Immunoassays with time-resolved fluorescence spectroscopy: principles and applications.
This review deals with the application of time-resolved fluorescence spectroscopy in the field of immunoassay. The advantages of time-resolved fluorescence over conventional fluorometry are discussed and it is explained why the fluorescent lanthanide chelates, and especially the Eu3+ chelates, are the labels of choice in such applications. A brief description of commercially available time-resolved fluorometers is given. The principles of phase-resolved fluorometry are described. The relative merits and disadvantages of the two time-resolved assay principles currently available commercially are critically discussed. General assay designs by using either Eu3+ or a new Eu3+ chelate as the label are also discussed in some detail. A personal view of future trends in the field of immunoassay concludes this review.


Haemodynamic, neurohumoral and exercise effects of losartan vs. captopril in chronic heart failure: results of an ELITE trial substudy. Evaluation of Losartan in the Elderly.
The AT1 receptor antagonists differ from the angiotensin converting enzyme inhibitors by achieving a more complete blockade of angiotensin II's actions and by not affecting bradykinin metabolism. There is little information on whether this causes clinically significant differences in haemodynamics, neurohormones and exercise tolerance in heart failure. To compare the effects of losartan and captopril upon central and regional haemodynamics, neurohormones and exercise capacity in heart failure. In a double-blind, randomised trial 18 patients aged > or =65 years with symptomatic heart failure were allocated to treatment with losartan (10 patients) or captopril (eight patients). Patients underwent assessment at baseline, after the first dose, at 12 weeks and at 24 weeks. Systolic blood pressure fell by - 10.7% 1 h after captopril 6.25 mg (P = 0.007) and by - 4.8% 3 h after losartan 12.5 mg (P = 0.02). The blood pressure reduction was sustained with losartan at 12 and 24 weeks. Systemic vascular resistance fell acutely after captopril (-16.4%, P = 0.01). Captopril caused an acute and sustained rise in superior mesenteric artery blood flow (+ 22.9%, P = 0.04), and a slower rise in renal artery blood flow (+31.7%, P = 0.01). Losartan had no acute effects on regional haemodynamics but had increased superior mesenteric artery blood flow by 38.1% at 12 weeks (P = 0.02). There were no substantial differences between losartan and captopril, and no changes occurred in neurohormones or exercise capacity. No substantial differences were observed between losartan and captopril on central or regional haemodynamics, neurohormones or exercise capacity in elderly patients with stable symptomatic heart failure.


Postoperative electrophysiological studies with a modified radiofrequency system. Technical aspects and clinical usefulness.
Two patients who underwent a ventricular aneurysmectomy for treatment of ventricular tachycardia are presented. In both patients, a radiofrequency pacemaker was implanted at surgery (for therapeutic use if surgery should fail). In both patients, electrophysiological studies were performed before discharge utilizing a radiofrequency pacemaker without recourse to repeat catheterization. This was possible by modifying the transmitter and coupling it to a commercially available programmable stimulator.


The distribution of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 genes of Helicobacter pylori in China.
The plasticity region of Helicobacter pylori (H. pylori) is a large chromosomal segment containing strain-specific genes. The prevalence of the plasticity region genes of the H. pylori strains in China remains unknown. The aim of this study was to examine the status of these genes and to assess the relationship between the genes and the diseases caused by H. pylori infection. A total of 141 strains were isolated from patients with chronic active gastritis (CAG), peptic ulcer disease (PUD) and gastric carcinoma (GC). The prevalence of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 was determined using PCR, and the results were analyzed using the chi-squared test. The prevalence rates of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 in the H. pylori strains were 42.55, 51.06, 20.57, 56.03 and 63.12%, respectively. The prevalence rates of jhp0940 were similar in the isolates from the CAG, PUD and GC patients, and there was no association between the jhp0940 status and any of the diseases. In contrast, the prevalence rates of jhp0945, jhp0947, jhp0949 and jhp0951 were significantly higher in the PUD and GC isolates than in the CAG isolates (p < 0.01). A univariate analysis showed that jhp0945, jhp0947, jhp0949 and jhp0951 increased the risk of PUD, while only jhp0951 was significantly associated with PUD in the multivariate analysis (p = 0.0149). The jhp0945-positive isolates were significantly associated with an increased risk for GC (p = 0.0097). The plasticity region genes are widely distributed in Chinese patients, and a high prevalence of these genes occurs in more serious diseases. Therefore, jhp0951 status is an independent factor associated with the development of PUD, and jhp0945 may predict the future development of GC in patients with CAG and is considered to be the best candidate disease marker for H. pylori-related diseases.


Acceptable symbiont cell size differs among cnidarian species and may limit symbiont diversity.
Reef-building corals form symbiotic relationships with dinoflagellates of the genus Symbiodinium. Symbiodinium are genetically and physiologically diverse, and corals may be able to adapt to different environments by altering their dominant Symbiodinium phylotype. Notably, each coral species associates only with specific Symbiodinium phylotypes, and consequently the diversity of symbionts available to the host is limited by the species specificity. Currently, it is widely presumed that species specificity is determined by the combination of cell-surface molecules on the host and symbiont. Here we show experimental evidence supporting a new model to explain at least part of the specificity in coral-Symbiodinium symbiosis. Using the laboratory model Aiptasia-Symbiodinium system, we found that symbiont infectivity is related to cell size; larger Symbiodinium phylotypes are less likely to establish a symbiotic relationship with the host Aiptasia. This size dependency is further supported by experiments where symbionts were replaced by artificial fluorescent microspheres. Finally, experiments using two different coral species demonstrate that our size-dependent-infection model can be expanded to coral-Symbiodinium symbiosis, with the acceptability of large-sized Symbiodinium phylotypes differing between two coral species. Thus the selectivity of the host for symbiont cell size can affect the diversity of symbionts in corals.


Drug-DNA intercalation: from discovery to the molecular mechanism.
The ability of small molecules to perturb the natural structure and dynamics of nucleic acids is intriguing and has potential applications in cancer therapeutics. Intercalation is a special binding mode where the planar aromatic moiety of a small molecule is inserted between a pair of base pairs, causing structural changes in the DNA and leading to its functional arrest. Enormous progress has been made to understand the nature of the intercalation process since its idealistic conception five decades ago. However, the biological functions were detected even earlier. In this review, we focus mainly on the acridine and anthracycline types of drugs and provide a brief overview of the development in the field through various experimental methods that led to our present understanding of the subject. Subsequently, we discuss the molecular mechanism of the intercalation process, free-energy landscapes, and kinetics that was revealed recently through detailed and rigorous computational studies.


Aneurysmal bone cyst of the sacrum.
A series of five cases of sacral aneurysmal bone cyst is presented. There were three females and two males, the youngest patient being 11 years old, the oldest 41 years old. All patients had pain of long duration with varying signs of nerve root compression. Three patients had micturitional problems. Four patients had a palpable mass which was tender. All had standard radiographs. Tomograms, computed tomography (CT) scans or MRI (magnetic resonance imaging) were obtained in some patients. In all patients a definite histologic diagnosis was established. Four lesions healed after resection (two cases) or curettage (two cases). One patient is considered to have a stable lesion which is not progressing.


Effects of calmodulin on aromatase activity in the preoptic area.
Oestrogens derived from the neural aromatisation of testosterone play a key role in the activation of male sexual behaviour in many vertebrates. Besides their slow action on gene transcription mediated by the binding to nuclear receptors, oestrogens have now been recognised to have more rapid membrane-based effects on brain function. Rapid changes in aromatase activity, and hence in local oestrogen concentrations, could thus rapidly modulate behavioural responses. We previously demonstrated that calcium-dependent kinases are able to down-regulate aromatase activity after incubations of 10-15 min in phosphorylating conditions. In the present study, in quail hypothalamic homogenates, we show that Ca2+ or calmodulin alone can very rapidly change aromatase activity. Preincubation with 1 mM EGTA or with a monoclonal antibody raised against calmodulin immediately increased aromatase activity. The presence of calmodulin on aromatase purified by immunoprecipitation and electrophoresis was previously identified by western blot and two consensus binding sites for Ca2+-calmodulin are identified here on the deduced amino acid sequence of the quail brain aromatase. The rapid control of brain aromatase activity thus appears to include two mechanisms: (i) an immediate regulatory process that involves the Ca2+-calmodulin binding site and (ii) a somewhat slower phosphorylation by several protein kinases (PKC, PKA but also possibly Ca2+-calmodulin kinases) of the aromatase molecule.


[Behaviour of the prolyl hydroxylase to collagen synthesis in KB cells].
KB cells, which synthetized collagen at a low rate, shown a prolyl hydroxylase activity at the same rate that fibroblast. The relationship between collagen synthesis and prolyl hydroxylase activity in these cells was discussed.


Co-creating a Peer Education program to improve skin health in older people from diverse communities: An innovation in health promotion.
Chronic wounds, debilitating and costly to manage, are more common in older people. Prevention is possible through improving skin health. We developed, implemented and evaluated an innovative health promotion program to improve skin health of older adults. A one-hour, peer education program was co-created and delivered to culturally diverse community-dwelling older people. A mixed-methods evaluation approach comprised objective measures of skin health and barrier function at commencement and six weeks posteducation, and focus groups posteducation. Seventy-three participants participated in the study (mean age 74.38 ± 11.80 years). Hydration significantly improved at follow-up for English speaking participants (t(27) = -2.90, P = 0.007). The majority of participants reported the education to be informative and useful in supporting behaviour changes. The peer education program improved skin hydration in older English speaking individuals. Peer education may effectively deliver health promotion information in some groups.


