Regulating the suppressors: apoptosis and inflammation govern the survival of tumor-induced myeloid-derived suppressor cells (MDSC).
Immune suppressive myeloid-derived suppressor cells (MDSC) are present in most cancer patients where they inhibit innate anti-tumor immunity and are a significant obstacle to cancer immunotherapy. Inflammation is a known inducer of Gr1(+)CD11b(+) MDSC; however, the factors/conditions that regulate MDSC survival and half-life have not been identified. We have used mass spectrometry (MS) and proteomic analysis to identify proteins and pathways that regulate MDSC survival. This analysis revealed high expression of caspase family proteins and the Fas-FasL, p38 MAPK, and TGFβ pathways, suggesting that Fas-FasL apoptosis regulates MDSC survival. Flow cytometry, confocal microscopy, and western blot analyses confirmed the MS findings and demonstrated that Fas(+) MDSC are susceptible to Fas-mediated killing in vitro. In vivo studies with FasL-deficient and Fas-deficient mice demonstrated that Fas-FasL interactions are essential for MDSC apoptosis and for rejection of established metastatic disease and survival and that FasL(+) T cells are the effector population mediating MDSC apoptosis. MS findings validated by biological experiments demonstrated that inflammation increases MDSC levels by protecting MDSC from Fas-mediated apoptosis, possibly by activating p38 MAPK. These results demonstrate that MDSC half-life in vivo is regulated by FasL(+) T cells and that inflammation increases MDSC levels by conferring resistance to Fas-mediated apoptosis and identifies T cells as the relevant effector cells causing MDSC apoptosis in vivo. This newly recognized mechanism for regulating MDSC levels identifies potential new targets for decreasing MDSC in cancer patients.


Effects of interleukin 4 on CD25+CD4+ regulatory T cell function.
CD25+CD4+ regulatory T cells (Tregs) contribute to the maintenance of peripheral tolerance against self and non-self. The modulatory effects of cytokines, such as interleukin 4 (IL-4) on the function of Tregs have not been explored in detail. We here report that IL-4 prevents spontaneous apoptosis and the decline of foxp3 mRNA which were found to occur during culture of isolated Tregs. Tregs exposed to IL-4 were more potent in suppressing the proliferation of naïve CD4+ T cells and they better inhibited IFN-gamma production by CD4+ T cells as compared to Tregs cultured in medium. IL-4 also enhanced membrane IL-2Ralpha (CD25) expression on Tregs above the levels observed on freshly isolated cells. IL-4-mediated effects on Treg function persisted in Tregs from Stat6-/- mice, pointing to a Stat6-independent intracellular transduction pathway. In conclusion, our data suggest that the anti-inflammatory function of IL-4 could partly be mediated by effects on Tregs function.


Parenting and restrictions in childhood epilepsy.
From the overprotection literature, the predictive and interactional (moderation) effects of controlling and indulgent parenting on restrictions in children with epilepsy were examined. Parents of 73 children with epilepsy completed questionnaires on parenting, restrictions, and functional status. Predictive and moderation effects were tested using multiple regression analysis. Moderation was tested with interactive computational methods. Restrictions were significantly (R(2)=.38, FΔ=6.59***, p<.001) predicted from seizure frequency (β=.24*, p<.05), functional status (β=-.42***, p<.001), and interaction between controlling and indulgent parenting (β=.28**, p<.01). Moderation occurred predominantly for high values of control: controlling parents who were not indulgent imposed fewer restrictions. In contrast, controlling parents who were indulgent imposed more restrictions. Parents who were controlling and more indulgent imposed more restrictions. Clinicians should ask parents about parenting and restrictions. Future research should examine whether the current study's findings can be replicated.


Is erectile dysfunction related to self-esteem and depression? A prospective case-control study.
Sexual activity is important role in life of men. Decreased sexual function has a negative impact on the quality of life of the patients and their partners. In this study, we aimed to evaluate the relationship of erectile dysfunction (ED) with self-esteem, symptom severity and depression. The 80 patients were evaluated prospectively. Group 1 (cases group) included 40 patients who with ED an age range of up to 18-70 years. Group 2 (healthy control group) included 40 patients who apply to the urology clinic for various reasons without ED. All patients were filled international index of erectile function-5 (IIEF-5), Beck Depression questionnaire (BDI-2) and Rosenberg self-esteem questionnaires (RESQ). Demographic characteristics and inventory results of the patients were compared between the two groups. Significant differences were found between the two groups in IIEF-5, RESQ and BDI-2 scores. In Group 1, IIEF-5 score was low, RESQ, BDI-2 scores were significantly higher (All scores p < 0.001). As a result people significantly reflect their sexual satisfaction in their social life. The patients with mild ED are not entirely satisfied in spite of normal sexual frequency. This situation causes significantly low the self-esteem of men. Prevention of ED will contribute to increased self-esteem and happy lives.


Clinical efficacy of amino bisphosphonate on periodontal disease status in postmenopausal women: Randomized double-blind placebo-controlled trial.
Osteoporosis is a common skeletal disorder affecting postmenopausal women. Data suggest that postmenopausal women are at increased risk of periodontal diseases. Amino bisphosphonates are potent inhibitors of bone resorption and effectively used in the treatment of osteoporosis. Preliminary data indicate that there is a potential role for bisphosphonates in the management of periodontitis. Hence, this randomized placebo-controlled trial was designed to investigate the clinical efficacy of amino bisphosphonate on periodontal disease status among postmenopausal women. Thirty patients were randomly allocated to two treatment groups: Group A, which received scaling and root debridement and 70 mg weekly single oral dose of alendronate drug, and Group B, which received scaling and root debridement and placebo drug for 6 months. Clinical periodontal measurements were carried out for all patients at the baseline and 6 months later. Mandibular bone mineral density (BMD) was measured using a dual energy X-ray absorptiometer at the beginning of the study and the end of 6 months. A weekly single oral dose of 70 mg alendronate was well-tolerated. The intragroup comparison showed significant improvement in periodontal parameters in both groups. The intergroup comparison showed a significant increase in BMD after 6 months in Group A when compared with Group B (P = 0.0179). Single oral dose of 70 mg alendronate per week is well-tolerable, gastro-intestinally safe, and improves the clinical outcome of nonsurgical periodontal therapy.


Accuracy of intravitreal injection volume for aflibercept pre-filled syringe and BD Luer-Lok one-milliliter syringe.
To evaluate the accuracy of intravitreal injection volume of the pre-filled syringe (PFS) in which aflibercept is packaged compared to the BD Luer-Lok 1-mL syringe. Ophthalmologists injected their typical intravitreal volume for aflibercept using either the PFS or BD Luer-Lok 1-mL syringe for 5 times each. The injected fluid was weighed using a micro-scale and converted to volume. The volume of fluid injected was also evaluated when the 0.05 mL line on the PFS was lined up to the tip or base of the dome-shaped plunger. Injection volume was measured for 12 physicians. The average injected fluid volume was 74.22 ± 15.87 µL for PFS and 53.42 ± 4.61 µL for the BD Luer-Lok 1-mL syringe (p < 0.0001). The average deviation in volume injected for the PFS was higher compared to the BD Luer-Lok 1-mL syringe (11.36 µL vs. 3.35 µL, p < 0.0001). When the PFS was lined up with the tip of the dome-shaped plunger at the 0.05-mL line, the average injected volume was 71.03% higher. The intravitreal injection volume and variability using the new PFS were significantly higher than the volume injected using the BD Luer-Lok 1-mL syringe previously used, potentially leading to higher rates of visually significant elevation of intraocular pressures.


Novel proteolytic microvesicles released from human macrophages after exposure to tobacco smoke.
Cigarette smoking damages the extracellular matrix in a variety of locations, leading to atherosclerotic plaque instability and emphysematous lung destruction, but the underlying mechanisms remain poorly understood. Here, we sought to determine whether exposure of human macrophages, a key participant in extracellular matrix damage, to tobacco smoke extract (TSE) induces the release of microvesicles (MVs; or microparticles) with proteolytic activity; the major proteases involved; and the cellular mechanisms that might mediate their generation. We found that MVs released from TSE-exposed macrophages carry substantial gelatinolytic and collagenolytic activities that surprisingly can be predominantly attributed to a single transmembrane protease of the matrix metalloproteinase (MMP) superfamily (namely, MMP14). Flow cytometric counts revealed that exposure of human macrophages to TSE for 20 hours more than quadrupled their production of MMP14-positive MVs (control, 1112 ± 231; TSE-induced, 5823 ± 2192 MMP14-positive MVs/μL of conditioned medium; means ± SEM; n = 6; P < 0.01). Our results indicate that the production of these MVs by human macrophages relies on a series of regulated steps that include activation of two mitogen-activated protein kinases (MAPKs, i.e., the Jun N-terminal kinase and p38 MAPK), and then MAPK-dependent induction and maturation of cellular MMP14, a remarkable accumulation of MMP14 into nascent plasma membrane blebs, and finally caspase- and MAPK-dependent apoptosis and apoptotic microvesicle generation. Proteolytically active MVs induced by tobacco smoke may be novel mediators of clinical important matrix destruction in smokers.


Risk of herpes zoster according to past history in the general population: The Japanese Shozu herpes zoster study.
Immunity is known to persist after vaccination for varicella zoster virus, but the duration of immunity in patients who develop herpes zoster (HZ) remains unknown. To investigate the association between a past history of HZ and its occurrence in the general population. The Shozu HZ (SHEZ) cohort study included data for 12 299 individuals aged ≥50 years with information on their HZ history. Cross-sectional and 3-year follow-up studies were carried out to analyze the associations between a history of HZ (yes <10 years, yes ≥10 years, no) and the proportion of positive varicella zoster virus skin test results (erythema diameter ≥5 mm) and the risk of HZ after adjusting for potential confounding factors including age, sex, body mass index, smoking status, sleep duration, and mental stress. The incidences of positive skin test results were 87.7% (470/536) for individuals with a history of HZ <10 years ago, 82.2% (396/482) for those with a history of HZ ≥10 years, and 80.2% (3614/4509) for those with no history of HZ. The multivariable odds ratios (95% confidence intervals) of erythema diameter ≥5 mm were 2.07 (1.57-2.73) and 1. 39 (1.08-1.80) for individuals with a history <10 years and ≥10 years ago, respectively, compared with no history. The corresponding multivariable hazard ratios of HZ were 0.54 (0.34-0.85) and 1.16 (0.83-1.61), respectively. A past history of HZ <10 years ago may reduce the occurrence of HZ.


