Assessment of DNA methylation profiling and copy number variation as indications of clonal relationship in ipsilateral and contralateral breast cancers to distinguish recurrent breast cancer from a second primary tumour.
Patients with breast cancer have an increased risk of developing subsequent breast cancers. It is important to distinguish whether these tumours are de novo or recurrences of the primary tumour in order to guide the appropriate therapy. Our aim was to investigate the use of DNA methylation profiling and array comparative genomic hybridization (aCGH) to determine whether the second tumour is clonally related to the first tumour. Methylation-sensitive high-resolution melting was used to screen promoter methylation in a panel of 13 genes reported as methylated in breast cancer (RASSF1A, TWIST1, APC, WIF1, MGMT, MAL, CDH13, RARβ, BRCA1, CDH1, CDKN2A, TP73, and GSTP1) in 29 tumour pairs (16 ipsilateral and 13 contralateral). Using the methylation profile of these genes, we employed a Bayesian and an empirical statistical approach to estimate clonal relationship. Copy number alterations were analysed using aCGH on the same set of tumour pairs. There is a higher probability of the second tumour being recurrent in ipsilateral tumours compared with contralateral tumours (38 % versus 8 %; p <0.05) based on the methylation profile. Using previously reported recurrence rates as Bayesian prior probabilities, we classified 69 % of ipsilateral and 15 % of contralateral tumours as recurrent. The inferred clonal relationship results of the tumour pairs were generally concordant between methylation profiling and aCGH. Our results show that DNA methylation profiling as well as aCGH have potential as diagnostic tools in improving the clinical decisions to differentiate recurrences from a second de novo tumour.


Transmission of wildebeest-associated and sheep-associated malignant catarrhal fever to hamsters, rats and guinea-pigs.
Wildebeest-derived malignant catarrhal fever (WD-MCF) was transmitted to hamsters, rats and guinea-pigs by inoculation of rabbit lymphoid cells infected with alcelaphine herpesvirus-1, strain C-500. Sheep-associated MCF (SA-MCF) was transmitted to hamsters by inoculation of lymphoid cells from rabbits affected with SA-MCF derived from deer. Mice were refractory to both forms of the disease. With both forms of MCF, the incubation period during initial transmission varied from 21 to 90 days and disease was readily passaged in rodents by inoculation of live lymphoid cells. Clinical signs in hamsters most closely resembled those described for naturally occurring MCF. Results given here and in two following papers indicate that rodents are useful models to study the aetiology and pathogenesis of both forms of MCF.


The effects of tamsulosin and alfuzosin on iris morphology: an ultrasound biomicroscopic comparison.
It is well known that Alpha-1 adrenergic receptor antagonists affect the receptors in the prostate and also iris dilator muscle, leading to loss of iris muscle tone. To compare morphological alterations of iris secondary to tamsulosin and alfuzosin use. Patients included in the study were grouped as follows: 16 patients treated with tamsulosin (Group 1), 14 patients treated with alfuzosin (Group 2) and 18 untreated controls (Group 3). All patients underwent ultrasound biomicroscopic and pupillometric examination. Iris thickness was measured at the dilator muscle region (DMR; measured at half of the distance between the scleral spur and the pupillary margin) and sphincter muscle region (SMR; Standardized at 0.75 mm from the pupillary margin). DMR/SMR was also calculated for each patient. Differences among groups were analysed. Main outcome measures were DMR, SMR, DMR/SMR and pupillary diameter. Mean duration of treatments were 2.4 ± 0.96 years (1-4) and 2.3 ± 1.01 years (1-4) in Groups 1 and 2. Pupillary diameters were reduced in Groups 1-2 compared to Group 3 (p < 0.001, p < 0.001). The SMR was similar in Groups 1 and 2 (p: 0.114). These values were not significantly different from that of Group 3 (p: 0.196, p: 0.209). However, thickness in the DMR in Groups 1-2 were significantly lower than that of controls (Group 3) whereas there was no significant difference between Groups 1 and 2 (p: 0.041, p: 0.039 and 0.986, respectively). Mean DMR/SMR ratios were significantly lower in Groups 1-2 than that of Group 3 (p: 0.040 and p: 0.040, respectively). In patients using these medications, the iris seems to be thinner at the dilator muscle region, but preserving the sphincter muscle region.


Two novel HLA-B variants identified in Russian individuals, HLA-B*56:74 and -B*58:124.
HLA-B*56:74 and -B*58:124, two novel alleles detected in Russian individuals by next-generation sequencing.


Acute Kidney Injury and Urinary and Histopathological Disorders in Kidney Transplant Patients with SARS-CoV-2 Infection.
Acute kidney injury (AKI) is a manifestation of SARS-CoV-2 infection. The evidence in kidney transplant (KT) is limited, as there are scarce data about the histologic features in graft biopsies of these patients. A retrospective cohort study of KTs with SARS-CoV-2 infection from August 28, 2020, to April 23, 2021. We collected the incidence of AKI and the presence of urinary and histopathological disorders. Both groups were compared (AKI vs no AKI). Immunohistochemical and reverse transcription-polymerase chain reaction studies were performed on the anatomopathological samples. In our study, 72 KTs had SARS-CoV-2 infection and, among them, 27 patients (35.1%) developed AKI related to increased severity and a worse evolution of the infection, defined by a greater presence of pneumonia (P < .001), hospitalization (P < .001), admission to the intensive care unit (P < .001), the need for ventilation support (P < .001), and continuous renal replacement therapy (P < .001). In the multivariable analysis, pneumonia behaved as an independent predictor for AKI development (P = .046). No differences were observed between proteinuria a month before and after infection (P = .224). In addition, 5 patients showed microhematuria and 2 patients presented transient glycosuria without hyperglycemia. Of the 5 kidney biopsies performed, 1 biopsy (20%) showed positive reverse transcription polymerase chain reaction for SARS-CoV-2. AKI is a frequent and potentially serious complication in KT patients. Occasionally it could be accompanied by abnormalities in the urinary sediment. Of 5 biopsied patients, 1 patient had positive reverse transcription polymerase chain reaction in renal tissue, which suggests the systemic spread of the virus and the tropism for the renal graft.


Risk Factors for Early Cytomegalovirus Reactivation and Impact of Early Cytomegalovirus Reactivation on Clinical Outcomes after T Cell-Replete Haploidentical Transplantation with Post-Transplantation Cyclophosphamide.
Risk factors for cytomegalovirus (CMV) reactivation and the impact of CMV reactivation on patient outcomes have been extensively investigated after matched related or unrelated donor transplantation, but little is known in the setting of haploidentical stem cell transplantation (Haplo-SCT) with post-transplantation cyclophosphamide (PT-Cy), in which recipients are considered more severely immunocompromised. We retrospectively analyzed a cohort of 554 consecutive patients undergoing Haplo-SCT with PT-Cy at 3 different centers. Early CMV reactivation (occurring within the first 120 days post-transplantation) occurred in 242 patients, for an estimated cumulative incidence of 44%. Among those patients, 74 (30%) had recurrent CMV and 20 (8%) had CMV disease. On multivariable analysis, positive recipient CMV serostatus (hazard ratio [HR] >2.5; P < .001), disease histology (lymphoid versus myeloid: HR, 0.66; P = .003) and increasing recipient age (HR, 1.01; P = .015) were independent predictors of CMV reactivation. At a 4-month landmark analysis, CMV reactivation was associated with higher 1-year and 5-year cumulative incidence of nonrelapse mortality (NRM) relative to patients without reactivation: 13% versus 5% and 22% versus 9%, respectively (P < .001). On multivariable analysis, CMV reactivation was an independent negative predictor of NRM (HR, 2.69; P < .001) and was close to statistically significant for overall survival (HR, 1.38; P = .062). Our results suggest that CMV reactivation plays an important role at determining NRM. Because patient CMV serostatus is the main predictor of CMV reactivation, it should be considered when evaluating strategies for preventing CMV reactivation. 2022 American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc.


Azepinoindole derivatives with high affinity for brain dopamine and serotonin receptors.
We synthesized 20 and 21 as conformationally constrained analogues of the dopamine receptor antagonist SKF-83742, as well as analogues 6-9, 16, and 18-22. Although 20 and 21 were inactive, 7, 9, and 19 showed strong binding to D-1, D-2, S-2, and alpha-1 receptors, as well as antipsychotic activity in vivo.


Crop pathogen emergence and evolution in agro-ecological landscapes.
Remnant areas hosting natural vegetation in agricultural landscapes can impact the disease epidemiology and evolutionary dynamics of crop pathogens. However, the potential consequences for crop diseases of the composition, the spatial configuration and the persistence time of the agro-ecological interface - the area where crops and remnant vegetation are in contact - have been poorly studied. Here, we develop a demographic-genetic simulation model to study how the spatial and temporal distribution of remnant wild vegetation patches embedded in an agricultural landscape can drive the emergence of a crop pathogen and its subsequent specialization on the crop host. We found that landscape structures that promoted larger pathogen populations on the wild host facilitated the emergence of a crop pathogen, but such landscape structures also reduced the potential for the pathogen population to adapt to the crop. In addition, the evolutionary trajectory of the pathogen population was determined by interactions between the factors describing the landscape structure and those describing the pathogen life histories. Our study contributes to a better understanding of how the shift of land-use patterns in agricultural landscapes might influence crop diseases to provide predictive tools to evaluate management practices.


Do dorsal head contact forces have the potential to identify impairment during graded craniocervical flexor muscle contractions?
To determine if the force exerted onto the supporting surface by the dorsal head during graded contractions of isometric craniocervical flexion (ICCF) in the supine position has the potential to measure aberrant muscle performance between participant groups with and without painful neck disorders. Cross-sectional, between-participant study of the force exerted by the head on the supporting surface during ICCF muscle contractions. Research laboratory. Thirty-two participants with a history of neck pain and 32 asymptomatic control participants. Not applicable. Dorsal head force (DHF), which is the force (in newtons) exerted by the head on the supporting surface during ICCF muscle contractions in the supine position. ICCF muscle contractions were performed at maximal voluntary contraction (MVC), and sustained at contractions of 50% of MVC and 20% of MVC. Both neck pain and control participants significantly increased their DHF on the supporting surface from resting values during all ICCF muscle contractions (P<.001). No differences in DHF were found between symptomatic and control participants during any of the ICCF efforts. It would appear that DHF exerted on the supporting surface during graded ICCF muscle contractions is similar between neck pain sufferers and control participants, and is therefore limited in its usefulness as a measurement of abnormal performance.


