Immunization of mice with DNA-based Pfs25 elicits potent malaria transmission-blocking antibodies.
Immunological intervention, in addition to vector control and malaria chemotherapy, will be needed to stop the resurgence of malaria, a disease with a devastating impact on the health of 300 to 500 million people annually. We have pursued a vaccination strategy, based on DNA immunization in mice with genes encoding two antigens present on the sexual stages of Plasmodium falciparum, Pfs25 and Pfg27, to induce biologically important antibodies that can block development of the parasite in the Anopheles mosquito and thus transmission of the disease. DNA encoding Pfs25 when administered by the intramuscular route, either alone or with DNA encoding Pfg27, had the most potent transmission-blocking effects, resulting in up to a 97% decrease in oocyst numbers in mosquito midguts and a 75% decrease in rate of infection. Immunization with DNA encoding a Pfg27-Pfs25 fusion protein was less effective and DNA encoding Pfg27 elicited antibodies in sera that had only modest effects on the infectivity of the parasite. These results show for the first time that DNA vaccination can result in potent transmission-blocking antibodies in mice and suggest that the Pfs25 gene should be included as part of a multicomponent DNA vaccine.


Bax forms an oligomer via separate, yet interdependent, surfaces.
Interactions of Bcl-2 family proteins regulate permeability of the mitochondrial outer membrane and apoptosis. In particular, Bax forms an oligomer that permeabilizes the membrane. To map the interface of the Bax oligomer we used Triton X-100 as a membrane surrogate and performed site-specific photocross-linking. Bax-specific adducts were formed through photo-reactive probes at multiple sites that can be grouped into two surfaces. The first surface overlaps with the BH1-3 groove formed by Bcl-2 Homology motif 1, 2, and 3; the second surface is a rear pocket located on the opposite side of the protein from the BH1-3 groove. Further cross-linking experiments using Bax BH3 peptides and mutants demonstrated that the two surfaces interact with their counterparts in neighboring proteins to form two separated interfaces and that interaction at the BH1-3 groove primes the rear pocket for further interaction. Therefore, Bax oligomerization proceeds through a series of interactions that occur at separate, yet allosterically, coupled interfaces.


The effect of axial compression and distraction on cervical facet mechanics during anterior shear, flexion, axial rotation, and lateral bending motions.
The subaxial cervical facets are important load-bearing structures, yet little is known about their mechanical response during physiological or traumatic intervertebral motion. Facet loading likely increases when intervertebral motions are superimposed with axial compression forces, increasing the risk of facet fracture. The aim of this study was to measure the mechanical response of the facets when intervertebral axial compression or distraction is superimposed on constrained, non-destructive shear, bending and rotation motions. Twelve C6/C7 motion segments (70 ± 13 yr, nine male) were subjected to constrained quasi-static anterior shear (1 mm), axial rotation (4°), flexion (10°), and lateral bending (5°) motions. Each motion was superimposed with three axial conditions: (1) 50 N compression; (2) 300 N compression (simulating neck muscle contraction); and, (3) 2.5 mm distraction. Angular deflections, and principal and shear surface strains, of the bilateral C6 inferior facets were calculated from motion-capture data and rosette strain gauges, respectively. Linear mixed-effects models (α = 0.05) assessed the effect of axial condition. Minimum principal and maximum shear strains were largest in the compressed condition for all motions except for maximum principal strains during axial rotation. For right axial rotation, maximum principal strains were larger for the contralateral facets, and minimum principal strains were larger for the left facets, regardless of axial condition. Sagittal deflections were largest in the compressed conditions during anterior shear and lateral bending motions, when adjusted for facet side.


Effects of Rosiglitazone on the Outcome of Experimental Periapical Lesions in Mice.
The purpose of this study was to evaluate a protocol for systemic administration of rosiglitazone in mice in order to stimulate apoptosis of osteocytes in the jaws and to evaluate the effect of osteocyte apoptosis induced by rosiglitazone in the progression of periapical lesions in mice at 7, 21, and 42 days. C57BL/6 mice at 4-5 weeks of age were used. In phase 1, mice (n = 24) were treated with rosiglitazone (gavage, 10 mg/kg dose) or without (phosphate-buffered saline + 10% dimethyl sulfoxide) for 1, 2, or 3 weeks. We used the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and 4'-6-diamidino-2-phenylindole methods for quantification of apoptotic cells. In phase 2, mice (n = 30) received rosiglitazone for 2 weeks or just vehicle for 1 week (n = 30), and periapical lesions were induced for 7, 21, or 42 days. We performed the measurement of periapical lesions, tartrate-resistant acid phosphatase staining, dual-energy x-ray absorptiometry for the evaluation of bone mineral density (BMD) in long bone, and gene evaluation using real time quantitative polymerase chain reaction of osteocyte markers (Sost, Hyou1, and Dmp1) and receptor activator of nuclear factor kappa-B ligand (RANKL) (Tnfsf11). It was observed that systemic administration of rosiglitazone for 2 weeks showed apoptosis of osteocytes in a more expressive manner. In phase 2, in the groups that received rosiglitazone, a trend toward larger periapical lesions was observed (P > .05). Rosiglitazone also induced a greater number of osteoclasts and a greater expression of Sost and Hyou1 at 21 days of lesions. Moreover, there were no statistically significant differences in RANKL and Dmp1 expression or in the BMD of femurs. Rosiglitazone stimulated apoptosis of osteocytes, interfering in the progression of periapical lesions in mice.


Regenerative therapeutic potential of adipose stromal cells in early stage diabetic retinopathy.
Diabetic retinopathy (DR) is the leading cause of blindness in working-age adults. Early stage DR involves inflammation, vascular leakage, apoptosis of vascular cells and neurodegeneration. In this study, we hypothesized that cells derived from the stromal fraction of adipose tissue (ASC) could therapeutically rescue early stage DR features. Streptozotocin (STZ) induced diabetic athymic nude rats received single intravitreal injection of human ASC into one eye and saline into the other eye. Two months post onset of diabetes, administration of ASC significantly improved "b" wave amplitude (as measured by electroretinogram) within 1-3 weeks of injection compared to saline treated diabetic eyes. Subsequently, retinal histopathological evaluation revealed a significant decrease in vascular leakage and apoptotic cells around the retinal vessels in the diabetic eyes that received ASC compared to the eyes that received saline injection. In addition, molecular analyses have shown down-regulation in inflammatory gene expression in diabetic retina that received ASC compared to eyes that received saline. Interestingly, ASC were found to be localized near retinal vessels at higher densities than seen in age matched non-diabetic retina that received ASC. In vitro, ASC displayed sustained proliferation and decreased apoptosis under hyperglycemic stress. In addition, ASC in co-culture with retinal endothelial cells enhance endothelial survival and collaborate to form vascular networks. Taken together, our findings suggest that ASC are able to rescue the neural retina from hyperglycemia-induced degeneration, resulting in importantly improved visual function. Our pre-clinical studies support the translational development of adipose stem cell-based therapy for DR to address both retinal capillary and neurodegeneration.


Impacts of extension access and cooperative membership on technology adoption and household welfare.
This paper examines the impacts of access to extension services and cooperative membership on technology adoption, asset ownership and poverty using household-level data from rural Nigeria. Using different matching techniques and endogenous switching regression approach, we find that both extension access and cooperative membership have a positive and statistically significant effect on technology adoption and household welfare. Moreover, we find that both extension access and cooperative membership have heterogeneous impacts. In particular, we find evidence of a positive selection as the average treatment effects of extension access and cooperative membership are higher for farmers with the highest propensity to access extension and cooperative services. The impact of extension services on poverty reduction and of cooperatives on technology adoption is significantly stronger for smallholders with access to formal credit than for those without access. This implies that expanding rural financial markets can maximize the potential positive impacts of extension and cooperative services on farmers' productivity and welfare.


A protein-sialyl polymer complex involved in colominic acid biosynthesis. Effect of tunicamycin.
A protein-NeuAc complex involved in colominic acid biosynthesis has been identified in membrane preparations of Escherichia coli K-235. This compound had an Mr (estimated by SDS/polyacrylamide-gel electrophoresis and autoradiography) of about 100,000 and played the role of an 'initiator' or 'primer' (endogenous acceptor) in the synthesis of the whole polymer. Incubations of E. coli membranes with CMP-[14C]NeuAc (CMP-N-[14C]acetylneuraminic acid) pointed to the existence of a protein fraction (primer acceptor) that linked residues of sialic acid (N-acetylneuraminic acid, NeuAc) up to a maximal size, later releasing them as low-Mr sialyl polymers (LMrS, Mr less than 10,000). In the presence of colominic acid (final acceptor) the radioactivity linked to the protein quickly decreased, appearing stoichiometrically bound to the whole polysaccharide. When membrane preparations were previously digested with Streptomyces proteinase or de-activated by heating (80 degrees C, 10 min), no incorporation of labelled NeuAc into trichloroacetic acid-insoluble material was detected. These results suggested that colominic acid molecules are synthesized while they are bound to a proteinaceous acceptor that is subsequently excised in the presence of colominic acid, generating the native protein. The antibiotic tunicamycin inhibited the biosynthesis of colominic acid, affecting the synthesis of this protein-(NeuAc)n intermediate. All these results are described here for the first time.


Effects of 7-day amino acid infusion on renal growth, function, and renin-angiotensin system in fetal sheep.
These experiments examined whether renal growth and the fetal renin-angiotensin system could be stimulated by infusion of amino acids and whether chronic amino acid infusions restored glomerulotubular balance, which had been disrupted during 4-h infusions. Five fetal sheep aged 122 +/- 1 days gestation received an infusion of alanine, glycine, proline and serine in 0.15 M saline at 0.22 mmol/min for 7 days. Six control fetuses were given saline at the same rate (5 ml/h). Kidney wet weights after amino acid infusion were 28% larger than control fetuses (P < 0.05), and renal angiotensinogen mRNA levels were approximately 2.6-fold higher (P < 0.005). Circulating renin levels and renal renin mRNA levels were suppressed (P < 0.05), and renal renin protein levels tended to be lower. Arterial pressure was increased, and there was a marked, sustained natriuresis and diuresis. Glomerular filtration rate and filtered sodium were approximately two-fold higher throughout infusion (P < 0.05). Fractional proximal sodium reabsorption, suppressed at 4 h (from 73.4 +/- 6.5 to 53.7 +/- 10.2%), did not return to control levels (36.1 +/- 3.4% on day 7, P < 0.05). Distal sodium reabsorption was markedly increased (from 79 +/- 25 to 261 +/- 75 mumol/min by day 7, P < 0.005), but this was not sufficient to restore glomerulotubular balance. The resultant high rates of sodium excretion led to hyponatremia and polyhydramnios. In conclusion, long-term amino acid infusions increased renal angiotensinogen gene expression, kidney weight, and distal nephron sodium reabsorptive capacity but failed to restore proximal and total glomerulotubular balance.