Increased expression of pro-inflammatory cytokines at the fetal-maternal interface in bovine pregnancies produced by cloning.
A significant rate of spontaneous abortion is observed in cattle pregnancies produced by somatic cell nuclear transfer (SCNT). Major histocompatibility complex class I (MHC-I) proteins are abnormally expressed on the surface of trophoblast cells from SCNT conceptuses. MHC-I homozygous compatible (n = 9), homozygous incompatible (n = 8), and heterozygous incompatible (n = 5) pregnancies were established by SCNT. Eight control pregnancies were established by artificial insemination. Uterine and trophoblast samples were collected on day 35 ±1 of pregnancy, the expression of immune-related genes was examined by qPCR, and the expression of trophoblast microRNAs was assessed by sequencing. Compared to the control group, trophoblast from MHC-I heterozygous incompatible pregnancies expressed increased levels of CD28, CTLA4, CXCL8, IFNG, IL1A, IL2, IL10, IL12B, TBX21, and TNF, while GNLY expression was downregulated. The MHC-I homozygous incompatible treatment group expressed increased levels of IFNG, IL1A, and IL2 while the MHC-I homozygous compatible group did not differentially express any genes compared to the control group. In the endometrium, relative to the control group, MHC-I heterozygous incompatible pregnancies expressed increased levels of CD28, CTLA4, CXCL8, IFNG, IL10, IL12B, and TNF, while GATA3 expression was downregulated. The MHC-I homozygous incompatible group expressed decreased amounts of CSF2 transcripts compared with the control group but did not have abnormal expression of any other immune-related genes. MHC-I incompatible pregnancies had 40 deregulated miRNAs compared to control pregnancies and 62 deregulated microRNAs compared to MHC-I compatible pregnancies. MHC-I compatibility between the dam and fetus prevented an exacerbated maternal immune response from being mounted against fetal antigens.


Lesions associated with the plexus venosus subcutaneus collaris of pigeons with chlorophacinone toxicosis.
We describe lesions in 29 adult domestic pigeons (Columba livia) poisoned with chlorophacinone, an indandione anticoagulant rodenticide. Birds were found dead in the field and in dovecotes after a wide-area treatment against common voles (Microtus arvalis) based on the dispersion in the open field of wheat grain treated with 0.005% chlorophacinone. At necropsy, most pigeons showed crops full of red-colored wheat grain, extensive subcutaneous hematoma in neck and breast zones, and hemorrhages in lungs and the coelomic cavity. Chlorophacinone was determined in liver samples by high-performance liquid chromatography with ultraviolet detection, and the mean (range) concentration was 11.2 (1.48-50.1) microg/g. Pigeons have a venous subcutaneous plexus in the neck zone (plexus venosus subcutaneus collaris), which has been described as an erectile tissue with a thermoregulation function. This case report describes a relationship between the subcutaneous hemorrhagic lesions and the plexus venosus collaris.


Defibrination of normal human blood in vitro: a method giving a high recovery of untraumatized cells.
A new automatic apparatus is described for the defibrination of blood in vitro at laboratory temperature which results in only small (less than 10%) losses of white cells. Freshly-drawn blood is mixed smoothly at high speed (greater than 1200 rpm) and fibrin is removed rapidly as formed. The apparatus is designed to produce minimum mechanical trauma to the blood. Subsequent in vitro incubation of red cells, granulocytes and lymphocytes demonostrated their viability to be comparable with cells obtained by other methods of defibrination.


Investigation of sensorimotor dysfunction in Parkinson disease by resting-state fMRI.
Functional MRI has played a fundamental role in Parkinson's disease(PD) study. In this paper, we performed an independent component analysis (ICA) based on functional networks to reveal the intricate variations on the morphology and functional properties of brain. Our analysis aims at discovering the differences between PD patients with sensorimotor function impairment and normal controls(NC), thus helping to understand the coordination neurological function degeneration in PD objectively. We investigated the blood oxygen level dependent(BOLD) functional MRI obtained at a 3.0 T MRI scanner. 30 PD patients and 28 NC subjects underwent the scan in resting state. The signals of sensory and motor coordinative control areas in the sensorimotor, insula and cerebellum networks acquired by ICA(Independent Component Analysis)were applied to analyze the functional alterations. Specifically, intra-network analysis was performed with signals in local networks, and inter-network analysis was conducted by functional network connectivity (FNC) with signals across different networks. Two sample T test was carried out to detect the significant (p < 0.05, FDR p < 0.05) functional abnormality in PD patients. We identified an obvious increase in bilateral posterior insula, but decrease in bilateral cerebellum hemisphere, supplementary motor area(SMA) and precentral gyrus paracentral lobule of left postcentral gyrus. Besides, we found a significantly increased connection between independent component (IC) 13 which was located in right postcentral gyrus and cerebellum. Decreased connections were detected between sensory and motor cortex in sensorimotor network and between cerebellum and insula network by FNC analysis in PD patients as well. Parkinson's disease derives from the degeneration of the dopaminergic neurons in substantia nigra, and results in decreased secretion of inhibitory neurotransmitter. The significant differences between PD and NC groups in our research maybe explain the clinical manifestations of prominent bradykinesia and multiple extrapyramidal symptoms.


Temporomandibular joint reconstruction with a custom total temporomandibular joint prosthesis: use in the multiply operated patient.
The purpose of this article is to report the development and use of the custom Christensen prosthesis (Garrett modification) for total temporomandibular joint reconstruction, its indications, protocol, adjunctive treatment and evolution to present design.


Immunomodulation of intestinal macrophages by mercury involves oxidative damage and rise of pro-inflammatory cytokine release in the fresh water fish Channa punctatus Bloch.
Mercury and its compounds have been parts of widespread pollutants of the aquatic environment. The present study was designed to assess the effect of mercury on fish immune responses. Since the metal is absorbed by fish and passed up the food chain to other fish-eating species, it not only affects aquatic ecosystems but also humans through bioaccumulation. In the present study, it was found that innate immunity of the fresh water fish Channa punctatus Bloch. was significantly debilitated after a periods of exposure to a sub-lethal concentration of mercury (0.3 mg/L). After 7 days of exposure, phagocytosis, cell adhesion and intracellular killing activity were found to decrease significantly along with significant decreases in nitric oxide (NO) and myeloperoxidase (MPO) production from macrophages as compared to the control group indicating intracellular damages. Levels of pro-inflammatory cytokines like TNF-α and IL-6 were found to be significantly more in mercury treated groups than that of control group indicating inflammatory damage. This included significant ultrastructural changes like fragmented epithelium, lesions in mucosal foldings, degenerated mitochondria, reduction in the number of goblet cells and disoriented microvilli as evident from transmission electron micrographs.


Monoterpene pyridine alkaloids and phenolics from Scrophularia ningpoensis and their cardioprotective effect.
Scrophularianines A-C (1-3), three new unusual monoterpene pyridine alkaloids with cyclopenta [c] pyridine skeleton reported from the genus Scrophularia for the first time, together with 15 known compounds (4-18), were isolated from the extract of Scrophularia ningpoensis. Their structures were elucidated on the basis of extensive analyses of spectroscopic evidences. The biogenetic relationship between monoterpene pyridine alkaloids and iridoids was proposed preliminarily. The myocardial protective bioassay indicated that compounds 13 and 14 with a concentration of 10(-4)M exhibited significantly protective effect against H2O2-induced apoptosis in cardiomyocytes.


A population-based survey of headache and migraine in 22,809 adults.
A population-based epidemiological study of the prevalence of headache in a sample of 24,682 adults was carried out in Finland. Enquiries were made concerning the frequency of headache, migraine and characteristics of headache attacks in a questionnaire survey in 1981. Headache was classified as migraine, and headache without migrainous features. Possible somatic causes of headache were excluded by record-linkage with three nation-wide medical registries. The prevalence of both headache types depended on age and gender. Among women the overall prevalence of migraine was 10.1% with a maximum of 11.5% in the 40-49 age group. The prevalence rate among men was 2.5% and the female/male gender ratio was 4.0. Women with migraine had an average of 52.5 headache occasions during a year and men with migraine 61 headache occasions per year. At the individual level, headache without migrainous features was least invalidating. There were only 16.5 headache occasions a year among women and 11.5 among men. However, from a population perspective headache without migrainous features accounted for 46.5% of all headache occasions among women and for 60.8% among men. Thus, from the public health view-point, studies of "non-clinical" headache, e.g. headache without migrainous features should be emphasized.


[Acute cardiovascular reactions to cervical nerve root infiltration].
The frequency of cardiovascular adverse effects of cervical nerve root infiltration was investigated by means of a prospective study. The hemodynamic and respiratory parameters systolic and diastolic blood pressure, heart rate, respiratory rate and oxygen saturation were monitored continuously in 56 patients undergoing cervical spine nerve root infiltration (injection of 10 ml Lidocain 0.5 %, mean depth of injection 5 cm). The monitored time frame ranged from 5 minutes before to 10 minutes after its administration, an additional holter-monitoring began 1 day prior to the injection. Severe complications such as syncopes were not observed in any of the 56 patients, 4 patients developed presyncopes. None of the registered parameters showed a significant change. In a group of patients with known, pre-existing cardiovascular morbidity, no significant changes occurred either. No relevant cardiac arrhythmias were observed. The observed cardiovascular complications were not severe. Most probably, the observed reactions were vasavagal presyncopes. Supine positioning led to immediate recovery in all of these patients. A complete hemodynamic monitoring and the placement of an intravenous line do not seem to be absolute necessities in the routine cervical nerve root infiltration.


Accord insertion in the 5' flanking region of CYP6G1 confers nicotine resistance in Drosophila melanogaster.
What has driven the sweep of the Accord retrotransposon insertion allele of CYP6G1 in the natural populations of Drosophila melanogaster is unknown. Previous studies on the DDT selection hypothesis produced conflicting data. To reexamine the DDT selection hypothesis and search for alternative explanations, we conducted a series of correlation and genetic linkage experiments with eight D. melanogaster natural populations collected from California (CM1, CM2, CM3, and CM7) and Africa (AM2, AM3, AM4, AM7). Diagnostic PCR showed that CM1, CM2, CM7, and AM3 have the Accord insertion in the CYP6G1 locus, whereas the other four strains do not. RT-PCR analysis exhibits a 100% correlation between Accord insertion and CYP6G1 overexpression. However, among the four strains with Accord-mediated CYP6G1 overexpression only CM1 and CM7 are resistant to DDT, and the other two strains (CM2 and AM3), like the four Accord-free strains, are susceptible to DDT. By contrast, all the four strains with Accord-mediated CYP6G1 overexpression are resistant to nicotine, a plant allelochemical. Genetic crosses between DDT resistant and susceptible Accord-insertion strains, as well as crosses between Accord-insertion and Accord-free strains demonstrated that Accord insertion and CYP6G1 overexpression are genetically linked to nicotine resistance rather than DDT resistance. These results suggest that naturally-occurring allelochemicals such as nicotine are the initial driving force for the worldwide prevalence of the Accord insertion allele of CYP6G1 in D. melanogaster natural populations.