Validation and cross-cultural adaptation of the National Eye Institute Visual Function Questionnaire (NEI VFQ-25) in Serbian patients.
To test the validity and reliability of the Serbian version of the interviewer-administered format of the National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25). The Serbian version of NEI VFQ-25 was translated in accordance with standard methods that have been adopted internationally. In order to assess the reliability and validity of the translated NEI VFQ-25, we used a sample of 105 patients with four different chronic ocular diseases. Cronbach's alpha coefficient was used to assess internal consistency for each subscale. To assess test-retest reliability, intraclass correlation coefficients were used. The test-retest data were obtained from clinically stable patients with age-related cataracts, in surveys performed 2 weeks apart. Rasch analysis was also applied as a modern methods of psychometric assessment of the questionnaire. Four groups of patients were studied and the most prevalent were patients with cataract 40 (38.1%), followed by diabetic retinopathy 31 (29.5%), age related macular degeneration 22 (21.0%) and glaucoma 12 (11.4%). The overall index score on the NEI VFQ-25 ranged from 65.3 to 67.8 with a mean of 67.4 ± 15.0. Cronbach's alpha coefficient (index of internal consistency reliability) ranged from 0.643 to 0.889 for the subscales. Evaluation of the validity of the Serbian version of NEI VFQ-25 is presented in the multi-trait-multi-method matrix and all items passed the convergent and discriminant validity tests. Rasch analysis showed a good measurement precision, but also demonstrated misfitting items and multidimensionality of the questionnaire. Although traditional validation method indicates that the Serbian version of NEI VFQ-25 is a valid and reliable instrument for the assessment of vision specific QoL in Serbian populations aged 40 years or older, Rasch analysis revealed a substantial weakness of the questionnaire that should be taken into consideration when interpreting the results.


Carcinoma of the colon in the adolescent: a report of survival and an analysis of the literature.
A case of adenocarcinoma of the transverse colon in a fourteen year old girl is presented. Five years after radical resection of this Dukes' C2 lesion, the patient is alive and well without recurrent disease. The incidence of large bowel carcinoma in adolescents remains low, but there is an increasing tendency toward neoplasia in younger patients. Factors that affect patient survival included late stage at presentation, delay in treatment, paucity of symptoms, and a higher incidence of tumors of the transverse colon. An increased incidence of colloid carcinoma is noted. The etiology and pathogenesis of large bowel cancer as it pertains to childhood are reviewed.


Evaluation of strategies to improve the production of alkaline protease PrtA from Aspergillus nidulans.
Aspergillus nidulans produces several proteases. The prtA gene encodes a major protease, and two approaches were explored to achieve the overproduction of this enzyme. Molecular cloning of the mature form of this enzyme in Pichia pastoris resulted in the production of an inactive form. In addition, the presence of this enzyme was toxic for the host and resulted in cell lysis. The modification of the culture medium constituents resulted in a 6.4-fold increase in enzyme production. The main effect was achieved through the use of organic nitrogen sources. Although it was previously shown that the PrtA protease shows promiscuous esterase activity, the production of this enzyme was not induced by lipidic sources.


[Efficacy and safety of Fufang Biejia Ruangan tablet in patients with chronic hepatitis B complicated with hepatic fibrosis].
To study the clinical therapeutic effects and safety of Fufang Biejia Ruangan tablet (FBRt) in patients with chronic hepatitis B complicated with hepatic fibrosis. Totally 420 patients were randomly divided into two groups, FBRt group (300 cases) were treated with Fufang Biejia Ruangan tablets and control group (120 cases) were treated with He Luo Shu Gan capsule, the patients in both groups were treated for 6 months. The cure rate and total effective rate of FBRt group were significantly higher than those of control group (55.67 percent and 81.67 percent vs. 15.8 percent and 60.00 percent, P less than 0.01). Fufang Biejia Ruangan tablet could alleviate clinical symptoms and hepatic fibrosis. Fufang Biejia Ruangan tablet is effective and safe in treatment of patients with chronic hepatitis B complicated with liver fibrosis.


fNIRS evidence of prefrontal regulation of frustration in early childhood.
The experience of frustration is common in early childhood, yet some children seem to possess a lower tolerance for frustration than others. Characterizing the biological mechanisms underlying a wide range of frustration tolerance observed in early childhood may inform maladaptive behavior and psychopathology that is associated with this construct. The goal of this study was to measure prefrontal correlates of frustration in 3-5-year-old children, who are not readily adaptable for typical neuroimaging approaches, using functional near infrared spectroscopy (fNIRS). fNIRS of frontal regions were measured as frustration was induced in children through a computer game where a desired and expected prize was "stolen" by an animated dog. A fNIRS general linear model (GLM) was used to quantify the correlation of brain regions with the task and identify areas that were statistically different between the winning and frustrating test conditions. A second-level voxel-based ANOVA analysis was then used to correlate the amplitude of each individual's brain activation with measure of parent-reported frustration. Experimental results indicated increased activity in the middle prefrontal cortex during winning of a desired prize, while lateral prefrontal cortex activity increased during frustration. Further, activity increase in lateral prefrontal cortex during frustration correlated positively with parent-reported frustration tolerance. These findings point to the role of the lateral prefrontal cortex as a potential region supporting the regulation of emotion during frustration.


Evaluation, grading, and use of the RIME vocabulary on internal medicine clerkships: results of a national survey and comparison to other clinical clerkships.
Evaluation methods within and across clerkships are rapidly evolving, including greater emphasis or frameworks for descriptive evaluation and direct observation of competence. The purpose of this study is to describe current evaluation methods, use of the Reporter-Interpreter-Manager/Educator (RIME) framework, and grade assignment by internal medicine clerkship directors. In 2005, the Clerkship Directors in Internal Medicine surveyed its 109 institutional members. Topics included evaluation methods and grade contribution, use of evaluation sessions and/or RIME, and grade assignment (criterion referenced or normative). Response rate was 81% (88/109). The evaluation methods were as follows: teachers' evaluations, 93% (64% of grade); National Board of Medical Examiners subject examination, 81% (25% of grade); faculty written exam, 34% (14% of grade); objective structured clinical examinations, 32% (12% of grade); direct observation, 22% (7% of grade). RIME is used by 42% of respondents. Many clerkship directors (43%) meet with teachers to discuss student performance. Criterion-referenced grading is used by 59%, and normative grading is used by 27%. Unsatisfactory grades are given for examination failures (72%), unprofessional behavior (49%), poor clinical performance (42%), and failure to meet requirements (18%). Internal medicine clerkship directors emphasize description and observation of students. RIME and discussions with teachers are becoming commonplace.


Surface (S)-layer proteins of Deinococcus radiodurans and their utility as vehicles for surface localization of functional proteins.
The radiation resistant bacterium, Deinococcus radiodurans contains two major surface (S)-layer proteins, Hpi and SlpA. The Hpi protein was shown to (a) undergo specific in vivo cleavage, and (b) closely associate with the SlpA protein. Using a non-specific acid phosphatase from Salmonella enterica serovar Typhi, PhoN as a reporter, the Surface Layer Homology (SLH) domain of SlpA was shown to bind deinococcal peptidoglycan-containing cell wall sacculi. The association of SlpA with Hpi on one side and peptidoglycan on the other, localizes this protein in the 'interstitial' layer of the deinoccocal cell wall. Gene chimeras of hpi-phoN and slh-phoN were constructed to test efficacy of S-layer proteins, as vehicles for cell surface localization in D. radiodurans. The Hpi-PhoN protein localized exclusively in the membrane fraction, and displayed cell-based phosphatase activity in vivo. The SLH-PhoN, which localized to both cytosolic and membrane fractions, displayed in vitro activity but no cell-based in vivo activity. Hpi, therefore, emerged as an efficient surface localizing protein and can be exploited for suitable applications of this superbug.


Prey and predator emigration responses in the acarine system Tetranychus urticae-Phytoseiulus persimilis.
Some of the processes that influence the emigration of prey and predatory mites from bean plants were investigated experimentally. The emigration of the prey depends on the damage they cause to the plants and on predator density. The predator's emigration rate is a decreasing function of prey density, and does not change (or it slightly decreases) when prey and predator numbers are increased maintaining the same prey/predator ratio. The probability of emigration of the predators is independent of their own density when prey are absent and density dependent when prey density is kep constant. Forty three per cent of the variability in the predator's instantaneous rate of emigration in the different experiments is accounted for by a two parameter negative exponential function of capture rate (number of prey eaten per predator and per unit of time).


Fatality secondary to misuse of TAC solution.
The death of a 7 1/2-month-old girl from the misuse of tetracaine/adrenalin/cocaine solution for wound anesthesia is reported. Ten milliliters of the solution inappropriately came into contact with nasal mucous membranes, causing excessive drug absorption. The patient's death probably was due to cocaine toxicity (post-mortem blood level, 11.9 mg/L).


The Shaw haemostatic scalpel in paediatric surgery: clinical report on 3000 operations.
This report presents the experience in a prospective series of 100 operative procedures in all fields of paediatric surgery with the Shaw haemostatic scalpel, which we have subsequently used in more than 3000 further operations. The Shaw scalpel proved to be advantageous in about 80% of major cases. The scalpel cuts tissue with a sharp steel edge, like a cold scalpel, and simultaneously seals blood vessels by heat thermally conducted to the tissue from heated blade which is electrically insulated from the patient. The heat seals most small blood vessels (under 2 mm) as they are cut. Since no electric current passes through the patient, a grounding pad is not needed and the risk of accidental electrical current burns at grounding sites is eliminated. Muscle stimulation associated with the use of a normal cautery is avoided, improving surgical precision of cutting. The Shaw haemostatic scalpel minimizes damage to the tissue as compared with other thermocoagulating instruments. Since it seals small vessels as it cuts tissue, it largely eliminates the flow of blood into the incised area and allows better visibility of the surgical field. The use of the scalpel requires a different cutting technique which is however easy to learn. The Shaw haemostatic scalpel reduces blood loss and overall operating time in major cases. It is relatively inexpensive and can be recommended for use in paediatric surgery.


Still no single gold standard for using cementless femoral stems routinely in total hip arthroplasty.
There is now a clear trend with increased usage of cementless femoral stems for all ages and most patients. As the number of total hip arthroplasties (THAs) performed annually continues to increase with expanding indications for THA and demands for improved quality of life, so will the prevalence of THA in the elderly and aging populations. This is worrisome as the risk of complications with cementless femoral stems increases in elderly patients and those with poor bone quality. The purpose of this study is to analyze the available data from comparative studies to determine whether cementless femoral stems are overused and whether cemented stems warrant increased consideration. Using Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, searches were performed to find original studies comparing cementless and cemented femoral stems; large population registry studies and reports were also included. This systematic review documents that older patients with cementless fixation increase the risk of revision, there is no clear fixation advantage in midaged patients, and younger patients fare better with cementless fixation. Complications after THA create burdens on the health care system and on patients. Using evidence-based data should be better guidance in selecting the most reliable implants for THA. Although cementless femoral fixation for THA has evolved to the "new standard," it has not been proven to be the "gold standard" for all patients.


