Characterization of the free energy spectrum of peptostreptococcal protein L

BACKGROUND: Native state hydrogen/deuterium exchange studies on cytochrome c and RNase H revealed the presence of excited states with partially formed native structure. We set out to determine whether such excited states are populated for a very small and simple protein, the IgG-binding domain of peptostreptococcal protein L. RESULTS: Hydrogen/deuterium exchange data on protein L in 0-1.2 M guanidine fit well to a simple model in which the only contributions to exchange are denaturant-independent local fluctuations and global unfolding. A substantial discrepancy emerged between unfolding free energy estimates from hydrogen/deuterium exchange and linear extrapolation of earlier guanidine denaturation experiments. A better determined estimate of the free energy of unfolding obtained by global analysis of a series of thermal denaturation experiments in the presence of 0-3 M guanidine was in good agreement with the estimate from hydrogen/deuterium exchange. CONCLUSIONS: For protein L under native conditions, there do not appear to be partially folded states with free energies intermediate between that of the folded and unfolded states. The linear extrapolation method significantly underestimates the free energy of folding of protein L due to deviations from linearity in the dependence of the free energy on the denaturant concentration.     

Haematopoietic response and bcl-2 expression in patients with acute myeloid leukaemia

In the Bethesda System for reporting cervicovaginal cytology results, 1 criterion for smear adequacy is an adequate squamous component. The accuracy of a cytologist's estimate that 10% of the slide is covered by squamous cells, the adequacy threshold, has not been determined. The percentage of the surface of a glass slide covered by squamous cells was independently estimated by 4 cytologists on 2 occasions by microscopic examination of 83 buccal smears prepared to display estimated coverage of 1% to 20% of the slide surface. The accuracy of visual estimates was compared with measurements by the TracCell System. Each observer made a third set of estimates after receiving 5 slides with known coverage. Median coverage by visual estimation ranged from 4% to 25%, but as measured by the TracCell system was 2%. Median estimated coverage was significantly different for 2 of 4 observers between first and second viewings and between all but 1 pair of observers. For all observers, it was significantly higher than the true coverage. A visual estimate of 10% coverage corresponded to a true median coverage of 3%. When provided with a physical standard, the median estimated coverage by 3 of 4 observers was not statistically different from the true coverage, and interobserver kappa values improved. Unaided visual estimation of the adequacy of squamous cell coverage is neither reproducible nor accurate. What most cytologists consider "adequate" coverage represents only 3% coverage. The availability of a physical standard dramatically increases reproducibility and accuracy.     

Function changes in spinal muscular atrophy II and III. The DCN/SMA Group

The course of spinal muscular atrophy (SMA) is not well established except for those patients whose age of onset is before 6 months and who achieve only "sit with support" as their maximum function (Werdnig-Hoffmann disease or SMA I). This study shows that there is another group of SMA patients whose age of onset and maximum function achieved can be used as prognostic guides. Fifty percent of SMA patients who could walk without assistance and whose onset was prior to age 2 years lost the ability to walk independently by age 12. Fifty percent of SMA patients who walked and whose onset was between 2 and 6 years of age lost walking ability by age 44 years. Fifty percent of SMA patients who could walk with assistance as their best function ever achieved lost this ability by age 7 years, unrelated to age of onset; none could walk with assistance after age 14 years. Seventy-five percent of SMA patients who developed the ability to sit independently as their best function were still sitting after age 7 years independent of age of onset; 50% of this group could sit independently after age 14 years. Eighty-five percent of SMA patients who could walk could not negotiate stairs without holding onto a rail. They could raise their hands above the head; however, as they lost walking ability, they lost this function as well. Only one SMA patient whose maximum function was sitting independently could get to the sitting position on his own. Only two of these patients could hold their hands above their heads. All patients with SMA lose function over time. This function loss occurs slowly and is related primarily to maximum function achieved; knowledge of age of onset provides helpful information, especially for predicting the loss of independent walking.     