The influence of heterotopic ossification on functional status of hip joint following total hip arthroplasty.
The functional failure induced by heterotopic ossification (HO) following total hip arthroplasty (THA) was analyzed and correlated to the radiologic failure. From July 1997 to July 2001, 315 patients (345 hips) received THA indicated by a hypertrophic osteoarthritis of higher degree (Kellgren grade III, IV). All patients were irradiated prophylactically for prevention of HO on the evening before surgery with a 7-Gy single fraction. The patients' median age was 66.3 years. Radiologic failure was assessed by comparison of pre- and postoperative hip X-rays (immediately and 6 months after surgery). Analysis of radiographs was performed according to the Brooker Score. Clinical failure was appraised by measurement of passive range of motion (ROM) of the hip joint with a standard goniometer. The t-test was used for statistical analysis. 281 patients (81.5%) did not develop HO. HO of Brooker grade I or II was found in 58 patients (16.8%). Six patients (1.7%) developed HO Brooker grade III or IV. There was a significant negative correlation between the degree of radiologic and clinical failure. ROM differed significantly between patients with HO Brooker grade 0, I, II and patients with HO Brooker grade III, IV. Comparing the pre- and postoperative ROM, all patients with Brooker grade 0, I and II showed a significant improvement of flexion, internal and external rotation, abduction and adduction movement. Patients with HO Brooker grade III and IV showed no improvement of ROM in the postoperative follow-up. The development of HO following THA influences the physical function of the hip joint dependent on the degree of ossification. HO of lower degree (Brooker I, II) does not influence the clinical outcome, whereas HO of higher degree (Brooker III, IV) reduces the function of hip arthroplasty. Therefore, the purpose of a prophylactic therapy must be to reduce HO of higher degree.


Immunoaffinity in-tube solid phase microextraction coupled with liquid chromatography with fluorescence detection for determination of interferon α in plasma samples.
The present work describes a sensitive and specific automated immunoaffinity in-tube solid-phase microextraction coupled with liquid chromatography with fluorescence detection (LC-FD) method for the determination of interferon α2a in plasma samples for therapeutic drug monitoring. To this end, the intrinsic selectivity of the monoclonal antibodies has been combined with in-tube solid phase microextraction by immobilization of these antibodies into the fused silica capillary. The in-tube SPME variables, such as plasma sample volume, draw/eject volume, number of draw-eject cycles, as well as desorption procedure have been optimized, in order to improve the sensitivity of the proposed method. Analyses of interferon α2a in plasma samples were carried out within 12min (sample preparation and LC analyses). The response of the proposed method was linear over a dynamic range 0.006-3.0MIUmL(-1), with a correlation coefficient of 0.998. The inter-day precision of the method had a coefficient of variation lower than 6.2%. The proposed automated method has adequate analytical sensitivity and selectivity for the determination of interferon α2a in plasma samples for therapeutic drug monitoring. The proposed method was successfully applied to the plasmas samples analyses from patients in therapy with interferon α-2a, demonstrating a rare application of in-tube SPME for biopharmaceutical (protein) analyses.


Biofertilization containing Paenibacillus triticisoli BJ-18 alters the composition and interaction of the protistan community in the wheat rhizosphere under field conditions.
Most studies focus on the effects of biofertilizer on the bacterial and fungal communities, and we still lack an understanding of biofertilizer on the protistan community. Here, the effects of biofertilizer containing Paenibacillus triticisoli BJ-18 on composition and interaction of the protistan community in the wheat rhizosphere were investigated. Biofertilizer application altered soil physicochemical properties and the protistan community composition, and significantly induced an alpha diversity decline. Random forecast and redundancy analysis demonstrated that nitrogenase activity and available phosphorus were the main drivers. Trichomonas classified to the phylum Metamonada was enriched by biofertilizer, and was significantly positive connected with soil nitrogenase activity and some function genes involved in nitrogen-fixation and nitrogen-dissimilation. Biofertilization loosely connected biotic interactions, while it did not affect the stability of the protistan community. Besides, biofertilizer promoted the connections of protists with fungi, bacteria, and archaea. Combined with biotic networks (protists, fungi, bacteria, and archaea) and interactions between protists and soil physicochemical properties/function genes, protists may act as keystone taxa potentially driving soil microbiome composition and function. Overall, these results suggest that the biofertilizer is a driver of the soil protistan community, contributing to ecosystem functioning.


Profiling glycoprotein n-linked oligosaccharide by capillary electrophoresis.
A method for analysis of N-linked oligosaccharides derived from glycoproteins including sialic acid-containing species is presented. It is based on the combination of specific chemical and enzymatic conversions coupled with capillary electrophoretic (CE) separation and laser-induced fluorescence (LIF) detection. Glycoproteins were heat-denatured in the presence of a reducing agent and the N-linked oligosaccharides were released by peptide N-glycosidase (PNGase F; EC220.127.116.11)-catalyzed hydrolysis. The released N-linked oligosaccharides were derivatized with 8-aminopyrene-1,3,6-trisulfonate (APTS) under mild reductive amination conditions in which desialylation and loss of fucose residues are minimized. A model N-linked oligosaccharide, desialylated, galactosylated biantennary, core-substituted with fucose (A2F) was tested for APTS-based derivatization chemistry with excellent recovery of the adduct without losing fucose and neuraminic acid residues. The profiles of heavily sialylated N-linked oligosaccharides derived from fetuin, recombinant human erythropoietin and kallikrein are reported and the data show that the present method produces a high resolution of the N-linked oligosaccharide profile for fingerprinting glycans derived from glycoproteins.


Hormonal regulation of inflammatory cell cytokine transcript and bioactivity production in response to endotoxin.
Tumor necrosis factor (TNF), interleukin 1 (IL-1) and interleukin 6 (IL-6) are central mediators of the inflammatory response. We investigated the modulation of these cytokines by hormones in vitro. Murine adherent peritoneal exudate cells (PEC) were exposed to various concentrations of hormones followed by lipopolysaccharide (LPS, 10 micrograms/ml). TNF, IL-1 and IL-6 production were assessed by bioassays, enzyme-linked immunosorbent assays (ELISA) or Western blot, and specific RNA transcripts by Northern blot. Hydrocortisone in concentrations as low as 10 ng/ml had dramatic inhibitory effects on supernatant levels of TNF and IL-1 and on TNF, IL-1 and IL-6 transcript number. Supernatant levels of IL-6 were only slightly diminished by hydrocortisone. Adrenocorticotrophic hormone (ACTH) and insulin increased supernatant levels of TNF bioactivity in response to LPS, while each decreased available TNF-alpha gene transcripts. Thus TNF protein production was affected at a post-transcriptional level. ACTH and insulin increased supernatant levels of IL-6 produced in response to LPS without altering available transcripts. Corticotrophin-releasing factor (CRF), epinephrine and glucagon had no effect on supernatant levels of cytokine. Thus, physiological and pharmacological concentrations of hydrocortisone had dramatic inhibitory effects on the supernatant levels of TNF and IL-1, and on the number of available TNF, IL-1 and IL-6 transcripts in PEC exposed to LPS, but had minimal effects on supernatant levels of IL-6 bioactivity. This hydrocortisone action may be a specific negative feedback system for IL-1 and TNF, with relative sparing of IL-6.


Impact of male obesity on infertility: a critical review of the current literature.
To evaluate the current understanding of the effects and potential mechanisms of obesity on male fertility. Literature review of articles pertaining to obesity and male infertility. Recent population-based studies suggest an elevated risk for subfertility among couples in which the male partner is obese and an increased likelihood of abnormal semen parameters among heavier men. Male factor infertility is associated with a higher incidence of obesity in the male partner. Obese men exhibit reduced androgen and SHBG levels accompanied by elevated estrogen levels. Reduced inhibin B levels correlate with degree of obesity and are not accompanied by compensatory increases in FSH. This complexly altered reproductive hormonal profile suggests that endocrine dysregulation in obese men may explain the increased risk of altered semen parameters and infertility. Additional features of male obesity that may contribute to an increased risk for infertility are altered retention and metabolism of environmental toxins, altered lifestyle factors, and increased risks for sexual dysfunction. Neither reversibility of obesity-associated male infertility with weight loss nor effective therapeutic interventions have been studied yet. The increasing prevalence of obesity calls for greater clinician awareness of its effects on fertility, better understanding of underlying mechanisms, and eventually avenues for mitigation or treatment.


Masseter muscle contraction and cervical muscle sensitization by nerve growth factor cause mechanical hyperalgesia in masticatory muscle with activation of the trigemino-lateral parabrachial nucleus system in female rats.
To establish a new rat model of craniofacial myalgia, and to clarify which central nervous system pathways are activated in the model. Craniofacial myalgia, represented by myogenous temporomandibular disorder and tension-type headache with pericranial tenderness, is more common in female patients. The pain is thought to be a type of multifactorial disorder with several coexisting causes. To our knowledge, there are no models of craniofacial muscle hyperalgesia caused by multiple types of stimuli. We injected nerve growth factor into the trapezius muscle of female and male rats and repeatedly stimulated the masseter muscle (MM) electrically for 10 days. We determined the mechanical head-withdrawal threshold of MM and extent of phosphorylated extracellular signal-related kinase 1/2 (pERK) immunoreactivity in various regions of the lower brainstem. We conducted retrograde tract-tracing to determine the projection of mechanosensitive MM-innervating secondary neurons to the lateral parabrachial nucleus. Finally, we administered morphine in rats to determine whether increases of pERK immunoreactivity were dependent on noxious inputs. In female rats, but not male rats, the mechanical head-withdrawal threshold was decreased significantly from days 9 to 12. The number of pERK-immunoreactive neurons in the brainstem was increased significantly in female rats in the group with both stimuli compared to rats in other groups with a single stimulus. Mechanosensitive MM-innervating neurons in the brainstem projected to the parabrachial nucleus. Morphine administration blocked the increase in the number of pERK-immunoreactive neurons in both the brainstem and parabrachial nucleus. We established a model of craniofacial myalgia by combining trapezius and MM stimuli in female rats. We found mechanical hyperalgesia of the MM and activation of the pain pathway from the brainstem to parabrachial nucleus. The model reflects the characteristics of patients with craniofacial myalgia and might be helpful to clarify the pathogenic mechanisms underlying these disorders.


