Alkaline brain pH shift in rodent lithium-pilocarpine model of epilepsy with chronic seizures.
Brain pH is thought to be important in epilepsy. The regulation of brain pH is, however, still poorly understood in animal models of chronic seizures (SZ) as well as in patients with intractable epilepsy. We used chemical exchange saturation transfer (CEST) MRI to noninvasively determine if the pH is alkaline shifted in a rodent model of the mesial temporal lobe (MTL) epilepsy with chronic SZ. Taking advantage of its high spatial resolution, we determined the pH values in specific brain regions believed to be important in this model produced by lithium-pilocarpine injection. All animals developed status epilepticus within 90 min after the lithium-pilocarpine administration, but one animal died within 24 hrs. All the surviving animals developed chronic SZ during the first 2 months. After SZ developed, brain pH was determined in the pilocarpine and control groups (n = 8 each). Epileptiform activity was documented in six pilocarpine rats with scalp EEG. The brain pH was estimated using two methods based on magnetization transfer asymmetry and amide proton transfer ratio. The pH was alkaline shifted in the pilocarpine rats (one outlier excluded) compared to the controls in the hippocampus (7.29 vs 7.17, t-test, p < 0.03) and the piriform cortex (7.34 vs. 7.06, p < 0.005), marginally more alkaline in the thalamus (7.13 vs. 7.01, p < 0.05), but not in the cerebral cortex (7.18 vs. 7.08, p > 0.05). Normalizing the brain pH may lead to an effective non-surgical method for treating intractable epilepsy as it is known that SZ can be eliminated by lowering the pH.


Albumin therapy of transient focal cerebral ischemia: in vivo analysis of dynamic microvascular responses.
To study whether intravascular or hemodynamic factors contribute to the marked neuroprotective effect of albumin therapy in focal cerebral ischemia, 2 complementary methods were applied: laser-scanning confocal microscopy (LSCM) and laser-Doppler perfusion imaging (LDPI). In the LSCM study, Sprague-Dawley rats were anesthetized with halothane/nitrous oxide, and a cranial window was placed over the dorsolateral frontoparietal cortex. Rats received 2-hour middle cerebral artery occlusion (MCAO) by an intraluminal suture and were treated with human albumin (1.25 g/kg; n=4) or saline (n=3) after 30 minutes of recirculation. Video images of cortical vessels were continually acquired and were digitized offline to measure diameters and fluorescent erythrocyte velocities. In the LDPI study, cortical perfusion was measured in anesthetized Sprague-Dawley rats that received 2-hour MCAO and were treated with albumin (2.5 g/kg; n=6) or saline (n=5) at 30 minutes after recirculation. In the LSCM study, MCAO was associated with arteriolar dilation and slowing of capillary and venular erythrocyte perfusion. During the first 15 to 30 minutes of postischemic recirculation, prominent foci of vascular stagnation developed within cortical venules, associated with thrombuslike foci and adherent corpuscular structures consistent in size with neutrophils. Saline administration failed to affect these phenomena, while albumin therapy was followed by significant increases in arteriolar diameter ( approximately 12%; P=0.007) and by a prompt improvement of venular and capillary erythrocyte perfusion and a partial disappearance of adherent thrombotic material. Albumin therapy increased erythrocyte flow velocity in both capillaries (288+/-73% versus 76+/-18% in the saline group; P=0.023) and venules (2.7-fold [P=0.001] versus 1.0-fold in the saline group [P=NS]). In the LDPI study, cortical perfusion declined during MCAO and rose initially with recirculation (to approximately 135% of baseline) in both groups. Mean cortical perfusion improved slightly (approximately 14%; P=NS) in albumin-treated animals. These results reveal a beneficial effect of albumin therapy in reversing stagnation, thrombosis, and corpuscular adherence within cortical venules in the reperfusion phase after focal ischemia and support its utility in the treatment of acute ischemic stroke.


Recovery of metallo-tolerant and antibiotic resistant psychrophilic bacteria from Siachen glacier, Pakistan.
Cultureable bacterial diversity of previously unexplored Siachen glacier, Pakistan, was studied. Out of 50 isolates 33 (66%) were Gram negative and 17 (34%) Gram positive. About half of the isolates were pigment producers and were able to grow at 4-37°C. 16S rRNA gene sequences revealed Gram negative bacteria dominated by Proteobacteria (especially γ-proteobacteria and β-proteobacteria) and Flavobacteria. The genus Pseudomonas (51.51%, 17) was dominant among γ- proteobacteria. β-proteobacteria constituted 4 (12.12%) Alcaligenes and 4 (12.12%) Janthinobacterium strains. Among Gram positive bacteria, phylum Actinobacteria, Rhodococcus (23.52%, 4) and Arthrobacter (23.52%, 4) were the dominating genra. Other bacteria belonged to Phylum Firmicutes with representative genus Carnobacterium (11.76%, 2) and 4 isolates represented 4 genera Bacillus, Lysinibacillus, Staphylococcus and Planomicrobium. Most of the Gram negative bacteria were moderate halophiles, while most of the Gram positives were extreme halophiles and were able to grow up to 6.12 M of NaCl. More than 2/3 of the isolates showed antimicrobial activity against multidrug resistant S. aureus, E. coli, Klebsiella pneumonia, Enterococcus faecium, Candida albicans, Aspergillus flavus and Aspergillus fumigatus and ATCC strains. Gram positive bacteria (94.11%) were more resistant to heavy metals as compared to Gram negative (78.79%) and showed maximum tolerance against iron and least tolerance against mercury.


[Outcomes of implanting porcine small intestinal submucosa mesh in rabbit vesicovaginal space].
Objective: To assess surgical outcomes of implanted porcine small intestinal submucosa (SIS) mesh in the rabbit vesicovaginal space (VVS) and explore its application value in pelvic floor reconstruction surgery. Methods: Sixteen male rabbits were randomly divided into four groups, and each group had four rabbits. All groups of rabbits were implanted with SIS mesh in the vesicovaginal space. They were humanely killed after a postoperative period of 7, 30, 90 and 180 days by group. The grafted area was removed with the surrounding bladder and vaginal tissues. The specimens were embedded in paraffin and then stained with HE and Masson's trichrome stains for visual observations, cells counts, and assessment of tissues and collagen fibers. Results: (1) After HE staining, a large number of inflammatory response cells mainly eosinophils and lymphocytes infiltrated around the SIS mesh in 7 days group, and neovascularization was observed, the infiltration area of inflammatory response cells further increased in 30 days group, the infiltration area of inflammatory response cells significantly reduced in 90 days group, while the inflammatory response basically subsided in 180 days group. (2) After Masson's trichromestaining, the collagen structure of SIS mesh in 7 days group was clear and intact. While, the collagen structure of SIS mesh was partially degraded in 30 days group, the SIS meshes of 4 rabbits were completely degraded, but the collagen fragments of SIS remained in 90 days group. In 180 days group, the SIS mesh of all rabbits was degraded, and one of them had the formation of new collagen fibers. Conclusions: SIS mesh implanted into the VVS of rabbits can lead to a transient non infective inflammatory reaction, which could be completely degraded and a small amount of new collagen fibers could be produced after 180 days of implantation. Which shown that SIS mesh should be used cautiously in pelvic floor reconstruction surgery.


Relevant Features of a Triethylene Glycol Dimethyl Ether-Based Electrolyte for Application in Lithium Battery.
Triethylene glycol dimethyl ether (TREGDME) dissolving lithium trifluoromethanesulfonate (LiCF3SO3) is studied as a suitable electrolyte medium for lithium battery. Thermal and rheological characteristics, transport properties of the dissolved species, and the electrochemical behavior in lithium cell represent the most relevant investigated properties of the new electrolyte. The self-diffusion coefficients, the lithium transference numbers, the ionic conductivity, and the ion association degree of the solution are determined by pulse field gradient nuclear magnetic resonance and electrochemical impedance spectroscopy. The study sheds light on the determinant role of the lithium nitrate (LiNO3) addition for allowing cell operation by improving the electrode/electrolyte interfaces and widening the voltage stability window. Accordingly, an electrochemical activation procedure of the Li/LiFePO4 cell using the upgraded electrolyte leads to the formation of stable interfaces at the electrodes surface as clearly evidenced by cyclic voltammetry, impedance spectroscopy, and ex situ scanning electron microscopy. Therefore, the lithium battery employing the TREGDME-LiCF3SO3-LiNO3 solution shows a stable galvanostatic cycling, a high efficiency, and a notable rate capability upon the electrochemical conditions adopted herein.


Quantitative prediction of oral cancer risk in patients with oral leukoplakia.
Exfoliative cytology has been widely used for early diagnosis of oral squamous cell carcinoma. We have developed an oral cancer risk index using DNA index value to quantitatively assess cancer risk in patients with oral leukoplakia, but with limited success. In order to improve the performance of the risk index, we collected exfoliative cytology, histopathology, and clinical follow-up data from two independent cohorts of normal, leukoplakia and cancer subjects (training set and validation set). Peaks were defined on the basis of first derivatives with positives, and modern machine learning techniques were utilized to build statistical prediction models on the reconstructed data. Random forest was found to be the best model with high sensitivity (100%) and specificity (99.2%). Using the Peaks-Random Forest model, we constructed an index (OCRI2) as a quantitative measurement of cancer risk. Among 11 leukoplakia patients with an OCRI2 over 0.5, 4 (36.4%) developed cancer during follow-up (23 ± 20 months), whereas 3 (5.3%) of 57 leukoplakia patients with an OCRI2 less than 0.5 developed cancer (32 ± 31 months). OCRI2 is better than other methods in predicting oral squamous cell carcinoma during follow-up. In conclusion, we have developed an exfoliative cytology-based method for quantitative prediction of cancer risk in patients with oral leukoplakia.


