Nanoparticle technology: addressing the fundamental roadblocks to protein biomarker discovery.

Clinically relevant biomarkers exist in blood and body fluids in extremely low concentrations, are masked by high abundance high molecular weight proteins, and often undergo degradation during collection and transport due to endogenous and exogenous proteinases. Nanoparticles composed of a N-isopropylacrylamide hydrogel core shell functionalized with internal affinity baits are a new technology that can address all of these critical analytical challenges for disease biomarker discovery and measurement. Core-shell, bait containing, nanoparticles can perform four functions in one step, in solution, in complex biologic fluids (e.g. blood or urine): a) molecular size sieving, b) complete exclusion of high abundance unwanted proteins, c) target analyte affinity sequestration, and d) complete protection of captured analytes from degradation. Targeted classes of protein analytes sequestered by the particles can be concentrated in small volumes to effectively amplify (up to 100 fold or greater depending on the starting sample volume) the sensitivity of mass spectrometry, western blotting, and immunoassays. The materials utilized for the manufacture of the particles are economical, stable overtime, and remain fully soluble in body fluids to achieve virtually 100 percent capture of all solution phase target proteins within a few minutes.

Ketoconazole inhibits organic osmolyte efflux and induces heat shock proteins in rat renal medulla.

Although ketoconazole (KC) is known to inhibit the cellular efflux of organic osmolytes in vitro, it is not known whether this effect can also be shown in vivo. Inhibition of osmolyte efflux by KC would impair osmotic adaptation and result in stress to the cells of the renal medulla when extracellular osmolality falls. Stress-inducible heat shock proteins (HSPs) may also participate in this response to osmotic stress. The aim of the present study was thus to establish whether KC inhibits organic osmolyte efflux from the cells of the renal medulla in vivo in response to a furosemide diuresis, and to establish whether HSPs are involved. A 20-minute furosemide infusion reduced urine osmolality and medullary urea content in control and KC-treated rats similarly. However, the efflux of methylamines (glycerophosphorylcholine, betaine) and polyols (myo-inositol, sorbitol) was attenuated in KC-treated rats while the efflux of amino acids was not significantly affected. Phosphorylation of HSP25 after the 20-minute furosemide diuresis was increased in KC rats. With continuing diuresis this returned to control levels after three hours. While short-term (up to 3 hr) diuresis did not alter the absolute amounts of HSPs in the renal medulla, long-term (24 or 48 hr) diuresis was associated with significantly increased amounts of HSP25 and HSP72 in KC-treated rats compared with control. These results suggest that KC inhibits the efflux of methylamines and polyols, thus impeding osmoadaptation of renal medullary cells during the onset of diuresis. This situation apparently increases the osmotic stress experienced by the cells of the renal medulla and provokes expression of HSP25 and HSP72.

Heat shock proteins HSP25, HSP60, HSP72, HSP73 in isoosmotic cortex and hyperosmotic medulla of rat kidney.

The distribution of heat shock proteins (HSP) HSP60, HSP73, HSP72 and HSP25 in the isoosmotic cortex and the hyperosmotic medulla of the rat kidney was investigated using Western blot analysis and immunohistochemistry. HSP73 was homogeneously distributed throughout the whole kidney. The level of HSP60 was high in the renal cortex and low in the medulla. HSP25 and HSP72 were present in large amounts in the medulla. Only low levels of HSP25 and almost undetectable amounts of HSP72 were found in the cortex. HSP25 exists in one nonphosphorylated and several phosphorylated isoforms. Western blot analysis preceded by isoelectric focussing showed that HSP25 predominates in its nonphosphorylated form in the outer medulla but in its phosphorylated form in cortex and inner medulla. Although this intrarenal distribution pattern was not changed during prolonged anaesthesia (thiobutabarbital sodium), a shift from the nonphosphorylated to the phosphorylated isoforms of HSP25 occurred in the medulla. The characteristic intrarenal distribution of the constitutively expressed HSPs (HSP73, HSP60, HSP25) may reflect different states of metabolic activity in the isoosmotic (cortex) and hyperosmotic (medulla) zones of the kidney. The high content of inducible HSP72 in the medulla most likely is a consequence of the osmotic stress imposed upon the cells by the high urea and salt concentrations in the hyperosmotic medullary environment.

Perioperative care of patients undergoing spinal stabilization with internal fixation (continuing education credit).