Purification and kinetic and structural properties of spinach leaf NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase.
NADP-dependent nonphosphorylating D-glyceraldehyde-3-phosphate dehydrogenase (EC 220.127.116.11) from spinach leaves has been purified to apparent electrophoretic homogeneity by ammonium sulfate fractionation, molecular sieving on Sephadex G-200, DEAE-cellulose, and 2',5'-ADP-Sepharose affinity chromatography. The purified enzyme exhibited a specific activity of 15 mumol (mg protein)-1 min-1 and was characterized as a homotetramer with a native molecular weight of 195,000. Preincubation of the purified enzyme with NADP+ resulted in an almost twofold increase in enzymatic activity. The rate of activation was slower than the rate of catalysis, indicating that the enzyme has hysteretic properties. This behavior results in a lag phase during activity measurement of the enzyme preincubated without NADP+. Substrate interaction and product inhibition studies suggest a rapid equilibrium random BiBi mechanism for the reaction. Thiol modifying reagents, iodoacetamide and diamide, completely inactivated the purified enzyme. Inactivation by iodoacetamide exhibited pseudo-first-order kinetics with a rate constant of 0.17 min-1. D-Glyceraldehyde 3-phosphate effectively protected the enzyme against inactivation by thiol reagents, suggesting that modification occurred at or near the substrate-binding site. Complete inactivation of the dehydrogenase was correlated with incorporation of 8 mol [1-14C]iodoacetamide/mol enzyme. Total protection afforded by D-glyceraldehyde 3-phosphate against enzyme inactivation by iodoacetamide was correlated with a protection of 4 mol reactive residues/mol enzyme. On the basis of these results it is suggested that one sulfhydryl group per enzyme subunit is essential for catalysis in spinach leaf nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase. A kinetic and molecular mechanism for the reaction is proposed.


H(+)-ATPase and transport of DOPAC, HVA, and 5-HIAA in monoamine neurons.
The effects of N-methylmaleimide (N-MtM), a vacuolar H(+)-ATPase inhibitor, were evaluated in the putamen of the cat to study the in vivo transport mechanisms of dopamine (DA), 5-hydroxytryptamine (5-HT), and their metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindolacetic acid (5-HIAA), using the brain focal microdialysis technique combined with HPLC. The addition of N-MtM to the perfusate altered invariably the flux of the DOPAC, HVA, and 5-HIAA in a similar pattern, resulting in a decrease of the extracellular levels of such metabolites, its extent being N-MtM concentration dependent, thus indicating that the mechanism(s) of such a decrease is (are) related most likely to decreased transport from the intracellular to the extracellular space as the consequence of the inhibition of the vacuolar H(+)-ATPase of DA and 5-HT neurons by the N-MtM. Furthermore, N-MtM masked the release of DA and 5-HT produced by KCl 120 mmol/l. Indeed, N-MtM increased the extracellular levels of such transmitters to values exceeding 4 to 6 times of those produced by KCl 120 mmol/l alone, which suggests that vacuolar H(+)-ATPase is probably involved also in the retention and/or reuptake process of DA and 5-HT.


Alteration in the brain content of substance P (1-7) during withdrawal in morphine-dependent rats.
Previous studies have shown that substance P (SP) may modulate the abstinence reaction to opioid withdrawal. Its N-terminal fragment SP1-7 may inhibit the intensity of the withdrawal reactions in morphine dependent mice. This study was designed to determine whether the endogenous concentrations of the SP1-7 fragment in the brain are affected during naloxone-precipitated withdrawal in the male rat. The amounts of the peptide was assessed by a specific radioimmunoassay in extracts of discrete brain regions (including the cerebral cortex, hippocampus, hypothalamus, nucleus accumbens, striatum, substantia nigra, ventral tegmental area and the spinal cord) during morphine tolerance and withdrawal. The results indicated that the concentrations of SP1-7 were significantly elevated in the ventral tegmental area both in morphine tolerant rats and during naloxone-precipitated withdrawal. During morphine withdrawal significant increases in the peptide concentration were also observed in the hypothalamus and the spinal cord. It was concluded that the enhanced content of SP1-7 may also indicate the involvement of the SP system during opioid withdrawal in the rat. The enhanced production of SP1-7 may reflect an increased release and/or metabolism of SP, which, in turn, counteracts the withdrawal.


PKD-dependent PARP12-catalyzed mono-ADP-ribosylation of Golgin-97 is required for E-cadherin transport from Golgi to plasma membrane.
Adenosine diphosphate (ADP)-ribosylation is a posttranslational modification involved in key regulatory events catalyzed by ADP-ribosyltransferases (ARTs). Substrate identification and localization of the mono-ADP-ribosyltransferase PARP12 at the trans-Golgi network (TGN) hinted at the involvement of ARTs in intracellular traffic. We find that Golgin-97, a TGN protein required for the formation and transport of a specific class of basolateral cargoes (e.g., E-cadherin and vesicular stomatitis virus G protein [VSVG]), is a PARP12 substrate. PARP12 targets an acidic cluster in the Golgin-97 coiled-coil domain essential for function. Its mutation or PARP12 depletion, delays E-cadherin and VSVG export and leads to a defect in carrier fission, hence in transport, with consequent accumulation of cargoes in a trans-Golgi/Rab11-positive intermediate compartment. In contrast, PARP12 does not control the Golgin-245-dependent traffic of cargoes such as tumor necrosis factor alpha (TNFα). Thus, the transport of different basolateral proteins to the plasma membrane is differentially regulated by Golgin-97 mono-ADP-ribosylation by PARP12. This identifies a selective regulatory mechanism acting on the transport of Golgin-97- vs. Golgin-245-dependent cargoes. Of note, PARP12 enzymatic activity, and consequently Golgin-97 mono-ADP-ribosylation, depends on the activation of protein kinase D (PKD) at the TGN during traffic. PARP12 is directly phosphorylated by PKD, and this is essential to stimulate PARP12 catalytic activity. PARP12 is therefore a component of the PKD-driven regulatory cascade that selectively controls a major branch of the basolateral transport pathway. We propose that through this mechanism, PARP12 contributes to the maintenance of E-cadherin-mediated cell polarity and cell-cell junctions.


Acrylamide stimulates glutamine uptake in Fischer 344 rat astrocytes by a mechanism involving upregulation of the amino acid transport system N.
High demand of neoplastic tissues for glutamine (Gln) is met by its active transport across cell membranes. Chronic treatment with acrylamide in rodents is associated with an increased incidence of neoplasms, including astrocytomas. In this study, 24-h acrylamide treatment significantly increased the initial rate of l-[G-3H]glutamine uptake in astrocyte cultures derived from the acrylamide-sensitive Fischer 344 rat, and this effect could be fully inhibited by histidine, a model substrate for the amino acid transport system N. RT-PCR analysis revealed that acrylamide treatment caused a significant increase in the astrocytic expression of the mRNA coding for the major system N protein, SNAT3, which is specifically overexpressed in malignant gliomas in situ. The acrylamide-induced upregulation of astrocytic Gln transport via system N is likely to affect Gln homeostasis in these cells and may be causally related to the increased astrocytoma incidence observed in Fischer 344 rats.


Standardization of 134Cs by three methods.
The nuclide 34Cs decays by beta-emission followed by gamma-deexcitation to 134Ba with a half-life T(1/2) = 2.065 a. It has been standardized by three methods: liquid scintillation counting (LSC), 4pi beta-gamma coincidence counting and 4pi gamma counting. In the LSC measurements, the CIEMAT/NIST method was used to calculate the efficiency. For the coincidence measurements, a conventional 4pi beta (proportional counter)-gamma(NaI) system was used. For the 4pi gamma standardization, a well-type Nal(Tl) detector was modeled with the Monte Carlo package PENELOPE, and the counting efficiency obtained by calculation. Results of the three methods agree within 0.65%.


Identification and Characterization of a Dominant Sulfolane-Degrading Rhodoferax sp. via Stable Isotope Probing Combined with Metagenomics.
Sulfolane is an industrial solvent and emerging organic contaminant affecting groundwater around the world, but little is known about microbes capable of biodegrading sulfolane or the pathways involved. We combined DNA-based stable isotope probing (SIP) with genome-resolved metagenomics to identify microorganisms associated with sulfolane biodegradation in a contaminated subarctic aquifer. In addition to 16S rRNA gene amplicon sequencing, we performed shotgun metagenomics on the 13C-labeled DNA to obtain functional and taxonomic information about the active sulfolane-degrading community. We identified the primary sulfolane degrader, comprising ~85% of the labeled community in the amplicon sequencing dataset, as closely related to Rhodoferax ferrireducens strain T118. We obtained a 99.8%-complete metagenome-assembled genome for this strain, allowing us to identify putative pathways of sulfolane biodegradation. Although the 4S dibenzothiophene desulfurization pathway has been proposed as an analog for sulfolane biodegradation, we found only a subset of the required genes, suggesting a novel pathway specific to sulfolane. DszA, the enzyme likely responsible for opening the sulfolane ring structure, was encoded on both the chromosome and a plasmid. This study demonstrates the power of integrating DNA-SIP with metagenomics to characterize emerging organic contaminant degraders without culture bias and expands the known taxonomic distribution of sulfolane biodegradation.


Terbium chelation, a specific fluorescent tagging of human transferrin. Optimization of conditions in view of its application to the HPLC analysis of carbohydrate-deficient transferrin (CDT).
Transferrin (Tf) is the major iron-transporting protein in the human body and, for this reason, has been extensively studied in biomedicine. This protein undergoes a complex glycosylation process leading to several glycoforms, some of which are important in the diagnosis of alcohol abuse and of congenital glycosylation defects under the collective name of carbohydrate-deficient transferrin (CDT). Exploiting the Tf ability to bind not only iron but also other ions, specific attention has been devoted to binding activity towards Tb3+, which was reported to greatly enhance its intrinsic fluorescence upon the interaction with Tf. However, the structural properties of the Tb3+-Tf complex have not been described so far. In the present work, the formation of the Tf-Tb3+ complex has been investigated by the employment of several biophysical techniques, such as fluorescence resonance energy transfer (FRET), "native" mass spectrometry (MS), and near-UV circular dichroism (CD). Each method allowed the detection of the Tf-Tb3+ complex, yielding a specific signature. The interaction of Tb3+ with Fe3+-free Tf (apoTf) has been described in terms of stoichiometry, affinity, and structural effects in comparison with Fe3+. These experiments led to the first direct detection of the Tf-Tb3+ complex by MS, indicating a 1:2 stoichiometry and allowing the investigation of structural effects of metal binding. Either Tb3+ or Fe3+ binding affected protein conformation, inducing structural compaction to a similar extent. Nevertheless, near-UV CD and pH-dependence profiles suggested subtle differences in the coordination of the two metals by Tf side chains. Experimental conditions that promote complex formation have been identified, highlighting the importance of alkaline pH and synergistic ions, such as carbonate. On the basis of these studies, sample pretreatment, separation, and detection conditions of a high-performance liquid chromatographic method for CDT analysis are optimized, achieving relevant increase (by a factor of ∼3) of analytical sensitivity. Graphical abstract Schematic representation of HPLC-separated transferrin glycoforms detected by fluorescence emission of the terbium ions bound to the protein.