Multiple Gastrointestinal Cancers in a Single Patient-a Rare Clinical Entity.
Multiple gastrointestinal cancers in a single patient is a rare entity. In our study, we are showing the clinical presentation and management of these patients. A fifty-nine-year-old asthenic male (already treated case of metachronous colorectal cancer in 2008 and 2011) presented with complaints of generalized weakness and fatigue. Strong family history was present with two of his first-degree relatives having diagnosed with gastrointestinal cancer at the age < 50 years with one of them having stomach carcinoma and another with GEJ tumors. On evaluation, upper GI endoscopy revealed growth at cardia and endoscopic biopsy revealed adenocarcinoma. Radiological evaluation with PET-CT scan revealed proximal stomach growth with regional lymphadenopathy. Patient was optimized for surgery and underwent D2 total gastrectomy, distal pancreatectomy and splenectomy with Roux-en-Y oesophago-jejunal anastomosis. Pathological stage revealed pT4N2M0, moderately differentiated adenocarcinoma of proximal stomach, both distal and proximal cut margins negative for tumor, LVI present with no perineural invasion, and 5/18 lymph nodes dissected were positive for malignancy. Genetic testing needs to be considered in this patient (modified Bethesda guidelines and IGCLC criteria). Familial gastric cancer are of two types: (a) hereditary diffuse gastric cancer syndrome, (b) familial intestinal type gastric cancer. Approximately 5% of patients have germ-line mutations-AD LYNCH syndrome, hereditary breast-ovarian cancer, and polyposis and non-polyposis syndrome. Once diagnosed in localized advanced stage, the best treatment is R0 resection though overall prognosis in these patients is very poor. So it is rationale to find such families with elevated risk and to do active surveillance for early diagnosis and providing prophylactic gastrectomies to them as it has proven to be beneficial in hereditary form of gastric cancer.


[The palatine tonsils and immunity. Report VII. Soluble factors produced by the lymphocytes of the palatine tonsils in vitro].
A study was made of the production of a blastogenic factor and lymphotoxin in the cultures of lymphocytes of the palatine tonsils removed from patients with chronic tonsillitis; the activity of this blastogenic factor and lymphotoxin was studied in the test-cultures of autologous and allogenic lymphocytes and the transplantable HeLa cells. The antigens of the pathogenic streptococcus and staphylococcus induced production of the blastogenic factor and lymphotoxin; as to the antigens of saprophytic bacteria-they produced no such action. The antigen-specific blastogenic factor intensified the immune response to the homologous antigen, whereas the factor obtained in stimulation of lymphocytes with PHA-to all the microbial antigens under study.


Squamous cell carcinoma of the vulva: study of ploidy, HPV, p53, and pRb.
Seventy-one cases of invasive squamous cell carcinoma (ISCC) of the vulva were compared with 18 cases of vulvar intraepithelial neoplasia (VIN) and 21 cases of lichen sclerosus. Ploidy was studied by image analysis, HPV-DNA by PCR, and p53 and pRb by immunohistochemistry. Univariate and multivariate statistical analyses were performed. The mean age of the patients with ISCC was 70.6 years; only 8.5% were < 60 (range, 43-89) years. For the 43 patients with follow-up, FIGO surgical stages were I in 16.2%, II in 48.8%, III in 27.9%, and IV in 6.9%. The 5-year survival was 90% for the patients with curative surgery (vulvectomy and lymphadenectomy) and 32% for those with tumors in stages III to IV. Previous history of nonneoplastic epithelial alterations was recorded in 54%. Vascular invasion was detected in 4.3% and perineural invasion in 21.4%. Inguinal lymph node metastases were present in 34.9% of the cases. Fifty-one (72%) ISCCs were aneuploid, HPV-DNA-16 was detected in 7 (12.3%) cases, overexpression of p53 was found in 40 (56%), and pRb expression was negative in 15 (21.4%). Fifteen cases (80%) of VIN were aneuploid, 5 (27.7%) contained HPV-DNA, 11 (61%) were positive for p53, and all immunoreacted for pRb. All lichen sclerosus cases were diploid, did not contain HPV-DNA, failed to stain for p53, and were positive for pRb. Our study confirmed the prognostic value of conventional pathological features: stage, lymph node metastasis, histological grade, and vascular and perineural invasion; all were statistically significant for survival in the univariate analysis. Also, ploidy was significant in patients with stages I and II tumors. The only significant variable in the multivariate analysis was stage. p53 overexpression appears as a late event in vulvar carcinogenesis, but it may occur before tumor invasion. Lack of pRb expression can occur in vulvar neoplasia, but it does not seem to play any role in the initiation or prognosis of vulvar ISCC.


Birth outcome, not pregnancy process: reply to van der Veen.
In a recent article (Frisbie, Forbes, and Pullum 1996) we documented racial/ethnic differences in birth outcomes according to a more fine-grained classification than has typically been employed in the demographic literature. In his commentary, van der Veen focuses on the measurement of one of the dimensions of that classification, maturity of the infant, as proxied by the fetal growth ratio. The crux of the critique is easily seen in van der Veen's statement that "all of my disagreements with Frisbie et al.'s method arise from their particular use of a postnatal standard for the assessment of intrauterine growth." Our critic misunderstands our objective: He fails to realize our interest in birth outcome, not pregnancy process, and does not perceive that our intent was to extend the research extant in both the demographic and public-health literatures in which patently postnatal (i.e., ex utero) measures are taken as outcomes interesting in their own right and/or as risk factors for infant mortality and infant and childhood morbidity. Specifically, he does recognize that we purposefully expanded our focus to include moderately compromised births to determine if they were at higher risk than the normal births with whom they are conventionally categorized. Our discussion draws on research cited in the original article, on studies cited by our critic, and on a few more recent investigations. Although we have never argued that ours is the only, or even the best approach in all cases, we try to clarify the rationale for, and adduce additional empirical evidence of, the utility of the method we used.


Conservation programmes for African cattle: design, cost and benefits.
Information on costs and benefits of conservation programmes for cattle is scarce in the literature and mainly available for Europe. This study aims at simulating cost of and benefits from different kinds of conservation programmes designed for indigenous African cattle. The programmes include installation of a herdbook and activities to promote the breed (HB), in situ conservation with a sire rotation scheme (IS), cryoconservation of semen (CC) and CC combined with in situ conservation (IC). The results indicate that cost of the analysed conservation programmes was generally higher than those reported in the literature for comparable schemes. If cost per effective population size conserved is considered, programmes analysed in this study do not appear to be more expensive. The proposed rotation scheme in IS can be applied to many different production systems and prove to be effective with regard to low increase in kinship. Reduction in extinction probability is found to be higher for conservation programmes that strongly involve farmers and give them part of the responsibility for the breeding population. IC was most efficient with regard to cost per effective population size conserved. However, if cost per reduction in endangerment is considered as criterion for the efficiency of a programme, IS, HB and CC are superior to IC. These findings suggest that decisions on conservation programmes should be based on multiple criteria, and not just on cost per effective population size.


Modulation transfer function of fluorescent screens excited by x-rays.
A theoretically based calculation of the modulation transfer function is performed for phosphor screens excited by x-rays. It is assumed that the screens are of compact material so that scattering of the light generated can be neglected. The absorption of both x-rays and light in the phosphor material is taken into account.


Imaging of bone and joint infections.
In recent years, world interest in bone infection has centered on investigating the potential of the newer imaging methods for detecting early osteomyelitis and on their sensitivity and specificity in evaluating both the acute disease and recurrence of chronic disease and the development of infection in prostheses. No revolutionary new techniques were forthcoming since the advent of MR imaging, and the past 12 months have seen a decline in new analyses, presumably because most researchers have now made up their minds about the relative imaging modalities. Interest lately has been mainly in the imaging of chronic (granulomatous) disease and patterns in children and in drug abuse patients. Because of the importance of differentiating neuropathy from infection in the diabetic foot, research is also active in this field, particularly in relation to dynamic scintigraphy. Interesting observations continue to emerge on infection resulting from the depressing spread of drug abuse and immunosuppression.


Patient mobility in European Union: health spas in Ischia, Italy.
In a new case on patients seeking medical services abroad, the Leichtle case, the European Court of Justice (ECJ) confirmed its previous rulings on patient mobility. According to the Court, patients in the European Union have a (conditional) right to receive health care abroad, whereas the sickness fund should reimburse the costs of treatment and travel expenditures. As such, the Court has strengthened patient mobility in the European Union, based on the free movement principles. Now, it is up to the European Commission to develop a communal strategy aimed at further strengthening patients' rights in the Union.


Adnexal surgery in dogs and cats.
Abnormalities in eyelid position and function can lead to chronic irritation of cornea and conjunctiva. Corneal ulceration, vascularization and pigmentation are common consequences. Loss of vision, or loss of the globe may be the end result. The combination of ptosis and entropion of the upper eyelid is commonly seen in breeds with abundant forehead skin folds. Treatment options include surgical reduction of skin folds, suspending the brows, enforced secondary granulation of the upper eyelid, or a combination of procedures. Lid-shortening procedures reduce corneal exposure, improve functional blinking and reduce likelihood of traumatic proptosis. Although a lateral canthoplasty is technically easier to perform, a medial canthoplasty is the preferred method of eyelid shortening. An involuted lateral canthus with entropion is seen in certain dog breeds with a broad skull base and redundant facial skin. Surgical correction is by either applying traction to the lateral canthus, or by resection of the lateral canthal ligament. A combination of entropion, ectropion and macroblepharon may result in the presence of diamond shaped eyelids. Surgical correction involves the combination of several procedures. Upper eyelid agenesis is a congenital absence of part of the upper eyelid in cats. Treatment options include a skin-orbicularis flap from the lower eyelid with a conjunctival pedical graft, the transposition of the lower eyelid to the upper eyelid, cross lid flap and subdermal plexus flap, and subdermal collagen injection. Several techniques for reconstruction of upper and lower eyelid defects are described.


Warts--a clinical's view.
We do have more modalities for treating warts than ever before, but the very plurality of approaches bespeaks the fact that no single method is adequate. We await the development of a safe and effective wart vaccine with keen anticipation.


Dose equations for tube current modulation in CT scanning and the interpretation of the associated CTDIvol.
The scanner-reported CTDI(vol) for automatic tube current modulation (TCM) has a different physical meaning from the traditional CTDI(vol) at constant mA, resulting in the dichotomy "CTDI(vol) of the first and second kinds" for which a physical interpretation is sought in hopes of establishing some commonality between the two. Rigorous equations are derived to describe the accumulated dose distributions for TCM. A comparison with formulae for scanner-reported CTDI(vol) clearly identifies the source of their differences. Graphical dose simulations are also provided for a variety of TCM tube current distributions (including constant mA), all having the same scanner-reported CTDI(vol). These convolution equations and simulations show that the local dose at z depends only weakly on the local tube current i(z) due to the strong influence of scatter from all other locations along z, and that the "local CTDI(vol)(z)" does not represent a local dose but rather only a relative i(z) ≡ mA(z). TCM is a shift-variant technique to which the CTDI-paradigm does not apply and its application to TCM leads to a CTDI(vol) of the second kind which lacks relevance. While the traditional CTDI(vol) at constant mA conveys useful information (the peak dose at the center of the scan length), CTDI(vol) of the second kind conveys no useful information about the associated TCM dose distribution it purportedly represents and its physical interpretation remains elusive. On the other hand, the total energy absorbed E ("integral dose") as well as its surrogate DLP remain robust between variable i(z) TCM and constant current i0 techniques, both depending only on the total mAs = {i}t0 = i0t0 during the beam-on time t0.