Evaluation of Bond Strength of Cantilever Resin-retained Bridge Designs: An In Vitro Study.
The aim of the study was to compare the bond strength of three cantilever resin-bonded bridge (RBB) designs cemented with resin cements. Twenty-four extracted human mandibular canines with eight teeth per group were used in this study to evaluate the bond strength of cantilever resin-retained bridge designs of the mesh, perforated, and combination of mesh and perforated luted to the prepared lingual surface of canine teeth using resin cement. Debonding was done using Instron universal testing machine by applying load on the mid-buccal region of the pontic and the obtained values were evaluated. The data was analyzed statistically. The mean shear bond strengths of mesh, perforated, and combination of mesh and perforated are 0.88 ± 0.31 MPa, 0.81 ± 0.31 MPa, and 0.93 ± 0.32 MPa. However, there is no significant differences in the statistical analysis that were performed using a one-factor analysis of variant (ANOVA) test (p = 0.744). Within the limitation of this study for cantilever resin-retained bridge designs, the combination of mesh and perforated design showed greater mean shear bond values when compared with the mesh design and perforated designs. This shows equal in vitro performance to the gold standard designs (group A is mesh design and group B is perforated design). The new design which is the combination of mesh and perforated shows equal in vitro performance to the gold standard designs. Thus, their use in clinical situation can bring better result in concern to cantilever resin-retained prosthesis.


Major congenital heart defects are rarely diagnosed after newborns' hospital discharge with modern screening.
Our aim was to assess undiagnosed congenital heart defects (CHD) after newborns' hospital discharge in patients with a murmur or CHD suspicion, to find out the signs that predict CHDs and to estimate the costs of the examinations. We reviewed retrospective medical records of patients (n = 490) referred for the evaluation of CHD suspicion during 2017-2018. The median age of the patients was 2.5 (IQR 0.5-7.4) years. Sixty-three (13%) patients had an abnormal echocardiography. Neither ductal-dependent nor cyanotic CHDs were found. Cardiac interventions were performed for 14 out of 63 (22%) patients. Clinical signs indicating CHDs were murmur grade ≥3 (10/11 [91%] vs. 53/479 [11%], p < 0.001) and harsh murmur (15/44 [34%] vs. 48/446 [11%], p < 0.001). Abnormal electrocardiography did not indicate CHD (8/40 [20%] vs. 55/447 [12%], p = 0.165). The total cost of the examinations was 259 700€. The share of the cost of studies assessed as benign was 59%. Only a few CHDs were found after newborn hospital discharge among patients who received foetal and newborn screening and were examined due to CHD suspicion. The high number of benign murmurs in children leads to many referrals, resulting in unnecessary healthcare costs.


Distal renal tubular acidosis and its relationship with hearing loss in children: preliminary report.
In autosomal recessive distal renal tubular acidosis (DRTA), a substantial fraction of the patients have progressive bilateral sensorineural hearing loss. This coexistence is due to the mutations of a gene expressed both in the kidney and in the cochlea. The aim of this study was to assess the correlation between hearing loss and DRTA. In this study, 51 children diagnosed with renal tubular acidosis were evaluated. Diagnosis of DRTA was based on clinical manifestations and detection of normal anion gap metabolic acidosis, urine pH higher than 5.5, and positive urinary anion gap. Audiometry was performed in children with DRTA and sequencing of the ATP6V1B1 gene was done for those with sensorineural hearing loss. Twenty-seven patients (52.9%) had DRTA, of whom 51.9% were younger than 1 year old, 55.6% were boys, and 44.4% were girls. Eleven patients (40.7%) had bilateral sensorineural hearing loss, consisting of 5 of 15 boys (33.3%) and 6 of 12 girls (50.0%). There was no correlation between hearing loss and gender. Three patients with hearing loss had mutation in the ATP6V1B1 gene (11.1% of patients with DRTA and 27.3% of patients with DRTA and hearing loss). This study indicated that a significant percentage of the children with DRTA had sensorineural hearing loss and mutation in ATP6V1B1 gene. It is recommended to investigate hearing impairment in all children with DRTA.


Fluorescence identifies an alkaline cell in turtle urinary bladder.
Intracellular pH (pHi) of turtle bladder mucosal cells was studied by the trapped fluorescent indicator technique. Bladders efficiently accumulated and converted 4-methylumbelliferyl acetate to its pH-sensitive derivative 4-methylumbelliferone (4MU). Excited at the pH-indifferent wavelength 334 nm, bladders fluoresced a uniform blue. Using pH-sensitive 365-nm excitation, 10-20% of the mucosal cells fluoresced distinctly brighter, suggesting a more alkaline pHi. Using the 365/334 ratio to quantitate pHi, this difference averaged 0.1 pH units. Bright cells were more distinct after SITS or acetazolamide but disappeared after digitonin permeabilization, dinitrophenol, or treatment with propionate, DMO, and NH4Cl. Essentially the same population of bright cells was identified by carboxyfluorescein diacetate. The brighter cell corresponded exactly to a population of cells with distinctive acridine orange staining and bright costaining with the potential-sensing probes Di-O-C5, Di-S-C3, and 4-Di-5-Asp. Two extremes of bright cell shape were seen: an elongate cell, prevalent under conditions stimulating H+ secretion, and a more compact cell, when acidification was inhibited. These observations support the hypothesis that acidification represents H+ secretion via the luminal membrane and that a primary role of carbonic anhydrase in this process is to support the exit of base from the cell. The more alkaline cells appear to be the carbonic anhydrase-rich cells. These cells are chemically isolated from the surrounding granular cells and change their morphology in response to changes in acidification. These special properties indicate a unique role for the carbonic anhydrase cell in H+ secretion.


Mass casualty incidents: are NHS staff prepared? An audit of one NHS foundation trust.
Lack of knowledge of an NHS trust's major incident policies by clinical staff may result in poorly coordinated responses during a mass casualty incident (MCI). To audit knowledge of the major incident policy by clinical staff working in a central London major acute NHS trust designated to receive casualties on a 24-h basis during a MCI. A 12-question proforma was distributed to 307 nursing and medical staff in the hospital, designed to assess their knowledge of the major incident policy. Completed proformas were collected over a 2-month period between December 2006 and February 2007. A reply rate of 34% was obtained, with a reasonable representation from all disciplines ranging from nurses to consultants. Despite only 41% having read the policy in full, 70% knew the correct immediate action to take if informed of major incident activation. 76% knew the correct stand-down procedure. 56% knew the correct reporting point but less than 25% knew that an action card system was utilised. Nurses had significantly (p<0.01) more awareness of the policy than doctors. In view of the heightened terrorist threat in London, knowledge of major incident policy is essential. The high percentage of positive responses relating to immediate and stand-down actions reflects the rolling trust-wide MCI education programme and the organisational memory of the trust following several previous MCI in the capital. There is still scope for an improvement in awareness, however, particularly concerning knowledge of action cards, which are now displayed routinely throughout clinical areas and will be incorporated into induction packs.


Next-generation sequencing of microRNAs uncovers expression signatures in polarized macrophages.
microRNAs (miRNAs) are small noncoding RNAs that regulate gene expression at a posttranscriptional level and play a crucial role in the development of cells of the immune system. Macrophages are essential for generating inflammatory reactions upon tissue damage and encountering of invading pathogens, yet modulation of their immune responses is critical for maintaining tissue homeostasis. Macrophages can present different phenotypes, depending on the cytokine environment they encounter in the affected tissues. In this study, we have identified expression signatures of miRNAs that are differentially regulated during maturation of monocytes and polarization of macrophages by cytokines. We present a comprehensive characterization of miRNA expression in human monocytes and M1, M2a, and M2c polarized macrophages, using next-generation sequencing. Furthermore, we show that miRNA expression signatures are closely related to the various immune functions of polarized macrophages and therefore are involved in shaping the diverse phenotypes of these cells. The miRNAs identified here serve as markers for identification of inflammatory macrophages involved in the development of immune responses. Our findings contribute to understanding the role of miRNAs in determining the macrophage function in healthy and diseased tissues.


Saccharomyces cerevisiae Centromere RNA Is Negatively Regulated by Cbf1 and Its Unscheduled Synthesis Impacts CenH3 Binding.
Two common features of centromeres are their transcription into noncoding centromere RNAs (cen-RNAs) and their assembly into nucleosomes that contain a centromere-specific histone H3 (cenH3). Here, we show that Saccharomyces cerevisiae cen-RNA was present in low amounts in wild-type (WT) cells, and that its appearance was tightly cell cycle-regulated, appearing and disappearing in a narrow window in S phase after centromere replication. In cells lacking Cbf1, a centromere-binding protein, cen-RNA was 5-12 times more abundant throughout the cell cycle. In WT cells, cen-RNA appearance occurred at the same time as loss of Cbf1's centromere binding, arguing that the physical presence of Cbf1 inhibits cen-RNA production. Binding of the Pif1 DNA helicase, which happens in mid-late S phase, occurred at about the same time as Cbf1 loss from the centromere, suggesting that Pif1 may facilitate this loss by its known ability to displace proteins from DNA. Cen-RNAs were more abundant in rnh1Δ cells but only in mid-late S phase. However, fork pausing at centromeres was not elevated in rnh1Δ cells but rather was due to centromere-binding proteins, including Cbf1 Strains with increased cen-RNA lost centromere plasmids at elevated rates. In cbf1Δ cells, where both the levels and the cell cycle-regulated appearance of cen-RNA were disrupted, the timing and levels of cenH3 centromere binding were perturbed. Thus, cen-RNAs are highly regulated, and disruption of this regulation correlates with changes in centromere structure and function.


