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1.  Amyloidosis: an unusual cause of portal hypertension 
Autopsy & Case Reports  2016;6(2):9-18.
Amyloidosis comprises a group of diseases that occurs in five to nine cases per million patients per year worldwide irrespective of its classification. Although the hepatic involvement in primary amyloidosis is frequent, the clinical manifestations of liver amyloidosis are mild or even absent. The authors report the case of an aged man who complained of diffuse abdominal pain and marked weight loss and presented clinical signs of hepatopathy. Clinical workup revealed portal hypertension with ascites, hemorrhoids, and esophageal varices. The laboratory tests showed the cholestatic pattern of liver enzymes, hyperbilirubinemia, renal insufficiency and massive proteinuria accompanied by the presence of serum pike of monoclonal lambda light chain protein. The outcome was unfavorable, and the patient died. The autopsy findings revealed the diagnosis of amyloidosis predominantly involving the liver and kidneys. The bone marrow examination demonstrated the deposition of amyloid material associated with clonal plasma cells infiltration. The authors call attention to portal hypertension as a rare manifestation of primary amyloidosis. Meanwhile, this diagnosis should be taken into account whenever the hepatopathy is accompanied by laboratory abnormalities consistent with hepatic space-occupying lesions concomitantly with other organs involvement. In the case reported herein, kidney involvement was also present with renal failure, massive proteinuria with monoclonal serum gammopathy, what reinforced the diagnostic possibility of primary amyloidosis.
doi:10.4322/acr.2016.035
PMCID: PMC4982779  PMID: 27547738
Amyloidosis; Liver Diseases; Hypertension, Portal; Multiple myeloma
2.  The recombinant pea defensin Drr230a is active against impacting soybean and cotton pathogenic fungi from the genera Fusarium, Colletotrichum and Phakopsora 
3 Biotech  2016;6(1):59.
Plant defensins are antifungal peptides produced by the innate immune system plants developed to circumvent fungal infection. The defensin Drr230a, originally isolated from pea, has been previously shown to be active against various entomopathogenic and phytopathogenic fungi. In the present study, the activity of a yeast-expressed recombinant Drr230a protein (rDrr230a) was tested against impacting soybean and cotton fungi. First, the gene was subcloned into the yeast expression vector pPICZαA and expressed in Pichia pastoris. Resulting rDrr230a exhibited in vitro activity against fungal growth and spore germination of Fusarium tucumaniae, which causes soybean sudden death syndrome, and against Colletotrichum gossypii var. cephalosporioides, which causes cotton ramulosis. The rDrr230a IC50 corresponding to inhibition of fungal growth of F. tucumaniae and C. gossypii var. cephalosporioides was 7.67 and 0.84 µM, respectively, demonstrating moderate activity against F. tucumaniae and high potency against C. gossypii var. cephalosporioides. Additionally, rDrr230a at 25 ng/µl (3.83 µM) resulted in 100 % inhibition of spore germination of both fungi, demonstrating that rDrr230a affects fungal development since spore germination. Moreover, rDrr230a at 3 µg/µl (460.12 µM) inhibited 100 % of in vitro spore germination of the obligatory biotrophic fungus Phakopsora pachyrhizi, which causes Asian soybean rust. Interestingly, rDrr230a substantially decreased the severity of Asian rust, as demonstrated by in planta assay. To our knowledge, this is the first report of a plant defensin active against an obligatory biotrophic phytopathogenic fungus. Results revealed the potential of rDrr230a as a candidate to be used in plant genetic engineering to control relevant cotton and soybean fungal diseases.
doi:10.1007/s13205-015-0320-7
PMCID: PMC4752952
Defensin; Pisum sativum; Pichia pastoris; Fusarium tucumaniae; Colletotrichum gossypii var. cephalosporioides; Phakopsora pachyrhizi
3.  Lytic enzyme production optimization using low-cost substrates and its application in the clarification of xanthan gum culture broth 
Food Science & Nutrition  2014;2(4):299-307.
Lytic enzymes are widely used in industrial biotechnology as they are able to hydrolyze the bacterial cell wall. One application of these enzymes is the clarification of the culture broth for the production of xanthan gum, because of its viability in viscous media and high specificity. The screening process for filamentous fungi producing lytic enzymes, the optimization of production of these enzymes by the selected microorganism, and the optimization of the application of the enzymes produced in the clarification of culture broth are presented in this article. Eleven fungal isolates were tested for their ability to produce enzymes able to increase the transmittance of the culture broth containing cells of Xanthomonas campestris. To optimize the secretion of lytic enzymes by the selected microorganism the following variables were tested: solid substrate, initial pH, incubation temperature, and addition of inducer (gelatin). Thereafter, secretion of the enzymes over time of incubation was assessed. To optimize the clarification process a central composite rotational design was applied in which the pH of the reaction medium, the dilution of the broth, and the reaction temperature were evaluated. The isolate identified as Aspergillus tamarii was selected for increasing the transmittance of the broth from 2.1% to 54.8%. The best conditions for cultivation of this microorganism were: use of coconut husk as solid substrate, with 90% moisture, at 30°C for 20 days. The lytic enzymes produced thereby were able to increase the transmittance of the culture broth from 2.1% to 70.6% at 65°C, without dilution and without pH adjustment.
doi:10.1002/fsn3.87
PMCID: PMC4221828  PMID: 25473487
Aspergillus tamarii; gram-negative bacteria; protease; response surface methodology
4.  Sport Injuries in Elite Paralympic Swimmers With Visual Impairment 
Journal of Athletic Training  2013;48(4):493-498.
