Transcriptional units for ribosomal proteins in yeast.
The effects of ultraviolet irradiation on the rates of synthesis of individual ribosomal proteins in yeast were examined and compared with the ultraviolet sensitivities of the synthesis of other yeast proteins. It was found that the synthesis of yeast ribosomal proteins is much more sensitive to ultraviolet irradiation than that of other yeast cellular proteins. Taking into account the half-life of yeast mRNA, the results obtained indicate that the genes coding for ribosomal proteins form part of long transcriptional units, which are much longer than the DNA stretch needed to code for a ribosomal protein of average molecular weight. Saturation hybridization of total poly(A)-containing mRNA with yeast nuclear DNA revealed that as much as 30% of DNA is complementary to yeast mRNA. Thus, the primary transcript of a protein gene on the average is about 1.7 times the length of the actual messenger. On the basis of the various experimental data we suggest a clustering of the yeast ribosomal protein genes in a number of common transcriptional units.


Comparative ultrastructure analysis of radiation-induced radioresistant laryngeal cancer hep-2 cell line.
Radioresistance is one of the main reasons for the failure of radiotherapy in laryngeal cancer. However, the mechanisms of radioresistance of tumor cells have remained elusive. This study was conducted to identify the ultrastructural changes of radiation-induced radioresistant laryngeal cancer hep-2 cell line. First, a radioresistant hep-2R cell line was generated after prolonged exposure to γ-rays for 60 Gy (6 Gy/day, 2 days/week) and was confirmed by clonogenic assay. Next, the ultrastructural differences between hep-2R cells and hep-2 cells were compared by transmission electron microscopy. Finally, the results showed that hep-2R cells showed significant resistance to radiation compared with parental hep-2 cells. Increased cell nucleus atypia, more rough endoplasmic reticulum and less mitochondria were observed in hep-2R cells. The amount of microvilli of hep-2R was similar to hep-2 cell. In summary, these ultrastructural differences revealed the morphological mechanism that hep-2R cells had stronger radioresistance than hep-2 cells.


Clinical evaluation of high-molecular-weight sodium hyaluronate for the treatment of patients with periarthritis of the shoulder.
The clinical efficacy and tolerability of sodium hyaluronate, a high-molecular-weight hyaluronic acid preparation, in the treatment of patients with periarthritis of the shoulder was investigated in a multicenter, cooperative study. Of 70 patients seen at 12 participating centers between April and September 1992, follow-up was possible in 62 (28 men and 34 women; mean age, 65.4 years). Patients received sodium hyaluronate 25 mg (1% solution in an ampule) once weekly for 5 weeks by injection into the glenoid cavity or the subacromial bursa. If periarthritis resolved during this period, treatment was stopped; if periarthritis had not fully resolved by the end of the 5 weeks, weekly or biweekly treatment was continued as appropriate. The mean treatment period was 8.16 +/- 0.88 weeks, and the mean total administered dose was 6.05 +/- 0.61 ampules. Rates of improvement over pretreatment values were 75.0% for pain at rest, 73.7% for pain on motion, and 78.8% for pain on pressure. Significant increases in joint angles (measured as components of joint range of motion after administration of sodium hyaluronate) were observed; activities of daily living also improved significantly (P < 0.05 to P < 0.001). No adverse reactions were observed. These results strongly suggest that sodium hyaluronate is an effective agent for the treatment of patients with periarthritis of the shoulder.


Validation and noise robustness assessment of microscopic anisotropy estimation with clinically feasible double diffusion encoding MRI.
Double diffusion encoding (DDE) MRI enables the estimation of microscopic diffusion anisotropy, yielding valuable information on tissue microstructure. A recent study proposed that the acquisition of rotationally invariant DDE metrics, typically obtained using a spherical "5-design," could be greatly simplified by assuming Gaussian diffusion, facilitating reduced acquisition times that are more compatible with clinical settings. Here, we aim to validate the new minimal acquisition scheme against the standard DDE 5-design, and to quantify the proposed method's noise robustness to facilitate future clinical use. DDE MRI experiments were performed on both ex vivo and in vivo rat brains at 9.4 T using the 5-design and the proposed minimal design and taking into account the difference in the number of acquisitions. The ensuing microscopic fractional anisotropy (μFA) maps were compared over a range of b-values up to 5000 s/mm2 . Noise robustness was studied using analytical calculations and numerical simulations. The minimal protocol quantified μFA at an accuracy comparable to the estimates obtained by means of the more theoretically robust DDE 5-design. μFA's sensitivity to noise was found to strongly depend on compartment anisotropy and tensor magnitude in a nonlinear manner. When μFA < 0.75 or when mean diffusivity is particularly low, very high signal-to-noise ratio is required for precise quantification of µFA. Our work supports using DDE for quantifying microscopic diffusion anisotropy in clinical settings but raises hitherto overlooked precision issues when measuring μFA with DDE and typical clinical signal-to-noise ratio.


Theorems and implications of a model-independent elimination/distribution function decomposition of linear and some nonlinear drug dispositions. II. Clearance concepts applied to the evaluation of distribution kinetics.
The disposition decomposition approach is employed to derive clearance parameters descriptive of drug distribution kinetics. The name distribution clearance, CLd, is given to a characteristic constant of linear and some nonlinear pharmacokinetic systems. CLd is the clearance associated with the steady-state rate of drug transfer from the peripheral tissues to the systemic circulation. Also introduced is the elimination clearance, CLe, which is associated with the total drug transfer rate from the systemic circulation in linear systems. Estimates of CLd and CLe are presented for several drugs.


Emergency department attendance by children at risk of abuse.
Frequency of emergency department (ED) attendance has long been thought to be a risk factor for child abuse. The aim of this study was to test this assumption by comparing the ED attendances of at-risk children (before being placed on a child protection register) with the attendances of an age-matched control group (before an index attendance) Method: A group of 220 children (aged 0-12 years inclusive) were identified from the two child protection registers in the Shropshire area in 2006. The ED attendances of these children in the 2 years before registration were identified using the computer records of the two local EDs. A control group of 150 children for each year of age (0-12 years inclusive) was then identified from ED attendances between October and December 2006. The attendances of these children in the 2 years before this index attendance were obtained. The data for these two groups of children were then compared. The at-risk children did not attend the EDs more frequently than did the children in the control group. The identification of children who attend EDs frequently may be useful for other reasons but is unlikely to be an effective way to detect child abuse.


[Anxiolytic effects of lisuride and its agonistic action to central 5-HT1A receptors].
Effects of lisuride, a derivative of ergot alkaloid, on central 5-HT1A receptors were investigated biochemically, behaviorally and electroencephalographically (EEG) in rats and rabbits. Effects of lisuride in water-lick conflict tests were also investigated in rats. Lisuride was found to strongly inhibit the bindings of [3H]8-OH-DPAT to 5-HT1A receptors in the raphe nucleus, hippocampus, cortex, amygdala and hypothalamus of rat brain. Inhibitory effects of lisuride on bindings of [3H]8-OH-DPAT in the hippocampus was almost the same as that of 5-HT (Ki = 0.5 nM) and stronger than those of the 5-HT agonist 5-MeO-DMT (Ki = 2.1 nM) or other ergot derivatives (bromocriptine and pergolide, Ki = 3.0 nM). Lisuride (0.1-0.5 mg/kg, i.p.), like 8-OH-DPAT, dose-dependently induced a 5-HT behavioral syndrome in rats. Lisuride affected locomotor activity in rats, whereas 8-OH-DPAT did not. In hippocampal EEG of rabbits, lisuride (0.01-0.03 mg/kg, i.v.), like 8-OH-DPAT and diazepam, dose-dependently inhibited rhythmical slow activity (RSA) induced by acoustical stimulation (3100 Hz) and also inhibited the RSA increased by administration of anxiogenic FG7142. In water-lick conflict tests, lisuride (0.05-0.1 mg/kg, i.p.), like diazepam, increased the number of shocks. These findings indicated that lisuride acts as a strong agonist on central 5-HT1A receptors and suggested that lisuride might be a potential anxiolytic.


[Economical efficacy of physical exercises for osteoporosis].
From mean 12.5% increase of lumbar bone mineral density by one year's physical training, it is calculated to reduce the frequency of femoral neck fracture by 80% and the rate of falls by 50%. And 60% reduction of femoral neck fracture by physical exercise gains about 200 billion yen of medical and care costs annually, because a patient with femoral neck fracture consumes mean 1,320,000 yen for medical treatments and about 19% of the patients become bed-ridden state after the treatments for mean 5 years.


Mine rescuers' heat load during the expenditure of physical effort in a hot environment, using ventilated underwear and selected breathing apparatus.
Mine rescuers' heat load under the same physical effort load (25% of the maximal oxygen uptake), using three types of breathing apparatus, in newly developed heat-removing underwear and outerwear was assessed for typical work conditions of mine rescuers, under milder and harsher ambient conditions of 32 and 38 °C, respectively, both at relative humidity of 85% and air velocity of 1.0 m/s. Expending physical effort at the same load while using different kinds of breathing apparatus resulted in a similar heat load. Under both milder and harsher ambient conditions, heat storage and sweating intensity were greater than the average limit value recommended by hygienic standards, which indicates that the use of breathing apparatus significantly hinders heat exchange with the environment. The developed clothing for mine rescuers was highly rated, and was considered by most people to be better than that used currently.


Effects of flaxseed lignan and oil on bone health of breast-tumor-bearing mice treated with or without tamoxifen.
Previous studies showed that flaxseed lignan (secoisolariciresinol diglucoside, SDG) and oil (FO) inhibit established breast tumor growth in athymic mice with or without tamoxifen (TAM) treatment. TAM was found to increase bone mineral content (BMC) and density (BMD) in breast cancer patients. It is not known whether SDG or FO alone or combined with TAM affects bone health. Hence, the effects of SDG and FO, alone or in combination, on BMC, BMD, and biomechanical bone strength in ovariectomized athymic mice with established human breast tumors (MCF-7) treated with or without TAM were studied. In a factorial design, mice were divided into four non-TAM and four TAM groups. Each group consisted of mice fed a basal diet (BD), SDG (1 g/kg), FO (38.5 g/kg) or SDG + FO (combination) diets. The TAM group had TAM implants that provide a 5-mg TAM dose released over 60 d. TAM exerted an overall significant effect in increasing BMC, BMD, and biomechanical strength in femurs and lumbar vertebra. Without TAM treatment, SDG produced significant lower femur BMD (6%) while FO produced lower vertebrae BMC (8%) and BMD (6%). With TAM treatment, SDG and FO did not exert an effect on BMC and BMD at the femur or vertebra. SDG and FO produced no marked effect on biomechanical bone strength with or without TAM treatment. In conclusion, FS components did not significantly attenuate the positive effects on bone induced by TAM in this model system, indicating no apparent adverse effects on bone health.


