bluuebunny/crossref_metadata_2025 huggingface dataset:
Lactococci and lactobacilli as mucosal delivery vectors for therapeutic proteins and DNA vaccines
Authors: Bermúdez-Humarán Luis G , Kharrat Pascale , Chatel Jean-Marc , Langella Philippe
Published: August 2011

Food-grade Lactic Acid Bacteria (LAB) have been safely consumed for centuries by humans in fermented foods. Thus, they are good candidates to develop novel oral vectors, constituting attractive alternatives to attenuated pathogens, for mucosal delivery strategies. Herein, this review summarizes our research, up until now, on the use of LAB as mucosal delivery vectors for therapeutic proteins and DNA vaccines. Most of our work has been based on the model LAB Lactococcus lactis, for which we have developed efficient genetic tools, including expression signals and host strains, for the heterologous expression of therapeutic proteins such as antigens, cytokines and enzymes. Resulting recombinant lactococci strains have been tested successfully for their prophylactic and therapeutic effects in different animal models: i) against human papillomavirus type 16 (HPV-16)-induced tumors in mice, ii) to partially prevent a bovine β-lactoglobulin (BLG)-allergic reaction in mice and iii) to regulate body weight and food consumption in obese mice. Strikingly, all of these tools have been successfully transposed to the Lactobacillus genus, in recent years, within our laboratory. Notably, anti-oxidative Lactobacillus casei strains were constructed and tested in two chemically-induced colitis models. In parallel, we also developed a strategy based on the use of L. lactis to deliver DNA at the mucosal level, and were able to show that L. lactis is able to modulate the host response through DNA delivery. Today, we consider that all of our consistent data, together with those obtained by other groups, demonstrate and reinforce the interest of using LAB, particularly lactococci and lactobacilli strains, to develop novel therapeutic protein mucosal delivery vectors which should be tested now in human clinical trials.
Genomic analysis reveals Lactobacillus sanfranciscensis as stable element in traditional sourdoughs
Authors: Vogel Rudi F , Pavlovic Melanie , Ehrmann Matthias A , Wiezer Arnim , Liesegang Heiko , Offschanka Stefanie , Voget Sonja , Angelov Angel , Böcker Georg , Liebl Wolfgang
Published: August 2011

Sourdough has played a significant role in human nutrition and culture for thousands of years and is still of eminent importance for human diet and the bakery industry.Lactobacillus sanfranciscensisis the predominant key bacterium in traditionally fermented sourdoughs.

The genome ofL. sanfranciscensisTMW 1.1304 isolated from an industrial sourdough fermentation was sequenced with a combined Sanger/454-pyrosequencing approach followed by gap closing by walking on fosmids. The sequencing data revealed a circular chromosomal sequence of 1,298,316 bp and two additional plasmids, pLS1 and pLS2, with sizes of 58,739 bp and 18,715 bp, which are predicted to encode 1,437, 63 and 19 orfs, respectively. The overall GC content of the chromosome is 34.71%. Several specific features appear to contribute to the ability ofL. sanfranciscensisto outcompete other bacteria in the fermentation.L. sanfranciscensiscontains the smallest genome within the lactobacilli and the highest density of ribosomal RNA operons per Mbp genome among all known genomes of free-living bacteria, which is important for the rapid growth characteristics of the organism. A high frequency of gene inactivation and elimination indicates a process of reductive evolution. The biosynthetic capacity for amino acids scarcely availably in cereals and exopolysaccharides reveal the molecular basis for an autochtonous sourdough organism with potential for further exploitation in functional foods. The presence of two CRISPR/cas lociversusa high number of transposable elements suggests recalcitrance to gene intrusion and high intrinsic genome plasticity.
Inhibitory activity of Lactobacillus plantarum LMG P-26358 against Listeria innocua when used as an adjunct starter in the manufacture of cheese
Authors: Mills Susan , Serrano LMariela , Griffin Carmel , O'Connor Paula M , Schaad Gwenda , Bruining Chris , Hill Colin , Ross RPaul , Meijer Wilco C
Published: August 2011

Lactobacillus plantarum LMG P-26358 isolated from a soft French artisanal cheese produces a potent class IIa bacteriocin with 100% homology to plantaricin 423 and bacteriocidal activity against Listeria innocua and Listeria monocytogenes. The bacteriocin was found to be highly stable at temperatures as high as 100°C and pH ranges from 1-10. While this relatively narrow spectrum bacteriocin also exhibited antimicrobial activity against species of enterococci, it did not inhibit dairy starters including lactococci and lactobacilli when tested by well diffusion assay (WDA). In order to test the suitability of Lb. plantarum LMG P-26358 as an anti-listerial adjunct with nisin-producing lactococci, laboratory-scale cheeses were manufactured. Results indicated that combining Lb. plantarum LMG P-26358 (at 108 colony forming units (cfu)/ml) with a nisin producer is an effective strategy to eliminate the biological indicator strain, L. innocua. Moreover, industrial-scale cheeses also demonstrated that Lb. plantarum LMG P-26358 was much more effective than the nisin producer alone for protection against the indicator. MALDI-TOF mass spectrometry confirmed the presence of plantaricin 423 and nisin in the appropriate cheeses over an 18 week ripening period. A spray-dried fermentate of Lb. plantarum LMG P-26358 also demonstrated potent anti-listerial activity in vitro using L. innocua. Overall, the results suggest that Lb. plantarum LMG P-26358 is a suitable adjunct for use with nisin-producing cultures to improve the safety and quality of dairy products.
Enchanced levels of apolipoprotein M during HBV infection feedback suppresses HBV replication
Authors: Gu Jin-Gang , Zhu Cheng-liang , Cheng Duo-zhi , Xie Yan , Liu Fang , Zhou Xin
Published: August 2011

Background

Chronic liver diseases can interfere with hepatic metabolism of lipoproteins, apolipoproteins. Hepatitis B virus (HBV) is a major etiological agent causing acute and chronic liver diseases. Apolipoprotein M (ApoM) is a high-density lipoprotein (HDL) apolipoprotein and exclusively expressed in the liver parenchyma cells and in the tubular cells of the kidney. This study was to determine the correlation between HBV infection and ApoM expression.

