Tumor-specific idiotype vaccines. I. Generation and characterization of internal image tumor antigen.
The concept of idiotype vaccines against tumor-associated antigens (TAA) was tested in the DBA/2 L1210 lymphoma subline, L1210/GZL. Monoclonal antibodies against a TAA that cross-reacts with the envelope glycoprotein gp52 of the mammary tumor virus were used to make hybridoma anti-idiotype antibodies (Ab2). In this report we describe the characterization of monoclonal anti-idiotypic antibodies against the combining site of 11C1 (Ab1), which recognizes a shared determinant of gp52 of mouse mammary tumor virus (MMTV) and the TAA of L1210/GZL. Hybridomas expressing the internal image of gp52 were screened by an idiotype inhibition assay. Mice sensitized with radiated L1210/GZL cells produced specific delayed type hypersensitivity (DTH) against the Ab2 hybridoma. Five Ab2 hybridomas were selected and were used to immunize DBA/2 mice. Such immunized animals showed specific DTH reaction against a challenge with the L1210/GZL tumor cells. Similar results were obtained in mice immunized with purified Ab2. Fluorescence-activated cell sorter analysis demonstrated that fluorescence staining of L1210/GZL cells by 11C1 can be completely inhibited with preabsorption on Ab2 hybridoma cells. Mice immunized with 2F10 and 3A4 coupled to keyhole limpet hemocyanin (KLH) contained antibodies binding to MMTV. But only in mice immunized with 2F10-KLH was significant inhibition of L1210/GZL tumor growth observed. Collectively, these results indicate that certain anti-idiotypic antibodies can mimic the MMTV gp52 antigen, as well as the gp52-like epitope expressed on the L1210/GZL tumor cells. These properties of anti-idiotypic antibodies mimicking TAA could be exploited for making idiotype vaccines against tumors.


Losing it.
"It's worse than I thought.... She's completely lost her mind," says Harry Beecham, the CEO of blue chip management consultancy Pierce and Company. The perplexed executive was in a hotel suite with his wife in Amsterdam, the latest stop on his regular trek to dozens of Pierce offices worldwide. In his hand was a sheaf of paper--the same message sent over and over again by his star employee and protégée Katharina Waldburg. The end of the world is coming, she warned. "Someone is going to die." Harry wouldn't have expected this sort of behavior from Katharina. After graduating with distinction from Oxford, she made a name for herself by single-handedly building Pierce's organizational behavior practice. At 27, she's poised to become the youngest partner ever elected at the firm. But Harry can't ignore the faxes in his hand. Or the stream-of-consciousness e-mails Katharina's been sending to one of the directors in Pierce's Berlin office--mostly gibberish but potentially disastrous to Katharina's reputation if they ever got out. Harry also can't dismiss reports from Roland Fuoroli, manager of the Berlin office, of a vicious verbal exchange Katharina had with him, or of an "over the top" lunch date Katharina had with one of Pierce's clients in which she was explaining the alphabet's role in the creation of the universe. Harry is planning to talk to Katharina when he gets to Berlin. What should he say? And will it be too late? Four commentators offer their advice in this fictional case study. They are Kay Redfield Jamison, a professor of psychiatry and a coauthor of Manic-Depressive Illness; David E. Meen, a former director at McKinsey & Company; Norman Pearlstine, the editor in chief at Time Incorporated; and Richard Primus, an assistant law professor at the University of Michigan.


Hospital middle management experience and peer interpersonal conflict.
Each manager brings a unique decision style to the problem-solving process. This decision style may be quite different from the decision style of other managers and a potential source of interpersonal conflict. A study was undertaken to determine which decision styles were potential sources of interpersonal conflict for hospital middle managers and to determine whether years of management experience had any bearing on the sources of conflict. Study results reported interpersonal conflict with decision styles that were impersonal, logical, and factual. Results were consistent regardless of years of management experience.


Peptide inhibitors against influenza virus.
Influenza A virus is a particularly problematic virus because of its ability to cause high levels of morbidity on a global scale within a remarkably short period of time. It also has the potential to kill very large numbers of people as occurred in the Spanish influenza pandemic in 1918. Options for antiviral therapy are limited because of the paucity of available drugs and the rapid mutation rate of the virus leading to the emergence of drug-resistant strains. The current H1N1 pandemic and potential threats posed by other strains highlight the need to develop novel therapeutic and prophylactic strategies. Here, we summarize the current state and recent developments of peptide-based inhibitors of influenza A virus.


[Investigation of ribosomal RNAs from liver mitochondria of albino rats].
Study of sedimentation coefficients of rat liver mt-rRNAs labelled in vivo under conditions of selective inhibition by actinomycin D of nuclear RNAs biosynthesis, in hypoosmic system of isolated mitochondria, and of mt-rRNAs isolated from purified 55 ribosomes revealed that mt-rRNAs are represented by 16S and 12S components. Both components are found to be of AU-type. Anomalous behaviour of mtRNA on MAK columns is observed. Some aspects of isolation and identification of mt-rRNAs in higher organisms are discussed.


Human papillomavirus in clinically and histologically normal tissue of patients with genital cancer.
To study the association of human papillomavirus (HPV) and herpes simplex virus (HSV) with genital cancer, we collected specimens of cervical, vulvar, endometrial, and vaginal tumors at the time of operation in patients with cancer. In some patients, matched internal-control (histologically normal) tissue was also collected. DNA extracted from the tissue was probed with radiolabeled HPV type 16 DNA, HPV type 18 DNA, and cloned fragments of HSV type 2 DNA. Hybridization to the HindIIIa clone of HSV-2 was detected in only 1 cervical tumor and 1 vulvar tumor (9 percent) among the 22 tumors tested. However, DNA sequences hybridizing to HPV-16 were detected in 21 of 25 tumors (84 percent) and in 8 of 11 (73 percent) of the DNA samples from clinically and histologically normal, paired, internal-control tissues from the patients with cancer. HPV-16 DNA was found in one of nine normal cervixes (11 percent) of women without genital neoplastic disease or abnormal cytology. HPV-18 DNA was detected in only 2 of 24 tumors (8 percent), 1 cervical and 1 vulvar. Our results show a strong association between the presence of HPV-16 genomes and genital tumors and between HPV-16 genomes and histologically normal tissue within 2 to 5 cm of the tumors. The implications of these findings remain to be explored.


[Diagnostic of gestational diabetes mellitus and the prevalence of LGA (Large for Gestational Age)].
Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance, first time detected in pregnancy. Early diagnosis of the disease may reduce fetal exposure to maternal hyperglycemia and decrease the risk of LGA. The aim of the study was to examine the influence of time and diagnostic method of GDM on the prevalence of LGA and pregnancy outcome among patients with gestational diabetes. The study was conducted among 211 women with gestational diabetes mellitus, the patients of 1st Clinic of Obstetrics and Gynecology at the Medical University in Warsaw. We have reviewed the results of fasting plasma glucose, 50-g glucose screening test (GCT) and 2 hour 75-g glucose tolerance test in GDM patients with LGA and eutrophic newborns. The t-student or the Mann-Whitney test was used in order to compare both groups. P<0.05 was deemed statistically significant. LGA was diagnosed in 10.4% of patients. We did not find any significant differences in gestational age when GDM was diagnosed, results of fasting glucose GCT and OGTT among LGA (M) and control (K) group. However, when we compared the percentage of LGA in groups of women with different time of GDM diagnosis, the highest prevalence was noted in the group of first trimester diagnosis and between 28 and 32 weeks of pregnancy, which we found interesting. We compared the women and the results of the diagnostic tests with the group of standard time of GDM diagnosis (24-28 week of pregnancy) and the only difference was the late diagnosis. If 75-g glucose tolerance test had not included 1-hour after load glucose assignment, GDM would not have been diagnosed at all in 18.2% of female patients with LGA. We have not found any correlations between the results of the diagnostic tests, the time of the diagnosis or the mode of treatment GDM (diet alone or with insulin) and the birth weight. 1. Results of fasting glucose and glucose tolerance tests are not useful in the prediction of LGA in GDM pregnancies. 2. Diagnosis of GDM during the recommended period (between 24 and 28 weeks of pregnancy) may decrease the prevalence of LGA (comparing to later diagnostics). 3. 75-g glucose tolerance test should provide fasting, 1 and 2-hour after load glucose assignment.


Isolable rubidium and caesium derivatives of common organic carbonyl compounds.
Light alkali metal (Li, Na, K) amides have a long history of synthetic utility, but heavier (Rb, Cs) congeners have barely been studied. This study reveals remarkable structurally complex outcomes of reacting AM(HMDS) (AM = Rb, Cs; HMDS = hexamethyldisilazide) with benzaldehyde and acetophenone. Though complicated, reactions give a diversity of eye-catching isolated products, an enolate with a hexagonal prismatic network, two dienolates with distinct extended ladder motifs, and two β-imino-alkoxides comprising zig-zag chains of metal-oxygen bonds in infinite cages.


[Paraneoplastic syndrome].
Treatment for the paraneoplastic syndrome associated with lung cancer was reviewed. The principle of the treatment of paraneoplastic syndrome is to control cancer as an underlying disease. Therefore, the standard therapy for Cushing's syndrome associated with lung cancer is surgical treatment if the tumor is operable. There is no standard therapy for Cushing's syndrome associated with advanced small-cell lung cancer. Metyrapone is used in combination with systemic chemotherapy. The effects of ketoconazole and octreotide are under investigation. To control hyponatremia due to the syndrome of inappropriate antidiuretic hormone secretion, fluid restriction is standard. When hyponatremia cannot be controlled with fluid restriction, demeclocycline can be used. For life-threatening hyponatremia, hypertonic saline with intravenous furosemide is administered under careful monitoring. Followed by hydration with saline, pamidronate is effective for the control of symptomatic hypercalcemia. Combined use of calcitonin facilitates rapid normalization of serum calcium for critically ill cases. Heparin is used for patients with recurrent episodes of thrombosis resulting from chronic disseminated intravascular coagulation, although the efficacy is controversial. Thrombocytes and coagulation factors are combined with heparin for patients with uncontrollable bleeding, although the efficacy is not established.


