Immobilization of oligonucleotides with homo-oligomer tails onto amine-functionalized solid substrates and the effects on hybridization.
Microarrays have become important tools for the detection and analysis of nucleic acid sequences. Photochemical (254 nm UV) DNA immobilization onto amine-functionalized substrates is often used in microarray fabrication and Southern blots, although details of this process and their effects on DNA functionality are not well understood. By using Cy5-labeled model oligonucleotides for UV immobilization and Cy3-labeled complementary sequences for hybridization, we measured independently the number of immobilized and hybridized oligonucleotides on the microarray surface. By using a two-color fluorescence LED setup and a novel method to compile the data, a full analysis has been made of the effects of oligonucleotide composition (length and sequence) on both immobilization and hybridization. Short homo-oligomer sequences (tails) of uracils, thymines, and, to a limited extent, guanines attached to a hybridization sequence improve immobilization. We propose a possible mechanism explaining the grafting of these nucleotides to amine-functionalized substrates, and we found evidence that the DNA backbone is possibly involved in the immobilization process. Hybridization, on the other hand, greatly improves as a function of tail length regardless of tail composition. On the basis of statistical arguments, the probes increasingly bind via their tail, with the hybridization sequence becoming more accessible to its complement. We conclude that all tails, sequence independent, improve hybridization signals, which is caused by either improved immobilization (especially thymine and uracil) or improved hybridization (most pronounced with guanine tails).


Synaptamide modulates glial and neurotransmitter activity in the spinal cord during neuropathic pain.
N-docosahexaenoylethanolamine, or synaptamide, is an endogenous metabolite of docosahexaenoic acid that is known for synaptogenic and neurogenic effects. In our previous studies we have shown that synaptamide attenuates neuropathic pain, facilitates remyelination, and reduces neuroinflammation after the chronic constriction injury (CCI) of the sciatic nerve in rats. In the current study, we show that daily synaptamide administration (4 mg/kg/day) within 14 days post-surgery: (1) decreases micro- and astroglia activity in the dorsal and ventral horns of the lumbar spinal cord; (2) modulates pro-inflammatory (IL1β, IL6) and anti-inflammatory (IL4, IL10) cytokine level in the serum and spinal cord; (3) leads to a rise in synaptamide and anandamide concentration in the spinal cord; (4) enhances IL10, CD206 and N-acylethanolamine-hydrolyzing acid amidase synthesis in macrophage cell culture following LPS-induced inflammation. Thus, the ability of synaptamide to modulate glial and cytokine activity indicates its potential for implementation in the treatment peripheral nerve injury.


Efficient Synthesis of Cannabigerol, Grifolin, and Piperogalin via Alumina-Promoted Allylation.
The synthesis of three phenolic natural products has been accomplished with unprecedented efficiency using a new alumina-promoted regioselective aromatic allylation reaction. Cannabigerol and grifolin were prepared in one step from the inexpensive 5-alkyl-resorcinols olivetol and orcinol. Piperogalin was synthesized, for the first time, via two sequential allylations of orcinol with geraniol and prenol.


Prognostic and Biologic Relevance of Clinically Applicable Long Noncoding RNA Profiling in Older Patients with Cytogenetically Normal Acute Myeloid Leukemia.
We have previously shown that expression levels of 48 long noncoding RNAs (lncRNA) can generate a prognostic lncRNA score that independently associates with outcome of older patients with cytogenetically normal acute myeloid leukemia (CN-AML). However, the techniques used to identify and measure prognostic lncRNAs (i.e., RNA sequencing and microarrays) are not tailored for clinical testing. Herein, we report on an assay (based on the nCounter platform) that is designed to produce targeted measurements of prognostic lncRNAs in a clinically applicable manner. We analyzed a new cohort of 76 older patients with CN-AML and found that the nCounter assay yielded reproducible measurements and that the lncRNA score retained its prognostic value; patients with high lncRNA scores had lower complete remission (CR) rates (P = 0.009; 58% vs. 87%), shorter disease-free (P = 0.05; 3-year rates: 0% vs. 21%), overall (OS; P = 0.02, 3-year rates: 10% vs. 29%), and event-free survival (EFS; P = 0.002, 3-year rates: 0% vs. 18%) than patients with low lncRNA scores. In multivariable analyses, the lncRNA score independently associated with CR rates (P = 0.02), OS (P = 0.02), and EFS (P = 0.02). To gain biological insights, we examined our initial cohort of 71 older patients with CN-AML, previously analyzed with RNA sequencing. Genes involved in immune response and B-cell receptor signaling were enriched in patients with high lncRNA scores. We conclude that clinically applicable lncRNA profiling is feasible and potentially useful for risk stratification of older patients with CN-AML. Furthermore, we identify potentially targetable molecular pathways that are active in the high-risk patients with high lncRNA scores.


Cervical cancer trends in the United States: a 35-year population-based analysis.
To analyze trends in invasive cervical cancer incidence by age, histology, and race over a 35-year period (1973-2007) in order to gain insight into changes in the presentation of cervical cancer. Data from the nine Surveillance, Epidemiology, and End Results (SEER) registries that continuously collected information on invasive cervical cancer were analyzed for trends. Standardized to the 2000 U.S population, annual age-adjusted incidence rates were estimated by race and histologic subtype. Histologic subtype was classified into squamous, adenocarcinoma, and adenosquamous. Overall incidence rates for invasive cervical cancer decreased by 54% over the 35 years, from 13.07/100,000 (1973-1975) to 6.01/100,000 (2006-2007), and the incidence rates declined by 51% and 70.2%, respectively, among whites and blacks. The incidence rates for squamous carcinoma decreased by 61.1% from 10.2/100,000 (1973-1975) to 3.97/100,000 (2006-2007). Incidence rates for adenosquamous cell carcinomas decreased by 16% from 0.27/100,000 (1973-1975) to 0.23/100,000 (2006-2007), and incidence rates for adenocarcinomas increased by 32.2% from 1.09/100,000 (1973-1975) to 1.44/100,000 (2006-2007). This increase in adenocarcinomas was due to an increase in incidence in white women; a decrease in incidence was observed for black women. Although marked reductions in the overall and race-specific incidence rates of invasive cervical cancer have been achieved, they mask important variation by histologic subtype. These findings suggest that alternatives to Pap smear-based screening, such as human papillomavirus (HPV) testing and HPV vaccination, need to be prioritized if adenocarcinomas of the cervix are to be controlled.


Retroperitoneal fibrosis; a single-centre case experience with literature review.
We present 13 patients with retroperitoneal fibrosis, focusing on clinical features, radiological characteristics, treatments and their outcomes. Retrospective review of the medical records was performed of all retroperitoneal fibrosis patients diagnosed and treated in our department between 2012 and 2017. Twelve patients were male, with a median age of 64 years. Eleven patients presented with abdominal pain or back pain or both. Aetiologies varied from idiopathic to malignancy and vasculitis. Twelve patients had PET scans. These showed 18F-fluorodeoxyglucose-avid retroperitoneal soft tissue around the abdominal aorta in the vast majority, with five scans also demonstrating localized or generalized uptake by the aorta. In all cases except one, glucocorticoids were applied as the first-line therapy. Further immunosuppressive therapy was required in 10 cases. Our patients were male and older in age compared with the existing literature. PET scans were very helpful in diagnosis of retroperitoneal fibrosis. Rituximab was found to be an effective treatment in six of our patients.


Using stand/sit workstations in classrooms: lessons learned from a pilot study in Texas.
Childhood obesity has grown into a national epidemic since the 1980s. Many school-based intervention efforts that target childhood obesity involve curriculum and programming that demands instructional time, which disincentivizes school participation. Stand-biased classrooms are an environmental intervention that promotes standing rather than sitting by utilizing standing height desks that allow students to stand during normal classroom activities. The quasi-experimental pilot study was conducted in 5 first-grade classrooms in a Texas elementary school, with 2 control classrooms, 2 treatment classrooms, and 1 classroom that was a control in the fall and treatment in the spring (to allow for within-group comparisons). This intervention has been shown effective in significantly increasing caloric expenditure. In addition, the present study reveals potential behavioral effects from standing. This article presents lessons learned from the pilot study that may prove useful for others implementing similar interventions and calls for additional research on the academic benefits of standing for students.


Antibody responses to Bordetella pertussis Fim2 or Fim3 following immunization with a whole-cell, two-component, or five-component acellular pertussis vaccine and following pertussis disease in children in Sweden in 1997 and 2007.
Bordetella pertussis fimbriae (Fim2 and Fim3) are components of a five-component acellular pertussis vaccine (diphtheria-tetanus-acellular pertussis vaccine [DTaP5]), and antibody responses to fimbriae have been associated with protection. We analyzed the IgG responses to individual Fim2 and Fim3 in sera remaining from a Swedish placebo-controlled efficacy trial that compared a whole-cell vaccine (diphtheria-tetanus-whole-cell pertussis vaccine [DTwP]), a two-component acellular pertussis vaccine (DTaP2), and DTaP5. One month following three doses of the Fim-containing vaccines (DTwP or DTaP5), anti-Fim2 geometric mean IgG concentrations were higher than those for anti-Fim3, with a greater anti-Fim2/anti-Fim3 IgG ratio elicited by DTaP5. We also determined the responses in vaccinated children following an episode of pertussis. Those who received DTaP5 showed a large rise in anti-Fim2 IgG, reflecting the predominant Fim2 serotype at the time. In contrast, those who received DTwP showed an equal rise in anti-Fim2 and anti-Fim3 IgG concentrations, indicating that DTwP may provide a more efficient priming effect for a Fim3 response following contact with B. pertussis. Anti-Fim2 and anti-Fim3 IgG concentrations were also determined in samples from two seroprevalence studies conducted in Sweden in 1997, when no pertussis vaccine was used and Fim2 isolates predominated, and in 2007, when either DTaP2 or DTaP3 without fimbriae was used and Fim3 isolates predominated. Very similar distributions of anti-Fim2 and anti-Fim3 IgG concentrations were obtained in 1997 and 2007, except that anti-Fim3 concentrations in 1997 were lower. This observation, together with the numbers of individuals with both anti-Fim2 and anti-Fim3 IgG concentrations, strongly suggests that B. pertussis expresses both Fim2 and Fim3 during infection.