Progression of Pentosan Polysulfate Sodium Maculopathy in a Prospective Cohort.
To describe the progression of pentosan polysulfate sodium (PPS) maculopathy after drug discontinuation qualitatively and quantitatively using multimodal imaging assessmen. Prospective case series. Patients with PPS maculopathy were evaluated after discontinuation of PPS. Near-infrared reflectance (NIR), fundus autofluorescence (FAF), and optical coherence tomography (OCT) were evaluated in all patients at baseline and at the final follow-up visit at least 12 months later. A qualitative and quantitative analysis of the retinal imaging findings was performed. Patterns of disease progression were evaluated. Area of disease involvement on FAF, retinal pigment epithelium (RPE) atrophy on FAF and NIR, and retinal layer thicknesses on OCT were measured at baseline and at the follow-up visit. A total of 26 eyes were included, with a follow-up period ranging from 13 to 30 months. The diseased area measured on FAF showed significant expansion in all eyes from baseline to follow-up despite drug cessation (P = .03) with a median linearized rate of change of 0.42 mm/y. There was significant reduction in the central macular thickness (P = .04), inner nuclear layer thickness (P = .003), outer nuclear layer thickness (P = .02), and subfoveal choroidal thickness (P = .003) at follow-up vs baseline. New areas of RPE atrophy on FAF in the macula developed in 4 eyes while preexisting atrophic lesions increased in size in 5 eyes. Eyes with baseline PPS maculopathy all exhibited remarkable progression with qualitative and quantitative multimodal imaging analysis despite drug discontinuation. Disease progression may be attributed to underlying inner choroidal ischemia or RPE impairment.


A double-labelling study of reticular collaterals to the spinal cord and cerebellum of the North American opossum.
Injections of horseradish peroxidase into either the spinal cord or cerebellum label neurons in the gigantocellular and lateral reticular nuclei of the North American opossum. In order to determine if neurons which project to the spinal cord and cerebellum are intermingled in these two nuclei and if single neurons provide collaterals to both areas, we have employed fluorescent markers in double-labelling experiments. Our results show that reticular neurons innervating either the spinal cord or cerebellum are often close together and that a few provide collaterals to both areas. Neurons providing such collaterals are rare, however, comprising 2% or less of those innervating either target alone.


Outcome from molecular adsorbent recycling system (MARS) liver dialysis following drug-induced liver failure.
Fulminant liver failure from drug ingestion is associated with a high mortality, and the introduction of liver transplantation has improved the mortality significantly if done in a timely fashion. Recently, molecular adsorbent recycling system (MARS) liver dialysis has been introduced as a support for liver failure with varying results. We review our experience with drug-induced liver failure and the impact of MARS liver dialysis on the outcome, in a setting where cadaveric liver transplantation is rarely available. A total of 13 patients were treated, and 40 sessions of MARS liver dialysis were conducted in the intensive care unit. The majority of cases were because of herbal medicine toxicity. Total bilirubin, conjugated bilirubin, and delta bilirubin were significantly reduced, with no change in unconjugated bilirubin. All patients satisfied the criteria for urgent liver transplantation with an average Model End Stage Liver Disease (MELD) score of 35. Only one patient received a liver transplantation from a live donor (right lobe). Overall mortality was 85%. Median time-to-death from the start of MARS was 8 days. MARS liver dialysis in a setting without timely liver transplantation is associated with a poor outcome. It does, however, provide a window of time for consideration of living donors in the setting of limited cadaveric donors.


Medical student education: what it costs and how it is funded.
The cost to Sydney University to educate a medical student in 2010 was obtained by dividing the total teaching costs by the number of students. This showed the annual cost to educate one medical student was $53,093. Sixty-one per cent comprised salaries. Fifty-three per cent is met by Commonwealth funding and the Higher Education Loan Programme. Fees from international medical students contributed a margin of only 2% towards total education costs. The shortfall of 45% is provided from other sources within the university. This cross-subsidisation may not be sustainable.


The effect of antiplatelet drug on coronary endothelial and microvascular function: comparison with ticagrelor and clopidogrel.
Coronary endothelial and microvascular function play important roles in cardiovascular disease. We aimed to evaluate the effect of ticagrelor on coronary artery function and tested the antiplatelet effect of low dose ticagrelor in East-Asian patients. Sixty-one consecutive patients with non-significant coronary disease were included in the study. Initially, patients were randomized in 1:1:1 ratio to receive drugs: ticagrelor 90 mg twice a day (bid; n = 22), ticagrelor 45 mg bid (n = 19) or clopidogrel 75 mg once a day (qd; n = 20) and then divided into two groups (ticagrelor vs clopidogrel) for evaluation of coronary artery function, and three groups for evaluation of antiplatelet function. Endothelial dysfunction was measured by coronary flow reserve (CFR), and changes in the levels of asymmetric dimethylarginine (ADMA), cluster of differentiation (CD) 40 ligand, and P-selectin. Microvascular function was evaluated as index of microvascular resistance (IMR). Platelet reactivity was assessed by VerifyNow P2Y12 assay. The levels of CFR, ADMA, and CD 40 ligand were not different between the two groups. However, P-selectin was lower in the ticagrelor group compared with clopidogrel group. IMR was significantly lower in the ticagrelor group compared with clopidogrel group (median, 15.0 [interquartile range, 12.0 to 21.0] vs. 47.5 [23.0 to 67.5], p = 0.014). There was significant difference in platelet inhibition among the three groups (ticagrelor 90 mg bid vs. ticagrelor 45 mg bid vs. clopidogrel 75 mg qd; 85.57 ± 47.63 vs. 120.33 ± 51.09 vs. 256.42 ± 55.10, p < 0.001). It is hypothesized that ticagrelor might ameliorate the coronary microvascular function. When compared with clopidogrel, low dose ticagrelor exhibited satisfactory antiplatelet effect in the present study.


Angiotensin-converting enzyme: presence of high activity in choroid plexus of mammalian brain.
The activity of angiotensin-converting enzyme in rat choroid plexus was higher than that of any other organ, being 6--7 times higher than that in lung and more than 50 times higher than in any other region of brain. Rabbit choroid plexus also had high activity of enzyme while that of human choroid plexus was relatively low. The enzyme in rat choroid plexus showed similar biochemical properties to that in other tissues; it was inhibited by the nonapeptide SQ 20,881, by (Sar1-Ala8)-angiotensin II and by EDTA, and required chloride ions for activity. As in other tissues, the choroid plexus enzyme was associated with particulate fractions after differential centrifugation. The corpus striatum and substantia nigra had the highest activities in the various brain regions examined.


Plant growth promoting potential and phylogenetic characteristics of a lichenized nitrogen fixing bacterium, Enterobacter cloacae.
Lichens are complex symbiotic association of mycobionts, photobionts, and bacteriobionts, including chemolithotropic bacteria. In the present study, 46 lichenized bacteria were isolated by conventional and enrichment culture methods on nitrogen-free bromothymol blue (NFb) medium. Only 11 of the 46 isolates fixed nitrogen on NFb and had reduced acetylene. All these 11 isolates had also produced siderophore and 10 of them the IAA. Further, ammonia production was recorded from nine of these nitrogen fixers (NF). On molecular characterization, 16 S rRNA sequencing recorded that, nine NF belonged to Proteobacteria, within Gammaproteobacteria, and were closely related to Enterobacter sp. with a maximum similarity to Enterobacter cloacae. Each one of our NF isolates was aligned closely to Enterobacter pulveris strain E443, Cronobacter sakazakii strain PNP8 and Providencia rettgeri strain ALK058. Notably, a few strains we examined found to possess plant growth promoting properties. This is the first report of Enterobacter sp. from lichens which may be inhabit lichen thalli extrinsically or intrinsically.


[The content of various active and inactive genes in chromatin fractions with different accessibilities of DNA to polycyclic aromatic hydrocarbons].
Nuclear chromatin was separated into four fractions according to solubility in low (0.2 mM MgCl2, 10 mM Tris-HCl, pH 7.6) and high (2 M NaCl) ionic strengths after digestion of nuclear DNA with nucleases. In nuclear matrix DNA the ratio of active to inactive genes was always higher than that in the original total DNA, i.e., 25 times greater in rat liver nuclei. In DNA released from the nuclei into a low ionic strength buffer the active to inactive gene ratio was lower than in total DNA (3.7 times as low in case of rat liver nuclei). The amount of carcinogens in matrix DNA exceeded that of DNA soluble in a low ionic strength buffer (3-4 times in case of rat liver nuclei and 16 times in case of hamster embryo cells). The two other fractions occupied an intermediate position between the above said fractions of DNA. The experimental results suggest that the level of carcinogen-induced modifications may be increased in active genes, including transcribed protooncogenes.


A study of the distribution of 7-ethoxycoumarin O-de-ethylase activity in human placental subcellular fractions.
Human placental homogenates from maternal smokers and non-smokers were fractionated using differential centrifugation techniques. Yields of the various subfractions were determined and their homogeneity assessed using electron microscopic procedures. The distribution and response of 7-ethoxycoumarin O-de-ethylase activity towards inhibition by dimethylsulphoxide, alpha-naphthoflavone and 9-hydroxyellipticine inhibitors in the placental subfractions were investigated. The low yield of microsomal protein obtained following differential centrifugation of placental homogenates (2.5 +/- 0.2 mg protein per g placenta) highlights the extremely refractory nature of human placental tissue towards homogenization. Enzymic studies showed that the majority (75 per cent) of the original O-de-ethylase activity in homogenates from smokers and non-smokers was to be found in the crude nuclear fraction. The 7-ethoxycoumarin O-de-ethylase activity present in both homogenate and crude nuclear preparations from a maternal smoker was found to be inhibited by both alpha-naphthoflavone and 9-hydroxyellipticine to a lesser extent than the O-de-ethylase activity which was present in both mitochondrial and microsomal fractions. While this observation suggests the existence of more than one induced O-de-ethylase activity in the human placenta, the possibility that such differences in inhibitory response may be due to other factors (e.g. inhibitor solubility effects) cannot be excluded. Studies using the above inhibitors also confirmed the results of earlier work by demonstrating that the O-de-ethylase activity in placental homogenates and subfractions from non-smokers is qualitatively different from the O-de-ethylase activities induced as a result of maternal smoking.


