Arsenite-induced germline apoptosis through a MAPK-dependent, p53-independent pathway in Caenorhabditis elegans.
Many studies have shown that arsenite is a potent inducer of apoptosis both in cells and tissues. However, there is a lack of appropriate in vivo animal models to study the underlying mechanisms of arsenite-induced apoptosis. Caenorhabditis elegans is an excellent model organism for studying many biological processes. We showed previously that C. elegans could be used as an in vivo system to investigate the genotoxic effects of arsenite. In order to elucidate the underlying mechanisms of arsenite-induced apoptosis in vivo, in the present study, we used the mutated alleles of the C. elegans homologue of known mammalian genes that are involved in the regulation of apoptosis. Our results showed that the loss-of-function mutations of p53/ cep-1 and DNA damage response (DDR) genes hus-1, clk-2, and egl-1 exhibited significant increase in germline apoptosis under arsenite exposure, whereas arsenite-induced germline apoptosis was blocked in loss-of-function alleles of extracellular signal-regulated kinase (ERK) (lin-45 (ku51), mek-2 (n1989), and mpk-1 (ku1)), c-Jun N-terminal kinase (JNK) (jkk-1 (km2), mek-1 (ks54), jnk-1 (gk7), mkk-4 (ju91)), and p38 ( nsy-1 (ag3), sek-1 (ag1), and pmk-1 (km25)) MAPK cascades. These results suggest that arsenite-induced apoptosis occurs independently of p53/ cep-1 and the DNA damage response (DDR) genes hus-1, clk-2, and egl-1 and that the C. elegans caspase gene ced-3, Apaf-1 homologue ced-4, and the MAPK signaling pathways are essential for germline apoptosis. Moreover, our study demonstrates that C. elegans could be a mammalian in vivo substitute model to study the mechanisms of apoptosis.


A comparative study on the cost of new antibiotics and drugs of other therapeutic categories.
Drug treatment is becoming more expensive due to the increased cost for the introduction of new drugs, and there seems to be an uneven distribution of medication cost for different therapeutic categories. We hypothesized that the cost of new antimicrobial agents may differ from that of other therapeutic categories and this may play a role in the stagnation of development of new antibiotics. We performed a pharmaco-economical comparative analysis of the drug cost of treatment for new agents introduced in the United States drug market in various therapeutic categories. We calculated the drug cost (in US dollars) of a ten-day treatment of all new drugs approved by the FDA during the period between January 1997 and July 2003, according to the 2004 Red Book Pharmacy's Fundamental Reference. New anti-neoplastic agents were found to be the most expensive drugs in comparison to all other therapeutic categories, with a median ten-day drug-treatment cost of US$848 compared to the median ten-day drug-treatment costs of all other categories ranging from US$29 to US$301. On the other hand, new antimicrobial drugs were found to be much less expensive, with a median ten-day drug-treatment cost of US$137 and $US85 for all anti-microbial agents and for anti-microbial agents excluding anti-HIV medications, respectively. The drug-treatment cost of new medications varies considerably by different therapeutic categories. This fact may influence industry decisions regarding the development of new drugs and may play a role in the shortage of new antimicrobial agents in the fight against the serious problem of antimicrobial resistance.


Interaction of insulin receptors with lipid bilayers and specific and nonspecific binding of insulin to supported membranes.
In the first part, we study the interaction of the insulin receptor with model membranes of dimyristoylphosphatidylcholine (DMPC) by various techniques, including calorimetry, densitometry, static light scattering, and electron microscopy. By analyzing the pronounced depression of the lipid chain melting transition in terms of the Van Laar-Hildebrand theory of regular dilute solutions, an (exothermic) interaction energy of Wp = 2000 kJ.mol-1 is found for the receptor and of WL = 0.6 kJ.mol-1 for the lipid. This is interpreted in terms of an adsorption of the 2 hydrophilic head groups of the receptor to the membrane surface so that 1 protein interacts with about 2000 lipids. This number is verified by freeze-fracture electron microscopy. Binding of insulin induces a remarkable decoupling of the receptor head group from the membrane, pointing to a pronounced conformational change. In the second part, we introduce a simple fluorescence technique by which adsorption isotherms of water-soluble and fluorescent-labeled substrates, such as insulin, to membranes may be determined. It is based on the selective evanescent field excitation of ligands adsorbed to supported planar bilayers on argon-sputtered glass plates. These are deposited by the monolayer transfer technique or by vesicle condensation. The reconstituted receptor exhibits a weak (binding constant Kw = 3 X 10(9) L.M-1) and a strong (binding constant Ks greater than 10(10) L.M-1) binding site. Insulin exhibits a weak but remarkable nonspecific binding to bilayers of pure DMPC and DMPC containing 20% positively charged lipid and a strong binding to DMPC containing negatively charged lipids such as phosphatidylserine or ganglioside (GT1b).(ABSTRACT TRUNCATED AT 250 WORDS)


Ocular findings among patients with late-diagnosed or poorly controlled homocystinuria compared with a screened, well-controlled population.
Untreated homocystinuria (HCU) leads to systemic and ocular complications preventable by early treatment. This study describes the ocular features in HCU patients who had late diagnosis or were noncompliant with treatment compared with a control group of early-diagnosed and well-controlled subjects. Fourteen late-diagnosed HCU patients with a median age at diagnosis of 4 years (range, 1.25-28 years) were studied. Five patients were born outside of Ireland or before screening began. All 14 patients had lens subluxation or dislocation at diagnosis. Only 28.6% of eyes had 20/40 vision or better. Three patients were tested for HCU following the diagnosis in a sibling. Four patients attended ophthalmology departments for a median of 12.8 years (range, 4-23 years) prior to diagnosis of HCU; all had steadily progressive myopic astigmatism and lens subluxation. Six patients who became poorly controlled in their teens or early twenties showed significant progression of their myopia, and 3 had phacodonesis or lens subluxation develop. All eyes in this group had 20/40 vision or better. Fifteen patients who were detected in the newborn period and remained well controlled had no evidence of lens subluxation. All of the control group patients had 20/20 vision bilaterally. The difference in visual acuity between late-diagnosed patients and the control group was highly significant (P =.0002). The differences in refractive errors between the groups were also highly significant (P =.0001). Lens subluxation is a principal feature of untreated HCU, yet we found a median lag period of 5.5 years in 4 cases before diagnosis. Young persons with marked and progressive myopia or idiopathic lens subluxation should be screened for HCU.


Nucleic acid-phospholipid recognition: Fourier transform infrared spectrometric characterization of ternary phospholipid-inorganic cation-DNA complex and its relevance to chemicopharmaceutical design of nanometric liposome based gene delivery formulations.
The present work is a continuation of our previous microscopic, spectroscopic and microcalorimetric measurements of liposomes and poly(ribo)nucleotides and their ternary complexes with inorganic cations as an alternative formulation employing zwitterionic phospholipids instead of cytotoxic cationic lipids. Current report describes Fourier transform infrared spectrometric study as employed to follow structural transitions of newly proposed ternary solid neutral lipid-Mg(2+)-DNA complexes as promising gene delivery formulation. Spectra of the unbound components are compared with those obtained after their complexation as binary and ternary mixtures. Results are described at the levels of carbonyl, phosphate, choline and CH groups and discussed as effects of nucleic acid and phosphatidylcholine moiety on each other in the absence and in the presence of Mg(2+). The infrared spectra of DNA-lipid phases are dominated by the lipid specific absorption bands, with a very little contribution of DNA. Data suggest that upon recognition of DNA with lipids, the DNA undergoes helical transition. Mg(2+) effects are interpreted as dehydrations of phosphates and H-bonding inducing effects on carbonyl groups. The role of residual and surface water on these associations, as well as on chain packing is also discussed followed by possible implications of the ternary complex formation for further gene transfer designs.


Myocardial Extracellular Volume Fraction Measured by Cardiac Magnetic Resonance Imaging Negatively Correlates With Cardiomyocyte Breadth in a Healthy Porcine Model.
The extracellular volume fraction (ECV) derived from cardiac magnetic resonance imaging (MRI) is extensively used to evaluate myocardial fibrosis. However, due to the limited histological verification in healthy individuals, it remains unclear whether the size of cardiomyocytes may play a potential role in the physiological changes of ECV. The aim of this study was to examine the association between cardiomyocyte size and myocardial ECV by using a healthy porcine model. Sixteen domestic healthy pigs were anesthetized and underwent cardiac MRI with mechanical controlled breathing. Intravenous contrast medium was introduced at a dose of 0.2-0.25 mmol/kg. The interventricular septum ECV was calculated using an established MRI procedure, which was based on the pre- and post-contrast T1 values of the heart and individual blood hematocrit. The cardiomyocyte breadth (CmyB) in cross section was measured by hematoxylin and eosin staining to reflect the cardiomyocyte size. Data were successfully acquired from 14 pigs. The CmyB was obtained from the myocardial tissues corresponding to the region of interest on cardiac MRI. The mean ± SD of the ECV was 0.253 ± 0.043, and the mean ± SD of the CmyB was 10.02 ± 0.84 μm. The ECV exhibited a negative correlation with the CmyB (r = -0.729, p = 0.003). The myocardial ECV detected by cardiac MRI is negatively correlated with the CmyB in healthy pigs, demonstrating that the size of cardiomyocytes is potentially associated with the ECV under physiological conditions.


Double-labelling data on somatostatin-like and tyrosine-hydroxylase-like immunoreactivities in chick embryo brain stem. I. Pons and mesencephalon.
With the aim of investigating some factors and mechanisms of the chicken brain development, the same thick sections of brain stems from twelve E13-to-E21-aged chick embryos were sequentially tested with a rabbit anti-Somatostatin antiserum, using a PAP-DAB technique, and with anti-tyrosine hydroxylase (-TH) monoclonal antibodies, using an indirect immuno-fluorescence technique. As regards the pons and mesencephalon, the following main results were obtained. At E21 almost the same distribution of the TH-like immunoreactivity (ir) as at E13 was observed. Neuroblasts in a central, relatively wide region of mesencephalic tegmentum and in the central portion of the pons showed TH-like ir. A co-localization of the 2 immunoreactivities was detected only at E18, within some neuroblasts of the mesencephalic and pontine regions with TH-like ir. It is possible that this transitory co-localization plays a role in the development of the pons and mesencephalon of this species.