Reirradiation for rectal cancer and surgical resection after ultra high doses.
Local recurrence of rectal cancer following high-dose pelvic radiation presents a difficult management challenge. Conventional wisdom suggests that reirradiation should be avoided and radical pelvic surgery is hazardous after ultra high-dose radiation. In a unique Phase I/II pilot study, 32 patients with recurrent rectal cancers following previous pelvic radiation underwent planned reirradiation to the pelvis. Initial radiation doses had ranged from 30-64.87 Gy (median dose 45 Gy). Seventeen patients underwent reirradiation followed by radical resection. Fifteen patients were reirradiated for palliative relief of symptoms. Treatment techniques consisted of two lateral fields (7 x 7 to 12 x 10 cm) encompassing the tumor with 2 cm margins. Reirradiation doses ranged from 19.80-47.66 Gy, (median 34.2 Gy). Patients also received concurrent low-dose continuous infusion chemotherapy, (5-FU 200-300 mg/day). Total cumulative radiation doses ranged from 70.6 to 111.6 Gy. Treatment was well tolerated. Four patients had radiation interrupted/discontinued for diarrhea or leukopenia. Follow-up ranges from 6 months to 36 months. No late sequelae of radiation have been observed to date. Seventeen patients underwent surgical exploration 6-8 weeks following reirradiation. Two patients had extensive disease and were not resected. Fifteen patients underwent radical resection of residual tumor (4 posterior exenterations, 6 APR, 3 transanal abdominal transanal proctocolectomy with coloanal anastomosis (TAATA), and 2 LAR). No patients died postoperatively. No excessive edema, hemorrhage, or adhesions were observed. Two patients developed pelvic abscess and one developed a coloanal stricture. Eleven of 15 resected patients are alive from 6 to 36 months with a 2-year survival of 66%. Of the patients treated palliatively, symptomatic relief was observed in 13/15 patients. No objective complete response was observed, but 6/15 patients had measurable partial response. Median survival in this group was 14 months. Based on this experience, we believe that in selected patients radical surgical resection after cumulative ultra high doses (70-90 Gy) of radiation can be performed safely. A viable anastomosis is also possible in spite of these high doses. Planned reirradiation for palliative relief of symptoms can be effective without unusual risks of complication. Long-term effects of such ultra high dose radiation and surgery continue to be monitored.


Induced pluripotent stem cells as a discovery tool for Alzheimer׳s disease.
The ability to accurately and systematically evaluate the cellular mechanisms underlying human neurodegenerative disorders such as Alzheimer׳s disease (AD) should lead to advancements in therapeutics. Recent developments in human induced pluripotent stem cells (iPSCs) have afforded the opportunity to use human neurons and glia to study cellular changes involved in neurological diseases. iPSCs have the potential to be differentiated into AD-relevant cell types, including forebrain neurons, astrocytes, and microglia. This permits the evaluation of individual cell types in isolation or in concert, thus modeling the interdependence of cell types within the brain. When discussing the potential of modeling AD with iPSCs, it is important to remember that the umbrella diagnosis of "Alzheimer׳s disease" represents a disease that is heterogeneous in terms of age of onset, underlying causes, and at times precise pathology. The ability of iPSCs to be derived from an array of AD patients allows for a closer examination of the mechanism of disease progression in particular subsets of subjects, who may have different mutations and allelic variants affecting their risk for disease. Disease mechanisms can be probed both by the genetic manipulation of iPSCs and by modifications to the cellular environment by chemical treatment. These studies may lead not only to the refinement of known pathways implicated in AD, but also to the identification of novel pathways heretofore unaffiliated with disease pathology. In this review, we describe the potential of iPSC models to transform our understanding of AD and to lead to valuable advancements in therapeutics. This article is part of a Special Issue entitled SI: Exploiting human neurons.


Intracranial Nonskull-Based Chondrosarcoma Arising from the Sagittal Sinus: A Case Report and Review of the Literature.
Intracranial nonskull-based chondrosarcoma is a very rare malignant tumor. In fact, it is difficult to diagnose because of its atypical radiologic and morphologic features. We report a case of an intracranial parasagittal chondrosarcoma in the left parietal lobe accompanied with significant peritumoral edema (an extremely rare phenomenon for this tumor), with a review of the literature, to clarify aspects in the diagnosis of this rare tumor. A literature search was using PubMed with specific key terms, inclusion criteria, and exclusion criteria. Selected case studies and case series were then compared and summarized them in Table 1. A 59-year-old male patient presented with a history of progressive right hemiparesis and loss of sensation for 45 days. Magnetic resonance imaging revealed a 2.6 × 2.4 × 2.8-cm mass arising from the superior sagittal sinus in his left parietal lobe. A left parietal parasagittal craniotomy was performed, and a macroscopically complete excision of the tumor was achieved. The patient recovered well postoperatively. Regular follow-up after 6 months revealed that he had almost recovered full strength to his right limbs, and there was no evidence of recurrence. Intracranial parasagittal chondrosarcoma in the parietal lobe is extremely rare. Although magnetic resonance imaging was performed, it is sometimes difficult to distinguish intracranial chondrosarcoma from meningioma and glioma, especially when the tumor arises from a nonskull base such as the meninges. Some cases could have significant peritumoral edema, although it is extremely rare. Histologic examination may aid in the diagnosis of this tumor. Neurosurgery is the most effective therapy for these tumors. Postoperative radiotherapy needs to be considered when the tumor is incompletely resected, has atypical histology, or is associated with significant peritumoral edema.


Cancer prevention and detection. An overview of variables influencing adoption and practice.
This paper presents an overview of factors, attitudes, and beliefs that may influence the adoption of cancer prevention and detection strategies by health-care providers and the public. Health-care practitioners require education about prevention and detection approaches to health care and the social variations in the population that may influence acceptance and adoption of these measures. The need for consistent guidelines among health professional organizations is also stressed.


Insulin receptor signaling through non-tyrosine kinase pathways: evidence from anti-receptor antibodies and insulin receptor mutants.
Although there is general agreement that insulin receptor tyrosine kinase activity mediates many of the actions of insulin, two types of studies suggest that non-tyrosine kinase dependent pathways may also exist. First, both monoclonal and polyclonal antibodies to the receptor have been shown to mediate many of insulin's actions with little or no stimulation of receptor kinase. Second, insulin receptor mutants, with reduced or no tyrosine kinase activity, have been shown to mediate several actions of insulin. Non-tyrosine kinase pathways that could signal insulin effects through the insulin receptor include non-covalent activation of G proteins, phospholipase Cs, or docking proteins such as IRS-1. Further studies on the chemical structures of phospholipids and their hydrolysis products involved in insulin action will be required to sort out the underlying mechanisms of insulin action via non-tyrosine kinase dependent pathways.


Stability after mandibular setback: mandible-only versus 2-jaw surgery.
The objective of this study was to evaluate whether changes in the technique for mandibular setback surgery since the introduction of rigid internal fixation have improved postoperative stability in Class III correction with setback alone and 2-jaw surgery. Cephalometric (skeletal and dental) outcomes for 17 patients with mandibular setback alone were compared with outcomes in 83 patients with 2-jaw surgery for Class III correction. Demographic characteristics in the 2 groups were similar, and the mean amount of setback (-4.7 mm) was the same; however, given a mean maxillary advancement of 4.9 mm, the 2-jaw patients had a greater total Class III correction. Greater than 4 mm of posterior movement of the gonion at surgery and a resulting significant change in ramus inclination were found in 8 of the mandible-only patients (47%) but only 1 of the 2-jaw patients (1%). Postoperatively, the mean changes for the 2 groups were similar, with mean forward movement of the chin (pogonion) of 2.8 mm in both groups, but the mechanism was different. In the mandible-only patients, the major reason for forward movement of the chin was recovery of ramus inclination. In the 2-jaw group, about half the change in chin position was because of forward movement of the gonion; the other half was because of small upward movement of the maxilla that allowed upward-forward rotation of the mandible. In both groups there was a significant correlation (r = 0.42, P < .0001) between postoperative change in the position of the chin and gonion. Despite improvements in surgical techniques for mandibular setback since 1995, postoperative stability still leaves something to be desired, but there is better control of the ramus position when 2-jaw surgery is performed.


ACPA and bone loss in rheumatoid arthritis.
Rheumatoid arthritis (RA) is an autoimmune inflammatory disease characterized by bone loss. Degree of inflammation and autoantibody positivity have both been identified as important initiators of skeletal damage in RA. Whereas it is well appreciated that inflammation initiates bone loss via the action of cytokines, the effect of autoantibodies in initiating bone destruction has long been underestimated. It has, nonetheless, been known for years that antibodies against citrullinated proteins (ACPA) and rheumatoid factor are associated with a more destructive disease course. It was recently shown that ACPA bind osteoclast precursor cells and directly promote their differentiation into bone-resorbing osteoclasts. Other results have shown that in ACPA-positive individuals bone loss starts even before the onset of clinical disease; this is indicative of the independent effect of these antibodies in initiating skeletal damage. The observation that the mere presence of ACPA is associated with pathological changes suggests that these antibodies have functional properties and initiate the onset of disease long before patients consult the rheumatologist because of symptomatic joint disease. These findings also indicate that "RA" is a syndrome rather than a single disease, suggesting that autoantibody-positive patients are both genetically and clinically distinct phenotypes.


Co-existence of abdominal aortic aneurysms and intracranial aneurysms.
The occurrence of abdominal aortic aneurysms (AAAs) and intracranial aneurysms (IAs) in the same patient and in the same family was studied among 89 patients with AAAs and 485 patients with IAs. Among the AAA-patients two had IAs themselves and five had IAs in the family, whereas three IA-patients had AAAs themselves and eight had AAAs in the family. Moreover, one of the patients with both AAA and IA had a blood relative with AAA, and in six of the families with both types of aneurysms there were more than two subjects with aneurysms. The results indicate, that AAAs and IAs may have a common aetiologic factor.