Preoperative frailty is predictive of complications after major lower extremity amputation.
Preoperative clinical frailty is increasingly used as a surrogate for predicting postoperative outcomes. Patients undergoing major lower extremity amputation (LEA) carry a high risk of perioperative morbidity and mortality, including high 30-day mortality and readmission rates. We hypothesized that preoperative frailty would be associated with an increased risk of postoperative mortality and readmission. A retrospective review was performed for all patients who underwent transfemoral or transtibial amputation for any indication within a multi-institution system during a 5-year period. Standard demographics and all components of the Modified Frailty Index (mFI) were used to determine preoperative frailty status for each patient. The primary outcome was 30-day mortality, with secondary outcomes of 30-day readmission, unplanned revision, and composite adverse events. Among 379 patients who underwent LEA, the overall readmission and mortality rates for the group were 22.69% and 6.06%, respectively. Readmission rates increased with increasing mFI score: rates were 8.6%, 13.5%, 16.3%, 19.7%, 31.4%, and 37.0% for mFI scores of 0, 1, 2, 3, 4, and ≥5, respectively (P = .015). On multivariate logistic regression, only mFI (odds ratio, 1.49, 95% confidence interval, 1.24-1.77) and sex (odds ratio, 1.81, 95% confidence interval, 1.00-2.98) were significant predictors of 30-day readmission. Preoperative clinical frailty is associated with an increased 30-day readmission rate in patients undergoing LEA and should be incorporated into preoperative counseling and risk stratification, as well as postoperative planning and care.


The magnitude and time-dependence of the apoptotic response of normal and malignant cells subjected to ionizing radiation versus hyperthermia.
The purpose of the study was to examine the optimal time of exposure and dose of heat and ionizing radiation that results in the killing of human cancer cells in vitro via apoptosis vs. necrosis. Human mammary carcinoma, colorectal carcinoma and normal bovine capillary endothelial (BCE) cell lines were subjected to 20 Gy ionizing radiation and 6, 12, 24, and 72 h later assessed for apoptosis using detection of apoptotic bodies and caspase assays. Necrosis was detected by loss of cells from the surface and lactate dehydrogenase (LDH) release. The colorectal carcinoma cells were subjected to hyperthermia using temperatures ranging from 39 - 44 degrees C for 5, 15 or 45 min. exposures and at varying times post-treatment, apoptosis and necrosis were measured. In response to ionizing radiation, none of the cells underwent necrosis and some cell types apoptosed 24 and 72 h posttreatment. The colorectal cancer cells exhibited a steady increase of apoptosis at 6, 12, and 24 h. When these cells were exposed to 40 degrees C for 5 min, caspases increased within 6 h and a significant fraction (50%) of cells apoptosed. If the time of exposure to 40 degrees C was increased to 15 or 45 min, 80% and 100% of the dying cells apoptosed, respectively. A temperature of 39 degrees C did not cause cell death even after 45 min exposures. If heat was elevated to 42 or 44 degrees C, increased necrosis was observed with a corresponding decrease in apoptosis. These studies reveal time and temperature dependent in vitro cell responses to ionizing radiation and water-bath hyperthermia.


Interferon activation of latent natural killer cells and alteration in kinetics of target cell lysis: clinical implications for oral precancerous lesions.
Reduced natural killer cell activity was observed in patients with oral leukoplakia and submucous fibrosis compared with normal control subjects. However, the number of target binding lymphocytes was found to be normal in these precancers. Treatment of effector cells with interferon-alpha resulted in highly elevated active killer cell activity, although no change was observed in target binding lymphocyte counts. This finding could imply that precytotoxic cells that are activated by interferon exist in peripheral blood or that direct recruitment of a new cell population takes place. In addition, altered target lysis kinetics was observed, with interferon-activated killer cells demonstrating a tremendous lytic activation that is completed so quickly that a statistical kinetic analysis could not be accurately done. Because natural killer cell activity is an important effector system in immunosurveillance against tumors, its modulation with interferon may be an exciting clinical possibility in the control of malignant transformation or oral precancers.


Production of the Aspergillus aculeatus endo-1,4-beta-mannanase in A. niger.
The beta-mannanase gene (man1) from Aspergillus aculeatus MRC11624 (Izuka) was patented for application in the coffee industry. For production of the enzyme, the gene was originally cloned and expressed in Saccharomyces cerevisiae. However the level of production was found to be economically unfeasible. Here we report a 13-fold increase in enzyme production through the successful expression of beta-mannanase of Aspergillus aculeatus MRC11624 in Aspergillus niger under control of the A. niger glyceraldehyde-3-phosphate dehydrogenase promoter (gpd (P)) and the A. awamori glucoamylase terminator (glaA(T)). The effect of medium composition on mannanase production was evaluated, and it was found that the glucose concentration and the organic nitrogen source had an effect on both the volumetric enzyme activity and the specific enzyme activity. The highest mannanase activity levels of 16,596 nkat ml(-1) and 574 nkat mg(-1) dcw were obtained for A. niger D15[man1] when cultivated in a process-viable medium containing corn steep liquor as the organic nitrogen source and high glucose concentrations.


Treatment of unprotected left main stenoses with Palmaz-Schatz stenting.
Coronary artery bypass surgery remains the standard modality for revascularization of significant left main coronary artery stenoses. Despite major improvement in equipment and operator technique, balloon angioplasty of significant unprotected left main obstruction generally is contraindicated. When catheter intervention is required, Palmaz-Schatz stenting may offer better predictability of the acute result, and possibly improved long-term outcome.


Selective interactions of cationic porphyrins with G-quadruplex structures.
G-quadruplex DNA presents a potential target for the design and development of novel anticancer drugs. Because G-quadruplex DNA exhibits structural polymorphism, different G-quadruplex typologies may be associated with different cellular processes. Therefore, to achieve therapeutic selectivity using G-quadruplexes as targets for drug design, it will be necessary to differentiate between different types of G-quadruplexes using G-quadruplex-interactive agents. In this study, we compare the interactions of three cationic porphyrins, TMPyP2, TMPyP3, and TMPyP4, with parallel and antiparallel types of G-quadruplexes using gel mobility shift experiments and a helicase assay. Gel mobility shift experiments indicate that TMPyP3 specifically promotes the formation of parallel G-quadruplex structures. A G-quadruplex helicase unwinding assay reveals that the three porphyrins vary dramatically in their abilities to prevent the unwinding of both the parallel tetrameric G-quadruplex and the antiparallel hairpin dimer G-quadruplex DNA by yeast Sgs1 helicase (Sgs1p). For the parallel G-quadruplex, TMPyP3 has the strongest inhibitory effect on Sgs1p, followed by TMPyP4, but the reverse is true for the antiparallel G-quadruplex. TMPyP2 does not appear to have any effect on the helicase-catalyzed unwinding of either type of G-quadruplex. Photocleavage experiments were carried out to investigate the binding modes of all three porphyrins with parallel G-quadruplexes. The results reveal that TMPyP3 and TMPyP4 appear to bind to parallel G-quadruplex structures through external stacking at the ends rather than through intercalation between the G-tetrads. Since intercalation between G-tetrads has been previously proposed as an alternative binding mode for TMPyP4 to G-quadruplexes, this mode of binding, versus that determined by a photocleavage assay described here (external stacking), was subjected to molecular dynamics calculations to identify the relative stabilities of the complexes and the factors that contribute to these differences. The DeltaG(o) for the external binding mode was found to be driven by DeltaH(o) with a small unfavorable TDeltaS(o) term. The DeltaG(o) for the intercalation binding model was driven by a large TDeltaS(o) term and complemented by a small DeltaH(o) term. One of the main stabilizing components of the external binding model is the energy of solvation, which favors the external model over the intercalation model by -67.94 kcal/mol. Finally, we propose that intercalative binding, although less favored than external binding, may occur, but because of the nature of the intercalative binding, it is invisible to the photocleavage assay. This study provides the first experimental insight into how selectivity might be achieved for different G-quadruplexes by using structural variants within a single group of G-quadruplex-interactive drugs.


Baclofen and NOS inhibitors interact to evoke synergistic hypothermia in rats.
Our laboratory recently demonstrated that a drug combination of baclofen and L-NAME, a nonspecific nitric oxide synthase (NOS) inhibitor, evokes synergistic hypothermia in rats. These data are the first demonstration of synergy between a GABA agonist and NOS inhibitor. While the hypothermic synergy suggests a role for NOS in baclofen pharmacology, it is unclear whether the super-additive hypothermia is specific for baclofen and L-NAME or extends to drug combinations of baclofen and other NOS inhibitors. The site of action (central or peripheral) and isoforms of NOS that mediate the synergy are also unknown. Here, we confirm the hypothermic synergy with additional data and discuss potential mechanisms of the drug interaction. Baclofen (2.5, 3.5, 5 and 7.5 mg/kg, i.p.) was administered to rats by itself or with 7-nitroindazole (7-NI), a neuronal NOS inhibitor. 7-NI (10 mg/kg, i.p.) did not affect body temperature. For combined administration, 7-NI (10 mg/kg, i.p.) increased the relative potency of baclofen (F=18.9, P<0.05). The present data validate the hypothermic synergy caused by the drug combination of baclofen and L-NAME and implicate nNOS in the synergy. In a context broader than thermoregulation, NO production and transmission may play an important role in baclofen pharmacology.


A microwave spectroscopic and quantum chemical study of 3-butyne-1-selenol (HSeCH2CH2C[triple bond]CH).
The microwave spectrum of 3-butyne-1-selenol has been studied by means of Stark-modulation microwave spectroscopy and quantum chemical calculations employing the B3LYP/aug-cc-pVTZ and MP2/6-311++G(3df,3pd) methods. Rotational transitions attributable to the H80SeCH2CH2C[triple bond]CH and H78SeCH2CH2C[triple bond]CH isotopologues of two conformers of this molecule were assigned. One of these conformers possesses an antiperiplanar arrangement for the atoms Se-C-C-C, while the other is synclinal and seems to be stabilized by the formation of a weak intramolecular hydrogen bond between the hydrogen atom of the selenol group and the pi electrons of the CC triple bond. The energy difference between these conformers was determined to be 0.2(5) kJ/mol by relative intensity measurements, and the hydrogen-bonded form was slightly lower in energy.


[Cyclic nucleotide metabolic disorder in brain tumors].
The content of cAMP and cGMP as well as their ratio in the tissues of different cerebral tumors in humans were studied. The concentration of both cyclic nucleotides in tumors of the brain was found to vary widely. The richest information on disorders of metabolism of these substances may be gained from the ratio of cAMP and cGMP concentrations, which statistically is significantly reduced in the neoplasm as compared to the ratio in the cerebral tissue. Bearing in mind the principal possibility of correcting this ratio, further study of the metabolism of cyclic nucleotides in cerebral tumors holds promise for their treatment.