Development of a pharmacy referral protocol to enhance interprofessional services in a university-affiliated dental medicine clinic.
To discuss the process of developing pharmacy consult services within a university-affiliated dental clinic. Secondary objectives are to describe the frequency of pharmacy consults provided when comparing the initial to the modified protocol and interventions, over time. A university-affiliated student-driven dental medicine clinic in the Midwest. The dental clinic consists of third- and fourth-year dental students who provide dental services and are supervised by dental faculty. Pharmacy consults are prompted due to patient criteria met on the protocol, patient-specific factors, drug information, or other reasons. Pharmacists are present to provide consults directly with dental students or patients. Since October 2013, pharmacy services have been implemented into a dental clinic. A protocol is established to identify dental patients with high risk medications and medical conditions that may interfere with dental treatment. The initial protocol has been modified to include additional high-risk medications and conditions that the pharmacy team was being consulted for outside the protocol criteria. A retrospective chart review was completed to evaluate the frequency of use of the original pharmacy protocol and the interventions provided. After the protocol was modified to include additional high-risk criteria, a second chart review was completed to assess changes in frequency of protocol use and interventions provided. Pharmacists undergo extensive self-learning to understand the dental-related concerns of the high-risk criteria as well as the dental workflow. The original pharmacy protocol accounted for 42.3% (n = 113) of the overall pharmacy consults (n = 267). After protocol modifications, utilization increased to 76.4% (n = 352, total n = 461). Pharmacists are successfully integrated into a dental medicine clinic to provide services to enhance dental patient care. The approximate 30% increase in the use of the pharmacy consult protocol demonstrated that the modified version was more effective.


Characteristics and settling behaviour of particles from blast furnace flue gas washing.
A lot of particles from iron-making are removed with blast furnace off-gas and routed to the gas cleaning system. As water is used for cleaning the gas, the produced wash water contains a large amount of particles such as valuable Fe and C. However, the presence of zinc prevents recycling. In addition, the high amount of calcium results in uncontrolled scaling. Therefore, the properties of the wash water from scrubber and sludge, from the Finnish metal industry (SSAB Raahe), were evaluated in this study. Size fractionation of wash water revealed that Fe, Zn, Al, Mn, V, Cr and Cd appeared mainly in the larger fractions (>1.2 μm) and Na, Mg, Si, Ni, K, Cu and As appeared mainly in the smaller fractions (<1.2 μm) or in dissolved form. Calcium was found both in the larger fractions and dissolved (∼60 mg/L). Most of the particles in wash water were included in the 1.2-10 μm particle size and were settled effectively. However, a clear benefit was observed when using a chemical to enhance particle settling. In comparison to 2.5 h of settling without chemical, the turbidity was further decreased by about 94%, iron 85% and zinc 50%. Coagulation-flocculation experiments indicated that both low and high molecular weight cationic polymers could provide excellent purification results in terms of turbidity. Calcium should be removed by other methods. The particles in sludge were mostly in the 2-4 μm or 10-20 μm fractions. Further sludge settling resulted in high solids removal.


Nitrogen-Walk Approach to Explore Bioisosteric Replacements in a Series of Potent A2B Adenosine Receptor Antagonists.
A systematic exploration of bioisosteric replacements for furan and thiophene cores in a series of potent A2BAR antagonists has been carried out using the nitrogen-walk approach. A collection of 42 novel alkyl 4-substituted-2-methyl-1,4-dihydrobenzo[4,5]imidazo[1,2-a]pyrimidine-3-carboxylates, which contain 18 different pentagonal heterocyclic frameworks at position 4, was synthesized and evaluated. This study enabled the identification of new ligands that combine remarkable affinity (Ki < 30 nM) and exquisite selectivity. The structure-activity relationship (SAR) trends identified were substantiated by a molecular modeling study, based on a receptor-driven docking model and including a systematic free energy perturbation (FEP) study. Preliminary evaluation of the CYP3A4 and CYP2D6 inhibitory activity in optimized ligands evidenced weak and negligible activity, respectively. The stereospecific interaction between hA2BAR and the eutomer of the most attractive novel antagonist (S)-18g (Ki = 3.66 nM) was validated.


Anti-HD IgM as a marker of chronic delta infection.
The value of anti-HD IgM as a marker of chronic delta infection was evaluated by correlating its presence in serum with that of HD-Ag in liver cells and with the degree of inflammatory activity. Thirty-six patients with HBsAg-positive chronic hepatitis and anti-HD at high titers were studied. Overall, the liver cells of 26 patients contained HD-Ag and 27 were positive for IgM anti-HD. The correlation between both markers was excellent: 25 cases were positive for both serum anti-HD IgM and intrahepatic HD-Ag and 8 were negative for both markers. There was only 1 HD-Ag-positive patient, who was anti-HD IgM-negative. Two patients were anti-HD IgM-positive and HD-Ag-negative. Histological damage was more severe in anti-HD IgM-positive cases than in those negative for this marker (Knodell's index 13.5 vs 11.9, P less than 0.01). We conclude that anti-HD IgM is a good marker of chronic active delta infection.


Jejunal perforation complicating dermatomyositis.
Small bowel perforation is rare in dermatomyositis (DM). However, it is associated with high rates of morbidity and mortality. In line with the SCARE criteria, we describe a case of jejunal perforation for a DM patient (Agha et al., 2018 [1]). A 63-year-old woman had been treated for DM with high dose steroid 1 month prior to the onset of severe abdominal pain. Computed tomography (CT) revealed free air in abdominal cavity and fluid around the small bowel. She was diagnosed with small bowel perforation and underwent emergency surgery. Emergent surgery showed perforated jejunum which was resected. Pathologic reports revealed mesenteric small-vessel vasculitis with a perivascular inflammatory cell infiltration. Perforation of the small bowel for DM patients is rare. However, the early diagnosis of bowel perforation is difficult in DM because it can mimic other gastrointestinal manifestations such as ileus, ischemic colitis and peritonitis. To minimize mortality via an early diagnosis and a timely treatment, it is important to examine the patient's clinical history and employ a proper medical imaging modality such as CT even when lab findings are nonspecific and atypical.


Noonan's syndrome associated with cake kidney.
Noonan's syndrome is an established clinical entity. These cases are often associated with correctable cardiac abnormalities, cryptorchism, and abnormal urinary tract. Herein is described the first case of Noonan's syndrome associated with cake kidney. The very high incidence of renal tract abnormalities in association with this syndrome warrants an excretory urogram in these cases for proper evaluation and management.


Endoscopically assisted malarplasty: L-rotation technique.
Endoscopically assisted malarplasty with a greenstick fracture is a method for preserving the body eminence and improving arch protrusion. However, this technique is inadequate when malar body reduction is required. To compensate for these limitations, we developed a new surgical technique using the endoscope to reduce body protrusion. We enrolled 261 patients who visited our clinic for zygoma reduction from January 2017 to December 2018. Their mean age was 31 (range 17-63) years. After a scalp incision, an incomplete osteotomy was created from the most prominent portion of the zygoma body to the zygomaticomaxillary suture line. A complete osteotomy was performed on the arch. These osteotomies resulted in an L-shaped zygoma segment after tapping the bone with a mallet. Of the 261 patients who underwent our L-rotation technique, 242 also received a corticotomy. Of those patients, 15 underwent a different degree of zygoma reduction on both sides. A floating zygomatic segment occurred in four cases, although no further surgery was required. One patient's zygomatic segment dropped on one side, requiring rigid fixation through the intraoral approach. Most patients were satisfied and there were no specific complications. Endoscopically assisted malarplasty using an L-rotation technique enables the protrusion of both the arch and body to be reduced. The zygoma reduction can be modified based on the location of the incomplete osteotomoy line and the number of corticotomies required.


Treating LUTS in Men with Benign Prostatic Obstruction: A Review Article.
Benign prostatic obstruction (BPO) is a prevalent condition that affects men, primarily toward their old age. The condition is often accompanied by lower urinary tract symptoms (LUTS), which can significantly impair a patient's quality of life and lead to other medical complications. Accurate diagnosis of BPO is essential for effective management of complications secondary to BPO, and treatment plans should be tailored patients, and occasionally according to surgeon experience. As such, this literature review aims to analyze the current available data on male LUTS secondary to BPO by providing a comprehensive overview of relevant studies, as well as the surgical and medical management guidelines from the Canadian Urological Association (CUA), American Urological Association (AUA), and European Association of Urology (EAU). By synthesizing the existing literature, this review purports to summarize the current body of knowledge surrounding BPO and male LUTS, and support healthcare providers in making informed decisions about the management of male LUTS secondary to BPO, ultimately improving patient outcomes and quality of life.


Pharmacological approaches to the management of schizophrenia: 10 years on.
To review the contemporary landscape regarding pharmacological treatments for schizophrenia. Selective literature review. Newer antipsychotic agents include aripiprazole, asenapine, paliperidone, sertindole and ziprasidone. Each has some particular benefits and some shortcomings. Overall treatment efficacy (for positive symptoms at least) has not advanced substantially but some newer agents might have a better profile than older typical agents for negative and cognitive symptoms. Metabolic side effects and hyperprolactinaemia remain a problem with some of the newer agents and appropriate monitoring is required. Whilst newer antipsychotics have been welcome additions to our pharmacological armamentarium, mostly in terms of tolerability, we have still not seen a 'quantum leap' agent brought to market. Mechanisms of action apart from post-synaptic dopamine blockade appear worthy of further investigation in this regard.


A neoepitope-based enzyme immunoassay for quantification of C1-inhibitor in complex with C1r and C1s.
Monoclonal antibodies (MoAb) recognizing neoepitopes exposed on activation products of complement proteins but hidden in the native components have been used for quantification of activated complement. A previously produced and characterized mouse MoAb, recognizing a neoepitope on the human plasma protein C1-inhibitor complexed with its substrates, was used to design an enzyme immunoassay for detection of C1-inhibitor complexed with C1r and C1s. These complexes are indicators of early classical complement pathway activation. The standard was serum activated with heat aggregated IgG defined to contain 1000 arbitrary units (AU)/ml. The lower detection limit was approximately 0.05 AU/ml corresponding to 0.005% of fully activated serum. The reliability of the assay, including day-to-day variation, was tested. Intra-assay variation coefficients were 12% for low plasma control and 13% for high plasma control (n = 12 for both). Inter-assay variation coefficients were 12% for low control (n = 6), 19% for high control (n = 6) and 15% for the normal plasma control (n = 9). A 2.5-97.5 percentile reference range (normal blood donors) was 16-33 AU/ml. Two patients with systemic lupus erythematosus had considerably elevated plasma levels of the activation product (56 and 62 AU/ml), and six patients with hereditary angioedema had normal plasma levels despite considerably reduced C1-inhibitor concentration. We conclude that the present method is sensitive and reliable for detection of early classical pathway activation and superior to previously published methods by utilizing neoepitope specificity and non-radiolabelled reagents.


