in Chabahar, a harbor city located in Southeast Iran. Twenty-six blood samples from patients infected with P. falciparum, who had a chloroquine (CQ) response failure, were collected pre-treatment. Following treatment with CQ, drug susceptibility was assessed using an in vivo test. Molecular detection of single nucleotide polymorphisms (SNPs) was carried out using the LightCycler hybridization probe assay. The pfmdr1 N86Y mutation was found in six isolates (23.1%). Mutations at the four other positions were not observed in any isolates. The present study showed no mutation at codon positions 184, 1034, 1042, and 1246 of pfmdr1 in any of the Iranian P. falciparum isolates; thus these alleles cannot serve as markers for CQ resistance in Iran.


Cerebral microvessels and derived cells in tissue culture: isolation and preliminary characterization.
Microvessels isolated from mouse forebrain were used as the source material for the derivation of cerebral vascular endothelium and smooth-muscle cells in culture. The microvessels were isolated by a mechanical dispersion and filtration technique, and were maintained in vitro as organoid cultures. A microvessel classification system was developed and proved to be useful as a tool in monitoring culture progress and in predicting the type(s) of microvessel(s) that would give rise to migrating and/or proliferating cells. The isolated cerebral microvessels were heterogeneous in diameter, size of individual vascular isolate, and proliferative potential. The isolated microvessels ranged in diameter from 4 micron to 25 micron and in size from a single microvascular segment to a large multibranched plexus with mural cells. The initial viability, determined by erythrosin B exclusion, was approximately 50% on a per cell basis. All microvessel classes had proliferative potential although the rate and extent of proliferation were both microvessel class- and density-dependent. The smaller microvessels gave rise to endothelial cells, whereas the large microvessels gave rise to endothelial and smooth-muscle cells. The viability and progress of a microvessel toward derived cell proliferation seemed to be directly proportional to the number of mural cells present.


Amphiphilic cationic polymers as effective substances improving the safety of use of body wash gels.
Cationic surfactants have a wide range of applications in the cosmetic industry. The present study is an attempt to determine the effect of an amphiphilic cationic polymer - a cationic derivative of guar gum - on the safety of use of body wash cosmetics. Model body wash gels based on an anionic surfactant, containing 0.05, 0.25 and 0.5% of the studied compound, were subjected to analyses in order to evaluate their skin irritation effect and the ability to solubilize model sebum. Cell culture studies were also carried out. In addition, the effect of the cationized guar gum derivative on the key quality parameters of body wash cosmetics (viscosity and foaming properties) was assessed. The analyses showed that the addition of the cationic guar gum derivative induced a significant increase in the safety of use of the model body wash gels without causing any impairment of the functional properties of the product.


Real time metagenomics: using k-mers to annotate metagenomes.
Annotation of metagenomes involves comparing the individual sequence reads with a database of known sequences and assigning a unique function to each read. This is a time-consuming task that is computationally intensive (though not computationally complex). Here we present a novel approach to annotate metagenomes using unique k-mer oligopeptide sequences from 7 to 12 amino acids long. We demonstrate that k-mer-based annotations are faster and approach the sensitivity and precision of blastx-based annotations without loosing accuracy. A last-common ancestor approach was also developed to describe the members of the community.


Needle tract implantation of hepatoblastoma after percutaneous needle biopsy: report of a case.
A 13-year-old boy was referred to us for investigation of a giant liver mass, approximately 16 cm in diameter. Sonographically guided percutaneous needle biopsy was performed and histological examination revealed a fetal-type hepatoblastoma. After four courses of chemotherapy, we performed a left hepatic trisegmentectomy. Follow-up computed tomography, 55 months after the surgery, showed a 1-cm tumor on the route of the preoperative needle biopsy. A second laparotomy revealed a peritonealised tumor, which was excised. The histology of this tumor was identical to that of the primary hepatoblastoma. To our knowledge, this is only the second report of needle tract implantation of hepatoblastoma after percutaneous needle biopsy.