[The theoretical basis and acupoint selection of acupuncture treatment for "chemo brain"].
Through clinical practice and literature analysis, the effectiveness and feasibility of acupuncture and moxibustion for "chemo brain" (chemotherapy-induced cognitive impairment, CICI) are explored, which provides a theoretical basis of acupuncture and moxibustion for "chemotherapy brain". From the point view of TCM, CICI is classified as amnesia-like disease. The location of disease is in brain and it is most closely related to heart and kidney. The pathogenesis is the deficiency of kidney and marrow, often accompanied with stagnation of qi due to depression of liver. It is deficient in root and excessive in superficial. From the point view of modern research, CICI is a cognitive dysfunction disease, which is closely related to the cerebral cortex and hippocampus. Acupuncture can play a neuroprotective role by regulating various mechanisms to improve cognitive dysfunction. The acupuncture prescriptions should focus on regulating liver and nourishing kidney, accompanied with head acupoints to improve brain and intelligence. The electroacupuncture can be applied. Acupuncture has a rich theoretical basis in the treatment of CICI, but there is a lack of research in this area.


Foreign Relations.
In a large Colombian teaching hospital, a fifty-five-year-old woman complaining of stomach pain is examined by a foreign-exchange medical student from the United States. Speaking in Spanish, the student elicits a medical history that suggests a possible recurrence of gallstones, but nothing further. Upon discussing the patient's case in private with the attending physician, the student is shocked to learn that the patient is suffering from terminal, metastatic gastric cancer but is unaware of her diagnosis. The attending physician explains that the patient's husband instructed the health care providers not to tell the patient what her diagnosis is. The medical student feels uncomfortable withholding the diagnosis from the patient. The thought of disclosure feels equally uncomfortable, however, as it would mean contradicting the instructions of the attending physician at the host hospital as well as violating what the student suspects, but is not sure, may be acceptable medical practice in Colombia. How should the medical student proceed?


[Diagnosis and treatment of hydronephrosis: report of 100 cases].
One hundred patients with hydronephrosis were treated for 10 years. In 88 patients operations confirmed the causes of obstruction including pyelo-ureteral junction obstruction (pujo, 46 cases), anomalous (15), retroperitoneal fibrosis (15), ureteral polyps and valves and others (12). In accordance with etiology and renal function pyeloplasty was carried out in 31 cases, nephrectomy 37 and nephrostomy 12; other operation was done in 20 cases. The successful rate of pyeloplasty was 91.3%. Early diagnosis and treatment of hydronephrosis is key for reserving renal function. Renal r-scan, percutaneous nephrostomy are useful in judging renal function. Dual-Flex stent internal drainage is better than external drainage when pyeloplasty is done.


[Ultrastructural observations on amastigote forms of Leishmania donovani in a case of laryngeal leishmaniasis].
An ultrastructural study on the amastigote stage of Leishmania donovani, in a rare case of laringeal involvement, was carried out. Many details of the fine morphology of this protozoon were observed under the electron microscope, i.e.: the outer membrane of host origin, periplast, subpellicular microtubules, kinetoplast, flagellum, flagellar pocket, multivesicular bodies. A presence of a new organelle (4 microtubules structure close to and running parallel to the flagellar pocket), previously not described in Leishmania donovani's species, was demonstrated in this indigenous strain (from Central Italy). The Authors found a limited number of subpellicular microtubules in this strain, i.e.: 72 microtubules. Differences in size and number of microtubules between strains must be investigated in more rigorous morphometric studies. It's possible that the number of the subpellicular microtubules represents a new taxonomic character.


Prevalence of colorectal polyps in a group of subjects at average-risk of colorectal cancer undergoing colonoscopic screening in Tehran, Iran between 2008 and 2013.
Colorectal cancer (CRC) is one of the prime causes of mortality around the globe, with a significantly rising incidence in the Middle East region in recent decades. Since detection of CRC in the early stages is an important issue, and also since to date there are no comprehensive epidemiologic studies depicting the Middle East region with special attention to the average risk group, further investigation is of significant necessity in this regard. Our aim was to investigate the prevalence of preneoplastic and neoplastic lesions of the colon in an average risk population. A total of 1,208 eligible asymptomatic, average- risk adults older than 40 years of age, referred to Firuzgar Hospotal in the years 2008-2012, were enrolled. They underwent colonoscopy screening and all polypoid lesions were removed and examined by an expert gastrointestinal pathologist. The lesions were classified by size, location, numbers and pathologic findings. Size of lesions was measured objectively by endoscopists. The mean age of participants was 56.5±9.59 and 51.6% were male. The overall polyp detection rate was 199/1208 (16.5 %), 26 subjects having non-neoplastic polyps, including hyperplastic lesions, and 173/1208 (14.3%) having neoplastic polyps, of which 26 (2.15%) were advanced neoplasms .The prevalence of colorectal neoplasia was more common among the 50-59 age group. Advanced adenoma was more frequent among the 60-69 age group. The majority of adenomas were detected in the distal colon, but a quarter of advanced adenomas were found in the proximal colon; advance age and male gender was associated with the presence of adenoma. It seems that CRC screening among average-risk population might be recommended in countries such as Iran. However, sigmioidoscopy alone would miss many colorectal adenomas. Furthermore, the 50-59 age group could be considered as an appropriate target population for this purpose in Iran.


Visualization of calcium influx through channels that shape the burst and tonic firing modes of thalamic relay cells.
Thalamic neurons have two firing modes: "tonic" and "burst." During burst mode, both low-threshold (LT) and high-threshold (HT) calcium channels are activated, while in tonic mode, only the HT-type of calcium channel is activated. The calcium signals associated with each firing mode were investigated in rat thalamic slices using whole cell patch clamping and confocal calcium imaging. Action potentials were induced by direct current injection into thalamic relay cells loaded with a fluorescent calcium indicator. In both tonic and burst firing modes, large calcium signals were recorded throughout the soma and proximal dendrites. To map the distribution of the channels mediating these calcium fluxes, LT and HT currents were independently activated using specific voltage-clamp protocols. We focused on the proximal region of the cell (up to 50 microm from the soma) because it appeared to be well clamped. For a voltage pulse of a given size, the largest calcium signals were observed in the proximal dendrites with smaller signals occurring in the soma and nucleus. This was true for both LT and HT signals. Rapid imaging, using one-dimensional linescans, was used to more precisely localize the calcium influx. For both LT and HT channels, calcium influx occurred simultaneously throughout all imaged regions including the soma and proximal dendrites. The presence of sizable calcium signals in the dendrites, soma, and nucleus during both firing modes, and the presence of LT calcium channels in the proximal dendrite where sensory afferents synapse, have implications for both the electrical functioning of relay cells and the transmission of sensory information to cortex.


Effects of age and caloric restriction on brain neuronal cell death/survival.
Aging may pose a challenge to the central nervous system, increasing its susceptibility to apoptotic events. Recent findings indicate that caloric restriction (CR) may have a profound effect on brain function and vulnerability to injury and diseases, by enhancing neuroprotection, stimulating the production of new neurons, and increasing synaptic plasticity. Apoptosis and apoptotic regulatory proteins in the brain frontal cortex of 6-month-old ad libitum fed (6AD), 26-month-old ad libitum fed (26AD), and 26-month-old caloric-restricted (26CR) male Fischer 344 rats (40% restriction compared to ad libitum fed) were investigated. Levels of Poly-ADP ribose polymerase (PARP-DNA repair enzyme; its cleaved 89 kDA fragment is a marker of apoptosis), cytoplasmic histone-associated DNA fragments, and X chromosome-linked inhibitor of apoptosis (XIAP--an endogenous apoptosis inhibitor) were determined. A significant age-associated increase in PARP was found, which was ameliorated in the frontal cortices of the CR rats. No significant differences in cytoplasmic histone-associated DNA fragments with age or with CR were observed. XIAP levels significantly increased with age in the brains of the ad libitum animals, while CR animals exhibited the highest levels of this inhibitor compared to all groups. Our findings suggest that caloric restriction may provide neuroprotection to the aging brain by preserving DNA repair enzymes in their intact form, and/or upregulating specific antiapoptotic proteins involved in neuronal cell death.


Evidence for high inter-generational individual quality in yellow-eyed penguins.
Longitudinal studies focusing on lifetime reproductive success (LRS) have been used to measure individual breeding performance and identify commonalities among successful breeders. By extending the focus to subsequent generations we identify a proportion of high-quality individuals that contribute disproportionately to the population over multiple generations. We used 23 years of yellow-eyed penguin (Megadyptes antipodes) breeding data from one breeding area to identify the proportion of individual birds that raised successful breeders, which in turn raised offspring. We explored which life-history components influenced LRS, as this knowledge would enable conservation resources to be focused on high-performing individuals in this endangered population. From 2,147 birds marked as chicks, 370 (17.2%) survived to adulthood and recruited to their natal location, of which 219 (10.2%) fledged offspring: 124 (56.6%) of the 219 birds produced offspring that recruited as breeders. Only 102 birds (4.8% of 2,147) fledged first-generation offspring that in turn fledged offspring (second-generation offspring, or grand-offspring). We found that ∼25% of the birds that survived to breed had above-average LRS as well as above-average numbers of grand-offspring, and were more likely to have produced first-generation chicks that recruited and also produced above-average numbers of second-generation chicks. Our findings suggest that there is a core of "super-breeders" that contribute disproportionately to the population over successive generations. Lifespan and age-at-first-breeding were correlated with LRS. We suggest that traits of birds relating to longevity, health (e.g., immunocompetence) and fitness could be examined to identify potential links with high LRS and inter-generational fecundity. "Super-breeders" appear to consistently achieve high LRS and long lifespans in a stochastic environment, demonstrating greater resilience in the face of extreme events.


Effect of oxLDL on transcriptional expression of human lens epithelial cells.
Age-related cataract patients regularly have hypertension, hyperglycemia, and hyperlipidemia. In oxidative conditions, increased reactive oxygen species can oxidize natural low-density lipoprotein into oxidative low-density lipoprotein (oxLDL). However, the relationship between oxLDL and the occurrence of cataracts is still unclear. In this study, 1515 differentially expressed transcripts were identified by analyzing the results of RNA sequencing in a human lens epithelial cell line (HLEpiC). Compared with control groups, oxLDL-treated HLEpiC had 806 up-regulated transcripts and 709 down-regulated transcripts. Our genome-wide transcriptome results showed that differentially expressed genes, such as Rho signaling (Rho A and Cdc42) and Na+/K+-ATPase family (ATP1B1), are involved in lens epithelial cell differentiation and cell homeostasis. In conclusion, oxLDL greatly influences transcriptional expression and these differentially expressed genes may play an important role in the development of cataracts. Our findings may provide new targets in the treatment for cataracts.