Closely related receptor complexes differ in their ABA selectivity and sensitivity.
The recent discovery of a variety of receptors has led to new models for hormone perception in plants. In the case of the hormone abscisic acid (ABA), which regulates plant responses to abiotic stress, perception seems to occur both at the plasma membrane and in the cytosol. The cytosolic receptors for ABA have recently been identified as complexes between protein phosphatases 2C (PP2C) and regulatory components (RCAR/PYR/PYL) that bind ABA. Binding of ABA to the receptor complexes inactivates the PP2Cs, thereby activating the large variety of physiological processes regulated by ABA. The Arabidopsis genome encodes 13 homologues of RCAR1 and approximately 80 PP2Cs, of which six in clade A have been identified as negative regulators of ABA responses. In this study we characterize a novel member of the RCAR family, RCAR3. RCAR3 was identified in a screen for interactors of the PP2Cs ABI1 and ABI2, which are key regulators of ABA responses. RCAR3 was shown to repress ABI1 and ABI2 in vitro, and to stimulate ABA signalling in protoplast cells. RCAR3 conferred greater ABA sensitivity to the PP2C regulation than RCAR1, whereas stereo-selectivity for (S)-ABA was less stringent with RCAR3 as compared with RCAR1. In addition, regulation of the protein phosphatase activity by RCAR1 and RCAR3 was more sensitive to ABA for ABI1 than for ABI2. Based on the differences we have observed in transcriptional regulation and biochemical properties, we propose a model whereby differential expression of the co-receptors and combinatorial assembly of the receptor complexes act in concert to modulate and fine-tune ABA responses.


[Mixed cryoglobulinemia].
The authors describe two clinical cases of cryoglobulinemia. A 70 years old woman, having skin ulcers on lower limbs, arthralgias, paresthesias and constitutional symptoms, for about 10 months. Exams revealed mild anemia, elevation of the biological parameters of inflammation and aminotransferases, positive cryoglobulin and rheumatoid factor in serum, and a severe reduction in C4 complement fraction. Hepatitis C virus (HCV) serology was negative. Idiopathic mixed cryoglobulinemia was diagnosed and corticosteroid therapy started. Given the lack of response, cyclophosphamide and plasmapheresis were added. Two weeks later the patient died in septic shock. The second case refers to a 41 years old female, with untreated hepatitis C who developed over a 6 month period petechiae and livedoid lesions on the lower limbs, peripheral neuropathy, and constitutional symptoms and was admitted with intestinal necrosis. Exams were consistent with the diagnosis of mixed cryoglobulinemia associated, with HCV. She started therapy with ribavirin and pegylated interferon-alpha, with improvement.


Direct and indirect effects of Growth Hormone Deficiency (GHD) on lung function in children: A mediation analysis.
Studies on pulmonary function tests (PFTs) in Growth Hormone Deficiency (GHD) children are lacking. The aims of this study were: (i) to investigate PFTs in GHD pre-pubertal children with respect to Controls, before starting Growth Hormone Therapy (GHT) (T0); (ii) to evaluate changes of PFTs in GHD vs Controls, after 1-year GHT (T1). For both aims the mediation analysis (MA) was applied to evaluate the extent to which the relationship between GHD and PFTs could be ascribed to a height-mediated (indirect) or a GH direct effect. 47 pre-pubertal GHD children (aged 5-14 years) underwent PFTs at T0 and T1. At T0, 47 healthy children matched for age and sex were enrolled as Controls. A MA was performed to assess the relationship between GHD and PFTs and height. Statistical analyses were performed using the statistical software R (https://cran.r-project.org/mirrors.html). A p-value <0.05 was considered significant. At T0, PFTs indices were significantly lower in GHD than in Controls. From T0 to T1 a significant improvement was found in PFTs. The percentages of the mediated effect on FVC, FEV1, FEF25-75% and TLC were <50% at T0, suggesting that the direct effect was prevalent. At T1, the percentages of the mediated effect for spirometry indices were ≥50%, indicating that the indirect (height-mediated) effect was the most relevant. The study shows that pre-pubertal children with GHD have an impairment of lung function not exclusively attributable to the indirect (height-mediated) effect, but also to the direct GH action which is mitigated after 1-year of GHT.


Intraosseous dentinogenic ghost cell tumor: a clinical report and literature update.
The dentinogenic ghost cell tumor (DGCT) is a relatively uncommon locally invasive neoplasm. This report describes the case of a 20-year-old Caucasian female patient who was diagnosed with an intraosseous DGCT in the left molar region of the mandible. Radiographic analysis revealed a well-circumscribed radiolucent lesion with focal radiopacity. Segmental resection of the mandible was performed by means of piezoelectric surgery. The histopathological examination yielded islands of odontogenic epithelium with clusters of ghost cells and dysplastic dentin, thus enabling the identification of the lesion as DGCT. Long-term surveillance of patients with intraosseous DGCTs is mandatory in order to detect in time any signs of recurrence. A literature update concerning intraosseous DGCTs is also provided.


Measurement of aldosterone in blood.
This chapter describes the measurement of aldosterone concentration in plasma or serum. Aldosterone specificity is improved by first extracting the aldosterone into dichloromethane. A radioimmunoassay is then performed on the dried and reconstituted extract using an anti-aldosterone antibody and iodinated aldosterone. The antibody-bound fraction is separated from the unbound fraction using activated charcoal. The notes section provides extra information for steps in the assay that can be problematic.


Monoamine receptor agonists, acting preferentially at presynaptic autoreceptors and heteroreceptors, downregulate the cell fate adaptor FADD in rat brain cortex.
FADD is a crucial adaptor of death receptors that can engage apoptosis or survival actions (e.g. neuroplasticity) through its phosphorylated form (p-FADD). Although FADD was shown to participate in receptor mechanisms related to drugs of abuse, little is known on its role in the signaling of classic neurotransmitters (dopamine, noradrenaline, and serotonin) in brain. This study assessed the modulation of FADD (and p-FADD/FADD ratio, as an index of neuroplasticity) and FLIP-L (a neuroprotective FADD interacting partner), as well as the role of MEK-ERK signaling, after activation of monoamine auto/heteroreceptors by selective agonists in rat cortex. Acute depletion of monoamines with reserpine, but not with AMPT or PCPA, reduced FADD (28%) and increased p-FADD/FADD ratio (1.34-fold). Activation of presynaptic α2A-adrenoceptors (UK-14304 and clonidine), 5-HT1A receptors (8-OH-DPAT), and D2 dopamine receptor (bromocriptine) dose-dependently decreased FADD (up to 54%) and increased p-FADD (up to 29%) and p-FADD/FADD ratios (up to 2.93-fold), through specific receptor mechanisms. Activation of rat 5-HT1B autoreceptor in axon terminals by CP-94253 did not modulate FADD forms. Activation of postsynaptic D1 dopamine receptor by SKF-81297 also reduced FADD (25%) and increased p-FADD (32%). Disruption of MEK-ERK activation with SL327 did not modify clonidine (α2A-adrenoceptor)-induced FADD inhibition, indicating that agonist effect was not dependent on ERK signaling. The various monoamine receptor agonists and antagonists did not alter FLIP-L content, or the activation of executioner caspase-3 and PARP-1 cleavage, indicating that the agonists attenuated apoptotic signals and promoted neuroplasticity through FADD regulation. These novel results indicate that inhibition of pro-apoptotic FADD adaptor could function as a common signaling step in the initial activation of monoamine receptors in the brain.


Osteoclast-derived coupling factors in bone remodeling.
In the bone remodeling process that takes place throughout the skeleton at bone multicellular units, intercellular communication processes are crucial. The osteoblast lineage has long been known to program osteoclast formation and hence resorption, but the preservation of bone mass and integrity requires tight control of remodeling. This needs local controls that ensure availability of mesenchymal precursors and the provision of local signals that promote differentiation through the osteoblast lineage. Some signals can come from growth factors released from resorbed bone matrix, and there is increasing evidence that the osteoclast lineage itself produces factors that can either enhance or inhibit osteoblast differentiation and hence bone formation. A number of such factors have been identified from predominantly in vitro experiments. The coupling of bone formation to resorption is increasingly recognized as a complex, dynamic process that results from the input of many local factors of cell and matrix origin that can either promote or inhibit bone formation.


Phase II evaluation of 1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitrosourea (PCNU, NSC 95466) in patients with metastatic colorectal cancer.
Thirty-two patients with measurable metastatic colorectal cancer refractory to 5-fluorouracil-containing regimens received PCNU (1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitrosourea, NSC 95466) on a single-day I.V. schedule administered every 6 weeks. Good-risk patients received PCNU at the starting dose of 100 mg/m2, while patients who had received radiotherapy or myelosuppressive drugs such as mitomycin C received an initial dose of 75 mg/m2. There were two partial remissions. Fifteen patients including four with minor tumor regression had disease stabilization. The dose-limiting toxicity was myelosuppression, with thrombocytopenia being more severe than neutropenia. The myelosuppression occurred late in the treatment cycle (days 25-32), was more severe with repeated treatments, and was more severe in patients who had a poor bone marrow reserve. Other toxicities were mild and infrequent. PCNU administered by the single-dose I.V. schedule as used in this study has very modest antitumor activity against colorectal cancer in humans.


Kinetic studies of water uptake and loss in glass-ionomer cements.
The water sorption and desorption behaviour of three commercial glass-ionomer cements used in clinical dentistry have been studied in detail. Cured specimens of each material were found to show slight but variable water uptake in high humidity conditions, but steady loss in desiccating ones. This water loss was found to follow Fick's law for the first 4-5 h. Diffusion coefficients at 22 degrees C were: Chemflex 1.34 x 10(-6) cm(2) s(-1), Fuji IX 5.87 x 10(-7) cm(2) s(-1), Aquacem 3.08 x 10(-6) cm(2) s(-1). At 7 degrees C they were: Chemflex 8.90 x 10(-7) cm(2) s(-1), Fuji IX 5.04 x 10(-7) cm(2) s(-1), Aquacem 2.88 x 10(-6) cm(2) s(-1). Activation energies for water loss were determined from the Arrhenius equation and were found to be Chemflex 161.8 J mol(-1), Fuji IX 101.3 J mol(-1), Aquacem 47.1 J mol(-1). Such low values show that water transport requires less energy in these cements than in resin-modified glass-ionomers. Fick's law plots were found not to pass through the origin. This implies that, in each case, there is a small water loss that does not involve diffusion. This was concluded to be water at the surface of the specimens, and was termed "superficial water". As such, it represents a fraction of the previously identified unbound (loose) water. Superficial water levels were: Chemflex 0.56%, Fuji IX 0.23%, Aquacem 0.87%. Equilibrium mass loss values were shown to be unaffected by temperature, and allowed ratios of bound:unbound water to be determined for all three cements. These showed wide variation, ranging from 1:5.26 for Chemflex to 1:1.25 for Fuji IX.