Altered vancomycin dose vs. serum concentration relationship in burn patients.
Drug elimination in patients sustaining serious thermal injury may be altered, resulting in an increased clearance and shortened half-life. Nine burn and eight medical/surgical patients with normal renal function were studied prospectively. Doses were adjusted to achieve peak and trough vancomycin serum concentrations within a narrow range. No significant difference between the groups was noted in terms of demographic characteristics, creatinine clearance, or vancomycin serum concentrations. However, the difference in daily dose needed to maintain the specified serum concentrations was significantly greater for burn patients (p less than 0.02). Burn patients also had to be dosed significantly more often than medical/surgical patients to achieve peak and trough vancomycin serum concentrations within the desired range (p less than 0.02). The elimination half-life in burn patients was significantly shorter than that in control patients (p less than 0.001). Because of the unusually high dosage requirements in burn patients, along with their poor predictability, individualization of therapy with vancomycin serum concentrations is recommended to ensure a successful therapeutic outcome.


Rodent-associated Bartonella febrile illness, Southwestern United States.
Serum specimens from 114 patients hospitalized with a febrile illness were tested with an indirect immunofluorescence assay (IFA) using Bartonella antigens prepared from 6 species of sigmodontine rodents and 3 known human Bartonella pathogens: B. henselae, B. quintana, and B. elizabethae. Acute- and convalescent-phase serum samples from 5 of these patients showed seroconversion with an IFA titer >512 to rodent-associated Bartonella antigens. The highest titer was against antigen derived from the white-throated woodrat (Neotoma albigula), although this rodent is not necessarily implicated as the source of infection. Three of the 5 who seroconverted showed no cross-reaction to the 3 Bartonella human pathogens. Common clinical characteristics were fever, chills, myalgias, leukopenia, thrombocytopenia, and transaminasemia. Although antibodies to Bartonella are cross-reactive, high-titer seroconversions to rodent-associated Bartonella antigens in adults with common clinical characteristics should stimulate the search for additional Bartonella human pathogens.


Circadian biology: sleep-styles shaped by light-styles.
Light and darkness are the main time cues synchronising all biological clocks to the external environment. This little understood evolutionary phenomenon is called circadian entrainment. A new study illuminates our understanding of how modern light- and lifestyles compromise circadian entrainment and impact our biological clocks.


Nitrogen sparing effects and mechanisms of branched-chain amino acids in the injured rat.
A series of experiments in a rat injury model were designed to elucidate the role and mechanisms of branched-chain amino acids in the post-injury catabolism. Our results suggest that: 1. Nutritional support can maintain nitrogen equilibrium in the early post-operative state. 2. Branched chain amino acids exert a nitrogen sparing effect and thus prevent or minimise post-operative catabolism. 3. Increasing the amount of infused branched chain amino acids results in nitrogen retention. 4. A balanced amino-acid mixture containing 45 per cent branched chain amino acids seems to be optimal for nutritional support in the post-injury state.


Purification of exosome-enriched proteins produced in a Drosophila cell line by size exclusion chromatography.
Exosomes are a class of extracellular vesicles that play a role in intercellular signaling under diverse contexts. Here, we describe a protocol that has been optimized for the isolation and characterization of exosomes from a Drosophila melanogaster cell line using size exclusion chromatography (SEC). The specific focus of this protocol was to examine the starvation-induced exosome loading of a protein. For complete details on the use and execution of this protocol, please refer to Pandey et al. (2021).1.


Clinical diagnostic accuracy of basal cell carcinoma.
The clinical diagnostic accuracy and index of suspicion of basal cell carcinoma were calculated. The data were compiled from dermatology residents, full-time dermatology university faculty, and dermatologists in private practice.