Development of a new psychophysical method to assess intranasal trigeminal chemosensory function.
The aim of this study was to develop a new psychophysical test to assess intranasal trigeminal chemosensory function. The test is similar to the Sniffin' Sticks test, but using pens impregnated with substances preferentially activating trigeminal afferents. Our test comprises detection threshold, discrimination, identification and lateralization tasks. In a first study, we evaluated healthy controls. In a second study, we evaluated the potential usefulness of this test in patients with rhinological conditions. Study 1: 86 controls were included. Threshold, identification and lateralization performance decreased with age. Test-retest reliability was similar to that of olfactory tests. Study 2: results of the controls group were compared to those of 59 patients (14 allergic rhinitis, 11 chronic rhinosinusitis with nasal polyps (CRSwNP), 9 without nasal polyps (CRSsNP), and 25 with an olfactory disorder (OD)). Controls had 1) lower detection thresholds compared to CRSwNP, CRSsNP and OD, 2) better discrimination and identification scores compared to OD, and 3) better lateralization scores compared to CRSwNP and CRSsNP. Our test allows to identify age-related changes in trigeminal chemosensory function. Trigeminal function seems to be differently affected in different pathologies. Further studies are necessary to validate our results and evaluate the impact of olfactory co-activation on the observed results.


In Situ Photochemical Activation of Sulfate for Enhanced Degradation of Organic Pollutants in Water.
The advanced oxidation process (AOP) based on SO4•- radicals has been receiving growing attention in water and wastewater treatment. Producing SO4•- radicals by activation of peroxymonosulfate or persulfate faces the challenges of high operational cost and potential secondary pollution. In this study, we report the in situ photochemical activation of sulfate (i-PCAS) to produce SO4•- radicals with bismuth phosphate (BPO) serving as photocatalyst. The prepared BPO rod-like material could achieve remarkably enhanced degradation of 2,4-dichlorophenol (2,4-DCP) in the presence of sulfate, indicated by the first-order kinetic constant (k = 0.0402 min-1) being approximately 2.1 times that in the absence (k = 0.019 min-1) at pH-neutral condition. This presented a marked contrast with commercial TiO2 (P25), the performance of which was always inhibited by sulfate. The impact of radical scavenger and electrolyte, combined with electron spin resonance (ESR) measurement, verified the formation of •OH and SO4•- radicals during i-PCAS process. According to theoretical calculations, BPO has a sufficiently high valence band potential making it thermodynamically favorable for sulfate oxidation, and weaker interaction with SO4•- radicals resulting in higher reactivity toward target organic pollutant. The concept of i-PCAS appears to be attractive for creating new photochemical systems where in situ production of SO4•- radicals can be realized by using sulfate originally existing in aqueous environment. This eliminates the need for extrinsic chemicals and pH adjustment, which makes water treatment much easier, more economical, and more sustainable.


Minireview: demystifying microbial reaction energetics.
The biology literature is rife with misleading information on how to quantify catabolic reaction energetics. The principal misconception is that the sign and value of the standard Gibbs energy ( Δ G r 0 ) define the direction and energy yield of a reaction; they do not. Δ G r 0 is one part of the actual Gibbs energy of a reaction (ΔGr ), with a second part accounting for deviations from the standard composition. It is also frequently assumed that Δ G r 0 applies only to 25 °C and 1 bar; it does not. Δ G r 0 is a function of temperature and pressure. Here, we review how to determine ΔGr as a function of temperature, pressure and chemical composition for microbial catabolic reactions, including a discussion of the effects of ionic strength on ΔGr and highlighting the large effects when multi-valent ions are part of the reaction. We also calculate ΔGr for five example catabolisms at specific environmental conditions: aerobic respiration of glucose in freshwater, anaerobic respiration of acetate in marine sediment, hydrogenotrophic methanogenesis in a laboratory batch reactor, anaerobic ammonia oxidation in a wastewater reactor and aerobic pyrite oxidation in acid mine drainage. These examples serve as templates to determine the energy yields of other catabolic reactions at environmentally relevant conditions.


Characteristics of physiological inducers of the ethanol utilization (alc) pathway in Aspergillus nidulans.
The ethanol utilization (alc) pathway in Aspergillus nidulans is one of the strongest expressed gene systems in filamentous fungi. The pathway-specific activator AlcR requires the presence of an inducing compound to activate transcription of genes under its control. We have demonstrated recently that acetaldehyde is the sole physiological inducer of ethanol catabolism. In the present study we show that compounds with catabolism related to that of ethanol, i.e. primary alcohols, primary monoamines and l-threonine, act as inducers because their breakdown results in the production of inducing aliphatic aldehydes. Such aldehydes were shown to induce the alc genes efficiently at low external concentrations. When ethanol is mixed with representatives of another class of strong direct inducers, ketones, the physiological inducer, acetaldehyde, prevails as effector. Although direct inducers essentially carry a carbonyl function, not all aldehydes and ketones act as inducers. Structural features discriminating non-inducing from inducing compounds concern: (i) the length of the aliphatic side group(s); (ii) the presence and nature of any non-aliphatic substituent. These characteristics enable us to predict whether or not a given carbonyl compound will induce the alc genes.


Convergence of inhibitory neural inputs regulate motor activity in the murine and monkey stomach.
Inhibitory motor neurons regulate several gastric motility patterns including receptive relaxation, gastric peristaltic motor patterns, and pyloric sphincter opening. Nitric oxide (NO) and purines have been identified as likely candidates that mediate inhibitory neural responses. However, the contribution from each neurotransmitter has received little attention in the distal stomach. The aims of this study were to identify the roles played by NO and purines in inhibitory motor responses in the antrums of mice and monkeys. By using wild-type mice and mutants with genetically deleted neural nitric oxide synthase (Nos1-/-) and P2Y1 receptors (P2ry1-/-) we examined the roles of NO and purines in postjunctional inhibitory responses in the distal stomach and compared these responses to those in primate stomach. Activation of inhibitory motor nerves using electrical field stimulation (EFS) produced frequency-dependent inhibitory junction potentials (IJPs) that produced muscle relaxations in both species. Stimulation of inhibitory nerves during slow waves terminated pacemaker events and associated contractions. In Nos1-/- mice IJPs and relaxations persisted whereas in P2ry1-/- mice IJPs were absent but relaxations persisted. In the gastric antrum of the non-human primate model Macaca fascicularis, similar NO and purine neural components contributed to inhibition of gastric motor activity. These data support a role of convergent inhibitory neural responses in the regulation of gastric motor activity across diverse species.


The partition and transport behavior of cytotoxic ionic liquids (ILs) through the DPPC bilayer: Insights from molecular dynamics simulation.
A molecular dynamics (MD) simulation with atomistic details was performed to examine the partitioning and transport behavior of moderately cytotoxic ionic liquids (ILs), namely choline bis(2-ethylhexyl) phosphate (CBEH), choline bis(2,4,4-trimethylpentyl) phosphinate (CTMP) and choline O,O-diethyl dithiophosphate (CDEP) in a fully hydrated dipalmitoylphosphatidylcholine (DPPC) bilayer in the fluid phase at 323 K. The structure of ILs was so selected to understand if the role of dipole and dispersion forces in the ILs distribution in the membrane can be possible. Several analyses including mass density, electrostatic potential, order parameter, diffusion coefficients and hydrogen bond formation, was carried out to determine the precise location of the anionic species inside the membrane. Moreover, the potential of the mean force (PMF) method was used to calculate free energy profile for transferring anionic species from the DPPC membrane into the bulk water. While less cytotoxic DEP is located within the bulk water, more cytotoxic TMP and BEH ILs were found to remain in the membrane and the energy barrier for crossing through the bilayer center of BEH was higher. Various ILs have no significant effect on P-N vector. The thickness of lipid bilayer decreased in all systems comprising ILs, while area per lipid increased.


The urokinase plasminogen activator system: a target for anti-cancer therapy.
The urokinase plasminogen activator (uPA) system (uPAS) consists of the uPA, its cognate receptor (uPAR) and two specific inhibitors, the plasminogen activator inhibitor 1 (PAI-1) and 2 (PAI-2). The uPA converts the proenzyme plasminogen in the serine protease plasmin, involved in a number of physiopathological processes requiring basement membrane (BM) and/or extracellular matrix (ECM) remodelling, including tumor progression and metastasis. Data accumulated over the past years have made increasingly clear that the uPAS has a multifunctional task in the neoplastic evolution, affecting tumor angiogenesis, malignant cell proliferation, adhesion and migration, intravasation and growth at the metastatic site. In agreement with their role in cancer progression and metastasis, an increased expression of uPA, uPAR, and PAI-1 has been documented in several malignant tumors, and a positive correlation between the levels of one or more uPAS members and a poor prognosis has been frequently reported. This is particularly evident in breast cancer, for which uPA has been demonstrated to be the most potent independent prognostic factor described to date. The involvement of the uPAS in cancer progression identifies its components as suitable targets for anti-cancer therapy. Several therapeutical approaches aimed at inhibiting the uPA/uPAR functions have been shown to possess anti-tumor effects in xenograft models, including selective inhibitors of uPA activity, antagonist peptides, monoclonal antibodies able to prevent uPA binding to uPAR and gene therapy techniques silencing uPA/uPAR expression. All these strategies, however, although promising, need definitive confirmation in humans as, up to now, only few uPA inhibitors entered clinical trial.