QSAR workbench: automating QSAR modeling to drive compound design.
We describe the QSAR Workbench, a system for the building and analysis of QSAR models. The system is built around the Pipeline Pilot workflow tool and provides access to a variety of model building algorithms for both continuous and categorical data. Traditionally models are built on a one by one basis and fully exploring the model space of algorithms and descriptor subsets is a time consuming basis. The QSAR Workbench provides a framework to allow for multiple models to be built over a number of modeling algorithms, descriptor combinations and data splits (training and test sets). Methods to analyze and compare models are provided, enabling the user to select the most appropriate model. The Workbench provides a consistent set of routines for data preparation and chemistry normalization that are also applied for predictions. The Workbench provides a large degree of automation with the ability to publish preconfigured model building workflows for a variety of problem domains, whilst providing experienced users full access to the underlying parameterization if required. Methods are provided to allow for publication of selected models as web services, thus providing integration with the chemistry desktop. We describe the design and implementation of the QSAR Workbench and demonstrate its utility through application to two public domain datasets.


The mechanism of liver microsomal lipid peroxidation.
In the presence of Fe-3+ and complexing anions, the peroxidation of unsaturated liver microsomal lipid in both intact microsomes and in a model system containing extracted microsomal lipid can be promoted by either NADPH and NADPH : cytochrome c reductase or by xanthine and xanthine oxidase. Erythrocuprein effectively inhibits the activity promoted by xanthine and xanthine oxidase but produces much less inhibition of NADPH-dependent peroxidation. The singlet-oxygen trapping agent, 1, 3-diphenylisobenzofuran, had no effect on NADPH-dependent peroxidation but strongly inhibited the peroxidation promoted by xanthine and xanthine oxidase. NADPH-dependent lipid peroxidation was also shown to be unaffected by hydroxyl radical scavengers.. The addition of catalase had no effect on NADPH-dependent lipid peroxidation, but it significantly increased the rate of malondialdehyde formation in the reaction promoted by xanthine and xanthine oxidase. The results demonstrate that NADPH-dependent lipid peroxidation is promoted by a reaction mechanism which does not involve either superoxide, singlet oxygen, HOOH, or the hydroxyl radical. It is concluded that NADPH-dependent lipid peroxidation is initiated by the reduction of Fe-3+ followed by the decomposition of hydroperoxides to generate alkoxyl radicals. The initiation reaction may involve some form of the perferryl ion or other metal ion species generated during oxidation of Fe-2+ by oxygen.


To what extent do citizens support the disinvestment of healthcare interventions? An exploration of the support for four viewpoints on active disinvestment in the Netherlands.
Active disinvestment of healthcare interventions (i.e. discontinuing reimbursement by means of a policy decision) has received limited public support in the past. Previous research has identified four viewpoints on active disinvestment among citizens in the Netherlands. However, it remained unclear how strong these viewpoints are supported by society, and by whom. Therefore, the current study aimed to 1) measure the support for these four viewpoints and 2) assess whether support is associated with background characteristics of citizens. In an online survey, a representative sample of adult citizens in the Netherlands (n = 1794) was asked to rate their agreement with short narratives of the four viewpoints on a 7-point Likert scale. The survey also included questions on sociodemographic characteristics, health status, healthcare utilization, and opinions about responsibility and costs in the healthcare context. Logistic regression models were estimated for each viewpoint to assess the association between viewpoint support and these characteristics. The support for the different viewpoints varied between 46.8% and 57.7% of the sample. Viewpoint support was associated with participants' age, gender, educational level, financial situation, healthcare utilization, opinion on the responsibility of the government for the health of citizens, and opinion on whether the increase in healthcare expenditure and health insurance premiums is considered a problem. Resistance to active disinvestment may partially be explained by the consequences of disinvestment citizens anticipate experiencing themselves. Citizens considering the increase in healthcare expenditure a larger problem were more supportive of disinvestment than those considering it less of a problem.


Comparison of different tests assessing neurosensory disturbances after bilateral sagittal split osteotomy.
Different methods of assessing functional impairment of the inferior alveolar nerve after sagittal split osteotomy were compared with subjective reports. Thirty patients were tested preoperatively as well as after four days, three weeks, three months, six months, and one year of follow up. All the patients underwent bilateral mandibular ramus sagittal split osteotomy. The sensation of the mandibular teeth was assessed by an electric vitality scanner. Other neurosensory assessments included light touch, two-point discrimination, tactile discrimination, and thermal discrimination. It was shown that electric sensibility-testing of mandibular teeth is a useful method for evaluating neurosensory disturbance after sagittal split osteotomy, and it correlated even better with the patient's subjective evaluation than the other tests used in this study.


Azathioprine-Induced Marrow Suppression in Dermatology Patients - Analysis of 18 Patients.
Azathioprine is an immunosuppressant used to treat several immunological disorders. As a purine analog, it inhibits DNA synthesis and cell multiplication. However, marrow suppression is a serious complication associated with azathioprine. To analyze the marrow suppression caused by azathioprine in dermatology patients. This is a retrospective analysis of the records of 18 patients who presented with marrow suppression secondary to azathioprine which was used for the treatment of various dermatological diseases. The analysis includes 18 patients, 15 females and 3 males with the average age being 25.88 years. All except two patients received 1 mg/kg of oral azathioprine once daily. Leukopenia was seen in 13 patients (with severe leukopenia in 7 patients), thrombocytopenia in 8, and low hemoglobin in 14 patients. Isolated low hemoglobin was seen in four patients, isolated leukopenia in four patients, and only one patient presented with isolated thrombocytopenia. Six patients had pancytopenia. The duration from the starting dose to reporting of marrow suppression ranged from 10 days to 1 year. Eight out of 18 patients presented with anagen effluvium, 2 patients with oral ulcers, and 1 patient with an upper respiratory tract infection. All the patients recovered within 1 month. Marrow suppression due to azathioprine can occur with a low dose of 1 mg/kg. Hair loss and oral ulcers serve as early warning signs for marrow suppression.


Serum FLT-3 ligand in a busulphan-induced model of chronic bone marrow hypoplasia in the female CD-1 mouse.
The concentration of the cytokine fms-like tyrosine kinase-3 ligand (FL) is elevated in the plasma of patients treated with chemotherapy or radiotherapy for malignant conditions. In addition, plasma FL is increased in patients with bone marrow failure resulting from stem-cell defects (e.g. aplastic anaemia). Our goal in the present study was to measure the concentration of serum FL in mice treated with the chemotherapeutic agent busulphan (BU) to induce bone marrow depression and relate changes in FL to effects on haemopoiesis. Female CD-1 mice were treated with BU (9.0 mg/kg) or vehicle by intraperitoneal injection on 10 occasions over 21 days. Animals were autopsied on days 1, 23, 72, 119 and 177 postdosing. A full blood count was performed, and serum prepared for FL analysis. Femoral marrow cell suspensions were prepared to assess the total femoral nucleated cell count (FNCC) and the number of committed haemopoietic progenitor cells (CFU-C). On days 1 and 23 postdosing, significant decreases were evident in many peripheral blood parameters; the FNCC and CFU-C were also reduced in BU-treated mice, in conjunction with increases in serum FL levels. On days 72, 119 and 177 postdosing, several peripheral blood and bone marrow parameters remained reduced and the concentration of serum FL continued to be significantly increased. Linear regression analysis demonstrated significant correlations between the concentration of serum FL in BU-treated mice and peripheral blood and bone marrow parameters; this suggests the possible use of serum FL as a potential biomarker for drug-induced bone marrow injury.


Gonococcal endocarditis after a threatened abortion. A case report.
Disseminated gonocococcal infection arising during pregnancy is an uncommon occurrence. Gonococcemia following a threatened abortion has not been documented previously. Gonococcal arthritis and endocarditis in a 22-year-old primigravida occurred following a midtrimester threatened abortion. Since gonococcemia is thought to be secondary to endocervical disruption, chemoprophylaxis in the gravid cardiac patient after a threatened abortion is recommended.


Embryonic heart rate in the early first trimester: what rate is normal?
Embryonic heart rates below 90 beats per minute at 6 to 8 weeks of gestation have been shown to be associated with a high likelihood of subsequent first trimester demise. The boundary between slow and normal heart rates has not been established, however, in that previous studies do not identify the heart rate for each gestational age at which prognosis plateaus. We evaluated sonograms of singleton pregnancies at 6 to 8 weeks to determine the relationship between heart rate and first trimester outcome in four gestational age subgroups: < or = 6.2, 6.3-7.0, 7.1-7.5, and 7.6-8.0 weeks. Below 6.2 weeks, prognosis improved as heart rate increased to 100 beats per minute, then plateaued for rates > or = 100 beats per minute. At 6.3-7.0 weeks, prognosis improved up to 120 beats per minute, then plateaued. Heart rate norms could not be established at 7.1-8.0 weeks because of the small number of embryonic or fetal deaths in this age group; however, all embryos with heart rates below 110 beats per minute at 7.1-8.0 weeks died. We conclude that the lower limit of normal is 100 beats per minute up to 6.2 weeks' gestation and 120 bpm at 6.3-7.0 weeks.


Influenza virus-model membrane interaction. A morphological approach using modern cryotechniques.
The membrane fusion activity of influenza virus was characterized morphologically using a model system composed of a highly purified influenza B virus suspension and ganglioside-containing zwitterionic liposomes. Electron microscopical analysis was performed after a combination of fast-freezing with either freeze-fracture or freeze-substitution-thin sectioning, ensuring maximal time resolution and avoiding preparation artifacts. In a parallel fluorescence 'lipid mixing' fusion assay, influenza virus-membrane fusion was characterized biochemically. Biochemical and morphological data are in full agreement, indicating negligible membrane fusion activity at neutral pH and high fusion activity at low pH. The freeze-fracture morphology strongly suggests a local point contact between viral and liposomal membrane at neutral pH, and a local point fusion mechanism for influenza virus-membrane fusion upon lowering of the pH. Fusion is followed by lipid mixing, lateral diffusion of viral spike proteins and exposure of viral contents at the inner liposomal surface.


