BAY 41-2272 activates host defence against local and disseminated Candida albicans infections.
In our previous study, we have found that 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]-pyrimidin-4-ylamine (BAY 41-2272), a guanylate cyclase agonist, activates human monocytes and the THP-1 cell line to produce the superoxide anion, increasing in vitro microbicidal activity, suggesting that this drug can be used to modulate immune functioning in primary immunodeficiency patients. In the present work, we investigated the potential of the in vivo administration of BAY 41-2272 for the treatment of Candida albicans and Staphylococcus aureus infections introduced via intraperitoneal and subcutaneous inoculation. We found that intraperitoneal treatment with BAY 41-2272 markedly increased macrophage-dependent cell influx to the peritoneum in addition to macrophage functions, such as spreading, zymosan particle phagocytosis and nitric oxide and phorbol myristate acetate-stimulated hydrogen peroxide production. Treatment with BAY 41-2272 was highly effective in reducing the death rate due to intraperitoneal inoculation of C. albicans, but not S. aureus. However, we found that in vitro stimulation of peritoneal macrophages with BAY 41-2272 markedly increased microbicidal activities against both pathogens. Our results show that the prevention of death by the treatment of C. albicans-infected mice with BAY 41-2272 might occur primarily by the modulation of the host immune response through macrophage activation.


Fabrication and characterization of polyimide-based 'smooth' titanium nitride microelectrode arrays for neural stimulation and recording.
As electrodes are required to interact with sub-millimeter neural structures, innovative microfabrication processes are required to enable fabrication of microdevices involved in such stimulation and/or recording. This requires the development of highly integrated and miniaturized systems, comprising die-integration-compatible technology and flexible microelectrodes. To elicit selective stimulation and recordings of sub-neural structures, such microfabrication process flow can beneficiate from the integration of titanium nitride (TiN) microelectrodes onto a polyimide substrate. Finally, assembling onto cuffs is required, as well as electrode characterization. Flexible TiN microelectrode array integration and miniaturization was achieved through microfabrication technology based on microelectromechanical systems (MEMS) and complementary metal-oxide semiconductor processing techniques and materials. They are highly reproducible processes, granting extreme control over the feature size and shape, as well as enabling the integration of on-chip electronics. This design is intended to enhance the integration of future electronic modules, with high gains on device miniaturization. (a) Fabrication of two electrode designs, (1) 2 mm long array with 14 TiN square-shaped microelectrodes (80 × 80 µm2), and (2) an electrode array with 2 mm × 80 µm contacts. The average impedances at 1 kHz were 59 and 5.5 kΩ, respectively, for the smaller and larger contacts. Both designs were patterned on a flexible substrate and directly interconnected with a silicon chip. (b) Integration of flexible microelectrode array onto a cuff electrode designed for acute stimulation of the sub-millimeter nerves. (c) The TiN electrodes exhibited capacitive charge transfer, a water window of -0.6 V to 0.8 V, and a maximum charge injection capacity of 154 ± 16 µC cm-2. We present the concept, fabrication and characterization of composite and flexible cuff electrodes, compatible with post-processing and MEMS packaging technologies, which allow for compact integration with control, readout and RF electronics. The fabricated TiN microelectrodes were electrochemically characterized and exhibited a comparable performance to other state-of-the-art electrodes for neural stimulation and recording. Therefore, the presented TiN-on-polyimide microelectrodes, released from silicon wafers, are a promising solution for neural interfaces targeted at sub-millimeter nerves, which may benefit from future upgrades with die-electronic modules.


Prevalence and Drug Resistance of Salmonella in Dogs and Cats in Xuzhou, China.
Salmonellosis is a zoonotic disease, and Salmonella spp. can sometimes be found in dogs and cats, posing a risk to human health. In this study, the prevalence and antimicrobial susceptibility of faecal Salmonella were investigated in pet dogs and cats in Xuzhou, Jiangsu Province, China. Faecal samples from 243 dogs and 113 cats, at seven pet clinics, were tested between March 2018 and May 2019. Each Salmonella isolate was characterised using serotyping and antimicrobial susceptibility tests. The prevalence of Salmonella was 9.47% in dogs and 1.77% in cats. Among the 25 isolates, eight serotypes of Salmonella enterica subsp. enterica were detected, S. Kentucky (n = 11), S. Indiana (n = 5), and S. Typhimurium (n = 4) predominating. S. Derby, S. Toucra, S. Sandiego, S. Newport, and S. Saintpaul all occurred singly. The 23 Salmonella strains found in dogs were from seven different serovars, while the two strains in cats were from two. The highest resistance rates were found for tetracycline (92%), azithromycin (88%), cefazolin (84%), nalidixic acid (80%), ampicillin (80%), ceftriaxone (80%), and streptomycin (76%). Resistance to three or more antimicrobial agents was detected in 24 (96%) isolates. Most of the S. Kentucky and S. Indiana isolates were multi-drug resistant to more than 11 agents. The carriage rate was far higher in dogs than in cats from Xuzhou. Some isolated strains were highly resistant to antimicrobials used to treat infections in humans and pets, which may raise the risk of humans being infected with multi-drug resistant Salmonella via close contact with pets.


Making sense of nuclear localization: a zinc-finger protein encoded by a cytoplasmically replicating plant RNA virus acts a transcription factor: a novel function for a member of large family of viral proteins.
Recent studies have uncovered numerous nucleus-localized proteins encoded by plant RNA viruses. Whereas for some of these viruses nuclear (or, more specifically, nucleolar) passage of the proteins is needed for the virus movement within the plant or suppression of host defense, the nuclear function of these proteins remains largely unknown. Recently, the situation has been clarified for one group of plant RNA viruses, the Carlaviruses. Being positive-stranded RNA viruses, carlaviruses multiply exclusively in the cytoplasm. Chrysanthemum virus B (CVB, a carlavirus) encodes a zinc-finger protein p12 targeted to the nucleus in a nuclear localization signal-dependent manner. In a recent work, we demonstrated that p12 directly interacts with chromatin and plant promoters, thus, acts as a eukaryotic transcription factor (TF) and activates expression of a host TF involved in regulation of cell size and proliferation to favor virus infection. Therefore our studies identified a novel nuclear stage of in CVB infection involving modulation of host gene expression and plant development. Whereas it is well established that any RNA virus actively replicating in the cell causes changes in the transcriptome, our study expanded this view by showing that some positive-stranded RNA viruses can directly manipulate host transcription by encoding eukaryotic TFs.


Diurnal periodicity of conidia of aquatic hyphomycetes in water and entrapment on latex-coated slides in two South Indian streams.
Aquatic hyphomycete conidial trapping efficiency by the banyan (F. benghalensis L.) latex-coated glass slides was tested diurnally (3 h intervals) in the Western Ghats (Sampaje) and west coast (Konaje) streams in relation to abiotic factors (humidity, air temperature and water temperature). The conidial trapping efficiency of latex-coated slides was compared with plain glass slides and drift conidia in water. Three methods of assessment showed higher species richness, conidial richness and diversity in Sampaje than in Konaje stream. In both streams, species richness, conidial richness and diversity in latex-coated slides were the highest followed by conidia in water and plain slides. Three-way ANOVA revealed significant differences in overall species and conidial richness between the streams, sampling methods and time of sampling (p < 0.001). Multiple comparisons by Holm-Sidak test revealed significant differences in overall species and conidial richness between Sampaje and Konaje (p < 0.001); latex-coated slides and plain slides (p < 0.001); latex-coated slides and water filtration (p < 0.001); plain slides and water filtration (p < 0.001). Total species, total conidia and diversity assessed by the three methods peaked during 12 am-3 am in Sampaje stream, while during 3 am-6 am in Konaje stream. Cooler conditions due to relatively low water temperature favoured higher diversity of aquatic hyphomycetes in Sampaje than in Konaje stream. The three methods employed in the present study were not biased towards scolecoid or stauroid conidia. The top five species in both streams was composed of both types of conidia corroborating earlier annual or biannual studies in Konaje and Sampaje streams. Thus, assessment of population of aquatic hyphomycetes using banyan latex-coated slides will be advantageous over plain slides and drift conidia in streams.


Evaluation of the heterophil/lymphocyte ratio as a measure of stress in chickens.
The number of lymphocytes in chicken blood samples decreased and the number of heterophils increased in response to stressors and to increasing levels of corticosterone in the chicken feed. The ratio of heterophils to lymphocytes was less variable than the number of heterophil or lymphocyte cells, and the range of values for this ratio was greater than the range of values for heterophils and lymphocytes among control and experimental groups. The heterophil/lymphocyte ratio appears to be a more reliable indicator of levels of corticosterone in the feed and to social stress than were the plasma corticosteroid levels.


Face recognition using kernel direct discriminant analysis algorithms.
Techniques that can introduce low-dimensional feature representation with enhanced discriminatory power is of paramount importance in face recognition (FR) systems. It is well known that the distribution of face images, under a perceivable variation in viewpoint, illumination or facial expression, is highly nonlinear and complex. It is, therefore, not surprising that linear techniques, such as those based on principle component analysis (PCA) or linear discriminant analysis (LDA), cannot provide reliable and robust solutions to those FR problems with complex face variations. In this paper, we propose a kernel machine-based discriminant analysis method, which deals with the nonlinearity of the face patterns' distribution. The proposed method also effectively solves the so-called "small sample size" (SSS) problem, which exists in most FR tasks. The new algorithm has been tested, in terms of classification error rate performance, on the multiview UMIST face database. Results indicate that the proposed methodology is able to achieve excellent performance with only a very small set of features being used, and its error rate is approximately 34% and 48% of those of two other commonly used kernel FR approaches, the kernel-PCA (KPCA) and the generalized discriminant analysis (GDA), respectively.


Solitary spinal eosinophilic granuloma in children.
Solitary vertebral eosinophilic granuloma is said to be a rare spinal condition in children. 'Vertebra plana' is considered as the typical radiological feature of the involved vertebra; but it could be observed only in 40% of the patients. We have studied three pediatric patients with solitary vertebral eosinophilic granuloma. Their complaint was pain and/or stiffness of the back. Two of them developed neurological deficits in their lower limbs. The diagnosis in imaging (plain radiographs, computed tomography scan and magnetic resonance imaging) varied from one case to another. Histopathology confirmed the diagnosis of eosinophilic granuloma. These three patients underwent an open biopsy and a resection of the spinal lesion achieved with or without interfixation. The outcome was satisfactory in these three cases after a mean follow-up period of over 2 years (from 23 to 27 months).


