Distinct autophosphorylation sites sequentially produce autonomy and inhibition of the multifunctional Ca2+/calmodulin-dependent protein kinase

The multifunctional Ca2+/calmodulin-dependent protein kinase (multifunctional CaM kinase) may be an important mediator for neurotransmitters and hormones that utilize Ca2+ as a "second messenger." We examined the ability of autophosphorylation to convert the multifunctional CaM kinase to a Ca2+/calmodulin-independent (autonomous) form to determine whether autophosphorylation is a mechanism for short- or long-term enhancement of Ca2+ action. As the kinase incorporates phosphate during continuous stimulation by Ca2+/calmodulin, its ability to phosphorylate exogenous substrates becomes increasingly autonomous. Withdrawal of Ca2+ after a critical level of phosphate incorporation is reached leads to a "burst" or rapid increase in Ca2+-independent autophosphorylation. The "burst" of autophosphorylation is distinct from the initial Ca2+-dependent autophosphorylation, however, since it inhibits substrate phosphorylation. Both Ca2+-dependent and Ca2+-independent substrate phosphorylation are inhibited by this autonomous autophosphorylation. Thus, autophosphorylation has a dual role in modulating the activity of multifunctional CaM kinase. It initially enables the kinase to continue phosphorylating substrates after Ca2+ levels decline, but it eventually suppresses this autonomous activity. Tryptic phosphopeptide mapping demonstrates that appearance of phosphothreonine-containing peptides is common to several conditions used to generate an autonomous enzyme. Sequencing reveals the critical "autonomy" site to be threonine286. The inhibitory mode of autophosphorylation involves 3 additional phosphopeptides containing a serine and a threonine residue.     

Injuries to the profunda femoris artery and its branches

Six cases of uncommon profunda femoris artery injury are analyzed. One half of the cases were not treated surgically at the time of injury and resulted in chronic arteriovenous fistulas. Even after operative correction, residual symptoms persisted in two of these three cases. The generous use of arteriography is recommended in cases of proximal thigh penetrating injuries to avoid this problem. Either repair or ligation of the injured vessel yielded the same good results. The postoperative hospital stay was short in all instances, ranging from four to eight days, and there were no deaths.     

Antigen presentation is a function of all B cell subpopulations separated on the basis of size

Purified splenic B cells from nonimmune mice were separated by counterflow centrifugal elutriation into 6 subpopulations containing cells of discrete sizes ranging from 119 to 200 μm³. B cells of each subpopulation were competent to process and present a native globular protein antigen, cytochrome c, to a cytochrome c-specific T cell hybrid. In all cases, the B cells' antigen-presenting function was radiation sensitive and did not require T cells or T cell products, since B cells fixed with paraformaldehyde effectively presented a carboxyl-terminal peptide fragment of cytochrome c containing the T cell determinant. Furthermore, the antigen-presenting function of B cells of each subpopulation was augmented by treatment with submitogenic doses of the F(ab')₂ fragment of rabbit anti-mouse Ig antibodies, in that 10-30-fold fewer B cells were required and higher maximal T cell responses were achieved, indicating that B cells of all sizes are capable of being regulated in their antigen presentation function through their surface Ig. In addition, B cells of each subpopulation responded to soluble factors present in the supernatants of activated T cells as evidenced by an increase in volume and by the uptake of [³H]thymidine. These results indicate that B cells, regardless of size, are able to participate in at least two essential phases of T cell-dependent antibody responses, initiating the interaction by processing and presenting antigen to helper T cells and responding to soluble helper factors secreted by activated T cells.     

Cr Migration Potential and Species Properties in the Soil Profile from a Chromate Production Site in the Groundwater Depression Cone Area

The relationship between the migration process and speciation distribution of Cr is important for the risk assessment in the underground environment. In this work, soil columns were collected from the chromate production site, with a 40-year operation, in the groundwater depression cone area of North China plain. The relationship between chromium pollution features and the geochemical properties of soil was established, and the migration risk of Cr(VI) was assessed based on the Nemerow composite index and Hydrus-1D model. The maximum total Cr concentration in the chromium slag dumping site reached 907 mg/kg, and that in the chromate production workshop was more than 200 mg/kg across the depth. The migration of Cr might be accelerated in the soil with abundant Mn (236–1461 mg/kg) but scarce organic matters (<?0.45%). The Hydrus simulation indicated that Cr(VI) would reach a cumulative flux of 300–729 mg/cm² after 50 years.

     

Characterisation of the IgE-binding immunodominant epitopes on Ara h1

Peanut allergy is one of the most severe food allergies due to its persistency and life-threatening character. Serum IgE from patients with documented peanut hypersensitivity reactions and synthetic peptides were used to screen the linear IgE-binding epitopes on the major peanut allergen, Ara h 1. Five major epitopes that bound peanut-specific serum IgE from more than 60% of patients tested were identified. Mutational analysis of the immunodominant epitope showed that single amino acid changes had dramatic effects on IgE-binding characteristics. Mapping and characterisation of the IgE-binding epitopes on Ara h1 could be used in future immunotherapeutic approaches for peanut allergy disease.     

The expression of placental alkaline phosphatase (PLAP) and PLAP-like enzymes in normal and neoplastic human tissues. An immunohistological survey using monoclonal antibodies

The immunohistological expression of placental alkaline phosphatase (PLAP) and PLAP-like enzyme was studied in frozen sections from a wide variety (n = 254) of normal and malignant tissues using monoclonal antibodies reactive with PLAP (H317) and PLAP/PLAP-like enzyme (H17E2; H315). PLAP/PLAP-like reactivity was seen in normal thymus, and foetal and neonatal testis, and in 21 out of 22 malignant germ cell tumours (GCTs), but was also found in normal endocervix, normal Fallopian tube and in 28 out of 167 non-GCTs (particularly in ovarian and proximal gastrointestinal tract tumours). Positivity for true PLAP (as demonstrated with H317) was seen in term placenta, in endocervix, and in Fallopian tube (but not in other normal tissues) and was commonly found in ovarian and proximal gastrointestinal tract tumours. Reactivity with H317 was unusual in malignant GCTs (2 out of 22 cases). These findings confirm that PLAP/PLAP-like positivity is a highly sensitive immunohistological marker for malignant GCTs, but one which by itself is of only moderate specificity. Furthermore, expression of true PLAP is rare in GCTs and favours instead an origin from the ovary or proximal gastrointestinal tract. The results also indicate that the predominant heat-stable alkaline phosphatase species in normal foetal and neonatal testis, and in thymus has a similar immunohistological profile to that found in malignant GCTs, and is a PLAP-like enzyme ("germ cell alkaline phosphatase") distinct from true PLAP. The occurrence of this marker in GCTs would appear to reflect increased eutopic production of an enzyme present in trace amount in corresponding normal tissues rather than a genuine example of ectopic expression.     

三磷酸腺苷生物荧光液闪测定法检测活细胞计数

为建立一种检测活细胞数的方法,作者采用三磷酸腺苷(ATP)生物荧光液闪测定法检测小鼠纤维瘤细胞株L929.ATP标准曲线工作范围为10 -9～10-5mol/ml,相关系数r=0.9963(P＜0.001);当活细胞数在3×1 02～106个/ml时与发光计数值有良好的线性关系,r=0.9922(P＜0.001),变异系数 CV = 1%～3%.用ATP生物荧光液闪测定法检测活细胞数灵敏、简便、准确、重复性好,用液闪计数仪即可完成测定,有较大的实用价值.