Enhanced Oral Bioavailability of Curcumin Using a Supersaturatable Self-Microemulsifying System Incorporating a Hydrophilic Polymer; In Vitro and In Vivo Investigations.
A supersaturatable self-microemulsifying drug delivery system (S-SMEDDS) with a reduced amount of surfactant and incorporation of a polymer precipitation inhibitor, Eudragit® E PO was developed. The optimized S-SMEDDS formulation (SS-15) consisted of 55% surfactants, 40% oils, and 5% Eudragit® E PO (curcumin at 44.4 mg/g of the formulation). The precipitation profiles from the supersaturation assay revealed that the curcumin S-SMEDDS performed as a better inhibitor of curcumin precipitation in simulated gastric fluid over a 240-min study than the normal curcumin SMEDDS and an aqueous curcumin suspension. In addition, the mean droplet size of the curcumin S-SMEDDS (21.6 ± 0.1 nm) was significantly smaller than the SMEDDS (28.1 ± 0.3 nm). The curcumin S-SMEDDS exhibited a threefold reduction of Caco-2 cell toxicity when compared to the curcumin SMEDDS because of the reduced toxic effect of the surfactant present in the SMEDDS formulation. In addition, the absorptive permeability across the Caco-2 monolayer of curcumin in the S-SMEDDS was significantly higher than for the unformulated curcumin (~ 5-folds). The plasma concentration-time profiles from the oral absorption studies in rats dosed with the curcumin S-SMEDDS showed a 1.22- and 53.14-fold increased absorption of curcumin, compared to the SMEDDS and the aqueous suspension, respectively. The curcumin S-SMEDDS was stable under both intermediate and accelerated conditions after 6 months of storage.


Catechin inhibits Candida albicans dimorphism by disrupting Cek1 phosphorylation and cAMP synthesis.
Candida albicans is a fungal pathogen that undergoes dimorphism (transformation from a yeast form to a hyphal form), wherein, the yeast form is identified as a disseminating form that plays a critical role in the early stages of Candida disease progression, while the hyphal form is found to exert additional pathogenicity by adapting to various environmental conditions. Here, we elucidated the effects of catechin on C. albicans hyphal formation. Flow cytometry analysis showed catechin inhibited FCS-induced hyphal formation. Moreover, hypha-specific gene expression in MAP kinase cascade and cAMP pathway was decreased ascribable to catechin. Furthermore, through Western blotting and cAMP synthesis analysis, we found catechin obstructs Cek1 phosphorylation in MAP kinase cascade and suppresses cAMP synthesis. These results suggest that catechin possesses anti-dimorphism activity by interfering with in vitro signal transduction. Similarly, this highlights the possible application of catechin in clinical therapy for the management and prevention of candidosis.


Measurement of glycoprotein content and cell kinetics in preulcerous gastric mucosa.
In an attempt to evaluate the role of gastric mucosal defense factors in ulcerogenesis, we measured the levels of glycoproteins in the mucosa as well as mucosal cell turnover in the preulcerous phase and compared these parameters to the normal mucosa in the same animal. Ovalbumin-presensitized Praomys (Mastomys) natalensis were challenged in the gastric wall with ovalbumin and a gastric ulcer developed at the challenge site 3 days later as a result of a mucosal anaphylactic reaction. This model enabled us to study the events occurring at the site of a future ulceration. Gas-liquid chromatographic determination of mucosal glycoproteins showed that the normal and preulcerous mucosae had similar levels. Cell turnover, determined by [3H]thymidine incorporation, was stimulated as a result of the preulcerous anaphylactic reaction at 24 hours postchallenge whereas at 48 hours the values were not different from those obtained in controls. These results suggest that the pathogenesis of anaphylactic gastric ulcer involves a change in cell turnover but no changes in the production of gastric mucus.


A comprehensive update on Capsicum proteomics: Advances and future prospects.
Capsicum belonging to the family Solanaceae is one of the most widely consumed crops in the world as a vegetable, spice and a raw salad and is distinctly valuable for its spicy pungent flavour. Proteomic investigation of crop plants is an essential step towards deciphering the functional basis of traits in an organism and to deepen our understanding on the regulation of various developmental patterns, biotic, and abiotic stress response and tolerance mechanisms. The differential proteome expression profiling of tissues during different developmental stages and under different conditions may indicate the specific proteome dynamics involved in the developmental programs and under stress conditions. Although substantial progress in proteomics of other Solanaceae plants has been made in the past two decades, a comprehensive review on Capsicum proteomics is still lacking. This review provides updated information on the advancement of Capsicum proteomic study in cytoplasmic male sterility, during fruit development and ripening, and under different biotic and abiotic stresses. Although limited information is available on the post translational protein modifications in Capsicum, a brief outline is given at the end detailing various post translational modifications. This proteomic update on Capsicum will be useful for future studies aimed at Capsicum improvement programs.