Estimating the Within-Person Change in Dental Service Access Measures during the COVID-19 Pandemic in the United States.
American adults delay dental care more than any other healthcare service. Unfortunately, the COVID-19 pandemic may have stalled efforts to address dental service delays. Early evidence has suggested substantial declines in dental service visits in the early phase of the pandemic; however, our study is among the first to measure within-person changes from 2019 to 2020 and conduct subgroup analyses to examine if changing dental patterns were mediated by exposure to the pandemic, risk of adverse COVID-19 outcomes, or dental insurance. We analyzed a National Health Interview Survey panel of individuals initially surveyed in 2019, with subsequent follow-up in 2020. The outcomes included dental service access measures and the interval of a most recent dental visit. By constructing a probability-weighted linear regression model with fixed-effects, we estimated the average within-person change from 2019 to 2020. Robust standard errors were clustered within each respondent. From 2019 to 2020, adults reported a 4.6%-point reduction in the probability of visiting the dentist (p < 0.001). Significantly higher declines were found in Northeast and West regions compared to Midwest and South regions. We find no evidence that declining dental services in 2020 were associated with more chronic diseases, older age, or lack of dental insurance coverage. Adults did not report more financial or nonfinancial access barriers to dental care in 2020 compared to 2019. The long-term effects of the COVID-19 pandemic on delayed dental care warrant continued monitoring as policymakers aim to mitigate the pandemic's negative consequences on oral health equity.


Direct PCR detection of Escherichia coli O157:H7.
This paper reports a simple, rapid approach for the detection of Shiga toxin (Stx)-producing Escherichia coli (STEC). Direct PCR (DPCR) obviates the need for the recovery of cells from the sample or DNA extraction prior to PCR. Primers specific for Stx-encoding genes stx1 and stx2 were used in DPCR for the detection of E. coli O157:H7 added to environmental water samples and milk. PCR reactions containing one cell yielded a DPCR product. This should provide an improved method to assess contamination of environmental and other samples by STEC and other pathogens.


Effect of PON1 Q192R genetic polymorphism on clopidogrel efficacy and cardiovascular events in the Clopidogrel in the Unstable Angina to Prevent Recurrent Events trial and the Atrial Fibrillation Clopidogrel Trial with Irbesartan for Prevention of Vascular Events.
A recent report suggested that carriers of the Q allele of the PON1 Q192R polymorphism had decreased biotransformation of clopidogrel into its active metabolite and decreased efficacy of clopidogrel in preventing cardiovascular events. Furthermore, PON1 has been reported to have a central role in the antioxidant function of high-density lipoprotein, and the Q192R polymorphism has been previously associated with cardiovascular risk in patients not treated with clopidogrel. Patients (n=5059) from the Clopidogrel in Unstable Angina to Prevent Recurrent Events (CURE) randomized trial that demonstrated benefits of clopidogrel versus placebo in preventing cardiovascular events in acute coronary syndromes were genotyped for the PON1 Q192R polymorphism. Clopidogrel compared with placebo significantly reduced the first primary efficacy outcome, irrespective of PON1 Q192R genotype (P=0.07 for heterogeneity). No association was observed between the Q192R polymorphism and cardiovascular events in the overall sample (hazard ratio [HR], 1.09 per allele; 95% confidence interval [CI], 0.95-1.24; P=0.23). However, an association was observed between the Q allele and increased cardiovascular events in the placebo group (HR, 1.23 per allele; 95% CI, 1.03-1.47; P=0.03) but not in the clopidogrel group (HR, 0.93 per allele; 95% CI, 0.76-1.13; P=0.46). In 1156 atrial fibrillation patients from the Atrial Fibrillation Clopidogrel Trial With Irbesartan for Prevention of Vascular Events, there was no evidence of interaction between PON1 genotype and clopidogrel for any outcome or for an association between genotype and cardiovascular events. In conclusion, our study shows that PON1 Q192R genotype does not modify the efficacy and safety of clopidogrel in patients with acute coronary syndromes. Further studies are needed to confirm or refute the association of the Q allele with adverse cardiovascular events independent of clopidogrel in secondary prevention patients.


Panic disorder respiratory subtype: a comparison between responses to hyperventilation and CO2 challenge tests.
In this study 117 panic disorder patients were divided into a respiratory subtype group and a non-respiratory subtype group. The respiratory subtype patients were observed to be more sensitive to the 35% CO(2) inhalation challenge test and the hyperventilation test than the non-respiratory subtype patients.


Characterization of an antibody to the cross-reacting determinant of the glycosyl-phosphatidylinositol anchor of human membrane dipeptidase.
A polyclonal antiserum raised to the phospholipase C-solubilized form of membrane dipeptidase (EC 188.8.131.52) purified from human kidney was found to cross-react with unrelated trypanosomal and porcine glycosyl-phosphatidylinositol anchored proteins. Those antibodies recognising the cross-reacting determinant (CRD) were isolated by chromatography on a column of immobilized phospholipase C-solubilized porcine aminopeptidase P (EC 220.127.116.11), and the epitopes involved in the recognition were then characterized by immunoelectrophoretic blot analysis and by a competitive ELISA. The phospholipase C-solubilized forms of human and porcine membrane dipeptidase, porcine aminopeptidase P and trypanosome variant surface glycoprotein were recognised by the anti-CRD antiserum, and this recognition was abolished by prior treatment of the proteins with either mild acid or nitrous acid. In contrast, the detergent-solubilized, membrane-forms of human and porcine membrane dipeptidase were not recognised. Of a range of components of the glycosyl-phosphatidylinositol anchor, only inositol 1,2-cyclic monophosphate and the insulin-mimetic disaccharide, glucosaminyl-1,6-inositol 1,2-cyclic monophosphate, inhibited in the micromolar range the binding of the anti-CRD antiserum to immobilized porcine aminopeptidase P. These results indicate that the major epitope recognised by this anti-CRD antiserum is the inositol 1,2-cyclic monophosphate formed on phospholipase C cleavage of the glycosyl-phosphatidylinositol anchor.


PM1-loaded recombinant human H-ferritin nanocages: A novel pH-responsive sensing platform for the identification of cancer cells.
The aggregation-induced emission (AIE) material has been widely used in biological detection due to their unique property of fluorescing in aggregation state. However, the poor dispersion and biocompatibility limit its application in in vivo real-time imaging. Here, a novel strategy is designed to obtain pH-responsive AIE nanomaterials, working through 4-Undecoxy Tetraphenyl Ethylene Methacrylate (PM1) block, with excellent features (dispersion, biocompatibility, self-reconstruction and cancer specific recognition). The recombinant human H-ferritin (rHuHF) was used to prepare rHuHF-PM1 nanocomposites which effectively supported the dispersion and transfer of PM1 in the biological environment, even making it target tumor cells due to the overexpression of ferritin receptors on tumor cells. To simulate the changes of rHuHF in intracellular lysosomes, particle size and fluorescence of rHuHF-PM1 were analyzed, which reflected the loose structural changes of rHuHF nanocages in weak acid system that facilitated the degradation of macromolecular rHuHF in intracellular lysosomes and following release of PM1. The released PM1 molecules aggregated and emitted brilliant blue fluorescence. Several cell lines, Hela, HT-29, HepG2, L-O2 and HUVEC have all been sensitively detected and distinguished. Accordingly, this nanocage has a potential to be applied to disease diagnosis and provides a novel sensing platform for the identification of cancer.


Kinase inhibitor profiling using chemoproteomics.
Quantitative chemoproteomics has recently emerged as an experimental approach to determine protein interaction profiles of small molecules in a given cell line or tissue. In contrast to standard biochemical and biophysical kinase assays, application of this method to kinase inhibitors determines compound binding to endogenously expressed kinases under conditions approximating the physiological situation with regard to the molecular state of the kinase and presence of required cofactors and regulatory proteins. Using a dose-dependent, competition-based experimental design in combination with quantitative mass spectrometry approaches, such as the use of tandem mass tags (TMT) for isobaric labeling described here, allows to rank-order interactions of inhibitors to kinase by binding affinity.


Effect of rectal diclofenac and acetaminophen alone and in combination on postoperative pain after cleft palate repair in children.
Acetaminophen and diclofenac are prescribed as postoperative analgesic agents in children. However, the efficacy of their combination is not studied sufficiently. We compare the analgesic effects of acetaminophen, diclofenac, and their combination after cleft palate surgery. In this randomized clinical trial, 120 children (1.5-5 y) who were scheduled for cleft palate repair were studied. Children were randomized to receive placebo, acetaminophen (40 mg/kg), diclofenac (1 mg/kg), or acetaminophen (40 mg/kg) plus diclofenac (1 mg/kg) rectally just after surgery. Acetaminophen (30 mg/kg) and diclofenac (1 mg/kg) were administered every 8 hours until 48 hours. Postoperative pain was assessed regularly with the Children Hospital of Eastern Ontario Pain Scale, and rescue analgesia was provided if scores were 7 or greater. Time to the first prescription of meperidine, total postoperative meperidine consumption, and adverse effects were the main outcomes.After surgery, pain scores were higher in placebo than in other groups in all time intervals. In the first 12 hours, pain scores in the combined group were less than those in the acetaminophen (P < 0.05) and diclofenac (P < 0.05) groups. Postoperative meperidine consumption was the highest in placebo and was the least in combined group (P < 0.05). It was significantly higher in the acetaminophen group than in the diclofenac group (P < 0.05). Time to the first prescription of meperidine was significantly different among all groups. Adverse effects were comparable among groups.Rectal acetaminophen plus diclofenac was found to be the most effective in pain control. However, both rectal acetaminophen and diclofenac were more effective than placebo, whereas diclofenac was more effective than acetaminophen.


The BDNF-Val66Met polymorphism: implications for susceptibility to multiple sclerosis and severity of disease.
Neurodegeneration following inflammatory injury is considered to be a pathological correlate of irreversible disability in patients with multiple sclerosis. The availability of neurotrophins could influence the probability or rate of disease progression and the time of onset. The BDNF-Val66Met-polymorphism leads to altered intracellular transport and secretion of BDNF, and is thus a logical candidate for a gene that influences susceptibility and, more specifically, the clinical course of multiple sclerosis. In order to test this hypothesis we genotyped the polymorphism in 951 UK multiple sclerosis trio families, but found no evidence for association before (p=0.63) or after stratification for clinical course (p=0.73).