Materials and methods

Serum ApoM levels in patients with HBV infection and in healthy individuals were measured by ELISA, ApoM mRNA expression were determined by RT-PCR, and the expression of S and E proteins of HBV, as well as the synthesis of viral DNA were measured by ELISA and real-time PCR.

Results

The levels of serum ApoM was significantly elevated in patients as compared to healthy individuals (P< 0.001), ApoM promoter activity, mRNA and protein expression were all stimulated in cells transfected with infectious HBV clone. In addition, ApoM decreases the expression of S and E proteins of HBV and the synthesis of viral DNA.

Conclusion

Raised ApoM levels in HBV infection may in turn suppress HBV replication, one of the protective mechanisms of nature.
Activation of monocytes and cytokine production in patients with peripheral atherosclerosis obliterans
Authors: Corrêa Camila R , Dias-Melicio Luciane A , Calvi Sueli A , Lastória Sidney , Soares Angela MVC
Published: August 2011

Background

Arterial peripheral disease is a condition caused by the blocked blood flow resulting from arterial cholesterol deposits within the arms, legs and aorta. Studies have shown that macrophages in atherosclerotic plaque are highly activated, which makes these cells important antigen-presenting cells that develop a specific immune response, in which LDLox is the inducing antigen. As functional changes of cells which participate in the atherogenesis process may occur in the peripheral blood, the objectives of the present study were to evaluate plasma levels of anti-inflammatory and inflammatory cytokines including TNF-α, IFN-γ, interleukin-6 (IL-6), IL-10 and TGF-β in patients with peripheral arteriosclerosis obliterans, to assess the monocyte activation level in peripheral blood through the ability of these cells to release hydrogen peroxide (H2O2) and to develop fungicidal activity against Candida albicans (C. albicans) in vitro.

Methods

TNF-α, IFN-γ, IL-6, IL-10 and TGF-β from plasma of patients were detected by ELISA. Monocyte cultures activated in vitro with TNF-alpha and IFN-gamma were evaluated by fungicidal activity against C. albicans by culture plating and Colony Forming Unit (CFU) recovery, and by H2O2 production.

Results

Plasma levels of all cytokines were significantly higher in patients compared to those detected in control subjects. Control group monocytes did not release substantial levels of H2O2
in vitro, but these levels were significantly increased after activation with IFN-γ and TNF-α. Monocytes of patients, before and after activation, responded less than those of control subjects. Similar results were found when fungicidal activity was evaluated. The results seen in patients were always significantly smaller than among control subjects. Conclusions: The results revealed an unresponsiveness of patient monocytes in vitro probably due to the high activation process occurring in vivo as corroborated by high plasma cytokine levels.
Upper limb artery segmental occlusions due to chronic use of ergotamine combined with itraconazole, treated by thrombolysis
Authors: Cervi Edoardo , Bonardelli Stefano , Battaglia Giuseppe , Gheza Federico , Maffeis Roberto , Nodari Franco , Maroldi Roberto , Giulini Stefano M
Published: August 2011

Background

The ergotamine tartrate associated with certain categories of drugs can lead to critical ischemia of the extremities. Discontinuation of taking ergotamine is usually sufficient for the total regression of ischemia, but in some cases it could be necessary thrombolytic and anticoagulant therapy to avoid amputation.

Case report

A woman of 62 years presented with a severe pain left forearm appeared 10 days ago, with a worsening trend. The same symptoms appeared after 5 days also in the right forearm. Physical examination showed the right arm slightly hypothermic, with radial reduced pulse in presence of reduced sensitivity. The left arm was frankly hypothermic, pulse less on radial and with an ulnar humeral reduced pulse, associated to a decreased sensitivity and motility.

Clinical history shows a chronic headache for which the patient took a daily basis for years Cafergot suppository (equivalent to 3.2 mg of ergotamine).

From about ten days had begun therapy with itraconazole for vaginal candidiasis. The Color-Doppler ultrasound shown arterial thrombosis of the upper limbs (humeral and radial bilateral), with minimal residual flow to the right and no signal on the humeral and radial left artery.

Results

Angiography revealed progressive reduction in size of the axillary artery and right humeral artery stenosis with right segmental occlusions and multiple hypertrophic collateral circulations at the elbow joint. At the level of the right forearm was recognizable only the radial artery, decreased in size. Does not recognize the ulnar, interosseous artery was thin. To the left showed progressive reduction in size of the distal subclavian and humeral artery, determined by multiple segmental steno-occlusion with collateral vessels serving only a thin hypotrophic interosseous artery.

Arteriographic findings were compatible with systemic drug-induced disease. The immediate implementation of thrombolysis, continued for 26 hours, with heparin in continuous intravenous infusion and subsequent anticoagulant therapy allowed the gradual disappearance of the symptoms with the reappearance of peripheral pulses.