[Immunosuppressive treatments: mechanisms of action and clinical use].
Renal transplantation is the treatment of choice of end stage renal failure. It both improves the quality and the quantity of life compared to other techniques, such as hemodialysis. These results are partly related to the use of immunosuppressive therapy more effective and whose handling has improved over time. Advances in understanding the mechanisms of lymphocyte activation and the phenomena of rejection have in fact better defined the use of these treatments and their associations. Treatments can be broadly classified according to their characteristics (biological or chemical). Among chemical treatments, steroids are widely used, although the question of their avoidance or spearing is still a matter of debate. The cornerstone of immunosuppressive regimens remains the calcineurin inhibitors, characterized by a narrow therapeutic index and the need for therapeutic drug monitoring. Inhibitors of mammalian target of rapamycin (mTOR) have interesting antiproliferative effects that could be important against chronic allograft dysfunction and/or carcinogenesis. However, their safety profile makes them difficult to handle. Inhibitors of purine synthesis are largely based on inhibitors of inosine monophosphate dehydrogenase (IMPDH). Their effectiveness makes them privileged partners of other therapeutic classes. Among biological treatments, it is possible to separate the depleting and non depleting antibodies. Among the former, antithymocyte globulins are mainly active in T cells, whereas rituximab, a monoclonal anti-CD20, is active in B cells involved in the phenomena of humoral rejection. The non depleting antibodies are represented by anti-CD25, directed against the receptor for interleukin-2. In the near future it is likely that the belatacept, a costimulation blockade fusion protein will be used to allow calcineurin inhibitors sparing. Other immunosuppressive agents, acting at different levels of the immune response are being evaluated. In addition, advances in pharmacology offered hope of a better individualization of immunosuppressive therapies and better definition of therapeutic strategies used.


Ethnopharmacological approaches to Talinum paniculatum (Jacq.) Gaertn. - Exploring cardiorenal effects from the Brazilian Cerrado.
Talinum paniculatum (Jacq.) Gaertn. (Talinaceae), popularly known as "major gomes" and "erva gorda", is a non-conventional food plant extensively distributed throughout the Brazilian territory. In Brazilian folk medicine, this species is used as aphrodisiac, to treat gastrointestinal problems, and as a cardioprotective agent. However, there are no reports in the literature proving its cardiovascular effects. To perform a whole-ethnopharmacological investigation of the cardiorenal properties of the ethanol soluble fraction from T. paniculatum (ESTP) in Wistar rats. First, plant samples were collected, properly identified and a morpho-anatomical characterization was carried out to provide quality control parameters. Then, ESTP was obtained and its chemical profile was determined by LC-DAD-MS. In addition, an acute toxicity assay was conducted in female Wistar rats in order to observe any toxic effects after one single administration. Finally, the diuretic and hypotensive potential of ESTP (30, 100 and 300 mg/kg) were investigated in male rats followed by the evaluation of its possible effects on peripheral vascular resistance. Chemical compounds identified from ESTP were chlorogenic acids, amino acids, nucleosides, O-glycosylated flavones and organic acids. No signs of toxicity as well as no changes in urine volume or electrolyte elimination were observed after ESTP acute treatment. On the other hand, prolonged treatment with all doses of ESTP significantly increased urine volume and electrolyte excretion (Na+, K+ and Cl-) without affecting blood pressure or heart rate. Apparently, these effects are involved with the activation of the small conductance calcium-activated potassium channels contributing to the increase of renal blood flow and glomerular filtration rate. Data presented show important information about the ethnomedicinal properties of T. paniculatum. In addition, the study presents the ESTP as a possible herbal medicine, especially when a sustained diuretic effect is required.


Prediction of Superhalogen-Stabilized Noble Gas Compounds.
The discovery of HArF has generated renewed interest in the chemistry of noble gases, particularly their hydrides. Though many weak complexes of noble gases bound by van der Waals interactions are known, the number of halogenated noble gas compounds, HNgX (Ng = noble gas; X = halogen), where the noble gas atom is chemically bound, is limited. These molecules are metastable, and their specialty is that there is substantial ionic bonding between the noble gas atom and the halogen atom. In this Letter, it is shown using density functional theory and second-order Møller-Plesset perturbation theory that by replacing the halogen atoms by superhalogens (Y), whose electron affinities are much larger than those of halogens, more ionic bonds between Ng and Y can be attained. Moreover, the superhalogen-containing noble gas hydrides, HNgY, are more stable compared to their halogenated counterparts.


Association of extracellular cleavage of E-cadherin mediated by MMP-7 with HGF-induced in vitro invasion in human stomach cancer cells.
Proteolytic shedding of the ectodomain of a variety of transmembrane proteins, including cell-to-cell adhesion molecules, has been observed in solid cancers. We have investigated whether extracellular cleavage of E-cadherin mediated by matrix metalloproteinase-7 (MMP-7) is involved in hepatocyte growth factor (HGF) induced in vitro invasion in stomach cancer cells. The effects of HGF on the expression of E-cadherin/beta-catenin and MMP-7 at both the protein and mRNA levels were assessed in stomach cancer cells, NUGC-3 and MKN-28, and in cells in which the expression of MMP-7 was downregulated by transfection with a MMP-7 short hairpin RNA plasmid. Treatment with HGF increased the extracellular cleavage of E-cadherin and the release of MMP-7 and reduced the level of E-cadherin in a dose- and time-dependent manner. HGF treatment repressed the phosphorylation of beta-catenin in a Triton-soluble fraction, but enhanced this phosphorylation in a Triton-insoluble fraction. The association of E-cadherin with beta-catenin was decreased by HGF treatment in the Triton-soluble fraction. In addition, treatment of MMP-7 short hairpin RNA transfected NUGC-3 cells with HGF resulted in no extracellular cleavage of E-cadherin and also decreased the in vitro cell invasion. These results suggest that incubation with HGF mediated the release of MMP-7, resulting in extracellular cleavage of E-cadherin from stomach cancer cells. This might be a key mechanism in HGF-induced in vitro invasion and metastasis.


Outcome of patients with serology suggestive of past hepatitis B virus infection during antitumor necrosis factor therapy for psoriasis.
Recently, the reactivation during treatment with tumor necrosis factor (TNF) blockers has exceptionally been described in patients with hepatitis B virus (HBV) antigen-negative (HBsAg). The objective was to evaluate the influence of anti-TNF agents in patients with psoriasis and serology suggesting past hepatitis B state. The inclusion criteria were chronic plaque psoriasis treated with anti-TNF therapy, HBsAg-negative, and HBcAb-positive. We gathered the demographic data and type and duration of anti-TNF agent. Serum aminotransferase levels and HBV serologic status were requested at baseline and during follow-up. We have included 13 patients (four women, nine men) (mean age of 62.1 years). The agent was etanercept in seven cases, infliximab in four patients, and adalimumab in the other two. The mean duration of TNF therapy was 28.6 months. None of them became HBsAg-positive. Neither signs nor symptoms of acute hepatitis were reported. The management of HBsAg-negative patients is unresolved. Only nine cases of HBV reactivation during treatment with TNF blockers have been reported. Despite the low risk of reactivation in these patients, we recommend the monitoring of serum aminotransferase levels, HBsAb titers, HBsAg and, if possible, viral load.


Severe acute respiratory syndrome: 11 years later--a radiology perspective.
Severe acute respiratory syndrome (SARS) was a highly virulent atypical pneumonia caused by a novel coronavirus that resulted in a pandemic in 2003. Singapore was one of the most severely affected countries, and SARS took a heavy toll on our health care system. The lessons learned during the pandemic have shaped our national contagion response plan and have proved valuable in subsequent epidemics. We describe the lessons learned for the radiology department. Our experience with SARS has shaped and changed our daily practice of radiology.


Expanding the scope of aminosugars: synthesis of 2-amino septanosyl glycoconjugates using septanosyl fluoride donors.
A general strategy amenable to the strerocontrolled synthesis of complex, ring-expanded analogues of natural aminoglycosides has been developed. Central to the method is the utilization of septanosyl fluorides as glycosyl donors in facile and selective glycosylation reactions. The septanosyl fluorides proved to be the best choice for the glycosylations because of their accessibility and the scope of aglycones that they could glycosylate. Moreover, a high degree of stereoselectivity was observed in the glycosylations, exclusively giving 1,2-trans-glycosides. 2-Amino septanosyl fluorides were prepared from D-glucose, D-galactose, and D-mannose. Other routes to the septanosyl glyconjugates, especially with regard to alternate donor types, were systematically investigated. Since routes to the individual donor types were being explored, factors that exert a controlling influence on the acid-mediated cyclization of 1,6-hydroxy-aldehydes were determined. The newly prepared 2-amino septanosyl glycoconjugates illustrate the scope of the reaction and how it may be utilized for the preparation of other ring-expanded analogues of glycosylated natural products.


Postnatal development and differentiation of the secretory elements of the rabbit parotid and submandibular glands.
The development of the secretory elements of the parotid and submandibular glands has been examined in postnatal rabbits. In the submandibular gland, there are 2 principal types of secretory cells, serous and mucous. At birth, these occur in approximately equal proportions, and, by 28 days postnatal, the mucous elements greatly outnumber the serous ones. Initially, development of the parotid gland lags significantly behind that of the submandibular gland. Lobules, which are widely separated at birth, appear compact by 28 days postnatal, at which time also serous acinar cells contain an adult content of secretory granules. In both glands, secretory granules develop within component cells of intercalated ducts by 14 days postnatal. Although the glands appear virtually mature by 28 days postnatal, which corresponds approximately to the onset of weaning, minor structural changes occur over the following 3 postnatal weeks.


Cross-sectional left ventricular outflow tract velocities before and after aortic valve replacement: a comparative study with two-dimensional Doppler ultrasound.
To assess whether aortic valve replacement (AVR) results in changes in the flow velocity distribution in the left ventricular outflow tract (LVOT), 10 patients undergoing AVR for aortic stenosis were studied. By extracting velocity information from color flow maps as digital data, instantaneous cross-sectional velocity profiles were constructed. Velocity profiles obtained 1 to 3 days before AVR were compared with recordings made 3 months later. The LVOT velocity profiles were variably skewed both before and after surgery, and no systematic or uniform changes could be detected after AVR. The highest velocities were most often localized in the region from the center of the outflow tract diameter toward the septum both before and after surgery. At the time of peak flow the ratio of the maximum to the cross-sectional mean velocity was 1.38 +/- 0.13 before and 1.39 +/- 0.08 after AVR (NS), and the ratio of the maximum to the mean velocity time integral was 1.47 +/- 0.10 before and 1.56 +/- 0.10 after (NS). We conclude that AVR in patients with aortic stenosis does not result in a change in LVOT velocity profiles that will influence stroke volume estimates with the Doppler technique.