Highly sensitive enumeration of circulating tumor cells in lung cancer patients using a size-based filtration microfluidic chip.
Circulating tumor cells (CTCs) in the peripheral blood could serve as a surrogate marker for the diagnosis of cancer metastasis and for therapeutic evaluation. However, the separation and characterization of CTCs is technically challenging owing to the extremely low number of CTCs present. Here we developed a size-based and high-throughput microfluidic chip, which exploits filtration microchannels to isolate the relatively larger CTCs from the rest of the blood constituents. High isolation efficiency of our microfluidic chip was demonstrated with three lung cancer cell lines spiked in blood samples at an optimal flow rate of 0.4 mL/h. The average recovery rates of 96%, 95% and 92% were obtained for A549, SK-MES-1, and H446, respectively. To clinically validate the chip, we also employed it to isolate CTCs from 59 lung cancer patients. CTCs were detected in 96.7% of patients with the mean number of 18.6 cells/mL, which was significantly higher than normal controls (P<0.05). The work here indicates that the size-based microfluidic platform with the advantage of capturing tumor cells without reliance on cell surface expression markers can provide a novel, inexpensive and effective tool for CTC detection and evaluation of cancer status.


Brassinosteroids induce plant tolerance against phenanthrene by enhancing degradation and detoxification in Solanum lycopersicum L.
Polycyclic aromatic hydrocarbons (PAHs) are toxic to both plants and animals. The enhancement of plant tolerance and detoxification capacity is important for the plant-based remediation of PAHs. Therefore, we investigated the effects of 24-epibrassinolide (EBR) on the metabolism of a three-ringed PAH (phenanthrene-PHE) and subsequent stress tolerance in tomato (Solanum lycopersicum L.) plants. Exposure to PHE (300 μM) for 21 d significantly decreased biomass and net CO(2) assimilation (P(n)) but induced photoinhibition, malondialdehyde (MDA), H(2)O(2) and antioxidant enzymes. Obvious ultrastructural alterations were observed in the PHE-treated root tip cells. Importantly, the foliar application of EBR (0.1 μM) significantly increased biomass, P(n) and antioxidant enzyme activities but decreased MDA and H(2)O(2) compared with PHE alone and saved the root cells from severe damage. The expression of detoxification genes (CYP90b3, GSH1, GST1), reduced glutathione (GSH) content and glutathione S-transferase activity in the EBR+PHE-treated plants were higher than those of PHE alone. Additionally, lower levels of PHE residues in the roots were observed as a result of EBR+PHE treatment. Taken together, our results strongly suggest an enhanced and coordinated detoxification and degradation of PHE by EBR.


Upper airway dimensions in patients undergoing orthognathic surgery: a systematic review and meta-analysis.
The objective of this systematic review was to evaluate the effect of different types of orthognathic surgery on the dimensions of the upper airways assessed using three-dimensional images. An electronic search was performed in Cochrane Library, Medline, Scopus, VHL, Web of Science, and the System for Information on Grey Literature in Europe, ending January 2015. Inclusion criteria encompassed clinical studies in humans, patient age >15 years, patients submitted to maxillary or mandibular advancement or setback surgery, isolated or in combination, and presentation of airway measures, specifically volume and/or minimum cross-sectional area (CSA), obtained from computed tomography or magnetic resonance imaging. Additional searches were conducted on the references of included articles and in the NLM catalogue. An assessment of the risk of bias was performed. A total of 1180 studies were retrieved, of which 28 met the eligibility criteria; one was later excluded as it presented a high risk of bias. A meta-analysis was performed. There is moderate evidence to conclude that the upper airway minimum CSA increases significantly (124.13 mm(2)) after maxillomandibular advancement (MMA); the total volume increases significantly after MMA (7416.10mm(3)) and decreases significantly after maxillary advancement+mandibular setback (-1552.90 mm(3)) and isolated mandibular setback (-1894.65 mm(3)).