Evaluation of the environmental implications of the incorporation of feed-use amino acids in the manufacturing of pig and broiler feeds using Life Cycle Assessment.
The incorporation of feed-use (FU) amino acids (AAs) in diets results in a reduced use of protein-rich ingredients such as soybean meal, recognized to have elevated contributions to environmental impacts. This study investigated whether the incorporation of L-lysine.HCl, L-threonine and FU-methionine reduces the environmental impacts of pig and broiler feeds using Life Cycle Assessment. The following impact categories were considered: climate change, eutrophication, acidification, terrestrial ecotoxicity, cumulative energy demand and land occupation. Several feeds were formulated either to minimize the cost of the formulation (with or without AA utilization), to maximize AA incorporation (i.e. the cost of AA was considered to be similar to that of soybean meal), or to minimize greenhouse gas emissions. For both pig and broiler feeds, calculations were made first using only cereals and soybean meal as main ingredients and then using cereals and several protein-rich ingredients (soybean meal, rapeseed meal and peas). In addition, these calculations were performed using two types of soybean meal (from Brazil, associated with recent deforestation or not). For broiler feeds, two types of maize (from France, irrigated, with mineral fertilization v. not irrigated, with animal manure fertilization) were also tested. Regarding the feeds formulated to minimize cost, incorporation of AA decreased the values for eutrophication, terrestrial ecotoxicity and cumulative energy demand of both pig and broiler feeds, regardless of the base ingredients. Reduction in climate change and acidification due to the incorporation of AA depended on the nature of the feed ingredients, with the effect of AA incorporation being greater when combined with ingredients with high impacts such as soybean meal associated with deforestation. Feeds formulated to maximize AA incorporation generally had a similar composition to those formulated to minimize cost, suggesting that the costs of AA were not the limiting factor in their incorporation. Feeds formulated to minimize greenhouse gas emissions had the lowest values for climate change and cumulative energy demand, but not for other impacts. Further research is needed to elucidate whether the incorporation of additional AA (tryptophan and valine) along with L-lysine, L-threonine and FU-methionine could decrease on the environmental impacts of pig and broiler feeds further.


Crystal structure of an enhancer of rudimentary homolog (ERH) at 2.1 Angstroms resolution.
The enhancer of rudimentary gene, e(r), of Drosophila melanogaster encodes an enhancer of rudimentary (ER) protein with functions implicated in pyrimidine biosynthesis and the cell cycle. The ER homolog (ERH) is highly conserved among vertebrates, invertebrates, and plants. Xenopus laevis ERH was reported to be a transcriptional repressor. Here we report the 2.1 Angstroms crystal structure of murine ERH (Protein Data Bank ID 1WZ7), determined by the multiwavelength anomalous dispersion (MAD) method. The monomeric structure of ERH comprises a single domain consisting of three alpha-helices and four beta-strands, which is a novel fold. In the crystal structure, ERH assumes a dimeric structure, through interactions between the beta-sheet regions. The formation of an ERH dimer is consistent with the results of analytical ultracentrifugation. The residues at the core region and at the dimer interface are highly conserved, suggesting the conservation of the dimer formation as well as the monomer fold. The long flexible loop (44 approximately 53) is also significantly conserved, suggesting that this loop region may be important for the functions of ERH. In addition, the putative phosphorylation sites are located at the start of the beta2-strand (Thr18) and at the start of the alpha1-helix (Ser24), implying that the phosphorylation might cause some structural changes.


Annexin-enriched osteoblast-derived vesicles act as an extracellular site of mineral nucleation within developing stem cell cultures.
The application of extracellular vesicles (EVs) as natural delivery vehicles capable of enhancing tissue regeneration could represent an exciting new phase in medicine. We sought to define the capacity of EVs derived from mineralising osteoblasts (MO-EVs) to induce mineralisation in mesenchymal stem cell (MSC) cultures and delineate the underlying biochemical mechanisms involved. Strikingly, we show that the addition of MO-EVs to MSC cultures significantly (P < 0.05) enhanced the expression of alkaline phosphatase, as well as the rate and volume of mineralisation beyond the current gold-standard, BMP-2. Intriguingly, these effects were only observed in the presence of an exogenous phosphate source. EVs derived from non-mineralising osteoblasts (NMO-EVs) were not found to enhance mineralisation beyond the control. Comparative label-free LC-MS/MS profiling of EVs indicated that enhanced mineralisation could be attributed to the delivery of bridging collagens, primarily associated with osteoblast communication, and other non-collagenous proteins to the developing extracellular matrix. In particular, EV-associated annexin calcium channelling proteins, which form a nucleational core with the phospholipid-rich membrane and support the formation of a pre-apatitic mineral phase, which was identified using infrared spectroscopy. These findings support the role of EVs as early sites of mineral nucleation and demonstrate their value for promoting hard tissue regeneration.


EZH2-CCF-cGAS Axis Promotes Breast Cancer Metastasis.
Cytoplasmic chromatin fragments (CCF) are recognized by the cytoplasmic DNA sensor cyclic GMP-AMP synthase (cGAS), which activates the cGAS-STING (cyclic GMP-AMP synthase-stimulator of interferon genes) pathway and promotes the production of inflammatory factors and breast cancer metastasis. However, the mechanisms by which CCF are formed in tumor cells and CCF activation cGAS promotes breast cancer metastasis remain unclear. Here, we report that the enhancer of zeste homolog 2 (EZH2) can promote the formation of CCF and activate the cGAS-STING pathway to promote breast cancer metastasis. Further research found that the EZH2-mediated CCF formation depended on high mobility group A1 (HMGA1), while the stability of EZH2 required ubiquitin-specific peptidase 7 (USP7), indicating that the EZH2-HMGA1-USP7 complex regulated CCF formation. Moreover, EZH2 can activate cGAS through CCF, requiring USP7 to deubiquitinate cGAS and stabilize cGAS. In vivo experimental results showed that EZH2 could promote breast cancer metastasis through CCF. Our findings highlight a new target for breast cancer metastasis. Targeting the EZH2-CCF-cGAS axis may be a potential therapeutic strategy for inhibiting breast cancer metastasis.


Outbreak of canine distemper in domestic ferrets (Mustela putorius furo).
In 2006 an outbreak of canine distemper affected 14 young domestic ferrets in Barcelona, Spain. Their clinical signs included a reduced appetite, lethargy, dyspnoea, coughing, sneezing, mucopurulent ocular and nasal discharges, facial and perineal dermatitis, diarrhoea, splenomegaly and fever. Late in the course of the disease, general desquamation and pruritus, and hyperkeratotic/crusting dermatitis of the lips, eyes, nose, footpads, and perineal area were observed. None of the ferrets developed neurological signs. Non-regenerative anaemia and high serum concentrations of alpha- and beta-globulins were the most common laboratory findings. Most of the animals died or were euthanased because of respiratory complications. Postmortem there were no signs of lung collapse. Distemper was diagnosed by direct immunofluorescence of conjunctival swabs or pcr of several organs, and histology revealed the characteristic eosinophilic intracytoplasmic and intranuclear inclusion bodies of canine distemper virus in several organs. The minimum incubation periods calculated for six of the ferrets were 11 to 56 days, and in 13 of the ferrets the signs of disease lasted 14 to 34 days. Inclusion bodies compatible with infection by herpesvirus were found in the lungs of one of the ferrets.


P26-based serodiagnosis for Bartonella spp. infection in cats.
Bartonella henselae P26 has been identified as an immunodominant antigen expressed during feline infection. We used antisera from cats experimentally infected with B. henselae (n = 6), B. clarridgeiae (n = 4), or B. koehlerae (n = 2) and from a sample of naturally infected cats (B. henselae, n = 34; B. clarridgeiae, n = 1) to evaluate recombinant P26 (rP26) as a serodiagnostic antigen. Immunoblots using antisera from cats infected with B. henselae and B. clarridgeiae reacted strongly with rP26, whereas B. koehlerae antisera did not. A capture ELISA was designed to evaluate the kinetics of rP26 IgG in sera from experimentally infected cats. For B. henselae and B. clarridgeiae antisera, the kinetic profiles of reactivity were similar for rP26 capture ELISA and Bartonella spp. indirect fluorescence assay. However, for B. koehlerae antisera, reactivity in rP26 capture ELISA was consistently low. The serodiagnostic potential of rP26 capture ELISA was evaluated using sera from cats with known Bartonella sp. exposure histories. All 24 (100%) uninfected cats were seronegative, and 33 of 35 (94.3%) cats bacteremic for Bartonella spp. were seropositive. We propose that rP26-based serology can serve as a useful adjunct tool for the diagnosis of feline infection with B. henselae and B. clarridgeiae, but it may not be useful for feline infection with B. koehlerae.


Xanthium strumarium seed extract boosts osteogenesis in human dental pulp stem cell model.
In traditional medicine, Xanthium strumarium is used as an anti-inflammatory and anti-arthritic plant-based medicine. Human Dental Pulp Stem Cells (hDPSCs) are an ideal in vitro model for drug and bioactive compound screening. This study assessed the potential of X. strumarium aqueous extract on hDPSCs differentiation towards the osteogenic lineage. HDPSCs were isolated and cultured by explant method and characterized by surface marker expression, Colony Forming units fibroblasts (CFU-F), Population Doubling time (PDT), and tri-lineage differentiation. X. strumarium aqueous seed extract (XSE) was prepared and its cytotoxic effect on hDPSCs was examined by MTT assay. The effect of XSE on hDPSC differentiation into osteocytes was investigated by biochemical staining and gene expression. The hDPSCs were positive for CD73, CD90, and CD105 and negative for CD34, CD45, and HLA-DR surface markers. The cells had a colony-forming ability with a PDT of 44.91 h. The hDPSCs differentiated into osteocytes, chondrocytes, and adipocytes. The XSE concentration of 15 μg/ml had a significant increase in hDPSC viability. Alizarin Red S staining revealed that XSE treatment enhanced calcium accumulation and matrix mineralization in hDPSCs. XSE treatment also increased osteonectin and IL-6 transcript expression in osteogenesis-induced hDPSCs. X. strumarium aqueous extract is a suitable candidate for bone repair because it promotes osteogenic differentiation in hDPSCs. Therefore this could be explored further in the treatment of bone disorders.


