[Allergic sensitization profile in the immigrant population living in the central region of Spain].
The prevalence of allergic diseases has increased worldwide in the last two decades, particularly in developed countries. Respiratory allergy is determined by genetic heredity, influenced by environmental factors. Migration is a good epidemiological model for assessing the influence of the environment. We present the clinical characteristics of respiratory allergy in immigrants in the central region of Spain. We prospectively collected data on all immigrants referred to the allergy units of 7 different hospitals in Madrid, Cuenca and Ciudad Real in March 2010. Respiratory Allergy was diagnosed using a standard study for allergic diseases. Results in immigrants were compared with data from a similar Spanish population. Sixty-two immigrants and 32 Spanish patients were evaluated (63% female, mean age 28.4 years). Their countries of origin were uniformly distributed among 3 macroareas (North of Africa, Latin America, and Eastern Europe). More than 96% presented rhinitis, although persistent rhinitis was more prevalent amongst Latin Americans (76.9%) than in the Spanish population (48%). No differences were observed in asthma prevalence, although immigrants had higher rates of non-controlled and partially controlled asthma. The mean time of onset of symptoms after immigration was 43 months. Grass pollen was the most relevant allergen with the exception of Arab patients. Sensitization to polcalcin was rare. Otherwise, 44.1% of the Spanish population was sensitized to profilin (only 4.88% among immigrants) although this sensitization did not associate allergy to fruits or other vegetables. In summary, a multicentric study is presented where significant differences have been observed in the sensitization pattern and clinical evolution of respiratory allergy in immigrants and the Spanish population.


Collaborative simulation of soft-tissue deformation for virtual surgery applications.
A key challenge of collaborative surgical simulation is to maintain a high level of state consistency among the distributed users under the limitation of network transmission capacity. In this paper, a framework integrating a scalable deformable model and an extensible communication protocol is proposed to meet this challenge. The parameters of the deformable model are obtained by making reference to the biomechanical properties of human soft tissues. Efficient collaboration is achieved by developing the communication protocol and implementing a series of network management approaches, including service management, computation policies, coupling control, token control and availability mechanism. A prototype has been developed by using the client-server network architecture. Experimental results demonstrate that this framework can support collaborative surgical simulation with acceptable network latencies.


CpG oligodeoxynucleotides and interleukin-12 improve the efficacy of Mycobacterium bovis BCG vaccination in mice challenged with M. tuberculosis.
Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only vaccine approved for prevention of tuberculosis. It has been postulated that serial passage of BCG over the years may have resulted in attenuation of its effectiveness. Because interleukin-12 (IL-12) and oligodeoxynucleotides (ODN) containing cytidine phosphate guanosine (CpG) motifs have been shown to enhance Th1 responses in vivo, they were chosen as adjuvants to increase the effectiveness of BCG vaccination. In this report, mice were vaccinated with BCG with or without IL-12 or CpG ODN and then challenged 6 weeks later via the aerosol route with the Erdman strain of M. tuberculosis. Mice vaccinated with BCG alone showed a 1- to 2-log reduction in bacterial load compared with control mice that did not receive any vaccination prior to M. tuberculosis challenge. Moreover, the bacterial loads of mice vaccinated with BCG plus IL-12 or CpG ODN were a further two- to fivefold lower than those of mice vaccinated with BCG alone. As an immune correlate, the antigen-specific production IFN-gamma and mRNA expression in spleen cells prior to challenge were evaluated. Mice vaccinated with BCG plus IL-12 or CpG ODN showed enhanced production of IFN-gamma compared with mice vaccinated with BCG alone. Finally, granulomas in BCG-vaccinated mice were smaller and more lymphocyte rich than those in unvaccinated mice; however, the addition of IL-12 or CpG ODN to BCG vaccination did not alter granuloma formation or result in added pulmonary damage. These observations support a role for immune adjuvants given with BCG vaccination to enhance its biologic efficacy.


Healthcare resource use and indirect costs associated with migraine in Italy: results from the My Migraine Voice survey.
To evaluate the healthcare resource use (HRU) and cost of lost productivity due to migraine among Italians with ≥4 monthly migraine days (MMDs), with a focus on those with ≥2 prior preventive treatment failures (TFs). Data from Italian participants from the My Migraine Voice survey were used to assess migraine-related HRU and migraine's impact on work productivity and daily activities using the Work Productivity and Activity Impairment questionnaire. The mean, annualized cost of lost productivity was estimated using the Human Capital Approach and extrapolated to employed Italian population with ≥4 MMDs to calculate the overall migraine-related indirect cost burden in Italy. Data of 420 participants, enrolled between September 2017 and February 2018, were analyzed (mean age: 38.5 years, 81.2% women, 37.8% with ≥2 TF). During a 6-month period, 57.6% of participants visited general practitioners (mean visits: 4.5), 31.9% neurologists (mean visits: 2.6), and 26.4% headache specialists (mean visits: 2.8). Overall, 32.0% of participants had ≥1 emergency room visit (mean visits: 2.8) and 15.0% had ≥1 hospitalization (mean visits: 2.9) because of migraine in the past 12 months. Participants who were employed (N = 215) reported 15.5% absenteeism, 45.3% presenteeism, 53.8% overall work impairment, and 52.6% activity impairment. The mean annualized indirect cost was estimated to be €14,368. The annual indirect cost burden was estimated to be €7.6 billion for the employed Italian population with ≥4 MMDs. The impact of migraine was particularly high among the ≥2 TF subgroups on all parameters. The indirect cost was estimated to be €15,881 (€5,007 attributed to absenteeism). Migraine-related HRU and indirect costs are high among individuals with ≥4 MMDs (particularly those with ≥2 TF). There is a need for more effective treatments and better management of migraines to reduce the functional and economic burden among this difficult-to-treat population.


Gellan gum-based hydrogels for intervertebral disc tissue-engineering applications.
Intervertebral disc (IVD) degeneration is a challenging clinical problem that urgently demands viable nucleus pulposus (NP) implant materials. The best suited biomaterial for NP regeneration has yet to be identified, but it is believed that biodegradable hydrogel-based materials are promising candidates. In this work, we have developed ionic- and photo-crosslinked methacrylated gellan gum (GG-MA) hydrogels to be used in acellular and cellular tissue-engineering strategies for the regeneration of IVDs. The physicochemical properties of the developed hydrogels were investigated by Fourier-transform infrared spectroscopy, (1) H nuclear magnetic resonance and differential scanning calorimetry. The swelling ability and degradation rate of hydrogels were also analysed in phosphate-buffered saline solution at physiological pH for a period of 30 days. Additionally, the morphology and mechanical properties of the hydrogels were assessed under a scanning electron microscope and dynamic compression, respectively. An in vitro study was carried out to screen possible cytotoxicity of the gellan gum-based hydrogels by culturing rat lung fibroblasts (L929 cells) with hydrogel leachables up to 7 days. The results demonstrated that gellan gum was successfully methacrylated. We observed that the produced GG-MA hydrogels possess improved mechanical properties and lower water uptake ability and degradation rate as compared to gellan gum. This work also revealed that GG-MA hydrogels are non-cytotoxic in vitro, thus being promising biomaterials to be used in IVD tissue-engineering strategies.


Hexose-independent activation of glycogen synthase and pyruvate dehydrogenase by insulin is dissociated in the mouse BC3H-1 cell line.
We have studied the effects of insulin on several aspects of cell metabolism in the insulin-sensitive, nonfusing muscle cell line BC3H-1. In the absence of exogenous hexose, insulin did not alter basal glycogen synthase percentage I activity, or attenuate the increase in intracellular cAMP content, the activation of glycogen phosphorylase a, or the decrease in glycogen synthase I brought about by beta-adrenergic receptor activation with epinephrine. In contrast, both insulin and the tumor-promoting phorbol ester, tetradecanoylyl phorbol acetate markedly increased mitochondrial pyruvate dehydrogenase activity in the absence of hexose. Both glycogen synthase phosphatase and glycogen synthase kinase activities were present in BC3H-1 cell extracts and were regulated in the expected manner by glucose 6-phosphate and cAMP, respectively. Since the pattern of partial insulin resistance seen in BC3H-1 myocytes would require that several potentially insulin-sensitive enzymes be insensitive to insulin-generated signals, the most likely explanation for these data is that the myocytes are defective in some mechanism of insulin signaling which is independent of the mechanism for pyruvate dehydrogenase activation.


[Subtotal duodenopancreatectomy. A new surgical technic for cancer of the pancreas head].
Since May 1980 total duodenopancreatectomy for pancreatic cancer, which caused an operative mortality of 25% in former years, was replaced by a subtotal duodenopancreatectomy. This new technique is discussed in detail. Altogether 30 patients were operated since then without postoperative mortality. The procedure is tolerated by the patients much better than a total pancreatectomy. Only 40% developed diabetes postoperatively. At present 4/24 patients with ductal carcinoma of the cephalic pancreas are alive longer than 20 months. It seems, that subtotal duodenopancreatectomy is as an effective radical operation as total pancreatectomy, with a much lower operative risk for the patient.


Dating of Acute and Subacute Subdural Haemorrhage: A Histo-Pathological Study.
Microscopic study of the organization of the Subdural Haemorrhage (SDH) verified against the time period can help us in the determination of its age which has serious medico-legal implications. Very few studies concerning the dating of SDH are present in the literature. This study was conducted for dating the early subdural haemorrhage by routine histopathological stains. A prospective analytical study was conducted during July 2009 to December 2010. A total of 100 cases (50 males and 50 females) fulfilling the inclusion and exclusion criteria were included in this study. Routine histopathological staining of the subdural haematoma was done. Correlation between the frequency of a given histomorphological phenomenon and the length of the Post-Traumatic Interval (PTI) was evidential. All the histomorphological features, when correlated with PTI groups, were found to be statistically significant, except for Polymorphonuclear Leukocytes (PMN). We concluded that routine histopathology was reliable in the dating of early subdural haemorrhages.