Biochemistry and physiology of the natriuretic peptide receptor guanylyl cyclases.
Guanylyl cyclases (GC) exist as soluble and particulate, membrane-associated enzymes which catalyse the conversion of GTP to cGMP, an intracellular signalling molecule. Several membrane forms of the enzyme have been identified up to now. Some of them serve as receptors for the natriuretic peptides, a family of peptides which includes atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP), three peptides known to play important roles in renal and cardiovascular physiology. These are transmembrane proteins composed of a single transmembrane domain, a variable extracellular natriuretic peptide-binding domain, and a more conserved intracellular kinase homology domain (KHD) and catalytic domain. GC-A, the receptor for ANP and BNP, also named natriuretic peptide receptor-A or -1 (NPR-A or NPR- 1), has been studied widely. Its mode of activation by peptide ligands and mechanisms of regulation serve as prototypes for understanding the function of other particulate GC. Activation of this enzyme by its ligand is a complex process requiring oligomerization, ligand binding, KHD phosphorylation and ATP binding. Gene knockout and genetic segregation studies have provided strong evidence for the importance of GC-A in the regulation of blood pressure and heart and renal functions. GC-B is the main receptor for CNP, the latter having a more paracrine role at the vascular and venous levels. The structure and regulation of GC-B is similar to that of GC-A. This chapter reviews the structure and roles of GC-A and GC-B in blood pressure regulation and cardiac and renal pathophysiology.


Correlation of Manifest Refraction and Simulated Keratometry to Tomography Characteristics in Patients With Keratoconus.
To report on baseline refractive and keratometric values and their correlation with tomographic characteristics of eyes with keratoconus (KC). Retrospective chart review of patients treated in a single-center cornea and refractive surgery practice. Baseline topographic measurements were reviewed for 1,012 keratoconic eyes of 586 patients between 2008 and 2018. The manifest refraction, thinnest pachymetry (P thin ), corneal astigmatism (K astig ), and the maximum (K max ), steep (K steep ), flat (K flat ), and mean (K mean ) keratometry were analyzed. The location of K max (x, y) was used to determine central (<1 mm), paracentral (1-3 mm), pericentral (3-5 mm), or peripheral (>5 mm) cone locations. In the entire cohort, the mean manifest sphere was -2.2±4.4 diopters (D) and the cylinder was -3.2±2.3 D. In total, 48.6% of patients had against the rule (ATR) manifest astigmatism (M astig ). The average K astig was 3.8±2.7 D, and unlike the manifest axis, 50.2% of patients had with the rule (WTR) K astig . Patients with a K max less than 50 D had an M astig of -1.9±1.6 D, 45.9% of which was ATR M astig . With respect to baseline tomography measurements, K max , K steep , K flat , and K mean were 58.0±9.4, 50.6±6.5, 46.8±5.9, and 48.6±6.1 D, respectively. There was a weak correlation between K max and simulated keratometry (K steep , K flat , and K mean ) for patients with a K max less than 60 D. Simulated keratometry is poorly correlated with KC severity until the disease is more severe. M astig ≥2 D and ATR M astig were correlated with KC at all levels of severity. M astig ≥2 D and ATR M astig may serve as a simple, inexpensive, and widely available indicator for topographic analysis to identify possible KC and suggest further workup; however, further prospective studies are needed to confirm its utility.


An aminotransferase branch point connects purine catabolism to amino acid recycling.
Although amino acids are known precursors of purines, a pathway for the direct recycling of amino acids from purines has never been described at the molecular level. We provide NMR and crystallographic evidence that the PucG protein from Bacillus subtilis catalyzes the transamination between an unstable intermediate ((S)-ureidoglycine) and the end product of purine catabolism (glyoxylate) to yield oxalurate and glycine. This activity enables soil and gut bacteria to use the animal purine waste as a source of carbon and nitrogen. The reaction catalyzed by (S)-ureidoglycine-glyoxylate aminotransferase (UGXT) illustrates a transamination sequence in which the same substrate provides both the amino group donor and, via its spontaneous decay, the amino group acceptor. Structural comparison and mutational analysis suggest a molecular rationale for the functional divergence between UGXT and peroxisomal alanine-glyoxylate aminotransferase, a fundamental enzyme for glyoxylate detoxification in humans.


Genetic parameters for mid-infrared spectroscopy-predicted fertility.
Female fertility is a challenging trait to improve genetically because of its low heritability, its unfavorable genetic correlation with milk yield, and its relatively small number of records. The MFERT trait is the probability of conception to first insemination predicted using mid-infrared (MIR) spectroscopy of a milk sample collected during lactation as part of routine milk recording, age at calving, days in milk, and milk production. As such, MFERT could become available on many more cows than traditional fertility traits. Our objectives were (1) to estimate the heritability of MFERT; (2) to estimate genetic correlations between MFERT, traditional fertility traits, and milk production traits; and (3) to assess the potential of MFERT to be used as an indicator trait for fertility in a selection index. The MFERT trait had a heritability of 0.16, which was higher than that (0.05) obtained for traditional fertility traits. Genetic correlations between MFERT and traditional fertility traits were low to moderate. The weakest and strongest correlations (mean ± standard error) were with pregnancy at the end of the mating season (0.13 ± 0.05) and calving to first service (-0.61 ± 0.03), respectively. Based on our estimates, including MFERT in a fertility index will only substantially increase the accuracy of the index when there are many more records available for MFERT than for the traditional fertility traits. This is likely to be the case because the number of milk samples from commercial machines belonging to milk recording companies in Australia that are capable of generating MIR spectra is growing. Hence, the number of records for MFERT is expected to increase substantially in the near future.


An infant intracardiac suction instrument.
An infant intracardiac suction instrument that neither sticks against the heart wall nor obscures the operative field is described.


The Role of the Sinonasal Epithelium in Allergic Rhinitis.
The sinonasal epithelial barrier is comprised of tight and adherens junction proteins. Disruption of epithelial barrier function has been hypothesized to contribute to allergic disease such as allergic rhinitis through increased passage of antigens and exposure of underlying tissue to these stimuli. Several mechanisms of sinonasal epithelial barrier disruption include antigen proteolytic activity, inflammatory cytokine-mediated tight junction breakdown, or exacerbation from environmental stimuli. Mechanisms of sinonasal epithelial barrier stabilization include corticosteroids and nuclear erythroid 2-related factor 2 (Nrf2) cytoprotective pathway activation. Additional studies will aid in determining the contribution of epithelial barrier function in allergic rhinitis pathophysiology and treatment.


Group mean hearing threshold changes in a noise-exposed industrial population using personal hearing protectors.
The audiometric test data of 265 employees in one facility were studied to determine group mean hearing threshold changes. The data covered seven to thirteen years. During this time the population was exposed to workplace noise (OSHA) exposures of 86 to 103 dBA. Personal hearing protection was used. Essentially no change in the mean threshold hearing level was found. Certain other hearing conservation program effectiveness indicators also were used to evaluate this population. The results of these indicators were used to evaluate this population also. The results of these indicators generally follow the trends for effective programs.


Pelvic exenteration for advanced rectal carcinoma.
Twelve patients with advanced rectal cancer and no evidence of extrapelvic metastases underwent pelvic exenteration. The operative mortality rate was 8.3 per cent. Determinate 3 and 5 year survival rates of 54 and 37 per cent were achieved. Criteria for the selection of patients and techniques of supravesical urinary diversion are discussed.


Relationship of the light-induced proton uptake in bovine retinal outer segment fragments to triton-induced membrane disruption and to volume changes.
Light-induced proton uptake in bovine retinal outer segment (ROS) fragments was shown to be closely related to pH, salt concentration, membrane integrity, and perhaps secondarily to the volume of osmotic compartments. The principal findings were as follows: 1. As pH increased, both the discs and the plasmalemma swelled, and proton uptake markedly diminished. 2. As the discs were disrupted by increasing concentrations of Triton, proton uptake at slightly alkaline pH was supplanted by proton release. 3. Increasing the concentration of chloride salts caused increased H+ uptake roughly proportional to osmotic shrinkage of the ROS. Buffering by acetate prevented the measurement of proton uptake in the presence of acetate salts, although osmotic behavior of the ROS was similar to that observed in chloride salts. Although increasing the concentration of sucrose also resulted in osmotic shrinkage of the ROS, it was not accompanied by a systematic increase in the magnitude of proton uptake. 4. Light-induced H+ uptake was accompanied by small but reproducible changes in volume, probably of the discs. The magnitude and direction of these rapid volume changes were subject to influence by pH, solute, and other variables.


Obstacles encountered during transradial angiography from after Radial Artery puncture to the aortic arch.
To elucidate the key points for safe performance of transradial angiography. Transradial angiography can be performed safely if attention is paid to the following points from after radial artery puncture to reaching the aortic arch: resistance during guide wire operation for sheath insertion after puncture; confirmation of the superficial brachial artery; guide wire resistance while guiding the catheter to the aortic arch; and aortic arch anomalies.


Optimization of Chitosan Microspheres Spray Drying via 32 Full Factorial Design.
Generally, the preparation of spray-dried microspheres is strongly affected by the process parameters. Particle size and production yield are mainly influenced by the spraying solution concentration and the pump rate of the spray dryer. The aim of this study was to assess optimum spray drying parameters - polymer concentration and pump rate required for the production of chitosan microspheres with high production yield and targeted for nasal administration particle size. Full 32 factorial design was used to study the investigated parameters. Three different concentrations of the chitosan solution were selected: a low concentration of 1%, average concentration of 1.5% and high concentration of 2%. The rate of the peristaltic pump was also varied at three levels: low rate of 10%, medium rate of 14% and high rate of 18%. Nine models of chitosan microspheres were formulated and characterized in terms of shape, surface morphology, size, particle size distribution and production yield. The particles obtained from 2% chitosan solutions, sprayed at 10% pump rate were of the highest yield (64.33%) and appropriate for nasal administration median diameter (3,434 μm). The two investigated spray-drying parameters interact with each other and their influence on the production yield and the size of the chitosan microspheres should be evaluated together, instead of one at a time. The assessed process parameters allow the production of chitosan microparticles with high yield and desirable characteristics (size, size distribution and shape) for intranasal delivery.