Modeling of the spread of infection in the interdigital spaces of the manus in limbs from clinically normal dogs.
To determine whether the pattern of extension of modeled infection from the interdigital web spaces in dogs is predictable and whether the distribution differs among initial injury sites. Thawed frozen forelimbs from 23 cadavers of previously healthy adult medium- to large-breed dogs. The manus of each forelimb was evaluated by use of computed tomography (CT) before and after injection of radiopaque blue-staining contrast medium into the interdigital web spaces. Two veterinary radiologists reviewed the CT images and recorded the extent of contrast medium from each site. Each manus was dissected or sectioned transversely after imaging, and the extent of contrast medium accumulation was recorded and compared with locations of CT contrast enhancement. The Fisher exact test was performed to determine whether the pattern of contrast medium extension differed by injection site. Injections made in the interdigital web spaces of the canine manus led to unique and predictable patterns of extension into the surrounding soft tissues. That pattern of extension primarily involved the soft tissues of the digits. In humans, knowledge of common extension patterns from infected soft tissue spaces is used to predict the spread of disease within the hand and develop surgical plans that will minimize patient illness. Identification of the common sites of disease spread from the interdigital web spaces in dogs may help improve surgical planning and treatment for infection in the manus.


Targeted inactivation of the plastid ndhB gene in tobacco results in an enhanced sensitivity of photosynthesis to moderate stomatal closure.
The ndh genes encoding for the subunits of NAD(P)H dehydrogenase complex represent the largest family of plastid genes without a clearly defined function. Tobacco (Nicotiana tabacum) plastid transformants were produced in which the ndhB gene was inactivated by replacing it with a mutant version possessing translational stops in the coding region. Western-blot analysis indicated that no functional NAD(P)H dehydrogenase complex can be assembled in the plastid transformants. Chlorophyll fluorescence measurements showed that dark reduction of the plastoquinone pool by stromal reductants was impaired in ndhB-inactivated plants. Both the phenotype and photosynthetic performance of the plastid transformants was completely normal under favorable conditions. However, an enhanced growth retardation of ndhB-inactivated plants was revealed under humidity stress conditions causing a moderate decline in photosynthesis via stomatal closure. This distinctive phenotype was mimicked under normal humidity by spraying plants with abscisic acid. Measurements of CO(2) fixation demonstrated an enhanced decline in photosynthesis in the mutant plants under humidity stress, which could be restored to wild-type levels by elevating the external CO(2) concentration. These results suggest that the plastid NAD(P)H:plastoquinone oxidoreductase in tobacco performs a significant physiological role by facilitating photosynthesis at moderate CO(2) limitation.


In vivo three-dimensional whole-brain pulsed steady-state chemical exchange saturation transfer at 7 T.
Chemical exchange saturation transfer (CEST) is a technique to indirectly detect pools of exchangeable protons through the water signal. To increase its applicability to human studies, it is needed to develop sensitive pulse sequences for rapidly acquiring whole-organ images while adhering to stringent amplifier duty cycle limitations and specific absorption rate restrictions. In addition, the interfering effects of direct water saturation and conventional magnetization transfer contrast complicate CEST quantification and need to be reduced as much as possible. It is shown that for protons exchanging with rates of less than 50-100 Hz, such as imaged in amide proton transfer experiments, these problems can be addressed by using a three-dimensional steady state pulsed acquisition of limited B(1) strength (≈ 1 μT). Such an approach exploits the fact that the direct water saturation width, magnetization transfer contrast magnitude, and specific absorption rate increase strongly with B(1) , while the size of the CEST effect for such protons depends minimally on B(1) . A short repetition time (65 ms) steady-state sequence consisting of a brief saturation pulse (25 ms) and a segmented echo-planar imaging train allowed acquisition of a three-dimensional whole-brain volume in approximately 11 s per saturation frequency, while remaining well within specific absorption rate and duty cycle limits. Magnetization transfer contrast was strongly reduced, but substantial saturation effects were found at frequencies upfield from water, which still confound the use of magnetization transfer asymmetry analysis. Fortunately, the limited width of the direct water saturation signal could be exploited to fit it with a Lorentzian function allowing CEST quantification. Amide proton transfer effects ranged between 1.5% and 2.5% in selected white and grey matter regions. This power and time-efficient 3D pulsed CEST acquisition scheme should aid endogenous CEST quantification at both high and low fields.


Respiration of the rumen ciliate Dasytricha ruminantium Schuberg.
The endogenous respiration of the rumen ciliate Dasytricha ruminantium maintained under an O2 tension of 2kPa (approximately 0.02 atm) was partially inhibited by KCN (40% inhibition) and NaN3 (58% inhibition). The organisms lack cytochromes, and sensitivity of respiration to KCN, NaN3, chloroquine and quercetin suggest that the operation of flavoprotein-iron-sulphur-mediated electron transport. As in Tritrichomonas foetus, hydrogenosomal respiration can be stimulated by the addition of CoA in the presence of 0.025% Triton X-100; stimulation by ADP was not detected. Stimulation of pyruvate-supported O2 uptake by Pi suggests that acetate is produced via acetyl phosphate.


Vasopressin enhances a calcium current in human ACTH-secreting pituitary adenoma cells.
Arginine vasopressin (AVP) is a potent secretagogue for adrenocorticotropin (ACTH) release from normal corticotropes and from ACTH-secreting pituitary adenoma cells. To explore the mechanism underlying this action, we investigated the effects of AVP on Ca2+-dependent action potentials and Ca2+ currents in cultured human ACTH-containing pituitary tumor cells (hACTH adenoma cells). Pituitary adenoma fragments removed at surgery from two patients with Cushing's disease were dispersed, and the isolated cells were grown in monolayer culture. Most of the cells showed ACTH immunoreactivity that persisted even after as much as 2 months in culture. Current clamp and voltage clamp recordings were carried out using the patch-clamp technique in the whole cell configuration. AVP produced an increase in the amplitude and duration of action potentials in these cells, and substantially enhanced the transient after-hyperpolarization after each spike. Under voltage the transient after-hyperpolarization after each spike. Under voltage clamp, hACTH adenoma cells showed two Ca2+ current components: a low-threshold, rapidly inactivating (T-type) current; and a higher threshold, slowly inactivating (L-type) current. AVP markedly increased the amplitude of the L-type current without affecting the T-type current. These data suggest that AVP may enhance Ca2+ entry associated with action potentials by potentiating the activity of L-type Ca2+ channels. The resulting rise in cytosolic free Ca2+ may be a key link in the process by which AVP stimulates ACTH release in the pituitary.


Nonspecific adherence by Actinobacillus actinomycetemcomitans requires genes widespread in bacteria and archaea.
The gram-negative coccobacillus, Actinobacillus actinomycetemcomitans, is the putative agent for localized juvenile periodontitis, a particularly destructive form of periodontal disease in adolescents. This bacterium has also been isolated from a variety of other infections, notably endocarditis. Fresh clinical isolates of A. actinomycetemcomitans form tenacious biofilms, a property likely to be critical for colonization of teeth and other surfaces. Here we report the identification of a locus of seven genes required for nonspecific adherence of A. actinomycetemcomitans to surfaces. The recently developed transposon IS903phikan was used to isolate mutants of the rough clinical isolate CU1000 that are defective in tight adherence to surfaces (Tad(-)). Unlike wild-type cells, Tad(-) mutant cells adhere poorly to surfaces, fail to form large autoaggregates, and lack long, bundled fibrils. Nucleotide sequencing and genetic complementation analysis revealed a 6.7-kb region of the genome with seven adjacent genes (tadABCDEFG) required for tight adherence. The predicted TadA polypeptide is similar to VirB11, an ATPase involved in macromolecular transport. The predicted amino acid sequences of the other Tad polypeptides indicate membrane localization but no obvious functions. We suggest that the tad genes are involved in secretion of factors required for tight adherence of A. actinomycetemcomitans. Remarkably, complete and highly conserved tad gene clusters are present in the genomes of the bubonic plague bacillus Yersinia pestis and the human and animal pathogen Pasteurella multocida. Partial tad loci also occur in strikingly diverse Bacteria and Archaea. Our results show that the tad genes are required for tight adherence of A. actinomycetemcomitans to surfaces and are therefore likely to be essential for colonization and pathogenesis. The occurrence of similar genes in a wide array of microorganisms indicates that they have important functions. We propose that tad-like genes have a significant role in microbial colonization.


[Peracute suffocation emergency: what should be done?].
Peracute obstructive dyspnea with imminent asphyctic death is a dramatic emergency. It is characterized by easily audible inspiratory stridor caused by severe stenosis. Several causes in the pharynx and larynx may be responsible. Pharmacotherapy is hardly effective, an intubation extremely difficult in extremis. In contrast emergency coniotomy offers a real chance to reopen the airways even to the surgically inexperienced. The technique for this intervention is described.


Needle EMG of the tongue: motor unit action potential versus peak ratio analysis in limb and bulbar onset amyotrophic lateral sclerosis.
To find out if conventional and automatic needle EMG of the tongue can be helpful in the diagnosis and differentiation of limb and bulbar onset amyotrophic lateral sclerosis. Motor unit action potential (MUAP) analysis and peak ratio interference pattern analysis were performed in the right genioglossus muscle of 30 healthy subjects aged 30-81 years, 10 patients aged 49-73 years with limb onset amyotrophic lateral sclerosis, and eight patients aged 52-75 years with bulbar onset amyotrophic lateral sclerosis. Electrical activity was sampled via standard concentric needle electrodes with a commercially available EMG recorder. Normal mean (2SD) MUAP duration was 6.6 (1.5) ms. Normal mean (2SD) MUAP amplitude was 224 (97.4) microV. Normal mean (2SD) peak ratio (PR), turns/second (T/s), amplitude/turn (A/T), and time intervals (TI1, TI2, TI3) were 1.68 (0.56), 732 (303.9), 446 (180.3) microV, 2.62 (0.34), 2.31 (0.14), and 1.01 (0.50) respectively. Mean MUAP duration and amplitude were significantly increased in limb onset (P=0.0001 and P=0.013) and bulbar onset amyotrophic lateral sclerosis (P=0.0001 and P=0.017). Peak ratio indices stayed unchanged in limb onset amyotrophic lateral sclerosis but were significantly decreased (PR, T/s, A/T, TI1, and TI2) or increased (TI3) in bulbar onset disease. The sensitivity of the MUAP analysis was 70% in limb and 75% in bulbar onset amyotrophic lateral sclerosis. The sensitivity of the peak ratio interference pattern analysis was 20% in limb and 100% in bulbar onset amyotrophic lateral sclerosis. Subclinical involvement of the tongue was found in 20% of the patients with limb onset amyotrophic lateral sclerosis and could be more accurately assessed with MUAP analysis than with automatic EMG. Both conventional and automatic needle EMG of the tongue are valuable electrophysiological devices to assess the clinical and subclinical involvement of the tongue in patients with limb and bulbar onset amyotrophic lateral sclerosis.