Intraoperative ultrasound for gynecologic procedures.
Recent technological advances in portable dynamic imaging equipment allow for the performance of intraoperative ultrasound examinations during certain gynecologic procedures. Reported is the authors' technique for such an examination and outlined are specific vaginal and abdominal gynecologic procedures wherein adjunctive ultrasound may facilitate performance of the operative procedure, enhance safety, and decrease the possibility of complications.


Predictors of weight reduction and maintenance in a large cohort of overweight and obese adults in a community setting.
In overweight/obese individuals, modest 5% weight loss and its maintenance promotes health; however, it is challenging and typically unachievable, especially in community settings. Established predictors of weight loss outcome in a community setting are lacking, hindering the development of practical interventions and prevention tools. Our aim was to identify predictors associated with 5% weight reduction (initial 6 months) and maintenance (1 year) in overweight and obese adults undergoing weight reduction treatment, free of charge, in a community setting. Computerised medical files of 11 842 adults aged 50.6 ± 16.3 years, attending 162 primary clinics, were analysed retrospectively. Thirty medical, biochemical and demographic independent variables were tested as potential predictors using multiple logistic regression models. Significant predictors of high successful weight reduction were: not being treated with insulin (odds ratio (OR) = 0.53), higher baseline body mass index (OR = 1.05) and younger age (OR = 0.98). Weight maintenance predictors were: successful initial weight reduction (OR = 1.26), short time intervals between weighings (OR = 0.88) and frequent weighing (OR = 0.95). Visits to a dietitian were significantly associated with success during both periods: each visit raised the probability of success by 13.4 and 7.6%, respectively. Type 2 diabetes or use of hypoglycaemic drugs were not significant predictors. In a community-based setting, number of visits to a dietitian is a strong predictor of successful weight reduction and maintenance. Initial success is a critical predictor of weight loss maintenance. Subgroup of older, diabetic patients treated with insulin has a dramatically lower probability of weight reduction success.


Reverse transcriptase of mouse mammary tumour virus: expression in bacteria, purification and biochemical characterization.
We have constructed a plasmid that induces in bacteria the synthesis of an enzymically active reverse transcriptase (RT) of mouse mammary tumour virus (MMTV), a retrovirus with a typical B-type morphology. The highest catalytic activity was detected only when 27 residues from the C-terminus of the protease were included in the N-terminus of the recombinant RT, after an extra deoxyadenosine was added between the pro and pol genes to overcome the -1 frameshift event (which occurs naturally in virus-infected cells). The recombinant protein with a six-histidine tag was purified to homogeneity by a two-column purification procedure, Ni2+ nitriloacetic acid/agarose followed by carboxymethyl-Sepharose chromatography. Unlike most RTs, the purified MMTV RT is enzymically active as a monomer even after binding a DNA substrate. Like all RTs studied, the recombinant MMTV RT possesses RNA-dependent and DNA-dependent DNA polymerase activities as well as RNase H activity, all of which show a preference for Mg2+ over Mn2+ ions. Other features of these enzymic activities, such as extension of DNA primers, processivity of DNA synthesis, pH dependence, steady-state kinetic constants, effects of Na+ or K+ ions and sensitivity to a thiol-specific reagent and to a zinc chelator, have been evaluated. The catalytic properties of MMTV RT were compared with those of the well-studied RT of HIV-1, the causative agent of AIDS. Interestingly, MMTV RT exhibits a high sensitivity to nucleoside triphosphate analogues (which are known to be potent inhibitors of HIV RTs and are being used as the major anti-AIDS drugs), as high as that of HIV-1 and HIV-2 RTs. Furthermore the recombinant MMTV RT shows a processivity of DNA synthesis higher than that of HIV-1 RT.


Plasma exchange in polyneuropathy associated with monoclonal gammopathy of undetermined significance.
Polyneuropathy associated with monoclonal gammopathy of undetermined significance (MGUS) has been treated with plasma exchange, intravenous immune globulin, and chemotherapy, but the effectiveness of these treatments remains uncertain. We randomly assigned 39 patients with stable or worsening neuropathy and MGUS of the IgG, IgA, or IgM type to receive either plasma exchange twice weekly for three weeks or sham plasma exchange, in a double-blind trial. The patients who initially underwent sham plasma exchange subsequently underwent plasma exchange in an open trial. In the double-blind trial, the average neuropathy disability score improved by 2 points from base line (from 62.5 to 60.5) in the sham-exchange group and by 12 points (from 58.3 to 46.3) in the plasma-exchange group (P = 0.06). A similar difference was observed in the weakness score, a component of the neuropathy disability score (improvement, 1 and 10 points, respectively; P = 0.07). After treatment the summed compound muscle action potentials of motor nerves were 1.2 mV lower (worse) than at base line in the sham-exchange group and 0.4 mV higher (better) in the plasma-exchange group (P = 0.07). The greater degree of improvement with plasma exchange was equal in magnitude to or greater than the difference between not being able to walk on the heels or toes and being able to perform these activities. Changes in the vibratory detection threshold, summed motor-nerve conduction velocity, and sensory-nerve action potentials did not differ significantly between the treatment groups. In the open trial, in which patients who initially underwent sham exchange were treated with plasma exchange, the neuropathy disability score (P = 0.04), weakness score (P = 0.07), and summed compound muscle action potentials (P = 0.07) improved more with plasma exchange than they had with sham exchange. In both the double-blind and the open trial, those with IgG or IgA gammopathy had a better response to plasma exchange than those with IgM gammopathy. Plasma exchange appears to be efficacious in neuropathy associated with MGUS, especially of the IgG or IgA type.


Concentrations of organochlorine pesticides and polychlorinated biphenyls in amphipods (Gammarus lacustris) along an elevation gradient in mountain lakes of western Canada.
Populations of the amphipod Gammarus lacustris were examined for their concentrations of organochlorine pesticides and polychlorinated biphenyls (PCBs) from seven lakes spanning a 1,300-m elevation gradient in Alberta, Canada. The concentrations of several of the semivolatile organochlorine compounds ([SVOCs], vapor pressure > 0.03 Pa at 20 degrees C) increased at higher altitudes. This pattern was generally not observed among the less volatile organochlorines ([LVOCs]. vapor pressure < 0.03 Pa at 20 degrees C). These same SVOC compounds have been previously shown to increase at high latitudes as a result of their long-range transport and preferential deposition in cold climates. We also show that populations of G. lacustris at high elevations have slower growth rates and store more lipids than populations at lower elevations. To resolve the colinearity of independent variables, we used multiple regression to identify patterns of contaminant concentrations in this data set. Multiple regressions showed that the effect of elevation, lipid content, and temperature on contaminant concentrations was no longer significant once the growth rate of Gammarus was included as an independent variable. This study shows that enrichment of SVOCs occurs in Gammarus at high altitudes in Alberta, Canada, and that growth rate (biodilution) appears to be the primary influence. Because Gammarus is an important trophic link in aquatic foodwebs in these environments, enhanced concentrations of toxicants in prey may increase their biomagnification in top predators of high-altitude lakes.


Identification and Synthesis of Selected In Vitro Generated Metabolites of the Novel Selective Androgen Receptor Modulator (SARM) 2f.
Among anabolic agents, selective androgen receptor modulators (SARMs) represent a new class of potential drugs that can exhibit anabolic effects on muscle and bone with reduced side effects due to a tissue-selective mode of action. Besides possible medical applications, SARMs are used as performance-enhancing agents in sports. Therefore, they are prohibited by the World Anti-Doping Agency (WADA) in and out of competition. Since their inclusion into the WADA Prohibited List in 2008, there has been an increase in not only the number of adverse analytical findings, but also the total number of SARMs, making continuous research into SARMs an ongoing topic in the field of doping controls. 4-((2R,3R)-2-Ethyl-3-hydroxy-5-oxopyrrolidin-1-yl)-2-(trifluoromethyl)benzonitrile (SARM 2f) is a novel SARM candidate and is therefore of particular interest for sports drug testing. This study describes the synthesis of SARM 2f using a multi-step approach, followed by full characterization using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance spectroscopy (NMR). To provide the first insights into its biotransformation in humans, SARM 2f was metabolized using human liver microsomes and the microsomal S9 fraction. A total of seven metabolites, including phase I and phase II metabolites, were found, of which three metabolites were chemically synthesized in order to confirm their structure. Those can be employed in testing procedures for routine doping controls, further improving anti-doping efforts.


[Post-Covid osteomyelitis of the facial bones].
The pandemic of coronavirus infection existed from 2019 to 2023. The World Health Organization (WHO) has announced on May 5, 2023 that the pandemic had ended. However, it does not cease to have an adverse effect on the health of the world population. Necrotic lesions of the bones of the facial skeleton are now a characteristic sign of a severe coronavirus infection. We conducted a review of scientific publications that reflected the relationship between coronavirus and necrotic processes of the skull bones, methods of treatment, prevention and the latest developments in this direction. The purpose of this article is to review existing studies on Post-Covid osteomyelitis of facial bones, its impact, features of the clinical picture of this disease, analysis of methods and means of treatment of this group of patients. Analysis of literature data has shown that the search for an ideal dressing material continues, especially the developments of native developers stand emphasized. The advantages of modern materials over traditional ones have become unquestionable, but further research in this direction is required.