1. Low back pain affects approximately 80% of the adult population. There are between 200,000 and 500,000 spinal surgeries performed every year. 2. New spinal implant systems offer hope for persons with failed spinal surgery--as well as for those with spinal fractures, metastatic disease, and severe degenerative disorders. 3. Although complications can develop, the benefits of new spinal fixation systems far outweigh the problems.

[Somatostatin receptor scintigraphy in the primary diagnosis and follow-up care of gastrinoma]. / Somatostatinrezeptor-Szintigraphie in der Primärdiagnostik und Nachsorge bei Gastrinomen.

Somatostatin receptor scintigraphy (SRS) was performed in 14 patients (five men, nine women; mean age 51.5 [20-71] years) with Zollinger-Ellison syndrome (ZES), a gastrinoma proven in 7 and suspected on clinical or biochemical grounds in 7. The results were compared with those obtained by other methods (ultrasound, computed tomography, angiography). All 12 known tumour manifestations were demonstrated by SRS in seven patients with histologically confirmed gastrinoma. In four patients previously non-localized tumour was revealed by SRS, while in seven other patients the procedure led to modification of the treatment (primary tumour resection: n = 3, resection of metastases: n = 2, percutaneous radiation or chemoembolization: one each). These results suggest the following indications for SRS: (1) staging or re-staging in histologically proven gastrinoma and (2) search for primary tumour in clinically and biochemically suspected ZES.

Impact of the 1993 revision of the AIDS case definition on the prevalence of AIDS in a clinical setting.

OBJECTIVE: To determine the impact of the 1993 revision of the Centers for Disease Control and Prevention (CDC) AIDS surveillance case definition on the prevalence of AIDS. DESIGN: Review of prospectively collected baseline clinical and demographic data on HIV-infected patients presenting for care between December 1988 and May 1991. SETTING: The HIV Clinic of the Johns Hopkins Hospital, an urban, primary care institution. MAIN OUTCOME MEASURE: Diagnosis of AIDS by the 1987 (specific indicator diseases) or the 1993 (indicator diseases, pulmonary tuberculosis, recurrent bacterial pneumonia, cervical carcinoma, or CD4 lymphocyte count < 200 x 10(6)/l) CDC case definition. RESULTS: Of 955 patients evaluated, 122 (13%) had AIDS by the 1987 case definition at presentation. An additional 126 (13%) met the 1993 but not the 1987 case definition. Patients meeting only the 1993 case definition were more likely to be female [28 versus 14%; odds ratio (OR), 2.4; 95% confidence interval (CI), 1.2-3.0; P = 0.01] and intravenous drug users (40 versus 26%; OR, 2.0; 95% CI, 1.1-3.3; P = 0.02) than patients meeting the 1987 case definition. Fifty-five per cent of patients meeting only the 1993 case definition were asymptomatic, and 7% (nine patients) had new indicator diseases but CD4 counts > 200 x 10(6)/l. Median time to progression from a diagnosis of AIDS by the 1993 case definition to diagnosis by the 1987 case definition was 435 days. Patients with AIDS by the 1987 case definition had a median survival of 594 days from presentation (2-year survival, 42%), while median survival time for patients with AIDS by the 1993 case definition only was 947 days (2-year survival, 60%; P < 0.005). CONCLUSIONS: The proposed 1993 revision of the AIDS surveillance case definition would double the number of prevalent AIDS cases, with significant increases in the proportion of cases who are female, intravenous drug users, or asymptomatic. Survival of patients meeting the 1993 case definition is significantly longer than that of patients meeting the 1987 case definition. The new AIDS case definition will have a major impact both on AIDS surveillance and on medical and social service programs that use diagnosis of AIDS as a criterion for eligibility for services.

Splash basin contamination in orthopaedic surgery.

Samples of splash basin fluid were cultured at the end of 78 randomly selected orthopedic operations. Fifty-eight (74%) of the specimens were positive on culture. Staphylococcus epidermidis was the prevalent organism. Thirty-four (59%) of the positive cultures grew multiple organisms. Seven (12%) grew more than 100 colonies per 100 ml specimen. This study demonstrates that splash basin fluid is frequently contaminated and may be a source of wound contamination during orthopedic surgery. Implants should not be placed in the splash basin, and instruments placed in it should not be returned to the operative wound.