Direct Oral Anticoagulation Versus Warfarin in Left Ventricular Thrombus: Pooled Analysis of Randomized Controlled Trials.
Patients with impaired left ventricular (LV) function can develop LV thrombus, a potentially life-threatening condition due to risk of stroke and embolization. Conventional treatment with vitamin K antagonists (VKAs; e.g., warfarin) puts patients at risk of bleeding, and the use of direct oral anticoagulants (DOACs) appears promising, although data are scant. We searched the published English language literature for randomized controlled trials (RCTs) comparing DOACs with VKAs in LV thrombus. End points were failure to resolve, thromboembolic events (stroke, embolism), bleeding, or any adverse event (composite of thromboembolism or bleeding), or all-cause death. Data were pooled and analyzed in hierarchical Bayesian models. In three eligible RCTs, 141 patients were studied during an average of 4.6 months (53.8 patient-years; n = 71 assigned to DOAC, n = 70 assigned to VKA). A similar number of patients in each treatment arm demonstrated failure to resolve (DOAC: 14/71 vs. VKA: 15/70) and death events (3/71 vs. 4/70). However, patients on DOACs suffered fewer strokes/thromboembolic events (1/71 vs. 7/70; log odds ratio [OR], -2.02 [95% credible interval (CI95 ), -4.53 to -0.31]) and fewer bleeding events (2/71 vs. 9/70; log OR, -1.62 [CI95 , -3.43 to -0.26]), leading to fewer patients on DOACs with any adverse event versus VKAs (3/71 vs. 16/70; log OR, -1.93 [CI95 , -3.33 to -0.75]). In conclusion, pooled analysis of RCT data favors DOACs over VKAs in patients with LV thrombus in terms of both efficacy and safety.


Long-term outcome by risk factors using conformal high-dose-rate brachytherapy (HDR-BT) boost with or without neoadjuvant androgen suppression for localized prostate cancer.
The aim of this study is to analyze, during the prostate-specific antigen (PSA) era, the long-term outcome of patients treated with conformal high-dose-rate (HDR) brachytherapy boost to the prostate with or without androgen deprivation therapy (ADT) when patients are stratified by risk factors for failure. Between 1986 and 2000, 611 patients were treated for clinically localized prostate cancer in three prospective trials of external beam radiation therapy (EBRT) and dose-escalating HDR brachytherapy (BT) boost. There were 104 patients treated at Seattle, 198 at Kiel University, and 309 at William Beaumont Hospital. Of the 611 patients, 177 received a short course of neoadjuvant/concurrent ADT. The patients were divided into three risk groups. Group I, comprised of 46 patients, had stage < or =T2a, Gleason score (GS) < or = 6, and initial PSA (iPSA) < or = 10 ng/mL. Group II comprised 188 patients with stage > or =T2b, GS > or = 7, and iPSA > or = 10, with any one factor higher. Group III included 359 patients with any two risk factors higher. The American Society for Therapeutic Radiology and Oncology definition for biochemical failure was used. The mean follow-up was 5 years (range, 0.2-15.3). For the 611 patients, the 5-year and 10-year biochemical control (BC) rates were 77% and 73%, disease-free survival (DFS) was 67% and 49%, and cause-specific survival (CSS) was 96% and 92%, respectively. BC at 5 years for Group I patients was 96%, for Group II 88%, and for Group III patients 69%. CSS at 5 years was 100% in Group I, 99% in Group II, and 95% in Group III patients. In univariate and multiple regression analyses for BC, risk group, stage, iPSA, and GS were significant in predicting failure. However, age, follow-up interval, and ADT did not. EBRT with HDR-BT produced excellent long-term outcomes in terms of BC, DFS, and CSS in patients with prostate cancer even for those at highest risk. Conformal HDR-BT is both a precise dose delivery system and an effective treatment for both favorable and unfavorable prostate cancer. The addition of a short course of neoadjuvant/concurrent ADT failed to improve outcome. The results were similar at all three institutions, giving credence to the reproducibility of the brachytherapy treatment.