Role of the metal oxidation state in the SNS-Cr catalyst for ethylene trimerization: isolation of di- and trivalent cationic intermediates.
The reaction of the highly selective [CySCH2CH2N(H)CH2CH2SCy]CrCl3 catalyst precursor with alkyl aluminum activators was examined with the aim of isolating reactive intermediates. Reaction with Me3Al afforded a cationic trivalent chromium alkyl species {[CySCH2CH2N(H)CH2CH2SCy]CrMe(mu-Cl)}2{(AlMe3)2(m-Cl}2.(C7H8)2 (1a). Although it was not possible to obtain crystalline samples of sufficient quality from the reaction with MAO (the most preferred activator), the near-to-identical EPR spectra indicated a very close structural similarity with 1a. Ethylene oligomerization tests clearly revealed that 1 and other cationic trivalent dimeric complexes {[CySCH2CH2N(H)CH2CH2SCy] CrCl(mu-Cl)}2{AlCl4}2.(C7H8)1.5 (2), monomeric [(CySCH2CH2N(H)CH2CH2SCy)CrCl2 (THF)][AlCl4] (3), and {[CySCH2CH2N(H)CH2CH2SCy]Cr(eta2-AlCl4)}{Al2Cl7} (4) adducts display the same catalyst selectivity as the [CySCH2CH2N(H)CH2CH2SCy]CrCl3 complex and, therefore, are probably all precursors to the same catalytically active species. 2, 3, and 4 were obtained upon treatment of [CySCH2CH2N(H)CH2CH2SCy] CrCl3 with different stoichiometric ratios of AlCl3.. When i-BAO activator was used, reduction of the metal center occurred readily, affording {([CySCH2CH2N(H)CH2CH2S Cy]Cr)(mu-Cl)]2}{(i-Bu)2AlCl2}2 (5). 5 is also a selective catalyst, thus indicating that trivalent species are most probably precursors to a divalent catalytically active complex. Reaction of CrCl2(THF)2 with the ligand afforded the labile divalent adduct [CySCH2CH2N(H)CH2CH2SCy]CrCl2(THF) (6), also catalytically active and selective. Instead, deprotonation of the ligand with n-BuLi followed by reaction with CrCl2(THF)2 gave the dinuclear complex [(mu-CySCH2CH2NCH2CH2SCy)CrCl]2 (7), which did not produce oligomers.


Isoalantolactone Enhances the Radiosensitivity of UMSCC-10A Cells via Specific Inhibition of Erk1/2 Phosphorylation.
Although radiotherapy is one of the mainstream approaches for the treatment of head and neck squamous cell carcinoma (HNSCC), this cancer is always associated with resistance to radiation. In this study, the mechanism of action of isoalantolactone as well as its radiosensitizing effect was investigated in UMSCC-10A cells. The radiosensitization of UMSCC-10A cells treated with isoalantolactone was analyzed by colony formation assay. The radiosensitization effects of isoalantolactone on cell proliferation, cell cycle and apoptosis regulation were examined by BrdU incorporation assay, DNA content assay and flow cytometry, respectively. Western blotting was performed to determine the effects of isoalantolactone combined with radiation on the protein expression of Mek, extracellular signal-regulated kinase (Erk1/2) as well as phosphorylated Mek and Erk1/2. Erk1/2 knockdown by siRNA was used to confirm that isoalantolactone specifically inhibited the activation of Erk1/2 signaling pathway in UMSCC-10A cells. Isoalantolactone enhanced the radiosensitivity of UMSCC-10A cells; the sensitivity enhanced ratios (SERs) were 1.44 and 1.63, respectively, for 2.5 and 5 μM. Moreover, isoalantolactone enhanced radiation-induced cell proliferation and apoptosis and cell cycle arrested at G2/M phase. Furthermore, no marked changes were observed in the expression of total Erk1/2 and Mek protein after radiation treatment. However, isoalantolactone was significantly reduced radiation-induced the phosphorylation of Erk1/2, whereas it altered the phosphorylation of Mek to a lesser extent. In addition, the radiosensitivity of UMSCC-10A cells with Erk1/2 knockdown was increased. Isoalantolactone cannot further prevent the proliferation of UMSCC-10A cells with Erk1/2 knockdown which other mechanism regulated cell proliferation. Our results suggested that isoalantolactone enhanced radiation-induced apoptosis, cell cycle arrested and reduced the cell proliferation of UMSCC-10A cells via specifically inhibited the phosphorylation of Erk1/2. Thus a low concentration of isoalantolactone may be used to overcome the resistance of UMSCC-10A cells to radiation and may be a promising radiosensitizer in cancer therapy.


Can prognostic indicators be identified in a fetus with an encephalocele?
Encephaloceles, like other congenital malformations of the brain diagnosable in utero, can be either complicated (there being an associated chromosomal abnormality, abnormalities in the remainder of the central nervous system (CNS) and/or other organs), or isolated (no abnormalities in the chromosomes, the remainder of the CNS or other organs). Complicated cases invariably have a poor prognosis but amongst those with isolated lesions the outcome is variable with some affected children having poor mental and physical development but others who are only mildly or moderately disabled. To be able to make an informed decision about how to manage their pregnancy parents need to know what the prognosis is likely to be for their fetus with an encephalocele. To see if the necessary information could be reliably gathered by prenatal assessment of affected fetuses, a review was carried out of the medical records and ultrasound scans of 31 fetuses with encephaloceles referred to the Fetal Management Unit at St. Mary's Hospital in Manchester between January 1991 and December 1997. Eighteen of the cohort were classified as having a complicated encephalocele. Thirteen of the pregnancies were terminated; there were three intrauterine deaths, and one neonatal death. There is only one surviving child who is severely disabled. Thirteen fetuses were classified as having isolated encephaloceles, six had a mass of neural tissue in the encephalocele sac and were terminated, one died in utero and six had a cystic lesion or only a nubbin and have survived with few or no abnormalities. This study has shown that it is possible to identify fetuses with an encephalocele with a favourable outcome.


Feasibility of negative pressure wound therapy during intercontinental aeromedical evacuation of combat casualties.
The objective of this study was to assess the feasibility of utilizing negative pressure wound therapy (NPWT) for the treatment of wartime soft-tissue wounds during intercontinental aeromedical evacuation. Attempts to use NPWT during early phases of overseas contingency operations resulted in occasional vacuum system failures and potentially contributed to wound complications. These anecdotal episodes led to a perception that NPWT during aeromedical evacuation carried a high risk of wound complications and limited its use. As a result, NPWT was not frequently applied in the management of soft-tissue wounds before US casualty arrival in the continental United States (CONUS) for wounds sustained in the combat theaters. Concurrently, early NPWT on the traumatic wounds of host nation casualties not requiring aeromedical evacuation seemed to provide many benefits typically associated with the therapy such as decreased infection rates, earlier wound closure, and improved pain management. On a daily basis, study investigators reviewed the trauma in-patient census at Landstuhl Regional Medical Center, Germany, to identify patient candidates with soft-tissue extremity or torso wounds that required packing. Patient demographics, injuries, and previous wound treatments were recorded. Surgeons inspected wounds in the operating room and applied a NPWT dressing if deemed appropriate. NPWT was continued throughout the remainder of the patient's hospitalization and also during aeromedical evacuation to CONUS. A study investigator escorted the patient during aeromedical evacuation to educate the flight crews, to record the impact on crew workload, and to troubleshoot the system if necessary. Thirty enrolled patients with 41 separate wounds flew from Germany to CONUS with a portable NPWT system (VAC Freedom System; Kinetic Concepts Incorporated, San Antonio, TX). All 30 patients arrived at the destination facilities with intact and functional systems. No significant in-flight complications were identified, impact on flight crew workload was negligible, and subjective feedback from both flight crews and patients was uniformly positive. For 29 patients, the NPWT dressing was replaced (frequently with serial exchanges) during initial surgical treatment in CONUS; the 30th patient underwent delayed primary closure of his right forearm fasciotomy. Receiving care teams reported no complications attributable to NPWT during aeromedical evacuation. NPWT is feasible during intercontinental aeromedical evacuation of combat casualties without an increase in wound complications or a significant impact on air crew workload. Further studies are indicated to evaluate the efficacy of NPWT in combat wounds compared with other wound care techniques.


Comparative labellar micromorphology of Zygopetalinae (Orchidaceae).
Molecular evidence indicates that the Neotropical sub-tribe Zygopetalinae is sister to Maxillariinae. Most members of the latter sub-tribe have deceit pollination strategies, but some species produce rewards such as nectar, pseudopollen, resin and wax, and are pollinated by a range of pollinators that include stingless bees (Meliponini), wasps and hummingbirds. By contrast, relatively little is known about the pollination of Zygopetalinae species. However, some are pollinated by fragrance-gathering, male euglossine bees or employ nectar deceit strategies. The aim of this study is to describe the labellar micromorphology of Zygopetalinae and to compare it with that of Maxillariinae sensu lato (s.l.) as part of an ongoing project to record the range of labellar characters found within the tribe Maxillarieae, and to assess whether these characters represent synapomorphies or homoplasies resulting from similar pollination pressures. The labella of 31 species of Zygopetalinae, including Cryptarrhena R. Br. and representatives of the Zygopetalum, Huntleya and Warrea clades, were examined using light microscopy and scanning electron microscopy, and the range of labellar characters was recorded. These characters were subsequently compared with those of Maxillariinae s.l. which formed the subject of our previous investigations. The labellar micromorphology of Zygopetalinae is less diverse than that of Maxillariinae and does not reflect the currently accepted phylogeny of the former sub-tribe based on molecular studies. Instead, the relative uniformity in labellar micromorphology of Zygopetalinae is probably due to homoplasies resulting from similar pollinator pressures. Labellar trichomes are relatively uncommon in Zygopetalinae, but occur in certain members of both the Zygopetalum and Huntleya clades. Trichomes are unbranched, uniseriate and multicellular with rounded apices, or unbranched and unicellular, with tapering, pointed and flexuose apices. Hitherto, unicellular trichomes of this kind have been observed only for euglossophilous orchid taxa, and the adoption of a relatively limited range of pollination strategies by Zygopetalinae may have resulted in reduced investment in micromorphological labellar characters.


Shipping fever pasteurellosis and salmonellosis prophylaxis.
A water-in-oil emulsion adjuvanted combination shipping fever bacterin containing Pasteurella multocida, P hemolytica, Salmonella typhimurium and certain other species was effective in field use as a prophylactic agent against shipping fever pasteurellosis and salmonellosis. In trials using ranch calves it produced a greater antibody response than a comparable aluminum hydroxide adjuvanted bacterin. In critical 30-hour postvaccination challenge mouse tests it was superior to the conventional type bacterin and induced rapid and remarkable protection.