Heart rate variability and ischaemia in patients with coronary heart disease and stable angina pectoris; influence of drug therapy and prognostic value. TIBBS Investigators Group. Total Ischemic Burden Bisoprolol Study.
Determination of the influence of therapy with bisoprolol and nifedipine on the heart rate variability of patients from the Total Ischemic Burden Bisoprolol Study and examination of the prognostic value. Four hundred and twenty-two patients with stable angina were included. The heart rate variability was determined over a period of 24 h. Parameters determined: standard deviation of the mean of all corrected RR intervals, standard deviation of all 5 min mean cycle lengths, square root of the mean of the squared differences of successive corrected RR intervals. Nifedipine reduced the mean values of all heart rate variability parameters tested. Square root of the mean of the square differences of successive corrected RR intervals increased under bisoprolol. Standard deviation of the mean of all corrected RR intervals and standard deviation of all 5 min mean cycle lengths increased from low baseline values and declined from higher baseline values. The increase in heart rate variability under therapy was accompanied by a tendency towards a better prognosis. Patients with an increase in heart rate variability and simultaneous complete suppression of ischaemia under therapy displayed no serious events in the course of one year. The increase in the heart rate variability, which can be regarded as prognostically favourable, was predominantly observed under bisoprolol. The parameter constellation of an increase in heart rate variability and complete ischaemia suppression on the 48-h Holter ECG was associated with the greatest benefit.


Suspension laryngoscopy for the thoracic surgeon: when and how to use it.
Suspension laryngoscopy is one of the most common otolaryngological procedures for the diagnosis and surgical approach to the larynx. However, most thoracic surgeons are not familiar with the procedure and seldom use it. The indications for its use are similar to those for that of rigid bronchoscopy (dilatation, endoprosthesis insertion, and tumor resection). It can be performed in children and adults. Suspension laryngoscopy is an alternative when rigid bronchoscopy is unavailable and is therefore a viable option for use at smaller facilities. In this communication, we describe the technique and the applications of suspension laryngoscopy in thoracic surgery.


Zero Liquid Discharge approach in plating industry: treatment of degreasing effluents by electrocoagulation and anodic oxidation.
Degreasing waste effluents issued from a surface treatment plant were treated by electrochemical techniques in an attempt to reduce COD so that clean water can be returned to the rinse bath. Electrocoagulation, both with iron and aluminium anodes, and anodic oxidation with boron doped diamond (BDD) anodes were tested. In the electrocoagulation tests, the nature of the anodes did not impact significantly the reduction of COD. Electrocoagulation showed good COD removal rates, superior to 80%, but it was not able to reduce COD down to low levels. Anodic oxidation was able to reduce COD down to discharge limits; the oxidation efficiency was superior to 50%. Economical calculations show that anodic oxidation is best used as a polishing step after electrocoagulation. The bulk of the COD would be reduced by electrocoagulation and, then, anodic oxidation would reduce COD below discharge limits. The maximum treatable flow is somewhat hindered by the small sizes of current BDD installation but it would reach 600 m(3)/year if anodic oxidation is coupled with electrocoagulation, the operational cost being 2.90 Euros /m(3).


The EQ-5D-5L valuation study in Korea.
This study aimed to estimate Korean preference weights for EQ-5D-5L based on values elicited from Korean population by applying the EuroQol Valuation Technology (EQ-VT) program and the standard protocol by the EuroQol Group. The multistage quota sampling method was used to recruit 1085 subjects from the general population in Korea. Each respondent valuated 10 health states using the composite time trade-off (cTTO) and 7 health states using discrete choice experiment. The EQ-VT program was developed by the EuroQol Group and translated into Korean with the Korean research team. Computer-assisted, face-to-face interviews were conducted. A range of predictive models were explored using cTTO. The most appropriate model was determined after assessing goodness of fit, logical consistency, and parsimony. Of 3206 contacted, 1085 subjects completed interviews (33.8 %) and 1080 were used for modeling. A model with dummy variables for each level of severity and dimension and a term that picked up whether any dimension in the state was at level 4 or 5 was selected as the best predictive model. All coefficients of the final model were statistically significant and logically consistent. In addition, it was parsimonious. This model had mean absolute error of 0.027, and the absolute error for all 86 health states was below 0.1. The final model built in this study appears to predict the utilities of the states which were valuated directly. This model could be used to interpolate quality weights for all EQ-5D-5L health states.


The effects of training with high-speed interval running on muscle performance are modulated by slope.
We examined changes in selected muscle performance parameters after 8 weeks of interval training using two opposite running inclinations. We hypothesized that the uphill training will affect endurance muscle performance outcomes, whereas the downhill training will affect power muscle performance outcomes. Fourteen physically active volunteers were randomly assigned into either the Uphill group (UG; n = 7; uphill interval running at +10% incline) or the Downhill group (DG; n = 7; downhill interval running at -10% incline) and completed 16 training sessions. Each session consisted of ten 30 s treadmill runs at 90% of maximum aerobic speed (MAS) with a work to rest ratio of 1:2. Vertical jump performance, isometric (MVC) and isokinetic torque of knee extensors and flexors, and fatigue of knee extensors were evaluated pre and post-training. Moreover, body composition (via bioimpedance) and vastus lateralis muscle architecture (via ultrasonography) were assessed pre and post-training. Relative lean tissue mass, relative fat mass, and squat jump (cm) significantly (p < .05) changed from baseline values by +4.5 ± 4.0%, -11.5 ± 9.6%, and +9.5 ± 11.7%, respectively, only in the DG. Similarly, DG improved absolute values of knee extension rate of torque development and impulse (p < .05), whereas knee flexion peak torque angle significantly decreased in both groups (p < .05). On the other hand, the UG increased the number of repetitions achieved during the fatigue protocol and total work by 21.2 ± 32.6% and 13.8 ± 21.2%, respectively (p < .05). No differences were found between groups in muscle architecture. Introducing variations in slope during HIIT could be used to induce specific improvements toward muscle endurance or power performance characteristics.


Susceptibility Testing of Fungi to Antifungal Drugs.
Susceptibility testing of fungi against antifungal drugs commonly used for therapy is a key component of the care of patients with invasive fungal infections. Antifungal susceptibility testing (AFST) has progressed in recent decades to finally become standardized and available as both Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) reference methods and in commercial manual/automated phenotypic methods. In clinical practice, the Sensititre YeastOne and Etest methods are widely used for AFST, particularly for sterile site isolates of Candida. Nevertheless, AFST is moving toward new phenotypic methods, such as matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), that are capable of providing rapid, and potentially more actionable, results for the treating clinician. Our objective is to summarize updated data on phenotypic methods for AFST of Candida and Aspergillus species and to assess their significance in view of opposing, but emerging, molecular genotypic methods.


Local immune responses of the Chinese water buffalo, Bubalus bubalis, against Schistosoma japonicum larvae: crucial insights for vaccine design.
Asian schistosomiasis is a zoonotic parasitic disease infecting up to a million people and threatening tens of millions more. Control of this disease is hindered by the animal reservoirs of the parasite, in particular the water buffalo (Bubalus bubalis), which is responsible for significant levels of human transmission. A transmission-blocking vaccine administered to buffaloes is a realistic option which would aid in the control of schistosomiasis. This will however require a better understanding of the immunobiology of schistosomiasis in naturally exposed buffaloes, particularly the immune response to migrating schistosome larvae, which are the likely targets of an anti-schistosome vaccine. To address this need we investigated the immune response at the major sites of larval migration, the skin and the lungs, in previously exposed and re-challenged water buffaloes. In the skin, a strong allergic-type inflammatory response occurred, characterised by leukocyte and eosinophil infiltration including the formation of granulocytic abscesses. Additionally at the local skin site, interleukin-5 transcript levels were elevated, while interleukin-10 levels decreased. In the skin-draining lymph node (LN) a predominant type-2 profile was seen in stimulated cells, while in contrast a type-1 profile was detected in the lung draining LN, and these responses occurred consecutively, reflecting the timing of parasite migration. The intense type-2 immune response at the site of cercarial penetration is significantly different to that seen in naive and permissive animal models such as mice, and suggests a possible mechanism for immunity. Preliminary data also suggest a reduced and delayed immune response occurred in buffaloes given high cercarial challenge doses compared with moderate infections, particularly in the skin. This study offers a deeper understanding into the immunobiology of schistosomiasis in a natural host, which may aid in the future design of more effective vaccines.


Iterative linear focal-plane wavefront correction.
We propose an efficient approximation to the nonlinear phase diversity (PD) method for wavefront reconstruction and correction from intensity measurements with potential of being used in real-time applications. The new iterative linear phase diversity (ILPD) method assumes that the residual phase aberration is small and makes use of a first-order Taylor expansion of the point spread function (PSF), which allows for arbitrary (large) diversities in order to optimize the phase retrieval. For static disturbances, at each step, the residual phase aberration is estimated based on one defocused image by solving a linear least squares problem, and compensated for with a deformable mirror. Due to the fact that the linear approximation does not have to be updated with each correction step, the computational complexity of the method is reduced to that of a matrix-vector multiplication. The convergence of the ILPD correction steps has been investigated and numerically verified. The comparative study that we make demonstrates the improved performance in computational time with no decrease in accuracy with respect to existing methods that also linearize the PSF.


What governs governance, and how does it evolve? The sociology of governance-in-action.
Governance addresses a wide range of issues including social, economic and political continuity, security and integrity, individual and collective safety and the liberty and rights to self-actualization of citizens. Questions to be answered include how governance can be achieved and sustained within a social context imbued with cultural values and in which power is distributed unevenly and dynamically, and how governance impacts on individuals and institutions. Drawing on Gramscian notions of hegemony and consent, and recent political science literatures on regulation and meta-regulation, this paper develops a sociological model of governance that emphasizes a dynamic and responsive governance in action. Empirical data from a study of pharmaceutical governance is used to show how multiple institutions and actors are involved in sustaining effective governance. The model addresses issues of how governance is sustained in the face of change, why governance of practices varies from setting to setting, and how governance is achieved without legislation.


