Reactivity of neoplastic cells of hairy cell leukemia with antisera to S-100 protein

Rabbit antibodies to bovine S-100 protein were tested by immunoperoxidase technics against fresh hairy cell leukemia (HCL) cells obtained from nine patients (peripheral blood in six and spleen in three), as well as lymphoblastoid cell lines derived from three patients with HCL. Peripheral mononuclear cells from three normal persons and two patients with chronic lymphocytic leukemia (CLL) and cells from two melanoma lines were used as controls. The melanoma cell lines, cell lines derived from patients with HCL, and fresh HCL cells displayed cytoplasmic and nuclear positivity after exposure to anti-S-100 protein sera. By contrast, normal peripheral blood lymphocytes and CLL cells were negative for S-100 protein. Additional studies were performed by immunoperoxidase technics on representative sections of formalin-fixed splenic tissues from eight patients who had splenectomies. The cause of splenectomy was HCL in three, traumatic rupture in one, CLL in one, Hodgkin's disease in one, and hypersplenism in two patients. Sections from all three HCL patients showed moderate to marked positivity with antisera to S-100 protein. These results strongly suggest the presence of S-100 protein in HCL cells.     

Clodronate inhibits the secretion of proinflammatory cytokines and NO by isolated microglial cells and reduces the number of proliferating glial cells in excitotoxically injured organotypic hippocampal slice cultures

Treatment of excitotoxically injured organotypic hippocampal slice cultures (OHSC) with clodronate is known to result in the inhibition of microglial activation. We hypothesized that this is due to direct effects of clodronate on microglial cells, and investigated microglial proliferation in OHSC, and cytokine and NO secretion in isolated microglial cells. N-methyl-D-aspartate (NMDA) lesioning of OHSC resulted in a massive increase in the number of proliferating, bromo-desoxy-uridine (BrdU)-labeled cells that was reduced to control levels after treatment with clodronate (0.1, 1, 10 microg/ml). Triple-labeling revealed that clodronate abrogated the proliferation of both glial fibrillary acidic protein (GFAP)-labeled astrocytes and Griffonia simplicifolia isolectin B4 (IB4)-labeled microglial cells. Furthermore, isolated microglial cells were treated with clodronate after stimulation with lipopolysaccharide (LPS) or macrophage colony stimulating factor (M-CSF). Clodronate (0.01, 0.1, 1 microg/ml) significantly down-regulated the LPS-stimulated microglial secretion of tumor necrosis factor (TNF)-alpha, Interleukin (IL)-1beta and NO, but not of IL-6. In contrast, clodronate significantly reduced the microglial IL-6-release induced by M-CSF, indicating different intracellular pathways. The number and morphology of isolated microglial cells did not change significantly after treatment with clodronate. In summary, the number of proliferating microglial cells and astrocytes after excitotoxic injury is reduced to control levels after treatment with clodronate. Furthermore, clodronate inhibits microglial secretion of proinflammatory cytokines and NO. Clodronate could therefore prove to be a useful tool in the investigation of interactions between damaged neurons and microglial cells.     

Evaluation of the effectiveness of anterior transposition of the inferior oblique muscle in dissociated vertical deviation with or without inferior oblique overaction

PURPOSE: Anterior transposition of the inferior oblique muscle (ATIO) has been reported to be an effective treatment for dissociated vertical deviation (DVD). In this study, we evaluated the effectiveness of this surgical procedure in patients with DVD alone and in those with DVD with concurrent overactive inferior oblique muscle and compared the results. METHODS: Nineteen eyes with DVD alone (group 1) and 15 eyes with DVD and concurrent inferior oblique overaction (group 2) underwent ATIO. The amount of DVD and inferior oblique overaction before and after the operation was measured and statistically analyzed. RESULTS: The average follow-up time for group 1 and group 2 was 9.4 and 9.0 months, respectively. Of a total of 34 eyes, 7 had DVD of more than 15 prism-diopters (pd) [four eyes from group 1 and three eyes from group 2] in which the residual DVD after surgery was more than 5 pd. However, in the 27 eyes with DVD of 15 pd or less (15 from group 1 and 12 from group 2), the residual DVD after the operation was less than 5 pd. (Fisher's exact test, P = .014). CONCLUSION: ATIO is an effective method for correcting DVD with and without inferior oblique overaction, especially in deviations of less than 15 pd.     

Combined intensive blood pressure and glycemic control does not produce an additive benefit on microvascular outcomes in type 2 diabetic patients

A reduction of either blood pressure or glycemia decreases some microvascular complications of type 2 diabetes, and we studied here their combined effects. In total, 4733 older adults with established type 2 diabetes and hypertension were randomly assigned to intensive (systolic blood pressure less than 120 mm Hg) or standard (systolic blood pressure less than 140 mm Hg) blood pressure control, and separately to intensive (HbA1c less than 0.060) or standard (HbA1c 0.070-0.079) glycemic control. Prespecified microvascular outcomes were a composite of renal failure and retinopathy and nine single outcomes. Proportional hazard regression models were used without correction for type I error due to multiple tests. During a mean follow-up of 4.7 years, the primary outcome occurred in 11.4% of intensive and 10.9% of standard blood pressure patients (hazard ratio 1.08), and in 11.1% of intensive and 11.2% of standard glycemia control patients. Intensive blood pressure control only reduced the incidence of microalbuminuria (hazard ratio 0.84), and intensive glycemic control reduced the incidence of macroalbuminuria and a few other microvascular outcomes. There was no interaction between blood pressure and glycemic control, and neither treatment prevented renal failure. Thus, in older patients with established type 2 diabetes and hypertension, intensive blood pressure control improved only 1 of 10 prespecified microvascular outcomes. None of the outcomes were significantly reduced by simultaneous intensive treatment of glycemia and blood pressure, signifying the lack of an additional beneficial effect from combined treatment.     