Assessing unit-price related remifentanil choice in rhesus monkeys.
Given a commodity available at different prices, a unit-price account of choice predicts preference for the cheaper alternative. This experiment determined if rhesus monkeys preferred remifentanil (an ultra-short-acting micro-opioid agonist) delivered at a lower unit price over a higher-priced remifentanil alternative (Phases 1 and 3). Choice between equal-priced alternatives also was assessed (Phase 2). A discrete-trials procedure was arranged in which three monkeys chose between two remifentanil alternatives by responding on one of two levers. Different prices were arranged by manipulating drug dose (0.3 and 0.1 microg/kg/injection) and/or the ratio requirement. Monkeys usually chose the larger-dose alternative even when it was more expensive. Only when unit prices were relatively high (e.g., large response requirements) did monkeys choose the cheaper (or equally priced) smaller-dose alternative. Employing larger doses (0.9 and 0.3 microg/kg/injection) attenuated the larger-dose preference. The results demonstrate that choice was not determined simply by unit price. An alternative model that employs demand-function analysis to generate choice predictions is proposed.


The effect of an oral ginger supplementation on NF-κB concentration in peripheral blood mononuclear cells and anthropomorphic data of patients with type 2 diabetes: A randomized double-blind, placebo-controlled clinical trial.
The complications of diabetes are extensive which can be caused by excessive oxidative stress, inflammation and impaired insulin system. Plant-sourced bioactive compounds can reduce inflammation and oxidative stress. The aim of present study was to determine the effect of ginger supplementation on diabetic complications. The present study is a randomized double blind clinical trial which is conducted with 48 diabetic patients. The participants were randomly divided into two intervention and placebo groups which were received 2 g ginger powder and 2 g wheat flour respectively for 10 weeks. Nuclear factor kappa B (NF-κB) concentration and anthropometric measurements were evaluated at the baseline and at the end of study. The effect of ginger supplementation on hip circumference was marginal and there was no significant effect on BMI and waist circumference. Mean NF-κB p65 concentrations were reduced in ginger supplementation group, however, the amount was not statistically significant. Ginger supplementation had significant effects on anthropometric evaluations. Ginger supplementation decreased mean NF-κB concentration in comparison with placebo, however the significance level was marginal. In order to achieve reliable information, more researches should be complemented with uptake of other diagnostic tools.


Intrapericardial cystic hematoma in a dog treated by thoracoscopic subtotal pericardectomy.
CASE DESCRIPTION A 2-year-old castrated male mixed-breed dog was evaluated because of a 1-week history of respiratory distress and abdominal distension. Thoracic radiography and echocardiography at that time revealed an enlarged cardiac silhouette and pericardial effusion; abdominal radiography and ultrasonography revealed ascites. CLINICAL FINDINGS At the initial referral examination 5 weeks later, the dog weighed 37.5 kg (82.5 lb) and appeared clinically normal. The only abnormality detected was a grade I/VI systolic murmur on the left side of the thorax. Echocardiography revealed a large fat- and fluid-filled cystic structure located next to the right ventricle with scant pericardial effusion. Computed tomography revealed a bilobed peripherally contrast-enhancing structure within the right ventral aspect of the pericardium; the right ventricle appeared compressed by the cyst. TREATMENT AND OUTCOME Initial treatment consisted of pericardiocentesis and abdominocentesis to alleviate clinical signs. Thoracoscopic subtotal pericardectomy was performed 6 weeks after the initial treatment. The cyst was completely excised, and multiple adhesions between the visceral and parietal pericardium were transected, without surgical or anesthetic complications. Histologic examination of the cyst revealed chronic inflammation with histiocytic infiltration, suggesting possible foreign body reaction or chronic inflammation and hemorrhage. These findings supported a diagnosis of cystic hematoma of the pericardium. The dog remained clinically normal for at least 16 months after surgery. CLINICAL RELEVANCE This report represents a rare case of intrapericardial cystic hematoma in a dog. Minimally invasive surgery was performed without complications, suggesting that thoracoscopic subtotal pericardectomy is a feasible treatment option for affected dogs.


Involvement of platelet-activating factor in ultraviolet B-induced hyperalgesia.
Ultraviolet B (UVB) radiation causes cutaneous inflammation. One important clinical consequence of UVB-induced inflammation is increased pain or hyperalgesia, which is likely mediated by enhanced sensitivity of cutaneous sensory neurons. Previous studies have demonstrated that UVB radiation generates the lipid mediator, platelet-activating factor (PAF), as well as oxidized phospholipids that act as PAF-mimetics. These substances exert effects through the PAF receptor (PAF-R). This study was designed to assess whether PAF-R is involved in UVB-induced hyperalgesia. Intradermal injection of carbamoyl PAF (CPAF; 1-hexadecyl-2-N-methylcarbamoyl glycerophosphocholine) resulted in an enhanced response to mechanical stimuli in wild-type mice but not in PAF-R knockout (KO) mice. There was no significant change in paw withdrawal to noxious thermal stimuli in either genotype after intradermal injection of CPAF. Exposure of the hind paw to 1,500 J m(-2) UVB radiation caused an increased sensitivity to both mechanical and thermal stimulation in wild-type mice but not in PAF-R KO mice. The thermal hyperalgesia caused by UVB irradiation was inhibited in mice that lacked PAF-R in bone marrow-derived cells. These data demonstrate that the PAF-R is important for UVB-induced hyperalgesia. Further investigation of the role of PAF-R signaling in UVB-induced hyperalgesia could provide better understanding of the pathological processes initiated by UVB-induced skin damage.


A reexamination of poneratoxin from the venom of the bullet ant Paraponera clavata.
In 1991, Piek et al. [45] described a voltage-gated sodium channel (VGSC) modifier from "bullet ant" (Paraponera clavata) venom they called poneratoxin (PoTx). Using UV chromatography and Edman degradation they showed two "identical peptides" of 25 residues. We reinvestigated PoTx using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-TMS). De novo sequencing showed the two peptides were actually structurally different peptides: the originally described PoTx and a glycyl pro-peptide (glycyl-PoTx) that lacks C-terminus amidation. We examined P. clavata venom from different geographical locations and discovered two additional PoTx analogs: an A23E substitution analog and a D22N; A23V substitutions analog. We tested PoTx and these three natural analogs on the mammalian sensory voltage-gated sodium channel, Nav1.7, using whole cell voltage-clamp. PoTx and each analog induced slowly activating currents in response to small depolarizing steps and sustained currents due to blockade of channel inactivation, similar to that described previously in skeletal muscle [19]. Glycyl-PoTx had the same potency and efficacy as PoTx. A23E PoTx, with a decrease in both C-terminal net positive charge and hydrophobicity, had an eight-fold reduction in potency compared to PoTx. In contrast, the D22N; A23V PoTx, with an increase in both C-terminal net positive charge and hydrophobicity, had a nearly five-fold increase in potency compared to PoTx. We found that changes in PoTx C-terminus caused a significant change in PoTx potency.


[The relative biological effectiveness (RBE) of fast neutrons on the Dunning rat prostate carcinoma R3327-HI].
Human prostate tumors are known to be good candidates for neutron therapy. The Dunning rat prostate tumor system R3327 was found in many studies to be an excellent model for human prostate tumors. There is still a paucity of studies on the response of the Dunning tumors to fast neutrons. Tumors of the R3327-HI subline are moderately well differentiated and mucin producing. They show one euploid cell population, a bromodeoxyuridine labelling index of 5%, a potential doubling time of 8.9 days, a volume doubling time of about ten days and a cell loss rate of 10%. Tumors were transplanted s.c. in the distal thigh of Copenhagen rats and treated with 60Co-photons (10, 20, 30, 40 Gy, 45 cGy/min) and 14-MeV-neutrons (8, 10, 12 Gy, 7 to 11 cGy/min). Tumor volumes were measured twice weekly. Growth delay was defined as time in days until the tumors reached twice their treatment volume. Linear regressions on the median growth delays of the different treatment groups were calculated. The ratio of the neutron- and photon-slopes yielded an RBE of 3.1 +/- 0.3. Additionally isoeffect-RBE values between 2.3 and 2.6 were graphically estimated.


Separation and preconcentration of Ag(I) in aqueous samples by flotation as an ion-associate using iodide and ferroin followed the determination by flame atomic absorption spectrometry.
A simple method for separation/preconcentration and determination of Ag(I) in aqueous samples is described. The method is based on formation of an ion-associate between Ag(I)-iodide complex and ferroin, which can be floated at the interface of the aqueous/n-heptane phases. The flotation process was carried out using 500-ml aliquot of the aqueous solution and the floated layer was dissolved in 5 ml of 1 M HNO3 containing methanol (50% v/v) as the solvent. The Ag(I) content was then determined by flame atomic absorption spectrometry (FAAS). The method so could be considered as an enrichment process, was achieved to a quantitative feature, when the pH of the solution was adjusted to 4 and the concentrations of iodide and ferroin were about 3.2 x 10(-4) M and 6.25 x 10(-5) M, respectively. The LOD and RSD (n = 7) were obtained 1.0 x 10(-8) M and 2.4%, respectively. It was found that a large number of cations and anions even at high considerably foreign ion/Ag(I) ratios were not interfered. The method was applied satisfactorily to recovery of Ag(I) from different aqueous samples.


Esophagogastric metaplasia relates to nodal metastases in adenocarcinoma of esophagus and cardia.
Immunohistochemical profiles of esophageal and cardia adenocarcinoma differ according to the presence or absence of Barrett's epithelium (BIM) and gastric intestinal metaplasia (GIM) in the fundus and antrum. Different lymphatic spreading has been demonstrated in esophageal adenocarcinoma. We investigated the correlation among the presence or absence of intestinal metaplasia in the esophagus and stomach and lymphatic metastases in patients who underwent radical surgery for esophageal and cardia adenocarcinoma. The mucosa surrounding the adenocarcinoma and the gastric mucosa were analyzed. The BIM+ patients underwent subtotal esophagectomy and gastric pull up, and the BIM- patients underwent esophagectomy at the azygos vein, total gastrectomy, and esophagojejunostomy. The radical thoracic (station numbers 2, 3, 4R, 7, 8, and 9) and abdominal (station numbers 15 through 20) lymphadenectomy was identical in both procedures except for the greater curvature. One hundred ninety-four consecutive patients were collected in three major groups: BIM+/GIM-, 52 patients (26.8%); BIM-/GIM-, 90 patients (46.4%); BIM-/GIM+, 50 patients (25.8%). Two patients (1%) were BIM+/GIM+. A total of 6,010 lymph nodes were resected: 1,515 were recovered in BIM+, 1,587 in BIM-/GIM+, and 2,908 in BIM-/GIM- patients. The percentage of patients with pN+ stations 8 and 9 was higher in BIM+ (p=0.001), and the percentage of patients with pN+ perigastric stations was higher in BIM- (p=0.001). The BIM-/GIM- patients had a number of abdominal metastatic lymph nodes higher than did the BIM-/GIM+ patients (p=0.0001). According to the presence or absence of BIM and GIM in the esophagus and cardia, adenocarcinoma correspond to three different patterns of lymphatic metastasization, which may reflect different biologic and carcinogenetic pathways.