Turnover of phosphomonoester groups and compartmentation of polyphosphoinositides in human erythrocytes.
The turnover of phosphomonoester groups of phosphatidylinositol 4-phosphate (PtdIns4P) and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] was investigated in human erythrocytes by short-term labelling with [32P]Pi. The procedure applied ensured a quantitative extraction of erythrocyte polyphosphoinositides as well as their reliable separation for the determinations of pool sizes and specific radioactivities. The pool sizes of phosphatidylinositol (PtdIns), PtdIns4P and PtdIns(4,5)P2 are 25, 11 and 44 nmol/ml of cells respectively. Under steady-state conditions, the phosphorylation fluxes from [gamma-32P]ATP into PtdIns4P and PtdIns(4,5)P2 are in the ranges 14-22 and 46-94 nmol X h-1 X ml of cells-1 respectively. Only 25-60% of total PtdIns4P and 6-10% of total PtdIns(4,5)P2 take part in the rapid tracer exchange, i.e. are compartmentalized. In isolated erythrocyte ghosts, the turnover of PtdIns4P approximately corresponds to that in intact erythrocytes, although any compartmentation can be excluded in this preparation. Under the conditions of incubation employed, the turnover of PtdIns(4,5)P2 is more than one order of magnitude smaller in isolated ghosts than that obtained for intact erythrocytes.


Involvement of organelles and inter-organellar signaling in the pathogenesis of HIV-1 associated neurocognitive disorder and Alzheimer's disease.
Endolysosomes, mitochondria, peroxisomes, endoplasmic reticulum, and plasma membranes are now known to physically and functionally interact with each other. Such findings of inter-organellar signaling and communication has led to a resurgent interest in cell biology and an increased appreciation for the physiological actions and pathological consequences of the dynamic physical and chemical communications occurring between intracellular organelles. Others and we have shown that HIV-1 proteins implicated in the pathogenesis of neuroHIV and that Alzheimer's disease both affects the structure and function of intracellular organelles. Intracellular organelles are highly mobile, and their intracellular distribution almost certainly affects their ability to interact with other organelles and to regulate such important physiological functions as endolysosome acidification, cell motility, and nutrient homeostasis. Indeed, compounds that acidify endolysosomes cause endolysosomes to exhibit a mainly perinuclear pattern while compounds that de-acidify endolysosomes cause these organelles to exhibit a larger profile as well as movement towards plasma membranes. Endolysosome pH might be an early event in the pathogenesis of neuroHIV and Alzheimer's disease and in terms of organellar biology endolysosome changes might be upstream of HIV-1 protein-induced changes to other organelles. Thus, inter-organellar signaling mechanisms might be involved in the pathogenesis of neuroHIV and other neurological disorders, and a better understanding of inter-organellar signaling might lead to improved therapeutic strategies.


The role of molecular genetics in chemoprevention studies of prostate cancer.
Research into the molecular genetics of prostate cancer to date has largely focused on the possible existence of one or several single-locus high-penetrance susceptibility genes and several candidate regions have been Identified, but confirmatory studies of these regions have been inconclusive. Increasingly, attention has turned to identification of candidate genes which may increase prostate cancer risk because their products play an important role in possible etiological pathways for prostate cancer. Of various such pathways which have been suggested for prostate cancer, the best studied in terms of molecular genetics is the androgen signalling pathway. Two genes in this pathway, the androgen receptor (AR) gene and the steroid 5-alpha reductase type II (SRD5A2) gene, have been under particular scrutiny and polymorphic markers in each of these genes that reproducibly predict prostate cancer risk have been identified. Such studies may have important implications for prostate cancer chemoprevention trials. As etiological pathways become better understood at the molecular level, piecing together multiple genetic variants in a pathway will allow identification of high-risk individuals and potential targets for chemopreventive interventions. Moreover, understanding the role of these genes in prostate cancer etiology may help in defining heterogeneity in response to such interventions. Finally, these genes or their products may themselves be legitimate targets for building a chemoprevention strategy.


PRMT4 promotes hepatocellular carcinoma progression by activating AKT/mTOR signaling and indicates poor prognosis.
Background: Protein arginine methyltransferase 4 (PRMT4) has been reported to play a role in several common cancers; however, the function and mechanism of PRMT4 in hepatocellular carcinoma (HCC) are not fully understood. This study aimed to investigate the role and mechanism of PRMT4 in the progression of HCC. Methods: PRMT4 expression and clinicopathological characteristics were investigated using an HCC tissue microarray (TMA) consisting of 140 patient samples analyzed by immunohistochemistry. CCK-8, crystal violet and Transwell assays were used to determine cell proliferation, colony formation, migration, and invasion of HCC cell lines in which PRMT4 was overexpressed or downregulated. The underlying mechanism of PRMT4 function was explored by Western blot assays. Results: PRMT4 was highly expressed in HCC tumor tissues compared to adjacent nontumor tissues. PRMT4 expression was significantly associated with alpha-fetoprotein levels, tumor size, satellite nodules, and microvascular invasion. Patients with higher PRMT4 expression had a shorter survival time and higher recurrence rate. Functional studies demonstrated that PRMT4 overexpression promoted HCC cell proliferation, migration, and invasion in vitro, while knocking down PRMT4 inhibited these malignant behaviors. Additional results revealed that PRMT4 promoted the progression of HCC cells via activation of the AKT/mTOR signaling pathway. Furthermore, inhibition of the AKT/mTOR signaling by MK2206 or rapamycin significantly attenuated PRMT4-mediated malignant phenotypes. Conclusions: This study suggests that PRMT4 may promote the progression of HCC cells by activating the AKT/mTOR signaling pathway, which may be a valuable biomarker and potential target for HCC.


Effects of temperature on plasmid stability and penicillinase productivity in a transformant of Bacillus stearothermophilus.
After transforming host cells of Bacillus stearothermophilus CU21 with a recombinant plasmid pLP11 that harbored constitutive penicillinase genes of B. licheniformis CO1, both the stability of the plasmid and specific rate of penicillinase production were studied. The temperature at which the plasmid could be kept in a stable fashion in the transformant of B. stearothermophilus CU21 (pLP11) ranged nearly from 44 to 50 degrees C, irrespective of batch and continuous cultures. Continuous and steady-state cultures of the transformant could only be realized within this narrower temperature range. Indeed, the approximate temperature ranges of growth for the host and transformant were from 40 to 70 degrees C and from 40 to 63 degrees C, respectively. Clearly, the upper limit for the growth temperature of host cells decreased when they were transformed. Kinetic patterns of penicillinase production in continuous culture of the transformant (with plasmid) from 44 to 50 degrees C differed remarkably from that of B. licheniformis CO1 (without plasmid) at 37 degrees C.


The changing living arrangements of young adults in the UK.
Patterns of leaving the parental home and transitions to independent living, partnership and family formation in the UK are now far more protracted and diverse than they were in the recent past. Over the past twenty years there have been significant changes in the institutional and structural context within which young adults make these transitions. This article examines changes over the past twenty years in the living arrangements of young men and women aged 16 to 34 years and how the proportions living with their parents differ by geographical region, education and economic activity.


Developing an active-learning app to improve critical thinking: item selection and gamification effects.
Critical thinking (CT) is widely recognized as an important skill and attitude in this modern world, but few apps (web-based or installed on devices) have been developed to effectively train it. There is also little research on what kind of content to put into such apps and in what order, if the content is a series of reasoning questions that are intended as CT exercises. Therefore, this research project, consisting of two studies, tries to demonstrate how exercise questions can be presented to learners to sustain their motivation to work on multiple-choice CT questions. In Study 1, question banks were drawn from popular workbooks for CT and verbal reasoning. The questions were ranked in terms of difficulty based on the participation of university students (N = 73). In Study 2, the questions were loaded onto two types of web-based apps: (1) one that sequentially gives multiple-choice questions with immediate feedback and (2) one with minimum gamification of group/individual competition. The experiment to examine the effect of the gamification was conducted (N = 114). Both groups with and without gamification showed improvements in the scores of the pre-/post-tests using comparable questions, but there was no clear effect of gamification. These findings show that an effective CT app can be developed using existing question banks but that the effect of gamification needs further research.


In vivo behaviour of long-circulating liposomes in blood vessels in hamster inflammation and septic shock models-use of intravital fluorescence microscopy.
This study aimed to observe liposome uptake by leukocytes in vivo. The study was performed on skin by using a dorsal skin-fold chamber implanted in golden hamsters using intravital microscopy. 5 and 6-CF-encapsulated polyethylene glycolated liposomes were injected intravenously. The skin microcirculation was observed with an intravital Eclipse E800 Nikon microscope (using x40, x80 magnification) fitted with a Xenon light source and an epifluorescence assembly (excitation, 470 nm, FWHM 40 nm; emission, 540 nm, FWHM 40 nm). An ultra-high sensitivity videocamera mounted on the microscope projected the image onto a monitor, and the images (720 x 576 pixels) were recorded for playback analysis with a digital video cassette recorder. An acute inflammatory response was obtained by removing one complete layer of skin and the underlying fascia and avascular tissue on the opposing side of the flap corresponding to an area equivalent to the window aperture. Using this model and set-up, leukocyte rolling and adhesion were easily observed and the entry of PEGylated liposomes into hamster blood leukocytes was studied for a period of 6 h. PEGylated liposomes were clearly identified alone inside the blood flow and inside the leukocytes as soon as the inflammatory reaction appeared. This study shows for the first time that blood leukocytes in their natural milieu of whole blood are capable of interacting with, and taking up, liposomes. This observation is in accordance with previous in vitro studies.


Simultaneous observation of the lever arm and head explains myosin VI dual function.
Myosin VI is an adenosine triphosphate (ATP)-driven dimeric molecular motor that has dual function as a vesicle transporter and a cytoskeletal anchor. Recently, it was reported that myosin VI generates three types of steps by taking either a distant binding or adjacent binding state (noncanonical hand-over-hand step pathway). The adjacent binding state, in which both heads bind to an actin filament near one another, is unique to myosin VI and therefore may help explain its distinct features. However, detailed information of the adjacent binding state remains unclear. Here simultaneous observations of the head and tail domain during stepping are presented. These observations show that the lever arms tilt forward in the adjacent binding state. Furthermore, it is revealed that either head could take the subsequent step with equal probability from this state. Together with previous results, a comprehensive stepping scheme is proposed; it includes the tail domain motion to explain how myosin VI achieves its dual function.