Conclusion

Angiography showed regression of vasospasm and the resumption of flow in distal vessels. The patient had regained sensitivity and motility in the upper limbs and bilaterally radial and ulnar were present.
Tissue inhibitor of metalloproteinases-3 mediates the death of immature oligodendrocytes via TNF-α/TACE in focal cerebral ischemia in mice
Authors: Yang Yi , Jalal Fakhreya Y , Thompson Jeffrey F , Walker Espen J , Candelario-Jalil Eduardo , Li Lu , Reichard Ross R , Ben Chi , Sang Qing-Xiang , Cunningham Lee Anna , Rosenberg Gary A
Published: August 2011

Background and Purpose

Oligodendrocyte (OL) death is important in focal cerebral ischemia. TIMP-3 promotes apoptosis in ischemic neurons by inhibiting proteolysis of TNF-α superfamily of death receptors. Since OLs undergo apoptosis during ischemia, we hypothesized that TIMP-3 contributes to OL death.

Methods

Middle cerebral artery occlusion (MCAO) was induced inTimp-3knockout (KO) and wild type (WT) mice with 24 or 72 h of reperfusion. Cell death in white matter was investigated by stereology and TUNEL. Mature or immature OLs were identified using antibodies against glutathioneS-transferase-π (GST-π) and galactocerebroside (GalC), respectively. Expression and level of proteins were examined using immunohistochemistry and immunoblotting. Protein activities were determined using a FRET peptide.

Results

Loss of OL-like cells was detected at 72 h only in WT ischemic white matter where TUNEL showed greater cell death. TIMP-3 expression was increased in WT reactive astrocytes. GST-π was reduced in ischemic white matter of WT mice compared with WT shams with no difference between KO and WT at 72 h. GalC level was significantly increased in both KO and WT ischemic white matter at 72 h. However, the increase in GalC in KO mice was significantly higher than WT; most TUNEL-positive cells in ischemic white matter expressed GalC, suggesting TIMP-3 deficiency protects the immature OLs from apoptosis. There were significantly higher levels of cleaved caspase-3 at 72 h in WT white matter than in KO. Greater expression of MMP-3 and -9 was seen in reactive astrocytes and/or microglia/macrophages in WT at 72 h. We found more microglia/macrophages in WT than in KO, which were the predominant source of increased TNF-α detected in the ischemic white matter. TACE activity was significantly increased in ischemic WT white matter, which was expressed in active microglia/macrophages and OLs.

Conclusions

Our results suggested that focal ischemia leads to proliferation of immature OLs in white matter and that TIMP-3 contributes to a caspase-3-dependent immature OL death via TNF-α-mediated neuroinflammation. Future studies will be needed to delineate the role of MMP-3 and MMP-9 that were increased in theTimp-3wild type.
Does human bocavirus infection depend on helper viruses? A challenging case report
Authors: Streiter Monika , Malecki Monika , Prokop Aram , Schildgen Verena , Lüsebrink Jessica , Guggemos Andreas , Wißkirchen Matthias , Weiß Michael , Cremer Reinhold , Brockmann Michael , Schildgen Oliver
Published: August 2011

A case of severe diarrhoea associated with synergistic human bocavirus type 1 (HBoV) and human herpes virus type 6 (HHV6) is reported. The case supports the hypotheses that HBoV infection under clinical conditions may depend on helper viruses, or that HBoV replicates by a mechanism that is atypical for parvoviruses, or that HBoV infection can be specifically treated with cidofovir.
Infective endocarditis caused by methicillin-resistant Staphylococcus aureus in a young woman after ear piercing: a case report
Authors: Nah So-Yun , Chung Moon-Hyun , Park Jae Eun , Durey Areum , Kim Mijeong , Lee Jin-Soo
Published: August 2011

Introduction

Ear piercing is a common practice among Korean adolescents and young women and usually is performed by nonmedical personnel, sometimes under suboptimal hygienic conditions. Consequently, ear piercing has been associated with various infectious complications, including fatal infective endocarditis. We report a case of infective endocarditis that was caused by community-associated methicillin-resistant Staphylococcus aureus after ear piercing and that was accompanied by a noticeable facial rash.

Case presentation

A 29-year-old Korean woman underwent ear piercing six days before hospitalization. On admission, she had fever, erythematous maculopapular rashes on her face, signs of generalized emboli, vegetation in her mitral valve, and methicillin-resistant S. aureus bacteremia. On the basis of the blood culture results, she was treated with vancomycin in combination with gentamicin. On day six of hospitalization, a rupture of the papillary muscle of her mitral valve developed, and emergency cardiac surgery replacing her mitral valve with a prosthetic valve was performed. After eight weeks of antibiotic therapy, she was treated successfully and discharged without significant sequelae.

Conclusions

Numerable cases of body piercing-related infective endocarditis have been reported, and since ear piercing is commonplace nowadays, the importance of risk recognition cannot be overemphasized. In our report, a patient developed infective endocarditis that was caused by methicillin-resistant S. aureus after ear piercing and that was accompanied by an interesting feature, namely facial rash.
Leishmania infantum HSP70-II null mutant as candidate vaccine against leishmaniasis: a preliminary evaluation
Authors: Carrión Javier , Folgueira Cristina , Soto Manuel , Fresno Manuel , Requena Jose M
Published: August 2011

Background

Visceral leishmaniasis is the most severe form of leishmaniasis and no effective vaccine exists. The use of live attenuated vaccines is emerging as a promising vaccination strategy.