[Comparative analysis of molecular genetic studies using DNA samples isolated from blood treated with EDTA or heparin].
A molecular genetic investigation of five genes: CFTR, GSTM1, GCTT1, MTHFR, and TP53 using DNA samples isolated from blood treated with either EDTA or stabilized heparin was conducted. Twenty DNA samples obtained from 10 persons. The results prove that heparin used as an anticoagulant instead of EDTA had no inhibitory effect on PCR and PCR/RFLP results, at least when DNA was isolated with a DNA-sorb-B kit. The results suggest that heparin-treated blood samples can be also used to study other genes.


Emerging trends in the inhibition of bacterial molecular communication: An overview.
Quorum sensing (QS) is a molecular cell-cell communication utilized by several bacteria and some fungi. It involves cell density dependent gene expression that includes extra polymeric substance production, sporulation, antibiotic production, motility, competence, symbiosis and conjugation. These expressions were carried out by different signaling molecules like acyl homo-serine lactone (AHL) and auto-inducing peptides (AIPs) which was effluxed by gram negative and gram positive bacteria. Pathogenic bacteria and biofilms often exhibit high resistance to antibiotics, attributed to the presence of antibiotic efflux pumps, reduced membrane permeability, and enzymes that deactivate quorum sensing (QS) inhibitors. To counteract virulence and multi-drug resistance (MDR), novel strategies such as employing quorum sensing (QS) inhibitors and quorum quenchers are employed. It targets signaling molecules with synthesis and prevents the signal from binding to receptors. In this present review, the mechanisms of QS along with inhibitors from different sources are described. These strategies potentially interfere with QS and it can be applied in different fields, mainly in hospitals and marine environments where the pathogenic infections and biofilm formation are highly involved.


Amniotic fluid heat shock protein 70 concentration in histologic chorioamnionitis, term and preterm parturition.
Heat shock protein (HSP) 70, a conserved member of the stress protein family, is produced in almost all cell types in response to a wide range of stressful stimuli, and its production has a survival value. Evidence suggests that extracellular HSP70 is involved in the activation of the innate and adaptive immune response. Furthermore, increased mRNA expression of HSP70 has been observed in human fetal membranes following endotoxin stimulation. This study was conducted to determine the changes in amniotic fluid HSP70 concentrations during pregnancy, term and preterm parturition, intra-amniotic infection (IAI), and histologic chorioamnionitis. A cross-sectional study was conducted in 376 pregnant women in the following groups: (1) women with a normal pregnancy who were classified into the following categories: (a) women in the mid-trimester (14-18 weeks) who underwent amniocentesis for genetic indications and delivered normal infants at term (n=72); (b) women at term not in labor (n = 23); and (c) those at term in labor (n = 48). (2) Women with spontaneous preterm labor and intact membranes who were subdivided into the following categories: (a) preterm labor who delivered at term without IAI (n = 42); (b) preterm labor who delivered preterm without IAI (n = 57); and (c) preterm labor and delivery with IAI (n = 30). (3) Women with preterm prelabor rupture of membranes (PROM) with (n = 50) and without (n = 54) IAI. Among patients with preterm labor with intact membranes and preterm PROM who delivered within 72 hours of amniocentesis, placenta, umbilical cord, and chorioamniotic membranes were collected and assessed for the presence or absence of acute inflammatory lesions in the extraplacental membranes (histologic chorioamnionitis) and/or umbilical cords (funisitis). HSP70 concentrations in amniotic fluid were determined using a sensitive and specific immunoassay. Non-parametric statistics were used for analysis. A p value of <0.05 was considered statistically significant. Immunoreactive HSP70 was detected in 88% (332/376) of amniotic fluid samples. The median amniotic fluid HSP70 concentration was significantly higher in women at term without labor than in those in the mid-trimester (term no labor: median 34.9 ng/mL, range 0-78.1 ng/mL vs. mid-trimester; median 6.6 ng/mL, range 0-20.8 ng/mL; p<0.001). Among patients with spontaneous preterm labor and preterm PROM, those with IAI had a significantly higher median amniotic fluid HSP70 concentration than those without IAI (preterm labor with IAI: median 82.9 ng/mL, range 0-500 ng/mL vs. preterm labor without IAI: median 41.7 ng/mL, range 0-244 ng/mL; p = 0.001; preterm PROM with IAI: median 86.5 ng/mL, range 0-428 ng/mL vs. preterm PROM without IAI: median 55.9 ng/mL, range 14.9-299.9 ng/mL; p = 0.007). There was no significant difference in the median amniotic fluid HSP70 concentration between patients with preterm labor who delivered preterm without IAI and those who delivered at term (p = 0.6). However, among patients with preterm labor without IAI, there was an inverse relationship between amniotic fluid concentration of HSP70 and the amniocentesis-to-spontaneous delivery interval (Spearman's Rho = -0.26; p = 0.02). Patients with histologic chorioamnionitis/funisitis had a significantly higher median amniotic fluid HSP70 concentration than those without inflammation (inflammation: median 108.7 ng/mL, range 0-500 ng/mL vs. without inflammation: median 67.9 ng/mL, range 7.1-299.9 ng/mL; p = 0.02). Women at term in labor had a median amniotic fluid concentration of HSP70 significantly higher than those not in labor (term in labor: median 60.7 ng/mL, range 0-359.9 ng/mL vs. term not in labor: median 34.9 ng/mL, range 0-78.1 ng/mL; p = 0.02). Intra-amniotic infection, histologic chorioamnionitis, and term parturition are associated with elevated amniotic fluid HSP70 concentrations. HSP70 plays a role in the host defense mechanism by activating the innate arm of the immune response in women with intrauterine infection. The mechanisms of preterm and term parturition in humans may involve extracellular HSP70.


[Systematic approach to detection of anti-idiotypic antibodies to antimycobacterial idiotype in man and animals].
ELISA in the sandwich modification was developed to detect anti-idiotypic antibodies to the antituberculosis idiotype of monoclonal antibodies (MAb) S4C1G4 (anti 15-18 kDa H37Rv). Sera immunoglobulins were determined by Staphylococcus protein A capture. F[ab+]2, the MAb fragments S4C1G4 (IgG2a, kappa) and 1.3.3.B5 (IgG2a, kappa), were used as a negative control, which were not recognized mycobacterial antigens and human immunoglobulins. The anti-idiotypic antibody titers were determined in the murine hyperimmune sera during the period of immunization with the MSAb SaC1G4. The method detected anti-idiotypic antibodies in the sera of infected mice and in patients with pulmonary tuberculosis. It is suggested that this method is applicable while using diagnostic tools.


Genetic-Quantitative Study of the First-Service Pregnancy Probability of Murrah Heifers.
Because of the importance of reproduction in stock breeding systems, it is necessary to find selection criteria that increase reproductive efficiency. The aim of this study was to estimate genetic parameters for the probability of conception on first service (PROB) in Murrah heifers, and its association with other traits of economic interest [age at first calving (AFC), service period, calving interval and milk yield at 270 days], with the purpose of evaluating their use as selection criteria. Reproductive information and first lactation records of 1200 Murrah heifers were used to perform two-trait analyses between PROB and the other characteristics. Bayesian inference was used to estimate the variance components, considering PROB as threshold and the other as linear factors. The results demonstrate that this trait has heritability of 0.15, indicating the possibility of a genetic gain by using it for selection. With respect to the genetic correlation estimates, the only high-magnitude association was with AFC (-0.899), which is the current criterion indicating sexual precocity of females. In the light of the parameters estimated, the first-service pregnancy rate is an alternative for indication of sexual precocity, although presenting a smaller genetic gain than the current standard AFC. Nevertheless, additional research should be conducted regarding this trait to assess the economic importance of its use in dairy buffalo production systems.


Actinomadura madurae causing mycetomas in Madras.
Two hundred and ten histologically diagnosed cases of mycetoma filed in the Departments of Pathology of four Medical Colleges in Tamil Nadu, India, between 1964-1987 were subjected to detailed histopathological study; Actinomadura madurae, was found in 73 of them. In nine cases of mycetoma pedis, the organism A. madurae has been isolated in pure culture. In one case, the lesions were present in both the feet. The characteristic cream-coloured granules were seen in all. The lesions responded well to a combination of Streptomycin and dapsone.


Contactless respiratory and heart rate monitoring: validation of an innovative tool.
To assess the accuracy of the EverOn™ piezoelectric sensor based contactless heart rate and respiration rate monitoring system. Measurements of the EverOn™ and reference devices were performed in a sleep lab and an intensive care unit (ICU) setting. One minute measurements by both the reference device and the EverOn™ were averaged and compared. Accuracy was defined in accordance with industry criteria. Respiration rate (RR) accuracy in the 41 children and 16 adults evaluated in the sleep lab was 93.1% and 90.6% respectively, and heart rate (HR) accuracy was 94.4% and 91.5% respectively. For the 42 ICU patients RR accuracy was 82.0% and 75% (versus end-tidal CO(2) and manual respectively), while accuracy of HR was 94.0%. The EverOn™ was found to be superior to the impedance technique in measuring RR. The system described was found to be accurate in accordance with regulatory and industry criteria.


Dynamically Reconfigurable Metadevice Employing Nanostructured Phase-Change Materials.
Mastering dynamic free-space spectral control and modulation in the near-infrared (NIR) and optical regimes remains a challenging task that is hindered by the available functional materials at high frequencies. In this work, we have realized an efficient metadevice capable of spectral control by minimizing the thermal mass of a vanadium dioxide phase-change material (PCM) and placing the PCM at the feed gap of a bow-tie field antenna. The device has an experimentally measured tuning range of up to 360 nm in the NIR and a modulation depth of 33% at the resonant wavelength. The metadevice is configured for integrated and local heating, leading to faster switching and more precise spatial control compared with devices based on phase-change thin films. We envisage that the combined advantages of this device will open new opportunities for signal processing, memory, security, and holography at optical frequencies.