Oral administration of TNP-Lactobacillus conjugates in mice: a model for evaluation of mucosal and systemic immune responses and memory formation elicited by transformed lactobacilli.
Safe live vector systems are being developed for oral delivery of antigens. A transformation system for indigenous Lactobacillus species of the gastrointestinal tract is described. Model systems were set up to evaluate immune responses. Orally administered trinitrophenylized (TNP) Lactobacillus were examined for their ability to induce immunological memory formation via determination of specific antibody titres in serum. We demonstrate a direct correlation between the level of systemic memory formation, as revealed by specific anti-TNP IgG serum antibodies, and the TNP substitution ratio of the Lactobacillus suspension used for oral priming. The specific IgG anti-TNP serum titres were comparable to or even higher than the titres of parental intraperitoneally primed animals. These results demonstrate the feasibility of using orally administered antigen-Lactobacillus as a future approach to vaccination.


Involvement of capsaicin-sensitive nerves in paraquat-induced mortality.
Paraquat (PQ), a broad spectrum herbicide, produces severe lung inflammation and necrosis resulting in pulmonary fibrosis and respiratory failure. Tachykinins are peptides released by sensory C fibers and have the ability of influencing respiratory functions and cellular proliferation. To examine whether the damage caused by PQ involves tachykinins, rats were depleted in their content of tachykinins by systemic treatment with capsaicin prior to PQ exposure. The animal subjected to this treatment showed a 3-fold higher viability compared to those treated with PQ alone (75 vs 27%). Depletion of reduced glutathione (GSH) is associated with oxidative stress produced by reactive oxygen intermediates during PQ metabolism. This is considered to be critical in the pathogenesis of lung damage by PQ. PQ treatment induced a significant depletion of GSH during the first days and a similar effect was also observed in the group of capsaicin-pretreated rats. Four weeks after PQ treatment the levels of GSH were similar to controls in rat pretreated or not with capsaicin plus PQ. This may indicate that the reduced levels of GSH may be associated to the toxicity observed in the acute phase, but not of importance in the final PQ-induced mortality. Neutral endopeptidase (NEP) is an enzyme considered to be critical in controlling the levels of tachykinins. Exposure of crude membrane preparations of rat lung to PQ resulted in a dose-dependent inhibition of NEP activity. Since NEP inactivation may occur in lung following a PQ exposure in vivo, the results indicate that during PQ intoxication a more sustained activity of tachykinins may be present, producing effects such as cell proliferation, fluid extravasation and bronchoconstriction. In conclusion, this finding supports the hypothesis that neuropeptides released from capsaicin-sensitive nerves could be involved in the modulation of PQ-induced lung damage.


Murine chemokine CXCL2/KC is a surrogate marker for angiogenic activity in the inflammatory granulation tissue.
A wide range of compounds inhibit formation of new blood vessels in a variety of models, accompanied by decreases in pro-angiogenic cytokines. The authors sought a surrogate marker for the complex process of neovascularization by correlating inhibition of cytokine production with anti-angiogenic effect. Three anti-angiogenic compounds, clotrimazole (120 mg kg(-1) day(-1)), thalidomide (100 mg kg(-1) day(-1)), and rosiglitazone (10 mg kg(-1) day(-1)), were used to inhibit angiogenesis developing over 9 days, in sponges implanted subcutaneously in Swiss mice. Angiogenesis was assessed by hemoglobin content and by histology. Content of cytokines in implants was measured by specific immunoassays and accumulation of neutrophils or macrophages in implants by measuring myeloperoxidase or N-acetylglucosaminidase activity, respectively. These compounds caused equal inhibition of angiogenesis (about 40%). However, implant levels of vascular endothelial growth factor (VEGF), tumor necrosis factor (TNF-alpha) or the macrophage chemoattractant cytokine, CCL2/MCP-1/JE, and accumulation of macrophages were more variably inhibited. Only the neutrophil chemokine, CXCL2/KC, was inhibited equally by the three compounds, in this model. Anti-angiogenic effect was most clearly and closely correlated with levels of the chemokine KC. Thus, measurement of the chemokine KC might provide an adequate surrogate marker for the functional process of neovascularization in our model.


A fast, aqueous, reversible three-day tissue clearing method for adult and embryonic mouse brain and whole body.
Optical clearing methods serve as powerful tools to study intact organs and neuronal circuits. We developed an aqueous clearing protocol, Fast 3D Clear, that relies on tetrahydrofuran for tissue delipidation and iohexol for clearing, such that tissues can be imaged under immersion oil in light-sheet imaging systems. Fast 3D Clear requires 3 days to achieve high transparency of adult and embryonic mouse tissues while maintaining their anatomical integrity and preserving a vast array of transgenic and viral/dye fluorophores. A unique advantage of Fast 3D Clear is its complete reversibility and thus compatibility with tissue sectioning and immunohistochemistry. Fast 3D Clear can be easily and quickly applied to a wide range of biomedical studies, facilitating the acquisition of high-resolution two- and three-dimensional images.


Biochemical characterization of Nostoc sp. exopolysaccharides and evaluation of potential use in wound healing.
Exopolysaccharides (EPS) produced by cyanobacteria are complex biomolecules of anionic nature with potential biomedical applications. In this study, the EPS produced by the Nostoc sp. strains PCC7936 and PCC7413 were characterized and evaluated as a biomaterial for new wound dressings. The addition of acetate ions to the culture medium slightly stimulated EPS production, achieving 1463.1 ± 16.0 mgL-1 (PCC7413) and 1372.1 ± 29.0 mgL-1 (PCC7936). Both EPS presented nine monosaccharide residues and a MW > 1000 kDa. The acetate addition changed the monosaccharide molar percentages. FTIR and DLS results confirmed the anionic nature and the presence of sulfate groups in both EPS, which are determinant features for biomedical applications. Both EPS at 1%(w/v) formed gels in the presence of 0.4%(w/v) FeCl3. Results obtained for MTT assay and wound healing in vitro scratch assay revealed hydrogels biocompatibility and ability to promote fibroblast migration and proliferation that was greater in PCC7936. The Nostoc EPS hydrogels presented promising properties to be applied in the treatment of skin injuries.


Effects of ultrasonic treatment during acid hydrolysis on the yield, particle size and structure of cellulose nanocrystals.
Ultrasonic treatment is useful for the isolation of cellulose nanocrystals (CNCs). However, the effects of ultrasonic treatment on the structure and properties of CNCs have not been fully understood. We statistically analyzed the variations in yields and dimensions of CNCs prepared using acid hydrolysis, with or without ultrasonic treatment, and illustrated these variations by investigating the CNC structures. The results demonstrated that ultrasonic treatment promoted an increase in yields only for short hydrolysis times, while resulting in CNCs with shorter lengths and narrower dimension distributions. Furthermore, a prolonged acid attack under ultrasonic treatment caused a partial dissociation of cellulose hydrogen bond networks in the CNCs, ultimately resulting in the delamination and disorder of the cellulose crystalline structure, thus leading to a decrease in the width and thickness of the CNCs. This work provides more insights into the mechanisms of ultrasonic treatment on the structure and properties of CNCs.


Weissella confusa N17 Derived from Loach (Misgurnus anguillicaudatus) Exhibits Promising for Further Applications in Loach Aquaculture.
The application of probiotics, in aquaculture, is becoming increasingly widespread and have had positive application effects. However, reports of loach-derived probiotics are quite limited. In this study, two representative strains of lactic acid bacteria with excellent traits, namely, Weissella confusa N17 and Lactobacillus saniviri N19, were screened from the intestine of healthy loaches. W. confusa N17 and L. saniviri N19 could inhibit different common various pathogenic bacteria, especially Aeromonas spp., and were sensitive to the most common antibiotics. The survival rate of the two strains exceeded 50% after 4 h of incubation in 10% loach bile. Moreover, the two strains showed significant tolerance to trypsin. Their autoaggregation capacity and hydrophobicity were greater than 30%. In addition, the aggregation ability of both strains was higher than 30% for both A. veronii TH0426 and A. hydrophila TPS. The two strains had a high biofilm-forming ability and strong adhesion to epithelioma papulosum cyprini (EPC) cells. Scanning electron microscopy results showed that the culture supernatants of the two strains had a significantly destructive effect on A. veronii TH0426 and A. hydrophila TPS. Overall, the traits of W. confusa N17 were better than those of L. saniviri N19. Genome sequencing and analysis demonstrated a lack of virulence factor-related or drug resistance-related genes in genome N17. The diet supplemented with the W. confusa N17 strain significantly improved the resistance of loaches to A. veronii infection, and the protection rate reached 57.1%. Therefore, W. confusa N17 exhibits promising for further applications in loach aquaculture.


A computer program for comparing irregular two-dimensional forms.
A shape comparison of two forms requires a procedure that ensures comparability. A computer program has been written to compute a quantitative measure of shape differences between irregular bounded forms. The curve-fitting procedure, utilizing Fourier's series, is particularly well-suited for two-dimensional closed forms. The program provides several alternate methods to eliminate size difference, ensure a common or neutral center for superimposition and provide a rotational orientation which minimizes shape differences. The program contains numerous user-specified options which control input data transformations, algorithmic processing and output. The algorithmic processing provides: (1) generation of Fourier coefficients based on observed data, (2) two methods of producing Fourier coefficients adjusted for positional orientation, (3) two methods of producing Fourier coefficients adjusted for size standardization, (4) two methods of producing Fourier coefficients adjusted for rotational orientation, (5) a cross-correlation technique which rotationally adjusts two superimposed forms to minimize the shape difference, and (6) various standard statistical measures.


PFAPA syndrome: with regard to a case.
PFAPA syndrome (Periodic Fever, Aphtas, Pharyngitis and cervical Adenopathies) is one of the causes of periodic fever in pediatrics and it is characterised by high fever, pharyngitis, cervical adenitis and aphtous stomatitis. Its etiopathogeny is unknown. The diagnosis is clinical and the findings of laboratory are unspecified. One or two doses (1 mg/kg) of oral prednisone are enough for a fast resolution of the clinic. It is a benign syndrome and no sequels have been noticed after its disappearance, usually in four years from its beginning. We present the case of a 10-year-old patient who has been diagnosed of PFAPA syndrome after 3 years and a half of characteristic clinical bouts, with the fulfilment of diagnostic criteria and after having excluded other entities of similar presentation. Periodic episodes of high fever, pharyngitis and cervical adenitis with a bad response to the conventional treatment should alert us to the PFAPA syndrome. The recognition of this entity will help us to improve the diagnostic and therapeutical focusing, lowering also the anxiety that these cases produce.


Transvenous catheter defibrillation: comparison of a half-sinusoidal and trapezoidal waveform.
This study compares transvenous electrode catheters' defibrillation effectiveness when using the trapezoidal waveform and when using the damped half-sinusoidal waveform, at comparable energy levels. Five greyhounds were used in some 272 fibrillation/defibrillation trials. Neither total energy, peak current, nor peak voltage could be absolutely correlated with percent effectiveness for transvenous defibrillation. However, for a given waveform, an increase in delivered energy was associated with a large and statistically significant increase in percent success.