Differential effects of the extracellular microenvironment on human embryonic stem cell differentiation into keratinocytes and their subsequent replicative life span.
Culture microenvironment plays a critical role in the propagation and differentiation of human embryonic stem cells (hESCs) and their differentiated progenies. Although high efficiency of hESC differentiation to keratinocytes (hESC-Kert) has been achieved, little is known regarding the effects of early culture microenvironment and pertinent extracellular matrix (ECM) interactions during epidermal commitment on subsequent proliferative capacity of hESC-Kert. The aim of this study is to evaluate the effects of the different ECM microenvironments during hESC differentiation on subsequent replicative life span of hESC-Kert. In doing so, H1-hESCs were differentiated to keratinocytes (H1-Kert) in two differentiation systems. The first system employed autologous fibroblast feeder support, in which keratinocytes (H1-Kert(ACC)) were derived by coculture of hESCs with hESC-derived fibroblasts (H1-ebFs). The second system employed a novel decellularized matrix from H1-ebFs to create a dermoepidermal junction-like (DEJ) matrix. H1-Kert(AFF) were derived by differentiation of hESCs on the feeder-free system employing the DEJ matrix. Our study indicated that the feeder-free system with the use of DEJ matrix was more efficient in differentiation of hESCs toward epidermal progenitors. However, the feeder-free system was not sufficient to support the subsequent replicative capacity of differentiated keratinocytes. Of note, H1-Kert(AFF) showed limited replicative capacity with reduced telomere length and early cellular senescence. We further showed that the lack of cell-cell interactions during epidermal commitment led to heightened production of TGF-β1 by hESC-Kert during extended culture, which in turn was responsible for resulting in the limited replicative life span with cellular senescence of hESC-Kert derived under the feeder-free culture system. This study highlights for the first time the importance of the culture microenvironment and cell-ECM interactions during differentiation of hESCs on subsequent replicative life span and cellular senescence of the differentiated keratinocytes, with implications for use of these cells for applications in tissue engineering and regenerative medicine.


Infant mortality and low birth weight in cities of Northeastern and Southeastern Brazil.
To compare estimates of low birth weight (LBW), preterm birth, small for gestational age (SGA), and infant mortality in two birth cohorts in Brazil. The two cohorts were performed during the 1990s, in S o Lu s, located in a less developed area in Northeastern Brazil, and Ribeir o Preto, situated in a more developed region in Southeastern Brazil. Data from one-third of all live births in Ribeir o Preto in 1994 were collected (2,839 single deliveries). In S o Lu s, systematic sampling of deliveries stratified by maternity hospital was performed from 1997 to 1998 (2,439 single deliveries). The chi-squared (for categories and trends) and Student t tests were used in the statistical analyses. The LBW rate was lower in S o Lu s, thus presenting an epidemiological paradox. The preterm birth rates were similar, although expected to be higher in Ribeir o Preto because of the direct relationship between preterm birth and LBW. Dissociation between LBW and infant mortality was observed, since S o Lu s showed a lower LBW rate and higher infant mortality, while the opposite occurred in Ribeir o Preto. Higher prevalence of maternal smoking and better access to and quality of perinatal care, thereby leading to earlier medical interventions (cesarean section and induced preterm births) that resulted in more low weight live births than stillbirths in Ribeir o Preto, may explain these paradoxes. The ecological dissociation observed between LBW and infant mortality indicates that the LBW rate should no longer be systematically considered as an indicator of social development.


Mössbauer spectroscopy: recent developments.
In this review I have emphasized two basic points. First, Mössbauer spectroscopy has been widely used as a tool for basic research in solid-state physics, chemical structure, and magnetism. Second, the technique has been applied to many analytical and materials science problems where the nature of the sample prevents the use of more traditional approaches. Mössbauer spectroscopy research on frozen solutions, colloids, interface chemistry, and crystallographic transformations can be expected to increase rapidly in the next few years.


A non-randomised trial investigating the cost-effectiveness of Midwifery Group Practice compared with standard maternity care arrangements in one Australian hospital.
to compare cost-effectiveness of two models of maternity service delivery: Midwifery Group Practice (MGP) at a birth centre and standard care (SC). a prospective non-randomised trial. an Australian metropolitan hospital. women at 36 weeks gestation were approached in the birth centre or hospital antenatal clinics between March and December 2008. Of 170 consecutive women who met birth centre eligibility criteria, 70% (n=119) were recruited to the study. Women (MGP n=52 or standard care n=50) were followed through to 6 weeks postpartum. Publically funded care costs were collected from women's diaries, handheld pregnancy health records, medical records and the hospital accounting system. health-care costs to the hospital and government. generalised linear models with covariates of age, nulliparity, private health insurance (yes/no) and household income category. women receiving MGP care were less likely to experience induction of labour, required fewer antenatal visits, received more postnatal care, and neonates were less likely to be admitted to special care nursery than those receiving standard care. Statistically significant lower costs were found for women and babies receiving MGP care compared with women receiving standard care during pregnancy, labour and birth and postpartum to 6 weeks. MGP resulted in lower costs for the hospital ($AUD4,696 vs. $AUD5,521 p<0.001) and the government ($AUD4,722 vs. $AUD5,641 p<0.001). When baby costs were excluded MGP care remained statistically significantly cheaper than standard care. for women at low-risk of birth complications, Midwifery Group Practice was cost effective, and women experienced fewer obstetric interventions compared with standard maternity care. The evidence suggests Midwifery Group Practice is safe and economically viable.


Emergency department patients' perception of nurse caring behaviors.
To identify which behaviors performed by emergency department nurses were perceived by patients as important indicators of caring. Descriptive. Two private urban emergency departments. Ambulatory patients treated in the emergency department and interviewed by telephone within 30 days of discharge. The resulting sample consisted of a total of 288 interviews including 81 patients in the emergent group, 99 in the urgent group, and 108 in the nonurgent group. Of the 288 patients, 49% were male and 51% were female. Individual and composite measures of perceptions of nurse caring behaviors as measured with the Caring Behaviors Assessment, satisfaction with care, and patients' evaluation of their medical condition. Patients in all triage categories were found to assign the greatest importance to the technical nursing behaviors as indicators of nurse caring. Polychotomous logistic regression indicated that, although subscale differences occurred, they did not account for substantial differences among triage categories. Kruskall-Wallis one way ANOVA revealed no significant differences between triage levels and ratings of level of personal concern. Chi-square analysis indicated that patients in the emergent group identified fewer caring behaviors that the nurse must perform to demonstrate caring compared with patients in the nonurgent group. Patients experience nurse caring behavior most consistently from the technical aspects of nursing care.


Novel CNS malformations and skeletal anomalies in a patient with Beaulieu-boycott-Innes syndrome.
THO/TREX (transcription/export) is a conserved eukaryotic complex that plays a crucial role in gene expression and prevents DNA damage during mitosis and meiosis. In mammals, TREX is essential during embryogenesis, determining stem cell fate specification by regulating posttranscriptional self-renewal and differentiation in several tissues. It is composed of a core called THO, consisting of THOC1, 2, 5, 6, 7, and additional proteins. Bi-allelic mutations in THOC6 have been associated to Beaulieu-Boycott-Innes syndrome (BBIS), a syndromic form of intellectual disability (ID). To date, nine patients harbouring homozygous or compound heterozygous mutations in THOC6 have been reported. Despite the clinical heterogenity and subtle dysmorphic features in some individuals, distinctive facial features are tall forehead, short and upslanting palpebral fissures, deep set eyes, flat philtrum, and malocclusion. Nonlife threatening congenital anomalies are common, including cardiac and renal malformations, anteriorly displaced anus, cryptorchidism in males, submucous cleft palate, and corpus callosum dysgenesis. Affected patients usually have short stature, mild microcephaly, and mild to moderate ID. Here, we describe an Italian patient with BBIS, carrying two compound heterozygous loss-of-function (LoF) variants in THOC6 (c.577C > T, p.R193* and c.792_793delCA, p.V264Vfs*48). In addition to the common phenotype, she displays cerebellar hypoplasia with severe vermian dysgenesis and hydrocephalus due to aqueductal stenosis, multiple skeletal anomalies and hypergonadotropic hypogonadism. Thus, we review the previous cases and discuss the phenotypic spectrum of BBIS, providing further evidence regarding the pivotal role of TREX complex in human development.


The Nanoscale Wetting Parameter and Its Role in Interfacial Phenomena: Phase Transitions in Nanopores.
Through analysis of the statistical mechanical equations for a thin adsorbed film (gas, liquid, or solid) on a solid substrate or confined within a pore, it is possible to express the equilibrium thermodynamic properties of the film as a function of just two dimensionless parameters: a nanoscale wetting parameter, αw, and pore width, H*. The wetting parameter, αw, is defined in terms of molecular parameters for the adsorbed film and substrate and so is applicable at the nanoscale and for films of any phase. The main assumptions in the treatment are that (a) the substrate structure is not significantly affected by the adsorbed layer and (b) the diameter of the adsorbate molecules is not very small compared to the spacing of atoms in the solid substrate. We show that different surface geometries of the substrate (e.g., slit, cylindrical, and spherical pores) and various models of wall heterogeneity can be accounted for through a well-defined correction to the wetting parameter; no new dimensionless variables are introduced. Experimental measurements are reported for contact angles for various liquids on several planar substrates and are shown to be closely correlated with the nanoscale wetting parameter. We apply this approach to phase separation in nanopores of various geometries. Molecular simulation results for the phase diagram in confinement, obtained by the flat histogram Monte Carlo method, are reported and are shown to be closely similar to experimental results for capillary condensation, melting, and the triple point. The value of the wetting parameter, αw, is shown to determine the qualitative behavior (e.g., increase vs decrease in the melting temperature, capillary condensation vs evaporation), whereas the pore width determines the magnitude of the confinement effect. The triple point temperature and pressure for the confined phase are always lower than those for the bulk phase for all cases studied.