[The affinity of m-cholinomimetics for the m-cholinoreceptors in different tissues].
The affinities of some cholinomimetics (acetylcholine, methacholine, carbachol, oxotremorine, aceclidine and pilocarpine) for muscarinic acetylcholine receptors of various isolated tissues (rat spontaneously beating atria, rat electrically stimulated left atrium, segment of rat aorta precontracted by PGF2 alpha, rat trachea and guinea pig trachea) was estimated in functional experiments. It was established that each of full agonists (acetylcholine, methacholine, carbachol, oxotremorine and aceclidine) has lower affinities for muscarinic M2 receptors of rat atria as compared with its affinity for muscarinic M3 receptors of rat aorta. The partial agonist pilocarpine has similar affinities for muscarinic receptors of rat atria and rat aorta. It was shown that contractile response of guinea pig tracheal smooth muscle is two-staged. It was found that rat and guinea pig tracheal smooth muscle contractions are mediated primarily by muscarinic M3 cholinoceptors and partially by M2 cholinoceptors.


Pentose phosphate pathway flux analysis for glycopeptide antibiotic vancomycin production during glucose-limited cultivation of Amycolatopsis orientalis.
In vivo pentose phosphate pathway (PPP) enzymes such as glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), and transaldolase (TAL) activities as well as ATP- and ADP-level variations of Amycolatopsis orientalis were investigated with respect to glucose concentration and incubation period. G6PDH, 6PGDH, and TAL activities of A. orientalis reached maximum levels at 48 hr for all glucose concentrations used, after which the levels began to decline. G6PDH, 6PGDH, and TAL activities showed positive correlation with the glucose concentration up to 15 g/L, while further increases had an opposite effect. Intracellular ATP level showed a positive correlation with glucose concentrations, while ADP level increased up to 15 g/L. ATP concentration of A. orientalis increased rapidly at 48 hr of incubation, as was the case also for G6PDH, 6PGDH, and TAL activities, although the incubation period corresponding to maximum values of ADP shifted to 60 hr. Production of the glycopeptide antibiotic vancomycin increased with the increases in glucose concentrations up to 15 g/L, by showing coherence in the rates of oxidative and nonoxidative parts of the PPP.


Nuclear quantum effects in water exchange around lithium and fluoride ions.
We employ classical and ring polymer molecular dynamics simulations to study the effect of nuclear quantum fluctuations on the structure and the water exchange dynamics of aqueous solutions of lithium and fluoride ions. While we obtain reasonably good agreement with experimental data for solutions of lithium by augmenting the Coulombic interactions between the ion and the water molecules with a standard Lennard-Jones ion-oxygen potential, the same is not true for solutions of fluoride, for which we find that a potential with a softer repulsive wall gives much better agreement. A small degree of destabilization of the first hydration shell is found in quantum simulations of both ions when compared with classical simulations, with the shell becoming less sharply defined and the mean residence time of the water molecules in the shell decreasing. In line with these modest differences, we find that the mechanisms of the exchange processes are unaffected by quantization, so a classical description of these reactions gives qualitatively correct and quantitatively reasonable results. We also find that the quantum effects in solutions of lithium are larger than in solutions of fluoride. This is partly due to the stronger interaction of lithium with water molecules, partly due to the lighter mass of lithium and partly due to competing quantum effects in the hydration of fluoride, which are absent in the hydration of lithium.


Changes in organization of the endoplasmic reticulum during Xenopus oocyte maturation and activation.
The organization of the endoplasmic reticulum (ER) in the cortex of Xenopus oocytes was investigated during maturation and activation using a green fluorescent protein chimera, immunofluorescence, and electron microscopy. Dense clusters of ER developed on the vegetal side (the side opposite the meiotic spindle) during maturation. Small clusters appeared transiently at the time of nuclear envelope breakdown, disappeared at the time of first polar body formation, and then reappeared as larger clusters in mature eggs. The appearance of the large ER clusters was correlated with an increase in releasability of Ca(2+) by IP(3). The clusters dispersed during the Ca(2+) wave at activation. Possible relationships of ER structure and Ca(2+) regulation are discussed.


Comparative study of droplet-digital PCR and absolute Q digital PCR for ctDNA detection in early-stage breast cancer patients.
Analysis of circulating tumor DNA (ctDNA) is increasingly used for clinical decision-making in oncology. However, ctDNA could represent ≤ 0.1 % of cell-free DNA in early-stage tumors and its detection requires high-sensitive techniques such as digital PCR (dPCR). In 46 samples from patients with early-stage breast cancer, we compared two leading dPCR assays for ctDNA analysis: QX200 droplet digital PCR (ddPCR) system from Bio-Rad which is the gold-standard in the field, and Absolute Q plate-based digital PCR (pdPCR) system from Thermo Fisher Scientific which has not been reported before. We analyzed 5 mL of baseline plasma samples prior to any treatment. Both systems displayed a comparable sensitivity with no significant differences observed in mutant allele frequency. In fact, ddPCR and pdPCR possessed a concordance > 90 % in ctDNA positivity. Nevertheless, ddPCR exhibited higher variability and a longer workflow. Finally, we explored the association between ctDNA levels and clinicopathological features. Significantly higher ctDNA levels were present in patients with a Ki67 score > 20 % or with estrogen receptor-negative or triple-negative breast cancer subtypes. Both ddPCR and pdPCR may constitute sensitive and reliable tools for ctDNA analysis with an adequate agreement in early-stage breast cancer patients.


A comparative analysis of the statistical properties of large mobile phone calling networks.
Mobile phone calling is one of the most widely used communication methods in modern society. The records of calls among mobile phone users provide us a valuable proxy for the understanding of human communication patterns embedded in social networks. Mobile phone users call each other forming a directed calling network. If only reciprocal calls are considered, we obtain an undirected mutual calling network. The preferential communication behavior between two connected users can be statistically tested and it results in two Bonferroni networks with statistically validated edges. We perform a comparative analysis of the statistical properties of these four networks, which are constructed from the calling records of more than nine million individuals in Shanghai over a period of 110 days. We find that these networks share many common structural properties and also exhibit idiosyncratic features when compared with previously studied large mobile calling networks. The empirical findings provide us an intriguing picture of a representative large social network that might shed new lights on the modelling of large social networks.


Laparoscopic totally extraperitoneal (TEP) hernioplasty using two trocars: anatomical landmarks and surgical technique.
Among endoscopic hernioplasties, totally extraperitoneal (TEP) and transabdominal preperitoneal (TAPP) approach are widely accepted alternatives to open surgery, both providing less postoperative pain, hospital length of stay and early return to work. Classical TEP technique requires three skin incisions for placement of three trocars in the midline or in triangulation. To describe a technique using only two trocars for laparoscopic total extraperitoneal for inguinal hernia repair. Extraperitoneal access: place two regular trocars on the midline. The 10 mm is inserted into the subcutaneous in horizontal direction after a transverse infra-umbilical incision and then elevated at 60º angle. The 5 mm trocar is inserted at the same level of the pubis with direct vision. Preperitoneal space dissection: introduction 0º optical laparoscope through the infra-umbilical incision for visualization and preperitoneal dissection; insufflation pressure must be below 12 mmHg. Dissection of some anatomical landmarks: pubic bone, arcuate line and inferior epigastric vessels. Exposure of "triangle of pain" and "triangle of doom". Insertion through the 10 mm trocar polypropylene mesh of 10x15 cm to cover the hernia sites. Peritoneal sac and the dorsal edge of the mesh are repositioned in order to avoid bending or mesh displacement. It is also important to remember that the drainage is not necessary. The 2-port TEP required less financial costs than usual because it is not necessary an auxiliary surgeon to perform the technique. Trocars, suturing material and wound dressing were spared in comparison to the classical technique. Besides, there were only two incisions, which provides a better plastic result and less postoperative pain. The TEP technique using two trocars is an alternative technique which improves cosmetic and financial outcomes.


The effect of tumor burden on ornithine decarboxylase activity in mice.
Polyamines are essential for cell growth of normal and neoplastic tissue, alpha-Difluoromethylornithine (DFMO) is a known irreversible inhibitor or ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis. The purpose of this study was to examine the effects of tumor burden on ODC in tissues of tumor-bearing compared with tumor-free mice. Twenty-eight male Balb/c mice were divided into four groups of 7 each. Groups 1 and 2 were inoculated subcutaneously with 10 x 10(6) MC-26 mouse colon adenocarcinoma cells. Groups 3 and 4 were kept as tumor-free controls. Ten days after inoculation, groups 2 and 4 were injected with DFMO (200 mg/kg) intraperitoneally (IP) while Groups 1 and 3 received saline. Two hours after the injection of DFMO the animals were sacrificed. The tumor, pancreas, kidney, and liver were excised and analyzed for ODC activity. DFMO caused a significant reduction (compared with controls that did not receive DFMO) in the ODC activity of tumors; however, ODC activity of the kidney, pancreas, and liver of tumor-bearing mice was not affected. Additionally, the basal ODC activity in the kidney, liver, and pancreas of tumor-bearing mice was significantly lower compared with tumor-free controls. DFMO lowered ODC activity in the kidney, pancreas, and liver of tumor-free mice. These results suggest that the presence of MC-26 tumor causes systemic effects that alter ODC activity and the response to a known inhibitor of ODC.


Unpacking the enabling factors for hand, cord and birth-surface hygiene in Zanzibar maternity units.
Recent national surveys in The United Republic of Tanzania have revealed poor standards of hygiene at birth in facilities. As more women opt for institutional delivery, improving basic hygiene becomes an essential part of preventative strategies for reducing puerperal and newborn sepsis. Our collaborative research in Zanzibar provides an in-depth picture of the state of hygiene on maternity wards to inform action. Hygiene was assessed in 2014 across all 37 facilities with a maternity unit in Zanzibar. We used a mixed methods approach, including structured and semi-structured interviews, and environmental microbiology. Data were analysed according to the WHO 'cleans' framework, focusing on the fundamental practices for prevention of newborn and maternal sepsis. For each 'clean' we explored the following enabling factors: knowledge, infrastructure (including equipment), staffing levels and policies. Composite indices were constructed for the enabling factors of the 'cleans' from the quantitative data: clean hands, cord cutting, and birth surface. Results from the qualitative tools were used to complement this information.Only 49% of facilities had the 'infrastructural' requirements to enable 'clean hands', with the availability of constant running water particularly lacking. Less than half (46%) of facilities met the 'knowledge' requirements for ensuring a 'clean delivery surface'; six out of seven facilities had birthing surfaces that tested positive for multiple potential pathogens. Almost two thirds of facilities met the 'infrastructure (equipment) requirement' for 'clean cord'; however, disposable cord clamps being frequently out of stock, often resulted in the use of non-sterile thread made of fabric. This mixed methods approach, and the analytical framework based on the WHO 'cleans' and the enabling factors, yielded practical information of direct relevance to action at local and ministerial levels. The same approach could be applied to collect and analyse data on infection prevention from maternity units in other contexts.