Characterization of early deaths of non- Saccharomyces yeasts in mixed cultures with Saccharomyces cerevisiae.
The survival of Kluyveromyces thermotolerans and Torulaspora delbrueckii in mixed cultures with Saccharomyces cerevisiae was examined at low oxygen availability in a defined grape juice medium. In these fermentations, K. thermotolerans and T. delbrueckii died off earlier than S. cerevisiae, and K. thermotolerans and T. delbrueckii exhibited parabolic death kinetics. Furthermore, the early deaths seemed to be non-apoptotic in nature. In order to understand the mechanism causing the early deaths, various single- and mixed-culture fermentations were carried out. The early deaths could not be explained by nutrient depletion or the presence of toxic compounds. Rather, they seemed to be mediated by a cell-to-cell contact mechanism at high cell densities of S. cerevisiae, and to a lesser ability of K. thermotolerans and T. delbrueckii to compete for space, as compared to S. cerevisiae. These results contribute to an increased understanding of why K. thermotolerans and T. delbrueckii die off before S. cerevisiae in wine fermentations.


Primary and challenge infection of mice with equine herpesvirus 1, strain HSV25A.
Clinical signs, haematology, lymphocyte subset analysis, viral clearance, lung histopathology and humoral and cell-mediated (CMI) immune responses were monitored throughout the acute and convalescent phases of infection in groups of BALB/c mice infected intranasally with equine herpesvirus 1 (EHV-1), strain HSV25A. Primary infection caused a leucocytosis due to a neutrophilia during days 1 and 2 post-infection (pi) and a B lymphocytosis at day 1 pi. Serum ELISA antibodies were detected by 7 days pi and neutralising antibodies by 2 weeks pi. Mice infected with EHV-1 were not protected against disease when challenged with EHV-1 12 weeks later. However, viral clearance from lungs was significantly faster and the antibody response was markedly enhanced within the first few days of challenge infection. A CMI response was detected by 5 days after primary infection, but the level of responsiveness was not increased by challenge infection, although the lungs of challenged mice had markedly increased numbers of mononuclear cells around blood vessels and bronchioles. Specific antibodies to glycoprotein (g) B were detected by 2 weeks pi, 4 weeks earlier than the detection of antibodies to gC and 10 weeks before those to gD. The primary response was relatively short-lived with neither ELISA antibody nor lymphocyte proliferation was evident by 6 months pi.


Management of Traumatic Atlantooccipital Dissociation at a Level 1 Trauma Center: A Retrospective Case Series.
Atlantooccipital dislocation (AOD) is a highly unstable and often neurologically devastating injury to the craniocervical junction that typically results from high-energy trauma. Management of these devastating injuries is complex, with prognostication difficult due to high rates of concomitant intracranial and systemic injuries. This report highlights advances in management of AOD and appropriate implementation of operative adjuncts including neuronavigation and the use of intraoperative neuromonitoring. All patients with AOD presenting to a high-volume, level 1 trauma center between January 2015 and August 2021 were retrospectively identified through a prospectively maintained database of patients presenting with traumatic spine injuries. Medical records, including imaging reports, clinical documentation, and intraoperative neurophysiological reports were reviewed. A total of 11 patients were identified with patterns of injury consistent with AOD. Fifty-five percent of patients survived until discharge. 73% of patients underwent surgery for stabilization. All 4 patients with preoperative neurologic deficits who underwent surgery had monitorable transcranial motor evoked potentials and somatosensory evoked potentials. Two experienced significant motor recovery postoperatively, and 2 did not survive to discharge. Blunt cerebrovascular injuries were identified in 73% of patients. AOD is encountered with increasing frequency. The identification and management of this specific injury is complicated by the volume and severity of associated injuries, especially concomitant traumatic brain injury. Timely recognition is critical and the use of surgical adjuncts including intraoperative neurophysiologic monitoring and surgical navigation can increase the safety and success of these procedures while also providing prognostic information on potential for motor recovery.


Glycogen synthase kinase-3 interacts with and phosphorylates estrogen receptor alpha and is involved in the regulation of receptor activity.
Like other steroid hormone receptors, estrogen receptor-alpha (ERalpha) is a substrate for protein kinases, and phosphorylation has profound effects on the function and activity of this receptor. A number of different kinases have been implicated in ERalpha regulation. In this report we show by mutational analysis and in vitro kinase assays that ERalpha is a substrate for glycogen synthase kinase-3 (GSK-3) in vitro and is phosphorylated on two sites, the Ser-102, -104, and -106 motif and Ser-118, both located in the N-terminal transcription activation function (AF-1) domain. GSK-3 forms a complex with ERalpha in vivo as demonstrated by co-immunoprecipitation from cell lysates. The GSK-3 inhibitor lithium chloride was used to determine the role of GSK-3 in phosphorylation of Ser-102, -104, and -106 and Ser-118 in vivo and to explore the role of these serines in the regulation of ERalpha function. Treatment of cells with lithium chloride resulted in dephosphorylation of Ser-104 and -106 and Ser-118, which suggests these serine residues as targets for GSK-3 in vivo. Our results further suggest that ERalpha phosphorylation by GSK-3 stabilizes ERalpha under resting conditions and modulates ERalpha transcriptional activity upon ligand binding. Inhibition and constitutive activation of GSK-3, both, resulted in inhibition of ERalpha transcriptional activity, indicating a function of active as well as inactive GSK-3 in ERalpha regulation. These findings uncover a novel mechanism for the regulation of ERalpha-mediated estrogen signaling controlled by a dual action of GSK-3.


Can arguments address concerns?
People have concerns, and ethicists often respond to them with philosophical arguments. But can conceptual constructions properly address fears and anxieties? It is argued in this paper that while it is possible to voice, clarify, create and-to a certain extent-tackle concerns by arguments, more concrete practices, choices, and actions are normally needed to produce proper responses to people's worries. While logical inconsistencies and empirical errors can legitimately be exposed by arguments, the situation is considerably less clear when it comes to moral, cultural, and emotional norms, values, and expectations.


Significance of hemorrhagic endovasculitis in placentae from stillbirths.
Hemorrhagic endovasculitis (HEV) is an alteration of fetal chorionic blood vessels within the placenta. This disruption of the integrity of the parietal blood vessels has been reported in association with poor pregnancy outcomes and perinatal complications. In stillbirths, its presence has been implicated as part of the mechanism of the cessation of fetal circulation in postmortem retention of the placenta. The purpose of this study was to give an interpretation of this type of blood vessel damage: whether it is related to the arrest of blood circulation after stillbirth, or whether it constitutes an alteration of the integrity of placental chorionic circulation contributing to the death of the fetus. From 1232 reviewed placentae, we analyzed 46 cases with HEV, 70% from livebirths and 30% from stillbirths. The most frequent lesions encountered in placentae were categorized microscopically. The distribution of these lesions in placentae from livebirth and stillbirth pregnancies was then described. In association with HEV, villitis of unknown etiology and chorionic vessel thrombi occurred more frequently in placentae from livebirths. Fragmented erythrocytes and erythroblasts were more frequent in placentae from stillbirths. Our study demonstrates that HEV is more consistently associated with other placental lesions in liveborn than in stillborn infants.


A subjective analysis of participants' views in a study of experimental modelling of imposed airway obstruction in infants and children.
Deliberate airway obstruction is a well-recognised form of child abuse and one of the most common causes of death in cases of child homicide. Although classical signs such as petechial haemorrhages can be helpful in raising the suspicion of mechanical asphyxia, they are not always present. Therefore, distinguishing between bruising caused by accidents, medical conditions or non-accidental injury remains challenging. We aimed to characterise bruising patterns which are potentially more consistent with deliberate airway obstruction by means of an experimental modelling study. The first results of our project were previously published and demonstrated that bruising anywhere on the head may be consistent with deliberate upper airway obstruction. In this paper, we present the findings of a questionnaire carried out during the modelling to assess participants' perception of force and consider how variables such as age, use of adjuncts and force distribution may affect bruising patterns in deliberate airway obstruction. Statistical analysis of our results showed that participants felt they were using less force in scenarios involving the infant rather than the child resuscitation dummy as well as when using adjuncts, meaning marks are likely to be more subtle. Therefore, in such cases it is important to examine for other signs of asphyxiation (such as petechiae) and consider the possibility that adjuncts might have been used which could make picking up localised injuries more difficult. Our results also showed that participants often felt force was not distributed evenly across the digits, reporting the greatest force through either the thumb in isolation or the thumb plus another digit in up to nearly 50% of cases. This suggests that just one or two bruises may be consistent with deliberate airway obstruction.


Autoantibodies against nuclear envelope-associated proteins in primary biliary cirrhosis.
An antinuclear immunofluorescence pattern displaying a thin ring confined to the nuclear envelope was assessed in sera from 38 patients with primary biliary cirrhosis and in sera from a control group of 277 patients with other antinuclear antibody-positive diseases. This rim-like antinuclear reactivity was present in sera from 20 primary biliary cirrhosis patients (52.6%) but in only two patients from the control group (0.7%) (p less than 0.001). Furthermore, this autoantibody was present in three of four primary biliary cirrhosis patients without antimitochondrial antibodies. Presence of this rim-like pattern in primary biliary cirrhosis did not correlate with the presence of associated autoimmune diseases nor with other clinical, biochemical, nor histological features of the disease. The antigenic specificity of sera displaying this antinuclear immunofluorescence pattern was characterized by Western blot analysis using an antigenic extract containing nuclear envelope proteins purified from rat liver. Sixteen of the 20 positive sera showed a common pattern of reactivity with a set of nuclear envelope-associated proteins approximately 200 kD in size. In conclusion, sera from primary biliary cirrhosis patients showing a rim-like fluorescent nuclear pattern have antinuclear autoantibodies that react specifically with components of the nuclear envelope. The high specificity of these new autoantibodies in primary biliary cirrhosis indicate that they might be a serological marker of the disease, particularly useful in patients without antimitochondrial antibodies.


The sodium gradient induces conformational changes in the renal phosphate carrier.
Phosphate transport across brush border membranes from kidney cortex is very sensitive to inhibition by phenylglyoxal, an arginine modifier. Sodium-dependent phosphate influx into brush border membrane vesicles was inhibited by 60%. In contrast, phenylglyoxal had no effect on passive influx or on sodium-dependent efflux of phosphate. Preincubation of the vesicles with sodium prior to the addition of phenylglyoxal demonstrated a strong protective effect of intravesicular sodium (73% protection). Phosphate also protected the transporter from inhibition, but from the extravesicular side only (63%). Substitution of phosphate by sulfate offered no protection at all, indicating the specificity of protection. Addition of both substrates (sodium and phosphate) offered an additional protection from the extravesicular side compared to that offered by phosphate alone (92 versus 55%). There was no additional protection when both substrates were added to the intravesicular side. Phosphate influx measured in the presence of sodium but in the absence of a sodium gradient was totally unaffected by phenylglyoxal modification. There was no inhibition on phosphate influx measured in equilibrium exchange conditions. We propose a model for the phosphate carrier in which the sodium gradient induces a conformational change and an arginine residue is essential for the coupled flux of sodium and phosphate.