Reactive oxygen species and DNA damage after ultrasound exposure.
The aim of this work was to detect the formation of hydrogen peroxide and hydroxyl radicals after ultrasound (US) exposure and test the hypothesis that reactive oxygen species induced by ultrasound can contribute to DNA damage. Formation of reactive oxygen species was observed in incubated medium after sonication with 1 MHz continuous ultrasound at the intensities of 0.61-2.44 W/cm2. Free radicals and hydrogen peroxide produced by ultrasound exposure of cells can lead to DNA damage. Comet assay was used to assess the effect of ultrasound on the level of nuclear DNA damage. The nucleated erythrocytes from fish were exposed in vitro to ultrasound at the same intensities and frequency. It was noticed that ultrasound in all used intensities induced DNA damage. The effect was not eliminated by the addition of catalase, which indicates that DNA damage was not caused by hydrogen peroxide only. The results showed that the DNA damage can be repair and this mechanism was the most effective after 30 and 60 min after sonication. Furthermore, the ultrasound-induced DNA damage in the presence of sonosensitizer (Zn- and AlCl-phthalocyanine) was studied. It was noticed that phthalocyaniens (Pcs) alone or with ultrasound did not induce significant changes in the level of DNA damage.


Risk of metastatic pheochromocytoma and paraganglioma in SDHx mutation carriers: a systematic review and updated meta-analysis.
Pheochromocytoma and paraganglioma (PPGL) are tumours that arise from chromaffin cells. Some genetic mutations influence PPGL, among which, those in genes encoding subunits of succinate dehydrogenase (SDHA, SDHB, SDHC and SDHD) and assembly factor (SDHAF2) are the most relevant. However, the risk of metastasis posed by these mutations is not reported except for SDHB and SDHD mutations. This study aimed to update the metastatic risks, considering prevalence and incidence of each SDHx mutation, which were dealt formerly all together. We searched EMBASE and MEDLINE and selected 27 articles. The patients included in the studies were divided into three groups depending on the presence of PPGL. We checked the heterogeneity between studies and performed a meta-analysis using Hartung-Knapp-Sidik-Jonkman method based on a random effect model. The highest PPGL prevalence was for SDHB mutation, ranging from 23% to 31%, and for SDHC mutation (23%), followed by that for SDHA mutation (16%). The lowest prevalence was for SDHD mutation, ranging from 6% to 8%. SDHAF2 mutation showed no metastatic events. The PPGL incidence showed a tendency similar to that of its prevalence with the highest risk of metastasis posed by SDHB mutation (12%-41%) and the lowest risk by SDHD mutation (~4%). There was no integrated evidence of how SDHx mutations are related to metastatic PPGL. However, these findings suggest that SDHA, SDHB and SDHC mutations are highly associated and should be tested as indicators of metastasis in patients with PPGL.


Potential of GPCRs to modulate MAPK and mTOR pathways in Alzheimer's disease.
Despite efforts to understand the mechanism of neuronal cell death, finding effective therapies for neurodegenerative diseases is still a challenge. Cognitive deficits are often associated with neurodegenerative diseases. Remarkably, in the absence of consensus biomarkers, diagnosis of diseases such as Alzheimer's still relies on cognitive tests. Unfortunately, all efforts to translate findings in animal models to the patients have been unsuccessful. Alzheimer's disease may be addressed from two different points of view, neuroprotection or cognitive enhancement. Based on recent data, the mammalian target of rapamycin (mTOR) pathway arises as a versatile player whose modulation may impact on mechanisms of both neuroprotection and cognition. Whereas direct targeting of mTOR does not seem to constitute a convenient approach in drug discovery, its indirect modulation by other signaling pathways seems promising. In fact, G-protein-coupled receptors (GPCRs) remain the most common 'druggable' targets and as such pharmacological manipulation of GPCRs with selective ligands may modulate phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), mitogen-activated protein (MAP) kinase and mTOR signaling pathways. Thus, GPCRs become important targets for potential drug treatments in different neurodegenerative disorders including, but not limited to, Alzheimer's disease. GPCR-mediated modulation of mTOR may take advantage of different GPCRs coupled to different G-dependent and G-independent signal transduction routes, of functional selectivity and/or of biased agonism. Signals mediated by GPCRs may act as coincidence detectors to achieve different benefits in different stages of the neurodegenerative disease.


Immunisation status of children in the eastern Bay of Plenty.
To assess the status of immunisation of children in the eastern Bay of Plenty. Survey of general practitioner records and case notes of all resident children born over a period of two years and aged from five to 29 months. Immunisation was considered up to date if received within a reasonable period of the expected time. One thousand five hundred and ninety-three children were studied (97% of those known to be resident). The six week immunisations were received by 91%, 88% received the three month, and 81% the five month vaccines. Delivery of second year immunisation was much less successful, only 72% receiving measles or MMR vaccine despite publicity about a current measles epidemic and 61% the eighteen month vaccines. Forty-three percent children of two years of age had not received the full recommended immunisation schedule. Some of the contributing factors were identified and measures that could improve immunisation uptake are discussed.


Sickness absence and psychosocial work conditions: a multilevel study.
Psychosocial work conditions, particularly psychological job demands, are inconsistently associated with sickness absence rates. This might be the result of investigating the psychosocial work environment at the individual level, reflecting personal perceptions rather than actual demands. To investigate associations between sickness absence and psychosocial work conditions at both the individual and the workplace level. A cross-sectional study of insurance company employees (n = 395) in four departments. Psychological job demands, job control and job support were investigated at the individual level using the self-completed Questionnaire on Experience and Assessment of Work. An external occupational psychologist interviewed the supervisor and a group of employees of each department, assessing job demands, job control, job support and psychological distress at the workplace level. These data were related to the number of short (1-7 days), medium (8-21 days) and long (>21 days) episodes of sickness absence in the period January 2001 to December 2002. A total of 244 questionnaires (62%) were suitable for analysis. Quantitative job control scores at the individual level differed from qualitative data at the workplace level. Self-assessed job demands and control were unrelated to sickness absence. The rates of short and long episodes of absence were higher in the department with combined high demands and low control, assessed at the workplace level. The associations between psychosocial work conditions and sickness absence depended on the level at which the former were assessed. More multilevel research is needed to disentangle the relations between psychosocial work conditions and sickness absence.


Comparison of statistical significance criteria.
We study and compare two classes of statistical criteria to assess the significance of exceptional words. Indeed, the Z-score-like criteria, or the normal approximation that is a strict equivalent, suffer from several drawbacks in terms of sensitivity and specificity. Thanks to the combinatorial structure of words, a computation of the exact P-value has been made possible by recent mathematical results. We study here the drawbacks of the Z-score, the choice of the threshold and the tightness to the P-value. A major conclusion is that the normal approximation is always very poor and overestimates statistical significance.


Oxidative ring cleavage of cyclic acetals with hypervalent tert-butylperoxy-lambda(3)-iodanes.
[reaction: see text] Exposure of cyclic acetals to 1-tert-butylperoxy-1,2-benziodoxol-3(1H)-one in the presence of tert-butyl hydroperoxide and potassium carbonate in benzene at room temperature results in oxidative ring cleavage to glycol monoesters via intermediate tert-butylperoxy ortho esters.


Combined effects in toxicology--a rapid systematic testing procedure: cadmium, mercury, and lead.
A testing procedure is described for the assessment of the toxicological response (e.g., acute toxicity or mutagenicity) of any combination and number of chemical, physical, and biological agents, with no more effort for a particular combination than for a single agent. The method provides a simple, sensitive, and quantitative index of synergism, antagonism, and additivity, and it has been demonstrated experimentally in rats by determining the acute lethality of combinations of cadmium, mercury, and lead salts. In a combination of two metal salts, the dose of one metal of the pair was fixed at or near the no-effect level while the dose of the second metal was increased until the entire dose-response curve was obtained. To evaluate interactions of the three metals, the previous pair of metals were kept fixed at their combined extrapolated LD1 level, and the third metal was increased. The statistical treatment of the data employed a computer program that did not involve probit transformations, but rather the approximate linear relationship between the fractional response and the logarithm of the dose. A particular combination could be synergistic, antagonistic, or additive, depending on the relative doses employed. Generally, a combination was synergistic when the most toxic member was present at or near its LD1 dose in the presence of the much less toxic member; the same combination was protective when the least toxic member was present at or near its LD1 dose. The results clarify apparently contradictory reports regarding the biological effects of metal combinations. The application of the testing procedure to combinations of mutagens is described, and an example is cited involving, for a particular bacterial mutagen, a combination of N-methyl-N'-nitro-N-nitrosoguanidine with ethylmethanesulfonate.


The attentional demands of negation in a memory-scanning task.
Subjects were run in a Sternberg recognition memory task modified to occasionally require a transformation prior to response. The transformation was similar to logical negation since onplus or nontransformed trials subjects responded "yes" if the probe was from the current memory set and "no" otherwise, while onminus or transformed trials subjects responded "no" to memory set probes and "yes" to nonmemory set probes. Two models proposed to account for the effects of logical negation were compared: The encoding and comparison model predicts that logical negation requires an additional processing stage, while the capacity sharing model argues that negation adds a processing stage and also retards the rate of memory search. The results indicated that minus probes had 50% larger mean reaction time by set size slopes than the corresponding plus probes. This finding offers clear support for the capacity sharing model.


Facial asymmetry in superior oblique muscle palsy.
Facial asymmetry is an underrecognized association or sequela of torticollis in congenital or very early onset superior oblique muscle palsy, which when present in an adult, helps to confirm chronicity and prevent unnecessary neurologic evaluation. Nineteen cases of superior oblique muscle palsy were studied prospectively. Of 12 unilateral palsies presenting in adulthood, 9 were considered congenital by history and 7 (77%) of these had facial asymmetry. Large vertical fusional vergence amplitudes and torticollis, on review of childhood photographs, confirmed the early onset of the palsy in each facial asymmetry patient. In addition, two children presenting at age 18 months and 3 years of age with constant head tilting had facial asymmetry. If torticollis is corrected before maturation of the facial structure, resolution of the asymmetry may occur.