Results

In this study, we tested the ability of aLeishmania infantumdeletion mutant, lacking bothHSP70-IIalleles (ΔHSP70-II), to provide protection againstLeishmaniainfection in theL. major-BALB/c infection model. Administration of the mutant line by either intraperitoneal, intravenous or subcutaneous route invariably leads to the production of high levels of NO and the development in mice of type 1 immune responses, as determined by analysis of anti-LeishmaniaIgG subclasses. In addition, we have shown that ΔHSP70-II would be a safe live vaccine as immunodeficient SCID mice, and hamsters (Mesocricetus auratus), infected with mutant parasites did not develop any sign of pathology.

Conclusions

The results suggest that the ΔHSP70-II mutant is a promising and safe vaccine, but further studies in more appropriate animal models (hamsters and dogs) are needed to appraise whether this attenuate mutant would be useful as vaccine against visceral leishmaniasis.
The short form endometriosis health profile (EHP-5): translation and validation study of the Iranian version
Authors: Goshtasebi Azita , Nematollahzadeh Maryam , Hariri Fatemeh Zahra , Montazeri Ali
Published: August 2011

Background

Endometriosis Health Profile (EHP-5) is a valid instrument to measure health-related quality of life in endometriosis. This study was conducted to culturally adapt and validate the EHP-5 in Iran.

Methods

Using a standard "forward-backward' translation procedure, the English language version of the questionnaire was translated into Persian (Iranian language). Then a sample of 199 women aged 18-50 years completed the questionnaire. To test reliability the internal consistency was assessed by Cronbach's alpha coefficient. Validity was evaluated using known groups comparison.

Results

The mean age of respondents was 31.4 (SD = 5.4) years. Reliability analysis showed satisfactory result (Cronbach's alpha coefficient = 0.71). The questionnaire discriminated well between sub-groups of women differing in infertility and premenstrual syndrome (PMS) in the expected direction.

Conclusion

This preliminary validation study of the Iranian version of the EHP-5 proved that it is an acceptable, reliable and valid measure of quality of life in endometriosis patients.
Local recurrence and assessment of sentinel lymph node biopsy in deep soft tissue leiomyosarcoma of the extremities
Authors: Lamyman Michael J , Giele Henk P , Critchley Paul , Whitwell Duncan , Gibbons Max , Athanasou Nicholas A
Published: August 2011

Background

Leiomyosarcoma of deep soft tissues of the extremities is a rare malignant tumour treated primarily by surgery. The incidence of local recurrence and lymph node metastasis is uncertain and it is not known whether a sentinel lymph node biopsy is indicated in these tumours.

Methods

A retrospective review of patients treated for extremity deep soft tissue leiomyosarcoma at our institution over a 10-year period was conducted. Patients developing local recurrence or lymph node metastasis were identified. The presence or absence of lymphatics in the primary tumours was assessed by immunohistochemical expression of LYVE-1 and podoplanin.

Results

27 patients (mean age 62 years) were included in the study. 15 were female and 12 male. Lymph node metastasis was seen in only two cases (7%); intratumoural lymphatics were identified in the primary tumours of both these cases. Local recurrence occurred in 25.9% of cases despite complete excision and post-operative radiotherapy; the mean time to recurrence was 10.1 months.

Conclusion

On the basis of this study, we do not advocate sentinel lymph node biopsy in this group of patients except in those cases in which intratumoural lymphatics can be demonstrated. Close follow up is important especially for high grade leiomyosarcomas, particularly in the first year, as these tumours have a high incidence of local recurrence.
Protein phosphatase 5 is a negative regulator of separase function during cortical granule exocytosis inC. elegans
Authors: Richie Christopher T. , Bembenek Joshua N. , Chestnut Barry , Furuta Tokiko , Schumacher Jill M. , Wallenfang Matthew , Golden Andy
Published: August 2011

Mutations in the Caenorhabditis elegans separase gene, sep-1, are embryonic lethal. Newly fertilized mutant embryos have defects in polar body extrusion, fail to undergo cortical granule exocytosis, and subsequently fail to complete cytokinesis. Chromosome nondisjunction during the meiotic divisions is readily apparent after depletion of sep-1 by RNAi treatment, but much less so in hypomorphic mutant embryos. To identify factors that influence the activity of separase in cortical granule exocytosis and cytokinesis, we carried out a genetic suppressor screen. A mutation in the protein phosphatase 5 (pph-5) gene was identified as an extragenic suppressor of sep-1. This mutation suppressed the phenotypes of hypomorphic separase mutants but not RNAi depleted animals. Depletion of pph-5 caused no phenotypes on its own, but was effective in restoring localization of mutant separase to vesicles and suppressing cortical granule exocytosis and cytokinesis phenotypes. The identification of PPH-5 as a suppressor of separase suggests that a new phospho-regulatory pathway plays an important role in regulating anaphase functions of separase.
Annexin B9 binds to βH-spectrin and is required for multivesicular body function in Drosophila
Authors: Tjota Monika , Lee Seung-Kyu , Wu Juan , Williams Janice A. , Khanna Mansi R. , Thomas Claire M.
Published: August 2011