Safety, effectiveness and tolerability of sublingual ketamine in depression and anxiety: A retrospective study of off-label, at-home use.
Intravenous and intranasal ketamine have been shown to be effective therapeutic options in patients suffering from treatment-resistant depression (TRD). The use of sublingual (SL), rapid dissolve ketamine tablets (RDT) offers a novel approach for delivery for mental health indications. This study assessed the effectiveness and safety of self-administration of off-label, SL, rapid dissolve ketamine tablets (RDT) at-home for depression and anxiety. Intake scores on the Generalized Anxiety Disorder Screener (GAD-7) and Patient Health Questionnaire (PHQ-9) were compared to scores after treatments of three doses of ketamine RDT, and after six doses of ketamine RDT. After three doses of SL ketamine, 47.6% of patients showed a significant decrease in PHQ-9 scores, and 47.6% of patients showed a significant reduction in GAD-7 scores. Reduction rates were higher in those patients who completed a clinically recommended six doses of RDT ketamine. This study demonstrates that SL ketamine is a novel, safe, and effective treatment for TRD and treatment-resistant anxiety. SL ketamine offers an alternative therapeutic approach to IV ketamine when treating those with TRD.


Farnesoid X receptor knockdown provides significant growth inhibition in hepatocellular carcinoma cells while it does not interfere with the proliferation of primary human hepatocyte-derived cells.
Identification of substances with specific toxicity for carcinoma cells promises to facilitate the development of cancer chemotherapeutics that cause minimal side effects. Here, we show that knockdown of the farnesoid X receptor (FXR) effectively suppresses the proliferation of human hepatocellular carcinoma cell lines HepG2 and HLE accompanied by elevated expression of cyclin-dependent kinase (CDK) inhibitor p16/INK4a and p21/Cip1 proteins. On the other hand, the growth of the primary human hepatocyte-derived cell line Fa2N-4 is not affected by the treatment with FXR siRNA irrespective of marked increases in the mRNAs of p16/INK4a and p21/Cip1. Surprisingly, the expression levels of p16/INK and p21/Cip1 proteins are left unchanged in Fa2N-4 cells that are subjected to the FXR siRNA treatment. Since the expression levels of these CDK inhibitor proteins in FXR-knockdown Fa2N-4 cells were elevated in the presence of proteasomal inhibitor MG132, these CDK inhibitors may be subjected to the proteasomal degradation, thereby counteracting the increased expression of their cognate mRNAs, therefore similar levels of p16 and p21 proteins were observed in control and FXR-knockdown Fa2N-4 cells. These results suggest that FXR-knockdown is effective for inhibiting the proliferation of hepatocellular carcinoma cells, not interfering with the regulatory mechanism of normal hepatocyte growth.


Neurological syndromes and potential triggers associated with antibodies to neuronal surface antigens.
Autoantibodies against surface neuronal antigens have been associated with specific neurological presentations including autoimmune encephalitis (AE), with variable association with neoplasia and infections. We described the phenotype and environmental associations of patients with neurological syndromes associated with antibodies against neuronal surface antigens who were referred to a tertiary center in the South of Brazil. All patients were tested for neuronal autoantibodies using cell-based assays. Clinical, radiological, and laboratory findings were retrospectively reviewed. We identified 16 patients, 15 had subacute, and one had a progressive disease course. Among patients with subacute onset, 11 (73 %) were N-Methyl-d-Aspartate receptor (NMDAr-IgG)+, 3 (20 %) were Leucine-rich Glioma-Inactivated-1 (LGI1-IgG)+, and 1 (6 %) was positive for Glycine receptor-IgG. The patient with a progressive disease course had antibodies against IgLON5. Most patients had disease onset in spring and summer suggesting environmental factors for the development of AE. Also, we observed a different pattern of brain lesions when NMDAr-IgG encephalitis followed herpes encephalitis and a previously unreported association with Rosai-Dorfman-Destombe disease. All patients with encephalopathy met criteria for possible AE and all proven NMDAr-IgG+ met criteria for NMDAr-IgG encephalitis. However, only one LGI1-IgG+ patient fulfilled clinical criteria for limbic encephalitis. All but one received high-dose intravenous methylprednisolone, 11 also had intravenous human immunoglobulin, and 4 plasma exchange. Furthermore, all patients received second-line immunotherapy. Importantly, most patients improved with immunotherapy, even when initiated later in the disease course. We identified seasonal variability associated with neuronal surface antibodies suggesting environmental triggers. Also, we described the coexistence of NMDAr-IgG encephalitis with histiocytosis. In our series, most patients received second-line immunotherapy. We observed neurologic improvement after treatment even in cases of delayed diagnosis. Increasing the recognition and availability of tests and treatments for these conditions is of paramount importance in low- and middle-income countries.


Tracking the recruitment and evolution of snake toxins using the evolutionary context provided by the Bothrops jararaca genome.
Venom is a key adaptive innovation in snakes, and how nonvenom genes were co-opted to become part of the toxin arsenal is a significant evolutionary question. While this process has been investigated through the phylogenetic reconstruction of toxin sequences, evidence provided by the genomic context of toxin genes remains less explored. To investigate the process of toxin recruitment, we sequenced the genome of Bothrops jararaca, a clinically relevant pitviper. In addition to producing a road map with canonical structures of genes encoding 12 toxin families, we inferred most of the ancestral genes for their loci. We found evidence that 1) snake venom metalloproteinases (SVMPs) and phospholipases A2 (PLA2) have expanded in genomic proximity to their nonvenomous ancestors; 2) serine proteinases arose by co-opting a local gene that also gave rise to lizard gilatoxins and then expanded; 3) the bradykinin-potentiating peptides originated from a C-type natriuretic peptide gene backbone; and 4) VEGF-F was co-opted from a PGF-like gene and not from VEGF-A. We evaluated two scenarios for the original recruitment of nontoxin genes for snake venom: 1) in locus ancestral gene duplication and 2) in locus ancestral gene direct co-option. The first explains the origins of two important toxins (SVMP and PLA2), while the second explains the emergence of a greater number of venom components. Overall, our results support the idea of a locally assembled venom arsenal in which the most clinically relevant toxin families expanded through posterior gene duplications, regardless of whether they originated by duplication or gene co-option.


Childhood intelligence, educational attainment and adult body mass index: findings from a prospective cohort and within sibling-pairs analysis.
The mechanisms underlying the observed association of childhood intelligence with body mass index (BMI) are unclear and few studies of this association have been prospective in design. Prospective study in a birth cohort of 5467 individuals who were born in Aberdeen, Scotland between 1950 and 1956 and who responded to a follow-up survey in 2001. Comparison of associations within sibling pairs of the same family to associations between different families in 643 sibling pairs (1286 individuals) who are participants in the main cohort. Childhood intelligence (age 7 years) and educational attainment were both inversely associated with adult BMI (mean age 48 years): the sex- and age-adjusted mean change in adult BMI per s.d. of intelligence was -0.35 kg/m(2) (95% CI: -0.49, -0.21 kg/m(2)) and per unit increase in educational category (seven categories) was -0.28 kg/m(2) (95% CI: -0.34, -0.22). On adjustment for education the association between childhood intelligence and adult BMI attenuated to the null (-0.03 kg/m(2) (-0.19, 0.13 kg/m(2))); other potential confounding or mediating factors had little or only modest effects on this association. The association between education and adult BMI was not affected by adjustment for childhood intelligence or other potential covariates. The within sibling-pair effect of education on adult BMI (-0.06 kg/m(2) (95% CI: -0.26, 0.14)) was weaker than the effect between different families (-0.37 kg/m(2) (95%CI: -0.58, -0.17)), P-value for difference of within sibling and between family effect=0.03. The association of childhood intelligence with adult BMI is attenuated to the null on adjustment for educational attainment, whereas the association of educational attainment with adult BMI appears to be independent of childhood intelligence and other measured covariates. However, our family analyses suggest that fixed family and neighbourhood factors, which are closely matched in siblings of a similar age, explain much of the association between greater educational attainment and lower adult BMI.


Glycyl-tRNA synthetase uses a negatively charged pit for specific recognition and activation of glycine.
The crystal structures of glycyl-tRNA synthetase (GlyRS) from Thermus thermophilus, a homodimeric class II enzyme, were determined in the enzyme-substrate and enzyme-product states corresponding to the first step of aminoacylation. GlyRS was cocrystallized with glycine and ATP, which were transformed by the enzyme into glycyl-adenylate and thus gave the enzyme-product complex. To trap the enzyme-substrate complex, the enzyme was combined with the glycine analog ethanolamine and ATP. The ligands are bound in fixed orientations in the substrate-binding pocket of the N-terminal active site domain, which contains the classical class II aminoacyl-tRNA synthetase (aaRS) fold. Since glycine does not possess a side-chain, much of the specificity of the enzyme is directed toward excluding any additional atoms beyond the alpha-carbon atom. Several carboxylate residues of GlyRS line the glycine binding pocket; two of them interact directly with the alpha-ammonium group. In addition, the enzyme utilizes the acidic character of the pro-L alpha-hydrogen atom by contacting it via a glutamate carboxylic oxygen atom. A guanidino eta-nitrogen atom of the class II aaRS-conserved motif 2 arginine interacts with the substrate carbonyl oxygen atom. These features serve to attract the small amino acid substrate into the active site and to position it in the correct orientation. GlyRS uses class II-conserved residues to interact with the ATP and the adenosine-phosphate moiety of glycyl-adenylate. On the basis of this similarity, we propose that GlyRS utilizes the same general mechanism as that employed by other class II aminoacyl-tRNA synthetases.