Feasibility and effectiveness of using coronary stents in the treatment of hepatic artery stenoses after orthotopic liver transplantation: preliminary report.
Our aim is to evaluate the feasibility, efficacy, and patency of using coronary stents for the treatment of hepatic artery stenosis after liver transplantation. Hepatic artery stenosis after liver transplantation can be treated using coronary stents. The low rate of complication, high technical success, and 1-year patency rates are encouraging.


Factors associated with decision-making power on family planning utilization among HIV-positive women attending public health facilities in Eastern Ethiopia.
Family planning for HIV-positive women has numerous advantages. However, the need of family planning utilization is challenged by women's nonautonomous decision-making power. Therefore, this study aimed to examine the level and associated factors of decision-making power to utilize family planning among HIV-positive married women. A facility-based cross-sectional study was conducted from March to June 2020 among 363 HIV-positive married women on ART, using systematic random sampling technique. Logistic regression analysis was used to identify variables that affect women's decision-making power on family planning utilization. Statistical significance was declared at p-value < 0.05 with 95% confidence interval and strength of association was reported by adjusted odds ratio. Overall 55.2% (95% CI: 49.9-60.5) of the women had decision-making power on family planning utilization. Women's having good knowledge (AOR: 2.87, 95% CI: 1.52-5.40), favorable attitude (AOR: 1.96, 95% CI: 1.13-3.38), women's getting family planning counseling in ART clinics (AOR: 2.04, 95% CI: 1.16-3.59), women who get integration service of FP and ART (AOR: 1.83, 95% CI:1.07-3.12) were factors independently associated with women decision-making power on family planning utilization. Decision-making power to utilize family planning among married HIV-positive women was low. Factors like poor knowledge about family planning, dissatisfaction with family planning service, not getting counseling about family planning in ART clinics, and not receiving family planning service in ART clinics were independently associated with women's decision-making power on family planning. Infrastructure linked with the health facility, knowledge, and attitudinal factors should all be combined in future family planning programs.


Factors affecting the scientific research ability and the corresponding countermeasures in clinical postgraduates.
Scientific research ability (SRA) is very important for clinical postgraduates. However, the factors affecting students' SRA are constantly changing with the development of medicine. The aim of this study was to investigate the current situation of SRA in clinical postgraduates and exploring the potential factors and the corresponding countermeasures under the background of new medical science. A total of 133 postgraduates (first- or second-year) were investigated by questionnaire in the Second Affiliated Hospital of Fujian Medical University. All results were analyzed by R software. In terms of the SRA, academic-degree postgraduate students (ADPSs) were significantly better than professional-degree postgraduate students (PDPSs) (P = 0.001), the students with scientific research interest were remarkably better than those without scientific research interest (P = 0.004), the students who mastered statistical analysis methods were more prominent than those who did not (P = 0.007), the students with paper-writing skills were obviously superior to those without it (P = 0.003), and the second-year students were notably better than the first-year students (P = 0.003). Stratified analysis by the above factors except the degree type showed no significant difference in the first-year postgraduates. In the second-year postgraduates, the ADPSs were remarkably superior to the PDPSs (P = 0.002), the students with scientific research interest were obviously better than those without scientific research interest (P = 0.014), the students with more time investment in scientific research were more prominent than those with less time investment in scientific research (P = 0.025), the students with paper-writing skills were notably superior to those without it (P = 0.031), and the students with plotting ability were better than those without it (P = 0.013). The important factors affecting the SRA of clinical postgraduates include the degree type, the grade of student, scientific research interest, time investment in scientific research, statistical analysis methods, paper-writing skills, plotting ability. In short, earlier systematic SRA training contributes to the improvement of SRA in clinical postgraduates, especially in PDPSs.


Prostatic involution after intraprostatic injection of cobra toxin.
We evaluated the comparative effects of intraprostatic injection of cobra cardiotoxin D and botulinum toxin type A on prostate structure in the rat model. A total of 18 Sprague-Dawley® rats weighing 500 to 600 gm received a single 0.1 ml injection of saline (6), botulinum toxin type A (6) or the cardiotoxin D (6) component of cobra (Naja naja atra) toxin in the right and left ventral lobes of the prostate. At 14 days the rats were sacrificed. The prostate glands were harvested, weighed and processed for immunohistochemical and morphological studies. Prostate glands injected with cardiotoxin D showed significantly decreased weight compared to that of prostates injected with botulinum toxin type A and the saline control. Prostatic atrophy in the glandular component with flattening of the epithelial lining was seen histologically in rats that received botulinum toxin and cardiotoxin D. Each group injected with cardiotoxin D and botulinum toxin showed a significant increase in the number of apoptotic cells compared with controls while only the botulinum toxin group showed a significant increase in the number of proliferating cells. Only rats injected with botulinum toxin had body weight loss. Our study shows that intraprostatic injection of cobra cardiotoxin D induces prostatic atrophy and leads to a decrease in prostatic weight greater than that of intraprostatic injection of botulinum toxin type A. No systemic effects, such as decreased body weight, were noted after cardiotoxin D injection. Further studies are warranted but the statistically significant decrease in the number of proliferating cells implies a prolonged effect of cardiotoxin D.


Improving temporal stability of stable cavitation activity of circulating microbubbles using a closed-loop controller based on pulse-length regulation.
Stable cavitation (SC) has shown great potential for novel therapeutic applications. The spatiotemporal distribution of the SC activity of microbubbles circulating in a target region is not only correlated with the uniformity of treatment, but also with some undesirable effects. Therefore, it is important to achieve controllable and desirable SC activity in target regions for improved therapeutic efficiency and biosafety. This study proposes a closed-loop feedback controller based on pulse length (PL) regulation to improve the temporal stability of SC activity. Microbubbles circulating in a physiological flowing phantom were exposed to a 1 MHz focused transducer. The SC signals produced were initially received by another 7.5 MHz plane transducer, followed by high-speed signal acquisition and real-time processing. Based on the real-time-measured SC intensity excited by the current acoustic pulse, the proposed closed-loop feedback controller used three proportional coefficients to regulate the peak negative pressure (PNP) and PL of the next acoustic pulse during the acceleration and stable stages, respectively. The results show that the rise time and the temporal stability of the SC intensity of the microbubbles circulating in these two stages were improved significantly by the optimized proportional coefficients used in the proposed controller. Importantly, when compared with the traditional closed-loop feedback controller based on PNP regulation, the proposed closed-loop feedback controller based on PL regulation reduced the probability of a transition between stable and inertial cavitation, thus avoiding the risk of disadvantageous bioeffects in practical applications. These results demonstrate the effectiveness of the proposed PL-based closed-loop feedback controller and provide a feasible strategy for realization of controllable cavitation activity in applications.


Definitive guiding flange prosthesis: A definitive approach in segmental mandibulectomy defect.
Mandibular discontinuity defects following a segmental mandibulectomy defects present a major challenge to the rehabilitation team. With no immediate intervention to rehabilitate the patient, definitive mandibular guidance prostheses with a metal guiding flange and acrylic teeth on the resected side can be used successfully to stabilize the occlusion and correct the deviation. The present case report describes the prosthodontic rehabilitation of a patient with a segmental mandibulectomy using a mandibular prosthesis with a metal guide flange and a maxillary stabilizing metal framework.


Lipoprotein profiles in human heterozygote carriers of a functional mutation P297S in scavenger receptor class B1.
The scavenger receptor class B type 1 (SR-B1) is an important HDL receptor involved in cholesterol uptake and efflux, but its physiological role in human lipoprotein metabolism is not fully understood. Heterozygous carriers of the SR-B1(P297S) mutation are characterized by increased HDL cholesterol levels, impaired cholesterol efflux from macrophages and attenuated adrenal function. Here, the composition and function of lipoproteins were studied in SR-B1(P297S) heterozygotes.Lipoproteins from six SR-B1(P297S) carriers and six family controls were investigated. HDL and LDL/VLDL were isolated by ultracentrifugation and proteins were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. HDL antioxidant properties, paraoxonase 1 activities, apoA-I methionine oxidations and HDL cholesterol efflux capacity were assessed.Multivariate modeling separated carriers from controls based on lipoprotein composition. Protein analyses showed a significant enrichment of apoE in LDL/VLDL and of apoL-1 in HDL from heterozygotes compared to controls. The relative distribution of plasma apoE was increased in LDL and in lipid-free form. There were no significant differences in paraoxonase 1 activities, HDL antioxidant properties or HDL cholesterol efflux capacity but heterozygotes showed a significant increase of oxidized methionines in apoA-I.The SR-B1(P297S) mutation affects both HDL and LDL/VLDL protein compositions. The increase of apoE in carriers suggests a compensatory mechanism for attenuated SR-B1 mediated cholesterol uptake by HDL. Increased methionine oxidation may affect HDL function by reducing apoA-I binding to its targets. The results illustrate the complexity of lipoprotein metabolism that has to be taken into account in future therapeutic strategies aiming at targeting SR-B1.


Endogenous opiates: through 1978.
The discovery of receptors in the brain for opiates and the structure of the endogenous peptides for these receptors has led to an explosion of interest in this field. The present review is the first of an annual series. It summarizes many of the highlights of research with opiate peptides published with a date of 1978 or earlier.


Wrapping of the omental pedicle flap around esophagogastric anastomosis after esophagectomy for esophageal cancer.
Esophagogastrectomy for esophageal cancer is the standard surgical treatment as a curative measure or for palliation. Esophagogastric anastomotic leakage and stricture are common life-threatening postoperative complications (more so if the leakage occurs in the chest), and the success of the anastomosis created in the reconstruction of the resected esophagus can highly influence morbidity and mortality. A prospective, randomized study was undertaken on 291 patients treated for carcinoma of the esophagus between January 2004 and December 2008. The study excluded 36 patients (12%) who were inoperable. Patients were assigned to 2 treatment groups that consisted of 128 patients in group A and 127 patients in group B according to a restricted, permuted block randomization plan. Patients in group A underwent an esophagogastrectomy with wrapping of the pedicle omental flap around the esophagogastric anastomosis. Group B patients underwent an esophagogastrectomy with only a stapled technique. Of all 255 patients who received an esophagogastric anastomosis, 226 (89%) were discharged from the hospital within 15 days of operation. There was no significant difference between these 2 groups in regard to the incidence of pulmonary complications, abdominal or thoracic infections, and days of hospital stay. Anastomotic leaks occurred in a single patient from group A (1%) and in 7 patients from group B (6%). In group A, 33 patients underwent transhiatal esophagogastrectomy and 95 had thoracic esophagogastrectomy, which resulted in an anastomotic leakage in 1 (3%) and 0 (0%) patients, respectively. In group B, 42 patients had transhiatal esophagogastrectomy and 85 had thoracic esophagogastrectomy, which resulted in anastomotic leakage in 5 (12%) and 2 (2%) patients, respectively. The leakage ratio of group B was significant greater than that of group A (P < .05). Two patients were excluded during the evaluation of the benign stricture due to hospital mortality. Anastomotic strictures were noted in 8 patients from group A (6%) and 20 patients from group B (16%), and the difference in the incidence of anastomotic strictures between these 2 groups was statistically significant (P < .05). Wrapping of the pedicle omental flap around the esophagogastric stapled anastomosis site decreases the incidence of anastomotic leakage and stricture rate after esophagectomy for esophageal cancer, thereby decreasing the morbidity and mortality of the procedure.