The role of the cytoskeleton in protein trafficking is still being defined. Here, we describe a relationship between the small Ca2+-dependent membrane-binding protein Annexin B9 (AnxB9), apical βHeavy-spectrin (βH) and the multivesicular body (MVB) in Drosophila. AnxB9 binds to a subset of βH spliceoforms, and loss of AnxB9 results in an increase in basolateral βH and its appearance on cytoplasmic vesicles that overlap with the MVB markers Hrs, Vps16 and EPS15. Similar colocalizations are seen when βH-positive endosomes are generated either by upregulation of βH in pak mutants or through the expression of the dominant-negative version of βH. In common with other mutations disrupting the MVB, we also show that there is an accumulation of ubiquitylated proteins and elevated EGFR signaling in the absence of AnxB9 or βH. Loss of AnxB9 or βH function also causes the redistribution of the DE-Cadherin (encoded by shotgun) to endosomal vesicles, suggesting a rationale for the previously documented destabilization of the zonula adherens in karst (which encodes βH) mutants. Reduction of AnxB9 results in degradation of the apical–lateral boundary and the appearance of the basolateral proteins Coracle and Dlg on internal vesicles adjacent to βH. These results indicate that AnxB9 and βH are intimately involved in endosomal trafficking to the MVB and play a role in maintaining high-fidelity segregation of the apical and lateral domains.
Regulation of sororin by Cdk1-mediated phosphorylation
Authors: Dreier Megan R. , Bekier Michael E. , Taylor William R.
Published: August 2011

Tumor cells are commonly aneuploid, a condition contributing to cancer progression and drug resistance. Understanding how chromatids are linked and separated at the appropriate time will help uncover the basis of aneuploidy and will shed light on the behavior of tumor cells. Cohesion of sister chromatids is maintained by the multi-protein complex cohesin, consisting of Smc1, Smc3, Scc1 and Scc3. Sororin associates with the cohesin complex and regulates the segregation of sister chromatids. Sororin is phosphorylated in mitosis; however, the role of this modification is unclear. Here we show that mutation of potential cyclin-dependent kinase 1 (Cdk1) phosphorylation sites leaves sororin stranded on chromosomes and bound to cohesin throughout mitosis. Sororin can be precipitated from cell lysates with DNA–cellulose, and only the hypophosphorylated form of sororin shows this association. These results suggest that phosphorylation of sororin causes its release from chromatin in mitosis. Also, the hypophosphorylated form of sororin increases cohesion between sister chromatids, suggesting that phosphorylation of sororin by Cdk1 influences sister chromatid cohesion. Finally, phosphorylation-deficient sororin can alleviate the mitotic block that occurs upon knockdown of endogenous sororin. This mitotic block is abolished by ZM447439, an Aurora kinase inhibitor, suggesting that prematurely separated sister chromatids activate the spindle assembly checkpoint through an Aurora kinase-dependent pathway.
Disease-associated missense mutations in bestrophin-1 affect cellular trafficking and anion conductance
Authors: Milenkovic Vladimir M. , Röhrl Elena , Weber Bernhard H. F. , Strauss Olaf
Published: August 2011

Bestrophin-1, an integral membrane protein encoded by the BEST1 gene, is localized predominantly to the basolateral membrane of the retinal pigment epithelium. Mutations in the BEST1 gene have been associated with Best vitelliforme macular dystrophy (BMD), a central retinopathy with autosomal dominant inheritance and variable penetrance. Over 120 disease-causing mutations are known, the majority of which result in amino acid substitutions within four mutational hot-spot regions in the highly conserved N-terminal half of the protein. Although initially thought to impair Cl− channel function, the molecular pathology of BEST1 mutations is still controversial. We have analyzed the subcellular localization of 13 disease-associated BEST1 mutant proteins in polarized MDCK II cells, an established model of apical to basolateral protein sorting. Immunostaining demonstrated that nine of the 13 mutant proteins failed to integrate into the cell membrane. The defective proteins were predominantly retained in the cytoplasm, whereas wild-type bestrophin-1 revealed cell membrane localization. Functional analysis of I− fluxes in HEK-293 cells showed that all mutants exhibited a significant reduction in anion conductance. Our data indicate that defective intracellular trafficking could be a common cause of BMD accompanied by impaired anion conductance, representing a loss of anion channel function that is probably due to mistargeting of mutant protein.
Purification of the proprotein convertase furin by affinity chromatography based on PC-specific inhibitors
Authors: Kuester Miriam , Becker Gero L. , Hardes Kornelia , Lindberg Iris , Steinmetzer Torsten , Than Manuel E.
Published: August 2011

In eucaryotes, many secreted proteins and peptides are proteolytically excised from larger precursor proteins by a specific class of serine proteases, the proprotein/prohormone convertases (PCs). This cleavage is essential for substrate activation, making the PCs very interesting pharmacological targets in cancer and infectious disease research. Correspondingly, their structure, function and inhibition are intensely studied – studies that require the respective target proteins in large amounts and at high purity. Here we describe the development of a novel purification protocol of furin, the best-studied member of the PC family. We combined the heterologous expression of furin from CHO cells with a novel purification scheme employing an affinity step that efficiently extracts only active furin from the conditioned medium by using furin-specific inhibitor moieties as bait. Several potential affinity tags were synthesized and their binding to furin characterized. The best compound, Biotin-(Adoa)2-Arg-Pro-Arg-4-Amba coupled to streptavidin-Sepharose beads, was used in a three-step chromatographic protocol and routinely resulted in a high yield of a homogeneous furin preparation with a specific activity of ∼60 units/mg protein. This purification and the general strategy can easily be adapted to the efficient purification of other PC family members.
Oxidative folding: recent developments
Authors: Szarka András , Bánhegyi Gábor
Published: August 2011