Insulin fibrillation and protein design: topological resistance of single-chain analogs to thermal degradation with application to a pump reservoir

Insulin is susceptible to thermal fibrillation, a misfolding process that leads to nonnative cross-β assembly analogous to pathological amyloid deposition. Pharmaceutical formulations are ordinarily protected from such degradation by sequestration of the susceptible monomer within native protein assemblies. With respect to the safety and efficacy of insulin pumps, however, this strategy imposes an intrinsic trade-off between pharmacokinetic goals (rapid absorption and clearance) and the requisite physical properties of a formulation (prolonged shelf life and stability within the reservoir). Available rapid-acting formulations are suboptimal in both respects; susceptibility to fibrillation is exacerbated even as absorption is delayed relative to the ideal specifications of a closed-loop system. To circumvent this molecular trade-off, we exploited structural models of insulin fibrils and amyloidogenic intermediates to define an alternative protective mechanism. Single-chain insulin (SCI) analogs were shown to be refractory to thermal fibrillation with maintenance of biological activity for more than 3 months under conditions that promote the rapid fibrillation and inactivation of insulin. The essential idea exploits an intrinsic incompatibility between SCI topology and the geometry of cross-β assembly. A peptide tether was thus interposed between the A- and B-chains whose length was (a) sufficiently long to provide the "play" needed for induced fit of the hormone on receptor binding and yet (b) sufficiently short to impose a topological barrier to fibrillation. Our findings suggest that ultrastable monomeric SCI analogs may be formulated without protective self-assembly and so permit simultaneous optimization of pharmacokinetics and reservoir life.     

A novel poly(2-mercaptobenzothiazole) coated magnetic nanoadsorbent derived from ZIF-8 for preconcentration/determination of palladium and silver

Herein, a novel poly(2-mercaptobenzothiazole) coated magnetic nanoadsorbent derived from zeolitic-imidazole framework-8 (ZIF-8) was synthesized and then employed for the extraction/preconcentration of trace amounts of palladium and silver in various real matrixes. In this way, magnetite was fabricated first, and then functionalized with tetraethyl orthosilicate. After that, the synthesized magnetite@silica was coated with the ZIF-8 to obtain magnetic ZIF-8. Afterward, the magnetic ZIF-8 was pyrolyzed under the protection of a nitrogen atmosphere to get a magnetic carbon nanoadsorbent. Finally, the magnetic carbon was functionalized with a conductive polymer (poly-2-mercaptobenzothiazole). Fabrication of the nanoadsorbent was affirmed with scanning and transmission electron microscopies, elemental analysis, X-ray diffraction, Fourier transform infrared spectroscopy, and vibrating sample magnetometry. The method is linear from 0.25 to 200 μg L⁻¹ for silver, and from 0.5 to 250 μg L⁻¹ for palladium. The detection limits are 0.07 and 0.15 μg L⁻¹ for Ag and Pd, respectively. The precision was evaluated at three concentration levels (1, 75, 200 μg L⁻¹, n = 5) and all the relative standard deviation (RSD) values were lower than 10.3%. In the end, the new method was utilized for the preconcentration/determination of trace amounts of palladium and silver in various real matrixes, satisfactorily (relative recovery: 86% to 104%; RSD%: 4.0–9.5%).

A novel poly(2-mercaptobenzothiazole) coated magnetic nanoadsorbent derived from zeolitic-imidazole framework-8 was synthesized and then employed for the extraction/preconcentration of trace amounts of palladium and silver in various real matrixes.     

Suppression of Lymphoblastoid Cell Line Proliferation by Antisera to HLA-DR and Other HLA Antigens

We have observed that certain antisera to HLA antigens suppress the in vitro proliferation of lymphoblastoid cell lines. An antiserum to HLA-B8 demonstrated suppressor activity associated with the presence of B8 antigen on the target LCLs; this activity was removed by absorption with platelets or with B8-positive LCLs. An antiserum to HLA-DRw7 suppressed proliferation of all six DRw7-positive LCLs and none of 12 DRw7-negative LCLs; this activity was removed by absorptions with three DRw7-positive LCLs, each representing a different HLA-D allele (i.e. Dw7, 10 and 11); it could not be removed by triple serial absorptions with platelets from multiple-donor pools. These experiments indicate that the suppressor activity of this serum is specifically associated with antibodies to DRw7. Our model of LCL growth suppression by HLA antisera is easily manipulated and provides a definitive tool for further characterizing HLA antigens and antisera in a complement-independent system.