An extensive cocktail approach for rapid risk assessment of in vitro CYP450 direct reversible inhibition by xenobiotic exposure.
Acute exposure to environmental factors strongly affects the metabolic activity of cytochrome P450 (P450). As a consequence, the risk of interaction could be increased, modifying the clinical outcomes of a medication. Because toxic agents cannot be administered to humans for ethical reasons, in vitro approaches are therefore essential to evaluate their impact on P450 activities. In this work, an extensive cocktail mixture was developed and validated for in vitro P450 inhibition studies using human liver microsomes (HLM). The cocktail comprised eleven P450-specific probe substrates to simultaneously assess the activities of the following isoforms: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 2J2 and subfamily 3A. The high selectivity and sensitivity of the developed UHPLC-MS/MS method were critical for the success of this methodology, whose main advantages are: (i) the use of eleven probe substrates with minimized interactions, (ii) a low HLM concentration, (iii) fast incubation (5min) and (iv) the use of metabolic ratios as microsomal P450 activities markers. This cocktail approach was successfully validated by comparing the obtained IC50 values for model inhibitors with those generated with the conventional single probe methods. Accordingly, reliable inhibition values could be generated 10-fold faster using a 10-fold smaller amount of HLM compared to individual assays. This approach was applied to assess the P450 inhibition potential of widespread insecticides, namely, chlorpyrifos, fenitrothion, methylparathion and profenofos. In all cases, P450 2B6 was the most affected with IC50 values in the nanomolar range. For the first time, mixtures of these four insecticides incubated at low concentrations showed a cumulative inhibitory in vitro effect on P450 2B6.


Adult respiratory distress syndrome: causes, morbidity, and mortality.
The increased microvascular permeability of the adult respiratory distress syndrome (ARDS) results in multiple organ injury with the lungs and subsequently the kidneys as the principal targets. Direct pulmonary injury, for example aspiration of gastric contents, and indirect injury, for instance abdominal sepsis, can lead to the syndrome. The predominant infectious cause of the syndrome is gram-negative aerobic rods. Even in patients with noninfectious causes of the syndrome, the incidence of subsequent pneumonia and sepsis with gram-negative aerobic rods is high so that these organisms contribute greatly to the morbidity and mortality that result from the syndrome. Activation of polymorphonuclear leukocytes and platelets and proteolysis of complement and coagulation components contribute to the pathogenesis of the syndrome, particularly from the indirect or blood-borne causes. Therapy is largely supportive and has not progressed substantially since the introduction of positive end expiratory pressure with mechanical ventilation. The outlook for recovery of lung function of survivors is generally good, but only one in three with the syndrome survives. Because of the inflammatory properties of the syndrome, current interest for therapy centers on parenteral use of nonsteroidal antiinflammatory agents and prostaglandins of the E and I series.


Systematic review and QSPR analysis of chemical penetration through the nail to inform onychomycosis candidate selection.
Recalcitrant nail plate infections can be life-long problems because localizing antifungal agents into infected tissues is problematic. In this systematic review, guided by the SPIDER method, we extracted chemical nail permeation data for 38 compounds from 16 articles, and analyzed the data using quantitative structure-property relationships (QSPRs). Our analysis demonstrated that low-molecular weight was essential for effective nail penetration, with <120 g/mol being preferred. Interestingly, chemical polarity had little effect on nail penetration; therefore, small polar molecules, which effectively penetrate the nail, but not the skin, should be set as the most desirable target chemical property in new post-screen onychomycosis candidate selections.


Solutions to electromagnetic interference problems between cochlear implants and GSM phones.
For persons using cochlear implants, electromagnetic compatibility (EMC) problems may sometimes be an obstacle to using digital cellular telephones. This study aimed at exploring the benefit of three new assistive listening device prototypes that eliminate or diminish EMC problems. Ten experienced cochlear implant users listened in quiet to running speech samples and a sentence test on a landline phone, a digital cellular phone with and without three prototypes. The subjects' performance was assessed using a sentence test, a subjective visual analog scale, and by ranking the best and the poorest listening condition. Compared to the other test conditions, listening to a digital cellular phone alone revealed, on average, the poorest sentence recognition scores (29%) and the poorest results in four different subjective judgments (the amount of disturbances, the clarity of the message, the quality of the sound, overall judgment) with all three implant systems tested. The prototypes generally helped the implantees to recognize speech better on the cellular telephone (by 10-21 percent units, on average). Use of assistive listening devices and further development of EMC of both cochlear implant systems and digital cellular phones needs to take place to enable smooth use of digital cellular phones for all implantees.


The role of RANKL/RANK/OPG system in the canine model of hip periprosthetic infection osteolysis.
We aimed to investigate whether the RANKL/RANK/OPG system is associated with the incidence of periprosthetic osteolysis with septic loosening, and to investigate the differences of RANKL/RANK/OPG system expression in synovial fluid surrounding the normal and septic loosening hip prosthesis in canine models. Twelve healthy adult mongrel canines were divided into two groups: experimental and control. Femoral head and stem replacements were conducted on the right side in both groups. The experimental group received the bacteria fluid intra-articular injection and the other group received the same amount of saline in the same day. The synovial fluid samples were gathered at the 1st, 2nd, 4th, 8th, 12th, 16th and 19th week after the bacteria fluid intra-articular injection for enzyme-linked immunosorbent assay (ELISA), the expression of the RANKL/RANK/OPG system. Surgery on all animals was successful. Two dogs were excluded from the analysis of the result because of a surgery infection or death. The ELISA of the synovial fluid revealed that the ratio of RANKL/OPG showed a significant upward trend (p≤0.05) with time in the test group but the ratio of RANKL/OPG in the control group changed slowly over time (p>0.05). The ratio of RANKL/OPG value between the test and control group showed a significant upward trend, but had no statistical difference (p>0.05) over time. It could be concluded that the RANKL/RANK/OPG system is correlated with the incidence of periprosthetic osteolysis with septic loosening. Consequently, imbalance RANKL/RANK/OPG system was related to periprosthetic osteolysis with septic loosening.


[Risk factors, prevention and therapy of fibrin reactions after IOL implantation].
Postoperative fibrinous reactions following the small-incision technique seem to be very rare. Problem cases can be treated with tissue Plasmin Activator. To investigate the new prophylactic and therapeutic possibilities the last 2,056 cataract procedures with PMMA lens implantation were statistically analyzed. In 152 (7.4%) fibrin cases we observed after ECCE (n = 586) 12.8%, after phacoemulsification (PE) with sutured 6.5-mm corneoscleral incision (n = 546) 7.0% and after PE with clear corneal selfsealing 4.1-mm incision (n = 924) only 3.9% fibrinous reactions (p < 0.01). In case of ECCE the rate of fibrinous reactions amounted to 22.4% following prolonged procedures (< 50 min), while it was low as 9.5% with a short period of time (< 30 min) (p < 0.01). In 8 steroid-resistant cases we applied 25 micrograms Plasmin Activator intraocularly. The injections led to complete dissolution of the fibrinous membranes without complications. Postoperative fibrinous reactions can be reduced statistically significantly by application of the corneal small-incision technique. The most efficient therapy is intraocular Plasmin Activator fibrinolysis.


Chromosomal effects on mutability in the P-M system of hybrid dysgenesis in Drosophila melanogaster.
Two manifestations of hybrid dysgenesis were studied in flies with chromosomes derived from two different P strains. In one set of experiments, the occurrence of recessive X-linked lethal mutations in the germ cells of dysgenic males was monitored. In the other, the behavior of an X-linked P-element insertion mutation, snw, was studied in dysgenic males and also in dysgenic females. The chromosomes of one P strain were more proficient at causing dysgenesis in both sets of experiments. However, there was variation among the chromosomes of each strain in regard to the ability to induce lethals or to destabilize snw. The X chromosome, especially when it came from the stronger P strain, had a pronounced effect on both measures of dysgenesis, but in combination with the major autosomes, these effects were reduced. For the stronger P strain, the autosomes by themselves contributed significantly to the production of X-linked lethals and also had large effects on the behavior of snw, but they did not act additively on these two characters. For this strain, the effects of the autosomes on the X-linked lethal mutation rate suggest that only 1/100 P element transpositions causes a recessive lethal mutation. For the weaker P strain, the autosomes had only slight effects on the behavior of snw and appeared to have negligible effects on the X-linked lethal mutation rate. Combinations of chromosomes from either the strong or the weak P strain affected both aspects of dysgenesis in a nonadditive fashion, suggesting that the P elements on these chromosomes competed with each other for transposase, the P-encoded function that triggers P element activity. Age and sex also influenced the ability of chromosomes and combinations of chromosomes to cause dysgenesis.


Sleep disordered breathing and post-discharge mortality in patients with acute heart failure.
Hospitalizations for heart failure are associated with a high post-discharge risk for mortality. Identification of modifiable predictors of post-discharge mortality during hospitalization may improve outcome. Sleep disordered breathing (SDB) is the most common co-morbidity in heart failure patients. Prospective cohort study of patients hospitalized with acute heart failure (AHF) in a single academic heart hospital. Between January 2007 and December 2010, all patients hospitalized with AHF who have left ventricular ejection fraction (LVEF) ≤ 45% and were not already diagnosed with SDB were the target population. Patients underwent in-hospital attended polygraphy testing for SDB and were followed for a median of 3 years post-discharge. Mortality was recorded using national and state vital statistics databases. During the study period, 1117 hospitalized AHF patients underwent successful sleep testing. Three hundred and forty-four patients (31%) had central sleep apnoea (CSA), 525(47%) patients had obstructive sleep apnoea (OSA), and 248 had no or minimal SDB (nmSDB). Of those, 1096 patients survived to discharge and were included in the mortality analysis. Central sleep apnoea was independently associated with mortality. The multivariable hazard ratio (HR) for time to death for CSA vs. nmSDB was 1.61 (95% CI: 1.1, 2.4, P = 0.02). Obstructive sleep apnoea was also independently associated with mortality with a multivariable HR vs. nmSDB of 1.53 (CI: 1.1, 2.2, P = 0.02). The Cox proportional hazards model adjusted for the following covariates: LVEF, age, BMI, sex, race, creatinine, diabetes, type of cardiomyopathy, coronary artery disease, chronic kidney disease, discharge systolic blood pressure <110, hypertension, discharge medications, initial length of stay, admission sodium, haemoglobin, and BUN. This is the largest study to date to evaluate the effect of SDB on post-discharge mortality in patients with AHF. Newly diagnosed CSA and OSA during AHF hospitalization are independently associated with post-discharge mortality.