Disulfide bond formation in proteins is an effective tool of both structure stabilization and redox regulation. The prokaryotic periplasm and the endoplasmic reticulum of eukaryotes were long considered as the only compartments for enzyme mediated formation of stable disulfide bonds. Recently, the mitochondrial intermembrane space has emerged as the third protein-oxidizing compartment. The classic view on the mechanism of oxidative folding in the endoplasmic reticulum has also been reshaped by new observations. Moreover, besides the structure stabilizing function, reversible disulfide bridge formation in some proteins of the endoplasmic reticulum, seems to play a regulatory role. This review briefly summarizes the present knowledge of the redox systems supporting oxidative folding, emphasizing recent developments.
Depression in adolescents with chronic medical illness
Authors: Rao Chandrika , Ramu Somashekar Ankanahalli , Maiya Parampalli Padmanabha
Published: August 2011

Chronic illness can lead to anxiety and depression in both parents and their adolescents. The identification of predictor variables would greatly assist pediatricians and parents in identifying children at risk for prolonged depression. Findings indicate that age and gender of the adolescent and duration of illness did not affect the depression score, but type of illness affected the score significantly. This article illustrates the fact that pediatricians can use the Beck Depression Inventory to identify depression and guide parents and adolescents to adequate medical care.
Potentiometric and spectrophotometric characterization of the UO2
2+-citrate complexes in aqueous solution, at different concentrations, ionic strengths and supporting electrolytes
Authors: Berto Silvia , Crea Francesco , Daniele Pier G. , De Stefano Concetta , Prenesti Enrico , Sammartano Silvio
Published: August 2011

In this paper we report an investigation on the interactions between dioxouranium(VI) and citrate using potentiometry (H+-glass electrode) and UV-spectrophotometry. Potentiometric measurements were carried out in NaCl and KNO3 aqueous solutions at t = 25 ºC in a wide range of experimental conditions (concentrations, ligand/metal molar ratio, pH, titrants). Measurements in NaCl were carried out at different ionic strength values (0.1 ≤ I/mol L−1 ≤ 1.0); different procedures were employed for the acquisition of experimental data and careful analysis of these data performed. In all cases the speciation model that best fits experimental data takes into account the formation of the following species: UO2(Cit)−, (UO2)2(Cit)2
2−, (UO2)2(Cit)2(OH)
2
4−
, (UO2)2(Cit)2(OH)3−, (UO2)2(Cit)(OH)2
−, (UO2)2(Cit)(OH)0, (UO2)3(Cit)2(OH)
5
5−.

The dependence on ionic strength of formation constants was taken into account by using both a simple Debye–Hückel type equation and the SIT (specific ion interaction theory) approach.

Moreover, a visible absorption spectrum for each complex reaching a significant percentage of formation in solution (KNO3 medium) has been calculated to characterise the compounds found by pH-metric refinement.

Recommended values for the uranyl-citrate species were proposed for each ionic strength values in NaCl aqueous solution. Comparison with literature stability constants is reported too.
Reflexões e contribuições para o Plano Integrado de Ações de Vigilância em Saúde do Ministério da Saúde (MS) de Populações Expostas a Agrotóxicos
Authors: Moisés Marcia , Machado Jorge Mesquita Huet , Peres Frederico , Hennington Élida , Beltrami Aramis Cardoso , Beltrami Neto Adelcki Camilo
Published: August 2011

Este artigo enuncia a problemática da contaminação humana e ambiental devido ao uso abusivo e indevido de agrotóxicos e suas conseqüências para saúde humana e para o ambiente, e tem como objetivo apresentar reflexões e contribuições para o Plano Integrado de Ações de Vigilância em Saúde do Ministério da Saúde de Populações Expostas a Agrotóxicos (Plano Integrado do MS) oriundas do Projeto Avaliação e Controle da Exposição Humana e Ambiental do Distrito Federal (Projeto DF). A metodologia utilizada privilegiou a leitura e a análise dos relatórios e memórias do Plano Integrado do MS e do Projeto DF, como também a observação participante em reuniões preliminares de elaboração do Plano na Coordenação Geral de Vigilância em Saúde Ambiental (CGVAM) e na Oficina "A integração entre a Vigilância em Saúde do Trabalhador e a Vigilância Sanitária: a questão da informação"coordenada pelo GT de Saúde do Trabalhador da Associação Brasileira de Saúde Coletiva (Abrasco) e nas reuniões e oficinas do Projeto DF. Os resultados das discussões sugerem que a estratégia de investigação utilizada na sua construção revela-se como um aporte propositivo e que o Projeto DF pode constituir-se como experiência piloto para o Plano Integrado do MS.
Life history theory predicts fish assemblage response to hydrologic regimes
Authors: Mims Meryl C. , Olden Julian D.
Published: August 2011

The hydrologic regime is regarded as the primary driver of freshwater ecosystems, structuring the physical habitat template, providing connectivity, framing biotic interactions, and ultimately selecting for specific life histories of aquatic organisms. In the present study, we tested ecological theory predicting directional relationships between major dimensions of the flow regime and life history composition of fish assemblages in perennial free‐flowing rivers throughout the continental United States. Using long‐term discharge records and fish trait and survey data for 109 stream locations, we found that 11 out of 18 relationships (61%) tested between the three life history strategies (opportunistic, periodic, and equilibrium) and six hydrologic metrics (two each describing flow variability, predictability, and seasonality) were statistically significant (P ≤ 0.05) according to quantile regression. Our results largely support a priori hypotheses of relationships between specific flow indices and relative prevalence of fish life history strategies, with 82% of all significant relationships observed supporting predictions from life history theory. Specifically, we found that (1) opportunistic strategists were positively related to measures of flow variability and negatively related to predictability and seasonality, (2) periodic strategists were positively related to high flow seasonality and negatively related to variability, and (3) the equilibrium strategists were negatively related to flow variability and positively related to predictability. Our study provides important empirical evidence illustrating the value of using life history theory to understand both the patterns and processes by which fish assemblage structure is shaped by adaptation to natural regimes of variability, predictability, and seasonality of critical flow events over broad biogeographic scales.
Collapse and reorganization of a food web of Mwanza Gulf, Lake Victoria
Authors: Downing Andrea S. , van Nes Egbert H. , Janse Jan H. , Witte Frans , Cornelissen Ilse J. M. , Scheffer Marten , Mooij Wolf M.
Published: August 2011