Intracellular pathway and kinetics of protein secretion in the coagulating gland of the mouse.
The coagulating gland of male rodents is part of the prostatic complex. Various mechanisms of secretion have been postulated, in part because organelles commonly involved in the secretory process possess unusual features, such as extreme distension of the rough endoplasmic reticulum. In the present study, the pathway, kinetics, and mode of secretion in the coagulating gland of the mouse were studied by electron microscope autoradiography at intervals between 5 min and 8 h after administration of 3H-threonine. The percentage of grains associated with the rough endoplasmic reticulum was initially high and generally decreased throughout the experiment, while a pronounced rise in the proportion of grains associated with the Golgi apparatus and secretory granules was observed 6 h after injection of precursor. In addition, there was a smaller elevation in the percentage of grains over the Golgi apparatus and secretory granules between 1 and 4 h, and radioactive material first reached the lumen of the gland 4 h after injection of the precursor. Although the general pathway of intracellular transport of secretory protein resembles that in other cells, the results indicate that there are several unusual aspects to the secretory process in the coagulating gland. First, the rate of transport was markedly slower than in most other exocrine gland cells, since the bulk of the labeled protein did not reach the Golgi apparatus and secretory granules until 6 h after administration of precursor. This reflected prolonged retention of secretory products in the endoplasmic reticulum. Second, in addition to the major bolus of labeled material that traversed the cells at about 6 h, a smaller wave of radioactivity appeared to pass through the Golgi apparatus and secretory granules and reach the lumen earlier, within the first few hours after the injection. Finally, the primary mode of secretion in the coagulating gland appears to be merocrine because the secretory granules contained much labeled protein.


[Main causes of poor efficacy of outpatient and hospital medical help to patients with septic forms of the diabetic foot syndrome].
The treatment results of 2431 patients with the septic forms of the diabetic foot syndrome were analyzed. The first group of patients (n=1356) were treated in specialized hospital departments, where as the second group (n=1066) received help in general "septic" departments of city hospitals. Certain organizational, diagnostic and treatment mistakes were registered predominately among patients of the second group. The obtained results vividly indicate the economic efficacy of the development and creation of the specialized departments or centers for the adequate treatment of such patients. It would allow the decrease of amputation rates from 36,9 to 12,0% and lethality rate from 9,8 to 4,3% in specialized departments and general hospitals, relatively.


Recurrent laryngeal nerve injury and hypoparathyroidism rates in reoperative thyroid surgery.
Reoperative thyroid surgery is rare and has a high complication rate. This retrospective cohort study was performed to determine the recurrent laryngeal nerve injury and hypoparathyroidism rates after reoperative thyroid surgery in two university hospitals in Benghazi, Libya. All consecutive patients who underwent reoperative thyroid surgery between January 2002 and July 2014 were included retrospectively. The cohort was divided according to whether the reoperation was in the previously operated lobe or both lobes (ipsilateral group), or only in the previously non-operated lobe (contralateral group). Of the 73 patients, 66 were female and seven were male. The median age was 37 (19-80) years. Nine (12.3%), five (6.8%), and one (1.4%) patient developed postoperative transient hypocalcemia, transient recurrent laryngeal nerve palsy, and permanent recurrent laryngeal nerve injury, respectively. None of the patients developed permanent hypocalcemia. The ipsilateral group had a higher rate of permanent recurrent laryngeal nerve injury after reoperation than the contralateral group (3.1% vs. 0%). It also had higher rates of transient recurrent laryngeal nerve injury (12.5% vs. 2.4%) and transient hypocalcemia (28.1% vs. 0%), but the two groups did not differ in terms of permanent hypocalcemia rates (both 0%). Reoperative thyroid surgery is technically challenging with a high incidence of complications. The ipsilateral group had more complications after reoperative thyroid surgery than the contralateral group. Hemi- or total thyroidectomy at the primary surgery is recommended to reduce the frequency of reoperative thyroid surgery.


Mental health and quality of life of elderly people in the Bethlehem district: a cross-sectional study.
The focus of this study was quality of life and mental health in elderly people living in the Bethlehem district. The aim of this study was to identify the most prevalent mental health problems and how they affect quality of life. This cross-sectional study was done in June and July, 2010. We recruited a convenience sample of elderly people (aged 65 years and older) living in the Bethlehem district of occupied Palestinian territory. Using a structured questionnaire, we assessed quality of life with the WHOQOL-BREF and current psychological status and distress with the Brief Symptom Inventor. Verbal consent form was obtained from all participants before their interview. 291 (97%) of 300 elderly people approached for this study completed the questionnaire. 101 (35%) participants described severe or very severe levels of somatisation. 39 (14%) participants reported severe or very severe obsessive compulsive symptoms, 27 (9%) complained of severe or very severe anxiety symptoms, and 37 (13%) had severe or very severe depression. 135 (46%) participants rated their quality of life as good or very good, but the physical health domain of the WHOQOL-BREF scale had the lowest scores (mean 50·9 [SD 21·4]) of all the domains. 103 (36%) participants perceived their physical health as being good or very good, whereas 95 (33%) participants rated their social life as good or very good. 117 (40%) participants were satisfied or very satisfied with their environment, and 119 (41%) participants evaluated their psychological health as good or very good. A stepwise regression analysis of variables indicated that an increase in psychological symptoms was independently associated with lack of education, living in a rural area, being economically dependent on others, and living with chronic disease. Poor quality of life was independently associated with increased age, lack of education, living in the home of sons, and being economically dependent on others. In view of the high prevalence of chronic diseases and their negative effect on psychological health and quality of life, we recommend the provision of better health care for elderly people, especially those living in rural areas. Our findings also show the importance of special education programmes for the physical and psychological health of elderly people. None.


Effects of mutations in the hinge region of serpins.
An expression system for alpha 1-antitrypsin in Escherichia coli was developed using a T7 RNA polymerase promoter. Addition of rifampicin to inhibit the E. coli RNA polymerase after induction of the T7 RNA polymerase gene resulted in about 30% of newly synthesized protein being alpha 1-antitrypsin. This expression system was then used to examine the effect of mutations in the hinge region of alpha 1-antitrypsin on its activity. The mutations were based on ones in antithrombin III that had previously been shown to have adverse effects on activity. Mutation of Ala347 to threonine in alpha 1-antitrypsin did not affect the kinetic behavior of the protein with trypsin or human leukocyte elastase. In contrast, mutation of Gly349 to proline converted the majority of the protein into a substrate for both proteinases. The small fraction of this mutant that was active, however, had kinetic parameters that were indistinguishable from wild-type alpha 1-antitrypsin. Cleavage within the reactive-site loop of wild-type alpha 1-antitrypsin causes a conformational change in the molecules (the S-to-R transition) and results in a marked increase in heat stability. This increase in heat stability was also seen upon cleavage within the reactive-site loops of both of the alpha 1-antitrypsin mutants. The results are discussed in terms of a kinetic mechanism for serpin-proteinase interactions, in which after the formation of an initial complex the serpin partitions between the formation of a stable complex and a cleavage reaction.(ABSTRACT TRUNCATED AT 250 WORDS)


[Congenital haemangiomas and other rare infantile vascular tumours].
A number of infantile tumours, far less frequent than infantile haemangiomas, were long assimilated to them. Today they are clearly individualised, based on distinctive clinical and pathologic features, and this difference has been supported by the discovery of new immunophenotypic markers such as GLUT1. GLUT1 stains 100% of infantile haemangiomas and none of the other infantile vascular tumours. Congenital haemangiomas represent a group of vascular tumours still under evaluation as they have slightly heterogeneous presentation. Their prognosis is better appraised and their therapeutic management has improved. They are all fully grown in utero and they do not experience postnatal proliferation like haemangiomas do. Some of them (RICH--Rapidly Involuting Congenital Haemangioma) undergo spontaneous involution during the first year. Others (NICH--Non Involuting Congenital Haemangioma) persist lifelong. Tufted angioma and kaposiform haemangioendothelioma are histopathologically well characterized; in addition they are now considered as part of a same spectrum of vascular tumours, with the contribution of lymphatic endothelial cells in their proliferation. Both are clearly the tumours able to create platelet trapping, thrombocytopenia and the life-threatening Kasabach-Merritt syndrome. However they may occur as isolated tumours, without thrombocytopenia but with cosmetic, and sometimes function-impairing, consequences.


Single-fiber EMG in familial hemiplegic migraine.
Twelve familial hemiplegic migraine (FHM) patients (6 with the I1811L mutation in CACNA1A, 3 with M731T mutation in ATP1A2, and 3 without known mutations) and 10 control subjects underwent single-fiber EMG. Mean jitter did not differ significantly between patients and control subjects or among patients. No blocking was found. The results suggest that neuromuscular function is normal in FHM.


Home Urodynamic Pressures and Volume Measurement for the Neurogenic Bladder: Initial Validation Study.
We evaluated the ability of a bladder pressure/volume diary to identify patients at risk for increased intravesical pressures. Patients dependent on clean intermittent catheterization used ruler based manometry to measure intravesical pressures before leakage or scheduled drainage at home. We prospectively collected clinical, urodynamic and bladder pressure/volume diary data in patients with spina bifida who were optimized on anticholinergic therapy and clean intermittent catheterization. Measurements were taken with patients in the supine position with relaxed abdominal muscles. We defined increased pressure as detrusor pressure greater than 30 cm water as measured by urodynamics. ROCs were plotted to correlate bladder pressure/volume diary variables with abnormal intravesical pressures, and the most sensitive variable in determining abnormal intravesical pressures was sought as the end point. A total of 30 patients with a mean age of 10 years (range 1 to 20) were included. Home pressures measured at maximal clean intermittent catheterization volume and mean bladder pressure/volume diary pressures were most reliable in predicting urodynamic pressures greater than 30 cm water (AUC 0.93 and 0.87, respectively). Home pressures measured at maximal clean intermittent catheterization volumes less than 20 cm water were associated with normal bladder pressures (less than 30 cm water) on urodynamics, with a sensitivity of 100% and a specificity of 80%. Home manometry less than 20 cm water provides a reliable measurement of safe pressures. A bladder pressure/volume diary is feasible and can aid in monitoring pressures at home without the additional cost and morbidity of urodynamics. A bladder pressure/volume diary may be a useful tool to help identify patients who would benefit from urodynamic testing.