Bio-catalysis of mango industrial waste by newly isolated Fusarium sp. (PSTF1) for pectinase production.
The dried mango fruit processing industrial waste (MIW) used as carbon source for the production of pectinase from fungal strains. Eight fungal strains were isolated from MIW and screened for their ability to produce pectinase by pectin clear zone (PCZ) technique. Fusarium sp. (PSTF1) showed highest PCZ value of 52 mm. The physico-chemical characteristics of the medium were standardized for high production of pectinase. The highest production of pectinase in submerged fermentation observed at temperature-28 °C, pH-6.0, inoculum-size 0.6/25 ml, incubation-72 h, substrate concentration-0.6 g/100 ml, carbon source-fructose (1 %). The effect of different amino acids, vitamins also observed. Under these optimal conditions the highest activity 81.9718 µmol/ml of pectinase was observed. The Fusarium sp. (PSTF1) has been considered as the best pectinase producer in submerged fermentation of MIW. The cheap waste raw material used as best carbon source for high production of high value pectinase.


Changes in adiponectin:leptin ratio among older adults with obesity following a 12-month exercise and diet intervention.
Excess adiposity is characterized by alterations in adipokine secretion such that circulating leptin concentrations are increased with reductions in adiponectin. An emerging biomarker for the assessment of this adipose tissue (AT) dysfunction is the adiponectin:leptin (AL) ratio. A low AL ratio may be suggestive of dysfunctional AT and, consequently, a heightened cardiometabolic disease risk. This ancillary study investigated the relationship between the AL ratio and cardiometabolic health among community-dwelling older adults with obesity, as well as the effects of a 12-month exercise and diet intervention on changes in the AL ratio. Participants (n = 163, 70.2 ± 4.7 years, 38.0% male) were randomized to the exercise only group, exercise + nutrient-dense weight maintenance group (exercise + weight maintenance), or exercise + nutrient-dense caloric restriction of 500 kcal/d group (exercise + intentional weight loss) (clinicaltrials.gov #NCT00955903). Total and regional adiposity as determined by magnetic resonance imaging (MRI) and dual-energy X-ray absorptiometry (DXA), anthropometrics, and cardiometabolic biomarkers were assessed at baseline and 12 months. The AL ratio was significantly (p < 0.05) inversely correlated with body mass index, waist circumference, measures of adiposity, and insulin among all participants at baseline. Among females only, significant positive and inverse correlations were also observed between this ratio and high-density lipoprotein cholesterol and the inflammatory biomarkers high sensitivity C-reactive protein and interleukin-6, respectively. While controlling for biological sex, a significant time by intervention group interaction effect (p < 0.05) was observed such that the AL ratio significantly increased from baseline to study completion among participants in the exercise + weight maintenance group and exercise + intentional weight loss group. Post hoc analysis revealed that the exercise + intentional weight loss group exhibited a significantly greater AL ratio at study completion compared to other groups (p < 0.05 all). Results are in support of the AL ratio as a measure of AT dysfunction among older adults. Furthermore, results suggest that a 12-month exercise and diet intervention with intentional weight loss assists in improving the AL ratio in this population.


[Oncogenes, anti-oncogenes and their alterations in human tumors].
This review summarizes the discovery of oncogenes which are the activated versions of normal cellular protooncogenes which serve essential functions in the control of cell proliferation and differentiation. Their activation disrupts the normal control mechanism which rests on the subtle balance between their effects and those of anti-oncogenes or tumor suppressor genes. One may reasonably hope that defining a complete profile of alterations of both these types of essential genes in the tumor or in the clinically healthy individual will be invaluable to assess prognosis or genetic predisposition, respectively.


The brain in vivo expresses the 2',3'-cAMP-adenosine pathway.
Although multiple biochemical pathways produce adenosine, studies suggest that the 2',3'-cAMP-adenosine pathway (2',3'-cAMP→2'-AMP/3'-AMP→adenosine) contributes to adenosine production in some cells/tissues/organs. To determine whether the 2',3'-cAMP-adenosine pathway exists in vivo in the brain, we delivered to the brain (gray matter and white matter separately) via the inflow perfusate of a microdialysis probe either 2',3'-cAMP, 3',5'-cAMP, 2'-AMP, 3'-AMP, or 5'-AMP and measured the recovered metabolites in the microdialysis outflow perfusate with mass spectrometry. In both gray and white matter, 2',3'-cAMP increased 2'-AMP, 3'-AMP and adenosine, and 3',5'-cAMP increased 5'-AMP and adenosine. In both brain regions, 2'-AMP, 3-AMP and 5'-AMP were converted to adenosine. Microdialysis experiments in 2',3'-cyclic nucleotide-3'-phosphodiesterase (CNPase) wild-type mice demonstrated that traumatic brain injury (controlled cortical impact model) activated the brain 2',3'-cAMP-adenosine pathway; similar experiments in CNPase knockout mice indicated that CNPase was involved in the metabolism of endogenous 2',3'-cAMP to 2'-AMP and to adenosine. In CSF from traumatic brain injury patients, 2',3'-cAMP was significantly increased in the initial 12 h after injury and strongly correlated with CSF levels of 2'-AMP, 3'-AMP, adenosine and inosine. We conclude that in vivo, 2',3'-cAMP is converted to 2'-AMP/3'-AMP, and these AMPs are metabolized to adenosine. This pathway exists endogenously in both mice and humans.


Acidolysis as a biorefinery approach to producing advanced bioenergy from macroalgal biomass: A state-of-the-art review.
Facing fossil fuels consumption and its accompanying environmental pollution, macroalgae, as a major part of the third-generation (3G) biomass, has great potential for bioenergy development due to its species-abundant, renewable and carbohydrate-rich properties. Diluted acid treatment is one of the most effective approaches to releasing fermentable sugars from macroalgal biomass in a short period, but the optimal conditions need to be explored to maximize the hydrolytic yield for the subsequent microbial conversion. Therefore, this review aims to summarize the latest advances in various acids and other auxiliary methods adopted to increase the hydrolytic efficiency of macroalgae. Following an overview of the strategies of different algal types, methods involved in the bioconversion of biofuels and microbial fuel cells (MFC) from algal hydrolysates are also described. For the 3G biorefinery development, the review further discusses key challenges and trends for future utilizing marine biomass to achieve the large-scale industrial production.


Research report: susto and pesticide poisoning among Florida farmworkers.
This paper discusses research designed to investigate farmworker interpretation of a serious incident of pesticide poisoning, specifically, whether this incident was considered to precipitate the Mexican folk illness "susto." The study revealed that a total of 23% of the population interviewed felt that they definitely or possibly had developed susto as a result of the pesticide exposure. These patients complained of more residual symptoms from the pesticide exposure than did those who had been affected by the poisoning but who did not feel they suffered from susto. The findings have implications for medical providers working with these populations. A self diagnosis of "susto" may indicate that a patient is indeed sicker than other indicators suggest, and point to a need for increased follow-up.


Comparative study with in situ hybridization and immunocytochemistry in detection of HIV-1 in formalin-fixed paraffin-embedded cell cultures.
The sensitivities of three immunohistological techniques were compared in this study for detecting human immunodeficiency virus (HIV-1) in infected cultured human lymphocytes that had been formalin-fixed and paraffin-embedded. The techniques included in situ hybridization (ISH) with HIV-1 cDNA; immunocytochemistry with HIV-1 p24 monoclonal antibody (ICC-m); and immunocytochemistry with HIV-1 polyclonal antibody from a patient with acquired immunodeficiency syndrome (AIDS) (ICC-p). Procedures were optimized for enzyme digestion and for antibody reaction conditions. HIV-1--infected cells and noninfected control cells were tested. Noninfected controls were uniformally negative by all three methods. Infected cells had the highest positivity rate by the ISH method (p less than or equal to 0.0001), and the ICC-p method was more positive than the ICC-m (p less than or equal to 0.0001). Both the ICC-p and the ICC-m techniques were more positive with the cocultivated cell cultures than the ISH, which was more sensitive with the infected continuous cell line (P less than or equal to 0.0001). The ICC-p method had a lower standard deviation on positive results than either the ICC-m or ISH method. The variability observed with these test procedures, reagents, and specimens suggests that these are important technological parameters in detecting p24, with implications for detecting other HIV-1 markers in infected tissues.


Gastric-tube esophagoplasty in children.
From 1979 to 1996, 19 patients underwent gastric-tube esophagoplasty. There were 10 boys and 9 girls, aged between 1 year 4 months and 4 years 11 months at the time of surgery. In 1 patient the esophagoplasty was performed due to a long stenosis secondary to reflux; 3 others had caustic stenoses; and the remaining patients had esophageal atresia. In 17 cases an isoperistaltic gastric tube was brought up to the neck through the retrosternal space; in 2 an anisoperistaltic gastric tube was used, cervical mobilization being via the posterior mediastinum without a thoracotomy in 1 case and by the left transpleural route in the other. The cervical anastomosis was carried out in one surgical stage in 12 patients and in two stages in 7. In 1 patient the tube was resected due to necrosis of its proximal third; the child later underwent an esophagocoloplasty. Necrosis of the colic graft, mediastinitis, and septicemia occurred, leading to the only death in the series. There were 12 fistulas of the cervical anastomosis (63.3%) and 8 stenoses (42.1%). All fistulas, with 1 exception, closed spontaneously after 8 days to 2 months, and all stenoses were treated by endoscopic dilatation. Another patient developed a fistula of the gastric tube with chronic evolution to a stenosis of the distal third of the tube and communication with the right lower pulmonary lobe. A lobectomy and closure of the fistula were necessary. All patients were followed for a period of 1 to 16 years. At present, all of them swallow solid food normally. The evolution of the nutritional status was normal (eutrophic) in 14 of the 18 patients (77.7%) who survived the operation; 4 showed variable degrees of malnutrition. In 2 of these 4 cases the malnutrition was due to poor socioeconomic conditions, but was not related to the surgery. Redundancy, a problem associated with esophagocoloplasty, was not observed in any of the gastric tubes, which was attributed to the thickness of the gastric wall. The authors prefer the use of an isoperistaltic gastric tube (with proximal base) for esophageal replacement in children and recommend that the operation should be carried out when the child is able to swallow solid foods and walk. As in any other major surgical procedure, a good nutritional state is essential prior to operation.