Comparison of CID versus ETD based MS/MS fragmentation for the analysis of protein ubiquitination.
Ubiquitination has emerged as one of the major post-translational modifications that decide on protein fate, targeting, and regulation of protein function. Whereas the ubiquitination of proteins can be monitored with classic biochemical methods, the mapping of modified side chains proves to be challenging. More recently, mass spectrometry has been applied to identify ubiquitinated proteins and also their sites of modification. Typically, liquid chromatography tandem mass spectrometry (LC-MS/MS) based approaches, including collision-induced fragmentation (CID), have been successfully used in the past. However, a potential difficulty arises from the unstable nature of this modification, and also that the isopeptide bond linkage between C-terminal glycine and the N(epsilon) lysyl side chain is susceptible to fragmentation under these conditions. Here we investigate the utility of electron-transfer dissociation (ETD)-based fragmentation to detect ubiquitination sites in proteins. Our results indicate that ETD can provide alternative fragmentation patterns that allow detection of gly-gly-modified lysyl side chains, in particular z+1 fragment ions derived from triply charged precursor ions. We subsequently applied ETD fragmentation-based analysis and detected novel ubiquitination sites on DNA polymerase B1 that were not easily observed using CID. We conclude that ETD can provide significant alternative fragmentation information that complements CID-derived data to improve the coverage when mapping ubiquitination sites in proteins.


Rabbit: frequency of suckling in the pup.
Rabbit does were given free access to their young, access once a day, and access twice a day. In all three groups the young were nursed only once every 24 hours. Growth curves for the pups from day 2 of age to day 30 were identical for all three groups. The restriction of suckling to once a day appears to depend on the mother and not the pups.


DPAGT1-CDG: report of a patient with fetal hypokinesia phenotype.
Congenital disorders of glycosylation (CDG) are due to either defects in the synthesis of the glycan moiety of glycoproteins or glycolipids and in the attachment of the glycans to proteins and lipids. Some 50 CDG have been identified. They represent a challenge for clinicians because most are multisystem diseases with a heterogeneous spectrum of clinical manifestations with involvement of any organ and system. We report on a patient with a mutation in the glycosyltransferase encoded by the DPAGT1 gene, an infrequent CDG. He showed severe fetal hypokinesia phenotype with decreased fetal movements and polyhydramnios. At birth he showed decreased facial expression, without nasolabial folds, soft long ears, U-shaped vermilion of the upper lip, thick skin, hypertrichosis, camptodactyly, moderate multiple contractures, hypotonia and severe hypokinesia, no spontaneous movements, and very limited movements with stimuli; he died at 1½ months. Isoelectrofocusing of serum transferrin showed a type 1 pattern with increased asialo- and disialotransferrin. The study of the DPAGT1 gene showed he was a compound heterozygote for two novel point missense mutations [c.901C>T]+[c.1094T>G]. This phenotype expands the clinical features of the few DPATG1-CDG patients reported.


Chinese preschoolers' resource allocation in the face of existing inequality under collaborative and noncollaborative contexts.
The current study investigated the stage at which Chinese preschoolers started considering recipients' material welfare and minimizing existing inequalities under both noncollaborative and collaborative contexts. Also, it analyzed how they behaved when recipients' material welfare was in conflict with merit or equality rule. Experiment 1 found that 4-year-old children allocated resources to the poor rather than the rich character only under the noncollaborative context but not under the collaborative context, in which they had observed these 2 characters working together to finish a task. By contrast, 5-year-old children started doing so under both collaborative and noncollaborative contexts. When there was conflict between recipients' material welfare and merit in Experiment 2, 6-year-old children did not show any preference for either material welfare or merit, and they made equal or almost equal distributions. Without collaboration information, Experiment 3 further revealed that the children aged from 4 to 6 y preferred equal distribution to resource allocation considering recipients' material welfare. These findings have been discussed in terms of contextual information and cultural influences. (PsycINFO Database Record


Characterization of human pluripotent stem cell differentiation by single-cell dual-omics analyses.
In vivo differentiation of human pluripotent stem cells (hPSCs) has unique advantages, such as multilineage differentiation, angiogenesis, and close cell-cell interactions. To systematically investigate multilineage differentiation mechanisms of hPSCs, we constructed the in vivo hPSC differentiation landscape containing 239,670 cells using teratoma models. We identified 43 cell types, inferred 18 cell differentiation trajectories, and characterized common and specific gene regulation patterns during hPSC differentiation at both transcriptional and epigenetic levels. Additionally, we developed the developmental single-cell Basic Local Alignment Search Tool (dscBLAST), an R-based cell identification tool, to simplify the identification processes of developmental cells. Using dscBLAST, we aligned cells in multiple differentiation models to normally developing cells to further understand their differentiation states. Overall, our study offers new insights into stem cell differentiation and human embryonic development; dscBLAST shows favorable cell identification performance, providing a powerful identification tool for developmental cells.


Bacteriophage P22-mediated specialized transduction in Salmonella typhimurium: high frequency of aberrant prophage excision.
The temperate bacteriophage P22 mediates both generalized and specialized transduction in Salmonella typhimurium. Specialized transduction by phage P22 is different from, and less restricted than, the well characterized specialized transduction by phage lambda, due to differences in the phage DNA packaging mechanisms. Based on the properties of the DNA packaging mechanism of phage P22 a model for the generation of various types of specialized transducing particles is presented that suggests generation of substantial numbers of specialized transducing genomes which are heterogeneous but only some of which have terminally redundant ends. The primary attachment site, ataA, for phage P22 in S. typhimurium is located between the genes proA,B and supQ newD. (The newD gene is a substitute gene for the leuD gene, restoring leucine prototrophy of leuD mutant strains.) The proA,B and supQ newD genes are very closely linked and thus cotransducible by generalized transducing particles. Specialized transducing particles can carry either proA,B or supQ newD but not both simultaneously, and thus cannot give rise to cotransduction of the proA,B and supQ newD genes. This difference is used to calculate the frequency of generalized and specialized transducing particles from the observed cotransduction frequency in phage lysates. By this method, very high frequencies of supQ newD (10(-2)/PFU)- and proA,B (10(-3)/PFU)-specialized transducing particles were detected in lysates produced by induction of lysogenic strains. These transducing particles most of which would have been produced by independent aberrant excision events (which include in situ packaging), were of various types.


Germ-cell tumors of the mediastinum.
Germ-cell tumors occurring primarily in the mediastinum have been well recognized in the literature for many years. Unfortunately, most of what has been written on these tumors has been in the form of case reports and small series of cases, thus precluding a better understanding of their clinical and demographic features, as well as of their biologic behavior in this particular location. In addition, mediastinal germ-cell tumors not only have never had a standardized nomenclature, but they also have lacked the benefit of a clinical staging scheme for assessment of prognosis. Another important aspect of these tumors that has required attention is the frequency with which teratomatous lesions are associated with malignant components in the mediastinum. Comprehensive studies of mediastinal germ-cell tumors to determine whether these lesions recapitulate the distribution, histopathologic features, and biologic behavior of their counterparts in other areas, such as the gonads, have been lacking for many years. In this article, we review our experience with a large series of cases of primary mediastinal germ cell tumors and examine recent concepts and advances in our understanding of these unusual and diagnostically challenging neoplasms.


Identification of functional microRNAs released through asymmetrical processing of HIV-1 TAR element.
The interaction between human immunodeficiency virus type 1 (HIV-1) and RNA silencing pathways is complex and multifaceted. Essential for efficient viral transcription and supporting Tat-mediated transactivation of viral gene expression, the trans-activation responsive (TAR) element is a structured RNA located at the 5' end of all transcripts derived from HIV-1. Here, we report that this element is a source of microRNAs (miRNAs) in cultured HIV-1-infected cell lines and in HIV-1-infected human CD4+ T lymphocytes. Using primer extension and ribonuclease (RNase) protection assays, we delineated both strands of the TAR miRNA duplex deriving from a model HIV-1 transcript, namely miR-TAR-5p and miR-TAR-3p. In vitro RNase assays indicate that the lack of a free 3' extremity at the base of TAR may contribute to its low processing reactivity in vivo. Both miR-TAR-5p and miR-TAR-3p down-regulated TAR miRNA sensor activity in a process that required an integral miRNA-guided RNA silencing machinery. miR-TAR-3p exerted superior gene downregulatory effects, probably due to its preferential release from HIV-1 TAR RNA by the RNase III Dicer. Our study suggests that the TAR element of HIV-1 transcripts releases functionally competent miRNAs upon asymmetrical processing by Dicer, thereby providing novel insights into viral miRNA biogenesis.


Flocculation of polymer stabilized nanocrystal suspensions to produce redispersible powders.
Aqueous suspensions of crystalline naproxen nanoparticles, formed by antisolvent precipitation, were flocculated with sodium sulfate, filtered, and dried to form redispersible powders for oral delivery. The particles were stabilized with polyvinylpyrrolidone (PVP K-15) and/or poly(ethylene oxide-b-propylene oxide-b-ethylene oxide) (poloxamer 407). The yield of the drug in the powder was typically 92-99%, and the drug loading was reproducible to within 1-2%. The filtration process increased the drug loading by up to 61% relative to the initial value, as unbound surfactant was removed with the filtrate. Upon redispersion of the dried powder, the average particle size measured by light scattering was comparable to the original value in the aqueous suspension prior to flocculation, and consistent with primary particle sizes observed by scanning electron microscopy (SEM). For 300-nm particles, up to 95% of the drug dissolved in 2 min. The dissolution rate was correlated linearly with the specific surface area calculated from the average particle diameter after redispersion. The redispersion of dried powders was examined as a function of the salt concentration used for flocculation and the surfactant composition and concentration. Flocculation followed by filtration and drying is an efficient and highly reproducible process for the rapid recovery of drug nanoparticles to produce wettable powders with high drug loading and rapid dissolution.


[Study of the mesenteric arteries with multislice spiral CT angiography].
The aim of our study was to optimize an high-resolution CT angiography protocol for the evaluation of mesenteric arteries using a multislice spiral CT equipment. A volumetric dataset was acquired during dynamic i.v. injection of non-ionic iodine contrast medium; images were subsequently reconstructed by means of a dedicated off-line workstation, equipped with volume-rendering software. Excellent depiction of the anatomy of mesenteric arteries and collateral vessels was obtained in all the cases. In conclusion, new multislice spiral CT scanners allow an optimal noninvasive evaluation of mesenteric arteries, possible up to now only with catheter angiography.


Synthesis of hyaluronate in differentiated teratocarcinoma cells. Characterization of the synthase.
Differentiation of teratocarcinoma cells led to induction of hyaluronate synthesis. The synthase was recovered in the membrane fraction of cell lysates. Hyaluronate was synthesized at the membranes and was then released as a soluble product. The synthase could be stimulated by a variety of phosphate esters which prevented the degradation of the substrates UDP-GlcNAc and UDP-GlcA and the release of the growing hyaluronic acid chain from the membrane. Hyaluronidases or oligosaccharides derived from hyaluronate did not affect the synthesis. The chains grew at a rate of 60 repeating units/min. Continuous new chain initiation occurred during prolonged synthesis. Digestion of pulse-chase-labelled hyaluronate with beta-N-acetylglucosaminidase and beta-glucuronidase showed that the chains grew at the reducing end.