Prediction of ischaemic and haemorrhagic stroke by self-measured blood pressure at home: the Ohasama study.
To examine the predictive value of self-measured blood pressure values taken at home (home blood pressure) for risk of stroke and subtypes. We obtained home blood pressure measurements from 1702 people, aged > or =40 years, without a history of stroke, in the general population in Japan, and continued follow-up after a mean period of 10.6 years. The prognostic significance of blood pressure for stroke risk was examined using the Cox proportional hazards regression model, which was adjusted for possible confounding factors. There was a linear relationship between home blood pressure and risk of stroke and subtypes. On average, each 10/5 mmHg elevation in home systolic/diastolic blood pressure respectively, was associated with an approximately 30/20% respectively, higher risk of total stroke. A similar relationship was observed for the risk of haemorrhagic stroke (intracerebral and subarachnoid haemorrhage), and the risk of ischaemic stroke [cerebral infarction and transient ischaemic attack (TIA)]. The risk of stroke and subtypes showed a significantly greater relation with home blood pressure values compared to conventional blood pressure values. This is the first study to demonstrate that home blood pressure is an independent predictor for haemorrhagic and ischaemic stroke, in the general population.


Evaluation of the C-MAC Miller Video Laryngoscope Sizes 0 and 1 During Tracheal Intubation of Infants Less Than 10 kg.
Video laryngoscopy has primarily been developed to assist in difficult airways. Using video laryngoscopy in pediatric airway management is an up-and-coming topic. The aim of the presented study was to compare the intubation conditions obtained when using the C-MAC video laryngoscope with Miller blades sizes 0 and 1 for standard direct laryngoscopy and indirect laryngoscopy in children weighing less than 10 kg. This was a prospective study. The study was performed in a university hospital. Following ethical approval, 86 infants weighing less than 10 kg and undergoing surgery under general anesthesia were studied prospectively. Indirect and direct laryngoscopy either with C-MAC Miller blade size 0 or size 1. First, direct laryngoscopy was performed, and the best obtained view was graded without looking at the video monitor. A second investigator blinded to the view obtained under direct laryngoscopy graded the laryngeal view on the video monitor. Time to intubation, intubation conditions, and intubation attempts were recorded. In infants less than 10 kg, intubation conditions were excellent. There were no significant differences between the use of Miller blade 0 or 1 in reference to Cormack-Lehane grade, time to intubation, time to best view, or intubation attempts. Comparing direct and indirect intubation conditions using either Miller blade 0 or 1 revealed that the use of indirect laryngoscopy provided a significantly better view (P < 0.05) of the vocal cords. In 3 infants weighing more than 8 kg, the Miller blade 0 was described as too short and narrow for intubation. Both devices allowed for an excellent visualization of the vocal cords.


Tyrosine phosphorylation of the triple gene block protein 3 regulates cell-to-cell movement and protein interactions of Potato mop-top virus.
Functions of viral proteins can be regulated through phosphorylation by serine/threonine kinases in plants, but little is known about the involvement of tyrosine kinases in plant virus infection. In this study, TGBp3, one of the three movement proteins encoded by a triple gene block (TGB) of Potato mop-top virus (PMTV), was detected for the first time in PMTV-infected plants and found to be tyrosine phosphorylated. Phosphorylation sites (Tyr(87-89) and Tyr(120)) were located in two amino acid motifs conserved in the TGB-containing, rod-shaped plant viruses. Substitution of these tyrosine residues in both motifs was needed to abolish tyrosine phosphorylation of TGBp3. Substitution of Tyr(87-89) with alanine residues enhanced the interaction between TGBp3 and TGBp2 and inhibited cell-to-cell movement of PMTV. On the other hand, substitution of Tyr(120) with alanine resulted in no alteration in the interaction of TGBp3 with TGBp2, but the mutant virus was not infectious. The results suggest that tyrosine phosphorylation is a mechanism regulating the functions of plant virus movement proteins.


The dissociation between perception and action in the Ebbinghaus illusion: nonillusory effects of pictorial cues on grasp.
According to a recently proposed distinction [1] between vision for perception and vision for action, visually guided movements should be largely immune to the perceptually compelling changes in size produced by pictorial illusions. Tests of this prediction that use the Ebbinghaus illusion have revealed only small effects of the illusion on grasp scaling as compared to its effect on perception [2-4]. Nevertheless, some have argued that the small effect on grasp implies that there is a single representation of size for both perception and action [5]. Recent findings, however, suggest that the 2-D pictorial elements, such as those comprising illusory backgrounds, can sometimes be treated as obstacles and thereby influence the programming of grasp [6]. The arrangement of the 2-D elements commonly used in previous studies examining the Ebbinghaus illusion could therefore give rise to an effect on grasp scaling that is independent of its effect on perceptual judgements, even though the two effects are in the same direction. We present evidence demonstrating that when the gap between the target and the illusion-making elements in the Ebbinghaus illusion is equidistant across different perceptual conditions (Figure 1a), the apparent effect of the illusion on grasp scaling is eliminated.


Assessment of Genetic Parameters and Gene Action Associated with Heterosis for Enhancing Yield Characters in Novel Hybrid Rice Parental Lines.
The technology of hybrid rice utilizing heterosis is an essential requirement for achieving food security. The current study was aimed at assessing the genetic parameters and the gene actions of 15 yield-component traits associated with heterosis, in 9 new parental lines of hybrid rice and their generated hybrids. Five cytoplasmic male sterile (CMS) lines were crossed with four restorer (R) lines using twenty generated line × tester designation hybrid combinations. The results revealed that all the traits were controlled by additive and non-additive gene actions. However, the additive variance was the main component of the total genotypic variance. Assessment of the general combining ability (GCA) detected the best combiners among the genotypes. The hybrid combinations that expressed the highest-positive specific combining ability (SCA) for grain-yield were detected. The correlation between the GCA and SCA was evaluated. The hybrid crosses with high-positive heterosis, due to having a better parent for grain yield, were detected. The principal component analysis (PCA) recorded the first four principal axis displayed Eigenvalues >1 and existing variation cumulative of 83.92% in the genotypes for yield component characteristics. Three-dimensional plots corresponding to the studied traits illustrated that the genotypes Guang8A × Giza181, Quan-9311A × Giza179, II-32A × Giza181, and II-32A × Giza179 are classified as possessing superior grain yield.


Normal outcome of pregnancy with ongoing treatment with natalizumab.
Treatment of multiple sclerosis (MS) with natalizumab during pregnancy is not recommended due to potential risks for the foetus. Despite strong advice accidental pregnancies occur. A 32-year old woman with MS since the age of 26 was treated with natalizumab since January 2008. Treatment was stopped April 2011 due to pregnancy plans, but was restarted following an MS relapse. The patient was thoroughly informed about potential foetal risks, but nevertheless she one year later disclosed that she was pregnant in gestational week 15. Treatment was continued, since the first trimester had passed. The pregnancy course was normal and a healthy daughter was born at full gestational term. This is the second known case where natalizumab treatment continued throughout the whole gestational period.


Evaluation of distillers dried grains with solubles as a feed ingredient for broilers.
Two experiments were conducted to evaluate the use of distillers dried grains with solubles (DDGS) from modern ethanol plants in broiler diets. Experiment 1 was a 2 x 2 factorial experiment with diets containing 2 levels of DDGS (0 and 15%) and 2 diet densities (high and low). The high- and low-density diets were formulated to contain 22% CP and 3,050 kcal MEn/kg and 20% CP and 3,000 kcal MEn/kg, respectively. Eight pens of 6 chicks were fed an experimental diet from 0 to 18 d of age. Weight gain and feed efficiency (gain:feed ratio) of the chicks receiving the high-density diets were (P < 0.05) better than those of chicks fed the low-density diets. However, within the 2 density levels there was no difference in performance of chicks fed diets with 0 or 15% DDGS. In experiment 2, 6 replications of 50 chicks were fed 1 of 4 dietary treatments for 42 d. The diets were formulated to be isocaloric and isonitrogenous and contained 0, 6, 12, or 18% DDGS. There was no significant difference in performance or carcass yield throughout the 42 d experiment except for a depression in BW gain and feed conversion when chicks were fed diets with 18% DDGS in the starter period. These studies indicate that DDGS from modern ethanol plants is an acceptable feed ingredient for broiler diets and can be safely used at 6% in the starter period and 12 to 15% in the grower and finisher periods.


The chromatin remodelling protein LSH/HELLS regulates the amount and distribution of DNA hydroxymethylation in the genome.
Ten-Eleven Translocation (TET) proteins convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) leading to a dynamic epigenetic state of DNA that can influence transcription and chromatin organization. While TET proteins interact with complexes involved in transcriptional repression and activation, the overall understanding of the molecular mechanisms involved in TET-mediated regulation of gene expression still remains limited. Here, we show that TET proteins interact with the chromatin remodelling protein lymphoid-specific helicase (LSH/HELLS) in vivo and in vitro. In mouse embryonic fibroblasts (MEFs) and embryonic stem cells (ESCs) knock out of Lsh leads to a significant reduction of 5-hydroxymethylation amount in the DNA. Whole genome sequencing of 5hmC in wild-type versus Lsh knock-out MEFs and ESCs showed that in absence of Lsh, some regions of the genome gain 5hmC while others lose it, with mild correlation with gene expression changes. We further show that differentially hydroxymethylated regions did not completely overlap with differentially methylated regions indicating that changes in 5hmC distribution upon Lsh knock-out are not a direct consequence of 5mC decrease. Altogether, our results suggest that LSH, which interacts with TET proteins, contributes to the regulation of 5hmC levels and distribution in MEFs and ESCs.


Allocating scarce resources in disasters: emergency department principles.
Decisions about medical resource triage during disasters require a planned structured approach, with foundational elements of goals, ethical principles, concepts of operations for reactive and proactive triage, and decision tools understood by the physicians and staff before an incident. Though emergency physicians are often on the front lines of disaster situations, too often they have not considered how they should modify their decisionmaking or use of resources to allow the "greatest good for the greatest number" to be accomplished. This article reviews key concepts from the disaster literature, providing the emergency physician with a framework of ethical and operational principles on which medical interventions provided may be adjusted according to demand and the resources available. Incidents may require a range of responses from an institution and providers, from conventional (maximal use of usual space, staff, and supplies) to contingency (use of other patient care areas and resources to provide functionally equivalent care) and crisis (adjusting care provided to the resources available when usual care cannot be provided). This continuum is defined and may be helpful when determining the scope of response and assistance necessary in an incident. A range of strategies is reviewed that can be implemented when there is a resource shortfall. The resource and staff requirements of specific incident types (trauma, burn incidents) are briefly considered, providing additional preparedness and decisionmaking tactics to the emergency provider. It is difficult to think about delivering medical care under austere conditions. Preparation and understanding of the decisions required and the objectives, strategies, and tactics available can result in better-informed decisions during an event. In turn, adherence to such a response framework can yield thoughtful stewardship of resources and improved outcomes for a larger number of patients.