Context:
Visually impaired athletes sustain overuse injuries in the lower limbs, but the pattern of injuries may vary according to the sport. The characteristics of sports injuries in visually impaired swimmers are unknown.
Objective:
To determine the characteristics and epidemiologic measures (prevalence, clinical incidence, and incidence rate) of sports injuries in visually impaired elite national swimmers and to assess differences among visual classes and between sexes.
Design:
Descriptive epidemiology study.
Setting:
International Paralympic competitions held between 2004 and 2008.
Patients or Other Participants:
Twenty-eight elite, visually impaired swimmers (19 males, 9 females) from the Brazilian Paralympic Team participated in this study. Twelve were visual class S11 (blind swimmers), 12 were S12 (low-vision swimmers), and 4 were S13 (low-vision swimmers).
Intervention(s):
A standardized report form was used to collect data during 5 competitions. This was endorsed by 2 Brazilian sports governing bodies.
Main Outcome Measures:
The characteristics, prevalence, clinical incidence, and incidence rate of injuries were calculated.
Results:
Eighteen athletes reported 41 sports injuries, with a prevalence of 64%, clinical incidence of 1.5 injuries per athlete, and an incidence rate of 0.3 injuries per athlete per competition. Overuse injuries (80%) were more frequent than traumatic injuries (20%). The clinical incidence and prevalence varied according to sex and visual class, but no statistical differences were observed (P > .05). The highest proportion of injuries was in the trunk (46.34%), followed by the upper limbs (34.15%). The shoulders (29.27%) were most affected, followed by the thoracic (21.95%) and lumbar spine (17.07%). Spasm (36.59%) was the most frequent diagnosis, followed by tendinopathy (26.83%).
Conclusions:
Visually impaired swimmers had a relatively high proportion of overuse injuries, predominantly associated with muscle spasm in the spine and tendinopathy in the shoulders. No differences were apparent in injury prevalence and clinical incidence among visual classes or between sexes.
doi:10.4085/1062-6050-48.4.07
PMCID: PMC3718352  PMID: 23768122
epidemiology; disability sport; musculoskeletal injuries
5.  A new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) affects Soybean Asian rust (Phakopsora pachyrhizi) spore germination 
BMC Biotechnology  2011;11:14.
Background
Asian rust (Phakopsora pachyrhizi) is a common disease in Brazilian soybean fields and it is difficult to control. To identify a biochemical candidate with potential to combat this disease, a new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) (CaclXIP) leaves was cloned into the pGAPZα-B vector for expression in Pichia pastoris.
Results
A cDNA encoding a chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) (CaclXIP), was isolated from leaves. The amino acid sequence predicts a (β/α)8 topology common to Class III Chitinases (glycoside hydrolase family 18 proteins; GH18), and shares similarity with other GH18 members, although it lacks the glutamic acid residue essential for catalysis, which is replaced by glutamine. CaclXIP was expressed as a recombinant protein in Pichia pastoris. Enzymatic assay showed that purified recombinant CaclXIP had only residual chitinolytic activity. However, it inhibited xylanases from Acrophialophora nainiana by approx. 60% when present at 12:1 (w/w) enzyme:inhibitor ratio. Additionally, CaclXIP at 1.5 μg/μL inhibited the germination of spores of Phakopsora pachyrhizi by 45%.
Conclusions
Our data suggests that CaclXIP belongs to a class of naturally inactive chitinases that have evolved to act in plant cell defence as xylanase inhibitors. Its role on inhibiting germination of fungal spores makes it an eligible candidate gene for the control of Asian rust.
doi:10.1186/1472-6750-11-14
PMCID: PMC3045311  PMID: 21299880
6.  Expression in Escherichia coli, purification, refolding and antifungal activity of an osmotin from Solanum nigrum 
Background
Heterologous protein expression in microorganisms may contribute to identify and demonstrate antifungal activity of novel proteins. The Solanum nigrum osmotin-like protein (SnOLP) gene encodes a member of pathogenesis-related (PR) proteins, from the PR-5 sub-group, the last comprising several proteins with different functions, including antifungal activity. Based on deduced amino acid sequence of SnOLP, computer modeling produced a tertiary structure which is indicative of antifungal activity.
Results
To validate the potential antifungal activity of SnOLP, a hexahistidine-tagged mature SnOLP form was overexpressed in Escherichia coli M15 strain carried out by a pQE30 vector construction. The urea solubilized His6-tagged mature SnOLP protein was affinity-purified by immobilized-metal (Ni2+) affinity column chromatography. As SnOLP requires the correct formation of eight disulfide bonds, not correctly formed in bacterial cells, we adapted an in vitro method to refold the E. coli expressed SnOLP by using reduced:oxidized gluthatione redox buffer. This method generated biologically active conformations of the recombinant mature SnOLP, which exerted antifungal action towards plant pathogenic fungi (Fusarium solani f. sp.glycines, Colletotrichum spp., Macrophomina phaseolina) and oomycete (Phytophthora nicotiana var. parasitica) under in vitro conditions.
Conclusion
Since SnOLP displays activity against economically important plant pathogenic fungi and oomycete, it represents a novel PR-5 protein with promising utility for biotechnological applications.
doi:10.1186/1475-2859-7-7
PMCID: PMC2362109  PMID: 18334031

Results 1-6 (6)