Alteration of a single amino acid changes the substrate specificity of dihydroflavonol 4-reductase.
Many plant species exhibit a reduced range of flower colors due to the lack of an essential gene or to the substrate specificity of a biosynthetic enzyme. Petunia does not produce orange flowers because dihydroflavonol 4-reductase (DFR) from this species, an enzyme involved in anthocyanin biosynthesis, inefficiently reduces dihydrokaempferol, the precursor to orange pelargonidin-type anthocyanins. The substrate specificity of DFR, however, has not been investigated at the molecular level. By analyzing chimeric DFRs of Petunia and Gerbera, we identified a region that determines the substrate specificity of DFR. Furthermore, by changing a single amino acid in this presumed substrate-binding region, we developed a DFR enzyme that preferentially reduces dihydrokaempferol. Our results imply that the substrate specificity of DFR can be altered by minor changes in DFR.


The effects of Mg2+ on certain steps in the mechanisms of the dehydrogenase and esterase reactions catalysed by sheep liver aldehyde dehydrogenase. Support for the view that dehydrogenase and esterase activities occur at the same site on the enzyme.
Stopped-flow experiments in spectrophotometric and fluorescence modes reveal different aspects of the aldehyde dehydrogenase mechanism. Spectrophotometric experiments show a rapid burst of NADH production whose course is not affected by Mg2+. The slower burst seen in the fluorescence mode is markedly accelerated by Mg2+. It is argued that the fluorescence burst accompanies acyl-enzyme hydrolysis and, therefore, that Mg2+ increases the rate of this process. Experiments on the hydrolysis of p-nitrophenyl propionate indicate that acyl-enzyme hydrolysis is indeed accelerated by Mg2+ and a combination of Mg2+ and NADH. Vmax. values for p-nitrophenyl propionate hydrolysis in the presence of NADH and NADH and Mg2+ agree closely with the specific rates of acyl hydrolysis from the E . NADH . acyl and E . NADH . acyl . Mg2+ complexes seen in the dehydrogenase reaction with propionaldehyde. These observations support the view that esterase and dehydrogenase activities occur at the same site on the enzyme. Other evidence is presented to support this conclusion.


Bidirectional Mapping Perception-enhanced Cycle-consistent Generative Adversarial Network for Super-resolution of Brain MRI images.
As an effective tool for visualizing neurodegeneration, high-resolution structural magnetism facilitates quantitative image analysis and clinical applications. Super-resolution reconstruction technology allows to improve the resolution of images without upgrading the scanning hardware. However, existing super-resolution techniques relied on paired image data sets and lacked further quantitative analysis of the generated images. In this study, we proposed a semi-supervised generative adversarial network (GAN) model for super-resolution of brain MRI, and the synthetic images were evaluated using various quantitative measures. This model adopted the cycle-consistency structure to allow for a mixture of unpaired data for training. Perceptual loss was further introduced into the model to preserve detailed texture features at high frequencies. 363 subjects with both high-resolution (HR) and low-resolution (LR) scans and 217 subjects with HR scans only were used for model derivation, training, and validation. We extracted multiple voxel-based and surface-based morphological features of the synthetic and real 3D HR images for comparison. We further evaluated the synthetic images in the differential diagnosis of diseases. Our model achieved superior mean absolute error (0.049±0.021), mean squared error (0.0059±0.0043), peak signal-to-noise ratio (29.41±3.71), structural similarity index measure (0.914±0.048). Eight morphological metrics, both voxel-based and surface-based, showed significant agreement (P<0.0001). The gap of accuracy in disease diagnosis between synthetic and real HR images was within 5% and significantly outperformed the LR images. Our proposed model enables the reconstruction of HR MRI and could be used accurately for image quantification.Clinical relevance- Quantitative evaluation of the synthetic high-resolution images was used to determine whether the synthetic images have sufficient realism and diversity.


Postabsorptive and postprandial energy expenditure and substrate oxidation do not change during the menstrual cycle in young women.
There is little information on the effects of the menstrual cycle on the thermic effect of feeding (TEF) and postprandial substrate oxidation. To address these issues, a dose-response study was conducted in eight young women to whom test meals containing 0, 1046, 2092 and 4148 kJ were administered on nonconsecutive days, once each during the follicular phase and once during the luteal phase of the menstrual cycle. Postprandial energy expenditure was measured until it returned to base line (test duration for the 0 kJ meal was matched to duration of 4184 kJ meal). There was no significant difference between menstrual cycle phases in the duration of the thermogenic response to any size meal. TEF, calculated by four different methods, did not differ between follicular and luteal phases, and a comparable dose-response was observed in both phases. Similarly, there was no effect of menstrual cycle phase on postabsorptive or postprandial protein, fat, or carbohydrate oxidation. In conclusion, postabsorptive and postprandial energy expenditure and substrate oxidations did not differ in young women during the follicular and luteal phases of the menstrual cycle.


Development and clinical validation of an unobtrusive ambulatory knee function monitoring system with inertial 9DoF sensors.
Patients suffering from end-stage knee osteoarthritis are often treated with total knee arthroplasty, improving their functional mobility. A number of patients, however, report continued difficulty with stair ascent and descent or sportive activity after surgery and are not completely satisfied with the outcome. State-of-the-art analyses to evaluate the outcome and mobility after knee replacement are conducted under supervised settings in specialized gait labs and thus can only reflect a short period of time. A number of external factors may lead to artificial gait patterns in patients. Moreover, clinically relevant situations are difficult to simulate in a stationary gait lab. In contrast to this, inertial sensors may be used additionally for unobtrusive gait monitoring. However, recent notable approaches found in literature concerning knee function analysis have so far not been applied in a clinical context and have therefore not yet been validated in a clinical setting. The aim of this paper is to present a system for unsupervised long-term monitoring of human gait with a focus on knee joint function, which is applicable in patients' everyday lives and to report on the validation of this system gathered during walking with reference to state-of-the-art gait lab data using a vision system (VICON Motion System). The system KINEMATICWEAR - developed in close collaboration of computer scientists and physicians performing knee arthroplasty - consists of two sensor nodes with combined tri-axial accelerometer, gyroscope and magnetometer to be worn under normal trousers. Reliability of the system is shown in the results. An overall correlation of 0.99 (with an overall RMSE of 2.72) compared to the state-of-the-art reference system indicates a sound quality and a high degree of correspondence. KINEMATICWEAR enables ambulatory, unconstrained measurements of knee function outside a supervised lab inspection.


Health care in North must acknowledge Inuit values, traditional medicine.
Dr. Gail Gray spent a week on Baffin Island in a retreat devoted to discussions about health care delivery in the North. She says it is obvious that traditional medical practices and Inuit cultural values must be part of any new health care initiatives if the initiatives are to be successful.


History and evolution of methods for solving the inverse problem.
This article serves as an introduction to the other articles in this issue devoted to the problem of the localization of neural generators. Elements of the theory of electric volume conduction are briefly introduced, as far as these apply to the interpretation of observed scalp potentials. First, some basic methods for display of the different aspects of the spatiotemporal information are described. Next, the most prominent source and volume conductor models that have been postulated for the involved forward problem are summarized. The problems of source identification and source localization, known as the inverse problem, are then formulated in terms of a parameter estimation procedure. The importance of introducing a priori information in the inverse problem, aimed at stabilizing (regularizing) the obtained solution, is emphasized. Methods for imposing such constraints are briefly outlined.


Influence of nanoscale topology on bactericidal efficiency of black silicon surfaces.
The nanostructuring of materials to create bactericidal and antibiofouling surfaces presents an exciting alternative to common methods of preventing bacterial adhesion. The fabrication of synthetic bactericidal surfaces has been inspired by the anti-wetting and anti-biofouling properties of insect wings, and other topologies found in nature. Black silicon is one such synthetic surfaces which has established bactericidal properties. In this study we show that time-dependent plasma etching of silicon wafers using 15, 30, and 45 min etching intervals, is able to produce different surface geometries with linearly increasing heights of approximately 280, 430, and 610 nm, respectively. After incubation on these surfaces with Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa bacterial cells it was established that smaller, more densely packed pillars exhibited the greatest bactericidal activity with 85% and 89% inactivation of bacterial cells, respectively. The decrease in the pillar heights, pillar cap diameter and inter-pillar spacing corresponded to a subsequent decrease in the number of attached cells for both bacterial species.


Maximum-entropy models reveal the excitatory and inhibitory correlation structures in cortical neuronal activity.
Maximum entropy models can be inferred from large datasets to uncover how collective dynamics emerge from local interactions. Here, such models are employed to investigate neurons recorded by multi-electrode arrays in the human and monkey cortex. Taking advantage of the separation of excitatory and inhibitory neuron types, we construct a model including this distinction. This approach allows us to shed light on differences between excitatory and inhibitory activity across different brain states such as wakefulness and deep sleep, in agreement with previous findings. Additionally, maximum entropy models can also unveil novel features of neuronal interactions, which are found to be dominated by pairwise interactions during wakefulness, but are population-wide during deep sleep. Overall, we demonstrate that maximum entropy models can be useful to analyze datasets with classified neuron types and to reveal the respective roles of excitatory and inhibitory neurons in organizing coherent dynamics in the cerebral cortex.


BCL2 expression predicts survival in patients receiving synchronous chemoradiotherapy in advanced transitional cell carcinoma of the bladder.
It has been reported previously that BCL2 expression predicted a poor response to neo adjuvant chemotherapy but had no prognostic significance in patients receiving radiotherapy alone. We therefore investigated its role in patients receiving synchronous chemoradiotherapy as treatment for advanced bladder cancer. We examined expression of BCL2 and P53 by immunohistochemical analysis using archival tissue samples taken from patients included in the phase I/II trial of synchronous chemoradiotherapy for advanced transitional cell carcinoma (TCC) of the bladder. Data were collected on 24 patients who presented with invasive bladder cancer to the Queen Elizabeth Hospital between March 1998 and January 2001. Eleven patients had died at the time of analysis, with median follow-up of 34 months for the 13 surviving patients. Median survival for patients with strongly BCL2 positive tumours was 12.8 months while, for BCL2 weak or negative patients, the median is yet to be reached (p=0.03). The hazard ratio was 3.37 in favour of BCL2 negative tumours having longer survival. This study shows that over-expression of BCL2 in patients receiving synchronous chemoradiotherapy is an independent indicator of poor survival in muscle invasive TCC of the bladder.