Developmental expression and hormonal regulation of the rat matrix Gla protein (MGP) gene in chondrogenesis and osteogenesis.
Matrix Gla protein (MGP), a vitamin K dependent protein, has recently been identified in many tissues. However, it is accumulated only in bone and cartilage suggesting that the expression of MGP may be related to the development and/or maintenance of the phenotypic properties of these tissues. We systematically evaluated MGP mRNA expression as a function of bone and cartilage development and also as regulated by vitamin D during growth and cellular differentiation. Three experimental models of cartilage and bone development were employed: an in vivo model for endochondral bone formation, as well as in primary cells of normal diploid rat chondrocyte and osteoblast cultures. MGP was expressed at the highest level during cartilage formation and calcification in vivo during endochondral bone formation. In chondrocyte cultures, MGP mRNA was present throughout the culture period but increased only after 3 weeks concomitantly with type I collagen mRNA. In osteoblast cultures, MGP mRNA was expressed during the proliferative period and exhibited increased expression during the period of matrix development. In contrast to osteocalcin (bone Gla protein), this increase was not dependent on mineralization but was related to the extent of differentiation associated with and potentially induced by extracellular matrix formation. During the proliferative period, type I collagen mRNA peaked and thereafter declined, while type I collagen protein steadily accumulated in the extracellular matrix. Constant MGP levels were maintained in the mineralization period of osteoblast differentiation in vitro which is consistent with the constant levels found during the osteogenic period of the in vivo system. MGP mRNA levels in both osteoblasts and chondrocytes in culture were significantly elevated by 1,25-(OH)2D3 (10(-8) M, 48 h) throughout the time course of cellular growth and differentiation. Interestingly, when MGP mRNA transcripts from vitamin D treated and untreated chondrocytes and osteoblasts were analyzed by high resolution Northern blot analysis, we observed two distinct species of MGP mRNA in the vitamin D treated chondrocyte cultures while all other cultures examined exhibited only a single MGP mRNA transcript. Primer extension analysis indicated a single transcription start site in both osteoblasts and chondrocytes with or without vitamin D treatment, suggesting that the lower molecular weight MGP message in vitamin D treated chondrocytes may be related to a modification in post-transcriptional processing. In conclusion, these results show that the selective accumulation of MGP in bone and cartilage tissues in vitro may be related to the development and/or maintenance of a collagenous matrix as reflected by increases in MGP mRNA during these periods.(ABSTRACT TRUNCATED AT 400 WORDS)


Clonogenic assay combined with flow cytometric cell sorting for cell-cycle analysis of human leukemic colony-forming cells.
The purpose of this study is to develop a method combining clonogenic assay and flow cytometric cell sorting based on DNA content for determination of percentage distribution of human malignant clonogenic cells over cell-cycle phases. Flow cytometric cell sorting was carried out for Chinese Hamster Ovary (CHO) and human leukemic bone marrow cells under sterile conditions using a supravital dye for DNA, 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI). The fraction sorted for G0/G1 CHO cells was verified to be almost completely free of G2/M cells and partially cleared of S-phase cells. Cells sorted for G0/G1 fraction were successfully grown as colonies in culture for both CHO and human leukemic marrow cells. The plating efficiency of sorted cells was lower than that of unsorted cells, and the relative contribution of cell damage versus cell selection cannot be determined for this reduction of plating efficiency. The purity of sorted cells must still be improved, and the reason for reduced plating efficiency remains to be defined by further studies. Nonetheless this report represents initial data demonstrating the feasibility of combining two technologies: clonogenic assay and flow cytometric cell sorting based on DNA content. As such, it paves the way for a new approach to studying the cell cycle kinetics of clonogenic cell subpopulations of malignant tissues freshly obtained from patients.


Surgifoam and mitoxantrone in the glioblastoma multiforme postresection cavity: the first step of locoregional chemotherapy through an ad hoc-placed catheter: technical note.
To investigate the safety and feasibility of a novel form of treatment offered by the direct intraoperative application of a Surgifoam-mitoxantrone mix into a glioblastoma multiforme postresection cavity. A technique for the placement of an intracavity catheter connected with a subcutaneous reservoir for further locoregional mitoxantrone administration is also described. Between January and December 2004, 22 consecutive recurrent glioblastoma multiforme patients (14 men, 8 women; age, 56-72 yr; average, 64 yr; median, 65 yr) were enrolled in this study. All patients underwent image-assisted gross total resection of the pathological tissue. A Surgifoam-mitoxantrone mix (1 g Surgifoam powder, 3 ml physiological solution, and 12 mg mitoxantrone in 6 ml) was used to fill the surgical cavity. A ventricular catheter, connected to a Rickham subcutaneous reservoir, was then positioned in the surgical cavity for future mitoxantrone administration. Toxic effects caused by mitoxantrone administration were not observed in any patients during the first postoperative month. On postoperative Days 1, 7, and 30, computed tomographic scans excluded surgical complications. In three patients, residual tumor was disclosed. A mix of Surgifoam and mitoxantrone could be safely applied intraoperatively into the post-glioblastoma multiforme resection cavity without any observable side effects. This technique may benefit both the surgeon and the patient by taking advantage of the drug's hemostatic and cytostatic properties.


Prevalence of Toxoplasma antibodies in pregnant African women in Tanzania.
Three hundred and fifty-seven serum samples from pregnant African women were examined for Toxoplasma gondii antibodies with indirect haemagglutination test in Dar es Salaam. Out of these 222 women were normal, sixty-nine had anaemia and sixty-six were suffering from hypertension. Infection rate in normal pregnant women was 41.9%, in anaemic women 52.5%, and in those suffering from hypertension 66.7%. Highly significant relationship was observed between Toxoplasma infection and anaemia, and hypertension. Infection rate was significantly high in women who had histories of abortion. The results suggested associations of Toxoplasma gondii infection with hypertension and anaemia in African women.


CB2 Receptors and Neuron-Glia Interactions Modulate Neurotoxicity Generated by MAGL Inhibition.
Monoacylglycerol lipase inhibition (MAGL) has emerged as an interesting therapeutic target for neurodegenerative disease treatment due to its ability to modulate the endocannabinoid system and to prevent the production of proinflammatory mediators. To obtain a beneficial response, it is necessary to understand how this inhibition affects the neuron-glia crosstalk and neuron viability. In this study, the effect of MAGL inhibition by KML29 was evaluated in two types of rat cortical primary cultures; mixed cultures, including neuron and glial cells, and neuron-enriched cultures. The risk of neuronal death was estimated by longitudinal survival analysis. The spontaneous neuronal risk of death in culture was higher in the absence of glial cells, a process that was enhanced by KML29 addition. In contrast, neuronal survival was not compromised by MAGL inhibition in the presence of glial cells. Blockade of cannabinoid type 2 (CB2) receptors expressed mainly by microglial cells did not affect the spontaneous neuronal death risk but decreased neuronal survival when KML29 was added. Modulation of cannabinoid type 1 (CB1) receptors did not affect neuronal survival. Our results show that neuron-glia interactions are essential for neuronal survival. CB2 receptors play a key role in these protective interactions when neurons are exposed to toxic conditions.


Profiling gene transcription reveals a deficiency of mitochondrial oxidative phosphorylation in Trypanosoma cruzi-infected murine hearts: implications in chagasic myocarditis development.
In this study, we report the host genetic responses that characterize Trypanosoma cruzi-induced myocarditis in a murine model of infection and disease development. The mRNA species from the myocardium of infected mice were assessed using cDNA microarray technology at immediate early, acute, and chronic stages of infection. The immediate early reaction of the host to T. cruzi infection was marked by up-regulation of transcripts indicative of proinflammatory and interferon-induced immune responses. Following acute infection, overexpression of transcripts for extracellular matrix (ECM) proteins, possibly initiated in response to myocardial injuries by invading and replicating parasites, was suggestive of active reparative and remodeling reactions. Surprisingly, progression to the cardiac disease phase was associated with coordinated down-regulation of a majority (>70%) of the differentially expressed genes. Among the most repressed genes were the troponins, essential for contractile function of the myofibrils, and the genes encoding components of oxidative phosphorylation (OXPHOS) pathways. Reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and biochemical assays confirmed the microarray results and provided evidence for the deficiency of OXPHOS complex IV in the chagasic murine heart. We discuss the apparent role of OXPHOS dysfunction in the cardiac hypertrophic and remodeling processes with the development of chagasic cardiomyopathy (CCM).


Effects of Fisetin Treatment on Cellular Senescence of Various Tissues and Organs of Old Sheep.
Fisetin has been shown to be beneficial for brain injury and age-related brain disease via different mechanisms. The purpose of this study was to determine the presence of senescent cells and the effects of fisetin on cellular senescence in the brain and other vital organs in old sheep, a more translational model. Female sheep 6-7 years old (N = 6) were treated with 100 mg/kg fisetin or vehicle alone on two consecutive days a week for 8 weeks. All vital organs were harvested at the time of sacrifice. Histology, immunofluorescence staining, and RT-Q-PCR were performed on different regions of brain tissues and other organs. Our results indicated that fisetin treatment at the current regimen did not affect the general morphology of the brain. The presence of senescent cells in both the cerebral brain cortex and cerebellum and non-Cornu Ammonis (CA) area of the hippocampus was detected by senescent-associated β-galactosidase (SA-β-Gal) staining and GL13 (lipofuscin) staining. The senescent cells detected were mainly neurons in both gray and white matter of either the cerebral brain cortex, cerebellum, or non-CA area of the hippocampus. Very few senescent cells were detected in the neurons of the CA1-4 area of the hippocampus, as revealed by GL13 staining and GLB1 colocalization with NEUN. Fisetin treatment significantly decreased the number of SA-β-Gal+ cells in brain cortex white matter and GL13+ cells in the non-CA area of the hippocampus, and showed a decreasing trend of SA-β-Gal+ cells in the gray matter of both the cerebral brain cortex and cerebellum. Furthermore, fisetin treatment significantly decreased P16+ and GLB1+ cells in neuronal nuclear protein (NEUN)+ neurons, glial fibrillary acidic protein (GFAP)+ astrocytes, and ionized calcium binding adaptor molecule 1 (IBA1)+ microglia cells in both gray and white matter of cerebral brain cortex. Fisetin treatment significantly decreased GLB1+ cells in microglia cells, astrocytes, and NEUN+ neurons in the non-CA area of the hippocampus. Fisetin treatment significantly decreased plasma S100B. At the mRNA level, fisetin significantly downregulated GLB1 in the liver, showed a decreasing trend in GLB1 in the lung, heart, and spleen tissues, and significantly decreased P21 expression in the liver and lung. Fisetin treatment significantly decreased TREM2 in the lung tissues and showed a trend of downregulation in the liver, spleen, and heart. A significant decrease in NRLP3 in the liver was observed after fisetin treatment. Finally, fisetin treatment significantly downregulated SOD1 in the liver and spleen while upregulating CAT in the spleen. In conclusion, we found that senescent cells were widely present in the cerebral brain cortex and cerebellum and non-CA area of the hippocampus of old sheep. Fisetin treatment significantly decreased senescent neurons, astrocytes, and microglia in both gray and white matter of the cerebral brain cortex and non-CA area of the hippocampus. In addition, fisetin treatment decreased senescent gene expressions and inflammasomes in other organs, such as the lung and the liver. Fisetin treatment represents a promising therapeutic strategy for age-related diseases.


Antigen-Presenting Cells in the Airways: Moderating Asymptomatic Bacterial Carriage.
Bacterial respiratory tract infections (RTIs) are a major global health burden, and the role of antigen-presenting cells (APCs) in mounting an immune response to contain and clear invading pathogens is well-described. However, most encounters between a host and a bacterial pathogen do not result in symptomatic infection, but in asymptomatic carriage instead. The fact that a pathogen will cause infection in one individual, but not in another does not appear to be directly related to bacterial density, but rather depend on qualitative differences in the host response. Understanding the interactions between respiratory pathogens and airway APCs that result in asymptomatic carriage, will provide better insight into the factors that can skew this interaction towards infection. This review will discuss the currently available knowledge on airway APCs in the context of asymptomatic bacterial carriage along the entire respiratory tract. Furthermore, in order to interpret past and futures studies into this topic, we propose a standardized nomenclature of the different stages of carriage and infection, based on the pathogen's position with regard to the epithelium and the amount of inflammation present.