High-Throughput Nucleotide Resolution Predictions of Assay Limitations Increase the Reliability and Concordance of Clinical Tests.
The ability of next-generation sequencing (NGS) assays to interrogate thousands of genomic loci has revolutionized genetic testing. However, translation to the clinic is impeded by false-negative results that pose a risk to patients. In response, regulatory bodies are calling for reliability measures to be reported alongside NGS results. Existing methods to estimate reliability do not account for sample- and position-specific variability, which can be significant. Here, we report an approach that computes reliability metrics for every genomic position and sample interrogated by an NGS assay. Our approach predicts the limit of detection (LOD), the lowest reliably detectable variant fraction, by taking technical factors into account. We initially explored how LOD is affected by input material amount, library conversion rate, sequencing coverage, and sequencing error rate. This revealed that LOD depends heavily on genomic context and sample properties. Using these insights, we developed a computational approach to predict LOD on the basis of a biophysical model of the NGS workflow. We focused on targeted assays for cell-free DNA, but, in principle, this approach applies to any NGS assay. We validated our approach by showing that it accurately predicts LOD and distinguishes reliable from unreliable results when screening 580 lung cancer samples for actionable mutations. Compared with a standard variant calling workflow, our approach avoided most false negatives and improved interassay concordance from 94% to 99%. Our approach, which we name LAVA (LOD-aware variant analysis), reports the LOD for every position and sample interrogated by an NGS assay. This enables reliable results to be identified and improves the transparency and safety of genetic tests.


Molecular characterization of HLA class II binding to the LAG-3 T cell co-inhibitory receptor.
Immune checkpoint inhibitors (antibodies that block the T cell co-inhibitory receptors PD-1/PD-L1 or CTLA-4) have revolutionized the treatment of some forms of cancer. Importantly, combination approaches using drugs that target both pathways have been shown to boost the efficacy of such treatments. Subsequently, several other T cell inhibitory receptors have been identified for the development of novel immune checkpoint inhibitors. Included in this list is the co-inhibitory receptor lymphocyte activation gene-3 (LAG-3), which is upregulated on T cells extracted from tumor sites that have suppressive or exhausted phenotypes. However, the molecular rules that govern the function of LAG-3 are still not understood. Using surface plasmon resonance combined with a novel bead-based assay (AlphaScreenTM ), we demonstrate that LAG-3 can directly and specifically interact with intact human leukocyte antigen class II (HLA-II) heterodimers. Unlike the homologue CD4, which has an immeasurably weak affinity using these biophysical approaches, LAG-3 binds with low micromolar affinity. We further validated the interaction at the cell surface by staining LAG-3+ cells with pHLA-II-multimers. These data provide new insights into the mechanism by which LAG-3 initiates T cell inhibition.


Air Bubbles Activate Complement and Trigger Hemostasis and C3-Dependent Cytokine Release Ex Vivo in Human Whole Blood.
Venous air embolism, which may complicate medical and surgical procedures, activates complement and triggers thromboinflammation. In lepirudin-anticoagulated human whole blood, we examined the effect of air bubbles on complement and its role in thromboinflammation. Whole blood from 16 donors was incubated with air bubbles without or with inhibitors of C3, C5, C5aR1, or CD14. Complement activation, hemostasis, and cytokine release were measured using ELISA and quantitative PCR. Compared with no air, incubating blood with air bubbles increased, on average, C3a 6.5-fold, C3bc 6-fold, C3bBbP 3.7-fold, C5a 4.6-fold, terminal complement complex sC5b9 3.6-fold, prothrombin fragments 1+2 (PTF1+2) 25-fold, tissue factor mRNA (TF-mRNA) 26-fold, microparticle tissue factor 6.1-fold, β-thromboglobulin 26-fold (all p < 0.05), and 25 cytokines 11-fold (range, 1.5-78-fold; all p < 0.0001). C3 inhibition attenuated complement and reduced PTF1+2 2-fold, TF-mRNA 5.4-fold, microparticle tissue factor 2-fold, and the 25 cytokines 2.7-fold (range, 1.4-4.9-fold; all p < 0.05). C5 inhibition reduced PTF1+2 2-fold and TF-mRNA 12-fold (all p < 0.05). C5 or CD14 inhibition alone reduced three cytokines, including IL-1β (p = 0.02 and p = 0.03). Combined C3 and CD14 inhibition reduced all cytokines 3.9-fold (range, 1.3-9.5-fold; p < 0.003) and was most pronounced for IL-1β (3.2- versus 6.4-fold), IL-6 (2.5- versus 9.3-fold), IL-8 (4.9- versus 8.6-fold), and IFN-γ (5- versus 9.5-fold). Antifoam activated complement and was avoided. PTF1+2 was generated in whole blood but not in plasma. In summary, air bubbles activated complement and triggered a C3-driven thromboinflammation. C3 inhibition reduced all mediators, whereas C5 inhibition reduced only TF-mRNA. Combined C5 and CD14 inhibition reduced IL-1β release. These data have implications for future mechanistic studies and possible pharmacological interventions in patients with air embolism.


The effects of N-acetylcysteine on cisplatin-induced cardiotoxicity on isolated rat hearts after short-term global ischemia.
The aim of this study was to estimate the protective effect of N-acetyl-l-cysteine (NAC) against cisplatin-induced cardiotoxicity under conditions of ischemic-reperfusion injury. Wistar albino rats were randomly divided into three groups (n = 8): control, cisplatin (5 mg/kg/w, i.p., 5 weeks) and cisplatin + NAC group (cisplatin - 5 mg/kg/w, i.p. and NAC - 500 mg/kg/w, i.p., 5 weeks). Isolated hearts were perfused according to the modified Langendorff technique at constant pressure (70 cmH2O). Following cardiodynamic parameters were measured: maximum rate of left ventricular pressure development, minimum rate of left ventricular pressure development, left ventricular systolic pressure (SLVP), left ventricular diastolic pressure and heart rate. The ischemic vasodilation episodes were induced by the complete interruption of coronary inflow for 30, 60 and 120 s. The samples of the coronary venous effluent (CVE) were continuously collected during the reperfusion period for determination of coronary flow (CF) rate and oxidative stress markers (H2O2, O2-, NO2- and thiobarbituric acid reactive substances - TBARS). Cisplatin reduced CF, heart rate and overflow (total, maximal and duration of overflow) during reperfusion, and increased SLVP (under basal conditions and after global ischemias). Cisplatin increased levels of H2O2 (under basal conditions), O2- and TBARS (under basal conditions and after ischemia), but decreased NO2- levels (during reperfusion) in CVE, and decreased superoxide dismutase and reduced glutathione in serum. NAC attenuated cisplatin-induced changes of cardiodynamic parameters (except CF under basal conditions) and oxidative stress parameters. Those results suggest that NAC, by decreasing oxidative stress, may be useful in cardioprotection during cisplatin therapy.


The Bridge Building Model: connecting evidence-based practice, evidence-based research, public involvement and needs led research.
This paper describes a model developed by an interdisciplinary team of research and public engagement specialists, with backgrounds in health and social care research, higher education, evidence-based practice, leadership, commissioning research and public involvement and engagement. The model we propose combines evidence-based practice, evidence-based research, public involvement and needs led research. Our aim is to capitalise on the joining of the rationale and methods for these approaches, which have all been highlighted as important, but for which there has been a lack of models for integration. Our ambition is to argue for and show an effective and evidence-based way of working that bridges health and social care needs identification, evidence-based practice and research.


Methylxanthine activation of noradrenergic unit activity and reversal by clonidine.
Methylxanthines increase central noradrenergic turnover by an unknown mechanism. Isobutyl methylxanthine (IBMX, 0.5-16 mg/kg i.v.) was found in this study to increase firing rats of noradrenergic neurons in the locus coeruleus, supporting increased impulse flow as one possible mechanism for increased noradrenergic turnover. Naloxone (1 mg/kg i.p.) pretreatment potentiated the effects of IBMX, while clonidine (0.01 mg/kg i.v.) reversed increases in activity. Some of the behavioral effects of methylxanthines are consistent with increased noradrenergic activity.


Vasoactive intestinal peptide: a secretin-like partial agonist for pancreatic secretion in man.
The responses of pancreatic volume flow and bicarbonate output to intravenous vasoactive intestinal peptide (VIP, 0.8 to 3.2 microgram per kg per hr) and synthetic secretin (32.2 to 129 ng per kg per hr) were compared intraindividually in 5 healthy volunteers. Pure pancreatic juice was obtained by endoscopic cannulation of the main pancreatic duct. The mean +/- SEM observed maximal response of secretin-stimulated juice flow was 248 +/- 7 microliter per kg per 5 min, whereas the observed maximal response for VIP-evoked juice flow was 48 +/- 6 microliter per kg per 5 min. The observed maximal secretin-induced bicarbonate output was 30 +/- 2 muEq per kg per 5 min, and the maximal VIP-related response was 4.3 +/- 0.9 muEq per kg per 5 min. In addition to low efficacy, high dose requirements, and side effects (significant rise in pulse rate and cutaneous flushing at 3.2 micrograms per kg per hr) argue against a major physiological role of VIP as a hormonal stimulant of human pancreatic bicarbonate secretion.


Effect of Diets with Different Lipid's Sources On Serum and Brain Fatty Acids Profile: Experimental Model.
The importance of diet in health is widely accepted and recognized. Diet lipid profile is important to prevent chronic diseases and improve the quality of an individual's life. The objective of this report is to analyze the effect of different sources of dietary lipids with standard and high concentration on growing rats. Experimental diets contained 15 or 42% kcal of fat, provided by butter (B), olive oil (O), high oleic sunflower oil (HO), and sunflower oil (S). Control diet (C) was normocaloric with 15% kcal of fat provided by soy oil. All diets were complete in the other nutrients according to AIN 1993 and were administered for 40 days. Daily intake was similar in all groups. The administration of these diets provoked changes in serum fatty acid profile in response to the different sources of dietary lipids used; no changes were observed in the brain´s fatty acids. These results would suggest that the organism tries first to supply the brain´s fatty acid needs at the expense of its modification in serum.