Lake Victoria in East Africa is the world's second largest freshwater system. Over the past century the ecosystem has undergone drastic changes. Some 30 years after the introduction of Nile perch (Lates niloticus) and Nile tilapia (Oreochromis niloticus) in the 1950s, the highly diverse community of native haplochromines collapsed, leaving a system dominated by only four species: the native cyprinid dagaa (Rastrineobola argentea) and shrimp (Caridina nilotica), as well as the introduced Nile perch and Nile tilapia. More recently, an unexpected resurgence of haplochromines has been reported. To understand these changes in terms of ecosystem functioning and of changes in growth of trophic groups, we created mass balances of the food web near Mwanza, Tanzania, before, during, and after the Nile perch boom (1977, 1987, and 2005), using the application ECOPATH. We connected these mass balances with a dynamic model assuming linear trends in net growth rates of the trophic groups. Our analysis suggests that the Nile perch boom initially altered the biomass distribution over trophic levels. Also, results indicate that not only fishing but also changes at the detritivores' trophic level might have played an important role in driving changes in the system. Both the mass balances and the dynamic model connecting them reveal that, after a major distortion during the Nile perch boom, the biomass distribution over the main trophic levels had largely recovered its original (1977) state by 2005. However, no such return appeared in terms of community structure. Biodiversity in the new state is dramatically lower, consisting of introduced species and a few native surviving species. We conclude that at an aggregate level Lake Victoria's ecosystem has proved to be resilient in the sense that its overall trophic structure has apparently recovered after a major perturbation. By contrast, its intricate functional structure and associated biodiversity have proved to be fragile and seem unlikely to recover.
Weighted Lasso with Data Integration
Authors: Bergersen Linn Cecilie , Glad Ingrid K. , Lyng Heidi
Published: August 2011

The lasso is one of the most commonly used methods for high-dimensional regression, but can be unstable and lacks satisfactory asymptotic properties for variable selection. We propose to use weighted lasso with integrated relevant external information on the covariates to guide the selection towards more stable results. Weighting the penalties with external information gives each regression coefficient a covariate specific amount of penalization and can improve upon standard methods that do not use such information by borrowing knowledge from the external material. The method is applied to two cancer data sets, with gene expressions as covariates. We find interesting gene signatures, which we are able to validate. We discuss various ideas on how the weights should be defined and illustrate how different types of investigations can utilize our method exploiting different sources of external data. Through simulations, we show that our method outperforms the lasso and the adaptive lasso when the external information is from relevant to partly relevant, in terms of both variable selection and prediction.
Modeling of Respiration Rate of Capsicum at Various Temperatures
Authors: Pandey Sushil Kumar , Goswami Tridib Kumar
Published: August 2011

Respiration rate is critical in the design of modified atmosphere/control atmosphere (MA / CA) storage system. Success of these systems is based on correctly mapping of respiration process. Respiration rate of capsicum was studied at 5, 10, 15, 20, and 25 °C using closed system respiration method. Respiration was found to be influenced by temperature and followed Arrhenius law. The generated data were used to develop different types of models, one based on principle of enzyme kinetics, and two based on regression analysis. All developed models were validated at 13°C. The obtained results showed that developed models were valid and represented the respiration process. Model based on principle of enzyme kinetics with Arrhenius type and four-parameter second-order polynomial model showed better agreement with the experimental data than the nonlinear model developed.
Programmed cell death proteins and chronic leukemia
Authors: Brajuskovic G. , Strnad Milica , Cerovic Snezana , Romac Stanka
Published: August 2011

Apoptosis or programmed cell death is a genetically regulated process of
cellular suicide. Apoptosis has been implicated in a wide range of
pathological conditions, and mutations in apoptotic genes play important
roles in the process of malignant transformation. Chronic leukemia represents
a neoplastic disorder caused primarily by defective programmed cell death, as
opposed to increased cell proliferation. This paper presents the main results
of our ten-year research on the apoptosis of leukemia cells. The research
included the morphological aspects of the process, the effect of
antineoplastic agents on the induction of apoptosis in leukemia cells and
expression analysis of the proteins involved in programmed cell death.
Special attention was paid to the expression and interaction of the Bcl-2
family of proteins in leukemia cells. The ultimate aim of the study of
apoptosis of leukemic cells is the discovery of new biological agents that
might be used in the treatment of chronic leukemia.
Chemical compositions and antioxidant activities of five endemic Asperula taxa
Authors: Minareci Ersin , Ergönül Bülent , Kayalar Hüsniye , Kalyoncu Fatih
Published: August 2011