Transcription during the development of bacteriophage phi 29: production of host- and phi 29-specific ribonucleic acid.
The synthesis of ribonucleic acid (RNA) during development of the virulent Bacillus subtilis bacteriophage phi29 has been analyzed. Transcription of host deoxyribonucleic acid (DNA) continues at the preinfection rate throughout the latent period of viral growth. RNA-DNA hybridization was used to show that host messenger RNA synthesis continues late into the phage lytic cycle. Amino acid-labeling experiments show that this RNA is continuously used to produce protein. Ribosomal RNA production is not inhibited by phage infection. Small quantities of phage-specific RNA first appear between min 6 and 9 after infection. This RNA is made exclusively from one of the phi29 DNA strands. At 12 min postinfection, when phage DNA replication commences, large quantities of viral RNA start to be synthesized. This RNA appears to be transcribed from both strands of phi29 DNA. Studies with rifamycin and rifamycin-resistant host strains showed that the production of all phage phi29-specific RNA requires those components of the host RNA polymerase which are sensitive to this antibiotic. Thus, phage phi29 does not stop transcription of host DNA and may produce only one element for regulation of transcription of its own DNA. These findings may reflect the limited amount of genetic information carried by this phage.


High incidence of spontaneous disease in an HLA-DR15 and TCR transgenic multiple sclerosis model.
Multiple sclerosis (MS) is thought to involve CD4 T cell recognition of self myelin, many studies focusing on a pathogenic role for anti-myelin, HLA-DR15-restricted T cells. In experimental allergic encephalomyelitis, it is known which epitopes trigger disease and that disease is associated with determinant spread of T cell reactivity. Characterization of these events in human MS is critical for the development of peptide immunotherapies, but it has been difficult to define the role of determinant spread or define which epitopes might be involved. In this study, we report humanized transgenic mice, strongly expressing HLA-DR15 with an MS-derived TCR; even on a RAG-2 wild-type background, mice spontaneously develop paralysis. Disease, involving demyelination and axonal degeneration, correlates with inter- and intramolecular spread of the T cell response to HLA-DR15-restricted epitopes of myelin basic protein, myelin oligodendrocyte glycoprotein, and alphaB-crystallin. Spread is reproducible and progressive, with two of the epitopes commonly described in responses of HLA-DR15 patients. The fact that this pattern is reiterated as a consequence of CNS tissue damage in mice demonstrates the value of the transgenic model in supplying an in vivo disease context for the human responses. This model, encompassing pathologically relevant, spontaneous disease with the presentation of myelin epitopes in the context of HLA-DR15, should offer new insights and predictions about T cell responses during MS as well as a more stringent test bed for immunotherapies.


Perceived stigma and depression in initially diagnosed pulmonary tuberculosis patients.
To investigate the status of tuberculosis stigma and depression among tuberculosis patients, to examine the relationship between demographics and tuberculosis stigma and depression among tuberculosis patients, and to identify the predictors of depression among tuberculosis patients. After the diagnosis of tuberculosis, patients suffer from disease symptoms and disease-related stigma. Depression is also a major concern in tuberculosis patients. A cross-sectional design was used in this study. A total of 84 subjects, obtained through convenience sampling, enrolled in this study, which was conducted from 1 March 2013-30 December 2014. Data were collected using a structured questionnaire with a demographic component, the Tuberculosis-related Stigma Scale, and the Beck Depression Inventory-II. Data were analysed, using spss Version 20. Independent t tests, Pearson's correlation coefficient tests and analyses of variance were used for analysis of patient demographic characteristics, disease characteristics, and the correlation between stigma and depression. Multiple linear regression was used for determining the predictors of depression. The results showed that not disclosing one's illness to others was associated with tuberculosis stigma and depression. Multiple linear regression analysis indicated that patient-perceived tuberculosis stigma and body mass index accounted for 34% of the variation in depression. These results can serve as a reference for clinical healthcare providers to understand perceived stigma and depression in initially diagnosed tuberculosis patients. An intervention to reduce patient tuberculosis stigma, such as improving community healthcare education or offering mental health outreach, has great potential to lower the level of depression among patients with tuberculosis.


Recent advances in nanomaterials for neural applications: opportunities and challenges.
Nanomedicines are promising for delivering drugs to the central nervous system, though their precision is still being improved. Fortifying nanoparticles with vital molecules can interact with the blood-brain barrier, enabling access to brain tissue. This study summarizes recent advances in nanomedicine to treat neurological complications. The integration of nanotechnology into cell biology aids in the study of brain cells' interactions. Magnetic microhydrogels have exhibited superior neuron activation compared with superparamagnetic iron oxide nanoparticles and hold promise for neuropsychiatric disorders. Nanomaterials have shown notable results, such as tackling neurodegenerative diseases by hindering harmful protein buildup and regulating cellular processes. However, further studies of the safety and effectiveness of nanoparticles in managing neurological diseases and disorders are still required.


The amygdala is not necessary for the familiarity aspect of recognition memory.
Dual-process accounts of item recognition posit two memory processes: slow but detailed recollection, and quick but vague familiarity. It has been proposed, based on prior rodent work, that the amygdala is critical for the familiarity aspect of item recognition. Here, we evaluated this proposal in male rhesus monkeys (Macaca mulatta) with selective bilateral excitotoxic amygdala damage. We used four established visual memory tests designed to assess different aspects of familiarity, all administered on touchscreen computers. Specifically, we assessed monkeys' tendencies to make low-latency false alarms, to make false alarms to recently seen lures, to produce curvilinear ROC curves, and to discriminate stimuli based on repetition across days. Three of the four tests showed no familiarity impairment and the fourth was explained by a deficit in reward processing. Consistent with this, amygdala damage did produce an anticipated deficit in reward processing in a three-arm-bandit gambling task, verifying the effectiveness of the lesions. Together, these results contradict prior rodent work and suggest that the amygdala is not critical for the familiarity aspect of item recognition.


Tuberculosis Mortality by Occupation in South Africa, 2011⁻2015.
Work-related tuberculosis (TB) remains a public health concern in low- and middle-income countries. The use of vital registration data for monitoring TB deaths by occupation has been unexplored in South Africa. Using underlying cause of death and occupation data for 2011 to 2015 from Statistics South Africa, age-standardised mortality rates (ASMRs) were calculated for all persons of working age (15 to 64 years) by the direct method using the World Health Organization (WHO) standard population. Multivariate logistic regression analysis was performed to calculate mortality odds ratios (MORs) for occupation groups, adjusting for age, sex, year of death, province of death, and smoking status. Of the 221,058 deaths recorded with occupation data, 13% were due to TB. ASMR for TB mortality decreased from 165.9 to 88.8 per 100,000 population from 2011 to 2015. An increased risk of death by TB was observed among elementary occupations: agricultural labourers (MORadj = 3.58, 95% Confidence Interval (CI) 2.96⁻4.32), cleaners (MORadj = 3.44, 95% CI 2.91⁻4.09), and refuse workers (MORadj = 3.41, 95% CI 2.88⁻4.03); among workers exposed to silica dust (MORadj = 3.37, 95% CI 2.83⁻4.02); and among skilled agricultural workers (MORadj = 3.31, 95% CI 2.65⁻4.19). High-risk TB occupations can be identified from mortality data. Therefore, TB prevention and treatment policies should be prioritised in these occupations.


Clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum in horses with experimentally induced osteoarthritis.
To assess the clinical, biochemical, and histologic effects of intra-articular administration of autologous conditioned serum (ACS) in the treatment of experimentally induced osteoarthritis in horses. 16 horses. Osteoarthritis was induced arthroscopically in 1 middle carpal joint of all horses. In 8 placebo- and 8 ACS-treated horses, 6 mL of PBS solution or 6 mL of ACS was injected into the osteoarthritis-affected joint on days 14, 21, 28, and 35, respectively; PBS solution was administered in the other sham-operated joints. Evaluations included clinical assessment of lameness and synovial fluid analysis (performed biweekly); gross pathologic and histologic examinations of cartilage and synovial membrane samples were performed at necropsy. No adverse treatment-related events were detected. Horses that were treated with ACS had significant clinical improvement in lameness, unlike the placebo-treated horses. Among the osteoarthritis-affected joints, ACS treatment significantly decreased synovial membrane hyperplasia, compared with placebo-treated joints; although not significant, the ACS-treated joints also appeared to have less gross cartilage fibrillation and synovial membrane hemorrhage. The synovial fluid concentration of interleukin-1 receptor antagonist (assessed by use of mouse anti-interleukin-1 receptor antagonist antibody) was increased following treatment with ACS. Results of this controlled study indicated that there was significant clinical and histologic improvement in osteoarthritis-affected joints of horses following treatment with ACS, compared with placebo treatment. On the basis of these findings, further controlled clinical trials to assess this treatment are warranted, and investigation of the mechanisms of action of ACS should be pursued concurrently.


Multiple target sites for designing candidate drugs.
Rational drug discovery strategy requires a design of small molecules as candidate drugs which can specifically inhibit a target protein or any other macromolecule and effectively interfere in a defined physiological process. One of the important bacterial protein targets aimed toward developing new antibiotics is peptidyl-tRNA hydrolase (Pth). The discovery that cytarabine, a known anticancer drug, binds to Pth from Acinetobacter baumannii in a cleft located away from the catalytic site of this enzyme, published in Biochemical Journal, opens up interesting new avenues for drug design. An approach involving crystallographic identification of multiple ligand-binding sites on a target protein surface could enable iterative optimization of multiple high-affinity ligands, which may synergistically interfere in the target function with enhanced effect.