Water residence time (age) and flow path exert synchronous effects on annual characteristics of dissolved organic carbon in terrestrial runoff.
Catchment hydro-physical controls on the interannual variability of dissolved organic carbon (DOC) in terrestrial watershed runoff, important for water quality, ecosystem structure, and foodweb dynamics, are not well understood. To address this, we simulated water residence time ("age") and flow path of terrestrial runoff and analyzed their mediating effect on relationships between annual runoff volume, DOC concentration, and DOC age. We applied this analysis to a snow-influenced watershed in California's Sierra Nevada (USA) across a range of soil types, elevations (90-4210 m), and years (1950-1999). Simulated increases in annual runoff volume were accompanied by younger ages (r2 = 0.53-0.63) of DOC in quickflow, comprised of surface runoff and lateral flow through soil. Increases in annual runoff volume were also accompanied by gentler relationships between intra-annual (weekly) values of DOC concentration and runoff volume, regression-slopes of which followed a power-law relationship to annual runoff (r2 = 0.12-0.92) for approximately 70% of the watershed. Simulations including dynamics of water age and soil temperature produced annual ages of quickflow DOC ranging from 1 to 70 days over all soil types and water years. Similarity of this range to an observed, 1-69 day range in half-lives of relatively labile DOC in previous studies suggests substantial interannual and spatial variability in the biodegradability of DOC in terrestrial runoff. Simulations excluding dynamics of water age and soil temperature predicted order-of-magnitude less interannual variability in age of quickflow DOC, demonstrating the important effect of interannual variability in soil-water interaction times. These findings suggest that the distribution of DOC bioprocessing along transitions between terrestrial and aquatic systems may be strongly influenced by year-to-year variability in age of water.


UNCOMMON SIDE EFFECTS OF SUNITINIB THERAPY IN A PATIENT WITH METASTATIC RENAL CELL CANCER: CASE REPORT.
- Sunitinib is an orally administered multikinase inhibitor. This therapy can provoke uncommon side effects such as pancytopenia, tumor lysis syndrome, cardiac disorders, thromboembolic incidents, intestinal perforation, pancreatitis, acute renal failure, etc. We report a case of a 63-year-old female admitted to the hospital due to abdominal pain, nausea, vomiting and elevated blood pressure. One month earlier, sunitinib therapy for metastatic renal cell carcinoma was initiated. During the first cycle of therapy, after three weeks of sunitinib 50 mg daily, symptoms started and she stopped taking the drug. At admission, laboratory tests revealed elevated serum and urine amylase, C-reactive protein, urea and creatinine, and lowered platelet and leukocyte counts and hemoglobin value. Urine test showed proteinuria, erythrocyturia, leukocyturia and granulated cylinder. The patient was diagnosed with acute pancreatitis grade III, acute renal failure grade II, pancytopenia and urinary infection, and was hospitalized for five days. She was treated symptomatically and with antibiotic therapy because of persistently elevated C-reactive protein and pathologic urinary sediment, which led to subjective and clinical improvement. Acute pancreatitis, renal insufficiency and pancytopenia are rarely described side effects of sunitinib therapy, and clear connection between these conditions and drug activity is not yet determined. Medical specialists who prescribe and treat patients with sunitinib should be aware of the possible occurrence of these conditions and perform regular checkups of sunitinib treated patients.


Transport of the influenza virus genome from nucleus to nucleus.
The segmented genome of an influenza virus is encapsidated into ribonucleoprotein complexes (RNPs). Unusually among RNA viruses, influenza viruses replicate in the nucleus of an infected cell, and their RNPs must therefore recruit host factors to ensure transport across a number of cellular compartments during the course of an infection. Recent studies have shed new light on many of these processes, including the regulation of nuclear export, genome packaging, mechanisms of virion assembly and viral entry and, in particular, the identification of Rab11 on recycling endosomes as a key mediator of RNP transport and genome assembly. This review uses these recent gains in understanding to describe in detail the journey of an influenza A virus RNP from its synthesis in the nucleus through to its entry into the nucleus of a new host cell.


Fabrication of well-defined mushroom-shaped structures for biomimetic dry adhesive by conventional photolithography and molding.
Biomimetic dry adhesives have many attractive features, such as reversible and repeatable adhesion against various surfaces. This paper presents a method for the simple fabrication of biomimetic dry adhesives composed of a mushroom-shaped structure, which is based on conventional photolithography and molding. Firstly a masked and a maskless exposure are performed on the top and bottom of a photoresist, respectively, that generates microholes with an undercut after development. This structured photoresist is then used for molding, leading to mushroom-shaped structural features after sacrificing the photoresist. Because of the convenience of photolithography, the proposed method has the potential to fabricate various dry adhesives cost-efficiently.


Influence of drying protocol with isopropyl alcohol on the bond strength of resin-based sealers to the root dentin.
This study compared the bond strength, interfacial ultrastructure, and tag penetration of resin-based sealers applied to smear-free radicular dentin using 70% isopropyl alcohol as the active final rinse. Eighty root canals were prepared and assigned to 2 groups (n = 40) according to the drying protocol: paper points or 70% isopropyl alcohol. Then, roots were divided into 4 subgroups (n = 10) with respect to the sealer and obturation material: AH Plus (Dentsply De Trey GmbH, Konstanz, Germany) and gutta-percha (AH/GP), Hybrid Root SEAL (Sun Medical, Tokyo, Japan) and gutta-percha (HR/GP), Epiphany SE (Pentron Clinical Technologies, Wallingford, CT) and gutta-percha (EP/GP), and Epiphany SE and Resilon (EP/RS). Roots were sectioned, and the push-out test was performed. Failure modes were examined under stereomicroscopy and sealer penetration into the dentinal tubules under scanning electron microscopy. Data were statistically analyzed by 2-way analysis of variance post hoc Tukey tests with a significant level of 5%. Overall, canals dried with isopropyl alcohol showed significantly higher bond strength values (2.11 ± 1.74 MPa) than with paper points (1.81 ± 1.73 MPa) (P < .05). The HR/GP group showed lower bond strength than the AH/GP group (P < .05) but higher than the EP/GP and EP/RS groups (P < .05). The most frequent type of failure was cohesive in the AH/GP and HR/GP groups and adhesive in the EP/GP and EP/RS groups. Scanning electron microscopic evaluation revealed better adaptation of the adhesive interface in the AH/GP and HR/GP groups in comparison with the EP/GP and EP/RS groups. A final rinse with EDTA and 70% isopropyl alcohol improved the bond strength and penetration of the sealers into dentinal tubules of the root.


Targeted gene expression profiling in Leishmania braziliensis and Leishmania guyanensis parasites isolated from Brazilian patients with different antimonial treatment outcomes.
In Brazil, cutaneous leishmaniasis represents a serious public health problem, and chemotherapy is an important element of the clinical management of this disease. However, treatment efficacy is variable, a phenomenon that might be due to host and parasite (e.g., drug resistance) factors. To better understand the possible contribution of parasite factors to this phenomenon, we characterised 12 Leishmania braziliensis (LB) and 25 Leishmania guyanensis (LG) isolates collected from patients experiencing different antimonial treatment outcomes. For each isolate, promastigote cultures were grown in duplicate and were harvested at the late-log and stationary phases of growth. The RNA expression profiles of six genes encoding proteins with roles in antimony metabolism (AQP1, MRPA, GSH1, GSH2, TRYR and TDR1) were assessed by means of real-time quantitative PCR. Molecular data were compared to the clinical phenotypes. Within LB, we did not find statistically significant differences in the expression levels of the examined genes among isolates from patients with different treatment outcomes. In LG, GSH1 (encoding gamma-glutamylcysteine synthetase, gamma-GCS) was overexpressed in therapeutic failure isolates regardless of the growth curve phase. This finding reveals the predictive potential of promastigote expression curves for the prognosis of cutaneous leishmaniasis caused by LG in Brazil.


Asthma control: do patients' and physicians' opinions fit in with patients' asthma control status?
The aim of this study was to determine the level of asthma control and the concordance between physicians' and patients' opinions and the real patients' situation. A total of 777 subjects (55% female) with asthma were recruited. The study comprised a 4-week follow-up period, during which subjects completed a diary recording peak expiratory flow (PEF), symptoms, and use of rescue medication. At the end, both physicians and patients rated asthma control status. The level of control was evaluated using a composite measure. Agreement among subjective assessments of asthma control by patients and physicians and objective evaluation with the composite measure was assessed through kappa scores. A total of 518 (67%) patients had "not well-controlled asthma," being the remaining "totally controlled" (8%) or "well-controlled" (25%). "Not well-controlled asthma" was more frequent in smokers (82%) than in ex-smokers (70%) or in non-smokers (62%; p=0.0002). Kappa scores between patients' and physicians' opinions and the real patients' status were 0.02 (95% CI, 0.01-0.05) and 0.07 (95% CI, 0.03-0.09), respectively. In conclusion, current level of asthma control is suboptimal for a majority (67%) of patients in Spain. Besides, asthma control is worse in smokers. Neither patients' nor physicians' opinions agree with patients' real situation. Both patients and physicians accept as normal a suboptimal status of their disease.


Determinants of survival in myelofibrosis patients undergoing allogeneic hematopoietic cell transplantation.
We aimed to evaluate the determinants of survival in myelofibrosis patients undergoing allogeneic hematopoietic cell transplantation (allo-HCT) and to describe factors predicting the main post-HCT complications. This retrospective study by the European Society for Blood and Marrow Transplantation included 2916 myelofibrosis patients who underwent first allo-HCT from an HLA-identical sibling or unrelated donor between 2000 and 2016. After a median follow-up of 4.7 years from transplant, projected median survival of the series was 5.3 years. Factors independently associated with increased mortality were age ≥ 60 years and Karnofsky Performance Status <90% at transplant, and occurrence of graft failure, grades III-IV acute graft-vs.-host disease (aGVHD), and disease progression/relapse during follow-up. The opposing effects of chronic graft-vs.-host disease (GVHD) on non-relapse mortality and relapse incidence resulted in a neutral influence on survival. Graft failure increased in unrelated donor recipients and decreased with myeloablative conditioning (MAC) and negative donor/recipient cytomegalovirus serostatus. Risk of grades III-IV aGVHD was higher with unrelated donors and decreased with MAC. Relapse incidence tended to be higher in patients with intermediate-2/high-risk DIPSS categories and to decrease in CALR-mutated patients. Acute and chronic GVHD reduced the subsequent risk of relapse. This information has potential implications for patient counseling and clinical decision-making.


[Cajal bodies in nuclei of oocytes from the scorpion fly Panorpa communis].
The absence of nucleoli and rRNA synthesis in oocyte nuclei of the scorpion fly, Panorpa communis, was demonstrated using hybridization in situ. The immunocytochemical characteristics of a single large nuclear body of P. communis oocytes has been done which enabled us to recognize this structure as a Cajal body (CB). A marker of CBs coilin was detected in CBs of P. communis oocytes in addition to Sm-epitope of small nuclear RNPs (snRNPs), trimethylguanosine cap of snRNAs, and non-snRNP splicing factor, SC35. Besides, inactive hyperphosphorylated and nonphosphorylated forms of RNA polymerase II were also found in CBs. We put forward a suposition on possible roles of CBs in P. communis oocytes. On the one hand, CBs take part in the storage of RNA transcription and processing components to be used later during embryogenesis. On the other hand, they provide the primary assembly and redistribution of pre-mRNA transcription and processing components of the oocyte itself.


