Intracellular signaling events during positive and negative selection of CD4+CD8+ thymocytes in vitro

We have used in vitro models of thymocyte positive and negative selection in conjunction with selective inhibitors of the TCR-mediated signaling cascade to investigate the intracellular signaling events that mediate these processes. We report that Ro 31.8425, a potent and selective inhibitor of protein kinase C, which blocks the activation of mature T cells in a dose-dependent fashion, has no effect on either positive or negative selection of CD4+8+ thymocytes. In contrast, cyclosporin A fails to prevent negative selection, but inhibits positive selection through a direct effect on developing thymocytes, rather than through the perturbation of stromal cell support. Thus, our data suggest that positive and negative selection may operate via distinct intracellular signaling pathways.     

Fine needle aspiration cytology of oral and pharyngeal lesions. A study of 45 cases

Forty-five patients with oral or pharyngeal swellings were subjected to fine needle aspiration cytology (FNAC) of the mucosal surface over eight years. The age of the patients ranged from 2 to 85 years. The male:female ratio was 25:20. The common sites of involvement were palate (16 cases), cheek (9), pharynx (7) and tonsillar/peritonsillar area (6). Tongue, maxilla, alveolus and lips were less frequently involved. FNAC played an important role in differentiating inflammatory from neoplastic lesions and also benign from malignant neoplasms. Fifteen cases were cytologically diagnosed as benign neoplasms and included pleomorphic adenoma (11 cases), schwannoma (2), odontogenic tumor (1) and benign neoplasm not otherwise specified (1). Sixteen cases were diagnosed as malignancies. There were seven cases of malignant salivary gland tumors and 6 of squamous cell carcinoma. Two cases were high grade non-Hodgkin's lymphomas, and one was malignant odontogenic tumor. Of the 11 inflammatory or reactive lesions, 4 were found to be harboring Actinomyces. The rate of inadequate sampling was 6.7%. Histopathology reports on excised tissue were available for 10 cases only. Seven of nine cases with adequate cytology (77.88%) showed complete agreement with histology.     

Hydroxyurea affects cell morphology, cation transport, and red blood cell adhesion in cultured vascular endothelial cells

Hydroxyurea (HU) significantly increases fetal hemoglobin (Hb) production and concomitantly affects passive erythrocyte K transport and cell volume in patients homozygous for Hb S, thus decreasing disease severity. Red blood cells (RBCs) with Hb S display a greater adherence to vascular endothelial cells (VECs) than do Hb A cells, thus increasing the probability of vaso-occlusive crisis. The effect of HU on the structure and function of VECs is still unknown. In the present study, HU significantly changed, in a dose-dependent manner, the morphology and monovalent cation composition of cultured VECs after incubation in normal culture medium for up to 10 days in the absence and presence of 0.3 (therapeutic dose) and 3.0 (toxic dose) mmol/L HU. Treated cells showed significant morphologic changes such as an increase in apparent cell size and the formation of multinucleated giant cells. The protein content per dish decreased by 50% and 80% at 0.3 and 3.0 mmol/L HU, respectively, accompanied by an increase in cell Na (maximum, approximately 200%) and cell K (maximum, approximately 50%) contents at about days 4 to 6 and 8 to 10, respectively. In addition, HU decreased RBC adherence to VECs in experiments with 51Cr-loaded Hb A or Hb S RBCs. The HU-induced changes in VEC morphology, cation composition, and RBC adherence may be caused or accompanied by alterations in cell membrane permeability, transformation of endothelial cells, or decreased number/density of VEC adhesion molecules. Precise mechanisms of the HU effects warrant further investigation in light of the reported beneficial effects of HU in the treatment of sickle cell anemia.     

Monoclonal gammopathy and neuropathy

There is a known coexistence between polyneuropathy and monoclonal gammopathy (immunoglobulin M, immunoglobulin G, immunoglobulin A). Antibodies to several glycoconjugates of the peripheral nervous system have been found in 50-65% of patients with immunoglobulin M monoclonal proteins, and distinct clinical syndromes have been recognized. However, if no antibodies are found, a relationship between monoclonal protein and polyneuropathy is still debatable. Therapeutic intervention is generally directed at removing the autoantibodies by reducing the number of monoclonal B-cells.     