Photoperiodic induction without light-mediated circadian entrainment in a High Arctic resident bird.
Organisms use changes in photoperiod to anticipate and exploit favourable conditions in a seasonal environment. While species living at temperate latitudes receive day length information as a year-round input, species living in the Arctic may spend as much as two-thirds of the year without experiencing dawn or dusk. This suggests that specialised mechanisms may be required to maintain seasonal synchrony in polar regions. Svalbard ptarmigan (Lagopus muta hyperborea) are resident at 74-81°N latitude. They spend winter in constant darkness (DD) and summer in constant light (LL); extreme photoperiodic conditions under which they do not display overt circadian rhythms. Here, we explored how Arctic adaptation in circadian biology affects photoperiodic time measurement in captive Svalbard ptarmigan. For this purpose, DD-adapted birds, showing no circadian behaviour, either remained in prolonged DD, were transferred into a simulated natural photoperiod (SNP) or were transferred directly into LL. Birds transferred from DD to LL exhibited a strong photoperiodic response in terms of activation of the hypothalamic thyrotropin-mediated photoperiodic response pathway. This was assayed through expression of the Eya3, Tshβ and deiodinase genes, as well as gonadal development. While transfer to SNP established synchronous diurnal activity patterns, activity in birds transferred from DD to LL showed no evidence of circadian rhythmicity. These data show that the Svalbard ptarmigan does not require circadian entrainment to develop a photoperiodic response involving conserved molecular elements found in temperate species. Further studies are required to define how exactly Arctic adaptation modifies seasonal timer mechanisms.


Species Diversity, Phylogeny and Large Scale Biogeographic Patterns of the Genus Padina (Phaeophyceae, Dictyotales).
The brown algal genus Padina (Dictyotales, Phaeophyceae) is distributed worldwide in tropical and temperate seas. Global species diversity and distribution ranges, however, remain largely unknown. Species-level diversity was reassessed using DNA-based, algorithmic species delineation techniques based on cox3 and rbcL sequence data from 221 specimens collected worldwide. This resulted in estimates ranging from 39 to 61 putative species (ESUs), depending on the technique as well as the locus. We discuss the merits, potential pitfalls, and evolutionary and biogeographic significance of algorithmic species delineation. We unveil patterns whereby ESUs are in all but one case restricted to either the Atlantic or Indo-Pacific Ocean. Within ocean basins we find evidence for the vast majority of ESUs to be confined to a single marine realm. Exceptions, whereby ESUs span up to three realms, are located in the Indo-Pacific Ocean. Patterns of range-restricted species likely arise by repeated founder events and subsequent peripatric speciation, hypothesized to dominate speciation mechanisms for coastal marine organisms in the Indo-Pacific. Using a three-gene (cox3, psaA and rbcL), relaxed molecular clock phylogenetic analysis we estimated divergence times, providing a historical framework to interpret biogeographic patterns.


Pharmacokinetically based mapping device for chemical space navigation.
ChemGPS, the chemical global positioning system, is a tool that combines rules (equivalent to dimensions) and objects (chemical structures) to provide a consistent chemical space map (Oprea, T. I.; Gottfries, J. J. Comb. Chem. 2001, 3, 157-166.). Rules included, initially, general properties such as size, lipophilicity, and hydrogen bond capacity, while objects include "satellites", intentionally placed outside the druglike space, as well as "core" objects, mostly orally available drugs. ChemGPS molecules (objects) were used in conjunction with the VolSurf (http://www.moldiscovery.com) descriptors (rules), which are relevant for ADME (absorption, distribution, metabolism, and excretion) properties. The combination of ChemGPS and VolSurf, GPSVS, was investigated with respect to the biopharmaceutics classification system, which is recommended by the Food and Drug Administration (FDA) (http://www.fda.gov/cder/OPS/BCS_guidance.htm), in particular with respect to permeability and solubility. The first GPSVS principal component correlates, with no further training, to passive transcellular permeability, as illustrated for the Caco-2, ghost erythrocyte, and blood-brain barrier datasets, respectively. The second GPSVS principal component correlates, without prior training, to solubility, as shown for the octanol-water partition and intrinsic solubility datasets, respectively. Although derived from principal component analysis, the two property axes rotate and form an angle of approximately 43 degrees, thus being no longer orthogonal. GPSVS can be used to map the chemical space with respect to permeability and solubility, as recommended by FDA's biopharmaceutics classification system.


A novel intracellular protein delivery platform based on single-protein nanocapsules.
An average cell contains thousands of proteins that participate in normal cellular functions, and most diseases are somehow related to the malfunctioning of one or more of these proteins. Protein therapy, which delivers proteins into the cell to replace the dysfunctional protein, is considered the most direct and safe approach for treating disease. However, the effectiveness of this method has been limited by its low delivery efficiency and poor stability against proteases in the cell, which digest the protein. Here, we show a novel delivery platform based on nanocapsules consisting of a protein core and a thin permeable polymeric shell that can be engineered to either degrade or remain stable at different pHs. Non-degradable capsules show long-term stability, whereas the degradable ones break down their shells, enabling the core protein to be active once inside the cells. Multiple proteins can be delivered to cells with high efficiency while maintaining low toxicity, suggesting potential applications in imaging, therapy and cosmetics fields.


Heart rate variability of preterm neonates quantified by energy entropy.
Identifying variables predictive of neurobehavioural sequelae is a key objective in the study of high-risk neonates. Examination of heart rate variability (HRV) characteristics may be a finer discriminator of the neonate's response to physiologic stressors than the mean heart rate. The energy entropy of the heart beat tachogram, computed in four different domains, was used to quantify the HRV in 13 preterm neonates. The entropies of energies were computed from 1024 interbeat time intervals obtained once per week from 26 to 35 weeks postconceptional age (PCA). The energy entropy computed in three of the domains, like the standard deviation of intervals, distinguished between the 10 neonates that were measured at 35 weeks PCA with 100% specificity and 67% sensitivity, but did not distinguish between healthy and unhealthy neonates at earlier ages. The findings suggest that energy entropy may be a discerning measure of physiologic stress in the preterm infant, although future research is needed to refine the test and determine statistical significance.


Application of some polymers in the physiocochemical design of tablet formulation.
Polyethylene glycol 4000 did not apprecialbly affect the rate of drug release from tablets. The use of sodium alginate the a tablet binder showed variable effects on the rate of drug release from the tablet i.e. The incorporation of only 2.5% of sodium alginate caused enhancement in the release rate of the drug based on its action as disintegration inducer. On the other hand, the incorporation of 20, 35 and 50% of sodium alginate showed a pronounced retardation in the rate of drug release from the tablets. This retardation was more pronounced in acid than in alkaline media. Accordingly, sodium alginate could be favourably suggested as tablet binder when retarded gastric absorption is the therapeutic aim required. Carbopol 940 when used as a tablet binder in a concentration of 2.5%, showed a marked enhancement of the release rate of the drug. On the contrary, it exerted a remarkable retardation in the rate of drug release from the tablets when incorporated in the relatively large proportions of 20, 35 and 50%. This retardation was more pronounced in alkaline than in acid media. Accordingly, Carbopol 940 would be favourably suggested in the physiocohemical design of prolonged-release tablet formulations.


Interdiagnostician Reliability of the DSM-5 Section II and Section III Alternative Model Criteria for Borderline Personality Disorder.
This study examined the interdiagnostician reliability and potential gender bias of the DSM-IV/DSM-5 Section II and DSM-5 Alternative Model definitions of borderline personality disorder. A national sample of 123 mental health professionals provided diagnostic judgments on 12 case vignettes selected to represent a range of personality pathology. Two versions of each case were included, one identified as male and the other as female, but which were otherwise identical. Analyses examined the intraclass correlation between clinicians and also examined rates of diagnostic assignments as a function of case gender. Reliability of diagnosis of borderline personality did not differ across the two diagnostic approaches, and concordance of diagnoses across the two systems was significant. The dimensional components of the DSM-5 Alternative Model demonstrated significantly more diagnostic reliability than the DSM-IV categorical diagnoses. The DSM-5 Alternative Model conceptualization of borderline personality can be diagnosed with comparable or greater reliability than the extant DSM-IV definition.


Atomic force microscopy study of the antibacterial effects of chitosans on Escherichia coli and Staphylococcus aureus.
Chitosan has been reported to be a non-toxic, biodegradable antibacterial agent. The aim of this work was to elucidate the relationship between the molecular weight of chitosan and its antimicrobial activity upon two model microorganisms, one Gram-positive (Staphylococcus aureus) and one Gram-negative (Escherichia coli). Atomic force microscopy (AFM) imaging was used to obtain high-resolution images of the effect of chitosans on the bacterial morphology. The AFM measurements were correlated with viable cell numbers, which show that the two species reacted differently to the high- and low-molecular-weight chitosan derivatives. The images obtained revealed not only the antibacterial effects, but also the response strategies used by the bacteria; cell wall collapse and morphological changes reflected cell death, whereas clustering of bacteria appeared to be associated with cell survival. In addition, nanoindentation experiments with the AFM revealed mechanical changes in the bacterial cell wall induced by the treatment. The nanoindentation results suggested that despite little modification observed in the Gram-positive bacteria in morphological studies, cell wall damage had indeed occurred, since cell wall stiffness was reduced after chitooligosaccharide treatment.


Small drug sample fabrication of controlled release polymers using the microextrusion method.
Ethylene vinylacetate polymer (EVA) has been used for many years to fabricate controlled-release polymeric implant devices with which drugs of high or low molecular weight compounds could be delivered with zero-order kinetics. However, because the known fabrication methods such as solvent evaporation, casting and possible shrinkage are not sufficiently controllable we have now developed the microextrusion method with which even small amount of clinically important and expensive drugs can be incorporated into EVA with high reproducibility. We show here that devices produced by the microextrusion method allows for a controlled delivery of several neurotoxic and neurotherapeutic compounds such as alpha-methyl-p-tyrosine, diazepam, quinolinic acid, and phencyclidine. Each substance is slowly released from the polymer, as evidenced by spectrophotometric data, for up to 120 days at daily rates varying from 18.4 microg of phencyclidine to 97.6 microg/day of diazepam. Thus, microextrusion is a valuable method for fabricating controlled-release polymers in which small amounts of scarce drugs can be incorporated. Another advantage of the current procedure is that polymers can be fabricated with very little amount of solvent.