Boosting of the enhanced permeability and retention effect with nanocapsules improves the therapeutic effects of cetuximab.
Objective: The introduction of therapeutic antibodies (tAbs) into clinical practice has revolutionized tumor treatment strategies, but their tumor therapy efficiency is still far below expectations because of the rapid degradation and limited tumor accumulation of tAbs. Methods: We developed a nanocapsule-based delivery system to induce the self-augmentation of the enhanced permeability and retention (EPR) effect. This system constantly penetrated across the blood-tumor barrier into the tumor while avoiding the attack of tAbs by the immune system. The biodistribution and therapeutic effect were tested with single dose administration of nanocapsule-tAbs in vivo. Results: The accumulation of Nano(cetuximab) within subcutaneous PC9 tumors was gradually enhanced over 6 days after single dose administration, which was contrary to the biodistribution of native cetuximab. Nano(cetuximab) accumulated in tumor tissues via the EPR effect and released cetuximab. The released cetuximab acted on vascular endothelial cells to destroy the blood-tumor barrier and induce self-augmentation of the EPR effect, which in turn contributed to further tumor accumulation of long-circulating Nano(cetuximab). Compared with single dose administration of native cetuximab, Nano(cetuximab) showed an effective tumor suppressive effect for 3 weeks. Conclusions: The nanocapsule-based delivery system efficiently delivered tAbs to tumor tissues and released them to boost the EPR effect, which facilitated further tumor accumulation of the tAbs. This novel self-augmentation of the EPR effect facilitated by the biological characteristics of tAbs and nanotechnology contributed to the improvement of the therapeutic effect of tAbs, and stimulated new ideas for antibody-based tumor therapy.


Unlocking the Asymmetric Hydrogenation of Tetrasubstituted Acyclic Enones.
Asymmetric hydrogenation (AH) of tetrasubstituted olefins generating two stereocenters is still an open topic. There are only a few reports on the AH of tetrasubstituted olefins with conjugated functional groups, while this process can create useful intermediates for the subsequent elaboration of relevant end products. Most of the tetrasubstituted olefins successfully submitted to AH belong to a small number of functional classes; remarkably, the AH of tetrasubstituted acyclic enones still represents an unsolved challenge. Herein, we disclose a class of air-stable Ir-P,N catalysts, prepared in three steps from commercially available amino alcohols, that can hydrogenate, in minutes, a wide range of electronically and sterically diverse acyclic tetrasubstituted enones (including exocyclic ones) with high yields and high enantioselectivities. The factors responsible for the excellent selectivities were elucidated by combining deuterogenation experiments and theoretical calculations. The calculations indicated that the reduction follows an IrI/IrIII mechanism, in which enantioselectivity is controlled in the first migratory insertion of the hydride to the most electrophilic olefinic Cβ and the formation of the hydrogenated product via reductive elimination takes place prior to the coordination of dihydrogen and the subsequent oxidative addition. The potential of the new catalytic systems is demonstrated by the derivatization of hydrogenation products.


Proton modulation of a Ca(2+)-activated K+ channel from rat skeletal muscle incorporated into planar bilayers.
The effect of pH on the activation of a Ca-activated K+ [K(Ca)] channel from rat skeletal muscle incorporated into planar lipid bilayers was studied. Experiments were done at different intracellular Ca2+ and proton concentrations. Changes in pH modified channel kinetics only from the Ca-sensitive face of the channel. At constant Ca2+ concentration, intracellular acidification induced a decrease in the open probability (Po) and a shift of the channel activation curves toward the right along the voltage axis. The displacement was 23.5 mV per pH unit. This displacement was due to a change in the half saturation voltage (Vo) and not to a change in channel voltage dependence. The shifts in Vo induced by protons appeared to be independent of Ca2+ concentration. The slope of the Hill plot of the open-closed equilibrium vs. pH was close to one, suggesting that a minimum of one proton is involved in the proton-driven channel closing reaction. The change in Po with variations in pH was due to both a decrease in the mean open time (To) and an increase in the mean closed time (Tc). At constant voltage, the mean open time of the channel was a linear function of [Ca2+] and the mean closed time was a linear function of 1/[Ca2+]2. Changes in the internal pH modified the slope, but not the intercept of the linear relations To vs. [Ca2+] and Tc vs. 1/[Ca2+]2. On the basis of these results an economical kinetic model of the effect of pH on this channel is proposed. It is concluded that protons do not affect the open-closed reaction, but rather weaken Ca2+ binding to all the conformational states of the channel. Moreover, competitive models in which Ca2+ and H+ cannot bind to the same open or closed state are inconsistent with the data.


Triclosan - an update.
The discovery in 1998 that triclosan has a site-specific action in the bacterial cell as an inhibitor of NADH- or NADPH-dependent enoyl-acyl carrier protein reductase led to a lively debate in the scientific press. The thesis of this debate was that such a mode of action may allow triclosan to induce resistance and cross-resistance in bacterial cells. The debate last saw review in 2004, and this paper aims at updating our knowledge in this area, given recent research on the topic.


Dead wrong--estrogens, osteoporosis cancer and public policy.
Practicing physicians must constantly decide what is the best treatment for each patient. Their decisions are often influenced by prominent professors and by a climate of opinion created by the press and regulatory agencies. If these are wrong, because of pressure from 'consumer advocates', wrong interpretations, or the risks of litigation, physicians may be forced into making inactive decisions which can cause more harm than the treatment under attack. My attention was drawn to this problem by recent actions of the press and the FDA discouraging the use of estrogens because of a putative risk of endometrial cancer. Even if the danger were real, and I consider the evidence faulty, doctors would have to balance risks against benefits. We have 2,718 deaths each year, constant for ten years, from endometrial cancer and at least 50,000 female deaths because of preventable osteoporotic hip fractures. Recent data establish that the doses of estrogen needed to prevent postmenopausal bone loss are lower than those associated with any cancer. History abounds with similar examples. Semmelweis showed that handwashing prevented deaths from puerperal sepsis but was laughed into a madhouse. More recent examples are the long delays in the use of propranolol, cimetidine, bromcryptine, pulmonary surgery, antihypertensive therapy, poliomyelitis vaccination, penicillin and cardiac surgery. An approach to correction of this serious problem will be presented.


Impaired heart rate response to graded exercise. Prognostic implications of chronotropic incompetence in the Framingham Heart Study.
Previous reports have suggested that an attenuated exercise heart rate response may be associated with coronary heart disease risk and with mortality. These observations may parallel the association between reduced heart rate variability during normal activities and adverse outcome. This investigation was designed to look at the prognostic implications of exercise heart rate response in a population-based sample. In this prospective cohort investigation, 1575 male participants (mean age, 43 years) in the Framingham Offspring Study who were free of coronary heart disease, who were not taking beta-blockers, and who underwent submaximal treadmill exercise testing (Bruce protocol) were studied. Heart rate response was assessed in three ways: (1) failure to achieve 85% of the age-predicted maximum heart rate, which has been the traditional definition of chronotropic incompetence; (2) the actual increase in heart rate from rest to peak exercise; and (3) the ratio of heart rate to metabolic reserve used by stage 2 of exercise ("chronotropic response index"). Proportional hazards analyses were used to evaluate the associations of heart rate responses with all-cause mortality and with coronary heart disease incidence during 7.7 years of follow-up. Failure to achieve target heart rate occurred in 327 (21%) subjects. During follow-up there were 55 deaths (14 caused by coronary heart disease) and 95 cases of incident coronary heart disease. Failure to achieve target heart rate, a smaller increase in heart rate with exercise, and the chronotropic response index were predictive of total mortality and incident coronary heart disease (P <.01). Failure to achieve target heart rate remained predictive of incident coronary heart disease even after adjusting for age, ST-segment response, physical activity, and traditional coronary disease risk factors (adjusted hazard ratio, 1.75; 95% confidence interval, 1.11 to 2.74; P=.02). After adjusting for the same factors, the increase in exercise heart rate remained inversely predictive of total mortality (P=.04) and coronary heart disease incidence (P=.0003). The chronotropic response index also was predictive of total mortality (P=.05) and incident coronary heart disease (P=.001) after adjusting for age and other risk factors. An attenuated heart rate response to exercise, a manifestation of chronotropic incompetence, is predictive of increased mortality and coronary heart disease incidence.


Neuroprotective effects of eugenol against aluminiuminduced toxicity in the rat brain.
Aluminium (Al) is a neurotoxic metal that contributes to the progression of several neurodegenerative diseases. The aim of the present study was to evaluate the protective effect of dietary eugenol supplementation against aluminium (Al)- induced cerebral damage in rats. Male Wistar rats were divided into four groups: normal controls, rats fed a diet containing 6,000 μg g-1 eugenol, rats intoxicated daily with aluminium chloride (84 mg kg-1 body weight) p. o. and fed either a basal diet or a eugenol-containing diet. Daily oral administration of Al for four consecutive weeks to rats significantly reduced brain total antioxidant status (TAS) (11.42±0.31 μmol g-1 tissue, p<0.001) with a subsequent significant enhancement of lipid peroxidation (MDA) (32.55±1.68 nmol g-1 tissue, p<0.002). In addition, Al enhanced brain acetylcholinesterase activity (AChE) (46.22±4.90 U mg-1 protein, p<0.001), tumour necrosis factor alpha (TNF-α) (118.72±11.32 pg mg-1 protein, p<0.001), and caspase 3 (Casp-3) (8.77±1.26 ng mg-1 protein, p<0.001) levels, and in contrast significantly suppressed brain-derived neurotrophic factor (BDNF) (82.74±14.53 pg mg-1 protein, p<0.002) and serotonin (5-HT) (1.54±0.12 ng mg-1 tissue, p<0.01) levels. Furthermore, decreased glial fibrillary acidic protein (GFAP) immunostaining was noticed in the striatum of Al-intoxicated rats, compared with untreated controls. On the other hand, co-administration of dietary eugenol with Al intoxication restored brain BDNF (108.76±2.64 pg mg-1 protein) and 5-HT (2.13±0.27 ng mg-1 tissue) to normal levels, enhanced brain TAS (13.43±0.24 μmol g-1 tissue, p<0.05), with a concomitant significant reduction in TNF-α (69.98±4.74 pg mg-1 protein) and Casp-3 (3.80±0.37 ng mg-1 protein) levels (p<0.001), as well as AChE activity (24.50±3.25 U mg-1 protein, p<0.001), and increased striatal GFAP immunoreactivity, compared with Al-treated rats. Histological findings of brain tissues verified biochemical data. In conclusion, eugenol holds potential as a neuroprotective agent through its hydrophobic, antioxidant, and anti-apoptotic properties, as well as its neurotrophic ability against Al-induced brain toxicity in rats.