In this study, the proximate chemical composition, total flavonoid and
?-tocopherol amounts and antioxidant activities of five endemic Asperula
taxa (Rubiaceae) (A. brevifolia, A. pseudochlorantha var. antalyensis, A.
pseudochlorantha var. pseudochlorantha, A. purpurea subsp. apiculata and A.
serotina) were determined. Antioxidant properties of methanol extracts were
studied by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging
method. Among the all Asperula extracts A. brevifolia showed the most potent
radical scavenging activities.
The effect of inoculation time and inoculum concentration on the productive response of Tetraselmis chuii (Butcher, 1958) mass cultured in F/2 and 2-F media
Authors: López-Elías José , Esquer-Miranda Edgard , Martínez-Porchas Marcel , Garza-Aguirre María , Rivas-Vega Martha , Huerta-Aldaz Nolberta
Published: August 2011

Diverse protocols have been used in the shrimp industry to produce
microalgae. In this work, we evaluated the effect of inoculation time (0600
and 1200) and initial microalgae density (0.4 and 0.8 x105cells mL-1) in
static outdoor cultures of Tetraselmis chuii, using F/2 and 2-F media. A
significant effect on cell density (> 6.7 x 105 cells mL-1) was found after
treatments with a high initial density and in the F/2 medium. The higher
productive responses were observed in treatments inoculated at 0600. The
better productive response was found in treatments inoculated at 0600 with
0.8 x 105 cells?mL-1 in the F/2 medium.
The protective effect of curcumin against sodium fluoride-induced oxidative stress in rat heart
Authors: Nabavi S.F. , Nabavi S.M. , Ebrahimzadeh M.A. , Eslami Sh. , Jafari N. , Moghaddam Hajizadeh
Published: August 2011

In the present study the cardioprotective effects of curcumin, a herbal
polyphenolic compound, against sodium fluoride (NaF)-induced toxicity in rat
heart was evaluated. Fifty rats were divided into five experimental groups
containing 10 rats each. Group I received standard water and diet and was
used as a normal group; groups II and III were pretreated with curcumin
intraperitoneally for 7 days prior to NaF intoxication. Group IV was
pretreated with vitamin C, a standard antioxidant, intraperitoneally for 7
days prior to NaF intoxication and used as a positive control group. The
animals in group V were intoxicated with NaF for the same time and used as a
control group. There was a significant increase in lipid peroxidation along
with a decrease in superoxide dismutase activity in the homogenates of
tissues of the NaF-treated animals. Curcumin pretreatment in animals prior to
fluoride intoxication normalized the levels of biochemical parameters
measured.
Biogenic amines in protocerebral A2 neurosecretory neurons of Lymantria dispar L. (Lepidoptera:Lymantriidae): Response to trophic stress
Authors: Peric-Mataruga Vesna , Mrdakovic Marija , Vlahovic Milena , Ilijin Larisa , Tomanic Jankovic , Mircic D. , Nenadovic Vera
Published: August 2011

The number, morphometric parameters and amount of aminergic neurosecretory
product of protocerebral A2 neurosecretory neurons were investigated in the
fifth instar of Lymantria dispar caterpillars, following a suitable or
unsuitable trophic regime. Caterpillars originated from two populations
(Quercus rubra or Robinia pseudoacacia forest) and were differently adapted
to trophic stress, i.e. feeding on locust tree leaves - unsuitable host
plant. The number of neurosecretory neurons was higher in the caterpillars
originated from Robinia population than in Quercus population, regardless of
feeding. A2 neurosecretory neurons, nuclei and their nucleoli were larger in
caterpillars fed with unsuitable leaves in both populations. There was more
aminergic product in the A2 neurosecretory neurons of the caterpillars fed
with unsuitable leaves independently of population origin.
Differential in vivo regulation of TH and DBH mRNA in rat atria by maprotiline and fluoxetine
Authors: Spasojevic Natasa , Jovanovic P. , Spasojevic-Tisma Vera , Djelic N. , Dronjak Sladjana
Published: August 2011

It is well known that antidepressants affect central monoaminergic
neurotransmission and that they also modulate hormone release in peripheral
tissues. Repeated maprotiline (a noradrenaline reuptake inhibitor) and
fluoxetine (a serotonin reuptake inhibitor) treatment on gene expression of
the catecholamine biosynthetic enzymes were examined in rat atria and
ventricles in vivo. Maprotiline decreased the gene expression of tyrosine
hydroxylase (TH) and dopamine?-hydroxylase (DBH) in the rat atrium.
Fluoxetine increased gene expression of TH and DBH, but not of
phenylethanolamine N-methyltransferase (PNMT). Chronic application of
antidepressants did not change the expression of these enzymes in the
ventricles. We conclude that repeated administration of fluoxetine enhances
gene transcription of TH and DBH and subsequently stimulates noradrenaline
synthesis in rat atria in vivo.
Chromosome instability in Alzheimer’s disease
Authors: Spremo-Potparevic B. , Zivkovic L. , Plecas-Solarovic B. , Bajic V.P.
Published: August 2011

Alzheimer?s disease (AD), as the most common form of dementia, has for many
years attracted the attention of researchers around the world, primarily
because of the problems of reliable diagnostic methods that could help in the
early detection of this devastating disease. One of the important aspects of
genetic research related to AD is the analysis of chromosome instability
which includes: aneuploidies of different chromosomes, telomere shortening
and the phenomenon of premature centromere division (PCD). The aim of this
study was to describe specific biomarkers in different types of cells as
potential parameters for the diagnosis of AD in order to promptly recognize
pre-symptomatic stages and prevent the development of disease and/or slow
down its progression.