The decline in pulsatile GH secretion throughout early adulthood in mice is exacerbated by dietary-induced weight gain.
The transition between puberty and adulthood is accompanied by a slowing in linear growth. Although GH is a key factor that drives somatic development into adulthood, early adulthood coincides with a reduction in circulating levels of GH. To this extent, a pathological decline in postpubertal GH secretion is detrimental to attainment of peak lean muscle mass and bone mass and promotes adiposity and increases susceptibility to the development of obesity in adulthood. Here we characterized pulsatile GH secretion in C57BL/6J mice at 12 and 16 wk of age. Deconvolution analysis of these measures reveals a reduction in pulsatile GH secretion between 12 and 16 wk of age. Dietary intervention with high-fat feeding at 8 wk of age results in a significant increase in adiposity, the development of glucose intolerance, and hyperinsulinemia. We show the exacerbation of the age-associated decline in pulsatile GH secretion in high-fat-fed mice after 4 wk of dietary intervention (at 12 wk of age), and a further suppression of pulsatile GH secretion by 8 wk of dietary intervention (at 16 wk of age). Suppressed pulsatile secretion of GH did not coincide with an elevation in circulating free fatty acids. Rather, we observed increased hepatic triglyceride content and an eventual decrease in circulating levels of IGF-I. Given the established role of GH in maintaining healthy aging, we anticipate that an advancing of the age-associated decline in pulsatile GH secretion as a consequence of dietary-induced weight gain may have long-term ramifications on adult health.


Design rules for glass formation from model molecules designed by a neural-network-biased genetic algorithm.
The glass transition - an apparent amorphous solidification process - is a central feature of the physical properties of soft materials such as polymers and colloids. A key element of this phenomenon is the observation of a broad spectrum of deviations from an Arrhenius temperature of dynamics in glass-forming liquids, with the extent of deviation quantified by the "fragility" of glass formation. The underlying origin of "fragile" glass formation and its dependence on molecular structure remain major open questions in condensed matter physics and soft materials science. Here we employ molecular dynamics simulations, together with a neural-network-biased genetic algorithm, to design and study model rigid molecules spanning a broad range of fragilities of glass formation. Results indicate that fragility of glass formation can be controlled by tuning molecular asphericity, with extended molecules tending to exhibit low fragilities and compact molecules tending toward higher fragilities. The glass transition temperature itself, on the other hand, correlates well with high-temperature activation behavior and with density. These results point the way towards rational design of glass-forming liquids spanning a range of dynamical behavior, both via these physical insights and via future extensions of this evolutionary design strategy to real chemistries. Finally, we show that results compare well with predictions of the nonlinear Langevin theory of liquid dynamics, which is a precursor of the more recently developed elastically collective nonlinear Langevin equation theory of Mirigian and Schweizer, identifying this framework as a promising basis for molecular design of the glass transition.


Crossed Corticostriatal Projections in the Macaque Brain.
In nonhuman primates, major input to the striatum originates from ipsilateral cortex and thalamus. The striatum is a target also of crossed corticostriatal (CSt) projections from the contralateral hemisphere, which have been so far somewhat neglected. In the present study, based on neural tracer injections in different parts of the striatum in macaques of either sex, we analyzed and compared qualitatively and quantitatively the distribution of labeled CSt cells in the two hemispheres. The results showed that crossed CSt projections to the caudate and the putamen can be relatively robust (up to 30% of total labeled cells). The origin of the direct and the crossed CSt projections was not symmetrical as the crossed ones originated almost exclusively from motor, prefrontal, and cingulate areas and not from parietal and temporal areas. Furthermore, there were several cases in which the contribution of contralateral areas tended to equal that of the ipsilateral ones. The present study is the first detailed description of this anatomic pathway of the macaque brain and provides the substrate for bilateral distribution of motor, motivational, and cognitive signals for reinforcement learning and selection of actions or action sequences, and for learning compensatory motor strategies after cortical stroke.SIGNIFICANCE STATEMENT In nonhuman primates the striatum is a target of projections originating from the contralateral hemisphere (crossed CSt projections), which have been so far poorly investigated. The present study analyzed qualitatively and quantitatively in the macaque brain the origin of the crossed CSt projections compared with those originating from the ipsilateral hemisphere. The results showed that crossed CSt projections originate mostly from frontal and rostral cingulate areas and in some cases their contribution tended to equal that from ipsilateral areas. These projections could provide the substrate for bilateral distribution of motor, motivational, and cognitive signals for reinforcement learning and action selection, and for learning compensatory motor strategies after cortical stroke.


[Necrotizing cutaneous vasculitis with massive gastrointestinal bleeding following naproxen treatment].
Cutaneous necrotizing vasculitis is usually induced by an acute infection or exposure to a drug. Cutaneous vasculitis may precede severe systemic involvement, and may end in death. Accordingly, diagnosis of cutaneous vasculitis, identification of etiological factors, follow-up for systemic involvement and treatment are important. The authors present a case study of a 58-year-old male with fever and extensive eruption involving the trunk and extremities which appeared two days after initiation of treatment with oral naproxen. The clinical and histological findings were consistent with cutaneous necrotizing leukocytoclastic vasculitis. Two events of massive upper gastrointestinal bleeding occurred during treatment with systemic corticosteroids. The possible etiological factors and the mechanisms involved in the induction of the vasculitis and the gastrointestinal bleeding are discussed.


[Clinical features of neuroses in left-handed children].
A study covering 100 sinistral and 100 dextral neurotic children revealed a higher percentage of persons with manifestations of motor disinhibition, phobias and anxiety among sinistral children. It was also in this group that a considerable number of children presented at the beginning of schooling such phenomena as "mirror" writing and attempts of reading and counting from right to left. Elements of "mirror" thinking seemed to explain difficulties in learning writing and reading observed in such children. The authors discuss the questions of prophylactic measures in sinistral children with neuroses.


Whole cell melanoma vaccine genetically modified to stem cells like phenotype generates specific immune responses to ALDH1A1 and long-term survival in advanced melanoma patients.
Allogeneic whole cell gene modified therapeutic melanoma vaccine (AGI-101H) comprising of two melanoma cell lines transduced with cDNA encoding fusion protein composed of IL-6 linked with the soluble IL-6 receptor (sIL-6R), referred to as H6 was developed. H6 served as a molecular adjuvant, however, it has altered vaccine cells phenotype towards melanoma stem cells (MSC)-like with high activity of aldehyde dehydrogenase isoenzyme (ALDH1A1). AGI-101H was applied in advanced melanoma patients with non-resected and resected disease. In the adjuvant setting, it was combined with surgery in case of recurring metastases, which were surgically removed and vaccination continued. A significant fraction of AGI-101H treated melanoma patients is still alive (11-19 years). Out of 106 living patients, 39 were HLA-A2 positive and were the subject of the study. Immunization of melanoma patients resulted in the generation of cytotoxic CD8+ T cells specific for ALDH1A1, which were detected in circulation by HLA-A0201 MHC dextramers loaded with ALDH1A188-96(LLYKLADLI) peptide. Phenotypically they were central memory CD8+ T cells. Re-stimulation with ALDH1A188-96ex vivo resulted in IFN-γ secretion and cells degranulation. Following each vaccine dose administration, the number of ALDH1A1-CD8+ T cells increased in circulation and returned to the previous level until next dose injection (one month). ALDH1A1-CD8+ T cells were also found, however in the lower number than in vaccinated patients, in the circulation of untreated melanoma with stage IV but were not found in stage II or III and healthy donors. Specific anti-ALDH1 antibodies were present in treated patients. Long-term survival suggests immuno-targeting of MSC in treated patients.


Contagious behavior: an alternative approach to mirror-like phenomena.
Contagious behaviors such as yawning and itching/scratching have mirror-like properties and clearly defined stimulus and motor parameters; they are also relatively easy to study and should be part of the debate about mirror neurons and the neurological mechanisms of social behavior. The broadly tuned, multimodal stimuli of contagious behavior challenge present accounts of mirror mechanisms that focus on specific, mirrored acts.


Cluster of an Unusual Amnestic Syndrome - Massachusetts, 2012-2016.
In November 2015, a neurologist in the Boston, Massachusetts, area reported four cases of an uncommon amnestic syndrome involving acute and complete ischemia of both hippocampi, as identified by magnetic resonance imaging (MRI), to the Massachusetts Department of Public Health (MDPH) (1). A subsequent e-mail alert, generated by the Massachusetts Board of Registration in Medicine and sent to relevant medical specialists (including neurologists, neuroradiologists, and emergency physicians), resulted in the identification of 10 additional cases that had occurred during 2012-2016. All 14 patients (mean and median age = 35 years) had been evaluated at hospitals in eastern Massachusetts. Thirteen of the 14 patients underwent routine clinical toxicology screening at the time of initial evaluation; eight tested positive for opioids, two for cocaine, and two for benzodiazepines. Apart from sporadic cases (2-6), this combination of clinical and imaging findings has been reported rarely. The apparent temporospatial clustering, relatively young age at onset (19-52 years), and associated substance use among these patients should stimulate further case identification to determine whether these observations represent an emerging syndrome related to substance use or other causes (e.g., a toxic exposure).