Validation of a new test for Schistosoma haematobium based on detection of Dra1 DNA fragments in urine: evaluation through latent class analysis.
Diagnosis of urogenital schistosomiasis in chronically infected adults is challenging but important, especially because long term infection of the bladder and urinary tract can have dire consequences. We evaluated three tests for viable infection: detection of parasite specific DNA Dra1 fragments, haematuria and presence of parasite eggs for sensitivity (Se) and specificity (Sp). Over 400 urine specimens collected from adult volunteers in an endemic area in Western Nigeria were assessed for haematuria then filtered in the field, the filter papers dried and later examined for eggs and DNA. The results were stratified according to sex and age and subjected to Latent Class analysis. Presence of Dra1 in males (Se=100%; Sp=100%) exceeded haematuria (Se=87.6%: Sp=34.7%) and detection of eggs (Se=70.1%; Sp=100%). In females presence of Dra1 was Se=100%: Sp=100%, exceeding haematuria (Se=86.7%: Sp=77.0%) and eggs (Se=70.1%; Sp=100%). Dra1 became undetectable 2 weeks after praziquantel treatment. We conclude detection of Dra1 fragment is a definitive test for the presence of Schistosoma haematobium infection.


Preventing worker burnout in child welfare.
Awareness is growing that worker burnout is a serious problem in the child welfare field. This paper examines the multiple causes of the phenomenon, and suggests approaches to prevention.


Nutritional Supplementation With Physical Activity Improves Muscle Composition in Mobility-Limited Older Adults, The VIVE2 Study: A Randomized, Double-Blind, Placebo-Controlled Trial.
Nutritional supplementation and physical activity have been shown to positively influence muscle mass and strength in older adults. The efficacy of long-term nutritional supplementation in combination with physical activity in older adults remains unclear. Mobility-limited (short physical performance battery [SPPB] ≤9) and vitamin D insufficient (serum 25(OH) D 9-24 ng/mL) older adults were recruited for this study. All subjects participated in a physical activity program. Subjects were randomized to consume a daily nutritional supplement (150 kcal, 20 g whey protein, 800 IU vitamin D, 119 mL beverage) or placebo (30 kcal, nonnutritive, 119 mL). In a prespecified secondary analysis, we examined total-body composition (dual energy X-ray absorptiometry), thigh composition (computed tomography), and muscle strength, power, and quality before and after the 6-month intervention. One hundred and forty-nine subjects were randomized into the study [mean (standard deviation, SD) age 78.5 (5.4) years; 46.3% female; mean (SD) short physical performance battery 7.9 (1.2); mean (SD) vitamin D 18.7 (6.4) ng/mL]. After the intervention period both groups demonstrated improvements in muscle strength, body composition, and thigh composition. Nutritional supplementation lead to further losses of intermuscular fat (p = .049) and increased normal muscle density (p = .018). Six months of physical activity resulted in improvements in body composition, subcutaneous fat, intermuscular fat, and strength measures. The addition of nutritional supplementation resulted in further declines in intermuscular fat and improved muscle density compared to placebo. These results suggest nutritional supplementation provides additional benefits to mobility-limited older adults undergoing exercise training. ClinicalTrials.gov Identifier: NCT01542892.