Peer review comments on drug trials submitted to medical journals differ depending on sponsorship, results and acceptance: a retrospective cohort study.
During peer review, submitted manuscripts are scrutinised by independent experts to assist journal editors in their decision-making and to help improve the quality of articles. In this retrospective cohort study, peer review comments for drug trials submitted to medical journals were analysed to investigate whether there is a relation between the content of these comments and sponsorship, direction of results and decision about acceptance. Descriptive content analysis of reviewer comments made on manuscripts on drug trials submitted to eight medical journals (January 2010-April 2012). For each manuscript, the number of reviewers, decision about acceptance, sponsorship and direction of results were extracted. Reviewer comments were classified using a predefined checklist. Reviewer reports for 246 manuscripts were assessed. Industry-sponsored trials were more likely to receive comments about lack of novelty (8.9%) than industry-supported (2.5%) and non-industry trials (6.1%, overall p=0.038). Non-industry trials more often received comments about poor experimental design (69.7%) than industry-supported (58.8%) and industry-sponsored trials (52.9%, overall p=0.019). Non-industry trials were also more likely to receive comments regarding inappropriate statistical analyses (28.4%) than industry-supported (23.5%) and industry-sponsored trials (15.1%, overall p=0.006). Manuscripts with negative results were more likely to receive comments about inappropriate conclusions (29.3%) than those with positive results (18.9%, p=0.010). Rejected manuscripts had more often received comments on the research question not being clinically relevant (7.8%) than accepted manuscripts (1.6%, p=0.002), and also on lack of novelty (8.3% vs 2.6%, p=0.008) and poor experimental design (68.6% vs 50.5%, p<0.001). Reviewers identified fewer shortcomings regarding design and statistical analyses in industry-related trials, but commented more often on a lack of novelty in industry-sponsored trials. Negative trial results did not significantly influence the nature of comments other than appropriateness of the conclusion. Manuscript acceptance was primarily related to the research question and methodological robustness of studies.


Translucency and masking ability of various opaque-shade composite resins.
The aims of this study were to evaluate the adequate thickness of opaque resins for situations such as an oral black cavity and discolored tooth structure, as well as the translucency of each opaque material at various thicknesses. Six opaque-shade composite resins (Z-350 OA3, Amelogen Universal A2O, Esthet-X A2O, Esthet-X A4O, Charmfil UO, and Aelite Universal OA3) were prepared in metal molds with a hole of 8mm in diameter and various thicknesses (0.5mm, 1.0mm, 1.5mm, 2.0mm, 2.5mm, 3.0mm and 4.0mm). Four backgrounds (white tile, black tile, C4 shade porcelain and opaque resin itself) were used to determine the translucency parameter (TP) (between black and white backgrounds), and to mimic a black oral cavity (between black and opaque resin backgrounds) and a discolored tooth structure (between C4 and opaque resin backgrounds). Color measurements were made by a colorimeter to determine the CIELAB values of each specimen with each background and to calculate the TP and DeltaE* value difference among the specimens on the backgrounds. TP and DeltaE* obtained between black and opaque resin backgrounds decreased in similar pattern as thickness increased. A C4 background was masked by resin thicknesses of 0.5-1.0mm, while a black background required thicknesses of 1.0-2.0mm. This study can provide helpful guidelines for the esthetic restoration of discolored tooth structures or 'through and through' class III and IV cavities with composite resin materials.


CD4+CD25+ regulatory T cells protect against injury in an innate murine model of chronic kidney disease.
Studies of mechanisms of disease regulation by CD4+CD25+ regulatory T cells (Treg) have been focused on their interaction with effector T cells; however, the possibility that regulation might involve noncognate cells has not been explored in detail. This study investigated the effect of CD4+CD25+ Treg on macrophage proinflammatory properties and phenotype in vitro and found that they modulate macrophages by inhibiting their activation, leading to reduced proinflammatory cytokine production and a downregulated effector phenotype. For testing the in vivo significance of this effect, CD4+CD25+ T cells that expressed high levels of Foxp3 were reconstituted into SCID mice after induction of Adriamycin nephropathy, a noncognate model of chronic renal disease. CD4+CD25+ T cells significantly reduced glomerular and interstitial injury. In addition, there was a significant fall in the number of macrophages in both the glomeruli and interstitium of SCID mice that were reconstituted with Treg as compared with the Adriamycin alone group. Blockade of TGF-beta using neutralizing antibodies significantly impaired the protective effect of Treg. These findings delineate a TGF-beta-dependent Treg-macrophage inhibitory interaction that can explain cognate-independent protection by Treg.


[The use of deffated corn germ in flour manufactured to bakery products].
The flour of deffated corn germ is a by-product of the corn industry milling dried which is actually subutilized in human consumption. In this study, two samples of corn deffated germ of different particle size were physicochemically characterized: fine fraction (FF) and medium fraction (FM) and they were added to wheat flours to obtain composite flours with substitution levels of 5- 10- 12- 15% mixtures. These mixtures were used to manufacture sandwich bread in bakeries, which were physico-chemically and sensorially analyzed. (FF) and (FM) were found to provide from 20 to 35% of dietary fiber. From the manufactured bread loaves, the ones containing 10% of (FM) and 10% of (FF) were selected because they did not differ in specific weight, color, flavor and texture at a 95% confiability from those elaborated with 100% wheat flour. In the acceptability test given to 200 consumers, it was found that bread with 10% FF substitution was more widely accepted (48%) than that containing 100% wheat flour; and that of 10% of (FM) had the same level of acceptability (28%) as the control bread. It can be concluded that the deffated corn kernel represents a raw material available in this country that could be used in bread elaboration as a partial replacement for imported wheat.


Venous thromboembolism following minimally invasive surgery among women with endometrial cancer.
To determine the rate of venous thromboembolism (VTE) among women undergoing minimally invasive surgery (MIS) for endometrial cancer. Women undergoing robotic or laparoscopic hysterectomy for endometrial carcinoma or complex hyperplasia with atypia were identified between January 2009 and 2014 in a community based health care system. Patient data including age, race, cancer stage, grade, procedure type, length of hospital stay, use of prophylaxis, and diagnosis of VTE were collected retrospectively. The primary outcome was the rate of VTE within 30days following surgery. Fischer's exact tests were performed to evaluate factors associated with VTE. During the study period, 1433 patients underwent MIS for endometrial cancer, with 20 excluded due to known thrombophilia, VTE history, or long-term anticoagulation. A total of 1413 patients were included (739 robotic and 674 laparoscopic cases). All women received mechanical prophylaxis per hospital policy and 61% had additional pharmacologic prophylaxis. The rate of VTE was 0.35% (5/1413), which did not differ among those who received pharmacologic compared to mechanical prophylaxis (0.23% [2/865] versus 0.55% [3/548] respectively, p=0.38). No factors were associated with increased risk of VTE due to the low event rate. VTE in patients undergoing MIS for endometrial cancer was very low irrespective of the mode of prophylaxis received in this large cohort. National guidelines for VTE prophylaxis need to differentiate the low risk associated with MIS surgery from the risk associated with laparotomy for endometrial cancer. We recommend mechanical prophylaxis is sufficient for these women undergoing MIS.


Duodeno-pleural fistula: a rare complication of peptic ulcer perforation.
Duodenopleural fistula is a very uncommon complication of peptic ulcer perforation and usually follows empyema after a subdiaphragmatic abscess rupture. We present a rare case of duodenopleural fistula following subdiaphragmatic abscess, which resulted in thoracic empyema after gastric perforation.


Two forms of hippocampal long-term depression, the counterpart of long-term potentiation.
In the hippocampus there are two distinct forms of long-term depression (LTD) of excitatory synaptic transmission. In the CA1 region, prolonged low-frequency stimulation induces LTD by activating postsynaptic NMDA receptors, which causes a moderate rise in Ca2+ concentrations. In mossy fiber synapses of the CA3 region, similar low-frequency stimulation also gives rise to LTD. However, this form of LTD (mossy fiber LTD) does not require activation of NMDA receptors, but is mediated by activation of presynaptic metabotropic glutamate receptors. Induction of mossy fiber LTD is not dependent on postsynaptic depolarization or activation of postsynaptic ionotropic glutamate receptors, thus it is likely to be mediated by purely presynaptic mechanisms. This conclusion is confirmed by the analysis of mutant mice lacking presynaptic mGluR2, in which mossy fiber LTD is almost absent. Since long-term potentiation at mossy fiber synapses is also induced presynaptically, the synaptic efficacy may be regulated through common mechanisms bidirectionally, which may contribute to neural information processing in the hippocampus.


Anaesthetic Efficacy of Topical Benzocaine Gel Combined with Hyaluronidase for Supplemental Intrapulpal Injection in Teeth with Irreversible Pulpitis- A Double Blinded Clinical Trial.
Intrapulpal injection technique is one of the most commonly employed method to achieve profound pulpal anaesthesia during an endodontic procedure. To determine if the topical application of benzocaine gel along with hyaluronidase to the pulp chamber could reduce the pain felt with the intrapulpal injection technique. Two hundred patients with chronic irreversible pulpitis undergoing endodontic treatment for mandibular first molars in which the primary anaesthetic technique failed were selected and randomly divided into 2 groups. In the control group intrapulpal injection was administered with backpressure. In the experimental group topical application of 20% benzocaine gel mixed with hyaluronidase was done over the exposed pulp following which intrapulpal injection was administered with backpressure. Pain assessment was done on a visual analogue scale. There was statistically significant difference (p<0.001) between the two groups. The mean value in the control group corresponded to the pain perception "strong", whereas that of the experimental group corresponded to the pain perception "weak". Topical application of 20% benzocaine gel mixed with hyaluronidase to the exposed pulp reduces the pain encountered with the intrapulpal injection.