Relaxation and related therapies for people with multiple sclerosis (MS): A systematic review.
To establish the effectiveness of relaxation and related therapies in treating Multiple Sclerosis related symptoms and sequelae. PsycINFO, PubMed, Embase, CINAHL, ProQuest Dissertations and Theses Global databases were searched. We included studies from database inception until 31 December 2021 involving adult participants diagnosed with multiple sclerosis or disseminated sclerosis, which featured quantitative data regarding the impact of relaxation interventions on multiple sclerosis-related symptoms and sequelae. Studies which examined multi-modal therapies - relaxation delivered in combination with non-relaxation interventions - were excluded. Risk of bias was assessed using the Revised Risk of Bias tool for randomised trials - ROB2, Risk of Bias in Non-Randomised Studies of Interventions ROBINS-I), and within and between-group effects were calculated (Hedges' g). Twenty-eight studies met inclusion criteria. Twenty-three of these were randomised controlled trials, with 1246 total participants. This review reports on this data, with non-randomised study data reported in supplemental material. Post -intervention relaxation was associated with medium to large effect-size improvement for depression, anxiety, stress and fatigue. The effects of relaxation were superior to wait-list or no treatment control conditions; however, comparisons with established psychological or physical therapies were mixed. Individual studies reported sustained effects (≤ 6 months) with relaxation for stress, pain and quality of life. Most studies were rated as having a high/serious risk of bias. There is emerging evidence that relaxation therapies can improve outcomes for persons with multiple sclerosis. Given the high risk of bias found for included studies, stronger conclusions cannot be drawn.


Coexistence of Epstein-Barr virus and Parvovirus B19 in tonsillar tissue samples: quantitative measurement by real-time PCR.
In this study, we aimed to investigate the presence and copy number of six different viruses in tonsillar tissue samples removed surgically because of chronic recurrent tonsillitis or chronic obstructive tonsillar hypertrophy. In total, 56 tissue samples (tonsillar core) collected from 44 children and 12 adults were included in this study. The presence of viruses was investigated using a new TaqMan-based quantitative real-time PCR assay. Of the 56 tissue samples, 67.9% (38/56) were positive for at least one of the six viruses. Epstein-Barr virus was the most frequently detected virus, being found in 53.6% (30/56), followed by human Parvovirus B19 21.4% (12/56), human adenovirus 12.5% (7/56), human Cytomegalovirus 5.4% (3/56), BK polyomavirus 1.8% (1/56), and Herpes simplex virus 1.8% (1/56). Precancerous or cancerous changes were not detected in the tonsillar tissue samples by pathologic examination, whereas lymphoid hyperplasia was observed in 24 patients. In contrast to other viruses, B19 virus was present in high copy number in tonsillar tissues. The rates of EBV and B19 virus with high copy number (>500.000 copies/ml) were higher in children than in adults, and a positive relationship was also found between the presence of EBV and the presence of B19 virus with high copy number (P=0.037). It is previously reported that some viral agents are associated with different chronic tonsillar pathologies. In the present study, the presence of B19 virus in tonsillar core samples was investigated quantitatively for the first time, and our data suggests that EBV infections could be associated with B19 virus infections or could facilitate B19 virus replication. However, further detailed studies are needed to clarify this observation.


The neural system underlying Chinese logograph reading.
Written Chinese as logographic script differs notably from alphabets such as English in visual form, orthography, phonology, and semantics. Thus, research on the Chinese language is important to advance our understanding of the universality and particularity of the organization of language systems in the brain. In this study, we examine the neural systems associated with logographic reading using functional magnetic resonance imaging. Two experimental tasks were devised, one based on semantic decision and the other on homophone decision. Compared to the fixation baseline, peak activations resulting from semantic as well as homophony decisions were localized in the left middle frontal gyrus (BA 9). Left inferior frontal cortex also mediated Chinese processing. In addition, more right hemisphere cortical regions (i.e., BAs 47/45, 7, 40/39, and the right visual system) were involved in reading Chinese relative to reading English. This is attributed to the square shape of the logograph which requires an elaborated analysis of the spatial information and locations of various strokes comprising the logographic character. We suggest that the left middle frontal area (BA 9) coordinates and integrates the intensive visuospatial analysis demanded by logographs' square configuration and the semantic (or phonological) analysis required by the present experimental tasks. Our study has implicated brain regions common to both logographic and alphabetic languages as well as brain regions specialized in processing logographs.


On gel electrophoresis of dielectric charged particles with hydrophobic surface: A combined theoretical and numerical study.
A theoretical study on the gel electrophoresis of a charged particle incorporating the effects of dielectric polarization and surface hydrophobicity at the particle-liquid interface is made. A simplified model based on the weak applied field and low charge density assumption is also presented and compared with the full numerical model for a nonpolarizable particle to elucidate the nonlinear effects such as double layer polarization and relaxation as well as surface conduction. The main motivation of this study is to analyze the electrophoresis of the surface functionalized nanoparticle with tunable hydrophobicity or charged fluid drop in gel medium by considering the electrokinetic effects and hydrodynamic interactions between the particle and the gel medium. An effective medium approach, in which the transport in the electrolyte-saturated hydrogel medium is governed by the Brinkman equation, is adopted in the present analysis. The governing electrokinetic equations based on the conservation principles are solved numerically. The Navier-slip boundary condition along with the continuity condition of dielectric displacement are imposed on the surface of the hydrophobic polarizable particle. The impact of the slip length on the electrophoresis is profound for a thinner Debye layer, however, surface conduction effect also becomes significant for a hydrophobic particle. Impact of hydrophobicity and relaxation effects are higher for a larger particle. Dielectric polarization creates a reduction in its electrophoretic propulsion and has negligible impact at the thinner Debye length as well as lower gel screening length.


Selection of Fusion Levels in Adolescent Idiopathic Scoliosis.
PURPOSE OF REVIEW: Posterior spinal fusion (PSF) is the preferred treatment for adolescent idiopathic scoliosis (AIS) patients with surgical range curves. Selection of the proper upper and lower instrumented vertebrae (UIV and LIV) is essential in curve correction and achieving a successful outcome, while preventing short and long-term complications. RECENT FINDINGS: The literature lacks high-level evidence, especially on outcomes of modern surgical techniques. However, evidence seems to show that a great majority of AIS patients have excellent clinical and functional long-term outcomes after PSF. We have reviewed the evidence and provided our level selection recommendations, which should be weighed against the body of evidence on the topic when selecting fusion levels in AIS.


[Effect of intravenous administration of nitroglycerin on the indicators of central hemodynamics in patients with acute myocardial infarction].
The effect of intravenous nitroglycerin administration on central hemodynamics criteria (end diastolic pressure in the pulmonary artery and cardiac output) was analysed in a controlled study of 60 patients with acute myocardial infarction. Nitroglycerin-induced changes in the parameters were shown to be related to the hemodynamic state at baseline, with the improvement in left ventricular functional state being significant in the presence of left ventricular insufficiency and the changes minimal in the presence of normal baseline parameters. Thus, nitroglycerin can be used in patients with hemodynamic alterations without a detrimental effect.


RNA virus genetic robustness: possible causes and some consequences.
In general terms, robustness is the capacity of biological systems to function in spite of genetic or environmental perturbations. The small and compacted genomes and high mutation rates of RNA viruses, as well as the ever-changing environments wherein they replicate, create the conditions for robustness to be advantageous. In this review, I will enumerate possible mechanisms by which viral populations may acquire robustness, distinguishing between mechanisms that are inherent to virus replication and population dynamics and those that result from the interaction with host factors. Then, I will move to review some evidences that RNA virus populations are robust indeed. Finally, I will comment on the implications of robustness for virus evolvability, the emergence of new viruses and the efficiency of lethal mutagenesis as an antiviral strategy.


Growth asymmetry precedes differential auxin response during apical hook initiation in Arabidopsis.
The development of a hook-like structure at the apical part of the soil-emerging organs has fascinated botanists for centuries, but how it is initiated remains unclear. Here, we demonstrate with high-throughput infrared imaging and 2-D clinostat treatment that, when gravity-induced root bending is absent, apical hook formation still takes place. In such scenarios, hook formation begins with a de novo growth asymmetry at the apical part of a straightly elongating hypocotyl. Remarkably, such de novo asymmetric growth, but not the following hook enlargement, precedes the establishment of a detectable auxin response asymmetry, and is largely independent of auxin biosynthesis, transport and signaling. Moreover, we found that functional cortical microtubule array is essential for the following enlargement of hook curvature. When microtubule array was disrupted by oryzalin, the polar localization of PIN proteins and the formation of an auxin maximum became impaired at the to-be-hook region. Taken together, we propose a more comprehensive model for apical hook initiation, in which the microtubule-dependent polar localization of PINs may mediate the instruction of growth asymmetry that is either stochastically taking place, induced by gravitropic response, or both, to generate a significant auxin gradient that drives the full development of the apical hook.


Damage control resuscitation: a sensible approach to the exsanguinating surgical patient.
The current wars in Iraq and Afghanistan have resulted in the highest rates of combat casualties experienced by the U.S. military since the Vietnam conflict. These casualties suffer wounds that have no common civilian equivalent and more frequently require massive transfusion (greater than 10 units of packed red blood cells [PRBCs] in less than 24 hrs) than civilian injured. Military surgeons have found that traditional approaches to resuscitation, particularly in terms of the ratio of blood products to each other and the timing of these products, often fail to effectively treat the coagulopathy that is present on arrival in these casualties. This observation has been concurrently noted in the civilian trauma literature. These experiences have ignited interest in an alternative approach to the resuscitation of these most grievously injured patients. This approach includes the use of permissive hypotension; the prevention and aggressive treatment of hypothermia with both passive and active warming measures; the temporization of acidosis with use of exogenous buffer agents; the immediate use of thawed plasma in ratios approaching 1:1 with PRBCs; the early use of platelets, often given well before 10 units of PRBCs have been transfused; the early use of recombinant Factor VIIa; and, in military settings, the use of fresh whole blood as a primary resuscitation fluid. This strategy has been called "damage control resuscitation" to emphasize its pairing with damage control surgical techniques. Review of the published support for this strategy reveals that additional trials are needed to study and optimize these techniques.