Stereoselectivity of cytochrome P-450 isozymes and epoxide hydrolase in the metabolism of polycyclic aromatic hydrocarbons.
Enantiomeric compositions of epoxides formed in the metabolism of planar benz[a]anthracene (BA), benzo[a]pyrene (BaP), and chrysene (CR), and nonplanar benzo[c]phenanthrene (BcPh), 12-methylbenz[a]anthracene (12-MBA) and 7,12-dimethylbenz[a]anthracene (7,12-DMBA) by liver microsomes from untreated, phenobarbital-treated, and 3-methylcholanthrene-treated rats are determined either by direct chiral stationary phase HPLC analysis or by the enantiomeric compositions of metabolically formed trans-dihydrodiols. Cytochrome P-450 isozymes contained in various liver microsomal preparations have varying degrees of stereoselectivity in catalyzing the epoxidation reactions at various formal double bonds of the polycyclic aromatic hydrocarbons studied. In general, cytochrome P-450c, the major cytochrome P-450 isozyme contained in liver microsomes from 3-methylcholanthrene-treated rats, has the highest degree of stereoselectivity. Regardless of absolute configuration, non-K-region epoxides are converted to trans-dihydrodiols by epoxide hydrolase-catalyzed water attack at the allylic carbon. The S-center of K-region S,R-epoxide enantiomers derived from planar BA, BaP and CR is the major site of epoxide hydrolase-catalyzed water attack. In contrast, the R-center of K-region S,R-epoxide enantiomers derived from nonplanar BcPh, 12-MBA and 7,12-DMBA is the major site of epoxide hydrolase-catalyzed water attack. However, the K-region R,S-epoxide enantiomers of the six polycyclic aromatic hydrocarbons studied are hydrated by microsomal epoxide hydrolase with varying degrees of regioselectivity. Thus the enantiomeric composition of a metabolically formed dihydrodiol is determined by (i) the stereoselective epoxidation at a formal double bond of a parent hydrocarbon by microsomal cytochrome P-450 isozymes and (ii) the enantioselective and regioselective hydration of the metabolically formed epoxide by microsomal epoxide hydrolase.


Distinct guanine nucleotide binding protein alpha-subunit receptor coupling in GH cell lines: effects of bromocriptine and hormones on effector enzyme modulation.
The purpose of the present study was to elucidate how the dopamine agonist bromocriptine affected receptor-effector systems in GH cells by measuring adenylate cyclase (AC) and phospholipase C (PL-C) modulation in cell membrane preparations. To perturb the interaction between the receptor and G-protein, polyclonal antibodies reacting with the predicted C-terminal amino acid sequence of G-protein alpha-subunits were used. The effect of bromocriptine on secretagogue elicited prolactin (PRL) secretion from whole cells was also monitored. Bromocriptine inhibited the basal secretion of PRL in a dose dependent manner, and completely abolished both the thyroliberin (TRH) and the vasoactive intestinal peptide (VIP) stimulated PRL secretion in GH(3) cells. Maximal inhibitory effect on PRL egress elicited by both hormones was obtained at 10-50 microM of bromocriptine. Messenger RNAs for both the short and long form of the D(2) receptor (D(2)R) were demonstrated in all three GH cell lines using the RT-PCR technique, advocating that D(2)Rs are coupled to distinct G-proteins and, thus, probably being responsible for the observed effects of bromocriptine in these cell lines. Basal AC activity, as measured in membrane preparations of GH(3) cells, remained unaffected by bromocriptine treatment (10 microM), while TRH and VIP stimulated AC activities (175% and 350% of control values, respectively) were partially inhibited (by some 50%). This inhibitory effect of bromocriptine was completely and specifically abolished in the presence of an antiserum against G(i2)alpha. Basal PL-C activity was also unaffected by bromocriptine, while TRH stimulated PL-C activity (350% of control value) was inhibited by bromocriptine (10 microM) by approximately 50%. Immunoblocking of G(q/11)alpha, however, reduced the stimulatory effect of TRH on PL-C activation by some 65%, while an antiserum against G(o)alpha partly counteracted the inhibitory effect of bromocriptine (10 microM) on TRH stimulated PL-C activity. Thus, TRH dependent AC stimulation was counteracted by bromocriptine via G(i2). TRH activation of PL-C occurs via G(q/11), while inhibition by bromocriptine appears to involve G(o). These mechanisms probably account for the major part of the actions of bromocriptine, however, other not yet recognised intermediates may be involved.