Internal benchmarking improves precision and reduces animal requirements for determination of fish bioconcentration factors.
The enactment of new chemical regulations has generated a large need for the measurement of the fish bioconcentration factor (BCF). Past experience shows that the BCF determination lacks precision, requires large numbers of fish, and is costly. A new protocol was tested that shortens the experiment from up to 12 weeks for existing protocols to 2 weeks and reduces the number of fish by a factor of 5, while introducing internal benchmarking for the BCF determination. Rainbow trout were simultaneously exposed to 11 chemicals. The BCFs were quantified using one of the test chemicals, musk xylene, as a benchmark. These were compared with BCFs measured in a parallel experiment based on the OECD 305 guideline. The agreement was <20% for five chemicals and between 20%-25% for two further, while two chemicals lay outside the BCF operating window of the experiment and one was lost due to analytical difficulties. This agreement is better than that observed in a BCF Gold Standard Database. Internal benchmarking allows the improvement of the precision of BCF determination in parallel to large reduction in costs and fish requirements.


[Prevention and treatment of postoperative nausea and vomiting (results of a multicenter study)].
Described in the paper are the results of a multicenter study that was conducted at 11 health-care facilities of the Russian Federation and that was based on an analysis of 623 medical records of adult patients operated on with general anesthesia of different variations. There were 154 (24.7%) males and 469 (75.3%) females, aged 16 to 90. The study denoted the need in and possibility of preventing and arresting the PONV; it also outlined those forms of ondansetron (Zofran) that can be used for the purpose, they are: parenteral and lingual pills and rectal suppositories. An assumption was put forward on the significance of PONV conductive factors and on the most effective forms of ondansetron from among the mentioned above. An understanding, resulting from the study, on that PONV is of the multietiology nature and on that its probability increases in proportion to a compatibility of provoking factors, like sex, surgery technique as well as duration of anesthesia and its variation is of importance. The efficiency of medicamental prevention of PONV by a blocker of HT3-receptors, i.e. by ondansetron (Zofran), is to a great extent predetermined by that if the form and, probably, dose of the drug is in line with a combination of the above factors, which needs further research.


Further description of two patients with biallelic variants in NADSYN1 in association with cardiac and vertebral anomalies.
Congenital nicotinamide adenine dinucleotide (NAD) deficiency disorders are associated with pathogenic variants in the genes NADSYN1, HAAO, and KYNU. These disorders overlap with the anomalies present in vertebral, anal, cardiac, tracheoesophageal, radial and renal, and limb anomalies (VATER/VACTERL) association and often result in premature death. Children who survive typically have developmental delays or intellectual disability. Here, we describe two patients with compound heterozygous variants in NADSYN1 who presented with cardiac and vertebral defects overlapping with the VATER/VACTERL association, although the patients did not satisfy criteria for the diagnosis of VATER/VACTERL due to their lack of limb anomalies and significant renal anomalies. One patient survived into childhood with developmental delays and may represent an expansion of the survival data for NADSYN1-associated NAD deficiency disorders. Interestingly, one patient had hypoplastic left heart syndrome (HLHS) and one had an aortic coarctation and transverse hypoplasia of the aortic arch, suggesting that NADSYN1 sequencing should be performed in children presenting with congenital anomalies related to VATER/VACTERL association and with HLHS and aortic arch abnormalities.


Outcomes of Odontoid Process Fracture Surgery.
Odontoid fractures are found in two age groups. In younger patients, they occur following traffic accidents, falls from a height and during sports. In older patients with poor bone quality, they are usually caused by falls from one's own height. Most fractures are stable and do not require surgical treatment. Unstable, severely displaced fractures with neurological deficits require surgical treatment. The aim of the study was to evaluate the surgical and functional outcomes of treatment of odontoid fractures with a cannulated screw. The study enrolled 20 patients that underwent surgery in the years 2020-2022. The patients were divided into two groups: below 60 (group A) and over 60 years of age (group B). Patients were assessed at one day pre-op, one day post-op, 6 weeks following surgery and 3 months following surgery. Imaging studies were performed to assess the angulation and degree of displacement of bone fragments. After the fracture was surgically fixed with a cannulated screw, bone union and the stability of fracture fixation were assessed. Preoperative and postoperative pain intensity as per a VAS, functional status assessed with the NDI questionnaire, quality of life (EQ-VAS) and neurological status (ASIA scale) were compared. Good functional and surgical results were obtained in both groups of patients. Performance and quality of life improved, while pain intensity decreased in all patients. In 3 patients with post-traumatic paresis, the dysfunctions gradually subsided. No general medical or perioperative complications were observed. Bone union of the fracture was achieved in 40% of patients in group B and in 80% in group A. Achieving union did not determine the functional status of patients. On functional radiographs, all places of fixation were stable. 1. Odontoid fracture fixation with a single cannulated screw is a safe method of treatment. It provided a high union rate and good stability of the fracture. 2. Fracture non-union was noted more often in the group of older patients, but it did not negatively impact quality of life and performance.


Frailty significantly impairs the short term prognosis in elderly patients with heart failure.
Frailty is a condition of elderly characterized by increased vulnerability to stressful events with high risk of adverse outcomes. The purpose of this study was to evaluate the association between frailty and adverse outcomes including death and hospitalization due to heart failure in elderly patients. We included patients aged ≥ 65 years with the diagnosis of heart failure. The clinical and laboratory data, echocardiography and ECGs were recorded. Additionally, the frailty scores of the patients were evaluated according to Canadian Study of Health and Aging. All the patients were divided as frail or non-frail. The groups were compared for their characteristics and the occurrence of clinical outcomes. We included 86 eligible patients. The median follow-up time was four months. The mean age was 75 ± 6.5 years. Of these 86 patients, 17 (19.7%) patients encountered an event (death and/or hospitalization). Nine patients (10.4%) died during follow-up. Thirty patients (34.9%) were considered frail. Among the demographic, clinical and laboratory data, only total protein and albumin levels were found to be lower in frail patients (total protein level: 6.8 ± 0.6 g/dL in non-frails, 6.5 ± 0.9 g/dL in frails, P = 0.05; albumin level: 3.8 ± 0.4 g/dL in non-frails, 3.4 ± 0.6 g/dL in frails, P = 0.001). In multivariate analysis, frailty was found to be strongly associated with clinical outcomes in short term. Being frail in an elderly heart failure patient is associated with death and/or hospitalization due to heart failure in short term. Therefore, frailty score should be evaluated for all elderly heart failure patients as a prognostic marker.


gamma-hydroxy unsaturated nitriles: chelation-controlled conjugate additions.
[reaction--see text] Chelation between gamma-hydroxy unsaturated nitriles and Grignard reagents promotes an otherwise difficult anionic conjugate addition reaction. The intermediate chelate is readily generated by deprotonation with t-BuMgCl followed by the addition of a second Grignard reagent that triggers an intramolecular conjugate addition. Structurally diverse Grignard reagents add with equal efficiency, providing an intermediate anion that stereoselectively alkylates BnBr in an overall addition-alkylation reaction.


Muscarinic receptors in canine colonic circular smooth muscle. I. Coexistence of M2 and M3 subtypes.
The parasympathetic neurotransmitter acetylcholine, acting postsynaptically at the smooth muscle muscarinic receptor, is a principle determinant of colonic motility. In order to elucidate the receptor signal-transduction events responsible for muscarinic receptor-induced contraction of colonic circular smooth muscle, we present here and in the accompanying work studies designed to characterize the muscarinic receptors present in colon and to determine their biochemical coupling. Muscarinic receptor subtypes in canine colonic circular smooth muscle were characterized using radioligand binding techniques. The nonselective muscarinic receptor antagonist radioligand [3H]quinuclidinyl benzilate ([3H]QNB) binds rapidly and reversibly to a single class of saturable sites in colon circular smooth muscle membranes, with an affinity (KD) for the antagonist radioligand of 79.8 +/- 12.6 pM and a density of 123.3 +/- 18.7 fmol/mg of protein. Experiments using membranes prepared from isolated cells purified from the circular smooth muscle layer of canine colon (KD = 102.4 +/- 13.5 pM) confirm the smooth muscle origin of the binding and yield a receptor density of 124,340 receptors/cell. The order of potencies of selective muscarinic receptor antagonists in competition with [3H]QNB for binding to colonic receptors is 4-diphenylacetoxy-N-methylpiperidine methobromide greater than methoctramine greater than AF-DX 116 greater than pirenzepine. Unlike other antagonists tested, pirenzepine competition of [3H]QNB binding is biphasic. The high and low affinities deduced from nonlinear fit of the binding data in colon correlate very well with affinities determined for pirenzepine in mixtures of both submandibular gland (M3) and atrium (M2), indicating the presence of two muscarinic receptor subtypes (82% M2, 18% M3) in colon circular smooth muscle. The muscarinic agonist carbachol binds to both high and low affinity sites in colon, and addition of guanine nucleotide (100 microM GTP gamma S) shifts the agonist competition curve to the right, without eliminating high affinity binding sites. Agonist competition studies with a known ratio of M2 and M3 receptors, obtained by mixing pure M2 and M3 populations, predict the result obtained in colon. cDNA probes specific for each of the muscarinic receptors m1 through m4 were hybridized to colon RNA in a Northern blot analysis. Only m2 and m3 probes hybridized to colon RNA, suggesting the presence of both M2 and M3 receptors. Our data demonstrate that the colon circular smooth muscle contains muscarinic receptors of both the M2 and M3 subtypes, which may be coupled to disparate signal transduction pathways important in the physiological actions of acetylcholine in this tissue.