Influenza A virus-induced polymorphonuclear leukocyte dysfunction in the pathogenesis of experimental pneumococcal otitis media.
The role of influenza A virus-induced polymorphonuclear leukocyte and eustachian tube dysfunction in the pathogenesis of acute purulent otitis media was studied in chinchillas. Polymorphonuclear leukocyte function, middle ear pressure, and the incidence of pneumococcal otitis media were observed after intranasal inoculation with influenza A virus, Streptococcus pneumoniae, or both. Results showed that depressed negative middle ear pressure and polymorphonuclear leukocyte chemiluminescence and chemotactic activity occurred after influenza inoculation, but not after inoculation with pneumococcus alone. The greatest incidence of pneumococcal otitis media occurred when the pneumococcus was inoculated just before the time of influenza-induced polymorphonuclear leukocyte dysfunction and negative middle ear pressure. Animals that had unilateral tympanostomy tubes placed before inoculation of influenza with pneumococcus showed no difference in the occurrence of pneumococcal otitis media in ventilated and nonventilated ears, suggesting that polymorphonuclear leukocyte dysfunction contributes more to the pathogenesis of pneumococcal otitis media than does negative middle ear pressure in this animal model.


Is Adult Hippocampal Neurogenesis Really Relevant for the Treatment of Psychiatric Disorders?
Adult neurogenesis consists in the generation of newborn neurons from neural stem cells taking place in the adult brain. In mammals, this process is limited to very few areas of the brain, and one of these neurogenic niches is the subgranular layer of the dentate gyrus (DG) of the hippocampus. Adult newborn neurons are generated from quiescent neural progenitors (QNPs), which differentiate through different steps into mature granule cells (GCs), to be finally integrated into the existing hippocampal circuitry. In animal models, adult hippocampal neurogenesis (AHN) is relevant for pattern discrimination, cognitive flexibility, emotional processing and resilience to stressful situations. Imaging techniques allow to visualize newborn neurons within the hippocampus through all their stages of development and differentiation. In humans, the evidence of AHN is more challenging, and, based on recent findings, it persists through adulthood, even if it declines with age. Whether this process has an important role in human brain function and how it integrates into the existing hippocampal circuitry is still a matter of exciting debate. Importantly, AHN deficiency has been proposed to be relevant in many psychiatric disorders, including mood disorders, anxiety, post-traumatic stress disorder and schizophrenia. This review aims to investigate how AHN is altered in different psychiatric conditions and how pharmacological treatments can rescue this process. In fact, many psychoactive drugs, such as antidepressants, mood stabilizers and atypical antipsychotics (AAPs), can boost AHN with different results. In addition, some non-pharmacological approaches are discussed, as well.


An overload hypothesis for pulmonary clearance of UICC amosite fibres inhaled by rats.
Two types of experiments were carried out to examine the effects of deposition and clearance on the accumulation in the lungs of rats of inhaled fibres of UICC amosite. In the first experiment the mass lung burdens of the dust in question were measured as a function of the time at which animals were killed after the cessation of the six week exposure period, and in the second the masses were measured for rats removed from exposure and killed at intervals during the exposure period itself. The experimental conditions were chosen to complement those of earlier work. Taken together with the results of that earlier work, the new results provide the basis for a simple mathematical model of the kinetics of deposition and clearance which appears to account for the major observed trends. Most significantly, there is strong evidence for an overload of clearance at high lung burdens (exceeding about 1500 micrograms/rat), in which a breakdown occurs of the intermediate rate clearance mechanisms (time constants of the order of 12 days). This hypothesis is supported for inhaled asbestos dust, quartz dust, and diesel fume by results obtained elsewhere. Biological explanations for the clearance overload hypothesis are at present speculative, involving discussion of the role of the macrophage in pulmonary clearance. It is believed that the clearance overload hypothesis could have possible consequences for people occupationally exposed to airborne dusts.


Detection and characterization of heterogeneous vancomycin-intermediate Staphylococcus aureus isolates in Canada: results from the Canadian Nosocomial Infection Surveillance Program, 1995-2006.
We describe the epidemiology of heterogeneously resistant Staphylococcus aureus (hVISA) identified in Canadian hospitals between 1995 and 2006. hVISA isolates were confirmed by the population analysis profiling-area under the curve method. Only 25 hVISA isolates (1.3% of all isolates) were detected. hVISA isolates were more likely to have been health care associated (odds ratio [OR], 5.1; 95% confidence interval [CI], 1.9 to 14.2) and to have been recovered from patients hospitalized in central Canada (OR, 3.0; 95% CI, 1.2 to 7.4). There has been no evidence of vancomycin "MIC creep" in Canadian strains of methicillin (meticillin)-resistant S. aureus, and hVISA strains are currently uncommon.


Automatic detection and quantification of growth spurts.
Growth phenomena are often nonlinear and may contain spurts, characterized by a local increase in the rate of growth. Because measurement error and noise may produce apparent spurts, it is important to identify systematic and reliable spurts. We describe a system, automatic maxima detection (AMD), for statistically identifying significant spurts and computing (1) point of maximal velocity, when the spurt was most intense; (2) start, when the spurt started; (3) amplitude, the intensity of the spurt; and (4) duration, the length of the spurt. We also introduce a software implementation of AMD in MATLAB. In growth of height data, AMD showed a reliable pubertal growth spurt for most children and a reliable prepubertal spurt for some children. In simulated growth of vocabulary, AMD showed a large global spurt and several minispurts. In real vocabulary growth, AMD showed a few spurts. Advantages of AMD include improvements in objectivity, automaticity, quantification, and comprehensiveness.


The evolution of catalytic function in the HIV-1 protease.
The evolution of species is a complex phenomenon based on the optimization of a multidimensional function referred to as fitness. At the level of biomolecular evolution, the fitness function can be reduced to include physiochemical properties relevant to the biological function of a particular molecule. In this work, questions involving the physical-chemical mechanisms underlying the evolution of HIV-1 protease are addressed through molecular simulation and subsequent analysis of thermodynamic properties related to the activity of the enzyme. Specifically, the impact of 40 single amino acid mutations on the binding affinity toward the matrix/capsid (MA/CA) substrate and corresponding transition state intermediate has been characterized using a molecular mechanics Poisson-Boltzmann surface area approach. We demonstrate that this approach is capable of extracting statistically significant information relevant to experimentally determined catalytic activity. Further, no correlation was observed between the effect of mutations on substrate and transition state binding, suggesting independent evolutionary pathways toward optimizing substrate specificity and catalytic activity. In addition, a detailed analysis of calculated binding affinity data suggests that ground-state destabilization (reduced binding affinity for the substrate) could be a contributing factor in the evolutionary optimization of HIV-1 protease. A numerical model is developed to demonstrate that ground-state destabilization is a valid mechanism for activity optimization given the high concentrations of substrate experienced by the functional enzyme in vivo.


Both excitatory and inhibitory neurons transiently form clusters at the outermost region of the developing mammalian cerebral neocortex.
During development of the mammalian cerebral neocortex, postmitotic excitatory neurons migrate toward the outermost region of the neocortex. We previously reported that this outermost region is composed of densely packed relatively immature neurons; we named this region, which is observed during the late stage of mouse neocortical development, the "primitive cortical zone (PCZ)." Here, we report that postmigratory immature neurons spend about 1-1.5 days in the PCZ. An electron microscopic analysis showed that the neurons in the PCZ tend to be in direct contact with each other, mostly in a radial direction, forming "primitive neuronal clusters" with a height of 3-7 cells and a width of 1-2 cells. A time-course analysis of fluorescently labeled neurons revealed that the neurons took their positions within the primitive clusters in an inside-out manner. The neurons initially participated in the superficial part of the clusters, gradually shifted their relative positions downward, and then left the clusters at the bottom of this structure. GABAergic inhibitory interneurons were also found within the primitive clusters in the developing mouse neocortex, suggesting that some clusters are composed of both excitatory neurons and inhibitory interneurons. Similar clusters were also observed in the outermost region of embryonic day (E) 78 cynomolgus monkey occipital cortex and 23 gestational week (GW) human neocortices. In the primate neocortices, including human, the presumptive primitive clusters seemed to expand in the radial direction more than that observed in mice, which might contribute to the functional integrity of the primate neocortex.


Texture discrimination based upon an assumed stochastic texture model.
A new approach to texture discrimination is described. This approach is based upon an assumed stochastic model for texture in imagery and is an approximation to the statistically optimum maximum likelihood classifier. The construction and properties of the stochastic texture model are described and a digital filtering implementation of the resulting maximum likelihood texture discriminant is provided. The efficacy of this approach is demonstrated through experimental results obtained with simulated texture data. A comparison is provided with more conventional texture discriminants under identical conditions. The implications to texture discrimination in realworld imagery are discussed.


Modulatory effect of Ganoderma lucidum on expression of xenobiotic enzymes, oxidant-antioxidant and hormonal status in 7,12-dimethylbenz(a)anthracene-induced mammary carcinoma in rats.
Mushrooms are an important natural source represents a major and untapped potent pharmaceutical product. Ganoderma lucidum (G. lucidum) an important medicinal mushroom has been shown to contain high amount of antioxidant. However, in vivo studies on G. lucidum fruiting bodies are lacking. To determine the effects of G. lucidum fruiting bodies ethanolic extract (GLEet) on expression of xenobiotic enzymes, oxidant-antioxidant and hormonal status on 7,12-dimethyl benz[a]antheracene (DMBA) induced experimental breast cancer was investigated in female Sprague dawley rats. Cancer bearing female Sprague dawley rats was orally treated with GLEet (500mg/kg body weight) for 16 weeks. Incidence and tumor volume in each groups, and biochemical parameters were carried out in plasma, liver, and mammary tissues of animals. Histopathological and immunohistochemical analysis were also determined. Oral administration of GLEet on tumor bearing animals significantly diminished the levels of lipid peroxidation thereby enhancing the nonenzymatic antioxidants and also positively regulated the estrogen receptor hormones level to near normal when compared with DMBA treated rats. Moreover, it also positively modulates the xenobiotic metabolizing enzymes. Therefore, the dietary administration of G. lucidum may be efficiently used as a chemopreventive agent against mammary carcinogenesis. We concluded that G. lucidum is a potent chemopreventive agent, thereby it offers maximum protection against DMBA-induced mammary carcinogenesis.