Inorganic carbon utilization of tropical calcifying macroalgae and the impacts of intensive mariculture-derived coastal acidification on the physiological performance of the rhodolith Sporolithon sp.
Fish farming in coastal areas has become an important source of food to support the world's increasing population. However, intensive and unregulated mariculture activities have contributed to changing seawater carbonate chemistry through the production of high levels of respiratory CO2. This additional CO2, i.e. in addition to atmospheric inputs, intensifies the effects of global ocean acidification resulting in localized extreme low pH levels. Marine calcifying macroalgae are susceptible to such changes due to their CaCO3 skeleton. Their physiological response to CO2-driven acidification is dependent on their carbon physiology. In this study, we used the pH drift experiment to determine the capability of 9 calcifying macroalgae to use one or more inorganic carbon (Ci) species. From the 9 species, we selected the rhodolith Sporolithon sp. as a model organism to investigate the long-term effects of extreme low pH on the physiology and biochemistry of calcifying macroalgae. Samples were incubated under two pH treatments (pH 7.9 = ambient and pH 7.5 = extreme acidification) in a temperature-controlled (26 ± 0.02 °C) room provided with saturating light intensity (98.3 ± 2.50 μmol photons m-2 s-1). After the experimental treatment period (40 d), growth rate, calcification rate, nutrient uptake rate, organic content, skeletal CO3-2, pigments, and tissue C, N and P of Sporolithon samples were compared. The pH drift experiment revealed species-specific Ci use mechanisms, even between congenerics, among tropical calcifying macroalgae. Furthermore, long-term extreme low pH significantly reduced the growth rate, calcification rate and skeletal CO3-2 content by 79%, 66% and 18%, respectively. On the other hand, nutrient uptake rates, organic matter, pigments and tissue C, N and P were not affected by the low pH treatments. Our results suggest that the rhodolith Sporolithon sp. is susceptible to the negative effects of extreme low pH resulting from intensive mariculture-driven coastal acidification.


Nanoparticle-based immunoengineering strategies for enhancing cancer immunotherapy.
Cancer immunotherapy is a groundbreaking strategy that has revolutionized the field of oncology compared to other therapeutic strategies, such as surgery, chemotherapy, or radiotherapy. However, cancer complexity, tumor heterogeneity, and immune escape have become the main hurdles to the clinical application of immunotherapy. Moreover, conventional immunotherapies cause many harmful side effects owing to hyperreactivity in patients, long treatment durations and expensive cost. Nanotechnology is considered a transformative approach that enhances the potency of immunotherapy by capitalizing on the superior physicochemical properties of nanocarriers, creating highly targeted tissue delivery systems. These advantageous features include a substantial specific surface area, which enhances the interaction with the immune system. In addition, the capability to finely modify surface chemistry enables the achievement of controlled and sustained release properties. These advances have significantly increased the potential of immunotherapy, making it more powerful than ever before. In this review, we introduce recent nanocarriers for application in cancer immunotherapy based on strategies that target different main immune cells, including T cells, dendritic cells, natural killer cells, and tumor-associated macrophages. We also provide an overview of the role and significance of nanotechnology in cancer immunotherapy.


A preliminary crystallographic investigation of avidin.
Avidin was crystallized in various forms from 3m-ammonium sulphate or sodium phosphate at pH5.2. X-ray-crystallographic analysis of crystals from phosphate showed them to be orthorhombic, space group C222, containing two molecules per unit cell and with a single subunit as the asymmetric unit. The formation of the complex with biotin did not affect the morphology of the crystals.


Outcome of single-stage versus two-stage exchange for revision knee arthroplasty for chronic periprosthetic infection.
The gold standard for treating chronic periprosthetic joint infection is still considered to be double-stage exchange revision. The purpose of this review is to analyse the difference in terms of eradication rates and functional outcome after single- and double-stage prosthetic exchange for chronic periprosthetic joint infection around the knee.We reviewed full text articles written in English from 1992 to 2018 reporting the success rates and functional outcomes of either single-stage exchange or double-stage exchange for knee arthroplasty revision performed for chronic infection. In the case of double-stage exchange, particular attention was paid to the type of spacer: articulating or static.In all, 32 articles were analysed: 14 articles for single-stage including 687 patients and 18 articles for double-stage including 1086 patients. The average eradication rate was 87.1% for the one-stage procedure and 84.8% for the two-stage procedure. The functional outcomes were similar in both groups: the average Knee Society Knee Score was 80.0 in the single-stage exchange group and 77.8 in the double-stage exchange. The average range of motion was 91.4° in the single-stage exchange group and 97.8° in the double-stage exchange group.Single-stage exchange appears to be a viable alternative to two -stage exchange in cases of chronic periprosthetic joint infection around the knee, provided there are no contra-indications, producing similar results in terms of eradication rates and functional outcomes, and offering the advantage of a unique surgical procedure, lower morbidity and reduced costs. Cite this article: EFORT Open Rev 2019;4:495-502. DOI: 10.1302/2058-5241.4.190003.


ANSWER: Annotation Software for Electronic Reporting.
To better use genetic testing, which is used by clinicians to explain the molecular mechanism of disease and to suggest clinical actionability and new treatment options, clinical next-generation sequencing (NGS) laboratories must send the results into reports in PDF and discrete data element format (HL7). Although most clinical diagnostic tests have set molecular markers tested and have a set range of values or a binary result (positive or negative), the NGS genetic test could examine hundreds or thousands of genes with no predefined list of variants. Although there are some commercial and open-source tools for clinically reporting genomics results for oncology testing, they often lack necessary features. Using several available software tools for data storage including MySQL and MongoDB, database querying with Python, and a web-based user application using JAVA and JAVA script, we have developed a tool to store and query complex genomics and demographics data, which can be manually curated and reported by the user. We have developed a tool, Annotation SoftWare for Electronic Reporting (ANSWER), that can allow molecular pathologists to (1) filter variants to find those meeting quality control metrics in the genes that are clinically actionable by diagnosis; (2) visualize variants using data generated in the bioinformatics analysis; (3) create annotations that can be reused in future reports with association specific to the gene, variant, or diagnosis; (4) select variants and annotations that should be reported to match the details of the case; and (5) generate a report that includes demographics, reported variants, clinical actionability annotation, and references that can be exported into PDF or HL7 format, which can be electronically sent to an electronic health record. ANSWER is a tool that can be installed locally and is designed to meet the clinical reporting needs of a clinical oncology NGS laboratory for reporting.


Transcriptional regulation of Rab32/38, a specific marker of pigment cell formation in Ciona robusta.
Through a myriad of pigments stored in different cells, animal pigmentation represents a crucial process to face disparate environmental and ecological challenges. In vertebrates, the small GTPase Rab32 and Rab38 have a conserved role in the transport of key melanogenic enzymes, as tyrosinase (tyr) and tyrosinase-related protein (tyrp), to the melanosomes in formation. We provide a survey on Rab32/38 evolution and its regulatory logics during pigment cell formation in Ciona robusta. Our phylogeny supports the existence of a single Rab32/38 gene in tunicates, which is probably the unique transporter for tyrosinase family members in this clade. Different deletions allow us to identify the minimal cis-regulatory element able to recapitulate the endogenous gene expression during pigment cell development in C. robusta. In this conserved region, we identified two putative binding sites for the transcription factor Mitf, which is known for its role as regulator of pigmentation in vertebrates. Mutational analysis revealed that both Mitf binding sites are essential for the activity of this regulatory region and we demonstrated that Mitf misexpression is able to induce ectopic activation of the Rab32/38 regulatory region in vivo. Our results strongly indicate that Mitf is involved in the regulation of Rab32/38 activity during Ciona pigment cell development.


Clinical outcomes of hemiarthroplasty and biological resurfacing in patients aged younger than 50 years.
Total shoulder arthroplasty as a treatment for glenohumeral degenerative joint disease is well accepted but has been less predictable with regard to outcomes and durability in a younger aged population, typically aged younger than 50 years. This younger population has a greater potential for glenoid component loosening. This has led surgeons to perform hemiarthroplasty or hemiarthroplasty with biological resurfacing of the glenoid in an effort to avoid the potential problems with a polyethylene glenoid and obtain durable and acceptable results for these patients. The study included 44 patients, with 23 undergoing hemiarthroplasty alone and 21 undergoing hemiarthroplasty with biological resurfacing of the glenoid. All patients were aged younger than 50 years. Preoperative diagnoses, comorbidities, demographics, and range of motion were collected. Preoperative and postoperative radiographs were obtained. Preoperative and postoperative objective scoring measures (Single Assessment Numeric Evaluation, American Shoulder and Elbow Surgeons score, visual analog scale, Simple Shoulder Test, Constant-Murley) were used. Mean follow-up was 3.8 years for the hemiarthroplasty group and 3.6 years for the biological resurfacing group. Six patients in the hemiarthroplasty and 12 patients in the biological resurfacing group were considered failures due to revision surgery or an American Shoulder and Elbow Surgeons score <50. The hemiarthroplasty group had significantly better visual analog scale and Single Assessment Numeric Evaluation scores. There was a significant failure rate in the hemiarthroplasty and the biologic resurfacing groups compared with results in the literature. Improved outcomes and lower failure rates were observed in the hemiarthroplasty group compared with the biological resurfacing group in this study.


Heavy chain variable region. Multiple gene segments encode anti-4-(hydroxy-3-nitro-phenyl)acetyl idiotypic antibodies.
The hapten (4-hydroxy-3-nitrophenyl)acetyl (NP), when conjugated to carrier proteins, elicits a characteristic idiotypic response (NPb) in C57BL/6 mice. The response can be divided serologically into two distinct NPb-positive groups of antibodies. The first group consists of four crossreacting subgroups (I-IV), the second of two subgroups (V, VI). Some antibodies of subgroups I and II have been shown to express the unmutated heavy chain variable region (VH) germline gene 186.2. Antibodies of subgroups V and VI crossreact extensively with the NPa-positive antibodies of BALB/c mice. We sequenced heavy chain complementary DNA from eight hybridomas producing anti-NP antibodies. Six of these belong to subgroups V and VI, and two were NPa-positive hybridomas of BALB/c origin. All sequences were homologous to each other, and differed by approximately 80 basepairs from the 186.2 C57BL/6 germline VH gene. From our sequence and Southern blot analyses we suggest: (a) the NPb idiotypic response is the product of several VH germline genes, (b) some of these genes are very homologous to the gene coding for the BALB/c NPa idiotype, and might represent the C57BL/6 allelic forms of this gene, (c) the diversity regions of NPb and NPa-positive antibodies are diverse in length and amino acid composition, except for the first residue, which is always tyrosine, (d) all four heavy chain joining region gene segments are expressed without mutation. We discuss our data in terms of diversity in the germline VH gene repertoire, as well as diversity created by gene segment-joining events and somatic mutation.