Investigation of Narrow-band NIR LED Spectral Response Towards a Non-invasive Glucose Sensor.
Spectroscopy is utilised extensively in medical sensing technology. Typically, hand-held spectroscopy equipment uses miniature narrow-band light emitting diodes (LEDs) and photodiodes to emit and detect light, respectively. Photodiodes typically absorb light across a wide spectra so measurements can be corrupted by surrounding light. LEDs in the visible spectrum have a narrower spectral response and can be used in place of a traditional photodiode. However, the absorption characteristics of near infrared (NIR) spectrum LEDs is unknown. A discrete, low-cost spectrophotometer was designed to assess spectral response for 8 narrow band NIR LEDs. The normalised and raw spectral response determined the optimum detector for 1050 nm - 1300 nm is the 1450 nm LED, and the optimum detector for 1450 nm - 1650 nm emissions is the 1650 nm LED.Clinical relevance - Understanding the spectral response of narrow-band LEDs in the NIR spectrum will aid development of NIR hand-held spectroscopy medical devices.


Neutralizing immune responses induced by oligomeric H5N1-hemagglutinins from plants.
Plant-based transient expression is an alternative platform to produce hemagglutinin-based subunit vaccines. This production system provides not only fast and effective response in the context of a pandemic but also enables the supply of big volume vaccines at low cost. Crude plant extracts containing influenza hemagglutinin are considered to use as vaccine sources because of avoidance of related purification steps resulting in low cost production allowing veterinary applications. Highly immunogenic influenza hemagglutinins are urgently required to meet these pre-conditions. Here, we present a new and innovative way to generate functional H5 oligomers from avian flu hemagglutinin in planta by the specific interaction of S·Tag and S·Protein. A S·Tag was fused to H5 trimers and this construct was transiently co-expressed in planta with S·Protein-TPs which was multimerized by disulfide bonds via cysteine residues in tailpiece sequences (TP) of IgM antibody. Multimerized S·Protein-TPs serve as bridges/molecular docks to combine S·Tag-fused hemagglutinin trimers to form very large hemagglutinin H5 oligomers. H5 oligomers in the plant crude extract were highly active in hemagglutination resulting in high titers. Immunization of mice with two doses of plant crude extracts containing H5 oligomers after storage for 1 week at 4 °C caused strong immune responses and induced neutralizing specific humoral immune responses in mice. These results allow for the development of cheap influenza vaccines for veterinary application in future.


2-Hydroxyglutarate destabilizes chromatin regulatory landscape and lineage fidelity to promote cellular heterogeneity.
The epigenome delineates lineage-specific transcriptional programs and restricts cell plasticity to prevent non-physiological cell fate transitions. Although cell diversification fosters tumor evolution and therapy resistance, upstream mechanisms that regulate the stability and plasticity of the cancer epigenome remain elusive. Here we show that 2-hydroxyglutarate (2HG) not only suppresses DNA repair but also mediates the high-plasticity chromatin landscape. A combination of single-cell epigenomics and multi-omics approaches demonstrates that 2HG disarranges otherwise well-preserved stable nucleosome positioning and promotes cell-to-cell variability. 2HG induces loss of motif accessibility to the luminal-defining transcriptional factors FOXA1, FOXP1, and GATA3 and a shift from luminal to basal-like gene expression. Breast tumors with high 2HG exhibit enhanced heterogeneity with undifferentiated epigenomic signatures linked to adverse prognosis. Further, ascorbate-2-phosphate (A2P) eradicates heterogeneity and impairs growth of high 2HG-producing breast cancer cells. These findings suggest 2HG as a key determinant of cancer plasticity and provide a rational strategy to counteract tumor cell evolution.


Therapeutic benefits of qigong exercises in combination with drugs.
This article reviews clinical studies from the Qigong Bibliographic Database, developed by the Qigong Institute, a nonprofit organization. This database was started in 1994 and holds approximately 1300 references going back to 1986, covering medical applications, scientific, and experimental studies on qigong from China, the United States, and Europe. Records in English have been compiled from International Qigong conferences and seminars, scientific journals, magazines, dissertations, MEDLINE, and other databases. The therapeutic role of qigong exercises combined with drugs is reported for three medical conditions that require drug therapy for health maintenance: hypertension, respiratory disease, and cancer. In these studies, drugs were administered to all patients who were divided into two groups, a group that practiced qigong exercises and a control group that did not. Taken together, these studies suggest that practicing qigong exercises may favorably affect many functions of the body, permit reduction of the dosage of drugs required for health maintenance, and provide greater health benefits than the use of drug therapy alone. For hypertensive patients, combining qigong practice with drug therapy for hypertensive patients resulted in reduced incidence of stroke and mortality and reduced dosage of drugs required for blood pressure maintenance. For asthma patients, the combination therapy permitted reduction in drug dosage, the need for sick leave, duration of hospitalization, and costs of therapy. For cancer patients, the combination therapy reduced the side effects of cancer therapy. Also reported is a study showing that the practice of qigong helps to rehabilitate drug addicts. The reported studies do not necessarily measure up to the strict protocols required for randomized controlled clinical trials.


Incidence and prognostic impact of amenorrhea during adjuvant therapy in high-risk premenopausal breast cancer: analysis of a National Cancer Institute of Canada Clinical Trials Group Study--NCIC CTG MA.5.
To investigate the therapeutic impact of chemotherapy-induced amenorrhea in premenopausal patients with breast cancer. We conducted a retrospective cohort study of a National Cancer Institute of Canada Clinical Trials Group phase III trial involving premenopausal patients randomized to receive cyclophosphamide, methotrexate, and fluorouracil (CMF), versus intensive cyclophosphamide, epirubicin, and fluorouracil (CEF). The objectives of our study were to describe the incidence of amenorrhea at 6 and 12 months post-random assignment and to determine the association of amenorrhea with relapse-free and overall survival. Data on 442 patients were used in our analyses. Despite the higher cumulative dose of cyclophosphamide in the CMF treatment arm, at 6 months post-random assignment, the rate of amenorrhea was higher in the CEF group (relative risk, 1.2; 95% CI, 1.0 to 1.3), with no difference at 12 months. In the receptor-positive subgroup, 6-month amenorrhea rates were not associated with prognosis. In contrast, amenorrhea at 12 months was significantly associated with relapse-free survival (hazard ratio, 0.51; 95% CI, 0.32 to 0.82; P = .005) and overall survival (hazard ratio, 0.40; 95% CI, 0.22 to 0.72; P = .002). Late chemotherapy-induced amenorrhea seems to be associated with improved outcome in patients with premenopausal, receptor-positive breast cancer.


Temporal trends in the relative survival among patients diagnosed with ovarian cancer in Canada 1992-2005: a population-based study.
Ovarian cancer affects about 2600 women annually in Canada and about 1750 are expected to die from this disease. We estimated the trends in the relative survival ratio for patients diagnosed with epithelial invasive ovarian cancer in Canadian population between 1992 and 2005. A flexible parametric model was used to estimate the relative survival ratio. Relative survival ratio is defined as the observed survival among cancer patients divided by the expected survival in the general population. We incorporated age group, histology of tumour, and region of patient residence at the time of diagnosis into a model to predict two- and five-year relative survival ratios based on the year of diagnosis where the effect of each variable was adjusted for the effects of the other variables. A restricted cubic splines with five knots was used to include year of diagnosis in the model. In total 7771 patients diagnosed with epithelial invasive ovarian cancer were included in this analysis with the mean age of 59.6 (SD=13.5) years at the time of diagnosis. About 75% of the patients were 50 years and older at the time of diagnosis and relative survival ratio substantially decreased with age. The tumour type was serous for 43% of cases followed by endometrioid (30.5%), clear cell (14.5%), mucinous (9.5%), and transitional cell (2.5%). The same pattern was observed for all regions although with some variation in the proportions. The worst survival observed for serous tumours. About 50% of the cases were diagnosed in Ontario. This is the first report that compares relative survival ratio for epithelial invasive ovarian cancer among geographic regions of Canada where a higher relative survival ratio was observed in Ontario compared to the other regions. The relative survival decreased with age and showed geographic variation. The work indicates that advances in management of women with ovarian cancer have improved two- and five-year relative survival ratios.


Ectopic expression of insulin in a type 1 diabetic rat model by injection of manipulated mesenchymal stem cells with an insulin construct driven by a glucose-sensitive promoter in the port vein.
The treatment of type 1 diabetes through islet cell transplantation is a complex process, facing challenges such as allograft rejections and a limited supply of donors. One potential solution is to utilize the liver as an alternative for natural insulin production, as hepatocytes can secrete proteins and respond to glucose levels. Recent research has shown promising results in using mesenchymal stem cells as a potential cure for diabetes. The study utilized a diabetic rat model, confirmed through blood sugar measurement. A plasmid vector was designed with specific genetic components, synthesized by biotech company, and then Inserted vector into a plasmid with resistance genes and bacterial origin. Bone marrow-derived mesenchymal stem cells (BM-MSCs) were cultured and transfected with the plasmid using Lipofectamine 3000. Polymerase chain reaction was employed to confirm successful transfection using specific primers. For the animal study, 30 male Wistar rats were divided into six groups, each comprising five rats. The control group did not receive any treatment, while the second group received MSCs via Portal Vein Injection. The third group received MSCs transfected with a specific construct via Portal Vein Injection. The fourth group was induced to develop diabetes through streptozotocin (STZ) injection, the fifth group developed diabetes and received untransfected MSCs via Portal Vein Injection, and the sixth group received MSCs transfected with the specific construct via Portal Vein Injection. To manage Pain, appropriate pain control was administered to the rats for 3 days after the surgery. Fixed liver tissues obtained from the euthanized rats were utilized for immunohistochemistry. In this study, immunohistochemical techniques were used to examine insulin expression in different groups of rats. The control groups showed high levels of insulin expression, while the diabetic groups exhibited lower expression. However, there was a significant difference between the diabetic groups treated with MSC and transgenic MSC cells. All groups had similar baseline glucose levels, but the diabetic groups showed a significant increase after STZ injection, whereas the control and MSC groups did not. Postintervention, both the control and MSC groups had similar glucose levels to the post-STZ levels. However, diabetes-induced groups experienced a significant decrease in glucose levels, with the transfected MSCs showing a greater decrease than the untransfected MSCs. The study suggested that treatment with MSCs, especially transfected ones, can effectively reduce glucose levels in rats with diabetes. In this research, rat BM-MSCs were utilized to create insulin-producing mesenchymal cells with glucose-sensitive insulin expression. The cells were transferred to the liver of diabetic rats via portal vein injection, leading to an increase in insulin expression. This study proposes a novel approach for cell therapy and delivery in the treatment of type 1 diabetes.