The depolarization - attenuated backscatter relation: CALIPSO lidar measurements vs. theory.
Using measurements obtained by the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observations (CALIPSO) satellite, relationships between layer-integrated depolarization ratio (delta) and layer-integrated attenuated backscatter (gamma) are established for moderately thick clouds of both ice and water. A new and simple form of the delta-gamma relation for spherical particles, developed from Monte Carlo simulations and suitable for both water clouds and spherical aerosol particles, is found to agree well with the observations. A high-backscatter, low-depolarization delta-gamma relationship observed for some ice clouds is shown to result primarily from horizontally oriented plates and implies a preferential lidar ratio - depolarization ratio relation in nature for ice cloud particles containing plates.


A candidate molecule approach to defining developmental pathology in schizophrenia.
The evidence that schizophrenia may have its origins from early in life, possibly during prenatal brain development, is based primarily on a constellation of nonspecific anatomical findings and on the results of surveys of obstetrical complications and of childhood neurological and psychological adjustment. The developmental processes implicated by this evidence are uncertain, but speculation has centered around abnormalities of neuronal proliferation, migration, and connection formation. These developmental milestones are the results of complicated cellular processes involving molecular interactions between cells and between the extracellular and intracellular milieus. To understand how these abnormalities could relate to schizophrenia, it is necessary to characterize the molecular events that define the processes. In this article, we discuss the potential impact of a number of molecules that are important in the sequence of cellular events implicated in schizophrenia. In particular, we focus on molecular mechanisms related to cell proliferation, axonal outgrowth, cell migration, cell survival, synaptic regression, myelination, and developmental aspects of early adult life. These various candidate molecules regulate different aspects of cell growth and cell-cell interactions and are involved in the regulation of deoxyribonucleic acid (DNA) expression. Very few of these molecules have been studied in the schizophrenic brain.


Spatio-temporal correlations in human gamma band electrocorticograms.
Animal electrocorticogram (ECoG) studies have shown that spatial patterns in the gamma band (>20 Hz) reflect perceptual categorization. Spatio-temporal correlations were investigated in the 20-50 Hz range in search for similar phenomena in human ECoG. ECoGs were recorded in a somatosensory discrimination task from 64-electrode subdural grid arrays, with inter-electrode spacing of 1 cm, overlying somatosensory, motor and superior temporal cortices in 2 patients with intractable epilepsy. Bootstrap techniques were devised to analyze the spatial and temporal characteristics of the correlations. Despite an extensive search, no evidence was found for globally correlated activity related to behavior either in narrow (1.e., 35-45 Hz) or broad (i.e., 20-50 Hz) bands. Spatial patterns, extracted using principal component analysis, could not be classified with respect to stimulus type in any time interval. Instead, spatially and temporally intermittent synchronization was observed between pairs of electrodes in 1 cm X 1 cm regions with high variability within and across trials. The distribution of correlation coefficients differed substantially from background levels at inter-electrode distances of 1 cm and 1.4 cm but not 2 cm or more. The minimum duration of correlation, the decorrelation time, of the ECoG was about 50 msec; the average correlation duration at 1 cm inter-electrode distance was about 150 msec; and the recurrence rate of significant correlation peaks was about 1.3/sec. The findings suggest that the surface diameters of domains of spatially correlated activity underlying perceptual categorization in human gamma band ECoG are limited to less than 2 cm and that the intermittent synchronization observed across separations of 1 cm and 1.4 cm is not solely due to volume conduction. Thus, if such gamma band spatial patterns exist in the human brain, no existing technology would be capable of measuring them at the scalp, and subdural electrode arrays for cortical surface recording would have to have spacings under 5 mm.


Extraction of green labeled pectins and pectic oligosaccharides from plant byproducts.
Green labeled pectins were extracted by an environmentally friendly way using proteases and cellulases being able to act on proteins and cellulose present in cell walls. Pectins were isolated from different plant byproducts, i.e., chicory roots, citrus peel, cauliflower florets and leaves, endive, and sugar beet pulps. Enzymatic extraction was performed at 50 degrees C for 4 h, in order to fulfill the conditions required for microbiological safety of extracted products. High methoxy (HM) pectins of high molar mass were extracted with three different enzyme mixtures. These pectins were subsequently demethylated with two pectin methyl esterases (PMEs), either the fungal PME from Aspergillus aculeatus or the orange PME. It was further demonstrated that high molar mass low methoxy (LM) pectins could also be extracted directly from cell walls by adding the fungal PME to the mixture of protease and cellulase. Moreover, health benefit pectic oligosaccharides, the so-called modified hairy regions, were obtained after enzymatic treatment of the residue recovered after pectin extraction. The enzymatic method demonstrates that it is possible to convert vegetable byproducts into high-added value compounds, such as pectins and pectic oligosaccharides, and thus considerably reduce the amount of these residues generated by food industries.


HIV Rev self-assembly is linked to a molten-globule to compact structural transition.
By regulating the differential expression of proviral pre mRNA in the host cell, Rev plays a crucial role in the HIV-1 life cycle. The capacity of Rev to function is intimately linked to its ability to self-associate. Nevertheless, little is known about the exact role of self-association in the molecular mechanism defining its biological activity. A prerequisite knowledge is a definition of the molecular events undertaken by Rev during the process of self-assembly. Thus, this study was initiated to monitor the structure of Rev as a function of protein concentration. Rev undergoes a structural transition as a consequence of self-assembly. This structural transition was monitored by three spectroscopic methods. The accessibility of the single tryptophan in Rev monomer to acrylamide quenching increases with decreasing protein concentration. At very low concentration of Rev, the tryptophan accessibility is close to that of an unfolded Rev. As evaluated by circular dichroism, the secondary structure of Rev is protein concentration dependent as evidenced by an increase in the magnitude of ellipticity with increasing protein concentration. Further, results from ANS binding studies indicate that the ANS binding sites in Rev experience an apparent increase in hydrophobicity as the Rev concentration was increased. These concentration dependent changes seem to reach a maximum above 5 microM Rev monomer concentration. In order to define the mode of Rev self-association sedimentation velocity and equilibrium experiments were conducted. There are evidently two consecutive progressive association processes. At protein concentrations below 0.5 mg/ml, the data from sedimentation studies can be fitted to a single isodesmic model. Simulation of velocity sedimentation profile indicates that free Rev monomer that has not entered into the association processes can best be described to exhibit a value of S(20,w) that is substantially smaller than 1.4 S, a value needed to fit the rest of the data. The latter value is consistent for a Rev monomer with the expected molecules weight and if it were to assume a compact globular shape. These spectroscopic and hydrodynamic results imply that monomeric Rev is in a molten globule state, which becomes more compact upon self-association.


[Thiamine diphosphate level and metabolism in allergic myocarditis and treatment with peloid].
Enzymatic systems involved in thiamin metabolism were studied in experimental allergic myocarditis. Development of inflammation in myocardium was accompanied by a distinct activation of thiamin pyrophosphatase which catalyzed the most important step responsible for deterioration of the coenzyme functions. The stabilizing effect of peloid on the intracellular pool and compartmentalization of thiamin in myocarditis involved equilibration of the anabolic and catabolic reactions in the coenzyme metabolism, mainly due to selective inhibition of the thiamin pyrophosphatase activity stimulated under these conditions.


Aortic pseudoaneurysm due to ingested foreign body.
We have described a child with pseudoaneurysm of the thoracic aorta caused by an ingested safety pin, who survived this life-threatening injury in which the diagnosis was elusive for four months. Esophageal foreign bodies, especially sharp, pointed ones, can cause such a cardiovascular complication. A high index of suspicion is imperative if the diagnosis is to be made early. Serial chest x-ray films showing an enlarging mediastinal mass should trigger aggressive radiologic evaluation.


Determination and use of haplotypes: ethnic comparison and association of the lipoprotein lipase gene and coronary artery disease in Mexican-Americans.
To illustrate an approach of deriving haplotypes for genetic association studies, using the lipoprotein lipase (LPL) gene and coronary artery disease. Six polymorphisms sufficient to distinguish the most common haplotypes in the 3' end of LPL were identified by genotyping 10 polymorphisms in a small pilot population. These were used to haplotype LPL in large family samples of Mexican-Americans and non-Hispanic Caucasians. A case-control association study was performed comparing Mexican-Americans with and without coronary artery disease. The two ethnic groups exhibited significant genetic differences. Among Mexican-Americans, homozygosity for LPL haplotype 1 was protective against coronary artery disease (OR = 0.50, 95% CI 0.27-0.91). This study outlines the haplotype structure of the LPL gene, illustrates the utility of haplotype-based analysis in association studies, and demonstrates the importance of defining haplotype frequencies for different ethnic groups.


Systemic hypertension and nondiabetic chronic kidney disease: the best evidence-based therapeutic approach today.
Despite the high prevalence and significant morbidity and mortality associated with high chronic kidney disease (CKD) in patients with hypertension, it remains vastly under-diagnosed and under-treated. Consequently, many patients develop kidney failure requiring dialysis or kidney transplant. Moreover, patients with CKD represent the group at highest risk from cardiovascular complications, even greater than patients with diabetes mellitus. Therefore, management of hypertension in such patients needs to be more aggressive compared to those with normal kidney function. This review provides guidelines for treatment of hypertension in patients with non-diabetic CKD based on updated evidence from clinical trials data. Following these recommendations is likely to minimize the risk of development of kidney failure and cardiovascular disease.


taxalogue: a toolkit to create comprehensive CO1 reference databases.
Taxonomic identification through DNA barcodes gained considerable traction through the invention of next-generation sequencing and DNA metabarcoding. Metabarcoding allows for the simultaneous identification of thousands of organisms from bulk samples with high taxonomic resolution. However, reliable identifications can only be achieved with comprehensive and curated reference databases. Therefore, custom reference databases are often created to meet the needs of specific research questions. Due to taxonomic inconsistencies, formatting issues, and technical difficulties, building a custom reference database requires tremendous effort. Here, we present taxalogue, an easy-to-use software for creating comprehensive and customized reference databases that provide clean and taxonomically harmonized records. In combination with extensive geographical filtering options, taxalogue opens up new possibilities for generating and testing evolutionary hypotheses. taxalogue collects DNA sequences from several online sources and combines them into a reference database. Taxonomic incongruencies between the different data sources can be harmonized according to available taxonomies. Dereplication and various filtering options are available regarding sequence quality or metadata information. taxalogue is implemented in the open-source Ruby programming language, and the source code is available at https://github.com/nwnoll/taxalogue. We benchmark four reference databases by sequence identity against eight queries from different localities and trapping devices. Subsamples from each reference database were used to compare how well another one is covered. taxalogue produces reference databases with the best coverage at high identities for most tested queries, enabling more accurate, reliable predictions with higher certainty than the other benchmarked reference databases. Additionally, the performance of taxalogue is more consistent while providing good coverage for a variety of habitats, regions, and sampling methods. taxalogue simplifies the creation of reference databases and makes the process reproducible and transparent. Multiple available output formats for commonly used downstream applications facilitate the easy adoption of taxalogue in many different software pipelines. The resulting reference databases improve the taxonomic classification accuracy through high coverage of the query sequences at high identities.