Mutational spectrum and genotype-phenotype correlations in mevalonate kinase deficiency.
Mevalonate kinase deficiency (MKD) is an autosomal recessive autoinflammatory disorder caused by mutations in the MVK gene resulting in deficient activity of mevalonate kinase (MK). Depending on the clinical severity, MKD may present as hyper-IgD and periodic fever syndrome (HIDS) or the more severe mevalonic aciduria (MA). We analyzed the MVK gene in 57 patients with MKD and found 39 different mutations including 15 novel mutations, expanding the total mutational spectrum of MKD to 63 mutations. To get more insight into the genotype-phenotype correlation in MKD, we studied the effect of selected missense mutations on MK protein stability and activity in various patient fibroblast cell lines. All MKD cell lines showed markedly decreased MK activities that correlated well with the clinical severity and, for most of the cell lines, with the amount of MK protein. When fibroblasts of MKD patients were cultured under conditions known to promote a more controlled protein folding, all cell lines of patients with the HIDS phenotype and few cell lines of patients with the MA phenotype showed an increase in the residual MK activity. This increase in enzyme activity correlates well with an increase in the MK protein levels in these cell lines, indicating that most of the mutations in MKD affect stability and/or folding of the MK protein rather than affecting the catalytic properties of the enzyme. The finding that the residual activity in MKD can be manipulated by environmental conditions may offer therapeutic options to alleviate or prevent the clinical symptoms associated with MKD.


Intraoperative motor-evoked potential monitoring during coil embolization for anterior choroidal artery aneurysms.
Intraoperative motor-evoked potential (MEP) monitoring is widely used in the neck clipping of cerebral aneurysms. Little is known regarding the usefulness of intraoperative MEP monitoring in endovascular aneurysm surgery. The purpose of this study was to validate the feasibility of intraoperative MEP monitoring during the coil embolization of anterior choroidal artery (AChA) aneurysms. Clinical and angiographic data of consecutive patients who underwent coil embolization for unruptured AChA aneurysms with or without intraoperative MEP monitoring between January 2014 and December 2018 at our institute were abstracted and analyzed retrospectively. Twenty-three unruptured AChA aneurysms were treated. Eleven patients received MEP monitoring, and three of them experienced intraoperative reduction or disappearance of the MEP wave. Even during MEP changes, AChA filling showed no change in any of the three cases. Although one case with MEP monitoring encountered the disappearance of AChA filling, there was no change in MEP. This might be due to retrograde filling of the AChA from the anastomosis with the lateral posterior choroidal artery. AChA blood flow detected by angiography did not always reflect MEP status. When comparing the presence or absence of MEP monitoring, the volume embolization ratio of coiled aneurysms was significantly better in the MEP group. Intraoperative MEP monitoring during endovascular coiling for AChA aneurysms may be feasible. AChA blood flow detected by angiography does not always reflect MEP status.


Diversity and fluctuation of spine morphology in CA1 pyramidal neurons after transient global ischemia.
Dendritic spines form postsynaptic components of excitatory synapses in CA1 pyramidal neurons and play a key role in excitatory signal transmission. Transient global ischemia is thought to induce excitotoxicity that triggers delayed neuronal death in the CA1 region. However, the mechanism underlying structural changes of excitatory synapses after ischemia is not completely understood. Here, we demonstrate how dendritic spines change in their density and structure at an acute stage after transient global ischemia. Intracellular staining in vivo showed that the total spine density in basal, proximal, and distal apical dendrites increased at 12 hr and 24 hr after ischemia, but returned to control levels at 48 hr after ischemia. Consistent increase of spine density mainly appeared in non-late depolarizing postsynaptic potential neurons, although late depolarizing postsynaptic potential neurons also showed slight increases in spine density in these dendrites at the same intervals after ischemia. Golgi staining showed increased spine density occurred in less swollen dendrites but decreased spine density appeared in severely swollen dendrites at 12 and 24 hr after ischemia. In addition, the density and percentage of stubby spines reduced at 12 hr and 48 hr, whereas the density of thin spines increased at 12 hr after ischemia. The density and percentage of filopodia increased nearly fivefold at 24 hr after ischemia. Moreover, the density of mushroom spines doubled and its percentage increased by 150% at 48 hr after ischemia. These morphological changes of spines may be related to neuronal injury in CA1 pyramidal neurons after ischemia.


[Akatinol memantine in the therapy of cognitive disorders in Parkinson's disease].
Thirty-eight patients, 20 male, 18 female, mean age 61.8+/-7.7 years, illness duration 8.4+/-1.3 years, have been studied. The degree of severity of Parkinson's disease (PD) according to the Hoehn and Yahr scale was 2.6+/-0.4. Mild cognitive impairment was observed in all cases. Akatinol memantine was prescribed to patients, who have been receiving levodopa, in the following dosages: the first week--5mg daily, the second week--10 mg daily and from the third week--20 mg in fractional dosages. Treatment duration was 6 months. The patient's state before and after treatment was assessed clinically and with several scales measuring cognitive and motor functions. It was shown that akatinol memantine not only reduced disorders typical for PD (hypokinesia, rigidity, tremor) and increased the daily activity of patients but significantly improved the indices of logic and visual memory, speech functions and thinking.


Recovery expectations and long-term prognosis of patients with coronary heart disease.
Expectations of patients regarding their prospects for recovery have been shown to predict subsequent physical and social functioning. Evidence regarding the impact of expectations on clinical outcomes is limited. At the inpatient service of a tertiary care hospital, we evaluated beliefs of patients undergoing coronary angiography about their prognosis as predictors of long-term survival and 1-year functional status. Baseline assessments, including a measure of expectations for recovery, were obtained during hospitalization with mortality follow-up for approximately 15 years. Patients with significant obstructive coronary artery disease were interviewed while in the hospital and enrolled in follow-up. Functional status was assessed at baseline and 1 year later with questionnaires reflecting physical capabilities. Analyses controlled for age, sex, disease severity, comorbidities, treatments, demographics, depressive symptoms, social support, and functional status. There were 1637 total deaths, 885 from cardiovascular causes, in the 2818 patients in these analyses. The outcomes were total mortality, cardiovascular mortality, and 1-year functional status. Expectations were positively associated with survival after controlling for background and clinical disease indicators. For a difference equivalent to an interquartile range of expectations, the hazard ratio (HR) for total mortality was 0.76 (95% confidence interval [CI], 0.71-0.82) and 0.76 (95% CI, 0.69-0.83) for cardiovascular mortality. The HRs were 0.83 (95% CI, 0.76-0.91) and 0.79 (95% CI, 0.70-0.89) with further adjustments for demographic and psychosocial covariates. Similar associations (P < .001) were observed for functional status. Recovery expectations at baseline were positively associated with long-term survival and functioning in patients with coronary artery disease.


Treatment of hypertension in peripheral arterial disease.
Peripheral arterial disease (PAD) causes considerable morbidity and mortality. Hypertension is a risk factor for PAD. Treatment for hypertension must be compatible with the symptoms of PAD. Controversy regarding the effects of beta-adrenoreceptor blockade for hypertension in patients with PAD has led many physicians to stop prescribing beta-adrenoreceptor blockers. Little is known about the effects of other classes of anti-hypertensive drugs in the presence of PAD. This is the second update of a Cochrane review first published in 2003. To determine the effects of anti-hypertensive drugs in patients with both raised blood pressure and symptomatic PAD in terms of the rate of cardiovascular events and death, symptoms of claudication and critical leg ischaemia, and progression of atherosclerotic PAD as measured by ankle brachial index (ABI) changes and the need for revascularisation (reconstructive surgery or angioplasty) or amputation. For this update the Cochrane Peripheral Vascular Diseases Group Trials Search Co-ordinator searched the Cochrane Peripheral Vascular Diseases Group Specialised Register (last searched March 2013) and CENTRAL (2013, Issue 2). Randomised controlled trials (RCTs) of at least one anti-hypertensive treatment against placebo or two anti-hypertensive medications against each other, with interventions lasting at least one month. Trials had to include patients with symptomatic PAD. Data were extracted by one author (DAL) and checked by the other (GYHL). Potentially eligible studies were excluded when the results presentation prevented adequate extraction of data and enquiries to authors did not yield raw data. Eight RCTs were included with a total of 3610 PAD patients. Four studies compared a recognised class of anti-hypertensive treatment with placebo and four studies compared two anti-hypertensive treatments with each other. Studies were not pooled due to the variation of the comparisons and the outcomes presented. Overall the quality of the available evidence was unclear, primarily as a result of a lack of detail in the study reports on the randomisation and blinding procedures and incomplete outcome data. Two studies compared angiotensin converting enzyme (ACE) inhibitors against placebo. In one study there was a significant reduction in the number of cardiovascular events in patients receiving ramipril (odds ratio (OR) 0.72, 95% confidence interval (CI) 0.58 to 0.91; n = 1725). In the second trial using perindopril (n = 52) there was a marginal increase in claudication distance but no change in ABI and a reduction in maximum walking distance. A trial comparing the calcium antagonist verapamil versus placebo in patients undergoing angioplasty (n = 96) suggested that verapamil reduced restenosis (per cent diameter stenosis (± SD) 48.0% ± 11.5 versus 69.6% ± 12.2; P < 0.01), although this was not reflected in the maintenance of a high ABI (0.76 ± 0.10 versus 0.72 ± 0.08 for verapamil versus placebo). Another study (n = 80) demonstrated no significant difference in arterial intima-media thickness (IMT) in men receiving the thiazide diuretic hydrochlorothiazide (HCTZ) compared to those receiving the alpha-adrenoreceptor blocker doxazosin (-0.12 ± 0.14 mm and -0.08 ± 0.13 mm, respectively; P = 0.66). A study (n = 36) comparing telmisartan to placebo found a significant improvement in maximum walking distance at 12 months with telmisartan (median (interquartile range (IQR)) 191 m (157 to 226) versus 103 m (76 to 164); P < 0.001) but no differences in ABI (median (IQR) 0.60 (0.60 to 0.77) versus 0.52 (0.48 to 0.67)) or arterial IMT (median (IQR) 0.08 cm (0.07 to 0.09) versus 0.09 cm (0.08 to 0.10)). Two studies compared the beta-adrenoreceptor blocker nebivolol with either the thiazide diuretic HCTZ or with metoprolol. Both studies found no significant differences in intermittent or absolute claudication distance, ABI, or all-cause mortality between the anti-hypertensives. A subgroup analysis of PAD patients (n = 2699) in a study which compared a calcium antagonist-based strategy (verapamil slow release (SR) ± trandolapril) to a beta-adrenoreceptor blocker-based strategy (atenolol ± hydrochlorothiazide) found no significant differences in the composite endpoints of death, non-fatal myocardial infarction or non-fatal stroke with or without revascularisation (OR 0.90, 95% CI 0.76 to 1.07 and OR 0.96, 95% CI 0.82 to 1.13, respectively). Evidence on the use of various anti-hypertensive drugs in people with PAD is poor so that it is unknown whether significant benefits or risks accrue. However, lack of data specifically examining outcomes in PAD patients should not detract from the overwhelming evidence on the benefit of treating hypertension and lowering blood pressure.