Lyme disease.
Within the last decade, Lyme borreliosis has emerged as a complex new infection whose distribution is worldwide. The disorder is caused by a recently recognized spirochete, B. burgdorferi, transmitted by ticks of the I. ricinus complex. Certain species of mice are critical in the life cycle of the spirochete, and deer appear to be crucial to the tick. Although the disorder's basic outlines are similar everywhere, there are regional variations in the causative spirochete, animal hosts, and clinical manifestations of the illness. In the United States, Lyme disease commonly begins in summer with a characteristic skin lesion, erythema migrans, accompanied by flu-like or meningitis-like symptoms. Weeks or months later, the patients may have neurologic or cardiac abnormalities, migratory musculoskeletal pain, or arthritis, and more than a year after onset, some patients have chronic joint, skin, or neurologic abnormalities. After the first several weeks of infection, almost all patients have a positive antibody response to the spirochete, and serologic determinations are currently the most practical laboratory aid in diagnosis. Treatment with appropriate antibiotics is usually curative, but longer courses of therapy are often needed later in the illness, and some patients may not respond.


Regulatory consequences of gene translocation in bacteria.
Gene translocations play an important role in the plasticity and evolution of bacterial genomes. In this study, we investigated the impact on gene regulation of three genome organizational features that can be altered by translocations: (i) chromosome position; (ii) gene orientation; and (iii) the distance between a target gene and its transcription factor gene ('target-TF distance'). Specifically, we quantified the effect of these features on constitutive expression, transcription factor binding and/or gene expression noise using a synthetic network in Escherichia coli composed of a transcription factor (LacI repressor) and its target gene (yfp). Here we show that gene regulation is generally robust to changes in chromosome position, gene orientation and target-TF distance. The only demonstrable effect was that chromosome position alters constitutive expression, due to changes in gene copy number and local sequence effects, and that this determines maximum and minimum expression levels. The results were incorporated into a mathematical model which was used to quantitatively predict the responses of a simple gene network to gene translocations; the predictions were confirmed experimentally. In summary, gene translocation can modulate constitutive gene expression levels due to changes in chromosome position but it has minimal impact on other facets of gene regulation.


Assessment of advanced glycated end product accumulation in skin using auto fluorescence multispectral imaging.
Several studies have shown that advanced glycation end products (AGE) play a role in both the microvascular and macrovascular complications of diabetes and are closely linked to inflammation and atherosclerosis. AGEs accumulate in skin and can be detected using their auto fluorescence (AF). A significant correlation exists between AGE AF and the levels of AGEs as obtained from skin biopsies. A commercial device, the AGE Reader, has become available to assess skin AF for clinical purposes but, while displaying promising results, it is limited to single-point measurements performed in contact to skin tissue. Furthermore, in vivo imaging of AGE accumulation is virtually unexplored. We proposed a non-invasive, contact-less novel technique for quantifying fluorescent AGE deposits in skin tissue using a multispectral imaging camera setup (MSI) during ultraviolet (UV) exposure. Imaging involved applying a region-of-interest mask, avoiding specular reflections and a simple calibration. Results of a study conducted on 16 subjects with skin types ranging from fair to deeply pigmented skin, showed that AGE measured with MSI in forearm skin was significantly correlated with the AGE reference method (AGE Reader on forearm skin, R=0.68, p=0.005). AGE measured in facial skin was borderline significantly related to AGE Reader on forearm skin (R=0.47, p=0.078). These results support the use of the technique in devices for non-touch measurement of AGE content in either facial or forearm skin tissue over time.


Health workforce planning: which countries include nurse practitioners and physician assistants and to what effect?
An increasing number of countries are introducing new health professions, such as Nurse Practitioners (NPs) and Physician Assistants (PAs). There is however limited evidence, on whether these new professions are included in countries' workforce planning. A cross-country comparison of workforce planning methods. Countries with NPs and/or PAs were identified, workforce planning projections reviewed and differences in outcomes were analysed, based on a review of workforce planning models and a scoping review. Data on multi-professional (physicians/NPs/PAs) vs. physician-only models were extracted and compared descriptively. Analysis of policy implications was based on policy documents and grey literature. Of eight countries with NPs/PAs, three (Canada, the Netherlands, United States) included these professions in their workforce planning. In Canada, NPs were partially included in Ontario's needs-based projection, yet only as one parameter to enhance efficiency. In the United States and the Netherlands, NPs/PAs were covered as one of several scenarios. Compared with physician-only models, multi-professional models resulted in lower physician manpower projections, primarily in primary care. A weakness of the multi-professional models was the accuracy of data on substitution. Impacts on policy were limited, except for the Netherlands. Few countries have integrated NPs/PAs into workforce planning. Yet, those with multi-professional models reveal considerable differences in projected workforce outcomes. Countries should develop several scenarios with and without NPs/PAs to inform policy.


Cold-Water Immersion for Hyperthermic Humans Wearing American Football Uniforms.
Current treatment recommendations for American football players with exertional heatstroke are to remove clothing and equipment and immerse the body in cold water. It is unknown if wearing a full American football uniform during cold-water immersion (CWI) impairs rectal temperature (Trec) cooling or exacerbates hypothermic afterdrop. To determine the time to cool Trec from 39.5°C to 38.0°C while participants wore a full American football uniform or control uniform during CWI and to determine the uniform's effect on Trec recovery postimmersion. Crossover study. Laboratory. A total of 18 hydrated, physically active, unacclimated men (age = 22 ± 3 years, height = 178.8 ± 6.8 cm, mass = 82.3 ± 12.6 kg, body fat = 13% ± 4%, body surface area = 2.0 ± 0.2 m(2)). Participants wore the control uniform (undergarments, shorts, crew socks, tennis shoes) or full uniform (control plus T-shirt; tennis shoes; jersey; game pants; padding over knees, thighs, and tailbone; helmet; and shoulder pads). They exercised (temperature approximately 40°C, relative humidity approximately 35%) until Trec reached 39.5°C. They removed their T-shirts and shoes and were then immersed in water (approximately 10°C) while wearing each uniform configuration; time to cool Trec to 38.0°C (in minutes) was recorded. We measured Trec (°C) every 5 minutes for 30 minutes after immersion. Time to cool from 39.5°C to 38.0°C and Trec. The Trec cooled to 38.0°C in 6.19 ± 2.02 minutes in full uniform and 8.49 ± 4.78 minutes in control uniform (t17 = -2.1, P = .03; effect size = 0.48) corresponding to cooling rates of 0.28°C·min(-1) ± 0.12°C·min(-1) in full uniform and 0.23°C·min(-1) ± 0.11°C·min(-1) in control uniform (t17 = 1.6, P = .07, effect size = 0.44). The Trec postimmersion recovery did not differ between conditions over time (F1,17 = 0.6, P = .59). We speculate that higher skin temperatures before CWI, less shivering, and greater conductive cooling explained the faster cooling in full uniform. Cooling rates were considered ideal when the full uniform was worn during CWI, and wearing the full uniform did not cause a greater postimmersion hypothermic afterdrop. Clinicians may immerse football athletes with hyperthermia wearing a full uniform without concern for negatively affecting body-core cooling.


Antiangiogenic treatment with thrombospondin-1 enhances primary tumor radiation response and prevents growth of dormant pulmonary micrometastases after curative radiation therapy in human melanoma xenografts.
Thrombospondin-1 (TSP-1) is a potent antiangiogenic factor that has been shown to inhibit tumor growth by preventing endothelial cells from responding to a wide variety of angiogenic stimulators. We have demonstrated previously that D-12 primary tumors (human melanoma xenografts) suppress the growth of their spontaneous pulmonary micrometastases by secreting TSP-1 into the blood circulation. The same tumor model was used in the present work to study antitumor effects of combined radiation therapy and antiangiogenic treatment with TSP-1. Curative radiation treatment of D-12 primary tumors resulted in rapid growth of previously dormant micrometastases. Growth of dormant micrometastases could be prevented by treating the host mice with exogenous TSP-1 after the radiation treatment. Treatment with exogenous TSP-1 after subcurative radiation treatment reduced the growth rate of recurrent primary tumors in addition to suppressing metastatic growth. TSP-1 suppressed tumor growth at both primary and metastatic sites by inducing apoptosis in tumor-associated microvascular endothelial cells. Treatment with exogenous TSP-1 before radiation treatment enhanced the antitumor effect of the radiation treatment. The radiopotentiation by TSP-1 involved at least two distinctly different mechanisms, i.e., TSP-1 reduced the fraction of radiobiologically hypoxic parenchymal tumor cells and increased the radiation sensitivity of the tumor microvasculature by promoting radiation-induced endothelial cell apoptosis. In conclusion, the present preclinical study showed that TSP-1 has antiangiogenic, antimetastatic, and radiopotentiating properties that merit additional investigation in clinical studies.


Skin blood flow reduction induced by cigarette smoking: role of vasopressin.
The effect of vasopressin released by cigarette smoking on blood pressure (BP), heart rate (HR), and skin blood flow (SBF) was investigated in 12 normotensive habitual smokers. At a 1-wk interval, each subject smoked within 10 min two cigarettes before and after intravenous injection of either the specific vascular vasopressin antagonist d(CH2)5Tyr(Me)AVP (5 micrograms/kg) or its vehicle administered in double-blind fashion. SBF was assessed with a laser Doppler flowmeter. Smoking increased plasma vasopressin (P less than 0.01). In six subjects subsequently treated with the antagonist, plasma vasopressin rose to greater than 10 pg/ml and SBF fell by 18.2 +/- 4.8%. This SBF reduction was prevented by the vasopressin antagonist. In contrast, the vehicle had no effect. In the 24 studies taken together, there was a significant correlation (r = -0.60, P less than 0.01) between the SBF decrease during the first smoking period and the plasma vasopressin levels measured afterwards. The BP and HR rise caused by smoking was not modified by the antagonist. Thus it appears that the decrease in SBF induced by smoking is due to enhanced vasopressin secretion.