Ebbie: automated analysis and storage of small RNA cloning data using a dynamic web server.
DNA sequencing is used ubiquitously: from deciphering genomes to determining the primary sequence of small RNAs (smRNAs). The cloning of smRNAs is currently the most conventional method to determine the actual sequence of these important regulators of gene expression. Typical smRNA cloning projects involve the sequencing of hundreds to thousands of smRNA clones that are delimited at their 5' and 3' ends by fixed sequence regions. These primers result from the biochemical protocol used to isolate and convert the smRNA into clonable PCR products. Recently we completed a smRNA cloning project involving tobacco plants, where analysis was required for approximately 700 smRNA sequences. Finding no easily accessible research tool to enter and analyze smRNA sequences we developed Ebbie to assist us with our study. Ebbie is a semi-automated smRNA cloning data processing algorithm, which initially searches for any substring within a DNA sequencing text file, which is flanked by two constant strings. The substring, also termed smRNA or insert, is stored in a MySQL and BlastN database. These inserts are then compared using BlastN to locally installed databases allowing the rapid comparison of the insert to both the growing smRNA database and to other static sequence databases. Our laboratory used Ebbie to analyze scores of DNA sequencing data originating from an smRNA cloning project. Through its built-in instant analysis of all inserts using BlastN, we were able to quickly identify 33 groups of smRNAs from approximately 700 database entries. This clustering allowed the easy identification of novel and highly expressed clusters of smRNAs. Ebbie is available under GNU GPL and currently implemented on http://bioinformatics.org/ebbie/. Ebbie was designed for medium sized smRNA cloning projects with about 1,000 database entries. Ebbie can be used for any type of sequence analysis where two constant primer regions flank a sequence of interest. The reliable storage of inserts, and their annotation in a MySQL database, BlastN comparison of new inserts to dynamic and static databases make it a powerful new tool in any laboratory using DNA sequencing. Ebbie also prevents manual mistakes during the excision process and speeds up annotation and data-entry. Once the server is installed locally, its access can be restricted to protect sensitive new DNA sequencing data. Ebbie was primarily designed for smRNA cloning projects, but can be applied to a variety of RNA and DNA cloning projects.


Patients with isolated trisomy 8 in acute myeloid leukemia are not cured with cytarabine-based chemotherapy: results from Cancer and Leukemia Group B 8461.
To date, neither the clinical significance of isolated trisomy 8, the most frequent trisomy in acute myeloid leukemia (AML), nor the effect of age within a single cytogenetic group has been examined. We report a large cohort of adult trisomy 8 patients and examine whether increasing age within a homogeneous cytogenetic group alters clinical outcome. Characteristics and outcome of patients with isolated trisomy 8 enrolled in the prospective Cancer and Leukemia Group B (CALGB) cytogenetic study CALGB 8461 are described. Isolated trisomy 8 was identified in 42 (3.03%) of 1387 patients enrolled in five CALGB treatment protocols. These patients had a median age of 64 (range, 16-79) years, 50% female proportion, and a low frequency of hepatomegaly (10%) or splenomegaly (10%). Laboratory features included a median white blood count of 7.3 x 10(9)/L, nonspecific French-American-British distribution, with 36% of patients having Auer rods. Treatment outcome was unsatisfactory with a complete remission (CR) rate of 59%, median CR duration of 13.6 months, and median survival of 13.1 months. Older age adversely affected outcome; trisomy 8 patients > or =60 years had both an inferior CR rate (40% versus 88%; P = 0.004) and overall survival (median, 4.8 versus 17.5 months; P = 0.01), as compared with those <60 years of age. Of the patients <60 years of age, only four remain alive, and all received noncytarabine-based intensive chemotherapy, followed in three cases by autologous (n = 2) or allogeneic (n = 1) stem cell transplant in CR1. Adults with AML and isolated trisomy 8 have a poor outcome that is accentuated by increasing age and is rarely cured with cytarabine-based therapy. Alternative investigational treatments should be considered for individuals with this AML subset.


Generalized Anxiety Disorder 7-Item (GAD-7) Scores in Medically Authorized Cannabis Patients-Ontario and Alberta, Canada.
Despite increasing rates of legalization of medical cannabis worldwide, the current evidence available on its effect on mental health outcomes including anxiety is of mixed results. This study assesses the effect of medical cannabis on generalized anxiety disorder 7-item (GAD-7) scores in adult patients between 2014 and 2019 in Ontario and Alberta, Canada. An observational cohort study of adults authorized to use medical cannabis. The GAD-7 was administered at the time of the first visit to the clinic and subsequently over the follow-up time period of up to 3.2 years. Overall changes in GAD-7 scores were computed (mean change) and categorized as: no change (<1 point); improvement; or worsening-over time. A total of 37,303 patients had initial GAD-7 scores recorded and 5,075 (13.6%) patients had subsequent GAD-7 follow-up scores. The average age was 54.2 years (SD 15.7 years), 46.0% were male, and 45.6% noted anxiety symptoms at the baseline. Average GAD-7 scores were 9.11 (SD 6.6) at the baseline and after an average of 282 days of follow-up (SD 264) the average final GAD-7 score recorded was 9.04 (SD 6.6): mean change -0.23 (95% CI, -0.28 to -0.17, t[5,074]: -8.19, p-value <0.001). A total of 4,607 patients (90.8%) had no change in GAD-7 score from their initial to final follow-up, 188 (3.7%) had a clinically significant decrease, and 64 (1.3%) noted a clinically significant increase in their GAD-7 scores. Overall, there was a statistically significant decrease in GAD-7 scores over time (in particular, in the 6-12-month period). However, this change did not meet the threshold to be considered clinically significant. Thus, we did not detect clinical improvements or detriment in GAD-7 scores in medically authorized cannabis patients. However, future well-controlled clinical trials are needed to fully examine risks or benefits associated with using medical cannabis to treat anxiety conditions.


Effects of ZnO nanoparticles in plants: Cytotoxicity, genotoxicity, deregulation of antioxidant defenses, and cell-cycle arrest.
Cytotoxicity, genotoxicity, and biochemical effects were evaluated in the plants Allium cepa, Nicotiana tabacum, and Vicia faba following exposure to ZnO nanoparticles (np; diameter, ∼85nm). In the root meristems of Allium cepa cells, we observed loss of membrane integrity, increased chromosome aberrations, micronucleus formation, DNA strand breaks, and cell-cycle arrest at the G2/M checkpoint. In Vicia faba and Nicotiana tabacum, we observed increased intracellular ROS production, lipid peroxidation, and activities of some antioxidant enzymes. TEM images revealed gross morphological alterations and internalization of the np. Our findings provide evidence of ZnO np toxicity, characterized by deregulation of components of ROS-antioxidant machinery, leading to DNA damage, cell-cycle arrest, and cell death. These plants, especially Allium cepa, are reliable systems for assessment of np toxicology.


Endothelial Progenitor-Cell-Derived Exosomes Induced by Astragaloside IV Accelerate Type I Diabetic-wound Healing via the PI3K/AKT/mTOR Pathway in Rats.
We explore the effects of endothelial progenitor cell (EPC)-derived exosomes (EPCexos) and of astragaloside IV (ASIV)-stimulated EPCexos (ASIV-EPCexos) on type I diabetic-wound healing, and determine the basic molecular mechanisms of action. EPCs were exposed to different concentrations of ASIV to generate ASIV-EPCexos. A chronic-wound healing model involving streptozotocin-stimulated diabetic rats was established. These rats were treated with EPCexos, ASIV-EPCexos, rapamycin, and wortmannin. Wound healing was evaluated by direct photographic observation, hematoxylin and eosin staining, and Masson's trichrome staining. ASIV treatment increased the abilities of EPCs (e.g., proliferation), as well as exosome secretion. EPCexo showed a "cup holder" like structure. Treatment with ASIV-EPCexos increased the wound-healing rate, collagen-deposition area, bromodeoxyuridine uptake, VEGF expression, and the number of CD31- and αSMA- positive cells, whereas decreased epidermal thickness and CD45 expression. The expression of the PI3K/AKT/mTOR pathway increased, whereas the expression of inflammatory factor decreased. However, rapamycin and wortmannin reversed these changes. ASIV-EPCexos may accelerate type I diabetic-wound healing via the PI3K/AKT/mTOR pathway. This study may lay the foundation for new clinical treatment options for patients with type I diabetic wounds.


Distinct pathways for NF-kappa B regulation are associated with aberrant macrophage IL-12 production in lupus- and diabetes-prone mouse strains.
One characteristic of mice prone to a variety of autoimmune diseases is the aberrant regulation of cytokine production by macrophages (Mphi), noted in cells isolated well before the onset of disease. Strikingly, the pattern of IL-12 dysregulation, in particular, is consistent with the nature of the autoimmune disease that will develop in each strain, i.e., elevated in mice prone to Th1-mediated organ-specific disease (nonobese diabetic (NOD) and SJL mice) and reduced in lupus-prone strains (MRL/+ and NZB/W). Mechanistically, the abnormal regulation of IL-12 in these strains was found to be strictly associated with novel patterns of Rel binding in vitro to the unique NF-kappaB site in the IL-12 p40 promoter. In this study, we report several new findings related to these Rel-kappaB interactions. Evaluation of the p40 NF-kappaB site in vivo, assessed by chromatin immunoprecipitation, revealed Rel usage patterns similar to those found in vitro using EMSA, with preferential association of the p40 kappaB site with c-Rel in NOD Mphi but with p50 in NZB/W Mphi. Moreover, blocking c-Rel in primary Mphi, using short interfering RNA, selectively blocked IL-12 production and normalized the minimal, residual IL-12 levels. Nuclear extracts from NOD Mphi were characterized by c-Rel hyperphosphorylation, and dephosphorylation of nuclear proteins completely blocked binding to the kappaB site. In contrast, elevated IkappaB appears to be a likely mechanism accounting for the reduced nuclear c-Rel levels noted in NZB/W Mphi. Alterations in NF-kappaB metabolism thus appear to define a pathway regulating intrinsic IL-12 defects in both diabetes- and lupus-prone strains.


[Publication languages of Impact Factor journals and of medical bibliographic databanks (reprint)].
A preference for English-language sources during determination of Journal Impact Factors (IF) was discussed, IF being published in the annual Journal Citation Reports (JCR). The JCR are derived from data in Science Citation Index (SCI). The aim of this study was, therefore, (i) to review publication countries and languages in JCR, (ii) publication languages in SCI in comparison to further recognised medical bibliographic databanks. Searching (i) countries and languages in JCR Science-Editions 1997 and 1998, (ii) language distributions in publication years 1995 - 2000 in bibliographic databanks SCI, MEDLINE (ME) and EMBASE (EM). (i) Almost 70 % journals in JCR 1997 and 1998 were published in USA, United Kingdom, or The Netherlands. Of two language options present, a number of English-classified journals contained > 90 % articles in other languages, whereas > 90 % publications in English could occur in Multi-Language (ML) journals, thereby complicating statistical comparisons. 83,9 % JCR-periodicals in 1997 and 85,6 % in 1998 were classified English. English/ML ratios increased exponentially with increasing IF. (ii) 95,5 % of the articles documented 1995 - 2000 in whole SCI and in our constructed SCI segment "Medicine and related areas" were written in English, compared to 88,5 % in ME and 89,8 % in EM. The SCI Medicine segment was 15 % more comprehensive than either MEDLINE or EMBASE. Highly significant differences of language distributions in SCI vs. MEDLINE and especially SCI vs. EMBASE were observed. Retrieval rates in SCI of German-, French-, Japanese- and Chinese-language medical papers published in 2000 were impressively augmented by EMBASE and MEDLINE. (i) Anglo-American publishers" countries and English-language journals prevail in JCR with respect to numbers and IF levels. Publication language English favours citation frequency. (ii) Of databanks studied, SCI shows a maximum preference for English-language sources, thereby causing an English Language Bias during IF derivation.