Transport and anatomic distribution of a high molecular weight fraction of bovine pituitary intraglandular colloid of intermediate lobe origin.
Bovine pituitary intraglandular colloid (IGC) of intermediate lobe (IL) origin, was separated into seven primary fractions (F1-F7) on a 2.6 X 70 cm column of dextran gel G-100 superfine (Pharmacia). The molecular weight of F1-F3 was much greater than 1000 000; F4, 63 000; F5, 34 000 and F6-F7 less than 4000. The first fraction (F1) was used in this study and was labelled with (125I)Na via a modification of the chloramine-T method. Male Wistar rats were divided into 5 experimental and 5 control groups. The animals were fed Purina Lab Chow and Lugol's iodine was mixed with the drinking water to minimize any uptake by the thyroid gland of 125I liberated by metabolism. Under anesthesia, the animals in the experimental group were administered via the right internal jugular vein, 0.15 ml of (125I)F1 and those in the control were administered with 0.15 ml of free (125I)NA. Animals were killed with sodium pentobarbital at 2, 4, 6, 12 and 24 h periods. There are 5 replications for each of the 5 treatments (time periods) in every experiment including the control. Twenty-one tissues were retrieved and the kidney, spleen and thyroid were the only tissues that had retained the (125I)F1 at the 24 h recording period. Autoradiographic labelling was detected in hematoxylin-eosin stained sections of these tissues. Although it is difficult to make an exact determination as to whether the labelled fraction recovered is the same as the one injected, the kidney, spleen and thyroid continued to be significant tissues after several repeated experiments. The autoradiographic picture was the same each time the experiment was repeated. These results tend to indicate that IGC of IL origin is involved in the transport of IL materials.


Pancastatin A and B Have Selective Cytotoxicity on Glucose-Deprived PANC-1 Human Pancreatic Cancer Cells.
Glucose deprivation and hypoxia frequently occur in solid tumor cells, including pancreatic cancer cells. Glucose deprivation activates the unfolded protein response (UPR) and causes the upregulation of glucose-regulated protein 78 (GRP78). Induction of GRP78 has been shown to protect cancer cells. Therefore, shutting down of GRP78 expression may be a novel strategy in anticancer drug development. Based on this understanding, a screening system established for anticancer agents that exhibit selective cytotoxicity on pancreatic cancer cells under glucose-deprived conditions. To test this hypothesis, the new compounds isolated, pancastatin A (PST-A) and B (PSTB), from Ponciri Fructus. PST-A and B were identified as glabretal triterpenoid moieties by electrospray ionization mass spectrometry and nuclear magnetic resonance spectroscopic methods. PST-A and B suppressed the accumulation of the UPR hallmark gene, GRP78, during glucose deprivation. Furthermore, PST-A and B showed selective cytotoxicity on PANC-1 pancreatic cancer cells under glucose deprivation. Interestingly, PST-A and B had no effect on these cells under normal growth conditions. Our results suggest that PST-A and B act as novel therapeutic agents to induce selective cell death in glucose-deprived pancreatic cancer cells.


ZNRF2 is released from membranes by growth factors and, together with ZNRF1, regulates the Na+/K+ATPase.
Here, we describe a phosphorylation-based reverse myristoyl switch for mammalian ZNRF2, and show that this E3 ubiquitin ligase and its sister protein ZNRF1 regulate the Na(+)/K(+) pump (Na(+)/K(+)ATPase). N-myristoylation localizes ZNRF1 and ZNRF2 to intracellular membranes and enhances their activity. However, when ZNRF2 is phosphorylated in response to agonists including insulin and growth factors, it binds to 14-3-3 and is released into the cytosol. On membranes, ZNRF1 and ZNRF2 interact with the Na(+)/K(+)ATPase α1 subunit via their UBZ domains, while their RING domains interact with E2 proteins, predominantly Ubc13 that, together with Uev1a, mediates formation of Lys63-ubiquitin linkages. ZNRF1 and ZNRF2 can ubiquitylate the cytoplasmic loop encompassing the nucleotide-binding and phosphorylation regions of the Na(+)/K(+)ATPase α1 subunit. Ouabain, a Na(+)/K(+)ATPase inhibitor and therapeutic cardiac glycoside, decreases ZNRF1 protein levels, whereas knockdown of ZNRF2 inhibits the ouabain-induced decrease of cell surface and total Na(+)/K(+)ATPase α1 levels. Thus, ZNRF1 and ZNRF2 are new players in regulation of the ubiquitous Na(+)/K(+)ATPase that is tuned to changing demands in many physiological contexts.


Effects of cyclocryosurgery on the clinically normal canine eye.
Four cyclocryosurgical procedures were compared for effects on normal canine eyes. All procedures produced a significant (P less than 0.05) decrease in intraocular pressure, a marked uveitis, ciliary epithelial cell loss, edema and engorgement of the ciliary processes, iris necrosis, protein effusion, chemosis, and mild discomfort, as indicated by mild blepharospasm and slight epiphora. The histopathologic changes were evaluated at 5 minutes, 1 week, 1 month, and 6 months after surgery. The ciliary epithelium was normal in appearance and intraocular pressure had returned to the normal range at 6 months after surgery. The adverse reactions to the cyclocryosurgical procedures included retinal detachments, chemosis, conjunctivitis, transient increased intraocular pressure, uveitis, iris depigmentation, and corneal granulation tissue.


Changes in pelvic floor electrical activity and vulvar pain after botulinum toxin treatment of vestibulodynia: are clinical and electrophysiological outcomes related?
Vestibulodynia is a gynecological condition with different treatment options available, including botulinum neurotoxin type A (BoNT/A) injections into the vulvar vestibule. Unlike other treatments, no studies have assessed changes in the myoelectrical activity of the pelvic floor muscles (PFM) after BoNT/A treatment. The aim of this study was thus to evaluate these changes and to correlate them with changes in vulvar pain sensitivity. To do this, 35 patients with vestibulodynia were recruited, the myoelectrical activity of their left and right PFM was recorded with surface electromyography (sEMG), and their vulvar pain sensitivity was monitored according to Visual Analogue Scale (VAS) and an algometer, both before and after BoNT/A treatment. According to our results, patients' signals during PFM relaxation showed a significantly higher power than those of healthy women at baseline, as shown by their root mean square values (RMS), but became similar at follow-up. Patients' mean vulvar pain VAS scores significantly decreased after treatment. Furthermore, baseline-to-follow-up differences of RMS at PFM rest vs. mean VAS were significantly correlated (CC=0.48, p<0.01) so that higher reductions in the PFM activity power were associated with higher decreases in vulvar pain.Clinical Relevance- Altered PFM electrophysiological condition of patients with vestibulodynia becomes similar to healthy women's after BoNT/A treatment. This study also points to a relationship between the evolution of clinical and PFM electrophysiological conditions.


Enzymatic modification for improving nutritional qualities and acceptability of proteins extracted from photosynthetic microorganisms Spirulina maxima and Rhodopseudomonas capsulatus.
The present study attempts to improve the proteins from a blue-green alga Spirulina maxima and a non-sulfur purple bacterium Rhodopseudomonas capsulatus through their peptic hydrolysis followed by plastein synthesis with papain. The former enzymatic process was effective in removing some photosynthetic pigments and flavors originating in the raw materials. The latter process was successful in incorporating limited amounts of methionine, lysine, and tryptophan, and thus to synthesize plasteins whose essential amino acid patterns resemble the FAO/WHO suggested pattern (1973). These plasteins had no colors and no flavors.


Isolation and characterization of lactic acid bacteria from jiang-sun (fermented bamboo shoots), a traditional fermented food in Taiwan.
In this study, lactic acid bacteria (LAB) were isolated, characterized and identified from jiang-sun (fermented bamboo shoots; a traditional fermented food in Taiwan). Samples were collected at seven time intervals from a fixed fermenting bucket during the fermentation process of jiang-sun and its initial ingredients, dochi (fermented soybeans) and bamboo shoots. A total of 57 LAB cultures were isolated; 42 cultures were isolated from jiang-sun samples and 15 cultures were isolated from dochi and bamboo shoots. These isolates were characterized phenotypically and then divided into three groups (A-C) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Alteration of microbial populations during the fermentation process was observed. While various LAB were found in the dochi and bamboo shoots, it was mostly replaced by Lactobacillus plantarum after 1 day of fermentation. Furthermore, the antibacterial activities of the isolates were determined, and one Enterococcus faecium strain showed inhibitory activity against all the indicator strains. Results suggest that L. plantarum is the main LAB present during the fermentation of jiang-sun. To our knowledge, this is the first report describing the distribution and varieties of LAB that exist in the jiang-sun fermentation process.


Determinants of mortality in pediatric patients with greater than 70% full-thickness total body surface area thermal injury treated by early total excision and grafting.
Recent advancements in burn care have improved the survival rates of victims with severe burn injuries. The total mortality rate in a series of 1,057 pediatric patients admitted to Shriners Burns Institute Galveston Unit between 1982 and 1986 was 2.7%. The burn size resulting in a 50% death rate was 95% of the total body surface area (TBSA). In this study 19 survivors and 13 nonsurvivors with greater than 70% full-thickness TBSA burn injuries were compared. All survivors were adequately resuscitated upon arrival 11% sustained an inhalation injury. Forty-six per cent of the nonsurvivors sustained an inhalation injury; 31% were not initially adequately resuscitated. The presence of preadmission shock and inhalation injury were early determinants of mortality with secondary renal, pulmonary, or cardiovascular collapse being the later predictors of mortality in these massively burned pediatric patients.


Regulation of bacterial growth.
Is the control of bacterial metabolism so complex? The answer can be found in a simple experiment. Two cultures of bacteria are grown in different mediums. One contains as the carbon and nitrogen sources a mixture of amino acids, while the other contains only glucose and ammonia, so that the cells must synthesize all of the amino acids. The results show that insofar as the cells in both cultures grow at comparable rates, they will have the same composition in terms of DNA, RNA, and protein (30). To explain this phenomena I have argued that through the control mechanisms responsible for the distribution of substrates in intermediary metabolism, the substrates of protein synthesis are produced at concentrations and rates commensurate with the ability of the environment to support growth. The provision of these substrates relative to the ability of the protein forming system to utilize them regulates the synthesis of ribosomal and transfer RNA, which, after adjustment for various modulating influences, such as nonfunctioning ribosomes or ribosomal RNA turnover, brings the number of functioning ribosomes to a point in keeping with the provision of external nutrients. The synthesis of messenger (or total) RNA, ribosomal proteins, and DNA, and the process of cell division, for example, are subject to their own controls, but through the burden they each place on intermediary metabolism, they provide a means for partitioning the cell's metabolic resources. It might be noted that this view may not be very far from the idea once held that the rate at which each of the transfer RNA's was changed by amino acids regulate the synthesis of bacterial RNA, but growth regulation is clearly more complicated than implied by that model (76).