Anti-inflammatory effects of 1,25-dihydroxyvitamin D3 in monocytes cultured in serum from patients with type 2 diabetes mellitus and diabetic nephropathy with uremia via Toll-like receptor 4 and nuclear factor-κB p65.
Type 2 diabetes mellitus (T2DM) and diabetic nephropathy (DN) seriously affect the quality of human life. 1,25-dihydroxyvitamin D3 (VD3) is known to exert anti‑inflammatory and immunomodulatory effects. The present study investigated the effects of VD3 on Toll‑like receptor 4 (TLR4) and nuclear factor (NF)‑κB p65 expression in monocytes from patients with T2DM and investigated the underlying mechanisms. Serum from subjects of the control, T2DM and DN with uremia groups was isolated, and THP‑1 monocytes were cultured in these sera with or without VD3. After treatment with lipopolysaccharide (LPS) and interleukin (IL)‑15, monocytes and culture supernatants were collected. The expression of TLR4, TLR9 and IL‑15 mRNA was detected using reverse‑transcription polymerase chain reaction analysis. Furthermore, the protein expression of TLR4, NF‑κB p65 and inhibitor of NF‑κB (IκB) was determined using western blot analysis. The levels of IL‑6 and monocyte chemotactic protein in culture supernatants were detected using ELISAs. The impact of LPS, IL‑15 and VD3 on the TLR/NF‑κB signaling pathway in the T2DM and DN uremia groups was also investigated. In the T2DM and DN uremia groups, LPS and IL‑15 downregulated the expression of IκB and upregulated levels of proteins, including TLR4, NF‑κB p65, IL‑6 and monocyte chemoattractant protein 1, as well as TLR4 and IL‑15 mRNA. There was no significant difference in TLR9 mRNA protein expression among the three groups. VD3 partially blocked the above effects; However, pre‑treatment with VD3 had no significant effect on TLR4 mRNA expression, protein expression of NF-κB p65 and IκB, or the levels of associated inflammatory cytokines. In conclusion, the present study indicated that anti‑inflammatory effects of VD3 in inflammatory immune responses in T2DM and DN uremia are associated with the TLR4/NF-κB p65 signaling pathway.


[Initial experience with endoscopic subfascial dissection of perforating veins in 100 interventions].
Progress in minimal invasive surgery made us look for a new concept to treat our patients with varicosis, which is one of the most frequent operations in our department. We changed our method from an epifascial ligation to an endoscopic subfascial dissection of the perforating veins (ESDP). This method was developed and first described by Hauer in 1985 [2]. Between March 1992 and May 1993 we treated 85 patients by this method, performing ESDP on 100 legs. In most of the cases the operation is combined with a classic Babcock's procedure, although it can be done isolated. Sectioning of the perforating veins needs special instruments and must be performed in partial deprivation of blood supply, which is produced by a sterile Löfqvist roller cuff [5]. The sectioning of the perforating veins through a small incision in the medial part of the lower leg allows a dissection of all perforating veins in the medial and dorsal sections of the lower leg. The subfascial procedure reduces delayed wound healing, especially in patients with trophic skin disorders. The operation in deprivation of blood supply allows smaller and therefore cosmetically better incisions also for the other subcutaneous varices.


Wounding the messenger: the new economy makes occupational health indicators too good to be true.
The U.S. Bureau of Labor Statistics and workers' compensation insurers reported dramatic drops in rates of occupational injuries and illnesses during the 1990s. The authors argue that far-reaching changes in the 1980s and 1990s, including the rise of precarious employment, falling wages and opportunities, and the creation of a super-vulnerable population of immigrant workers, probably helped create this apparent trend by preventing employees from reporting some injuries and illnesses. Changes in the health care system, including loss of access to health care for growing numbers of workers and increased obstacles to the use of workers' compensation, compounded these effects by preventing the diagnosis and documentation of some occupational injuries and illnesses. Researchers should examine these forces more closely to better understand trends in occupational health.


[Inhibitory effects of sinapine on activity of acetylcholinesterase in cerebral homogenate and blood serum of rats].
The present study investigated the inhibitory effects of Chinese herb component sinapine on activity of acetylcholinesterase (AChE) in cerebral homogenate and blood serum of rats. AChE was prepared from cerebral homogenate and blood serum of rats, respectively. Acetylcholinesterase activity assay kit and Chromatometry were used to detect the AChE activity. Sinapine significantly inhibited AChE activity in vitro, with more effective on cerebral homogenate (IC50 3.66 micromol x L(-1)) than blood serum (IC50 22.1 micromol x L(-1)). Sinapine could significantly inhibit the cerebral AChE activity and may be a promising drug used for prevention and cure of Alzheimer's disease as a cholinesterase inhibitor.


ChimST: An Efficient Spectral Library Search Tool for Peptide Identification from Chimeric Spectra in Data-Dependent Acquisition.
Accurate and sensitive identification of peptides from MS/MS spectra is a very challenging problem in computational shotgun proteomics. To tackle this problem, spectral library search has been one of the competitive solutions. However, most existing library search tools were developed on the basis of one peptide per spectrum, which prevents them from working properly on chimeric spectra where two or more peptides are co-fragmented. In this work, we present a new library search tool called ChimST, which is particularly capable of reliably identifying multiple peptides from a chimeric spectrum. It starts with associating each query MS/MS spectrum with MS precursor features. For each precursor feature, there is a list of peptide candidates extracted from an input spectral library. Then, it takes one peptide candidate from each associated feature and scores how well they could collectively interpret the query spectrum. The highest-scoring set of peptide candidates are finally reported as the identification of the query spectrum. Our experimental tests show that ChimST could significantly outperform the three state-of-the-art library search tools, SpectraST, reSpect, and MSPLIT, in terms of the numbers of both peptide-spectrum matches and unique peptides, especially when the acquisition isolation window is broad.


The potential role of comprehensive genomic profiling to guide targeted therapy for patients with biliary cancer.
Remarkable advancements in techniques of genomic profiling and bioinformatics have led to the accumulation of vast amounts of knowledge on the genomic profiles of biliary tract cancer (BTC). Recent largescale molecular profiling studies have not only highlighted genomic differences characterizing tumors of the intrahepatic and extrahepatic bile ducts and gallbladder, but have also revealed differences in genomic profiles pertaining to associated risk factors. Novel genomic alterations such as FGFR2 fusions and IDH1/2 mutations in intrahepatic cholangiocarcinoma (ICC) and ERBB2 alterations in gallbladder cancer (GBCA) are emerging as targeted therapy options capable of advancing precision medicine for the care of these patients. Moreover, variable genomic alterations also appear to impact prognosis and overall disease outcome independent from their therapy selection value. High mutational burden and increased expression of immune checkpoint-related proteins observed in a subset of BTC also show a potential for guidance of immunotherapy. Thus, comprehensive genomic profiling (CGP) is rapidly achieving status as an integral component of precision medicine and is starting to become invaluable in guiding the management of patients with BTC, a rare disease with dismal outcome.


Nucleosome remodeling by the human SWI/SNF complex requires transient global disruption of histone-DNA interactions.
We utilized a site-specific cross-linking technique to investigate the mechanism of nucleosome remodeling by hSWI/SNF. We found that a single cross-link between H2B and DNA virtually eliminates the accumulation of stably remodeled species as measured by restriction enzyme accessibility assays. However, cross-linking the histone octamer to nucleosomal DNA does not inhibit remodeling as monitored by DNase I digestion assays. Importantly, we found that the restriction enzyme-accessible species can be efficiently cross-linked after remodeling and that the accessible state does not require continued ATP hydrolysis. These results imply that the generation of stable remodeled states requires at least transient disruption of histone-DNA interactions throughout the nucleosome, while hSWI/SNF-catalyzed disruption of just local histone-DNA interactions yields less-stable remodeled states that still display an altered DNase I cleavage pattern. The implications of these results for models of the mechanism of SWI/SNF-catalyzed nucleosome remodeling are discussed.


Functional Analysis of the Ser149/Thr149 Variants of Human Aspartylglucosaminidase and Optimization of the Coding Sequence for Protein Production.
Aspartylglucosaminidase (AGA) is a lysosomal hydrolase that participates in the breakdown of glycoproteins. Defects in the AGA gene result in a lysosomal storage disorder, aspartylglucosaminuria (AGU), that manifests mainly as progressive mental retardation. A number of AGU missense mutations have been identified that result in reduced AGA activity. Human variants that contain either Ser or Thr in position 149 have been described, but it is unknown if this affects AGA processing or activity. Here, we have directly compared the Ser149/Thr149 variants of AGA and show that they do not differ in terms of relative specific activity or processing. Therefore, Thr149 AGA, which is the rare variant, can be considered as a neutral or benign variant. Furthermore, we have here produced codon-optimized versions of these two variants and show that they are expressed at significantly higher levels than AGA with the natural codon-usage. Since optimal AGA expression is of vital importance for both gene therapy and enzyme replacement, our data suggest that use of codon-optimized AGA may be beneficial for these therapy options.


Dual DNA staining enables isolation of multiple sub-types of post-replicative mouse male germ cells.
During spermatogenesis, mammalian male germ cells undergo multiple developmental processes, including meiosis and post-meiotic differentiation (spermiogenesis). To understand the transitions between different cellular states it is essential to isolate pure populations of cells at different stages of development. Previous approaches enabled the isolation of cells from different stages of meiotic prophase I, but techniques to sub-fractionate unfixed, post-meiotic spermatids have been lacking. Here we report the development of a protocol enabling simultaneous isolation of cells at different stages of meiotic prophase and post-meiotic differentiation from testes of adult mice. This approach builds on existing fluorescence activated cell sorting protocols designed to purify cells in different stages of meiotic prophase I. By utilizing the specific spectral properties that two different DNA dyes (Hoechst 33342 and SYTO 16) exhibit when bound to chromatin of different stage male germ cells, we obtain highly pure populations of cells in relatively large numbers. This FACS protocol will enable immunocytological and molecular characterization studies of fractionated meiotic and haploid germ cells from both wild type and genetically mutant animals.


Post-orchidectomy radiation therapy for patients with stage I seminoma of the testis.
The outcome of radiotherapy in patients with stage I testicular seminoma was evaluated. During the period 1960-89 (inclusive) 270 patients with stage I seminoma of the testis received radiotherapy to the para-aortic nodes and ipsilateral hemipelvis following radical orchidectomy. Two hundred and fifty seven patients (95.2%) received a minimum tumour dose of 30Gy in 20 daily fractions using 4-6 MV photons. The 5 year overall and recurrence-free survival rates were 97 and 95%, respectively. Only eight of the 270 patients relapsed and three were cured with 'salvage' therapies. Of the 11 patients who died, four deaths (36%) were the result of uncontrolled testicular cancer, six (55%) intercurrent illness and one (9%) the result of attempted salvage. Patients staged and treated prior to 1979 had a significantly worse disease-free survival compared to patients treated during and after 1979. As side effects were negligible, it was concluded that radiotherapy for stage I seminoma provides excellent cure rates. The difficulties arranging a 'surveillance' programme in so large a State as Queensland are discussed.