The older sickle cell patient.
The lifespan of sickle cell patients has increased substantially. We chose to compare the attributes of older sickle cell patients with younger ones within the confines of a single institutional study serving the local Bronx population. Laboratory and clinical assessments from 40 patients with sickle cell disease over 40 years of age were compared to 40 patients under 30 years of age. When the older group was compared to the younger group, hemoglobin, indirect bilirubin, and platelet counts were significantly lower. Creatinine clearances were lower and BUN levels were higher in the older group. Although the prevalence of radiographically measured cardiomegaly was significantly higher in the older group, systolic and diastolic blood pressures remained unchanged. The number of crises and number of admissions were higher in the younger group than in the older, but there was wide variation, and no significant differences could be detected; days spent in hospital over the previous 2 years did not differ. No statistically significant differences were observed for absolute reticulocyte count, MCV, % HbF, white cell count, liver function tests, or oxygen saturation. Comparing genders within age groups, younger males had higher hemoglobin levels and absolute reticulocyte counts than younger females, while only the older females had higher HbF levels than the corresponding males. Lower levels of hemoglobin associated with older age may reflect decreased hematopoietic potential that may in part be due to decreased renal function. Lower platelet counts in older sickle cell patients may be a consequence of this decreased hematopoietic potential but may also represent a secondary survival benefit effect in sickle cell anemia.


In vitro and in vivo association of dentin phosphophoryn with alpha1CB6 peptide of type I collagen.
A small number of molecules of phosphophoryns, dentin phosphoproteins, are associated to collagen tightly with the maturation of dentin. As an in vitro model of the process, we examined non-covalent association of phosphophoryns and collagen CNBr peptides. Cyanogen bromide peptides of type I collagen were separated with electrophoresis and transferred to a membrane, which was probed with labeled phosphophoryns. Phosphophoryns were bound preferentially to alpha1CB6 peptide. Another experiment using a cleavable crosslinking agent also demonstrated the affinity between phosphophoryns and the alpha1CB6 region of collagen. The matrix-bound fraction of phosphophoryns was solubilized by CNBr digestion of bovine dentin, and was partially purified. Compositional analysis revealed that the fraction was composed of association products of phosphophoryns and collagen at the ratio of 2:3. Considering the high molecular weight of the fraction, one phosphophoryn molecule should be associated with several collagen peptides. The fraction reacted with the antibody against alphalCB6 peptide. When the fraction was digested with lysyl endopeptidase, several peptides that coincided with peptides from alpha1CB6 were released. One of the peptides was sequenced and identified to be a peptide with Asp 975 of the alpha1(I) chain as an amino terminal residue. The alpha1CB6 peptide may be involved in the matrix-bound fraction, although involvement of other collagen peptides cannot be ruled out. Phosphophoryns may associate with collagen at the alpha1CB6 region at first, and then be immobilized on it, possibly by covalent crosslinking, with maturation of dentin. Binding of several alpha1CB6 peptides to a single phosphophoryn molecule is possible in current 3-dimensional models of collagen packing in mineralized tissue, which allows alignment of several hole zones in register.


Inhibition of cytotoxic activity of carp lymphocytes (Cyprinus carpio) by anti-thymocyte monoclonal antibodies.
A series of monoclonal antibodies (mAbs) were developed against carp thymocytes and carp immunoglobulin (IgM). Anti-thymocyte mAbs reacted with thymocytes, head kidney lymphocytes (HKL) as well as peripheral blood lymphocytes (PBL) at various proportions. Anti-IgM antibody reacted with HKL and PBL but not with thymocytes. PBL from carp immunized with xenogeneic mouse myeloma cells exhibited cytotoxic activity against target cells but PBL from unimmunized fish showed weak cytotoxicity. Treatment of PBL with anti-thymocyte mAbs and guinea pig complement suppressed the cytotoxic activity dose dependently; however, anti-IgM mAb did not. These results indicate the cytotoxic cells in PBL possess cell surface antigens common to thymocytes.


Döderlein-Krönig vaginal hysterectomy: an alternative to the traditional Heaney hysterectomy.
The aim of this study was to demonstrate the surgical steps of performing the Döderlein-Krönig hysterectomy. A video recording was made of the hysterectomy using an alternative technique described by Drs. Döderlein and Krönig in 1906. The patient in the video had stage 3 prolapse and desired surgical correction. The video demonstrates an exam under anesthesia, an anterior colpotomy, delivery of the uterine fundus through the colpotomy, and completion of the hysterectomy. The surgery was performed by a urogynecologist and a resident physician. The patient tolerated the procedure well and had no symptom recurrence at her 1-year follow-up visit. This video was presented at the 2014 International Urogynecological Association Annual Meeting in Washington, DC, as a nondiscussed video poster. This video can assist and educate others in using this technique for performing a hysterectomy. Possible benefits of this approach include decreased blood loss and improved visualization, especially in women with pelvic organ prolapse.


Closed-loop glycaemic control using an implantable artificial pancreas in diabetic domestic pig (Sus scrofa domesticus).
The performance of a completely implantable peritoneal artificial pancreas (AP) has been demonstrated in principle in a live diabetic domestic pig. The device consists of a smart glucose-sensitive gel that forms a gateway to an insulin reservoir and is designed to both sense glucose and deliver insulin in the peritoneal cavity. It can be refilled with insulin via subcutaneous ports and surgery was developed to insert the AP. Diabetes was induced with streptozotocin (STZ), the device filled with insulin (Humulin(®) R U-500) in situ and the animal observed for several weeks, during which time there was normal access to food and water and several oral glucose challenges. Blood glucose (BG) levels were brought down from >30 mmol/L (540 mg/dL) to non-fasted values between 7 and 13 mmol/L (126-234 mg/dL) about five days after filling the device. Glucose challenge responses improved ultimately so that, starting at 10 mmol/L (180 mg/dL), the BG peak was 18 mmol/L (324 mg/dL) and fell to 7 mmol/L (126 mg/dL) after 30 min, contrasting with intravenous attempts. The reservoir solution was removed after 8 days of blood glucose levels during which they had been increasingly better controlled. A rapid return to diabetic BG levels (30 mmol/L) occurred only after a further 24 days implying some insulin had remained in the device after removal of the reservoir solution. Thus, the closed loop system appeared to have particular influence on the basal and bolus needs for the 8 days in which the reservoir solution was in place and substantial impact for a further 3 weeks. No additional insulin manual adjustment was given during this period.


Role of expiratory flow limitation in determining lung volumes and ventilation during exercise.
We determined the role of expiratory flow limitation (EFL) on the ventilatory response to heavy exercise in six trained male cyclists [maximal O2 uptake = 65 +/- 8 (range 55-74) ml. kg-1. min-1] with normal lung function. Each subject completed four progressive cycle ergometer tests to exhaustion in random order: two trials while breathing N2O2 (26% O2-balance N2), one with and one without added dead space, and two trials while breathing HeO2 (26% O2-balance He), one with and one without added dead space. EFL was defined by the proximity of the tidal to the maximal flow-volume loop. With N2O2 during heavy and maximal exercise, 1) EFL was present in all six subjects during heavy [19 +/- 2% of tidal volume (VT) intersected the maximal flow-volume loop] and maximal exercise (43 +/- 8% of VT), 2) the slopes of the ventilation (DeltaVE) and peak esophageal pressure responses to added dead space (e.g., DeltaVE/DeltaPETCO2, where PETCO2 is end-tidal PCO2) were reduced relative to submaximal exercise, 3) end-expiratory lung volume (EELV) increased and end-inspiratory lung volume reached a plateau at 88-91% of total lung capacity, and 4) VT reached a plateau and then fell as work rate increased. With HeO2 (compared with N2O2) breathing during heavy and maximal exercise, 1) HeO2 increased maximal flow rates (from 20 to 38%) throughout the range of vital capacity, which reduced EFL in all subjects during tidal breathing, 2) the gains of the ventilatory and inspiratory esophageal pressure responses to added dead space increased over those during room air breathing and were similar at all exercise intensities, 3) EELV was lower and end-inspiratory lung volume remained near 90% of total lung capacity, and 4) VT was increased relative to room air breathing. We conclude that EFL or even impending EFL during heavy and maximal exercise and with added dead space in fit subjects causes EELV to increase, reduces the VT, and constrains the increase in respiratory motor output and ventilation.


Intracellular signalling pathways induced by chemokines in natural killer cells.
Chemokines are small peptides involved in the recruitment of various cell types into inflammatory sites. They are divided into four sub-families depending on the presence of amino acids separating the cysteine residues in their N-terminal region. These are the alpha (CXC), beta (CC), gamma (C) and delta (CX)C) chemokines. In addition, five CXC chemokine (CXCR1-5), nine CC chemokine (CCR1-9), one C chemokine (XCR1) and one C-X3C chemokine (CX3CR1) receptors have been identified. These receptors belong to the seven transmembrane spanning domain family, and are coupled to the heterotrimeric guanine nucleotide binding (G) proteins. Chemokines activate various immune cells, and in particular the anti-viral/anti-tumour effectors, the natural killer (NK) cells by activating members of the heterotrimeric G proteins. The importance of the family of chemokines is highlighted by the ability of its members to inhibit the replication of HIV-1 strains in CD4+ cells, where chemokine receptors act as HIV-1 co-receptors. This review discusses the intracellular signalling pathways induced by chemokines in NK and other cell types, and the relationships to HIV-1 signalling in these cells.


[Germ cell tumor in adolescents and young adults].
Management of extracranial germ cell tumors (TG) is emblematic of the adolescents and young adults' problematic. Beyond persisting differences in care between adult and pediatric oncologists, it emphasizes the need for effective collaboration between both teams. If the therapy is primarily guided today by adult standards, pediatricians bring expertise in long-term follow-up, justified by the increasing description of late side effects in this young population, with highly curable disease since cisplatin's emergence.


Relationship of adiponectin with metabolic syndrome components in pubertal children.
Adiponectin is an adipose tissue-derived adipocytokine which appears in decreased concentrations in obese patients and in several processes related to cardiovascular disease, such as type 2 diabetes or metabolic syndrome. The aim of this study was to analyze the relationship between adiponectin and components of metabolic syndrome (lipid profile, blood pressure, insulin and insulin resistance) in pubertal Spanish children. The population-based sample included 810 healthy children (382 boys and 428 girls) 12-16 years of age. Anthropometric parameters and blood pressure were measured. Lipid levels were determined by standard methods, and insulin and adiponectin concentrations were measured by ELISA. Insulin resistance index was assessed by HOMA-IR. Adiponectin levels were negatively correlated with insulin and HOMA in both boys and girls, and remained significant after adjustment for BMI z-score in girls. After this adjustment, adiponectin maintained a positive correlation with HDL-cholesterol and HDL-phospholipids in both genders, and correlated with triglycerides in girls. Multiple linear regression analysis showed that, after adjustment for BMI z-score, adiponectin accounted for 15.8% of the variation of HDL-cholesterol in girls and for 5% of its variation in boys; meanwhile, it accounted for 15.8% and 12.7% of the variation of HDL-phospholipids in girls and boys, respectively. Adiponectin levels in 12- to 16-year-old children appear to be more strongly related to better lipid profile and insulin sensitivity in girls than in boys. Our study shows, for the first time to our knowledge, a significant positive correlation between adiponectin and HDL-phospholipids in pubertal children.