The ontogeny of the homing pigeon navigational map: evidence for a sensitive learning period.
Homing pigeons can learn a navigational map by relying on the heterogeneous distribution of atmospheric odours in the environment. To test whether there might be a sensitive period for learning an olfactory-based navigational map, we maintained a group of young pigeons in an aviary screened from the winds until the age of three to four months post-fledging. Subsequently, the screens were removed and the pigeons were exposed to the winds and the environmental odours they carry for three months. One control group of pigeons was held in a similar aviary but exposed to the winds immediately upon Hedging, while another control group of pigeons was allowed free-flight. When the pigeons from the three groups were released from two distant release sites at about six months of age post-fledging, the two control groups were found to be equally good at orientating and returning home, while the experimental pigeons held in the shielded aviary for the first three months post-fledging were unable to orientate homeward and they were generally unsuccessful in returning home. This result supports the hypothesis that environmental experience during the first three months post-fledging is critical for some aspect of navigational map learning and that navigational map learning displays sensitive period-like properties.


Challenges of implementing the integrated disease surveillance and response strategy in Zambia: a health worker perspective.
Despite advances in medical technology and public health practice at the global level over the past millennia, infectious diseases are still the leading causes of death in most resource limited countries. Stronger infectious disease surveillance and response systems in developed countries facilitated the near elimination of infectious disease related deaths in those countries. Today, low-income countries are following this path by strengthening disease surveillance and response strategies that would help reverse the trend in infectious disease associated morbidity and mortality cases. In 2000, Zambia adopted the World Health Organisation Regional Office for Africa's (WHO-AFRO) Integrated Disease Surveillance and Response Strategy (IDSR) to monitor, prevent and control priority notifiable infectious diseases in the country. Through this strategy, activities pertaining to disease surveillance are coordinated and streamlined to take advantage of similar surveillance functions, skills, resources and targeted populations. The purpose of the study was to investigate and report on the existing challenges in the implementation of the IDSR strategy in a resource limited country from a health worker perspective. A qualitative study approach was used to achieve the study aim. Data was collected through key informant interviews with selected persons at the Lusaka Province Health Office (LPHO); Lusaka and Chongwe District Health Management Team Offices; and four selected health facilities in the two districts (two from each). Thematic analysis approach was used to analyse the qualitative data. The major successes included operationalised response and epidemic preparedness at all levels (National to district); full-time staff and budget dedicated to disease surveillance at all levels and adoption of the 2010 World Health Organisations' Integrated Disease Surveillance and Response Strategy technical guidelines to the Zambian context. Several challenges hampered effective implementation. These include inadequate trained human resources, poor infrastructure and coordination challenges. The implementation of IDSR strategy in Zambia has recorded some successes. However, several gaps hinder effective implementation. It is imperative that these gaps are addressed for Zambia to have a robust surveillance system that could inform policy in a comprehensive and timely manner.


Correlation between clinical nodal status and sentinel lymph node biopsy false negative rate after neoadjuvant chemotherapy.
Neoadjuvant chemotherapy (NAC) is the standard treatment for locally advanced breast cancer. It is now being used to treat operable breast cancer to facilitate breast-conserving surgery, but the accuracy of sentinel lymph node biopsy (SLNB) in breast cancer patients receiving NAC remains open to considerable debate. We enrolled 96 patients with stage II-III breast cancer who received NAC from January 2001 to July 2010. All patients underwent breast surgery and SLNB, followed immediately by complete axillary lymph node dissection (ALND). Sentinel lymph nodes were detected with blue dye and radiocolloid injected intradermally just above the tumor and then evaluated with hematoxylin and eosin and immunohistochemical staining. The overall identification rate for SLNB was 87.5% (84/96); the false negative rate (FNR) was 24.5% (12/49); and the accuracy rate was 85.7% (72/84). The FNR was significantly lower in clinically node-negative patients than in node-positive patients before NAC (5.5% vs. 35.5%; p=0.001). Accuracy was also significantly higher in clinically node-negative patients than in node-positive patients before NAC (97.2% vs. 77.1%; p=0.009). The FNR was 27.3% among 46 clinically node-positive patients before NAC who were clinically node-negative after NAC. Among 12 patients with a complete tumor response (CR), the FNR was 0%, compared with 26.1% for 83 patients with a partial response and stable disease (p=0.404). Although associated with a high FNR after NAC, SLNB would have successfully replaced ALND in clinically node-negative patients before NAC and in patients with a CR after NAC.


Vapor Pressure of Ice Containing D2O.
The vapor pressures of solid D(2)O and ice containing D2O in various amounts were measured with an oil manometer. The result gives the lowest vapor pressure for D(2)O ice ever obtained. The observed value for ice with different proportions of D2O and H(2)O agreed well with the values calculated on the assumption that the vapor pressure of HDO is the geometrical mean of those of H(2)O and D(2)O and that the equilibrium constant of isotopic exchange is 4. The fractionation factor of D between ice and vapor phases increases from 1.128 at 0 degrees C to 1.210 at -38 degrees C.


A Drosophila model of FUS-related neurodegeneration reveals genetic interaction between FUS and TDP-43.
Amyotrophic lateral sclerosis (ALS) is a late-onset neurodegenerative disorder characterized by the loss of motor neurons. Fused in sarcoma/translated in liposarcoma (FUS/TLS) and TAR DNA-binding protein (TDP)-43 are DNA/RNA-binding proteins found to be mutated in sporadic and familial forms of ALS. Ectopic expression of human ALS-causing FUS/TLS mutations in Drosophila caused an accumulation of ubiquitinated proteins, neurodegeneration, larval-crawling defect and early lethality. Mutant FUS/TLS localized to both the cytoplasm and nucleus, whereas wild-type FUS/TLS localized only to the nucleus, suggesting that the cytoplasmic localization of FUS/TLS is required for toxicity. Furthermore, we found that deletion of the nuclear export signal strongly suppressed toxicity, suggesting that cytoplasmic localization is necessary for neurodegeneration. Interestingly, we observed that FUS/TLS genetically interacts with TDP-43 in a mutation-dependent fashion to cause neurodegeneration in vivo. In summary, we demonstrate that ALS-associated mutations in FUS/TLS cause adult-onset neurodegeneration via a gain-of-toxicity mechanism that involves redistribution of the protein from the nucleus to the cytoplasm and is likely to involve an interaction with TDP-43.


First on-sky results of the CO-SLIDAR C(2)(n) profiler.
COupled SLope and scIntillation Detection And Ranging (CO-SLIDAR) is a recent profiling method of the vertical distribution of atmospheric turbulence strength (C(2)(n) profile). It takes advantage of correlations of slopes and of scintillation, both measured with a Shack-Hartmann wavefront sensor on a binary star. In this paper, we present the improved CO-SLIDAR reconstruction method of the C(2)(n) profile and the first on-sky results of the CO-SLIDAR profiler. We examine CO-SLIDAR latest performance in simulation, taking into account the detection noise bias and estimating error bars along with the turbulence profile. The estimated C(2)(n) profiles demonstrate the accuracy of the CO-SLIDAR method, showing sensitivity to both low and high altitude turbulent layers. CO-SLIDAR is tested on-sky for the first time, on the 1.5 m MeO (Métrologie Optique) telescope at Observatoire de la Côte d'Azur (France). The reconstructed profiles are compared to turbulence profiles estimated from meteorological data and a good agreement is found. We discuss CO-SLIDAR's contribution in the C(2)(n) profilers' landscape and we propose some improvements of the instrument.


In Tanzania, the many costs of pay-for-performance leave open to debate whether the strategy is cost-effective.
Pay-for-performance programs in health care are widespread in low- and middle-income countries. However, there are no studies of these programs' costs or cost-effectiveness. We conducted a cost-effectiveness analysis of a pay-for-performance pilot program in Tanzania and modeled costs of its national expansion. We reviewed project accounts and reports, interviewed key stakeholders, and derived outcomes from a controlled before-and-after study. In 2012 US dollars, the financial cost of the pay-for-performance pilot was $1.2 million, and the economic cost was $2.3 million. The incremental cost per additional facility-based birth ranged from $540 to $907 in the pilot and from $94 to $261 for a national program. In a low-income setting, the costs of managing the program and generating and verifying performance data were substantial. Pay-for-performance programs can stimulate the generation and use of health information by health workers and managers for strategic planning purposes, but the time involved could divert attention from service delivery. Pay-for-performance programs may become more cost-effective when integrated into routine systems over time.


Characteristics, comorbidities, and correlates of atypical depression: evidence from the UK Biobank Mental Health Survey.
Depression is a heterogeneous disorder with multiple aetiological pathways and multiple therapeutic targets. This study aims to determine whether atypical depression (AD) characterized by reversed neurovegetative symptoms is associated with a more pernicious course and a different sociodemographic, lifestyle, and comorbidity profile than nonatypical depression (nonAD). Among 157 366 adults who completed the UK Biobank Mental Health Questionnaire (MHQ), N = 37 434 (24%) met the DSM-5 criteria for probable lifetime major depressive disorder (MDD) based on the Composite International Diagnostic Interview Short Form. Participants reporting both hypersomnia and weight gain were classified as AD cases (N = 2305), and the others as nonAD cases (N = 35 129). Logistic regression analyses were conducted to examine differences between AD and nonAD in depression features, sociodemographic and lifestyle factors, lifetime adversities, psychiatric and physical comorbidities. Persons with AD experienced an earlier age of depression onset, longer, more severe and recurrent episodes, and higher help-seeking rates than nonAD persons. AD was associated with female gender, unhealthy behaviours (smoking, social isolation, low physical activity), more lifetime deprivation and adversity, higher rates of comorbid psychiatric disorders, obesity, cardiovascular disease (CVD), and metabolic syndrome. Sensitivity analyses comparing AD persons with those having typical neurovegetative symptoms (hyposomnia and weight loss) revealed similar results. These findings highlight the clinical and public health significance of AD as a chronic form of depression, associated with high comorbidity and lifetime adversity. Our findings have implications for predicting depression course and comorbidities, guiding research on aetiological mechanisms, planning service use and informing therapeutic approaches.