Association of chronic kidney disease with cardiovascular disease in cancer patients: a cross-sectional study.
Due to the cardiotoxicity of cancer treatment and traditional risk factors for cardiovascular disease (CVD) such as obesity, diabetes, dyslipidemia, and hypertension, cancer patients are at higher risk of developing CVD. However, limited research exists on the correlation between chronic kidney disease (CKD) and CVD risk in cancer patients. This cross-sectional study selected cancer patients aged ≥20 years from the National Health and Nutrition Examination Survey (NHANES) conducted from 2015 to 2020. Multivariable logistic regression was used to assess the association between CKD and CVD in cancer patients. Additionally, subgroup analyses were conducted to investigate the association among different groups of cancer patients. We included 1700 adult cancer patients (52.53% were female). After multivariable adjustment for covariates including traditional CVD factors, CKD was significantly associated with CVD, with an odds ratio (95% confidence interval) and P-value of 1.61(1.18,2.19) and 0.004. Subgroup analyses after multivariable adjustment showed a significant correlation between CKD and increased CVD risk in the following populations: age ≥60 years, males, White ethnicity, and individuals with or without traditional CVD factors (obesity, diabetes, dyslipidemia, and hypertension). CKD remains a significant factor in the higher risk of CVD among adult cancer patients in the United States, even after adjustment for traditional CVD risk factors. Therefore, to reduce the risk of CVD in cancer patients, it is important to treat CKD as a non-traditional risk factor for CVD and actively manage it.


[Oscilloscopic control of the microvibration occurring in the preparation of sections for electron microscopic study].
For purposes of control and timely elimination of microvibration arising during the cutting of ultrathin tissue slices, a method is proposed using the piezocrystal, which is secured to the epoxy block with the help of melted paraffin. The elimination of microvibration is carried out by changing the angle of inclination or the speed of cutting in a broad range from 1 to 15 degrees, and from 0.5 to 5.0 cm/sec, resp. The noise and vibration outside the ultratome does not induce microvibration during the ultrathin slice preparation.


Coffee consumption and risk of coronary heart diseases: a meta-analysis of 21 prospective cohort studies.
A large amount of cohort studies addressed coffee consumption and risk of coronary heart disease (CHD) and yielded inconsistent results. We conducted a meta-analysis to estimate the pooling effects. We searched for all published English articles indexed in MEDLINE or PubMed from January 1966 to January 2008. Twenty-one independent prospective cohort studies, which tested CHD risk by coffee consumption, were identified. A general variance-based method was used to pool the relative risks (RR). 15,599 cases from 407,806 participants were included in pooling the overall effects. As compared to the light coffee consumption (<1 cup/d in US or <or=2 cups/d in Europe), under the random-effects model, the pooled CHD RRs (95% CI) for all studies combined were 0.96 (0.87-1.06), 1.04 (0.92-1.17) and 1.07 (0.87-1.32) for the moderate (1-3 or 3-4 cups/d), heavy (4-5 or 5-6 cups/d) and very heavy (>or=6 or >or=7 cups/d) categories of coffee consumption (all p>0.05); Moderate coffee consumption showed significantly lower CHD RR (95% CI) of 0.82 (0.73-0.92) (p<0.001) in women, and of 0.87 (0.80-0.86) (p=0.001) in men and women followed <or=10 years. Our findings do not support the hypothesis that coffee consumption increases the long-term risk of coronary heart disease. Habitual moderate coffee drinking was associated with a lower risk of CHD in women.


Temporal lobe epilepsy and aggression.
Research on temporal lobe epilepsy (TLE) and aggression is reviewed in order to learn why it has not contributed more to our understanding of the neural basis of aggression. The research problem can be resolved into two questions: Are temporal lobe epileptics more aggressive? What inferences can be drawn about the factors underlying aggression in TLE? Difficulties in finding suitable operational definitions for TLE and for aggression have, to date, received insufficient attention. In published studies, sample bias and lack of regard for the validity and reliability of behavioral assessments prevent our deciding whether an association exists between interictal aggressive behavior and TLE. Even if this association were demonstrated, one could not draw direct neurobehavioral inferences from it, since a variety of social and psychological, as well as neurophysiological variables could contribute to the association. These would have to be controlled. Methodological refinements could lead to more satisfactory answers to the first question, but the second is considerably more difficult. Some sources of complexity and apparent contradictions in the experimental literature on the neurology of aggression are discussed. They consist of technical problems, the complexity of the neural substrate of aggression and the influence of environmental cues and learning. The combined use of electrophysiological and interview techniques has yielded some interesting results with TLE patients.


Assessing stakeholders' opinions on sharing antimicrobial susceptibility testing data from animals is key to developing a centralized database and dashboard tool.
To present, analyze, and discuss stakeholders' opinions regarding sharing antimicrobial susceptibility testing (AST) data from animals into a centralized database and dashboard tool that would collect, aggregate, store, and analyze this type of data from veterinary diagnostic laboratories (VDLs) across the country. 1 in-person focus group (9 participants), 9 virtual focus groups (49 participants), and online pre- and postmeeting surveys (76 and 35 participants, respectively). Focus groups and surveys were conducted to assess the opinions of veterinarians, producers, researchers, diagnosticians, and government officials. A strong majority of stakeholders recognize AMR as a serious concern for both human and animal health and see several benefits in establishing a centralized AMR database; however, several concerns were raised associated with data confidentiality, security, curation, and harmonization. In the interest of alleviating those concerns, among other items, stakeholders suggested education and training of data users, providers, and the public in addition to assuring strong data confidentiality protections. Stakeholder engagement is a critical component of all stages of development, implementation, and utilization of an AMR database and dashboard tool that could be used to inform antimicrobial stewardship in veterinary medicine. This assessment of stakeholders' needs and concerns can be used to help guide future recommendations for data legal protections as well as a data confidentiality and security framework. Maintaining open communication on data usage, storage, and security as well as involvement and education of data providers, users, and the public will remain key to enabling development of an AMR database and dashboard tool for domesticated animals.


Effects of PMA (PHORBOL-12-MYRISTATE-13-ACETATE) on the Developing Rodent Brain.
Perinatal infections have a negative impact on brain development. However, the underlying mechanisms leading to neurological impairment are not completely understood and reliable models of inflammation are urgently needed. Using phorbol-myristate-acetate as an activator of inflammation, we investigated the effect on the developing rodent brain. Neonatal rats and mice deficient in IL-18 or IRAK-4 were exposed to PMA. Brains were assessed for regulation of pro- and anti-inflammatory cytokines and cell death 24 hrs, 7 and 14 days after treatment. PMA induced an inflammatory response and caused widespread neurodegeneration in the brains of 3- and 7-day-old rats. In contrast, 14-day-old rats were resistant to the neurotoxic effect of PMA. Histological evaluation at the age of 14 and 21 days revealed a destruction of the cortical microstructure with decreased numerical density of neuronal cells. Mice deficient in IL-18 or IRAK-4 were protected against PMA induced brain injury. PMA treatment during a vulnerable period can alter brain development. IL-18 and IRAK-4 appear to be important for the development of PMA induced injury.


Takotsubo Cardiomyopathy in a Patient with Undiscovered Sigmoid Colon Cancer.
Takotsubo cardiomyopathy (TTC) is a stress-related cardiomyopathy that is characterized by reversible left systolic dysfunction, which appears to be precipitated by sudden emotional or physical stress in the absence of myocardial infarction. Here we present a rare case that clinically presented with intermittent abdominal pain, initially impressed as non-ST elevation myocardial infarction and congestive heart failure but with a normal coronary angiogram. Her symptoms relieved spontaneously without returning. Sigmoid colon cancer was diagnosed via colonoscopy later due to persistent abdominal discomfort. In the absence of detectable emotional or physical stress factors, the newly diagnosed sigmoid colon cancer was the only possible trigger factor of TTC. We offer this case as a reminder that cancer should be considered in the differential diagnosis of patients presenting with the etiology of TTC.


Depressed interferon synthesis in skin fibroblasts from lung cancer patients.
Skin fibroblast cell cultures, derived from male adult lung cancer patients, an adult control population, and a newborn population were examined for their susceptibility to transformation with Kirsten murine sarcoma virus and their ability to respond to an interferon inducer (poly I X poly C). An association between sensitivity to viral transformation and induction of interferon was observed. Cultures derived from lung cancer patients demonstrated an increased sensitivity to virus transformation and a decreased ability to respond to interferon induction as compared with age-matched controls and newborns.


Review of the treatment of mycosis fungoides and Sézary syndrome: A stage-based approach.
Mycosis fungoides (MF) and Sézary Syndrome (SS) are the most common subtypes of cutaneous T-cell lymphomas. Most of patients have indolent and incurable course of disease. Therefore, treatment should be reaching the optimal benefit with minimizing the toxicity as much as possible. To achieve this aim, the management should follow a -stage-based-approach. Treatment of early-stage MF (IA-IIA) involves skin-directed therapy (SDT) including topical corticosteroids, phototherapy, topical chemotherapy, topical retinoids and radiotherapy. For aggressive/recalcitrant early-stage MF or advanced-stage MF, systemic therapy should be considered including interferone-alpha, oral retinoids including bexarotine and more recently acitretin, histone deacetylase inhibitors (HDACi), fusion toxin denileukin diftitox and chemotherapy drugs. Combined drug regimens can be considered in some situations to get the synergistic effect while lowering the individual drug's doses on the other hand. By exception of aggressive stages, chemotherapy should always come after other systemic drugs have been tried or contraindicated. Novel drugs should be considered in situations when all systemic drugs have failed.


Treating respiratory infections in the era of cost control.
The treatment of lower respiratory tract infections will undoubtedly be affected by cost-control measures. Probably the most important change in the diagnosis and treatment of bronchitis and pneumonia will be a more empiric approach. A basic knowledge of likely pathogens, plus the clinical judgment to provide treatment modifications for patients susceptible to unusual pathogens, will usually assure a successful outcome.