Lymphocyte subpopulations in allergic granulomatosis and angiitis (Churg-Strauss syndrome).
We report a case of allergic granulomatosis and angiitis (Churg-Strauss syndrome) in which immunologic parameters, including lymphocyte subpopulations, were determined in the acute phase of the disease and during remission. Hyperimmunoglobulinemia E and immune complexes were present. A low proportion of suppressor/cytotoxic (T8+) lymphocytes and a high helper/suppressor ratio were seen throughout the course of the disease, although immunoglobulin levels and circulating immune complex levels decreased with therapy. We hypothesize that the deficiency of suppressor cells may play a role in the pathogenesis of this syndrome.


Reaction-path calculations and crystal structures of 1,1'-(ethylene-1,2-diyl)dipyridinium dichloride dihydrate and 1,1'-(ethylene-1,2-diyl)dipyridinium dibromide.
The design of new organic-inorganic hybrid ionic materials is of interest for various applications, particularly in the areas of crystal engineering, supramolecular chemistry and materials science. The monohalogenated intermediates 1-(2-chloroethyl)pyridinium chloride, C5H5NCH2CH2Cl(+)·Cl(-), (I'), and 1-(2-bromoethyl)pyridinium bromide, C5H5NCH2CH2Br(+)·Br(-), (II'), and the ionic disubstituted products 1,1'-(ethylene-1,2-diyl)dipyridinium dichloride dihydrate, C12H14N2(2+)·2Cl(-)·2H2O, (I), and 1,1'-(ethylene-1,2-diyl)dipyridinium dibromide, C12H14N2(2+)·2Br(-), (II), have been isolated as powders from the reactions of pyridine with the appropriate 1,2-dihaloethanes. The monohalogenated intermediates (I') and (II') were characterized by multinuclear NMR spectroscopy, while (I) and (II) were structurally characterized using powder X-ray diffraction. Both (I) and (II) crystallize with half the empirical formula in the asymmetric unit in the triclinic space group P-1. The organic 1,1'-(ethylene-1,2-diyl)dipyridinium dications, which display approximate C2h symmetry in both structures, are situated on inversion centres. The components in (I) are linked via intermolecular O-H...Cl, C-H...Cl and C-H...O hydrogen bonds into a three-dimensional framework, while for (II), they are connected via weak intermolecular C-H...Br hydrogen bonds into one-dimensional chains in the [110] direction. The nucleophilic substitution reactions of 1,2-dichloroethane and 1,2-dibromoethane with pyridine have been investigated by ab initio quantum chemical calculations using the 6-31G** basis. In both cases, the reactions occur in two exothermic stages involving consecutive SN2 nucleophilic substitutions. The isolation of the monosubstituted intermediate in each case is strong evidence that the second step is not fast relative to the first.


Comparison of habit reversal and a behaviorally-modified dental treatment for temporomandibular disorders: a pilot investigation.
This study tested the hypothesis that a habit reversal program emphasizing awareness and reduction of masticatory muscle activity would significantly reduce pain in patients diagnosed with chronic temporomandibular disorder (TMD) and would be a competitive alternative to a behaviorally-modified dental intervention. Eight individuals diagnosed with TMD were randomly assigned to a splint therapy or habit reversal group. Patients in the splint group received an interocclusal appliance (splint) fabricated from acrylic and were instructed to wear the splint day and night up to a maximum of 20 h per day. Patients in the habit reversal group were given a pager and instructed to check tooth position and masticatory muscle tension when paged. Paging occurred approximately once every 2 h during the day, but not at night. Both groups were instructed to avoid tooth contact and relax the masticatory muscles during the 4 weeks of active treatment. Outcome data were collected at 1 month and 1 year post-treatment intervals. Pain decreased significantly for both groups and did not differ between groups. Habit reversal may be as effective as a behaviorally-modified splint therapy for TMD-related pain.


[Polymorphism of ISSR-PCR markers in Tuvinian population of reindeer Rangifer tarandus L].
The results of the first analysis of the reindeer population inhabiting the south of Siberia, the Republic of Tyva (Tuva), performed by use of inter-simple sequence repeat (ISSR) technique, are reported. Using ISSR-PCR with two primers, a total of 71 fragments of different size were detected. Furthermore, for the population examined the indices of the mean pairwise similarity and the mean heterozygosity were calculated. The approach used can be recommended as a useful tool for the genetic diversity monitoring in Tuvinian population of reindeer.


Experience with streptokinase treatment of major pulmonary embolism.
The efficacy of streptokinase was studied in an open trial on 30 consecutive patients with major pulmonary embolism, affecting at least three lobar arteries in pulmonary angiography. Streptokinase was started on average 26 hours from the onset of symptoms and usually given directly to the pulmonary artery. Conventional dosage was used and adjusted by thrombin time. The average treatment period was 32 hours. 26 patients (87%) survived and made a good clinical recovery. Improvement occurred early: signs of improved pulmonary perfusion appeared usually within four hours. Repeat angiograms in 20 patients showed remarkable increase in pulmonary perfusion and partial but not complete lysis of the emboli. Four patients (13%) died, three of them because of no response and one due to late cardiac arrhythmia. Bleeding requiring blood transfusion occurred in four patients, but caused no late sequelae. It is concluded that streptokinase treatment of major pulmonary embolism is effective and safe in rapidly restoring pulmonary perfusion.


Effect of a high dose of fluoride on resorbing osteoclasts in vivo.
Histomorphometric analysis of osteoclasts resorbing bone above the erupting first rat molar tooth germ was made at 1, 2, 6, 24, and 48 h following systemic administration of a single high dose of sodium fluoride. Compared to the number of osteoclasts in control rats, a significant reduction was observed in the number of actively resorbing osteoclasts in fluoride-injected rats at 2 and 6 h after dose administration, after which a gradual increase to control levels was observed. A locally increased number of inactive osteoclasts, not attached to bone surfaces, accompanied the reduced number of active osteoclasts.


In vitro proliferation and in vivo malignancy of cell lines simultaneously derived from a chemically-induced heterogeneous rat mammary tumor.
Identification of clones in primary tumors responsible for proliferation, invasion, and metastasis was carried out. Four different aneuploid established cell lines derived from a ductal infiltrating mammary rat tumor induced by 7,12-dimethylbenz[a]anthracene were studied for proliferative and growth features in vitro and for tumorigenic and metastatic potential in vivo in nude mice. Clones, named RM1, RM2, RM3, and RM4, were characterized by different proliferative activity. Clone RM1 showed the highest proliferative activity by both tritiated thymidine incorporation and S-phase flow cytometry, followed by clone RM4. Conversely, clones RM2 and RM3 showed a lower proliferation rate. Growth-promoting activity, tested on 3T3 Swiss cells, was high in all clones, although RM1 showed significantly lower growth factors-releasing activity. Nude mice tumorigenesis demonstrated a strong tumor induction of line RM1 (100% of the mice after 47 +/- 7 d) and a slightly lower tumor induction of line RM4 (70% of the mice after 69 +/- 9 d). Line RM3 showed tumor induction in 40% of the mice after 186 +/- 16 d. Lines RM2 showed no tumor induction. Metastasis occurred in mice treated with line RM1 only. Therefore, tumorigenesis and metastasis correlate with proliferation but not with the release of growth factors. In conclusion, flow cytometry monitoring of clones from heterogeneous primary tumors proved to be a suitable model for the study of in vivo malignancy and in vitro proliferation.


Enhancement of protective immune responses to Venezuelan equine encephalitis (VEE) virus with microencapsulated vaccine.
Venezuelan equine encephalomyelitis (VEE) virus is a mosquito-borne arbovirus of major human health significance in the New World. Currently two forms of VEE virus are used for immunization of humans and horses, i.e. a live attenuated and a formalin-inactivated vaccine. Clinical evidence suggests that these vaccines are not fully efficacious and may produce certain undesirable side-effects. In the present study, microspheres composed of biocompatible and biodegradable poly (DL-lactide-co-glycolide) (DL-PLG) were evaluated for their effectiveness as a delivery system of whole, inactivated VEE virus vaccine for the induction of protective immune responses. Mice receiving 50 micrograms VEE virus in microspheres composed of an equimolar ratio of DL-lactide and glycolide (50:50 DL-PLG) exhibited a primary circulating IgG antibody response which was approximately 32-times higher than the response induced with the same dose of unencapsulated (free) virus. A similar difference in responses was seen with antigen doses ranging from 3.1 to 50 micrograms. A rapid increase in antibody activity was seen after the secondary immunization (day 50). Formalin fixation of inactivated VEE virus was important for immunogenicity since the circulating anti-VEE virus antibody response induced with microencapsulated nonformalin-fixed virus vaccine was lower than that induced with microencapsulated formalin-fixed virus vaccine. Furthermore, at low antigen concentrations, DL-PLG microsphere vaccines prepared with the solvent methylene chloride induced higher antibody responses than those prepared using ethyl acetate as the solvent. Microencapsulated vaccine also induced higher VEE virus neutralization titers than did free virus vaccine. Finally, the microencapsulated virus was more effective than the free virus in inducing immune responses protective against systemic challenge with virulent VEE virus. These results demonstrate that DL-PLG microspheres containing formalin-fixed, inactivated VEE virus were effective in augmenting circulating IgG antibody levels and neutralization titers to the VEE virus following systemic immunization and in affording enhanced protection against systemic challenge with virulent VEE virus. The effects of antigen form and the microsphere processing solvent on the immunogenicity of the vaccine are discussed.