Photoperiodic condition modulates the effects of testosterone on song control nuclei volumes in male European starlings.
Songbirds possess a specialized network of brain nuclei mediating song learning and production, the song system. In many species, the nuclei in this circuit are larger in volume in males housed on long photoperiods than in males housed on short photoperiods. Exposure to long days initially leads to increases in circulating levels of testosterone (T) and it is generally believed that volume changes in the song system are controlled by T-dependent effects of photoperiod. However, a growing body of evidence indicates that photoperiod can also have T-independent effects on song system morphology. Previous work in our laboratory suggested that photoperiodic condition may modulate the effects of exogenous testosterone on the volumes of song control nuclei in male European starlings (Sturnus vulgaris). In the present report, we tested the hypothesis that photorefractoriness attenuates the effects of T on the volumes of song control nuclei in starlings. One group of long-day photorefractory males (16L:8D) was exposed to exogenous T and two groups of short-day photosensitive birds (8L:16D) were either implanted with T or blank (control) capsules. After 8 weeks, T levels were elevated in both groups of T-implanted birds and were undetectable in control animals. Volumes of the high vocal center were larger in T-implanted photosensitive birds than in both T-implanted photorefractory and control photosensitive males, which did not differ. These results suggest that photorefractoriness renders the song nuclei of starlings less sensitive to stimulatory effects of T. Plasticity in the starling song system appears to require the coordination of the appropriate hormonal milieu with a permissive photoperiodic condition.


Effect of familial diabetes status and age at diagnosis on type 2 diabetes risk: a nation-wide register-based study from Denmark.
We assessed whether the risk of developing type 2 diabetes and the age of onset varied with the age at diabetes diagnosis of affected family members. We performed a national register-based open cohort study of individuals living in Denmark between 1995 and 2012. The population under study consisted of all individuals aged 30 years or older without diagnosed diabetes at the start date of the cohort (1 January 1995) and who had information about their parents' identity. Individuals who turned 30 years of age during the observation period and had available parental identity information were also added to the cohort from that date (open cohort design). These criteria restricted the study population mostly to people born between 1960 and 1982. Multivariable Poisson regression models adjusted for current age and highest educational attainment were used to estimate incidence rate ratios (IRRs) of type 2 diabetes. We followed 2,000,552 individuals for a median of 14 years (24,034,059 person-years) and observed 76,633 new cases of type 2 diabetes. Compared with individuals of the same age and sex who did not have a parent or full sibling with diabetes, the highest risk of developing type 2 diabetes was observed in individuals with family members diagnosed at an early age. The IRR was progressively lower with a higher age at diabetes diagnosis in family members: 3.9 vs 1.4 for those with a parental age at diagnosis of 50 or 80 years, respectively; and 3.3 vs 2.0 for those with a full sibling's age at diagnosis of 30 or 60 years, respectively. People with a family member diagnosed with diabetes at an earlier age are more likely to develop diabetes and also to develop it at an earlier age than those with a family member diagnosed in later life. This finding highlights the importance of expanding our understanding of the interplay between genetic diabetes determinants and the social, behavioural and environmental diabetes determinants that track in families across generations. Accurate registration of age at diagnosis should form an integral part of recording a diabetes family history, as it provides easily obtainable and highly relevant detail that may improve identification of individuals at increased risk of younger onset of type 2 diabetes. In particular, these individuals may benefit from closer risk factor assessment and follow-up, as well as prevention strategies that may involve the family.


Hybrid approach combining deep learning and a rule based expert system for concept extraction from prescriptions.
Concept extraction from prescriptions is a very important task that provides a foundation for many of the downstream healthcare applications in decision making across the areas of pharmacovigilance, medication adherence, inventory management, and other matters of value-based care. Although short, these directions can sometimes be complex. With the increase in complexity of direction, it becomes harder to extract various concepts by only rule based expert system. It identifies major concepts like frequency, dosage, duration, etc. from the natural text direction using a combination of rules and deep learning (DL) based methods on a large real world data of a pharmacy chain. The DL module includes a fine-tuned BERT transformer and Gram CNN (Convolutional Neural Network) based NER (Named Entity Recognition) architecture. The proposed method utilizes the domain heuristics along with intelligent labelling and bootstrapping to help DL models extract concepts with high evaluation scores and thus provides a way for carrying out concept extraction using targeted methods instead of one single method. To the best of our knowledge, this is the best performance reported in the literature for concept extraction from doctor's prescription.


Maternal Circulating Vitamin Status and Colostrum Vitamin Composition in Healthy Lactating Women-A Systematic Approach.
Colostrum is the first ingested sole nutritional source for the newborn infant. The vitamin profile of colostrum depends on the maternal vitamin status, which in turn is influenced by diet and lifestyle. Yet, the relationship between maternal vitamin status and colostrum vitamin composition has not been systematically reviewed. This review was conducted with the aim to generate a comprehensive overview on the relationship between maternal serum (plasma) vitamin concentration and corresponding colostrum composition. Three electronic databases, Embase (Ovid), Medline (Ovid), and Cochrane, were systematically searched based on predefined inclusion and exclusion criteria. Finally, a total of 11 eligible publications were included that examined the vitamins A, C, D, E, and K in both biological fluids. Maternal vitamin A, D, E, and K blood levels were unrelated to colostrum content of the respective vitamins, and serum vitamin A was inversely correlated with colostrum vitamin E. Colostrum versus maternal serum vitamins were higher for vitamins A, C, and K, lower for vitamin D, and divergent results were reported for vitamin E levels. Colostrum appears typically enriched in vitamin A, C, and K compared to maternal serum, possibly indicative of active mammary gland transport mechanisms. Inter-individual and inter-study high variability in colostrum's vitamin content endorses its sensitivity to external factors.


[Smoking prevention policies in European companies].
Many companies are exploring the issue of adopting measures to deal with involuntary smoking. The most comprehensive policies aim at systematically taking action in all possible directions (informing employees, stipulating specific smoking areas, treatment for quitting smoking...). Many agencies are likewise carrying out initiatives for intervention at companies. This study reviews interventions currently being carried out and their components, based on company studies from different European countries. The main common principles for a smoking prevention policy with the greatest prospects for success are identified. This information can be used as a reference for a European model for policies on smoking at the workplace.


Characterization of structural properties for morphological differentiation of melanosomes: II. Electron microscopic and SDS-PAGE comparison of melanosomal matrix proteins in B16 and Harding Passey melanomas.
This study compared the qualitative and quantitative differences of the melanosomal matrix proteins between the ellipsoidal-lamellar melanosomes of B16 melanoma and the spherical-granular melanosomes of Harding Passey (HP) melanoma by electron microscopy and SDS (sodium dodecyl sulfate)-PAGE (polyacrylamide gel electrophoresis). The purity of the melanosomes at each step of the sample preparations was assayed by % distribution of cytoplasmic marker enzymes and proteins. Characterization of the melanosomal matrix was carried out in material purified by the 2 steps of sucrose density gradient ultracentrifugation and membrane dissociation with BRIJ-35. Eighty-three percent of the matrix proteins were solubilized from both B16 and HP by guanidine HCl (GH), which also caused marked ultrastructural changes in different ways. The inner matrix of HP completely disintegrated into fine grains whereas the matrix of B16 showed a great loss in electron density and was degraded, though its basic framework remained unchanged. Under SDS-PAGE, the melanosomal matrix was dissociated into 14 polypeptide bands with respect to size and charge density, 10 of which (63-85% of total proteins solubilized by GH) were common to both B16 and HP. A significant quantitative difference was noted in the relative amounts of these common proteins. The remaining 4 were unique in molecular weight to each form of melanosome though their relative amounts were low. It is suggested that these chemical differences, i.e., quantitative and qualitative, of the matrix proteins are responsible for the ultrastructural differentiation of melanosomes.


Mobilization and redistribution of default mode network from resting state to task state in amnestic mild cognitive impairment.
Brain task-negative networks (default mode network, DMN) and task-positive networks appear to operate largely in opposition, such that task-negative networks show activation during resting states, whilst task-positive networks are deactivated with the reverse being true during goal-oriented behavior. Altered DMN and task-positive network activity has been observed in amnestic mild cognitive impairment (aMCI) subjects. However, no study has directly linked the patterns of between-state differences in the same aMCI cohort regarding these two types of functional networks. The spatial and temporal characteristics of intrinsic, low frequency BOLD signal fluctuations both during resting state and episodic memory fMRI task were assessed in 28 aMCI subjects and 23 matched healthy controls, using a posterior cingulate cortex- based temporal correlation analysis. aMCI subjects showed impaired attenuation in the DMN between rest and task state, and greater cognitive impairment was associated with decreased ability to attenuate DMN during task engagement. Moreover, more redistributed resource from DMN appeared to be required in aMCI to maintain the similar task performance possibly to offset their inability to engage task-positive networks. In order to complete a given task, mobilized and redistributed resources of DMN appeared to replace task-positive network function to some degrees in aMCI subjects. This may represent an inability to control the switching of functional modes between these types of network.