Junctional transfer in wounded cultures of bovine aortic endothelial cells.
Both in vivo and in vitro, endothelial cells form continuous monolayers displaying growth control by cell density as well as cell contact. Several functions of endothelium are dependent upon maintenance of an intact monolayer. When such a monolayer is injured, endothelial cells migrate out from the wound edges and proliferate to cover the area of denudation. The response of endothelium to wounding is therefore a matter of both growth control and cellular motility. Since intercellular gap junctions have been postulated to have a role in growth control, we set out to determine whether junctional communication was altered in the activated, migrating cells. We found extensive transfer of microinjected fluorescent dye molecules (Lucifer yellow CH) in cells at or near the wound edge at various times after wounding (1 minute to 48 hours). However, cells near a wound edge transferred dye at a frequency significantly diminished from that observed in undisturbed monolayers (81.1 +/- 1.5 (SE) versus 92.7 +/- 0.8% of adjacent cells received dye). There was a significant positive correlation between transfer frequency and distance from the wound (expressed as intervening cells) but not between transfer frequency and time after wounding. These results suggest that endothelial cells in a wounded monolayer retain junctional communication at a slightly but significantly reduced level and that increased motility and the wound healing response do not necessarily correlate with total loss of communication.


Genetics in diabetic retinopathy: current concepts and new insights.
There is emerging evidence which indicates the essential role of genetic factors in the development of diabetic retinopathy (DR). In this regard it should be highlighted that genetic factors account for 25-50% of the risk of developing DR. Therefore, the use of genetic analysis to identify those diabetic patients most prone to developing DR might be useful in designing a more individualized treatment. In this regard, there are three main research strategies: candidate gene studies, linkage studies and Genome-Wide Association Studies (GWAS). In the candidate gene approach, several genes encoding proteins closely related to DR development have been analyzed. The linkage studies analyze shared alleles among family members with DR under the assumption that these predispose to a more aggressive development of DR. Finally, Genome-Wide Association Studies (GWAS) are a new tool involving a massive evaluation of single nucleotide polymorphisms (SNP) in large samples. In this review the available information using these three methodologies is critically analyzed. A genetic approach in order to identify new candidates in the pathogenesis of DR would permit us to design more targeted therapeutic strategies in order to decrease this devastating complication of diabetes. Basic researchers, ophthalmologists, diabetologists and geneticists should work together in order to gain new insights into this issue.


Functional brain networks associated with cognitive control, cocaine dependence, and treatment outcome.
Individuals with cocaine dependence often evidence poor cognitive control. The purpose of this exploratory study was to investigate networks of functional connectivity underlying cognitive control in cocaine dependence and examine the relationship of the networks to the disorder and its treatment. Independent component analysis (ICA) was applied to fMRI data to investigate if regional activations underlying cognitive control processes operate in functional networks, and whether these networks relate to performance and treatment outcome measures in cocaine dependence. Twenty patients completed a Stroop task during fMRI prior to entering outpatient treatment and were compared to 20 control participants. ICA identified five distinct functional networks related to cognitive control interference events. Cocaine-dependent patients displayed differences in performance-related recruitment of three networks. Reduced involvement of a "top-down" fronto-cingular network contributing to conflict monitoring correlated with better treatment retention. Greater engagement of two "bottom-up" subcortical and ventral prefrontal networks related to cue-elicited motivational processing correlated with abstinence during treatment. The identification of subcortical networks linked to cocaine abstinence and cortical networks to treatment retention suggests that specific circuits may represent important, complementary targets in treatment development for cocaine dependence.


[Changes in Na+, K+-ATPase and acetylcholinesterase activity in red cell membranes after freezing-thawing].
The activity of Na+, K+ATPase (EC 184.108.40.206) and acetylcholinesterase (EC 18.104.22.168) as well as the content of masked and exposed SH-groups in sealed and unsealed erythrocyte ghosts were studied as affected by single rapid freezing-thawing. The freezing-thawing procedure resulted in different reactions of membrane-bound enzymes: Na+, K+-ATPase activity is doubled in sealed ghosts and unchanged in unsealed ones. In both types of ghosts the equal decrease in the AChE activity was found to be parallel with the diminution in the content of masked SH-groups but this cannot be referred to exposed SH-groups. The obtained results seem to suggest that the changes in the native conformation of membrane catalytic proteins resulted from cryodamage are responsible for the lowered aChE activity; the primary cause of increase Na+, K+-ATPase activity is due to the changes in the permeability and integrity of erythrocyte membrane, which are followed by the greater accessibility of the substrate (ATP) to the enzyme.


Two genes encode factors with NF-kappa B- and H2TF1-like DNA-binding properties.
NF-kappa B and H2TF1 are transcription factors that bind to related DNA sequence motifs. NF-kappa B is a constitutive factor in mature B lymphocytes and is inducible in immature B cells and nonlymphoid cells. H2TF1 is a constitutive ubiquitous transcription factor that activates expression of major histocompatibility complex class I genes. We report here the isolation of two zinc finger genes. One, named MBP-2 (major histocompatibility complex-binding protein 2), encodes a factor that has DNA-binding properties similar, if not identical, to the H2TF1 transcription factor. The second, named KBP-1 (kappa-enhancer binding protein 1), encodes a factor that is similar to, but distinct from, NF-kappa B in its DNA-binding properties and methylation interference pattern, suggesting that at least two proteins can bind to the immunoglobulin kappa gene enhancer.


How well are Indonesia's urban poor being provided access to quality reproductive health services?
Accommodating the needs of Indonesia's rapidly growing urban population is essential to reaching national reproductive health goals and international commitments. As in other rapidly urbanizing low- and middle-income countries, satisfying the needs of Indonesia's urban poor is both a high priority and a significant challenge. In this study, we assessed both how being from urban poor or near-poor households affects the quantity and quality of family planning and maternal health services received and the extent to which differentials had narrowed during the 2012-2017 period. This time interval is significant due to the introduction of a national social health insurance scheme in 2014, establishing the foundation for universal health care in the country. Data from the 2012 and 2017 Indonesian Demographic and Health Surveys were analyzed using logistic and multinomial logit regression. Poverty status was measured in terms of urban household wealth quintiles. For family planning, although urban poor and near-poor women made different method choices than non-poor women, no substantial 2017 differences in contraceptive prevalence, unmet need for family planning or informed choice were observed. However, urban poor women and to a lesser extent near-poor women systematically lagged non-poor urban women in both the quantity and quality of maternal health services received in connection with recent pregnancies. Significant maternal health service gains were observed for all urban women during the study reference period, with gains for poor and near poor urban women exceeding those for non-poor on several indicators. While the deployment of pro-poor interventions such as the national social health insurance scheme is likely to have contributed to these results, evidence suggesting that the scheme may not be influencing consumer health-seeking behaviors as had been anticipated along with continued limitations in public health sector supply-side readiness resulting in service quality issues suggest that more will have to be done.


The role of tumour necrosis factor-alpha (TNF-alpha) and platelet-activating factor (PAF) interaction on murine candidosis.
Tumour necrosis factor-alpha (TNF-alpha) is related to some other factors in addition to being the essential cytokine of the sepsis which results from Candida infections. In our study, we investigated serum TNF-alpha levels, measured by enzyme-linked immunosorbent assay (ELISA), and platelet-activating factor (PAF)-like activity, measured by high-pressure liquid chromatography (HPLC) of the mice infected with Candida species. The PAF antagonist, ginkgolide BN 52021 was used to evaluate the possible interaction between TNF-alpha and PAF. The average TNF-alpha levels were found to be 396, 489, 699 and 803 pg ml(-1) on the 4th, 5th, 6th and 19th days of Candida albicans infection, respectively (P<0.05). There was no statistically significant difference between the serum TNF-alpha levels of the groups infected with other Candida species, such as C. kefyr, C. krusei and C. tropicalis (P>0.05). Serum TNF-alpha levels were found to be more significantly different in mice with C. albicans infection that were injected with PAF antagonists on the 6th day (23 pg ml(-1)). It was therefore thought that PAF antagonists have an inhibitory effect on TNF-alpha production. No significant difference was found between PAF levels in the three groups: healthy control mice, C. albicans-infected mice and C. albicans-infected mice given PAF antagonists (466 milli-absorbance unit (mAU), 475 mAU and 329 mAU, respectively). It was noticed that the positive interaction between PAF and TNF-alpha was not important after the first 4 days of the infection had passed.


Common and unexpected findings in mummies from ancient Egypt and South America as revealed by CT.
Computed tomography (CT) has proved to be a valuable investigative tool for mummy research and is the method of choice for examining mummies. It allows for noninvasive insight, especially with virtual endoscopy, which reveals detailed information about the mummy's sex, age, constitution, injuries, health, and mummification techniques used. CT also supplies three-dimensional information about the scanned object. Mummification processes can be summarized as "artificial," when the procedure was performed on a body with the aim of preservation, or as "natural," when the body's natural environment resulted in preservation. The purpose of artificial mummification was to preserve that person's morphologic features by delaying or arresting the decay of the body. The ancient Egyptians are most famous for this. Their use of evisceration followed by desiccation with natron (a compound of sodium salts) to halt putrefaction and prevent rehydration was so effective that their embalmed bodies have survived for nearly 4500 years. First, the body was cleaned with a natron solution; then internal organs were removed through the cribriform plate and abdomen. The most important, and probably the most lengthy, phase was desiccation. After the body was dehydrated, the body cavities were rinsed and packed to restore the body's former shape. Finally, the body was wrapped. Animals were also mummified to provide food for the deceased, to accompany the deceased as pets, because they were seen as corporal manifestations of deities, and as votive offerings. Artificial mummification was performed on every continent, especially in South and Central America.


[Peripheral hepatic artery embolization: experimental study in dogs].
Peripheral hepatic artery embolization was induced in dogs by intra-arterial injection of cisplatin mixture. The result showed that development of hepatocellular damage and liver infarction was accompanied synchronously with impairment of the liver-function tests during 3-7 days after the embolization, and this impairment recovered gradually afterward. The emboli were 86 microns +/- 46 microns in diameter, and were mainly found in the hepatic micro-arteries. The presence of peripheral hepatic artery occlusion was considered effective in preventing the development of collateral circulation.


