Serratiopeptidase Affects the Physiology of Pseudomonas aeruginosa Isolates from Cystic Fibrosis Patients

Pseudomonas aeruginosa is frequently involved in cystic fibrosis (CF) airway infections. Biofilm, motility, production of toxins and the invasion of host cells are different factors that increase P. aeruginosa’s virulence. The sessile phenotype offers protection to bacterial cells and resistance to antimicrobials and host immune attacks. Motility also contributes to bacterial colonization of surfaces and, consequently, to biofilm formation. Furthermore, the ability to adhere is the prelude for the internalization into lung cells, a common immune evasion mechanism used by most intracellular bacteria, such as P. aeruginosa. In previous studies we evaluated the activity of metalloprotease serratiopeptidase (SPEP) in impairing virulence-related properties in Gram-positive bacteria. This work aimed to investigate SPEP’s effects on different physiological aspects related to the virulence of P. aeruginosa isolated from CF patients, such as biofilm production, pyoverdine and pyocyanin production and invasion in alveolar epithelial cells. Obtained results showed that SPEP was able to impair the attachment to inert surfaces as well as adhesion/invasion of eukaryotic cells. Conversely, SPEP’s effect on pyocyanin and pyoverdine production was strongly strain-dependent, with an increase and/or a decrease of their production. Moreover, SPEP seemed to increase swarming motility and staphylolytic protease production. Our results suggest that a large number of clinical strains should be studied in-depth before drawing definitive conclusions. Why different strains sometimes react in opposing ways to a specific treatment is of great interest and will be the object of future studies. Therefore, SPEP affects P. aeruginosa’s physiology by differently acting on several bacterial factors related to its virulence.     

The roles of the N-linked glycans and extension regions of soybean beta-conglycinin in folding, assembly and structural features

Beta-conglycinin, one of the dominant storage proteins of soybean, is a trimer composed of three subunits, alpha, alpha' and beta. All subunits are N-glycosylated and alpha and alpha' contain extension regions in addition to the core regions common to all subunits. Non-glycosylated individual subunits and deletion mutants (alpha(c) and alpha'(c)) lacking the extension regions of alpha and alpha' were expressed in Escherichia coli. All recombinant proteins were purified to near homogeneity and appeared to have the correct conformation, as judged by CD, density-gradient centrifugation and gel-filtration profiles, indicating that the N-linked glycans and extension regions are not essential for the folding and the assembly into trimers of beta-conglycinin. Density-gradient centrifugation, gel-filtration and differential scanning calorimetry profiles of the recombinant proteins and the native beta-conglycinin indicated that the N-linked glycans and extension regions contribute to the dimension of beta-conglycinin but not to the density and the thermal stability. Comparing the solubilities of the individual subunits with those of deletion mutants, only the alpha and alpha' subunits were soluble at lower ionic strength (mu < 0.25) at around the pH value of the endoplasmic reticulum. This suggests that the extension regions play an important role in the prevention of aggregation in the endoplasmic reticulum in analogy with the N-linked glycans.     

Prostate-specific antigen concentrations in serum in acute illnesses

In light of recent studies showing that prostate-specific antigen (PSA) complexes with certain blood proteins, we studied the effects of acute-phase reactants and alpha 2-macroglobulin (A2MG) on serum concentrations of PSA. Serum samples were obtained from 419 men admitted to an acute-care facility. Various acute-phase reactants-including C-reactive protein, alpha 1-acid glycoprotein, alpha 1-antitrypsin, and alpha 1-antichymotrypsin-and A2MG were measured with a Beckman Array analyzer in parallel with determinations of PSA concentrations by two methods, the Hybritech Tandem RIA and the Abbott PSA IMx. Evaluation by Spearman rank correlation revealed a significant negative correlation of A2MG with PSA values (P < 0.01) and (as expected) a positive correlation of age with PSA values (P < 0.001). The former correlation suggests the possibility that patients with high serum concentrations of A2MG may give falsely decreased results for PSA concentrations in serum.     

Transposition of rotameter tubes

We report a case of a flexor sheath tenosynovitis caused by Mycobacterium marinum.     

Historical evolution of the treatment of esophageal cancer at the Hospital A. C. Camargo, S?o Paulo, Brazil

OBJECTIVES: From 1947-1986 we reviewed a historical series of 1,900 cases of esophageal cancers registered at the A.C. Camargo Hospital, S?o Paulo, Brazil. Two hundred and thirty four cases were submitted to surgical resection. During these 4 decades the treatment philosophy of these tumors has changed. METHOD: Five different historical groups were identified and the results are presented. RESULTS: The first group (1947-60) consisted of 47 cases only submitted to surgical resection. The second group (1961-70) of 56 cases had pre and pos surgery radiotherapy in low doses and the reconstruction was made using subcutaneous colon. From 1971-75 the same approach was used except with high dose preoperative radiotherapy (31 cases). In the 4th group (1976-82) of 68 cases preoperative radiotherapy (high dose) and chemotherapy were used. In the last group (1983-86) composed of 32 cases the treatment was preoperative chemotherapy, surgical resection with gastric reconstruction followed with high doses radiotherapy in the surgical bed and chemotherapy. The only significant prognostic factors in the statistical analysis were tumor size and involvement peri-esophageal lymph nodes. CONCLUSION: A five year survival from 3.7% to 9.0% was obtained through the use of the fifth group treatment planning.