Halving of mortality at 1 year by domiciliary thrombolysis in the Grampian Region Early Anistreplase Trial (GREAT).
Our aim was to determine the time saved by administration of thrombolytic therapy at home rather than in the hospital and to assess whether earlier thrombolysis resulted in decreased mortality from acute myocardial infarction. There is much theoretic, experimental and trial evidence to indicate that in acute myocardial infarction the earlier that thrombolytic therapy is given, the greater its efficacy. However, the clinical importance of this time effect is uncertain. In a randomized double-blind parallel-group clinical trial, 311 patients with suspected acute myocardial infarction seen by their general practitioners within 4 h of symptom onset were given intravenous anistreplase (30 U) either at home or later, after arrival in the hospital. Anistreplase was given at home or in the hospital at median times of 101 and 240 min, respectively, after symptom onset. The median time saved by domiciliary thrombolysis was 130 min. By the end of 1 year after trial entry, 17 (10.4%) of 163 patients given anistreplase at home died compared with 32 (21.6%) of 148 in those allotted anistreplase in the hospital (relative reduction 52%, 95% confidence interval 14% to 89%, p = 0.007). In this trial the time saved by domiciliary thrombolysis by primary care physicians was > 2 h. It is likely that a similar time saving would be achieved if prehospital thrombolysis were to become established practice. Prehospital thrombolysis resulted in a halving of the mortality rate from acute myocardial infarction.


A Brief History of Promoter Development for Use in Transgenic Maize Applications.
Promoters regulate gene expression, and are essential biotechnology tools. Since its introduction in the mid-1990s, biotechnology has greatly enhanced maize productivity primarily through the development of insect control and herbicide tolerance traits. Additional biotechnology applications include improving seed nutrient composition, industrial protein production, therapeutic production, disease resistance, abiotic stress resistance, and yield enhancement. Biotechnology has also greatly expanded basic research into important mechanisms that govern plant growth and reproduction. Many novel promoters have been developed to facilitate this work, but only a few are widely used. Transgene optimization includes a variety of strategies some of which effect promoter structure. Recent reviews examine the state of the art with respect to transgene design for biotechnology applications. This chapter examines the use of transgene technology in maize, focusing on the way promoters are selected and used. The impact of new developments in genomic technology on promoter structure is also discussed.


[Specificity of the enzymes of Salmonella O-antigen biosynthesis. 6. Synthesis of sugar nucleotides with glycosyl phosphate activation. Analogs of guanosine diphosphate mannose modified at position 6 ofthe purine ring].
Interaction of alpha-D-mannopyranosyl phosphate with diphenyl phosphochloridate gave the trisubstituted pyrophosphate which was converted through the reaction with nucleoside 5'-phosphates into nucleoside 5'-(alpha-D-mannopyranosyl)pyrophosphates. The method was used for preparation of guanosine diphosphate mannose analogs derived from adenine, purine, 2-aminopurine, 2-amino-6-methoxypurine, 2-amino-6-chloropurine, and 2-amino-6-mercaptopurine. These analogs are necessary for study on substrate specificity of mannosyltransferases of Salmonella O-specific polysaccharides biosynthesis.


Children's use of anatomically detailed dolls to report genital touching in a medical examination: developmental and gender comparisons.
The influence of anatomically detailed dolls on 3- and 4-year-old children's reports of a routine medical exam was examined. During the exam, half the children received a genital examination and half the children did not. Immediately after the exam, all children were asked to demonstrate various events on an anatomically detailed doll and on their own bodies. Although there were age differences in the number of accurate demonstrations of how the doctor used different instruments, there were no age differences on measures that assessed reports of genital touching. Among girls, commission errors were more frequent when dolls were used because some children falsely showed insertions into the anal or genital cavity. The authors conclude that dolls should not be used in interviews with children below the age of 5 years.


Improved dewatering behavior of clay minerals dispersions via interfacial chemistry and particle interactions optimization.
Orthokinetic flocculation of clay dispersions at pH 7.5 and 22 degrees C has been investigated to determine the influence of interfacial chemistry and shear on dewatering and particle interactions behavior. Modification of pulp chemistry and behavior was achieved by using kaolinite and Na-exchanged (swelling) smectite clay minerals, divalent metal ions (Ca(II), Mn(II)) as coagulants and anionic polyacrylamide copolymer (PAM A) and non-ionic polyacrylamide homopolymer (PAM N) as flocculants. The pivotal role of shear, provided by a two-blade paddle impeller, was probed as a function of agitation rate (100-500 rpm) and time (15/60 s). Particle zeta potential and adsorption isotherms were measured to quantify the interfacial chemistry, whilst rheology and cryogenic SEM were used to investigate particle interactions and floc structure and aggregate network, respectively. Osmotic swelling, accompanied by the formation of "honeycomb" particle network structure and high yield stress, was produced by the Na-exchanged smectite, but not kaolinite, dispersions. Dispersion of the clay particles in 0.05 M Ca(II) or Mn(II) solution led to a marked reduction in particle zeta potential, complete suppression of swelling, honeycomb network structure collapse and a concomitant reduction in shear yield stress of smectite pulps. Optimum conditions for improved, orthokinetic flocculation performance of negatively charged clay particles, reflecting faster settling flocs comprised (i) coagulation, (ii) moderate agitation rate, (iii) shorter agitation time, and (iv) anionic rather than non-ionic PAM. The optimum dewatering rates were significantly higher than those produced by standard, manual-mixing flocculation techniques (plunging and cylinder inversion) commonly used in industry for flocculant trials. The optimum flocculation conditions did not, however, have a significant impact on the final sediment solid content of 20-22 wt%. Further application of shear to pre-sedimented pulps improved consolidation by 5-7 wt% solid. Higher shear yield stresses and greater settling rates were displayed by PAM A based than PAM N based pulps and this is attributed to the former's more expanded interfacial conformation and greater clay particles bridging ability. It appears that the intrinsic clay particles' physico-chemical properties and interactions limit compact pulp consolidation.


Advances in cell and gene-based therapies for cystic fibrosis lung disease.
Cystic fibrosis (CF) is a disease characterized by airway infection, inflammation, remodeling, and obstruction that gradually destroy the lungs. Direct delivery of the cystic fibrosis transmembrane conductance regulator (CFTR) gene to airway epithelia may offer advantages, as the tissue is accessible for topical delivery of vectors. Yet, physical and host immune barriers in the lung present challenges for successful gene transfer to the respiratory tract. Advances in gene transfer approaches, tissue engineering, and novel animal models are generating excitement within the CF research field. This review discusses current challenges and advancements in viral and nonviral vectors, cell-based therapies, and CF animal models.


Long-Term Association Between Disease Activity and Disability in Early Axial Spondyloarthritis: Results From a Prospective Observational Study of Inflammatory Back Pain.
Our primary objective was to study the long-term association between disease activity and disability in axial spondyloarthritis (SpA). Our secondary objective was to define patient profiles according to their level of disability. We analyzed data collected during the first 5 years of follow-up of a large early axial SpA cohort, the Devenir des Spondylarthropathies Indifferénciées Récentes (DESIR) cohort. Multivariable models were built to study the association between the Health Assessment Questionnaire for Ankylosing Spondylitis (HAQ-AS) and the Ankylosing Spondylitis Disease Activity Score with C-reactive protein (ASDAS-CRP), adjusting for potential confounders. Hierarchical multivariable analysis was conducted using the chi-square automatic interaction detector (CHAID) method, to help determine how variables best cluster to explain HAQ-AS. Data from 644 patients and 5,152 visits were analyzed. HAQ-AS was longitudinally, independently, and positively associated with ASDAS-CRP (adjusted B [adjB] 0.205 [95% confidence interval (95% CI) 0.187, 0.222]), the enthesitis score (adjB 0.011 [95% CI 0.008, 0.015]), the Bath Ankylosing Spondylitis Metrology Index (adjB 0.087 [95% CI 0.069, 0.105]), and female sex (adjB 0.172 [95% CI 0.120, 0.225]). The CHAID decision tree revealed ASDAS-CRP as the first variable with discriminative power on HAQ-AS. The cutoffs that separated different patient disability profiles were obtained. Disease activity contributes longitudinally to disability and is hierarchically superior to any other variable in explaining this health domain. Enthesitis and spinal mobility are also key drivers of disability in early axial SpA. ASDAS-CRP cutoffs that separated different patient disability profiles largely mimicked the cutoffs previously defined for ASDAS-CRP disease activity states.


Fetal echocardiography: where are we?
This article reviews the indications, technique and benefits of fetal echocardiography. The impact of fetal echocardiography has become evident with better surgical outcome of infants with prenatal diagnosis of congenital heart disease. A major use of this technique has also been shown in the field of fetal arrhythmias. Fetal echocardiography is not only used to diagnose a specific type of arrhythmia, it is also very helpful in assessing the effect of the arrhythmia on the fetus and in guiding transplacental therapy. However it is important to remember that even experienced echocardiographers can make both false positive and false negative diagnosis of congenital heart disease. As fetal cardiac interventions including fetal cardiac surgery loom on the horizon, fetal echocardiography assumes further importance.


Insights from system dynamics applications in addiction research: A scoping review.
Substance misuse and use disorders are dynamic and complex problems, situated within systems of interacting social, environmental, and neurobiological factors. System dynamics (SD) methods broaden, test, and improve understanding of complex systems and can help inform effective action. We sought to systematically review the use of SD tools in addiction-related research. Following PRISMA guidelines, we searched several databases from 1958 to 2019. We included studies focused on addiction-related screening and diagnosis, treatment, and return to use, as well as studies focused on earlier stages that may begin a path to addiction (e.g., experimentation, misuse onset). We extracted information from 59 articles with a median publication year of 2014. In addition to using SD to understand the underlying complexity driving addiction-related trends, other commonly cited reasons for use of SD included assessing impacts of potential actions (n = 35), predicting future trends (n = 28), and supporting strategic planning processes (n = 22). Most studies included simulation models (n = 43); however, some presented insights from qualitative SD diagrams (n = 9) and concept models (n = 6). The majority of studies focused on stages leading to potential addiction: initiation/ experimentation (n = 42) and misuse onset (n = 38). One-third (n = 20) engaged persons with lived experience or other stakeholders during the modeling process. Addiction-related SD research has increased over the last few decades with applications varying in several ways, from model purpose and types of data used to stakeholder involvement. Future applications should consider the benefits of stakeholder engagement throughout the modeling process and expanding models to include concomitant substance use.