Transcription rates in DNA brushes.
We theoretically predict the rate of transcription (TX) in DNA brushes by introducing the concept of TX dipoles that takes into account the unidirectional motion of enzymes (RNAP) along DNA during transcription as correlated pairs of sources and sinks in the relevant diffusion equation. Our theory predicts that the TX rates dramatically change upon the inversion of the orientation of the TX dipoles relative to the substrate because TX dipoles modulate the concentrations of RNAP in the solution. Comparing our theory with experiments suggests that, in some cases, DNA chain segments are relatively uniformly distributed in the brush, in contrast to the parabolic profile expected for flexible polymer brushes.


[Medical communication about the management of depression, anxiety and sleeplessness in the Dutch women's magazine 'Margriet' between 1950 and 1960].
The reporting of health and medicine in the media played an important role in the way people perceived, defined and coped with everyday health problems in the second half of the twentieth century. It was and still is predominantly a supply-market which is dominated by the medical approach; creating a medical aura of progress and the self-evidence of a cure. Although the medical approach figured also prominently in women's magazines, the reporting of illness and health is far more a product of supply and demand with a lively interaction between readers and the editorial office by the means of topic-related letter columns. As such women's magazines not only offer a gender-specific but also a more balanced source for acquiring a better understanding of how public definitions and perceptions of illness and health changed over time. In this article we will focus on the communication about the management of health problems related to depression, anxiety and sleeplessness in the prototypical Dutch women's magazine 'Margriet' between 1950 and 1960. Our guiding research question has been: how do notions about depression, anxiety and and about responsive health behaviour, of which psychotropic drug use is a part, change over time in the reporting of health problems in Margriet? This question is of particular interest to learn more about the historical dynamics of the culture- and gender-specific public interplay between patients and doctors in terms of conceptualising the aforementioned health problems and defining medical coping strategies. Among other things we show that although there was hardly any mention of any 'functional division of labour' between mind and body in 1950, the mind-body dichotomy started to play an important role in the way health problems were perceived in 1960.


Compensatory mutations, antibiotic resistance and the population genetics of adaptive evolution in bacteria.
In the absence of the selecting drugs, chromosomal mutations for resistance to antibiotics and other chemotheraputic agents commonly engender a cost in the fitness of microorganisms. Recent in vivo and in vitro experimental studies of the adaptation to these "costs of resistance" in Escherichia coli, HIV, and Salmonella typhimurium found that evolution in the absence of these drugs commonly results in the ascent of mutations that ameliorate these costs, rather than higher-fitness, drug-sensitive revertants. To ascertain the conditions under which this compensatory evolution, rather than reversion, will occur, we did computer simulations, in vitro experiments, and DNA sequencing studies with low-fitness rpsL (streptomycin-resistant) mutants of E. coli with and without mutations that compensate for the fitness costs of these ribosomal protein mutations. The results of our investigation support the hypothesis that in these experiments, the ascent of intermediate-fitness compensatory mutants, rather than high-fitness revertants, can be attributed to higher rates of compensatory mutations relative to that of reversion and to the numerical bottlenecks associated with serial passage. We argue that these bottlenecks are intrinsic to the population dynamics of parasitic and commensal microbes and discuss the implications of these results to the problem of drug resistance and adaptive evolution in parasitic and commmensal microorganisms in general.


Observing Phase Transition of a Temperature-Responsive Polymer Using Electrochemical Collisions on an Ultramicroelectrode.
Herein, a study on a new lower critical solution temperature (LCST) polymer in an organic solvent by an electrochemical technique has been reported. The phase-transition behavior of poly(arylene ether sulfone) (PAES) was examined on 1,2-dimethoxyethane (DME). At a temperature above the LCST point, polymer molecules aggregated to create polymer droplets. These droplets subsequently collided with an ultramicroelectrode (UME), resulting in a new form of staircase current decrease. The experimental collision frequency and collision signal were analyzed in relation to the concentration of the polymer. In addition, the degree of polymer aggregation associated with temperature change was also observed.


Local breast cancer recurrence caused by mammographically guided punctures.
To evaluate the risk of needle track seeding or tumor cell implantation as the cause of locally recurrent breast cancer after breast conserving surgery. We reviewed recurrences from a consecutive series of 303 clinically nonpalpable breast cancers treated with breast conserving surgery after pre-operative localization. The median mammographic follow-up was 5.4 years. The suspicion of seeding or implantation was based on the location of the recurrent lesion in comparison with the needle path in two orthogonal mammographic projections. Pre-operative percutaneous biopsies had been done in 71% (214/303) of the cases. Postoperative radiotherapy was administered to 82% (194/238) of the invasive cancers and to 28% (18/65) of the ductal cancers in situ (DCIS). Locally recurrent cancer occurred in 11% (33/303) of the cases. Radiotherapy demonstrated a protective effect from relapse among invasive cancers but not for DCIS. Seeding or implantation was suspected in 3 recurrent invasive cancers which had not been subject to radiotherapy. The histopathological diagnosis of the primary cancer and the recurrent cancer were the same in these cases: adenoid cystic, mucinous and tubuloductal cancer. Seeding or implantation was suspected as the cause of local recurrence in 7% (3/44) of the invasive cancers which did not receive radiotherapy.


Spatial self-organized patterning in seagrasses along a depth gradient of an intertidal ecosystem.
The spatial structure of seagrass landscapes is typically ascribed to the direct influence of physical factors such as hydrodynamics, light, and sediment transport. We studied regularly interspaced banded patterns, formed by elongated patches of seagrass, in a small-scale intertidal ecosystem. We investigated (1) whether the observed spatial patterns may arise from feedback interactions between seagrass and its abiotic environment and (2) whether changes in abiotic conditions may lead to predictable changes in these spatial patterns. Field measurements, experiments, and a spatially explicit computer model identified a "scale-dependent feedback" (a mechanism for spatial self-organization) as a possible cause for the banded patterns. Increased protection from uprooting by improved anchoring with increasing seagrass density caused a local positive feedback. Sediment erosion around seagrass shoots increased with distance through the seagrass bands, hence causing a long-range negative feedback. Measurements across the depth gradient of the intertidal, together with model simulations, demonstrated that seagrass cover and mean patch size were predictably influenced by additional external stress caused by light limitation and desiccation. Thus, our study provides direct empirical evidence for a consistent response of spatial self-organized patterns to changing abiotic conditions, suggesting a potential use for self-organized spatial patterns as stress indicators in ecosystems.


Ciliary body thickness and refractive error in children.
To determine whether ciliary body thickness (CBT) is related to refractive error in school-age children. Fifty-three children, 8 to 15 years of age, were recruited. CBT was measured from anterior segment OCT images (Visante; Carl Zeiss Meditec, Inc., Dublin, CA) at 1 (CBT1), 2 (CBT2) and 3 (CBT3) mm posterior to the scleral spur. Cycloplegic refractive error was measured with an autorefractor, and axial length was measured with an optical biometer. Multilevel regression models determined the relationship between CBT measurements and refractive error or axial length. A Bland-Altman analysis was used to assess the between-visit repeatability of the ciliary body measurements. The between-visits coefficients of repeatability for CBT1, -2, and -3 were 148.04, 165.68, and 110.90, respectively. Thicker measurements at CBT2 (r = -0.29, P = 0.03) and CBT3 (r = -0.38, P = 0.005) were associated with increasingly myopic refractive errors (multilevel model: P < 0.001). Thicker measurements at CBT2 (r = 0.40, P = 0.003) and CBT3 (r = 0.51, P < 0.001) were associated with longer axial lengths (multilevel model: P < 0.001). Thicker ciliary body measurements were associated with myopia and a longer axial length. Future studies should determine whether this relationship is also present in animal models of myopia and determine the temporal relationship between thickening of the ciliary muscle and the onset of myopia.


Investigation of Schirmer tear test-1 for measurement of tear production in cats in various environmental settings and with different test durations.
To assess reliability of the Schirmer tear test-1 (STT-1) for measurement of tear production in cats in various environments, investigate whether sympathetic stimulation impacts measurements, and determine whether meaningful conclusions regarding lacrimation in cats can be drawn from STT-1 measurements obtained with STT strip placement for < 1 minute. 176 cats examined in a private practice (n = 100), a feral cat clinic (56), or a veterinary teaching hospital (20). The STT-1 was performed in both eyes of each cat. Measurements were recorded at 10- or 30-second intervals for 1 minute. Cats at the teaching hospital were tested once in a quiet examination room (unstimulated conditions) and once in the same room with loud prerecorded noises (stimulated conditions), with a 30-minute interval between tests and evaluation of cats' heart rates before and after STT-1. Data were analyzed with parametric statistical tools and a nonlinear mixed-effect model. 30- and 60-second STT-1 measurements were significantly correlated (r = 0.94). The STT-1 measurements did not differ under nonstimulated versus stimulated conditions, despite significant changes in heart rates that indicated sympathetic stimulation. A hyperbolic model of STT-1 kinetics was validated, allowing for extrapolation of measurements obtained in < 60 seconds and generation of reference values (95% predictive intervals) for various test durations. Median (95% predictive interval) 30- and 60-second STT-1 measurements were 9.1 mm (4.8 to 15.6 mm) and 14.3 mm (8.2 to 22.3 mm), respectively. The STT-1 was a reliable diagnostic test in all settings; results were not affected by sympathetic stimulation, and a shorter duration of testing could be considered in selected cases.