Polyneuropathy: diagnosis and management

A polyneuropathy is characterized by a symmetrical distribution of sensory or motor abnormalities, more pronounced distally than proximally and usually more evident in the lower than in the upper limbs. Polyneuropathies may be classified on the basis of (a) clinical picture: acute/subacute/chronic, sensory/motor/autonomous, axonal degeneration/segmental demyelinization, and (b) cause: metabolic disorder, deficiency, infection, auto(dys)immunity, hereditary and toxic/iatrogenic polyneuropathy, with idiopathic polyneuropathy as the remaining group. Damaged nerves may recover as the result of spontaneous remyelinization and axonal regeneration. Treatment is particularly successful in immunomediated neuropathies. Withdrawal is often successful in intoxications and suppletion in deficiencies. Even if no treatment is possible, the diagnosis is important: the patient can be taught to accept his disease and the prognosis can be determined, in connection with possible handicaps.     

The expression of voltage-dependent calcium channel beta subunits in human cerebellum

The beta subunits of voltage-dependent calcium channels, exert marked regulatory effects on the biophysical and pharmacological properties of this diverse group of ion channels. However, little is known about the comparative neuronal expression of the four classes of beta genes in the CNS. In the current investigation we have closely mapped the distribution of beta1, beta2, beta3 and beta4 subunits in the human cerebellum by both in situ messenger RNA hybridization and protein immunohistochemistry. To our knowledge, these studies represent the first experiments in any species in which the detailed localization of each beta protein has been comparatively mapped in a neuroanatomically-based investigation. The data indicate that all four classes of beta subunits are found in the cerebellum and suggest that in certain neuronal populations they may each be expressed within the same cell. Novel immunohistochemical results further exemplify that the beta voltage-dependent calcium channel subunits are regionally distributed in a highly specific manner and studies of Purkinje cells indicate that this may occur at the subcellular level. Preliminary indication of the subunit composition of certain native voltage-dependent calcium channels is suggested by the observation that the distribution of the beta3 subunit in the cerebellar cortex is identical to that of alpha(1E). Our cumulative data are consistent with the emerging view that different native alpha1/beta subunit associations occur in the CNS.     

Activation of the mas oncogene involves coupling to human alphoid sequences

We have shown previously that mouse NIH3T3 cells transfected with DNA from a human ovarian carcinoma were rendered tumourigenic by an activated mas oncogene in four independent transfection experiments. In all cases the 5'-noncoding region was rearranged in comparison to the original ovarian tumour DNA. We now report that in all four transfectants the newly acquired sequences consist of human centromeric alpha satellite repeat DNA. In at least three transfectants the alphoid DNA originates from the centromere of chromosome three. Analysis of the sequences of the recombination site in one transfectant revealed that a homologous sequence of five base pairs (CAGCA) is present in both parental strands, and might thus have contributed to the recombinational event. To establish a conclusive role for alphoid DNA in the activation of mas, we performed a co-transfection experiment in NIH3T3 cells with cloned alphoid DNA and the mas coding sequence. We show that the transfectants expressing a transformed phenotype contain amplified mas linked to alphoid DNA. NIH3T3 cells transfected with plasmids that contained alphoid sequences cloned directly upstream of the mas coding sequence, and injected into nude mice, gave rise to tumours with amplified mas sequences (7/7). In six of these tumours the alphoid sequences were amplified as well. Our data suggest a novel mechanism of oncogene activation: recombination with normal alphoid repeat DNA resulting in amplification of the oncogene.     

Measuring outcomes in nursing centers: otitis media as a sample case

While it is generally recognized that NPs offer affordable, quality health care, few studies have measured outcomes of clients who seek primary care services from NPs. This pilot study describes the outcomes of children with otitis media who received care from NPs employed in an academic nursing center. Outcome measurements included issues related to timing, level of analysis, and attribution. Parents of 27 children participated in a telephone survey consisting of seven questions relating to the care their children received from NPs and their recovery path. Although every respondent reported having a positive visit at the nursing center, concerns for NPs surfaced during the process of measuring outcomes. This study emphasizes the need for measuring outcomes in nursing clinics and demonstrates one way to measures client outcomes, revealing both general health care and specific nursing practice implications.