Role of Surgery in Metastatic Renal Cell Carcinoma.
Surgery has historically been the mainstay of treatment for metastatic renal cell carcinoma (mRCC), but recent clinical trials demonstrated that contemporary systemic therapies alone are non-inferior to cytoreductive nephrectomy (CN). Thus, the current role of surgery is not precisely defined. CN remains an appropriate upfront treatment for the palliation of severe symptoms, select cases of metastatic non-clear cell renal cell carcinoma, for consolidation following systemic therapy, and in the setting of oligometastatic disease. Metastasectomy is ideally utilized to achieve a disease-free state when there is minimal morbidity associated with surgery. Given the heterogenous nature of mRCC, the decision for systemic therapy and surgery should be made through a multidisciplinary approach tailored to each individual patient.


Different distribution of plexiform lesions in primary and secondary pulmonary hypertension.
Despite much interest in plexiform lesions, no published work compares their distribution in different types of pulmonary hypertension. Scattered reports of plexiform lesions in bronchial arteries oppose the consensus view that the lesions develop in pulmonary arteries. To compare the localization of plexiform lesions in different types of pulmonary hypertension, and to assess the role of the bronchial arteries in their formation, we examined by light microscopy lung tissue from five patients with primary plexogenic pulmonary arteriopathy (PPPA), six with pulmonary hypertension secondary to congenital cardiac malformations (CCM), and one with pulmonary hypertension complicating hepatic cirrhosis. We classified the 270 plexiform lesions observed as either preacinar or intra-acinar based on the type of pulmonary artery in which they were located, and computed the frequencies of each type of lesion within each etiologic group. We searched for lesions developing in bronchial arteries. Then, postulating that a close anatomic relationship between plexiform lesions and bronchial arteries would necessitate a clustering of the lesions near sites in the lung subserved by the bronchial circulation, we measured, for 211 of the 270 lesions previously classified, the distance from the lesion to the nearest airway and computed the mean lesion-to-airway distance in each etiologic group. The frequencies of preacinar plexiform lesions were 34% in PPPA, 67% in CCM (P < .01), and 21% in the case of cirrhosis. We found no plexiform lesions within bronchial arteries, and the mean plexiform lesion-to-airway distances were 1,680 +/- 180 microns in PPPA, 1,330 +/- 220 microns in CCM, and 2,050 +/- 1,090 microns in cirrhosis (P > .05). Our data suggest that (1) the distribution of plexiform lesions within the pulmonary arterial tree varies depending on the etiology, (2) plexiform lesions rarely if ever arise in bronchial arteries, and (3) plexiform lesions are not preferentially distributed near parts of the lung subserved by the bronchial circulation.


Driving patterns before and after hospitalization for heart disease.
Persons hospitalized with heart disease were licensed as often in the pre-hospital period and drove slightly (but not significantly) fewer miles than did healthier persons of similar age, sex and neighborhood. After discharge, those who returned to driving reduced their mileage and drove under different conditions. This occurred both because of overall changes in life style (e.g. retirement) associated with illness and because of health effects on driving per se. In fact, while activities changed, the methods of transportation used to accomplish these remained the same, with driving oneself being the preferred mode. Physicians advised patients only about driving during the immediate recovery period rather than with an eye toward long-term safety risk. Few people reported their heart condition to the Department of Motor Vehicles. A functional severity score based on primary and comorbid conditions was developed to distinguish individuals with different amounts of impairment relevent to driving.


[Knowledge, attitude and opinion of patients regarding the new German legislation on advance care planning : Results of a survey in a department of general internal medicine].
In September 2009 a new legislation for advance care planning was introduced in Germany with the important characteristics of bindingness and unlimited validity for individual directives. Knowledge regarding this act and the attitude towards its characteristics among patients is unclear. Analysis of knowledge, attitude and opinion of patients in a general internal medical department regarding advance care planning in general and the recent German legislation. A total of 200 consecutive patients in an internal medicine ward were interviewed with the help of a questionnaire regarding their attitude to and knowledge on advance care planning in general and the current legislation. Approximately 40 % of the patients had issued some form of directive (either advance care directive or health care proxy) and only 7.5 % were advised by their physicians to make an advance directive. Patients with no directive were not willing to deal with dying and death, were not well-informed about directives or assumed that relatives or physicians would make an appropriate decision. Characteristics of the new legislation were controversially assessed; only 21 % of the patients wished to have a literal implementation of their directive. Regarding the content of an advance directive, more than 80 % of the patients voted for pain control in the palliative setting. The proportion of patients with a directive regarding advance care planning is only slowly increasing. Many patients are not well-informed, do not want to deal with dying or would like to delegate decisions to relatives and physicians. The present characteristics of the German legislation are controversially assessed and often do not represent the wishes of the patients.


Mab labelled liposomes as selective vehicles for drugs in H.I.V. infected cells.
Being able to carry zidovudine (AZT) at known concentrations into CD4+/CD38+ and CD14+ cells permits: - to reduce the drug dosage and to increase the interval for administration (until 1 dose I.V. every week); - to modulate the drug concentration into the CD4+/CD38 an CD14+ cells in relation to the "in vitro" determined HIV sensitiveness; - to eliminate haematological, medullary and general toxicity; - to be able to treat severely hill patients. Further studies are necessary in order to: - To find out the better phase to start the therapy; - To use several drugs with different mechanisms of action in order to slow down as much as possible the presence of resistant viral strains. - As for other drugs which are beginning to be used with artificial vehicles, futher studies are required to improve the selectivity and safety of LIPOAZT for the target cells including macrophages.


K1K8: an Hp1404-derived antibacterial peptide.
As an alternative class of antimicrobial agents used to overcome drug-resistant infections, antimicrobial peptides (AMPs) have recently gained significant attention. In this study, we designed an improved antimicrobial peptide, K1K8, based on the molecular template of Hp1404. Compared to the wild-type Hp1404, K1K8 showed an improved antibacterial spectrum in vitro, a lower hemolytic activity, and an enhanced serum stability. Importantly, K1K8 also decreased methicillin-resistant Staphylococcus aureus (MRSA) bacterial counts in the wounded region in a mouse skin infection model. Interestingly, K1K8 did not induce bacterial resistance or non-specific immune response reactions. Moreover, the peptide killed bacterial cells mainly by disrupting the bacterial membrane. In summary, K1K8 has the potential to be used as an improved anti-infection agent for topical use, which opens an avenue that potential anti-infection drugs may be designed and developed from the molecular templates of AMPs.


Molecular epidemiological study of clinical Acinetobacter baumannii isolates: phenotype switching of antibiotic resistance.
The presence of clinical Acinetobacter baumannii (A. baumannii) isolates with differing antibiotic resistance phenotypes in the same patient causes difficulties and confusion in treatment. This phenomenon may be caused by reasons such as cross-infection from neighboring patients that switches to different A. baumannii strain, natural mutation of A. baumannii, inducing of different antibiotic resistance genes expression or acquisition of genes conferring resistance from another source. To elucidate this question, clinical A. baumannii strains, isolated from the same individual patients, showed antibiotic resistance phenotypes switching during the same hospitalization period, were attentively collected for further analysis. Molecular approaches for phylogenetic analysis, including pulsed field gel electrophoresis, multilocus sequence typing, and short tandem repeat analysis, were employed for the chronological studies. Our results showed that antibiotic resistance phenotype switching could have occurred as a result through both cross-infection and natural mutation roots. Our results also suggest that rapid phenotype switching between paired isolates could occur during one single course of antibiotic treatment. Though cross infection caused antibiotic resistance phenotype switching does occur, natural mutation of A. baumannii isolates is particularly cautious for antibiotic treatment.


High levels of HMG1-2 protein expression in the cytoplasm and nucleus of hydrocortisone sensitive amphibian thymocytes.
A major 26 kDa protein was identified in the cytoplasmic and nuclear compartments of axolotl thymocytes. A polyclonal antiserum was produced against the denatured form of this protein. High levels of 26 kDa were expressed by hydrocortisone-sensitive lymphocytes which represent a major thymocyte subpopulation in young animals. However, no further expression of the 26 kDa protein was observed in involuted thymus of adult animals nor in thymus of young artificially metamorphosed axolotls. The 26 kDa was never expressed by splenic and blood peripheral lymphocytes at any stage of development. Partial N-terminal amino acid sequence and amino acid composition demonstrate that the 26 kDa polypeptide is strongly homologous to HMG1-2 proteins, the most abundant members of the high mobility group (HMG) non-histone chromosomal proteins. HMG1-2 are thought to be involved in the organization of chromatin structure, as well as in the stability, replication and transcription of DNA. It was confirmed that the 26 kDa axolotl polypeptide is recognized by a well characterized rabbit antiserum to rat HMG1-2 proteins.


Time to workforce exit after a Parkinson's disease diagnosis.
The impact of Parkinson's disease (PD) on workforce participation has received little attention even though demographic, lifestyle, and political changes together will result in an increased burden of PD on the working-age population. In this study, we investigate workforce survival after a PD diagnosis, as well as what demographic factors that are associated with workforce survival. As an exploratory outcome, we investigate workforce survival in persons with and without device-aided treatment (DAT). This is a nested case-cohort study based on Swedish national data from 2001-2016. Controls were matched on year of birth, sex, and municipality of residence. The used registers contain data on demographics, social insurance, in- and outpatient visits, filled drug prescriptions, and cause of death on the person-level. A total of 4781 persons with PD and 23,905 controls were included. The median survival until all-cause workforce exit was 43 months among persons that were workforce-active at the time of PD diagnosis, compared to 66 months in non-PD controls. Being female, ≥50 years old at diagnosis, or having a lower education were contributing factors to health-related workforce exit. Persons receiving DAT during follow-up exhibited shorter workforce survival than controls. However, this needs further investigation, particularly as patients have generally already left the workforce at the time for start of DAT. It is evident that PD has grave negative effects on workforce participation. Thus, supportive measures need to start at an early stage after diagnosis, and the development of new interventions is urgently needed.


Tissue biomarkers for diagnosis & management of oral squamous cell carcinoma.
Squamous cell carcinoma (SCC) accounts for more than 90% of malignancies of the oral cavity and oropharynx. Globally, SCC is one of the top ten cancers with a predilection for older males. In the U.S., SCC accounts for more deaths annually than cervical cancer, malignant melanoma or Hodgkin's lymphoma and costs about 2 billion dollars in treatment expenses. The 5-year survival rate is less than 50%, a prognosis that is poorer than that of breast cancer or melanoma.2 Significant numbers of patients develop local recurrence, second primary tumors and distant metastases. Patients presenting with late stage disease resulting in poor survival, are common; in part due to inadequate screening protocols and access to care issues. In the following review article, the risk factors, current screening protocols, treatment options and biomarkers for diagnosis and prognosis of SCC, will be discussed.