Modeling the influence of resonance stabilization on the kinetics of hydrogen abstractions.
Resonance stabilization of the transition state is one of the key factors in modeling the kinetics of hydrogen abstraction reactions between hydrocarbons. A group additive model is developed which allows the prediction of rate coefficients for bimolecular hydrogen abstraction reactions over a broad range of hydrocarbons and hydrocarbon radicals between 300 and 1300 K. Group additive values for 50 groups are determined from rate coefficients determined using the high level CBS-QB3 ab initio method, corrected for tunneling and the hindered internal rotation around the transitional bond. Resonance and hyperconjugative stabilization of the transition state is accounted for by introducing 4 corrections based on the structure of the reactive moiety of the transition state. The corrections, fitted to a set of 28 reactions, are temperature-independent and reduce the mean absolute deviation on E(a) to 0.7 kJ mol(-1) and to 0.05 for log A. Tunneling contributions are accounted for by using a fourth order polynomial in the activation energy. Final validation for 19 reactions yields a mean factor of deviation between group additive prediction and ab initio calculation of 2.4 at 300 K and 1.8 at 1000 K. In comparison with 6 experimental rate coefficients (600-719 K), the mean factor of deviation is less than 3.


Examining Inhibitory Affective Processing Within the Rostral Anterior Cingulate Cortex Among Abstinent Cannabis-Using Adolescents and Young Adults.
Cannabis use has been associated with deficits in self-regulation, including inhibitory control. Cannabis users have previously exhibited both structural and functional deficits in the rostral anterior cingulate cortex (rACC), a region involved in self-regulation of emotional response and inhibitory control. The present study aimed to examine whether abstinent cannabis users demonstrated abnormal functional activation and connectivity of the bilateral rACC during an emotional inhibitory processing task, and whether gender moderated these relationships. Cannabis-using (N = 34) and non-using (N = 32) participants ages 16-25 underwent at least 2-weeks of monitored substance use abstinence (excluding tobacco) and fMRI scanning while completing a Go/No-go task using fearful and calm emotional faces as non-targets. Multiple linear regression and ANCOVA were used to determine if cannabis group status was related to rACC activation and context-dependent functional connectivity, and whether gender moderated these relationships. Results showed decreased bilateral rACC activation in cannabis users during fearful response inhibition, although groups did not show any context-dependent connectivity differences between the left or right rACC during calm or fearful inhibition. Gender findings revealed that cannabis-using females compared to males did show aberrant connectivity between the right rACC and right cerebellum. These results are consistent with literature demonstrating aberrant structural and functional rACC findings and suggest that chronic cannabis use may disrupt typical rACC development-even after abstinence-potentially conferring risk for later development of mood disorders. Marginal gender-specific connectivity findings bolster continued findings regarding female vulnerability to effects of cannabis on cognition and affect. Findings should be assessed in longitudinal studies to determine causality and timing effects.


S-Allyl cysteine alleviates inflammation by modulating the expression of NF-κB during chromium (VI)-induced hepatotoxicity in rats.
Hexavalent chromium (Cr (VI)) is a common environmental pollutant. Cr (VI) exposure can lead to severe damage to the liver, but the preventive measures to diminish Cr (VI)-induced hepatotoxicity need further study. S-allyl cysteine (SAC) is a constituent of garlic ( Allium sativum) and has many beneficial effects to humans and rodents. In this study, we intended to analyze the mechanistic role of SAC during Cr (VI)-induced hepatotoxicity. Male Wistar albino rats were induced with 17 mg/kg body weight to damage the liver. The Cr (VI)-induced rats were treated with 100 mg/kg body weight of SAC as an optimum dosage to treat hepatotoxicity. We observed that the levels of oxidants, lipid peroxidation and hydroxyl radical (OH•) were increased, and enzymatic antioxidants such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase were found to be decreased in Cr (VI)-induced rats. While treated with SAC, the levels of oxidants were decreased and enzymatic antioxidants were significantly ( p < 0.05) increased. Lysosomal enzyme activities were increased in Cr (VI)-induced rats and on treatment with SAC, the activities were significantly decreased. The expressions of nuclear factor-kappa B (p65-NF-κB), tumor necrosis factor α (TNF-α), and inducible nitric oxide synthase (iNOS) were increased during induction with Cr (VI). Subsequent administration of SAC to animals showed a decrease in the expressions of NF-κB, TNF-α, and iNOS. Results obtained from this study clearly demonstrated that SAC protects the liver cells from the Cr (VI)-induced free radical damage.


Knee Cell-Based Cartilage Restoration.
As our patients become more physically active at all ages, the incidence of injuries to articular cartilage is increasing causing significant pain and disability. The intrinsic healing response of articular cartilage is poor because of its limited vascular supply and capacity for chondrocyte division. Nonsurgical management for the focal cartilage lesion is successful in the majority of patients. Those patients who fail conservative management may be candidates for a cartilage reparative or reconstructive procedure. The type of treatment available depends on a multitude of lesion-specific and patient-specific variables. First-line therapies for isolated cartilage lesions have demonstrated good clinical results in the correct patient, but typically repair cartilage with fibrocartilage, which has inferior stiffness, inferior resilience, and poorer wear characteristics. Advances in cell-based cartilage restoration have provided the surgeon a means to address focal cartilage lesions utilizing mesenchymal stem cells, chondrocytes, and biomimetic scaffolds to restore hyaline cartilage.


[Surgical management of the congenital atresia of the ear].
This study presents a 15 year experience of surgical treatment in 47 patients with congenital atresia of the ear. Emphasizing on the importance of pre-surgical evaluation of the patients for a better prognostic indicator. The ages of 40 patients were between 5-15 years. 24 patients (61%) had complete atresia and 23 (49%) incomplete. 27 patients (57%) with bilateral disease and 20 (43%) unilateral. Auditory test before surgery showed a conductive hearing loss of approximately 60 decibels (dB) and one case with total sensorineural hearing loss. There was an auditory average gain of 28.3 dB. 3 patients (6.4%) had associated cholesteatoma. Complications presented: Closure of the external auditory canal in 6 patients (12.7%), and facial nerve paralysis in 2 patients with recovery. The different surgical technics are described. In conclusion, for this type of surgery, combination between the otologist and plastic surgeon should be done for better results.


Effect of formulation and baking conditions on the structure and development of non-enzymatic browning in biscuit models using images.
The aim of this research was to determine the effect of composition (dietary fiber = DF, fat = F, and gluten = G) and baking time on the target microstructural parameters that were observed using images of potato and wheat starch biscuits. Microstructures were studied Scanning Electron Microscope (SEM). Non-enzymatic browning (NEB) was assessed using color image analysis. Texture and moisture analysis was performed to have a better understanding of the baking process. Analysis of images revealed that the starch granules retained their native form at the end of baking, suggesting their in complete gelatinization. Granules size was similar at several different baking times, with an average equivalent diameter of 9 and 27 µm for wheat and potato starch, respectively. However, samples with different levels of DF and G increased circularity during baking to more than 30%, and also increasing hardness. NEB developed during baking, with the maximum increase observed between 13 and 19 min. This was reflected in decreased luminosity (L*) values due to a decrease in moisture levels. After 19 min, luminosity did not vary significantly. The ingredients that are used, as well as their quantities, can affect sample L* values. Therefore, choosing the correct ingredients and quantities can lead to different microstructures in the biscuits, with varying amounts of NEB products.


The chloroplast small heat-shock protein oligomer is not phosphorylated and does not dissociate during heat stress in vivo.
Plants synthesize several classes of small (15- to 30-kD monomer) heat-shock proteins (sHSPs) in response to heat stress, including a nuclear-encoded, chloroplast-localized sHSP (HSP21). Cytosolic sHSPs exist as large oligomers (approximately 200-800 kD) composed solely or primarily of sHSPs. Phosphorylation of mammalian sHSPs causes oligomer dissociation, which appears to be important for regulation of sHSP function. We examined the native structure and phosphorylation of chloroplast HSP21 to understand this protein's basic properties and to compare it with cytosolic sHSPs. The apparent size of native HSP21 complexes was > 200 kD and they did not dissociate during heat stress. We found no evidence that HSP21 or the plant cytosolic sHSPs are phosphorylated in vivo. A partial HSP21 complex purified from heat-stressed pea (Pisum sativum L.) leaves contained no proteins other than HSP21. Mature recombinant pea and Arabidopsis thaliana HSP21 were expressed in Escherichia coli, and purified recombinant Arabidopsis HSP21 assembled into homo-oligomeric complexes with the same apparent molecular mass as HSP21 complexes observed in heat-stressed leaf tissue. We propose that the native, functional form of chloroplast HSP21 is a large, oligomeric complex containing nine or more HSP21 subunits, and that plant sHSPs are not regulated by phosphorylation-induced dissociation.


Fourth International Workshop on Haploidentical Transplants, Naples, Italy, July 8-10, 2004.
Many patients with high-risk hematological malignancies or with incurable inborn errors do not have an HLA-matched sibling and cannot find an HLA-matched donor for an allogeneic hematopoietic stem cell transplantation (SCT). Transplantation strategies using mismatched haploidentical family donors have been an important development. Although the procedure has saved patients from certain death, it is still beset by major problems like life-threatening infections--due to profound immunodeficiency following T-cell depletion and to disease relapse. At every International Workshop on Haploidentical Transplants, new data are presented, showing how scientists are attempting to improve the outcomes of mismatched transplants while reducing the severity of complications.The fourth Workshop continues in this tradition of presenting ground-breaking research. It opened with presentations of the current results with unrelated volunteer and umbilical cord blood transplants and proceeded to a session with the results of haploidentical transplants in the world with series of patients with high-risk acute leukemia, ranging in number from well over 100 in Perugia, Italy, and 80 in Haifa, Israel, to smaller groups in Europe and the United States. The session on graft engineering presented the latest results in the search for the optimal graft. The graft-vs.-leukemia effect in the haploidentical transplant was discussed in depth. Subsequently, attention focussed on one of the major problems in haploidentical transplant, that is, the delay in immunological recovery. The Workshop closed with presentations on tolerance induction that was followed by the results of ongoing registration studies being performed by the Italian GIMEMA group and the European Bone Marrow Transplant group.