Necroptosis in acute kidney injury: a shedding light.
Acute kidney injury (AKI) is a common and severe clinical condition with a heavy healthy burden around the world. In spite of supportive therapies, the mortality associated with AKI remains high. Our limited understanding of the complex cell death mechanism in the process of AKI impedes the development of desirable therapeutics. Necroptosis is a recently identified novel form of cell death contributing to numerable diseases and tissue damages. Increasing evidence has suggested that necroptosis has an important role in the pathogenesis of various types of AKI. Therefore, we present here the signaling pathways and main regulators of necroptosis that are potential candidate for therapeutic strategies. Moreover, we emphasize on the potential role and corresponding mechanisms of necroptosis in AKI based on recent advances, and also discuss the possible therapeutic regimens based on manipulating necroptosis. Taken together, the progress in this field sheds new light into the prevention and management of AKI in clinical practice.


Study of the release mechanism of diltiazem hydrochloride from matrices based on chitosan-alginate and chitosan-carrageenan mixtures.
The aim of this work was to establish the diltiazem hydrochloride release mechanism from the chitosan-alginate matrix tablet (MCB/AS) and chitosan-carrageenan matrix tablet (MCS/CSI). The weight loss for MCS/CSI is mainly due to the weight loss of the matrix while for MCB/AS it is mainly due to the diltiazem hydrochloride released from the tablet. Using the Peppa's model the release order for MCS/CSI was n = 1.07 +/- 0.13 and for MCB/AS was n = 0.76 +/- 0.02. Thus, MCS/CSI has a transport mechanism, and for MCB/AS the drug release mechanism is a combined process of diffusion and relaxation. MCB/AS has an elastic modulus (G' = 10(5) Pa) one order of magnitude higher than MCS/CSI (G' = 10(4) Pa). MCB/AS is able to uptake solvent without disrupting the microstructure due to its high elastic modulus. Instead MCS/CSI showed a quick erosion process, which conducted to the tablet disintegration due to a fast solvent uptake process.


Intraocular pressure fluctuation during microincision vitrectomy with constellation vision system.
To investigate intraocular pressure (IOP) fluctuation during various vitrectomy maneuvers using the vitrectomy system (Alcon Constellation Vision System). An experimental study as laboratory investigation. In porcine eyes, 23- and 25-gauge vitrectomy was performed, and IOP fluctuations were evaluated in vitreous cutting mode, in aspiration mode, and during scleral compression. The measurements were performed with the IOP control setting turned on or off. Using the 23-gauge system with the IOP control setting turned on, IOP decreased from 30 to 23.7 mm Hg after starting vitreous cutting, and then returned to 30 mm Hg in 2.6 seconds. When the IOP control setting was turned off, IOP decreased to 19.1 mm Hg in 0.9 seconds, and remained at that pressure. Under aspiration at 650 mm Hg without cutting, IOP showed a sharp depression from 30 to 12.2 mm Hg, and then returned to 30.6 mm Hg in 2.6 seconds with the IOP control setting turned on. When the IOP control setting was turned off, IOP decreased to 2.2 mm Hg in 9.7 seconds, and did not recover. When the sclera was compressed without aspiration, IOP rapidly increased to 70-100 mm Hg, and then slowly decreased to 30 mm Hg in 3.5-4.0 seconds, with or without the IOP control system. Similar data were obtained with 25-gauge vitrectomy. The IOP control system can attenuate IOP fluctuations during vitrectomy maneuvers. There was no significant difference in IOP fluctuations between 23- and 25-gauge systems.


Implementation and Performance of DFT-D with Respect to Basis Set and Functional for Study of Dispersion Interactions in Nanoscale Aromatic Hydrocarbons.
The implementation, optimization, and performance of various DFT-D schemes have been tested on models for polar-π interactions between arenes spaced at van der Waals distances and on a series of functionalized corannulene derivatives and complexes. For DFT-D schemes involving a semiempirical correction, optimized parameters are proposed for several basis sets. Performance of the different DFT-D strategies is compared, where functionals include some of the most recently proposed, B97D, B2PLYP, BMK, and M06-2X functionals, together with several other well-known functionals. Semiempircally corrected dispersion functionals hold some promise as useful and affordable methods for studies involving large polynuclear aromatic molecules and molecules on metal surfaces.


Study of volatile compounds in Greek pistachio (Pistacia vera L. 'Aegina' cultivar) oils using Soxhlet and ultrasound assisted extraction.
Headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (HS-SPME/GC-MS) represents the most used solvent-free methodology for the characterization of the complex and heterogeneous mix of volatile compounds. The present study investigates the differences in volatile profile of pistachio oils 'Aegina' cultivar extracted with two different techniques, ultrasound assisted extraction (UAE) and Soxhlet. Differences were observed both in the pistachio oil yield and the composition of the volatile compounds among these two groups of samples, which were significantly influenced due to the different thermal conditions. In terms of pistachio oil yield, the Soxhlet extraction technique was proven more efficient (52.5-68.2% w/w) than the UAE type (28.2-42.6% w/w). A total of 34 and 30 volatile compounds were identified for UAE and Soxhlet, respectively. The main ones associated with UAE were α-pinene, octane and decane, while the volatiles formed as a consequence of Soxhlet extraction were decane, nonanal and (E)-2-decenal. Terpenes' concentrations were found decreased in Soxhlet's samples, but hydrocarbons and aldehydes were significantly increased in these samples. Numerous studies concluded in common results. However, this article is the first to explore the influence of different extraction types on the volatile profile of the unique flavour and odor pistachio oil 'Aegina' cultivar.


Demonstrating equivalence and non-inferiority of medical education concepts.
In medical education, there are often several didactic methods for teaching and learning a specific medical skill. For educators, there are often pragmatic reasons to decide for one or another of them, such as costs, infrastructural requirements, time expenditure or qualification of the teacher. However, a central aspect to consider is the learning outcome: Does a new method achieve a similar learning success as an established standard method? To answer this question, we need an appropriate method to assess comparability of learning outcomes. In this paper, we present two essential statistical concepts that can address the issue of comparability of learning outcomes: Equivalence and non-inferiority testing. We explain the ideas behind these concepts and illustrate them with an example data set. To clarify several concepts, we use theoretical examples from one selected field: the teaching and assessment of Basic Life Support (BLS). Equivalence and non-inferiority tests can be powerful tools for comparing teaching and assessment methods. However, their correct application requires adequate knowledge about their strengths, pitfalls and application fields. The aim of this paper is to deliver this knowledge and to provide clinician researchers with a practical guidance to a successful application of these methods.


Prosthetic mitral valve mucormycosis caused by Mucor species.
We report, to the best of our knowledge the first case of prosthetic mitral valve mucormycosis caused by Mucor species in an 18-year-old female after mitral valve replacement. She was successfully treated with surgery and antifungal treatment. Mucormycosis was documented by direct demonstration, isolation and histopathological findings of the fungus from embolus and prosthetic mitral valve thrombus.


SPDI: data model for variants and applications at NCBI.
Normalizing sequence variants on a reference, projecting them across congruent sequences and aggregating their diverse representations are critical to the elucidation of the genetic basis of disease and biological function. Inconsistent representation of variants among variant callers, local databases and tools result in discrepancies that complicate analysis. NCBI's genetic variation resources, dbSNP and ClinVar, require a robust, scalable set of principles to manage asserted sequence variants. The SPDI data model defines variants as a sequence of four attributes: sequence, position, deletion and insertion, and can be applied to nucleotide and protein variants. NCBI web services convert representations among HGVS, VCF and SPDI and provide two functions to aggregate variants. One, based on the NCBI Variant Overprecision Correction Algorithm, returns a unique, normalized representation termed the 'Contextual Allele'. The SPDI data model, with its four operations, defines exactly the reference subsequence affected by the variant, even in repeat regions, such as homopolymer and other sequence repeats. The second function projects variants across congruent sequences and depends on an alignment dataset of non-assembly NCBI RefSeq sequences (prefixed NM, NR and NG), as well as inter- and intra-assembly-associated genomic sequences (NCs, NTs and NWs), supporting robust projection of variants across congruent sequences and assembly versions. The variant is projected to all congruent Contextual Alleles. One of these Contextual Alleles, typically the allele based on the latest assembly version, represents the entire set, is designated the unique 'Canonical Allele' and is used directly to aggregate variants across congruent sequences. The SPDI services are available for open access at: https://api.ncbi.nlm.nih.gov/variation/v0. Supplementary data are available at Bioinformatics online.


Efficiency of ozonation and O3/H2O2 as enhanced wastewater treatment processes for micropollutant abatement and disinfection with minimized byproduct formation.
Ozonation, a viable option for improving wastewater effluent quality, requires process optimization to ensure the organic micropollutants (OMPs) elimination and disinfection under minimized byproduct formation. This study assessed and compared the efficiencies of ozonation (O3) and ozone with hydrogen peroxide (O3/H2O2) for 70 OMPs elimination, inactivation of three bacteria and three viruses, and formation of bromate and biodegradable organics during the bench-scale O3 and O3/H2O2 treatment of municipal wastewater effluent. 39 OMPs were fully eliminated, and 22 OMPs were considerably eliminated (54 ± 14%) at an ozone dosage of 0.5 gO3/gDOC for their high reactivity to ozone or •OH. The chemical kinetics approach accurately predicted the OMP elimination levels based on the rate constants and exposures of ozone and •OH, where the quantum chemical calculation and group contribution method successfully predicted the ozone and •OH rate constants, respectively. Microbial inactivation levels increased with increasing ozone dosage up to ∼3.1 (bacteria) and ∼2.6 (virus) log10 reductions at 0.7 gO3/gDOC. O3/H2O2 minimized bromate formation but significantly decreased bacteria/virus inactivation, whereas its impact on OMP elimination was insignificant. Ozonation produced biodegradable organics that were removed by a post-biodegradation treatment, achieving up to 24% DOM mineralization. These results can be useful for optimizing O3 and O3/H2O2 processes for enhanced wastewater treatment.


