Puroindolines, for years largely investigated for their involvement in wheat kernel hardness, have recently attracted attention thanks to their possible role as antimicrobial proteins. With the aim to enhance our knowledge of these proteins we studied their localization in the kernel, and their antimicrobial activity in vitro against six different bacterial strains. Immunolocalization showed that both the PINs are strongly concentrated in the aleurone layer, but also highly present in the endosperm. Interestingly we observed that puroindolines not only have the same spatial distribution in the kernel, they are also always found co-localized. Their co-localization suggests that they could cooperate in defending the plant against pathogens. We therefore tested antimicrobial activity of PINA and PINB, and a putative synergism between these proteins. The results showed that the two polypeptides can in vitro inhibit growth of all the bacteria tested; furthermore when combined together they are able to enhance each other's toxicity. In view of their antimicrobial activity and of their natural presence in Triticum aestivum wheat flour, puroindolines look promising antibacterial agents and thus deserve further studies aimed at establishing their possible future applications in fields of food and health care. Since PINs were still detectable in bakery products, these proteins may be promising tools in investigating natural ways of food preservation.
Antimicrobial peptides and proteins are being studied with increasing interest because of their broad range antimicrobial activity. Among plant antimicrobial proteins, the wheat seed polypeptides, puroindoline a and puroindoline b, are particularly interesting because of their established antibacterial activity. In this paper we describe different strategies used to clone His tagged and GST tagged puroindolines obtaining 1.5 mg recombinant protein from 1 l culture. The antimicrobial activity of recombinant and native puroindolines was comparable.
Patients (305 patients with pulmonary tuberculosis) and controls (290 household genetically unrelated contacts) were tested by polymerase chain reaction (PCR) for polymorphisms in the intron 15 and the 5 0 untranslated region of the gene coding for the a3 isoform of the human ATPase gene. Diagnosis of pulmonary tuberculosis was based on chest radiography and sputum smear examination and confirmed by PCR and bacteriological tests. Alleles (two at each site) segregated in the form of four haplotype pairs: 13, 14 (very rare), 23, and 24. The 13/24 (double heterozygous) patients were protected against tuberculosis (OR: 0.15; P: 10 À8 ; CI: 0.08-0.3). The 13/13 vs 13/24 and 23/23 vs 23/24 (double homozygous) patients were susceptible to the disease (OR. 5.8; P: 6 Â 10 À7 ; CI: 2.8-11.9; OR: 4.5; P: 5 Â 10 À7 ; CI: 2.5-8.4, respectively).
Lactoferrin (LF) is a member of the transferrin family of iron-binding glycoproteins. It is also a multifunctional protein of 80 kDa that is synthesized by glandular epithelial cells and secreted into mucosal fluid. High levels of LF are present in colostrom and milk and low levels in tears, saliva, and gastrointestinal and reproductive secretions. Data regarding the antifungal effects of LF are limited. Studies have been performed on Candida albicans, which demonstrated that LF inhibits the growth of this fungus. This study reports the results of experiments carried out in order to evaluate the effects of LF on the growth of 11 fungi, which were isolated from plants and soils. These experiments employed the methods of amended agar utilizing nine different concentration levels of LF (0, 0.001, 0.01, 0.1, 1, 10, 100, 1000, 5000 mg L(-1)). The effects of LF on the growth of these fungi were based on measures of the radial growth of the fungal colonies expressed both as percentage of inhibition and as IC(50) values (the concentration at which the fungal growth was inhibited by 50% relative to controls). LF had no effects on Alternaria alternata, Gliocladium roseum, Fusarium solani and Colletotrichum lindemuthianum. It did, however, inhibit the growth of Aspergillus niger, Trichoderma viride, Sclerotinia sclerotiorum, Sclerotium rolfsii, Rhizoctonia solani and Phoma exigua to the point that their IC(50) values ranged from 31.1 mg L(-1) for S. sclerotiorum to 952 mg L(-1) for T. viride.
Background: Uncooperative children require sedative approach for dental treatment. The aim was to assess the effectiveness of Propofol in “Non-Operating Room Anesthesia” (NORA) for paediatric dental treatment; intraoperative side effects; postoperative side effects; post-discharge effects. Methods: a prospective study, involving 109 uncooperative children undergoing sedation in NORA using Propofol for dental treatment, was performed. Working sessions, success/failure, intraoperative and postoperative side effects, number of treatment; type of procedure were assessed. Parents completed a post-discharge questionnaire on: pain; crying; fever; vomiting; headache; drowsiness; excitability; irritability; ability to eat; drugs and medical care needing. Results: Success: 96.7%. Intraoperative side effects: 33.3%. Postoperative side effects: 6.4%. Statistically significant association between: intraoperative side effects and age (p = 0.001), health status (p = 0.0007), weight (p = 0.038), respectively; intraoperative side effects and number/ type of dental treatment (p = 0.0055) and scaling (p = 0.0001), respectively. For post-discharge questionnaires, statistically significant association between: age and crying (p = 0.0001) and headache (p = 0.002), respectively; health status and crying (p = 0.015) and drugs needing (p = 0.04), respectively; weight and crying (p = 0.0004); extraction and pain (p = 0.0001) and crying (p= 0.0073), respectively; scaling and crying (p = 0.04), excitability and irritability (p = 0.03), respectively. Conclusion: Propofol in NORA was effective with minimal side effects.
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