Polysaccharide-Based Bioink Formulation for 3D Bioprinting of an In Vitro Model of the Human Dermis.
Limitations in wound management have prompted scientists to introduce bioprinting techniques for creating constructs that can address clinical problems. The bioprinting approach is renowned for its ability to spatially control the three-dimensional (3D) placement of cells, molecules, and biomaterials. These features provide new possibilities to enhance homology to native skin and improve functional outcomes. However, for the clinical value, the development of hydrogel bioink with refined printability and bioactive properties is needed. In this study, we combined the outstanding viscoelastic behavior of nanofibrillated cellulose (NFC) with the fast cross-linking ability of alginate (ALG), carboxymethyl cellulose (CMC), and encapsulated human-derived skin fibroblasts (hSF) to create a bioink for the 3D bioprinting of a dermis layer. The shear thinning behavior of hSF-laden bioink enables construction of 3D scaffolds with high cell density and homogeneous cell distribution. The obtained results demonstrated that hSF-laden bioink supports cellular activity of hSF (up to 29 days) while offering proper printability in a biologically relevant 3D environment, making it a promising tool for skin tissue engineering and drug testing applications.


Synopsis of preterm birth genetic association studies: the preterm birth genetics knowledge base (PTBGene).
Our goal wasto produce a field synopsis of genetic associations with preterm birth and to set up a publicly available online database summarizing the data. We performed a systematic review and meta-analyses to identify genetic associations with preterm birth. We have set up a publicly available online database of genetic association data on preterm birth called PTBGene (http://ric.einstein.yu.edu/ptbgene/index.html) and report on a structured synopsis thereof as of December 1, 2008. Data on 189 polymorphisms in 84 genes have been included and 36 meta-analyses have been performed. Five gene variants (4 in maternal DNA, one in newborn DNA) have shown nominally significant associations, but all have weak epidemiological credibility. After publishing this field synopsis, the PTBGene database will be regularly updated to keep track of the evolving evidence base of genetic factors in preterm birth with the goal of promoting knowledge sharing and multicenter collaboration among preterm birth research groups.


Adverse drug reactions among children over a 10-year period.
This study was designed to evaluate (1) the incidence and common types of adverse drug reactions among hospitalized children, (2) the frequency of adverse drug reaction reporting by health care providers, and (3) the follow-up processes resulting from adverse drug reactions. A retrospective cohort study of pediatric patients who experienced an adverse drug reaction between January 1, 1995, and December 31, 2004, was conducted at a community-based, tertiary care, children's teaching hospital. A total of 1087 adverse drug reactions were reported; the overall incidence was 1.6%. The severity of most adverse drug reactions was low (levels 1-3: 89%; high levels 4-6: 11%). Adverse drug reactions with low severity were significantly more common in both the general pediatric unit and the NICU. Adverse reactions resulting from use of antibiotics (particularly penicillins, cephalosporins, and vancomycin) were usually mild. In contrast, adverse drug reactions rated high in severity were significantly more common among reactions that led to hospital admission or occurred during surgery and among certain drug classes, including anticonvulsants and antineoplastic agents. Adverse drug reactions were reported by pharmacists (89%), nurses (10%), and physicians (< 1%). Although documentation of physician notification occurred for 93% of adverse drug reactions, only 29% of cases were documented in the patient's medical chart, 13% included follow-up education for individuals involved, and 10% were updated in the allergy profile of the hospital computer system. Measures to improve detection and reporting of adverse drug reactions by all health care professionals should be undertaken, to enhance our understanding of the nature and impact of these reactions in children.


Evolution of the amygdaloid complex in vertebrates, with special reference to the anamnio-amniotic transition.
Numerous studies over the last few years have demonstrated that the amygdaloid complex in amniotes shares basic developmental, hodological and neurochemical features. Furthermore, homologous territories of all the main amygdaloid subdivisions have been recognized among amniotes, primarily highlighted by the common expression patterns for numerous developmental genes. Thus, derivatives from the lateral pallium, ventral pallium and subpallium constitute the fundamental parts of the amygdaloid complex. With the development of new technical approaches, study of the precise neuroanatomy of the telencephalon of the anuran amphibians (anamniotes) has been possible. Current embryological, hodological and immunohistochemical evidence strongly suggests that most of the structures present in amniotes are recognizable in these anamniotes. These investigations have yielded enough results to support the notion that the organization of the anuran amygdaloid complex includes subdivisions with their origin in ventral pallial and subpallial territories; a strong relationship with the vomeronasal and olfactory systems; abundant intra-amygdaloid connections; a main output centre involved in the autonomic system; recognizable amygdaloid fibre systems; and distinct chemoarchitecture. Therefore, the new ideas regarding the amygdaloid evolution based on the recent findings in anamniotes, and especially in anurans, strongly support the notion that basic amygdaloid structures were present at least in the brain of ancestral tetrapods organized following a basic plan shared by tetrapods.


Double peak sensory responses: effects of capsaicin.
The aim of this study is to verify whether degeneration of skin receptors or intradermal nerve endings by topical application of capsaicin modifies the double peak response obtained by submaximal anodal stimulation. Five healthy volunteers topically applied capsaicin to the finger-tip of digit III (on the distal phalanx) four times daily for 4-5 weeks. Before and after local capsaicin applications, we studied the following electrophysiological findings: compound sensory action potential (CSAP), double peak response, sensory threshold and double peak stimulus intensity. Local capsaicin application causes disappearance or decrease of the second component of the double peak, which gradually increases after the suspension of capsaicin. Conversely, no significant differences were observed for CSAP, sensory threshold and double peak stimulus intensity. This study suggests that the second component of the double peak may be a diagnostic tool suitable to show an impairment of the extreme segments of sensory nerve fibres in distal sensory axonopathy in the early stages of damage, when receptors or skin nerve endings are impaired but undetectable by standard nerve conduction studies.


Effect of various veterinary procedures on plasma concentrations of cortisol, luteinising hormone and prostaglandin F2 alpha metabolite in the cow.
Five commonly practised veterinary procedures were studied: Palpation per rectum of the reproductive tract, intramuscular injection, single venepuncture, repeated venepuncture and jugular vein catheterisation. Plasma cortisol concentrations increased from baseline values of approximately 2 ng/ml to maximum mean values between 6.5 +/- 2.5 ng/ml and 13.8 +/- 5.6 ng/ml approximately 13 to 27 minutes after each manipulation. Baseline values occurred approximately 80 minutes later. In the control bleeding periods unacclimatised cows initially had high values of plasma cortisol (5 to 10 ng/ml) which returned to baseline after two hours, ie, before beginning any procedure. There were no statistically significant changes in luteinising hormone concentrations. The concentration of 13,14 dihydro, 15-keto prostaglandin F2 alpha (PGFM) increased from 61.0 +/- 4.6 pg/ml to 209.8 +/- 152.1 pg/ml in three out of five cows palpated on days 16 and 19 of the oestrous cycle. Increases did not occur in five other cows palpated during the follicular phase, nor in five cows palpated on day 12. However, after palpation on day 8, one animal did have concentrations of PGFM similar to those occurring during spontaneous release on days 18 to 20 of the oestrous cycle.


Maternal perception of weight status in first-born Australian toddlers aged 12-16 months--the NOURISH and SAIDI cohorts.
The preference amongst parents for heavier infants is in contrast to obesity prevention efforts worldwide. Parents are poor at identifying overweight in older children, but few studies have investigated maternal perception of weight status amongst toddlers and none in the Australian setting. Mothers (n = 290) completed a self-administered questionnaire at child age 12-16 months, defining their child's weight status as underweight, normal weight, somewhat overweight or very overweight. Weight-for-length z-score was derived from measured weight and length, and children categorized as underweight, normal weight, at risk overweight or obese (WHO standards). Objective classification was compared with maternal perception of weight status. Mean weight-for-length z-score was compared across categories of maternal perception using one-way ANOVA. Multinomial logistic regression was used to determine child or maternal characteristics associated with inaccurate weight perception. Most children (83%) were perceived as normal weight. Twenty nine were described as underweight, although none were. Sixty-six children were at risk of overweight, but 57 of these perceived as normal weight. Of the 14 children who were overweight, only 4 were identified as somewhat overweight by their mother. Compared with mothers who could accurately classify their normal weight child, mothers who were older had higher odds of perceiving their normal weight child as underweight, while mothers with higher body mass index had slightly higher odds of describing their overweight/at risk child as normal weight. The leaner but healthy weight toddler was perceived as underweight, while only the heaviest children were recognized as overweight. Mothers unable to accurately identify children at risk are unlikely to act to prevent further excess weight gain. Practitioners can lead a shift in attitudes towards weight in infants and young children, promoting routine growth monitoring and adequate but not rapid weight gain.


[Prezygotic selection of male gametes in laboratory mice].
Frequency of aneuploid male gametes was related to the number of early embryos with numeral chromosomal aberrations in progeny of male mice, heterozygous for the Robertsonian translocations T1 (15.6) AID, and T1 (17.8) IEM. In mice heterozygous for translocation T1 (15.6) AID, the number of spermatocytes II with NF=21 was very low and corresponded to the number of trisomic embryos at stages of major organogenesis. In mice heterozygous for T1 (17.8) IEM translocation, the number of aneuploid gametes was 26.2%, but neither embryo of their progeny, on the 8 12th days of gestation, was found to be trisomic. The involvement of chromosomes 8 and 17 in the control of postmeiotic stages of spermatogenesis in mice is suggested.


Bcl-2 and Bcl-XL are indispensable for the late phase of mast cell development from mouse embryonic stem cells.
The aim of this study was to determine the importance of the prosurvival factors Bcl-2 and Bcl-XL for mast cell development and survival. bcl-x(-/-) and bcl-2(-/-) mouse embryonic stem cells were maintained in medium supplemented with either interleukin (IL)-3 or IL-3 in combination with stem cell factor (SCF) to favor mast cell development. The development of Bcl-2 family deficient embryonic stem cell-derived mast cells (ESMCs) was monitored and Bcl-2 family gene expression and cell numbers were analyzed. Deficiency in either bcl-x or bcl-2 totally inhibited the development of ESMCs when IL-3 alone was used as a mast cell growth factor. Intriguingly, when IL-3 was used in combination with SCF, the ESMCs developed normally the first 2 weeks but thereafter the cell numbers dropped drastically. The remaining ESMCs express mouse mast cell protease 1, suggesting a mucosal-like phenotype. ESMCs lacking bcl-x or bcl-2 exhibited strong expression of A1, another prosurvival Bcl-2 family member. For the first time we provide direct evidence that both bcl-x and bcl-2 are indispensable for mast cell survival during the late phase of their development.