Evidence for the involvement of 5f orbitals in the bonding and reactivity of organometallic actinide compounds: thorium(IV) and uranium(IV) bis(hydrazonato) complexes.
Migratory insertion of diphenyldiazomethane into both metal-carbon bonds of the bis(alkyl) and bis(aryl) complexes (C(5)Me(5))(2)AnR(2) yields the first f-element bis(hydrazonato) complexes (C(5)Me(5))(2)An[eta(2)-(N,N')-R-N-N=CPh(2)](2) [An = Th, R = CH(3) (18), PhCH(2) (15), Ph (16); An = U, R = CH(3) (17), PhCH(2) (14)], which have been characterized by a combination of spectroscopy, electrochemistry, and X-ray crystallography. The two hydrazonato ligands adopt an eta(2)-coordination mode leading to 20-electron (for Th) and 22-electron (for U) complexes that have no transition-metal analogues. In fact, reaction of (C(5)H(5))(2)Zr(CH(3))(2) or (C(5)Me(5))(2)Hf(CH(3))(2) with diphenyldiazomethane is limited to the formation of the corresponding mono(hydrazonato) complex (C(5)R(5))(2)M[eta(2)-(N,N')-CH(3)-N-N=CPh(2)](CH(3)) (M = Zr, R = H or M = Hf, R = CH(3)). The difference in the reactivities of the group 4 metal complexes and the actinides was used as a unique platform for investigating in depth the role of 5f orbitals on the reactivity and bonding in actinide organometallic complexes. The electronic structure of the (C(5)H(5))(2)M[eta(2)-(N,N')-CH(3)-N-N=CH(2)](2) (M = Zr, Th, U) model complexes was studied using density functional theory (DFT) calculations and compared to experimental structural, electrochemical, and spectroscopic results. Whereas transition-metal bis(cyclopentadienyl) complexes are known to stabilize three ligands in the metallocene girdle to form saturated (C(5)H(5))(2)ML(3) species, in a bis(hydrazonato) system, a fourth ligand is coordinated to the metal center to give (C(5)H(5))(2)ML(4). DFT calculations have shown that 5f orbitals in the actinide complexes play a crucial role in stabilizing this fourth ligand by stabilizing both the sigma and pi electrons of the two eta(2)-coordinated hydrazonato ligands. In contrast, the stabilization of the hydrazonato ligands was found to be significantly less effective for the putative bis(hydrazonato) zirconium(IV) complex, yielding a higher energy structure. However, the difference in the reactivities of the group 4 metal and actinide complexes does not arise on thermodynamic grounds but is primarily of kinetic origin. Unfavorable steric factors have been ruled out as the sole influence to explain these different behaviors, and electronic factors were shown to govern the reactivity. For the actinides, both the C(5)H(5) and more realistic C(5)Me(5) ligands have been taken into account in computing the energy surface. The reaction profile for the C(5)Me(5) system differs from that with the C(5)H(5) ligand by a uniform shift of approximately 5 kcal/mol in the relative energies of the transition state and products. The insertion of a second diazoalkane molecule into the sole metal-carbon bond in the mono(hydrazonato) complexes involves a high energy barrier (approximately 20 kcal/mol) for the zirconium(IV) system, whereas the actinides can facilitate the approach of the diazoalkane by coordination (formation of an adduct) and its insertion into the An-C bond with a very low barrier on the potential energy surface.


Invited Commentary: When Case-Control Studies Came of Age.
In his 1976 paper "Estimability and Estimation in Case-Referent Studies" (Am J Epidemiol. 1976;103(2):226-235), Miettinen weaved together a patchwork of new ideas into a coherent view of case-control studies. His article spurred theoretical development in epidemiologic methods and became a platform for teaching about some key concepts in epidemiologic study design.


Serum chemerin levels vary with time of day and are modified by obesity and tumor necrosis factor-{alpha}.
Chemerin is an adipokine with important regulatory roles in adipogenesis. In humans, serum total chemerin (i.e. prochemerin plus chemerin) levels are positively associated with body mass index and metabolic syndrome. However, the mechanisms that increase serum chemerin concentration are unknown. We hypothesized that chronic low-grade inflammation that occurs in obesity promotes chemerin production by adipocytes. Consistent with this, TNFalpha treatment of 3T3-L1 adipocytes increased bioactive chemerin levels in the cell media as detected using a CMKLR1 cell-based bioassay. This effect was blocked by the protein synthesis inhibitor cycloheximide and protein secretion inhibitor brefeldin A, indicating that TNFalpha may enhance prochemerin synthesis and secretion from adipocytes. In vivo, TNFalpha produced a time-dependent increase in serum total chemerin and bioactive chemerin. Bioactive chemerin was produced by primary mouse adipocytes and hepatocytes. Only primary adipocyte-derived chemerin was responsive to TNFalpha regulation implicating adipocytes as a potential source of elevated serum chemerin after TNFalpha exposure in vivo. In lean mice, serum total chemerin levels oscillated with peak levels occurring during daytime and trough levels at night. Comparatively, leptin- and leptin receptor-deficient obese mice, which have elevated adipose tissue expression of TNFalpha, displayed elevated serum total chemerin levels with an enhanced oscillatory pattern. In summary, our novel results identified TNFalpha as a positive regulator of adipocyte-derived chemerin. We corroborate the finding of elevated chemerin in obese humans by identifying elevated serum levels of total chemerin in two obese mouse models with a corresponding alteration in the rhythmic pattern of serum chemerin levels.


Neuropilin-1: A feasible link between liver pathologies and COVID-19.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has a tremendous impact on the health of millions of people worldwide. Unfortunately, those suffering from previous pathological conditions are more vulnerable and tend to develop more severe disease upon infection with the new SARS-CoV-2. This coronavirus interacts with the angiotensin-converting enzyme 2 receptor to invade the cells. Recently, another receptor, neuropilin-1 (NRP-1), has been reported to amplify the viral infection. Interestingly, NRP-1 is expressed in nonparenchymal liver cells and is related to and upregulated in a wide variety of liver-related pathologies. It has been observed that SARS-CoV-2 infection promotes liver injury through several pathways that may be influenced by the previous pathological status of the patient and liver expression of NRP-1. Moreover, coronavirus disease 2019 causes an inflammatory cascade called cytokine storm in patients with severe disease. This cytokine storm may influence liver sinusoidal-cell phenotype, facilitating viral invasion. In this review, the shreds of evidence linking NRP-1 with liver pathologies such as hepatocellular carcinoma, liver fibrosis, nonalcoholic fatty liver disease and inflammatory disorders are discussed in the context of SARS-CoV-2 infection. In addition, the involvement of the infection-related cytokine storm in NRP-1 overexpression and the subsequent increased risk of SARS-CoV-2 infection are also analyzed. This review aims to shed some light on the involvement of liver NRP-1 during SARS-CoV-2 infection and emphasizes the possible involvement this receptor with the observed liver damage.


Pharmacokinetic evaluation of liposomal encapsulated ampicillin in male and female rats.
The dispositions of free and liposomal entrapped ampicillin were compared in male and female rats after i.v. administration. Serial blood samples were collected for 2 h in the free drug study and 12 h for the liposomal formulation. Pharmacokinetic parameters obtained with free drug were not significantly different between genders. However, gender significantly influenced the disposition of liposomal encapsulated ampicillin. While no difference was observed in distribution t1/2 between genders, female rats had a shorter MRT, smaller Vss and Vt, and faster clearance as compared to male rats. In a second study, spleen, liver, kidney, heart, and lung were harvested post-injection of free and liposomal entrapped ampicillin. Free ampicillin did not distribute extensively into the tissue compartment and no gender difference was noted. In contrast, liposomal encapsulation resulted in a substantial tissue uptake. In general, female rats had higher concentrations in the spleen and lung as compared to male rats. In vitro plasma stability was not significantly different, suggesting that destabilization of the liposomes does not play a large role in the dispositional differences observed in these studies. However, in vivo interaction of liposomes and plasma lipoproteins may influence the disposition of encapsulated drug.


[Intra-aortic balloon pumping and heparin-induced thrombocytopenia].
Thrombocytopenia is frequently reported in the presence of intra-aortic balloon pumping (IABP) after cardiac surgery and in cardiology, but heparin-induced thrombocytopenia (HIT) is rarely responsible of it in this circumstance. A case-report if HIT in a patient with IABP is presented. This case emphasised the difficulty in diagnosis of HIT in this situation. An acute decrease in platelet count lasting after removal of IABP could suggest the diagnosis of HIT when other origins for thrombocytopenia are excluded.


A newly identified member of the tumor necrosis factor receptor superfamily with a wide tissue distribution and involvement in lymphocyte activation.
The tumor necrosis factor receptor (TNFR) superfamily consists of approximately 10 characterized members of human proteins. We have identified a new member of the TNFR superfamily, TR2, from a search of an expressed sequence tag data base. cDNA cloning and Northern blot hybridization demonstrated multiple mRNA species, of which a 1.7-kilobase form was most abundant. However, TR2 is encoded by a single gene which, maps to chromosome 1p36.22-36.3, in the same region as several other members of the TNFR superfamily. The most abundant TR2 open reading frame encodes a 283-amino acid single transmembrane protein with a 36-residue signal sequence, two perfect and two imperfect TNFR-like cysteine-rich domains, and a short cytoplasmic tail with some similarity to 4-1BB and CD40. TR2 mRNA is expressed in multiple human tissues and cell lines and shows a constitutive and relatively high expression in peripheral blood T cells, B cells, and monocytes. A TR2-Fc fusion protein inhibited a mixed lymphocyte reaction-mediated proliferation suggesting that the receptor and/or its ligand play a role in T cell stimulation.