Why is your patient sedated?
The purpose of this paper is to examine the recording of clinical indication for prescribed sedative/hypnotic (SH) medications in a large, acute tertiary referral hospital. All hospital inpatients' medications (n = 367) were audited for prescription details regarding SH medications. Medical notes were then examined for evidence of a recorded indication for such medications. SH medications were prescribed to many hospital inpatients (42.5 per cent) during admission. An indication was documented in the nursing or medical records for 24.4 per cent of patients who were prescribed such medications. Nurses rather than by doctors prescribed most SH medications (74 vs 26 per cent, respectively, p = 0.003). Some patients receiving SH medications were both over 65 and impaired in their mobility (19.2 per cent). The treatment indication was documented in 47 per cent. Most patients prescribed SH medications have nothing in their medical record explaining why these drugs are being used, including half of the elderly, less mobile patients. All health professionals dealing with SH medications and doctors in particular need to justify the use of such medications in the medical record. For the particularly high-risk groups where SH medications are potentially more dangerous, explicit guidance on why and how such medications are to be used must be provided by prescribers. For the first time, data are presented on documentation rates for clinical indication of prescribed SH medications across a large acute hospital, and highlights significant shortcomings in practice. This study should inform other organisations of the need to be mindful of facilitating greater compliance with good prescribing practice.


Pharmacological strategies to prevent macrovascular disease in NIDDM.
The pathogenesis of atherosclerosis and vascular thrombosis in NIDDM is reviewed. Evidence that suggests a role for chronic hyperglycemia, in association with other vascular risk markers, is presented. Based on this framework, a multifactorial approach to the prevention of progression of macrovascular disease in NIDDM is discussed. Results from a recent consensus conference sponsored by the American Diabetes Association regarding approaches to glycemic regulation in people with NIDDM are reviewed. It is concluded that preventive approaches will materially alter the course of macrovascular disease, reduce health care costs, and improve the quality of life for people with NIDDM.


Reaction of phosphofructokinase 2/fructose 2,6-bisphosphatase with monoclonal antibodies. A proof of the bifunctionality of the enzyme.
Monoclonal antibodies were derived from mice immunized against homogeneous chicken liver phosphofructokinase 2/fructose 2,6-bisphosphatase. Of 112 clones, 30 were found to secrete antibodies that specifically reacted with the antigen in enzyme-linked immunoabsorbant assay (ELISA) while 17, which were ELISA-negative, produced antibodies that affected the enzymic activity of the antigen. Four clones were subcloned and used for an extensive investigation of the reaction of the corresponding antibodies with the supposedly bifunctional enzyme. A definite proof of the bifunctionality of the enzyme was obtained from the two following observations. First, the two activities were similarly retained by the four antibodies that had been coupled to Sepharose. Second, one of the antibodies inhibited both activities with the same efficiency. Furthermore, the antigen-antibody reaction led to the formation of aggregates with an apparent molecular mass of several megadaltons, showing that the two subunits of the antigen reacted with the same antibody and were therefore identical. The four monoclonal antibodies affected the activity of phosphofructokinase 2. This effect was seen as an up to 17-fold activation as well as an up to 85% inhibition. Only one of the four antibodies (antibody 10) had inhibitory effects on fructose 2,6-bisphosphatase, an effect which was in part explained by a decrease in the rate of formation of the intermediary phosphoenzyme. All the effects described above were obtained on both the chicken liver and the pigeon muscle enzymes but with lower doses of antibody in the case of the former enzyme. Antibody 10 was also shown to react with mouse liver phosphofructokinase 2/fructose 2,6-bisphosphatase, and with phosphofructokinase 2 from chicken brain, heart and testis and from frog skeletal muscle and liver. None of the four antibodies cross-reacted with phosphofructokinase 2 from Saccharomyces cerevisiae or from spinach leaves.


Resveratrol reverses the Warburg effect by targeting the pyruvate dehydrogenase complex in colon cancer cells.
Resveratrol (RES), a polyphenol found in natural foods, displays anti-oxidant, anti-inflammatory and anti-proliferative properties potentially beneficial in cancers, in particular in the prevention of tumor growth. However, the rapid metabolism of resveratrol strongly limits its bioavailability. The molecular mechanisms sustaining the potential biological activity of low doses of resveratrol has not been extensively studied and, thus, needs better characterization. Here, we show that resveratrol (10 µM, 48 hr) induces both a cell growth arrest and a metabolic reprogramming in colon cancer cells. Resveratrol modifies the lipidomic profile, increases oxidative capacities and decreases glycolysis, in association with a decreased pentose phosphate activity and an increased ATP production. Resveratrol targets the pyruvate dehydrogenase (PDH) complex, a key mitochondrial gatekeeper of energy metabolism, leading to an enhanced PDH activity. Calcium chelation, as well as the blockade of the mitochondrial calcium uniport, prevents the resveratrol-induced augmentation in oxidative capacities and the increased PDH activity suggesting that calcium might play a role in the metabolic shift. We further demonstrate that the inhibition of the CamKKB or the downstream AMPK pathway partly abolished the resveratrol-induced increase of glucose oxidation. This suggests that resveratrol might improve the oxidative capacities of cancer cells through the CamKKB/AMPK pathway.


Co-design of a Community-based Rehabilitation Program to Decrease Musculoskeletal Disabilities in a Mayan-Yucateco Municipality.
Chronic musculoskeletal (MSK) diseases are an important cause of disability in the Mayan community of Chankom in Yucatán, Mexico. To understand a community-based participatory research (CBPR) strategy implemented in Chankom to design a community-based rehabilitation (CBR) program for people living with MSK diseases. Qualitative descriptive thematic analysis from an ethnographic work conducted in Chankom, during the implementation of a CBPR strategy from 2014 to 2017. Four main themes describe the main processes that formed our CBPR strategy: 1) forming and maintaining an alliance between academic and community members, 2) prioritizing community needs, 3) integrating local and global knowledge and 4) shared-decision-making. This CBPR strategy allowed the design of a CBR program formed by six main interventions: 1) health services coordination, 2) personal support, 3) community venous blood sampling services, 4) community specialized services, 5) health promotion, and 6) health transportation services. Co-designing a CBR program for people living with chronic MSK diseases in Chankom was possible through an extensive community engagement work structured around four main processes, including the essential principles of CBPR. The designed CBR program includes culturally sensitive interventions aimed at improving the quality, availability, accessibility, and acceptability of health care services. Moreover, the program mainly addressed the "health" component of the World Health Organization-CBR matrix, suggesting a need for a new CBPR cycle after it is implemented and evaluated in the future.


There is (still) too much aluminium in infant formulas.
Infant formulas are sophisticated milk-based feeds for infants which are used as a substitute for breast milk. Historically they are known to be contaminated by aluminium and in the past this has raised health concerns for exposed infants. We have measured the aluminium content of a number of widely used infant formulas to determine if their contamination by aluminium and consequent issues of child health persists. Samples of ready-made milks and powders used to make milks were prepared by microwave digestion of acid/peroxide mixtures and their aluminium content determined by THGA. The concentration of aluminium in ready-made milks varied from ca 176 to 700 μg/L. The latter concentration was for a milk for preterm infants. The aluminium content of powders used to make milks varied from ca 2.4 to 4.3 μg/g. The latter content was for a soya-based formula and equated to a ready-to-drink milk concentration of 629 μg/L. Using the manufacturer's own guidelines of formula consumption the average daily ingestion of aluminium from infant formulas for a child of 6 months varied from ca 200 to 600 μg of aluminium. Generally ingestion was higher from powdered as compared to ready-made formulas. The aluminium content of a range of well known brands of infant formulas remains high and particularly so for a product designed for preterm infants and a soya-based product designed for infants with cow's milk intolerances and allergies. Recent research demonstrating the vulnerability of infants to early exposure to aluminium serves to highlight an urgent need to reduce the aluminium content of infant formulas to as low a level as is practically possible.


Induction of two transformation-sensitive membrane polypeptides in normal rat kidney cells by iron deprivation.
Rapid depletion of iron from the growth medium induces the synthesis of two membrane proteins with a subunit molecular weight of 160,000 (160K) and 130,000 (130K) in cultured normal rat kidney cells. When iron is maintained at normal levels in the growth medium of normal rat kidney cells, the synthesis of the 160K and 130K proteins is suppressed. We found that 160K and 130K are the underglycosylated forms of two membrane glycoproteins, 163K and 132K, respectively. We estimated the apparent turnover rate of the underglycosylated proteins and showed that it is slower than that of the fully glycosylated forms. Upon removal of iron from the growth medium pf simian virus 40-transformed normal rat kidney cells and Kirsten sarcoma virus-transformed normal rat kidney cells, 160K and 130K did not increase to levels comparable to those of normal rat kidney cells. Two different clones of simian virus 40-transformed normal rat kidney cells subjected to iron deprivation showed greatly reduced levels of 160K, and in one of these clones, 130K was absent. Kirsten sarcoma virus-transformed normal rat kidney cells also showed a defective response to iron deprivation manifested by reduced levels of both 160K and 130K. Additional studies indicate that these glycoproteins are membrane-associated procollagen molecules. The alteration in the coordinated induction of 160K and 130K in transformed cells suggests that these membrane-associated proteins may have an important role in transformation.


Methicillin-resistant Staphylococcus aureus isolates recovered from a New York City hospital: analysis by molecular fingerprinting techniques.
Fifty-five methicillin-resistant Staphylococcus aureus (MRSA) isolates collected at New York Hospital Medical Center of Queens in 1989 were analyzed by molecular fingerprinting techniques. Close to 70% of these isolates (38 of 55) shared a common pulsed-field gel electrophoretic pattern, carried the same mecA gene polymorph type II, were free of the transposon Tn554, and would not react with a mecI-specific gene probe. An additional five isolates shared all properties of the major MRSA clone except that they carried mecA gene polymorph type III. All these isolates had an extremely heterogeneous methicillin resistance phenotype that belonged to population analysis profile class 1 or 2. The rest of the 12 MRSA isolates showed a variety of chromosomal pulsed-field gel electrophoretic patterns that carried different mecA polymorphs and that also gave positive reactions with DNA probes for Tn554 and for the mecI gene. The molecular features of the majority MRSA clone suggest that it is an archaic MRSA isolate similar in features to early MRSA isolates recovered in the 1960s.