To screen, or not to screen,--that is the question.
Recent studies of school dental screening and general dental practice in the North West of England may have provided an evidence base for the review of dental school screening and its improvement by increasing: the relevance of referral criteria, the rate of attendance resulting from screening, and treatment rates following referral--rather than for abandoning the process altogether.


Strand-specific, real-time RT-PCR assays for quantification of genomic and positive-sense RNAs of the fish rhabdovirus, Infectious hematopoietic necrosis virus.
The fish rhabdovirus, Infectious hematopoietic necrosis virus (IHNV), is an important pathogen of salmonids. Cell culture assays have traditionally been used to quantify levels of IHNV in samples; however, real-time or quantitative RT-PCR assays have been proposed as a rapid alternative. For viruses having a single-stranded, negative-sense RNA genome, standard qRT-PCR assays do not distinguish between the negative-sense genome and positive-sense RNA species including mRNA and anti-genome. Thus, these methods do not determine viral genome copy number. This study reports development of strand-specific, qRT-PCR assays that use tagged primers for enhancing strand specificity during cDNA synthesis and quantitative PCR. Protocols were developed for positive-strand specific (pss-qRT-PCR) and negative-strand specific (nss-qRT-PCR) assays for IHNV glycoprotein (G) gene sequences. Validation with synthetic RNA transcripts demonstrated the assays could discriminate the correct strand with greater than 1000-fold fidelity. The number of genome copies in livers of IHNV-infected fish determined by nss-qRT-PCR was, on average, 8000-fold greater than the number of infectious units as determined by plaque assay. We also compared the number of genome copies with the quantity of positive-sense RNA and determined that the ratio of positive-sense molecules to negative-sense genome copies was, on average, 2.7:1. Potential future applications of these IHNV strand-specific qRT-PCR assays are discussed.


The association of constipation with childhood urinary tract infections.
Previous studies have suggested a correlation between constipation and urinary tract infections (UTIs) in children. However, historical information about constipation may be unreliable and the relationship between a history of constipation and radiographic findings of fecal load is unclear. A total of 133 children undergoing an abdominal X-ray were evaluated. Parents were asked to complete a questionnaire on bowel habits. Three observers using a documented objective scoring system evaluated plain films of the abdomen. The symptom and radiographic scores were compared with the history and each other. Out of the 133 children, 100 had documented previous UTIs and 33 did not. Children with prior UTIs had significantly more symptoms of constipation than those without prior UTIs (p<0.02). Children with a history of UTIs tended to have more fecal loading on radiographic studies than those without, although this difference was not statistically significant (p<0.11). When only children of >3 years old are evaluated, the trends persist, but neither were statistically significant (p<0.11 and 0.56, respectively). There was a poor correlation between the symptoms of constipation and fecal load on abdominal X-rays (correlation coefficient of 0.08). Our findings support the concept that children with UTIs have a higher rate of constipation, especially by history. However, diagnosing constipation in individual patients is difficult. Not only is there a poor correlation between history and radiographic findings of constipation in any individual patient, but at this time there is no gold standard.


Current role of cryotherapy in retinopathy of prematurity: a report by the American Academy of Ophthalmology.
To evaluate the role of cryotherapy in the current treatment of retinopathy of prematurity (ROP). Literature searches of PubMed and the Cochrane Library were conducted on December 2, 2009, for articles published after 1984. The searches included all languages and retrieved 187 relevant citations. Thirteen articles were deemed relevant to the assessment question and were rated according to the strength of evidence. Four articles reported results from 2 large multicenter randomized clinical trials, and the remaining 9 articles reported results of 3 small randomized trials that directly compared cryotherapy and laser. Neither of the multicenter randomized clinical trials was a direct comparison of cryotherapy with laser. These studies were used to evaluate the comparative trials based on treatment criteria, study populations, and clinical results. Higher percentages of poor structural and functional outcomes generally were seen in eyes treated with cryotherapy compared with eyes undergoing laser treatment. Higher rates of systemic complications and myopia also were identified after treatment with cryotherapy. Despite a relative paucity of level I evidence directly comparing cryotherapy and laser treatment for threshold ROP, the literature suggests that neonatal facilities should gain access to laser technology and laser-trained ophthalmic staff to achieve better outcomes for treatment of the disease.


Collective variables for the study of long-time kinetics from molecular trajectories: theory and methods.
Collective variables are an important concept to study high-dimensional dynamical systems, such as molecular dynamics of macromolecules, liquids, or polymers, in particular to define relevant metastable states and state-transition or phase-transition. Over the past decade, a rigorous mathematical theory has been formulated to define optimal collective variables to characterize slow dynamical processes. Here we review recent developments, including a variational principle to find optimal approximations to slow collective variables from simulation data, and algorithms such as the time-lagged independent component analysis. Using these concepts, a distance metric can be defined that quantifies how slowly molecular conformations interconvert. Extensions and open questions are discussed.


Campylobacter hyointestinalis subsp. hyointestinalis, a common Campylobacter species in reindeer.
To study the prevalence of Campylobacter spp. in the faecal material of reindeer, and to identify the isolates by means of a polyphasic approach. In addition, to study the genetic diversity of Camp. hyointestinalis subsp. hyointestinalis reindeer isolates by pulsed-field gel electrophoresis (PFGE). The material, collected during the slaughter period in autumn 1998, comprised 399 faecal contents from the reindeer (Rangifer tarandus), a semi-domesticated, meat-producing ruminant of northern Finland. These samples came from 16 herds in the areas of eight reindeer slaughterhouses. Samples were cultured by methods suitable for isolation of fastidious Campylobacter species. Of all samples, 6% (24/399) were Campylobacter-positive. Phenotypic characteristics, SDS-PAGE protein patterns, dot blot DNA-DNA hybridization, 23S rDNA restriction fragment polymorphism analysis and PFGE identified the isolates as Camp. hyointestinalis subsp. hyointestinalis. Campylobacter hyointestinalis subsp. hyointestinalis was the only Campylobacter species isolated from reindeer in this study. The isolates showed high genomic diversity in PFGE with the restriction enzymes SmaI and KpnI. PFGE analysis is a useful subtyping method for epidemiological studies. Contaminated reindeer meat can be a source for human infections.


Nocturnal prolactin and cortisol secretion and recovery from melancholia.
Spontaneous prolactin and cortisol patterns were determined at 20 min intervals over 3 hr during the night in eight patients with melancholia, both during illness and after treatment with amitriptyline. Mean plasma prolactin levels were greater after recovery in the seven patients who responded to treatment. Mean cortisol secretion decreased upon recovery from melancholia, and such changes in two patients paralleled normalization of dexamethasone suppression test responses. The influence of assumptions of lack of interaction on the statistical significance of the analysis of variance with repeated measures for prolactin and cortisol values was evaluated.


Estimation of Cost for Endoscopic Screening for Esophageal Cancer in a High-Risk Population in Rural China: Results from a Population-Level Randomized Controlled Trial.
Population-level endoscopic screening for esophageal cancer has been conducted in China for years. In this study, we aim to provide an updated and precise cost estimation for esophageal cancer screening based on a randomized controlled trial in a high-risk area in China. We estimated the cost of esophageal cancer screening with chromoendoscopy using a micro-costing approach based on primary data of the ESECC (Endoscopic Screening for Esophageal Cancer in China) randomized controlled trial (NCT01688908) from a health sector perspective. Unit costs and quantities of resources were collected to obtain annual screening costs. The screening project was then theoretically expanded to a 10-year period to explore long-term trends of costs. Costs were adjusted to US dollars for the year 2018. In the ESECC trial, screening cost per endoscopy with a valid pathologic diagnosis was $196, accounting for 3.82% of the gross domestic product per capita in Hua County, and the costs for detecting one esophageal cancer and one early-stage esophageal cancer were $26,347 and $37,687, respectively. In conventional screening in which protocol-driven costs were excluded, costs as above were $134, $18,074, and $25,853. The cost for detecting one gastric cardia cancer or stomach cancer was nine times higher than detecting one esophageal cancer owing to low prevalences of the two cancers. In a simulated 10-year screening project, annual cost decreased notably over time. Despite the relatively low absolute cost, population-level endoscopic screening will still be a heavy burden on local government considering the socioeconomic conditions. Long-lasting programs would be less costly and population-level screening would make little sense in non-high-risk regions.


Proximal junctional kyphosis and failure-diagnosis, prevention, and treatment.
Technical advancements have enabled the spinal deformity surgeon to correct severe spinal mal-alignment. However, proximal adjacent segment pathology (ASP) remains a significant issue. Examples include proximal junctional kyphosis (PJK) and proximal junctional failure (PJF). Agreement on the definition, classification, and pathophysiology of PJK and PJF remains incomplete, and an understanding of the risk factors, means of prevention, and treatment of this problem remains to be elucidated. In general, PJK is a relatively asymptomatic radiographic diagnosis managed with patient reassurance and monitoring. On the other hand, PJF is characterized by mechanical instability, pain, and more severe kyphosis, with potential for neurologic compromise. Patients who develop PJF more often require revision surgery than those with PJK. This chapter will review the current understanding of PJK and PJF.


Prevention of systemic lupus erythematosus-like disease in (NZBxNZW)F1 mice by treating with CDR1- and CDR3-based peptides of a pathogenic autoantibody.
Two peptides based on the complementarity-determining regions (CDR) of a pathogenic murine anti-DNA antibody were employed in an attempt to prevent the spontaneous systemic lupus erythematosus (SLE)-like disease of (NZBxNZW)F1 mice. Female mice, at the age of 2 months, were injected with either the CDR1- or the CDR3-based peptides (pCDR1, pCDR3) subcutaneously or intravenously in aqueous solution for a total of 8-10 treatments. A reduction was observed in the total and pathogenic IgG2a and IgG3 anti-DNA antibody titers in the CDR-treated groups. Treatment reduced the number of mice that developed proteinuria and immune complex deposits in their kidneys. The severity of renal pathology was significantly reduced in the pCDR3 (P<0.02) and pCDR1 (P< or = 0.05) treated mice. Thus, both CDR-based peptides administered in aqueous solution were capable of preventing the SLE-like disease in (NZBxNZW)F1 mice, although the beneficial effects of pCDR3 appeared to be more pronounced than those of pCDR1 in the treated mice.