Yeast Kre1p is a cell surface O-glycoprotein.
The Saccharomyces cerevisiae KRE1 gene encodes a secretory protein required for the production of the cell wall polymer (1-->6)-beta-glucan. Here we report further characterization of the KRE1 gene product, Kre1p. A functional, epitope-tagged Kre1p is shown to be highly modified in a SEC53-dependent manner. Kre1p is O-glycosylated, but the basis for the majority of its post-translational modification is unknown. Fractionation of Kre1p reveals a cell wall-associated form and a less abundant membrane-associated species. Indirect immunoflurorescence demonstrates that Kre1p localizes to the cell surface, where it becomes concentrated at the surface of mother cells. Such a localization of Kre1p seems to parallel the CAL1/CSD2-dependent cell wall deposition of chitin found in S. cerevisiae, and is consistent with evidence from Schizophyllum commune that (1-->6)-beta-glucan accumulates during maturation of the subapical region of the wall distal to the hyphal tip.


m7FLIPI and targeted sequencing in high-risk follicular lymphoma.
Patients with follicular lymphoma (FL) refractory to front-line immunochemotherapy (ICT) have a poor overall survival (OS). Gene mutation analysis may be more accurate than classical risk factors to pick out these patients before treatment. This study aimed to describe the prevalence of selected genetic mutations in a cohort of patients with high-risk FL. Twenty-five patients with FL refractory to front-line ICT and 10 non-refractory patients matched for age, sex, and FLIPI score were included. We sequenced 18 genes (custom targeted sequencing panel) previously reported to potentially have prognostic impact, including the seven genes necessary to determine m7FLIPI risk. The 35 patients had a median age of 62. The FLIPI and FLIPI2 were high in 27 (84%) and 14 (48%), respectively. Three-year progression-free survival (PFS) and OS probabilities were 25% (95% CI, 13%-41%) and 53% (34%-69%), respectively. There were 73 variants in the 18 genes among the 35 patients. The median number of mutations per patient was 1 (interquartile range, 0-3). The most commonly mutated genes were CREBBP (11 of 35, 31%) and EP300 (10 of 35, 29%). EP300 mutations were associated with refractoriness to treatment (10 of 25 among refractory and 0 of 10 among non-refractory). In conclusion, in this study, patients with high-risk follicular lymphoma were genetically heterogeneous.


[The mini-clinical evaluation exercise: from 'judgement' to 'guidance'].
The mini-clinical evaluation exercise (mini-CEX) is a sort of test in which the assessor's evaluation is based on a short observation of the resident in a clinical situation. A mini-CEX is not an examination but a useful educational instrument to monitor and to foster the resident's development. The central concepts of a good mini-CEX are 'short' (limited both in content and duration) and 'often' (the more frequent the better). It is important the assessor should provide feedback as soon as possible after the evaluation; this feedback should be constructive and task-oriented and be presented to the resident in a credible manner. Multiple observations by different assessors will ensure that the overall picture of the resident's progress is more reliable. Because the particular value of the mini-CEX lies mainly in the repetition involved, we would like to replace the 'E' for 'evaluation' with the 'G' for 'guidance'.


Aldosterone and the control of lower intestinal Na+ absorption and Cl- secretion in chickens.
There are several ion-transport systems expressed in the lower intestinal segments of chickens, depending on the level of the salt intake. The aim of this study was to test the hypothesis that aldosterone is the sole regulator of all these ion-transport systems, as had been indicated by our previous results. Chickens were long-term adapted to low salt intake, and then switched to a high-salt diet. During the first 5 days of resalination, the birds were injected with aldosterone every 8 hr and then the magnitude and characteristics of the epithelial ion-transport systems in colon and coprodeum were investigated. The results support strongly the hypothesis that aldosterone exerts major control of the amiloride-inhibitable Na(+)-transport system in both colon and coprodeum, as its magnitude was maintained high in spite of the resalination process. Spironolactone counteracted the actions of aldosterone, although not totally. On the other hand, aldosterone is not the sole regulator of the hexose/aminoacid-Na+ cotransport systems in colon, although it can act as their modulator, as the injections did delay the normal increase always seen in these transport systems during resalination. Aldosterone can also modulate the Cl(-)-secretory capacity of colon and coprodeum, but only temporarily.


[Peritonitis in elderly patients. Diagnostic and therapeutic considerations].
Abdominal surgical emergencies in the aged patients are considered on the basis of personal experience. Peritonitis from acute infections, is for many reasons, a higher challenge in the elderly.


Second messenger enzymes in glial cells: a cytochemical point of view.
Knowledge about second messenger metabolizing enzymes in neuroglia is still rather fragmentary. Therefore, the aim of the present investigation was to localize adenylate cyclase, guanylate cyclase, cyclic nucleotide phosphodiesterase and protein kinase A in glial cells of the rat hippocampus and cerebellum. Enzyme histochemical and immunohistochemical methods were used to detect the enzymes at the light and electron microscopic level. Astroglial cells were found to contain all 4 enzymes. Especially the microvascular glial cell processes were reactive. Oligodendroglial cells were only stained for adenylate cyclase acticity. Intracellularly, microtubules and intracellular membranes were frequently stained. The results point to the regulation of glial cell metabolism and of transport processes by cyclic nucleotides.


Interaction effect of PGC-1α rs10517030 variants and energy intake in the risk of type 2 diabetes in middle-aged adults.
PGC-1α is an important regulatory factor for energy and glucose metabolism. Therefore, we investigated whether the PGC-1α genotype (rs10517030 and rs10212638) affects the incidence of type 2 diabetes mellitus (T2DM) and sought to explain the interactions between their variants and nutrient intake on the development of T2DM. Subjects aged 40-65 years of both genders were from the Ansung/Ansan cohorts (8842 adults) in Korea. Associations of PGC-1α variants rs10517030 and rs10212638 with T2DM were analyzed in a dominant genetic model, and were tested for interactions of genotypes and nutrients with T2DM risk. It was adjusted for covariates related to glucose metabolism. Three variants, rs10517030, rs10517032 and rs10212638, were positively associated with T2DM prevalence. Single-nucleotide polymorphisms, rs10517030 and rs10517032, had strong association (r2=0.963). In the glucose tolerance tests, odds ratios (ORs) for serum glucose levels at 120 min were higher for subjects who were in the minor-allele group (minor allele homozygotes and heterozygotes) than for the major-allele group (major allele homozygotes) for rs10517030 variants. Serum insulin levels at 60 min had a lower ORs in the minor-allele group of rs10517030 variants. The interaction between energy intake and PGC-1α rs10517030 variants also affected T2DM risk. PGC-1α minor alleles were linked to T2DM prevalence and homeostasis model assessment estimate of insulin resistance (HOMA-IR) only in the low-energy groups, and HOMA-B was significantly negatively associated with the minor-allele group of PGC-1α rs10517030 variants, only in the low-energy-intake groups. These data suggest that Koreans with the minor alleles of PGC-1α rs10517030, rs10517032 and rs10212638 are at greater risk of T2DM, and that a low-energy diet is more protective against the development of T2DM in subjects with the major alleles of rs10517030 and rs10517032.


Heterochromatin and histone modifications in the germline-restricted chromosome of the zebra finch undergoing elimination during spermatogenesis.
In the zebra finch (Taeniopygia guttata) a germline-restricted chromosome (GRC) is regularly present in males and females. While the GRC is euchromatic in oocytes, in spermatocytes this chromosome is cytologically seen as entirely heterochromatic and presumably inactive. At the end of male meiosis, the GRC is eliminated from the nucleus. By immunofluorescence on microspreads, we investigated HP1 proteins and histone modifications throughout male meiotic prophase, as well as in young spermatid stages after the GRC elimination. We found that in prophase spermatocytes the GRC chromatin differs from that of the regular chromosome complement. The GRC is highly enriched in HP1 beta and exhibits high levels of di- and tri-methylated histone H3 at lysine 9 and tri- and di-methylated histone H4 at lysine 20. The GRC does not exhibit neither detectable levels of di- and tri-methylated histone H3 at lysine 4 nor acetylated histone H4 at lysine 5 and 8. The results prove the heterochromatic organization of the GRC in male germline and strongly suggest its transcriptional inactive state during male prophase. Following elimination, in young spermatids the GRC lacks HP1 beta signals but maintains high levels of methylated histone H3 at lysine 9 and methylated histone H4 at lysine 20. The release of HP1 from the GRC with respect to its elimination is discussed.


Associations between parental and child overweight and obesity.
Results of the analysis showed that parents and children overweight/obesity were significantly correlated. The sample includes 318 pairs of mothers and children, and 336 pairs of fathers and children at the age 11.3 +/- 0.4 years in Trogir, Croatia. Child overweight and obesity were defined according to body mass index (BMI) 25 and 30 equivalents (kg/m2). The prevalence of total overweight in girls was 25.6% and among boys was 20.5%. Mother's weight (p = 0.003) and BMI (p = 0.006) were greater in obese than in other groups of children. Overweight/obese children were more often found among overweight/obese mothers (p = 0.009) and fathers (p = 0.039). Correlation between overweight/obese children and their father (odds ratio 3.2, 95% CI 1.5-6.8) was stronger than between overweight/obese children and their mothers (odds ratio 2.2, 95% CI 1.2-3.9). Associations with mothers' and daughters' overweight/obesity were stronger (p = 0.017) than mothers' and sons'(p = 0.12). Correlations between children's BMI and fathers' BMI (r = 0.265, p < 0.0001) and between children's BMI and mothers' BMI (r = 0.173, p = 0.002) were significant. Children whose parents are overweight/obese look for greater attention in future preventive programme.


"Fatigue meniscal tears": a description of the lesion and the results of arthroscopic partial meniscectomy.
The purpose of this study was to describe the clinical outcomes of partial meniscectomy in patients with "fatigue meniscal tear", which presents as a non-traumatic tear with abrupt onset of symptoms of a radial tear in the transition between the middle and posterior thirds of the meniscus. We prospectively followed 71 patients with "fatigue meniscal tear" (41 women and 30 men, mean age of 63 years, SD 6.9 years) recruited among 497 patients with isolated medial meniscal lesions treated between January 2006 and June 2011. Inclusion criteria were spontaneous abrupt onset knee pain, minor or no trauma, no radiographic or MRI osteoarthritis, no bone oedema, pre-operative magnetic resonance image of medial meniscus tear, and arthroscopic evaluation demonstrating radial or vertical flap tear in the body to posterior horn junction of the medial meniscus. We followed all patients for a minimum of two years and reviewed their clinical symptoms, physical exam, functional outcome, and patient satisfaction at last follow-up. The average follow-up was 4.2 years, with a minimum follow-up of two years. Among the 71 patients, there were 59 (83.1 %) good or excellent results and 12 (16.9 %) poor results. These 12 patients demanded further treatment because of persistent pain, with three of the patients developing subchondral bone fracture. All patient complaints and poor outcomes could be identified in the initial six months after surgery. There was no gender difference in the subgroup analysis. Our findings indicate that patients with "fatigue meniscal tear" benefit from arthroscopic partial meniscectomy, with only 16.9 % reporting unfavourable results. IV, Cohort study or case series.


Cascaded multi-dithering theory for coherent beam combining of multiplexed beam elements.
The Cascaded Multi-Dithering theory, which allows coherent beam combining of M-by-N beam elements, is presented in this paper. The theory of Cascaded Multi-Dithering is briefly introduced, and demonstrated experimentally by combining sixteen-beams to verify its feasibility as an active phase control for scaling up the power of fiber lasers.