Depression, anxiety, insomnia and interleukins in the early postpartum period.
Some new mothers have been shown to suffer from anxiety and depression associated with insomnia during the postpartum period. Our study assessed the impact of demographic, psychopathological, and biochemical factors on the incidence of depression in women during the early postpartum period. A total of 119 women were evaluated at 24-48 h postpartum with the following psychometric scales: Hamilton Depression Rating Scale (HDRS), Edinburgh Postnatal Depression Scale (EPDS), Hamilton Anxiety Rating Scale (HARS) and Athens Insomnia Scale (AIS). In addition, blood was drawn to assay interleukin 6 (IL-6) and interleukin 10 (IL-10). The factors that had the greatest impact on the risk of postpartum depression detected with the HDRS were high HARS scores and evidence of insomnia in the AIS. There were no significant differences in IL-6 or IL-10 levels in women with and without depression (based on either HDRS or EPDS scores) and insomnia (based on AIS) after childbirth. Considering demographic factors, divorced and single women were shown to be at higher risk of postpartum depression (based on EPDS scores). Small sample size and short observation span. This study highlights the relationship between postpartum depression and both anxiety and insomnia and emphasises the importance to assess symptoms of anxiety and sleep quality as part of screening in women at risk of postpartum depression.


High-resolution cytogenetic analysis of the peripheral blood lymphocytes of patients with autosomal dominant polycystic kidney disease.
High-resolution cytogenetic analysis was performed on peripheral blood lymphocytes of 11 patients (seven women and four men, range 24-56 years) with proven autosomal dominant polycystic kidney disease (ADPKD). A total of 449 metaphases (an average of 41 per patient) were analyzed. In two of the women changes in the karyotype were found in a single cell: del(3), (q21) in one of them and Int.del(6), (q11q21) in the other. Analysis of the remaining patients, including those in which ADPKD was combined with oligophrenia, did not reveal any deviations from the normal karyotype. Therefore, the established chromosome changes are non-specific and of little importance in diagnosing ADPKD.


A molecular mechanism for qE-quenching.
We discuss energy-dependent fluorescence lowering (qE-quenching), and suggest a model to explain the experimental data currently available. The main elements of the model are: (a) the qE-quenching reflects a mechanism associated with a component of the light-harvesting antenna rather than the reaction center of photosystem (PS) II--we suggest that it occurs through formation of an efficient quencher in one of the minor chlorophyll protein (CP) complexes; (b) the minor CPs have glutamate residues instead of glutamines at positions shown in light-harvesting complex II (LHCII) to be ligands to chlorophylls near the lumenal interface. We suggest that the quenching reflects a change in ligation of chlorophyll on protonation of these glutamate residues leading to formation of an exciton coupled dimer with a neighboring pigment, in which additional energy levels allow vibrational relaxation of the excited singlet. The model accounts for the dependence on low lumenal pH, the ligand residue changes between LHCII and the minor CPs, the preferential distribution of components of the xanthophyll cycle in the minor CPs, the inhibition of qE-quenching by DCCD, and the specific binding of DCCD to the minor CPs.


The relationship between dietary supplement use in late pregnancy and birth outcomes: a cohort study in British women.
To examine the relationship between dietary supplement use during pregnancy and birth outcomes. A prospective birth cohort. Leeds, UK. One thousand two hundred and seventy-four pregnant women aged 18-45 years. Dietary supplement intake was ascertained using three questionnaires for the first, second and third trimesters. Dietary intake was reported in a 24-hour dietary recall administered by a research midwife at 8-12 weeks of gestation. Information on delivery details and antenatal pregnancy complications was obtained from the hospital maternity records. Birthweight, birth centile and preterm birth. Reported dietary supplement use declined from 82% of women in the first trimester of pregnancy to 22% in the second trimester and 33% in the third trimester. Folic acid was the most commonly reported supplement taken. Taking any type of daily supplement during any trimester was not significantly associated with size at birth taking into account known relevant confounders. Women taking multivitamin-mineral supplements in the third trimester were more likely to experience preterm birth (adjusted OR = 3.4, 95% CI 1.2, 9.6, P = 0.02). Regular multivitamin-mineral supplement use during pregnancy, in a developed country setting, is not associated with size at birth. However, it appears to be associated with preterm birth if taken daily in the third trimester. The mechanism for this is unclear and our study's findings need confirming by other cohorts and/or trials in developed countries.


Novel geometries for tissue-engineered tendonous collagen constructs.
A promising approach to addressing the performance limitations of currently available mechanical and bioprosthetic heart valves lies in tissue engineering. Tissue-engineered valves should incorporate the complex microstructure of the native valves to mimic their unique mechanics. This would include a layered topology, mesh networks, and branched collagen fiber bundles. Our approach to heart valve tissue engineering is to develop the functional components of the aortic valve cusps separately in vitro and, once they are mature, integrate them into a composite valve structure. Here we report on our efforts to create more complex collagenous structures, suitable for heart valve tissue engineering. Collagen fiber bundles were fabricated using the principle of directed collagen gel contraction, using neonatal rat aortic smooth muscle cells and acid-soluble type I rat-tail tendon collagen. The collagen gels were cast into rectangular or branched wells with porous end holders that constrained the gels longitudinally but allowed contraction to occur transverse to the long axis. Pairs of such constructs were placed in direct contact with each other and cultured further to determine whether they integrated to form continuous tissue. After 6-8 weeks of culture, highly compacted and aligned collagen fiber bundles formed. Mechanical testing revealed that linear constructs (2 free ends) with an 8:1 aspect ratio were significantly stronger than similar constructs with an aspect ratio of 2:1 (mean +/- SD, 298 +/- 90 kPa vs. 152 +/- 49 kPa; p < .001). Branching reduced mechanical strength considerably. Constructs fabricated with 4 free ends were significantly weaker than constructs with 3 ends (31 +/- 32 kPa vs. 116 +/- 66 kPa; p < .003). Histologic images demonstrated the integration of the crossed collagen bundles, with a bonding strength of 2.1 +/- 1.1 g (0.02 N). We found that the geometry of the molds into which the collagen constructs are cast can greatly affect their mechanical strength: multibranched constructs were the weakest, and long, linear constructs were the strongest. We also found that integration of collagen constructs occurs in vitro and that the fabrication of a composite structure in vitro is probably feasible.


BRAF as therapeutic target in melanoma.
BRAF is a member of the RAF kinase family, which acts in the ERK/MAP kinase pathway, a signalling cascade that regulates cellular proliferation, differentiation and survival. Single point mutations can turn BRAF into an oncogene, but there appears to be a cell type/tumour specific relevance for BRAF kinase-activating mutations, since they are found predominantly in cutaneous melanoma. With the success of targeting other oncogenic kinases such as BCR-ABL, KIT or members of the epidermal-growth factor receptor (EGFR) family in other cancers, the expectations were high when the first RAF kinase-targeting drug (sorafenib) reached clinical trials. However, disappointingly the first studies using sorafenib in melanoma patients did not show the anticipated single agent efficacy. More recently, the resolution of the BRAF crystal structure has led to the development of better, more specific BRAF inhibitors such as the Plexxikon compound, PLX4032, which induced a dramatic response rate in phase I trials, validating BRAF as a clinically relevant target. In addition, our understanding of melanoma biology and the role BRAF is playing therein has improved significantly. The complexity in the ERK/MAP kinase pathway including important feedback mechanisms has been dissected, and the relevance of cross-talks with other signalling pathways has been revealed, suggesting strategies for the design of improved, more efficient combinatorial therapies. This review highlights the relevance of BRAF and the ERK/MAP kinase pathway for melanoma cell biology and discusses some of the recent advances in both, the understanding of BRAF function in melanoma and the development of improved BRAF targeting inhibitors.


Tapeworms (Cestoda: Proteocephalidae) of Australian reptiles: hidden diversity of strictly host-specific parasites.
Reptilian fauna of Australia is extraordinarily rich and diverse, but very little is known about parasites of reptiles, including proteocephalid cestodes of the genus Ophiotaenia La Rue, 1911. In the present survey, data on these parasites are summarised for the first time based on detailed evaluation of all available type and voucher specimens. This survey includes five named species, with four species redescribed, namely O. amphiboluri (Nybelin, 1917), O. longmani Johnston, 1916, O. mjobergi (Nybelin, 1917), and O. striata (Johnston, 1914), and 9 putative new species which are not formally described because of their poor quality and scarce material. An identification key is given for the five named species and unnamed species are briefly characterised with focus on their differential characteristics to facilitate their description as new taxa in the future. Australian species of Ophiotaenia form a monophyletic lineage, whose members share several morphological traits which are absent or rare in other proteocephalids, such as a three-layered embryophore, a scolex with large, anteriorly directed suckers, eggs with thick-walled embryophores, exclusively dorsal and paramuscular position of vitelline follicles, and a postequatorial to equatorial genital pore in most species. A new genus, Australophiotaenia, is proposed to accommodate the species from Australian reptiles that share the above-mentioned characters.


Reevaluation of antimicrobial and antioxidant activity of Thymus spp. extracts before and after encapsulation in liposomes.
The antioxidant and antimicrobial activity of four Thymus species (boissieri, longicaulis, leucospermus, and ocheus) extracts were determined. Two methods (Rancimat and malondialdehyde by high-performance liquid chromatography) were used to measure the antioxidant action in comparison with common commercial antioxidants, including butylated hydroxytoluene and alpha-tocopherol. The extracts that presented high antioxidant activity were encapsulated in liposomes and their antioxidant action was again estimated. Thermal-oxidative decomposition of the samples (pure liposomes and encapsulating extracts) was studied using the differential scanning calorimetry method. The modification of the main transition temperature for the lipid mixture and the splitting of the calorimetric peak in the presence of the antioxidants were also demonstrated by differential scanning calorimetry. All extracts showed antioxidant and antimicrobial activities. Some extracts showed superior or equal antioxidant activity to alpha-tocopherol. When the extracts were encapsulated in liposomes, their antioxidant as well as antimicrobial activities proved to be superior from the same extracts in pure form.


Isolation and characterization of a virus infecting the freshwater algae Chrysochromulina parva.
Water samples from Lake Ontario, Canada were tested for lytic activity against the freshwater haptophyte algae Chrysochromulina parva. A filterable lytic agent was isolated and identified as a virus via transmission electron microscopy and molecular methods. The virus, CpV-BQ1, is icosahedral, ca. 145nm in diameter, assembled within the cytoplasm, and has a genome size of ca. 485kb. Sequences obtained through PCR-amplification of DNA polymerase (polB) genes clustered among sequences from the family Phycodnaviridae, whereas major capsid protein (MCP) sequences clustered among sequences from either the Phycodnaviridae or Mimiviridae. Based on quantitative molecular assays, C. parva׳s abundance in Lake Ontario was relatively stable, yet CpV-BQ1׳s abundance was variable suggesting complex virus-host dynamics. This study demonstrates that CpV-BQ1 is a member of the proposed order Megavirales with characteristics of both phycodnaviruses and mimiviruses indicating that, in addition to its complex ecological dynamics, it also has a complex evolutionary history.