Characterization of HEp-2 cell projection formation induced by diffusely adherent Escherichia coli

Diffusely adherent Escherichia coli (DAEC) are diarrheagenic E. coli whose pathogenetic mechanisms are largely unknown. DAEC have been shown to induce an unusual phenotype upon adherence to HEp-2 cells in culture characterized by the induction of long thin membrane processes extending from the cell surface. In addition, DAEC have been shown to be protected from the bactericidal effects of gentamicin when incubated with HEp-2 cells. In our studies, we found that three DAEC strains induced formation of eukaryotic cell processes and were protected from gentamicin killing after a 3 h incubation. Preincubation of HEp-2 cells with colchicine or cytochalasin D prior to infection with DAEC strain C1845 resulted in decreased projection formation, suggesting that the effect was dependent upon microfilament and microtubule rearrangement. When the standard gentamicin protection assay was extended for an additional 3 h incubation in the presence of gentamicin, a greater number of DAEC survived gentamicin treatment, more eukaryotic projections were seen in association with the bacteria and the bacteria were actually observed to be "embedded' within these projections. Projection formation was not observed when the bacteria were separated from the cells by a permeable membrane or when the inoculum was inactivated by ultraviolet irradiation. Transposon TnphoA mutants of C1845 were screened for decreased gentamicin protection. All three mutants which were deficient in gentamicin protection demonstrated less projection formation. Insertion mutations affecting gentamicin protection were localized to both the chromosome (two) and a plasmid (one). Eukaryotic projections are a novel interaction of DAEC with epithelial cells, may play a role of the survival of the bacteria against host defenses and may contribute to DAEC pathogenesis. The effect is dependent upon epithelial cell contact and requires multiple bacterial genes.     

Fine needle aspiration biopsy of the salivary glands. A five-year experience with emphasis on diagnostic pitfalls

OBJECTIVE: To review the fine needle aspiration (FNA) findings in 151 patients who presented with salivary gland (both major and minor) enlargement from January 1991 to December 1995 in order to determine its sensitivity and specificity and to study the various pitfalls. STUDY DESIGN: The study group consisted of 77 males and 74 females, 16-98 years old (average 55). One hundred twenty-five aspirates (83%) were from the parotid gland, 23 (15%) from the submandibular gland and 3 (2%) from the soft palate. One hundred thirty-seven cases (91%) were adequate for diagnosis. There were 89 (59%) aspirations done by cytopathologists, 100% of which were diagnostic, and 62 (41%) done by clinicians, 48 (77%) of which were diagnostic. Sixty-eight (45%) cases had histologic confirmation. There were 104 (75.9%) benign, 20 (14.6%) malignant and 13 (9.5%) atypical cytologic diagnoses. RESULTS: Using histology as the "gold standard," the sensitivity of FNA cytology was 91%, with a specificity of 96%. A number of problems were encountered in interpreting some cases, not only in differentiating benign from malignant ones but also in the specific classification of these neoplasms. Problems encountered involved differentiating hematopoietic from non-hematopoietic lesions and interpretation of spindle cell neoplasms, acinic cell carcinoma, mucoepidermoid carcinoma, adenoid cystic carcinoma, lymphoproliferative disorders, postirradiation changes, sialadenitis and atypia in pleomorphic adenoma. CONCLUSION: FNA biopsy of the salivary gland is a sensitive and specific diagnostic tool at our institution. Particular attention to subtle morphologic changes may aid in avoiding pitfalls and arriving at the right diagnosis.     

Spectral, magnetic and biological studie on some bivalent 3d metal complexes of hydrazine derived schiff-base ligands

Metal(II) complexes of hydrazine derived Schiff-base ligands of the type M(L)(2)Cl(2) where M = Co, Cu, Ni and Zn and L = L(1) and L(2) have been prepared and characterised by molar conductance, magnetic moment, elemental analysis and electronic, IR, H-NMR and (13)C spectral data.The different modes of chelation of the ligands and their comparative biological properties against different bacterial species are reported.     

Thermal analysis of peritectic alloys

Predictions that increasing cooling rate will result in increasing elevation of the apparent peritectic arrest temperature have been confirmed for Cd-Ag and Zn-Cu alloys in line with observations of Al-Ti alloys. The shape of the peritectic cooling curve is related to the competition between the peritectic transformation and direct crystallization from the melt in the formation of the secondary phase. The effect on solidification of the relative depressions of the and liquidus during cooling is considered.