Prioritizing prevention opportunities in the Washington State construction industry, 2003-2007.
This study compares construction industry groups in Washington State by injury severity and cost, and ranks industry groups according to potential for prevention. All Washington State workers' compensation compensable claims with date of injury between 2003 and 2007 were classified into North American Industrial Classification System (NAICS) industry groups. Claims were then aggregated by injury type and industry groups were ranked according to a prevention index (PI). The PI is the average of the rank orders of the claim count and the claim incidence rate. A lower PI indicates a higher need for prevention activities. The severity rate was calculated as the number of days of time loss per 10,000 full-time equivalents (FTEs). For all injury types, construction industry groups occupy 7 of the top 15 PI ranks in Washington State. The severity rate among construction industry groups was twice that for non-construction groups for all injury types. Foundation, structure, and building exterior contractors (NAICS 2381) ranked highest in prevention potential and severity among construction industry groups for most common injury types including falls from elevation, fall on same level, struck by/against, and musculo-skeletal disorders of the neck, back, and upper extremity (WMSDs). Median claim costs by injury type were generally higher among construction industry groups. The construction industry in Washington State has a high severity rate and potential for prevention. The methods used for characterizing these industry groups can be adapted for comparison within and between other industries and states. These data can be used by industry groups and employers to identify higher cost and higher severity injury types. Knowledge about the relative frequencies and costs associated with different injury types will help employers and construction industry associations make better informed decisions about where prevention efforts are most needed and may have the greatest impact. The results of this study can also be used by industry stakeholders to cooperatively focus on high cost and high severity injuries and explore best practices, interventions, and solutions as demonstrated by efforts to prevent musculoskeletal disorders in masonry (Entzel, Albers, & Welch, 2007). Initiating construction industry groups to focus on high cost and high severity injuries may also help prevent other types of injuries.


Preparation of isolated bovine adipocytes: validation of use for studies characterizing insulin sensitivity and binding.
The present study was undertaken to develop techniques to isolate bovine adipocytes, to compare their rates of glucose metabolism and insulin sensitivity with adipocytes in adipose tissue, and to determine if isolated bovine adipocytes specifically bind insulin. Cell size and diameter distributions were the same for adipocytes fixed with OsO4 after isolation with collagenase and adipocytes liberated from OsO4-fixed adipose tissue slices. On a per cell basis, lipogenic rates were greater for isolated adipocytes compared with intact adipose tissue. Similar differences were found for glucose oxidation. In short term incubations, glucose oxidation and lipid synthesis were not stimulated by insulin (0-100 ng/ml) in either isolated adipocytes or tissue. Specific binding of [125I]iodoinsulin at 30 C was low (0.8%) in the first group of six beef cattle sampled, but increased with increasing cell concentration. Insulin degradation after 90 min was less than 5%. The specificity of [125I]iodoinsulin binding was studied in a second group of six animals. There was no specific binding of insulin in this group. In summary, bovine adipocytes can be isolated which are metabolically active and provide a valid system for studying hormone binding and action. In the present study, glucose metabolism in bovine adipocytes was not stimulated by insulin in vitro. This insensitivity to insulin was associated with a negligible capacity for insulin binding. These findings suggest that the lack of insulin sensitivity in bovine adipose tissue may be due to an inability to specifically bind insulin. This may be related to the unique metabolism of ruminant adipose tissue, which is less dependent upon glucose for fatty acid synthesis than is adipose tissue from nonruminant species.


Lower socioeconomic status and disability among US adults with chronic kidney disease, 1999-2008.
Socioeconomic disparities are associated with the prevalence of disability in the general population; however, it is unknown whether a similar association exists between socioeconomic status and disability from chronic kidney disease (CKD, defined as albuminuria or an estimated glomerular filtration rate of 15-59 mL/min/1.73 m(2)). A total of 4,257 US adults aged 20 years or older with CKD who participated in the National Health and Nutrition Examination Survey 1999-2008 completed standardized questionnaires assessing self-reported difficulties in activities of daily living (ADL), instrumental ADL (IADL), lower-extremity mobility (LEM), and leisure and social activities (LSA). We used multivariable logistic regression with population-based weighting to obtain adjusted prevalence estimates of disability by demographic, socioeconomic, health care access, and clinical characteristics. Participants with less education had more disability (age- and sex-adjusted prevalence of disability by lowest vs highest level of education: ADL, 24.5% vs 16.9%; IADL, 34.0% vs 20.3%; LEM, 56.9% vs 44.6%; LSA, 26.2% vs 16.8%; P < .001 for all). We observed similar trends for income. After further adjustment for other sociodemographic factors, health care access, and comorbid conditions, education and income both remained significantly associated, by any measure, with lower prevalence of disability. Among people with CKD in the United States, lower socioeconomic status is associated with greater risk of disability, independent of race/ethnicity, health care access, and comorbid conditions. Our findings suggest that people with CKD and limited education or lower income should be targeted for early intervention to limit disability and further loss of income, both of which could worsen outcomes in CKD.


Quantification of Cre-mediated recombination by a novel strategy reveals a stable extra-chromosomal deletion-circle in mice.
Inducible conditional knockout animals are widely used to get insight in the function of genes and the pathogenesis of human diseases. These models frequently rely on Cre-mediated recombination of sequences flanked by Lox-P sites. To understand the consequences of gene disruption, it is essential to know the efficiency of the recombination process. Here, we describe a modification of the multiplex ligation-dependent probe amplification (MLPA), called extension-MLPA (eMLPA), which enables quantification of relatively small differences in DNA that are a consequence of Cre-mediated recombination. eMLPA, here applied on an inducible Pkd1 conditional deletion mouse model, simultaneously measures both the reduction of the floxed allele and the increase of the deletion allele in a single reaction thereby minimizing any type of experimental variation. Interestingly, with this method we were also able to observe the presence of the excised DNA fragment. This extra-chromosomal deletion-circle was detectable up to 5 months after activation of Cre. eMLPA is a novel strategy which easily can be applied to measure the Cre-mediated recombination efficiency in each experimental case with high accuracy. In addition the fate of the deletion-circle can be followed simultaneously.


Cost-Effectiveness of Carbidopa-Levodopa Enteral Suspension for Advanced Parkinson's Disease in the United States.
Carbidopa/levodopa enteral suspension (CLES) is indicated for the treatment of advanced Parkinson's disease (aPD) with severe motor fluctuations. To determine the cost, quality-adjusted life years (QALY), and cost-effectiveness of CLES compared to the standard-of-care (SoC) for aPD patients in the United States (US), using real-world data. A published Markov model, comprising of 25 health states and a death state, (defined by a combination of the Hoehn and Yahr scale and waking time spent in OFF-time) was adapted to estimate the benefits for CLES versus oral SoC over a patient's lifetime in the US healthcare setting. Clinical inputs were based on a clinical trial and a registry study; utility inputs were sourced from the Adelphi-Disease Specific Programmes. CLES compared to SoC was associated with incremental costs ($1,031,791 vs. $1,025,180) and QALY gain (4.61 vs. 3.76), resulting in an incremental cost-effectiveness ratio of $7711/QALY. CLES is a cost-effective treatment for aPD patients with medication resistant motor fluctuations. © 2023 AbbVie, Inc and The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.


Endoscopic reverse third ventriculostomy via the cisterna magna: anatomical study and proposal of a novel procedure.
Basilar artery injury has been known as a potential lethal complication of endoscopic third ventriculostomy. In order to avoid this complication, endoscopic reverse third ventriculostomy via a trans-cisterna-magna route was studied. A cadaveric study was performed for navigation of a flexible endoscope through the cisterna magna. Three fresh, unfixed cadavers were used for this endoscopic navigation. In the prone position, a small vertical paramedian skin incision is made at the mid-portion of the posterior neck. An 11-mm threaded plastic tube is inserted towards the posterior arch of the atlas. After a partial hemilaminectomy of the atlas, a flexible endoscope is introduced into the cisterna magna and is navigated cephalad along the vertebrobasilar artery to the inferior aspect of the floor of the third ventricle. Through the working channel of a fiberscope, third ventriculostomy is performed in a reverse direction. Additional detailed anatomy was studied in fixed cadaveric head specimens with a rigid rod-lens endoscope for anatomic orientation. A novel technique of a trans-cisterna-magna reverse third ventriculostomy was studied in cadaveric specimens. This technique may avoid basilar artery injury which occurs occasionally during conventional third ventriculostomy.


Primary myelofibrosis: 2013 update on diagnosis, risk-stratification, and management.
Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by stem cell-derived clonal myeloproliferation, abnormal cytokine expression, bone marrow fibrosis, anemia, splenomegaly, extramedullary hematopoiesis (EMH), constitutional symptoms, cachexia, leukemic progression, and shortened survival. Diagnosis is based on bone marrow morphology. The presence of fibrosis, JAK2/MPL mutation, or +9/13q- cytogenetic abnormality is supportive but not essential for diagnosis. Prefibrotic PMF mimics essential thrombocythemia in its presentation and the distinction is prognostically relevant. Differential diagnosis of myelofibrosis should include chronic myeloid leukemia, myelodysplastic syndromes, chronic myelomonocytic leukemia, and acute myeloid leukemia. The Dynamic International Prognostic Scoring System-plus (DIPSS-plus) prognostic model for PMF can be applied at any point during the disease course and uses eight independent predictors of inferior survival: age >65 years, hemoglobin <10 g/dL, leukocytes >25 × 10⁹/L, circulating blasts ≥ 1%, constitutional symptoms, red cell transfusion dependency, platelet count <100 × 10⁹/L, and unfavorable karyotype (i.e., complex karyotype or sole or two abnormalities that include +8, -7/7q-, i(17q), inv(3), -5/5q-, 12p-, or 11q23 rearrangement). The presence of 0, 1, "2 or 3," and ≥ 4 adverse factors defines low, intermediate-1, intermediate-2, and high-risk disease with median survivals of approximately 15.4, 6.5, 2.9, and 1.3 years, respectively. A >80% two-year mortality is predicted by monosomal karyotype, inv(3)/i(17q) abnormalities, or any two of circulating blasts >9%, leukocytes ≥ 40 × 10⁹/L or other unfavorable karyotype. Most recently, mutations involving ASXL1, SRSF2, EZH2, and IDH1/2 or increased plasma IL-2R, IL-8, or serum-free light chain levels have been shown to adversely affect survival. Observation alone is adequate for asymptomatic low/intermediate-1 risk disease. Allogeneic stem cell transplantation (ASCT) is often considered for high risk disease. Conventional or experimental drug therapy is reasonable for symptomatic intermediate-1 or intermediate-2 risk disease; however, ASCT is an acceptable treatment option for such patients in the presence of ASXL1 or other prognostically adverse mutations. Splenectomy and low-dose radiotherapy are used for drug-refractory splenomegaly. Radiotherapy is also used for the treatment of non-hepatosplenic EMH, PMF-associated pulmonary hypertension, and extremity bone pain.