The effects of encapsulation on damage to molecules by electron radiation.
Encapsulation of materials imaged by high resolution transmission electron microscopy presents a promising route to the reduction of sample degradation, both independently and in combination with other traditional solutions to controlling radiation damage. In bulk crystals, the main effect of encapsulation (or coating) is the elimination of diffusion routes of beam-induced radical species, enhancing recombination rates and acting to limit overall damage. Moving from bulk to low dimensional materials has significant effects on the nature of damage under the electron beam. We consider the major changes in mechanisms of damage of low dimensional materials by separating the effects of dimensional reduction from the effects of encapsulation. An effect of confinement is discussed using a model example of coronene molecules encapsulated inside single walled carbon nanotubes as determined from molecular dynamics simulations calculating the threshold energy required for hydrogen atom dissociation. The same model system is used to estimate the rate at which the nanotube can dissipate excess thermal energy above room temperature by acting as a thermal sink.


Financial stress experienced by informal carers of adults with a chronic disease: Results from an Australian population-based cross-sectional survey.
To identify caregiving characteristics that are associated with financial stress in Australian carers of people with a chronic disease. Data were collected via the South Australian Health Omnibus, an annual population-based, cross-sectional survey. Individuals who provided care to someone with prevalent chronic conditions were asked about financial stress and caregiving characteristics. Of 32.4% (988/3047) who were carers, 13.4% (132/988) experienced financial stress. Adjusting for age and household income, providing more than 20 hours of care per week (AOR = 2.39, 95% CI = 1.48-3.86), transport assistance (AOR = 1.89, 95% CI = 1.15-3.09) and assistance with household tasks (AOR = 1.92, 95% CI = 1.14-3.26) and caring for a person with a mental illness (AOR = 2.01, 95% CI = 1.24-3.28) were associated with a significant increase in odds of experiencing financial stress. Caring for a person with cancer (AOR = 0.49, 95% CI = 0.30-0.81) or dementia (AOR = 0.40, 95% CI = 0.21-0.76) was associated with decrease in odds. Financial stress was reported by more than 13% of carers, and factors other than household income were implicated.


The cDNA-deduced primary structure of human sex hormone-binding globulin and location of its steroid-binding domain.
We have sequenced a cDNA for sex hormone-binding globulin (SHBG) isolated from a phage lambda gt11 human liver cDNA library. The library was screened with a radiolabeled rat androgen-binding protein (ABP) cDNA, and the abundance of SHBG cDNAs was 1 in 750,000 plaques examined. The largest human SHBG cDNA (1194 base-pairs) contained a reading frame for 381 amino acids. This comprised 8 amino acids of a signal peptide followed by 373 residues starting with the known NH2-terminal sequence of human SHBG, and ending with a termination codon. The predicted polypeptide Mr of SHBG is 40,509, and sites of attachment of one O-linked (residue 7) and two N-linked oligosaccharide (residues 351 and 367) chains were identified. Purified SHBG was photoaffinity-labeled with delta 6-[3H]testosterone and cleaved with trypsin. The labeled tryptic fragment was isolated by reverse-phase HPLC, and its NH2-terminal sequence was determined. The results suggest that a portion of the steroid-binding domain of SHBG is located between residue 296 and the 35 predominantly hydrophilic residues at the C-terminus of the protein.


Doping control analysis in human urine by liquid chromatography-electrospray ionization ion trap mass spectrometry for the Olympic Games Athens 2004: determination of corticosteroids and quantification of ephedrines, salbutamol and morphine.
A new liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) (n) ion trap method for the determination of corticosteroids in urine has been developed and validated. Some anabolic agents, such as epitrenbolone, trenbolone, 2-hydroxymethylformebolone, tetrahydrogestrinone, gestrinone and formoterol were included in the LC-ESI-MS method. Matrix interference, specificity, identification capability, carry over and robustness were estimated as validation parameters. Recoveries ranged from 74 to 113% at the minimum required performance limit (MRPL), which is 30 ng mL(-1) for corticosteroids and 10 ng mL(-1) for anabolic agents. Methods for the confirmation and quantification of norpseudoephedrine, ephedrine, methylephedrine, salbutamol, morphine and morphine glucuronide were also developed and validated and in order to minimize analysis time, direct urine injection was used. These methods proved to be specific, accurate and precise across a calibration range for each substance since matrix interference, specificity, carry over, within and between run precision, limit of detection, limit of quantification, intermediate precision and uncertainty were estimated.


CaMKII activates ASK1 to induce apoptosis of spinal astrocytes under oxygen-glucose deprivation.
Calcium/calmodulin-dependent protein kinase II (CaMKII) is a ubiquitous, structurally complex multifunctional protein serine/threonine kinase that plays an important role in cell apoptosis via linking the ER stress and mitochondrial apoptosis pathways. Recently, CaMKII has been correlated with apoptosis signal-regulating kinase 1 (ASK1) activity and the ASK1-dependent apoptosis pathway through the direct phosphorylation of Thr845 of ASK1. The specific role of CaMKII in hypoxia-reoxygenation (H/R)-induced spinal astrocyte apoptosis, however, remains unclear. In this study, we investigated the effects of CaMKIIγ (an isoform of CaMKII) on spinal astrocyte apoptosis using an in vitro oxygen-glucose deprivation (OGD/R) model which mimics hypoxic/ischemic conditions in vivo. OGD/R increased cell death and the activation of CaMKII. Deletion of CaMKIIγ results in the reduced activation of CaMKII and apoptosis in astrocytes under OGD/R conditions. Notably, the deletion of CaMKIIγ induced ASK1 phosphorylation at Thr845 in astrocytes. The activation of JNK and p38 and the downstream effect of ASK1 were also reduced. These data suggest that CaMKIIγ is required for the CaMKII-dependent regulation of ASK1, affecting the apoptosis of a biologically important cell type under spinal cord injury.


"It is really so exhausting": Exploring intensive care nurses' perceptions of 24-hour long shifts.
To explore the perceptions and experiences of Cambodian ICU registered nurses regarding their working 24-hr shifts. In Europe and the USA, nurses are moving to a 12-hr shift, and numerous studies have revealed the positive and negative effects of these. However, lesser known is the impact of 24-hr nursing shifts on care quality, and health and safety. In Cambodia, 100% of nurses are rostered for these in their shift patterns, but until this study no research had been conducted on such shifts. A qualitative descriptive study. Three focus group discussions were conducted with 30 registered nurses in July 2019, ten in each group, from three intensive care units of three hospitals in Cambodia. Data saturation was obtained. Data were analysed using content analysis, and the COREQ was applied for reporting this study. The ICU nurses' perspectives revealed significant and unacceptable effects of working shifts of ~25-hr length, taking into account staff handover. Two major themes arose: It is so exhausting and Compromised hospital care. Alarmingly, participants worked on average 72 hr per week, were exhausted, and nursed between 6 and 10 critically ill patients per shift. To our knowledge this is the first study on nurses working 24-hr shifts, revealing unacceptable, high risks for the health and safety of nurses and patients, with nursing activities left undone, and a lack of quality care. Improving nurse and patient health and safety, and quality of care requires hospital leaders to work with government and nursing organisations to develop better shift strategies. Resources need to be provided so that: nurses can work a maximum 12-hr shifts; the ratio of nurses to patients is improved; and nurses can have decent break times. This has major implications, for not only practice, but also management, administration, budgets and education.


OX40L-expressing recombinant modified vaccinia virus Ankara induces potent antitumor immunity via reprogramming Tregs.
Effective depletion of immune suppressive regulatory T cells (Tregs) in the tumor microenvironment without triggering systemic autoimmunity is an important strategy for cancer immunotherapy. Modified vaccinia virus Ankara (MVA) is a highly attenuated, non-replicative vaccinia virus with a long history of human use. Here, we report rational engineering of an immune-activating recombinant MVA (rMVA, MVA∆E5R-Flt3L-OX40L) with deletion of the vaccinia E5R gene (encoding an inhibitor of the DNA sensor cyclic GMP-AMP synthase, cGAS) and expression of two membrane-anchored transgenes, Flt3L and OX40L. Intratumoral (IT) delivery of rMVA (MVA∆E5R-Flt3L-OX40L) generates potent antitumor immunity, dependent on CD8+ T cells, the cGAS/STING-mediated cytosolic DNA-sensing pathway, and type I IFN signaling. Remarkably, IT rMVA (MVA∆E5R-Flt3L-OX40L) depletes OX40hi regulatory T cells via OX40L/OX40 interaction and IFNAR signaling. Single-cell RNA-seq analyses of tumors treated with rMVA showed the depletion of OX40hiCCR8hi Tregs and expansion of IFN-responsive Tregs. Taken together, our study provides a proof-of-concept for depleting and reprogramming intratumoral Tregs via an immune-activating rMVA.


Concussions in NCAA Varsity Football Athletes: A Qualitative Investigation of Player Perception and Return to Sport.
The objective of this study was to understand the experiences and perspectives of varsity football athletes on return to play following a concussion injury. Two experienced interviewers conducted qualitative, semistructured interviews of college-level football players who had sustained at least one concussion during their varsity careers. Twenty varsity football players who averaged 2.3 concussions each (range, 1-5) were interviewed regarding peer pressure, the culture of football, and player awareness as factors affecting return to sport following a concussion. Less common secondary factors included risk management, severity and timing of the injury, and team support. Psychological stressors, the culture of football, and increased awareness were the most influential factors affecting collegiate football players' deciding to return to sport following a concussion. Level III.


Virus-specific memory T cells are Pgp-1+ and can be selectively activated with phorbol ester and calcium ionophore.
Memory lymphocytic choriomeningitis virus (LCMV)-immune cytotoxic T-lymphocyte precursors (CTLp) can be stimulated to proliferate and to mediate specific cytotoxic activity following incubation with phorbol myristate acetate (PMA), calcium ionophore (CaI), and interleukin 2 (IL-2). This protocol can be used to selectively induced virus-specific CTL activity under both bulk culture and limiting dilution conditions, in the absence of added antigen. There is no concurrent stimulation of alloreactive CTLp. Proliferation of the effector Lyt-2+ population in medium containing PMA and CaI requires L3T4+ cells, which can be replaced by adding IL-2, and the development of cytotoxicity is totally IL-2 dependent. The LCMV-specific memory T cells are also characterized by the expression of the Pgp-1 (Ly24) glycoprotein. The availability of this marker, together with the capacity to selectively stimulate primed CTLp in the absence of antigen, should greatly facilitate the analysis of T-cell memory in virus infections.


Relationship between the superprism effect in one-dimensional photonic crystals and spatial dispersion in nonperiodic thin-film stacks.
We derive that, in all-pass (unity amplitude transmittance or reflectance) one-dimensional layered structures, the tangent of the group-propagation angle, the group delay, and the stored energy are approximately proportional. Thus photonic crystal superprisms and spatial dispersion in nonperiodic photonic nanostructures are generally related to wavelength-dependent stored energy in the stack.