The treatment of elopement behavior in a retardate using a graduated levels program.
The present study investigated whether a levels program would effectively reduce the amount of elopement (running-away) behavior demonstrated by an adult with profound mental retardation. The subject resided in an Alternative Intermediate Services (AIS) group home. During baseline data collection, he demonstrated an average of 131 elopements per month. After implementation of the levels program, elopements decreased to 0 within 7 months. At the end of the study, the subject's elopements continued to be maintained at a very low rate.


N-fragment of edema factor as a candidate antigen for immunization against anthrax.
The nontoxic N-terminal fragment of Bacillus anthracis edema factor (EF) was evaluated as a candidate antigen in an anthrax vaccine using a replication-incompetent adenoviral vector. An E1/E3 deleted adenovirus (Ad/EFn) encoding the N-terminal region 1-254 amino acids of the edema factor (EFn) was constructed using the native DNA sequence of EFn. Intramuscular immunization three times with 10(8) plaque forming units (pfu)/dose of Ad/EFn in A/J mice resulted in 37% and 57% protection against a subcutaneous challenge with B. anthracis Sterne strain spores at a dosage of 200 x LD50 and 100 x LD50, respectively. EF-specific serum IgG responses (including total IgG, IgG1, and IgG2a isotype titers) were robust in the Ad/EFn immunized animals. Interestingly, anti-EF antibodies cross-reacted with anthrax lethal factor (LF), and had a neutralizing capability against both anthrax lethal toxin (Letx) and edema toxin (Edtx), as demonstrated by in vitro toxin neutralization assays using J774A.1 mouse macrophage and Chinese hamster ovary cell (CHO), respectively. Our data suggest that EF plays a role in eliciting protective immunity against anthrax, and that it should be included in a new generation multi-component subunit vaccine.


A ten-year prospective study on gallbladder stone recurrence after successful extracorporeal shock-wave lithotripsy.
The risk of recurrence has limited the acceptability of conservative therapies of gallbladder stones. The aim of the present study was to determine the long-term rate of stone recurrence and its risk factors after successful extracorporeal shock-wave lithotripsy (ESWL). The study comprised a prospective ultrasound follow-up at yearly intervals or whenever biliary pain was reported. A total of 192 consecutive patients (primary single stones, n=159; primary 2 or 3 stones, n=33) were followed for up to 11.2 years after becoming stone-free and after termination of adjuvant treatment with ursodeoxycholic acid (UDCA). Eighty-four patients developed recurrent stones after a median of 2.6 years (maximum?=?8.8 years). The 108 patients without recurrence were followed for a median of 6.7 years (maximum=11.2 years). By actuarial analysis, the cumulative recurrence rates for these 192 stone-free patients were 27%+/-3%, 41%+/-4% and 54%+/-4% (observed +/-SE) at 3, 5 and 10 years, respectively. Cox's regression analysis was used to identify the presence of slight calcification in the primary stone(s) as a protective feature against recurrence (p=0.03). 1) The risk of recurrence continues to increase over time, and although it rises less steeply after 5 years, it does not reach a plateau until at least 10 years. 2) Having had slightly calcified stone(s) seems to be associated with a reduced risk of recurrence and might signal a "burnt out" lithogenic process. 3) The long-term results are unsatisfactory and ESWL of gallbladder stones should be offered only in special cases.


Heptamer-type small guide RNA that can shift macrophages toward the M1 state.
Multiple myeloma is a refractory cancer of plasma cells. Although treatment strategies for multiple myeloma are getting improved year by year, in most cases patients relapse due to the emergence of drug-resistant mutations in the myeloma cells. The interplay between myeloma cells and tumor-associated macrophages (TAM) is important for the pathology. We thought that some heptamer-type sgRNAs for TRUE gene silencing would be able to transform TAM toward the M1 state and might become therapeutic drugs for myeloma. Here, we searched for heptamer-type sgRNAs that can shift macrophages toward the M1 state. We screened a heptamer-type sgRNA library for the ability to up-regulate IL-12b gene expression in human macrophage-like cell lines, and found three such sgRNAs. One of the sgRNAs, H12960, which also showed such ability in human fresh macrophages and mouse macrophage-like cell lines, efficiently suppressed human myeloma cell growth in SCID/NOD mice.


Primary tonsillar tuberculosis masquerading as carcinoma of the tonsil.
Primary tuberculosis (TB) of tonsil is a rare form of extra-pulmonary tuberculosis. Most tonsillar TB cases present with coexistent pulmonary tuberculosis. It can simulate tonsillar malignancy and poses a diagnostic challenge. Histopathological examination is often needed for confirmation. Herein, we report a case of primary tonsillar tuberculosis in a 55-year-old gentleman mimicking carcinoma of the tonsil.


[« The desire to know where I came from pushed me to start the research »].
This interview retraces the journey of a man who, wishing to know where he came from, decided to go in search of his origins. From wanderings to hesitations, from bitterness to determination, Arthur Kermalvezen Fournis describes the different steps that led him to the truth. A painful but salutary fight.


Prevalence of Oppositional Defiant Disorder in a sample of Spanish children between six and sixteen years: teacher's report.
Our main objective is to study the prevalence of Oppositional Defiant Disorder (ODD) in school children aged 6-16 years of an autonomous region of Spain (Castile and Leon), according to reports from the teachers and to analyze the impact of the disorder on academic performance and school behavior. Population study with stratified multistage, proportional and cluster design sample. Sample analyzed: 1,049. Cases were defined according to DSM-IV-TR criteria. An overall prevalence rate of 4.2% was found, this being significantly higher in males (5.7%) compared to females (2.6%) and in rural (6.8%) than in urban areas (3%). No significant differences by grade or type of school were found. ODD prevalence without considering functional impairment would increase to 5.1%. ODD cases have significantly worse academic outcomes (overall academic performance, reading, math and writing) and entail worse classroom behavior (relationship with peers, respect for rules, organizational skills, academic tasks and disruption of the class). Based on the prevalence, early onset, persistence of symptoms and social and academic dysfunction of ODD, early diagnosis and preventive intervention are necessary.


Imbalance of angiogenic factors and avascular edematous cystic villi in a trisomy 13 pregnancy: a case report.
The incidence of pre-eclampsia is significantly higher in trisomy 13 pregnancies than in normal pregnancies. Soluble fms-like tyrosine kinase-1 (sFlt-1), located on chromosome 13, is an anti-angiogenic molecule derived from the placenta and contributes to the pathogenesis of pre-eclampsia. Elevated sFlt-1 and reduced placental growth factor (PlGF) are associated with trisomy 13 pregnancies and may play a pathogenic role in the subsequent development of pre-eclampsia. Here we present a case of a trisomy 13 pregnancy without any signs of pre-eclampsia that showed alterations in circulating angiogenic factors and abnormal placental appearance. The placenta developed edematous changes and contained multiple small cysts. Histology of the placenta confirmed avascular edematous cystic villi and did not show the typical appearance of a partial mole or mesenchymal dysplasia. The sFlt-1/PlGF ratio in maternal serum (134) was much higher than that in gestational age-matched women who were normotensive (2.9-7.2; mean, 5.0). Immunostaining for Flt-1 and endoglin was more intense in our case compared with gestational age-matched controls, and at a similar level to a case of pre-eclampsia. Placental findings that showed avascular edematous cystic villi in our case may be associated with angiogenic imbalance involved in the pathogenesis of pre-eclampsia in trisomy 13 pregnancies.


Protocol for optimizing production and quality control of infective EcoHIV virions.
EcoHIV is a model of HIV infection that recapitulates aspects of HIV-1 pathology in mice. However, there are limited published protocols to guide EcoHIV virion production. Here, we present a protocol for producing infective EcoHIV virions and essential quality controls. We describe steps for viral purification, titering, and multiple techniques to analyze infection efficacy. This protocol produces high infectivity in C57BL/6 mice which will aid investigators in generating preclinical data.


Increased ovarian tumor cell surface reacting antibodies in patients with ovarian adenocarcinoma after viral oncolysate treatment.
Antibodies reacting with the cell surface of ovarian tumor cell lines were detected in the sera of untreated patients with ovarian cancer using cell surface immunofluorescence and FACS analysis. Vaccination of these patients with viral oncolysates increased the antibody production against ovarian cell surface antigens. These antibodies cross-reacted with a melanoma cell line (WM-75), and a lymphoblastoid cell line (Daudi). Immunoprecipitation and SDS-PAGE analysis of 125I-iodine labelled cell surface antigens of ovarian cells revealed a response to private and common antigens expressed on the cell surface of the tumor cells.


Histopathological classification of lesions observed in natural cases of paratuberculosis in free-ranging fallow deer (Dama dama).
Ninety-five adult fallow deer, legally hunted in the Regional Hunting Reserve of El Sueve (Northern Spain), were subjected to a post-mortem examination for paratuberculosis, samples being taken from the proximal and distal jejunum, proximal and distal ileum, ileocaecal valve and associated lymph nodes. The lesions were divided into four categories. Focal lesions (n=19 cases) consisted of small granulomas, mainly in the jejunal and ileal lymph nodes. Multifocal lesions (n=4) consisted of well-demarcated granulomas in the intestinal lymphoid tissue and also in the intestinal lamina propria. Diffuse multibacillary lesions (n=2) were characterized by a severe granulomatous enteritis and lymphadenitis. Macrophages and numerous Langhans giant cells containing many mycobacteria were present, resulting in macroscopical changes in the normal gut morphology. These changes were found from the proximal jejunum to the ileocaecal valve, but lesions were always particularly severe in the distal jejunum. In diffuse intermediate (multibacillary-lymphocytic) lesions (n=3) the infiltrate consisted of lymphocytes, macrophages and Langhans giant cells, with small numbers of mycobacteria. Mycobacterium avium subspecies paratuberculosis was identified by a polymerase chain reaction technique. The widespread occurrence of paratuberculosis in fallow deer in this Reserve represents a potential source of infection for other susceptible species.