Design and Synthesis of Soluble and Cell-Permeable PI3Kδ Inhibitors for Long-Acting Inhaled Administration.

PI3Kδ is a lipid kinase that is believed to be important in the migration and activation of cells of the immune system. Inhibition is hypothesized to provide a powerful yet selective immunomodulatory effect that may be beneficial for the treatment of conditions such as asthma or rheumatoid arthritis. In this work, we describe the identification of inhibitors based on a thiazolopyridone core structure and their subsequent optimization for inhalation. The initially identified compound (13) had good potency and isoform selectivity but was not suitable for inhalation. Addition of basic substituents to a region of the molecule pointing to solvent was tolerated (enzyme inhibition pIC50 > 9), and by careful manipulation of the pKa and lipophilicity, we were able to discover compounds (20b, 20f) with good lung retention and cell potency that could be taken forward to in vivo studies where significant target engagement could be demonstrated.

United States Army Research Institute of Environmental Medicine: Warfighter research focusing on the past 25 years.

The United States Army Research Institute of Environmental Medicine (USARIEM) celebrated its 50th anniversary on July 1, 2011. This article reviews its history, evolution, and transition of its research programs as well as its scientific and military accomplishments, emphasizing the past 25 yr. During the 1990s, USARIEM published a series of pocket guides providing guidance for sustaining Warfighter health and performance in Southwest Asia, Somalia, the former Republic of Yugoslavia, Rwanda, and Haiti. Issues identified during Operation Desert Storm elicited research that improved nutritional guidelines for protracted desert operations; safer use of nuclear, chemical, and biological protective clothing; equipment, development, and fielding of efficient microclimate cooling systems; and effective evaluation of pharmaceuticals to protect soldiers from chemical and biological threats. During the first decade of the 21st century, USARIEM and the Department of the Army published official medical/performance doctrines for operations in the heat and cold and at high altitude. The current Global War on Terrorism focused research to improve doctrines for hot, cold, and high-altitude operations, reduce musculoskeletal training injuries, provide improved field nutrition, more efficient planning for operational water requirements, and improve both military clothing and materiel. This article also describes the critically important interactions and communications between USARIEM and deployed units and the benefits to Warfighters from this association. This report presents USARIEM's unique and world-class facilities, organizational changes, scientific and support personnel, and major research accomplishments, including the publication of 2,200 scientific papers over the past 25 yr.

Discovery of 8-azabicyclo[3.2.1]octan-3-yloxy-benzamides as selective antagonists of the kappa opioid receptor. Part 1.

Initial high throughput screening efforts identified highly potent and selective kappa opioid receptor antagonist 3 (κ IC(50)=77 nM; µ:κ and δ:κ IC(50) ratios>400) which lacked CNS exposure in vivo. Modification of this scaffold resulted in development of a series of 8-azabicyclo[3.2.1]octan-3-yloxy-benzamides showing potent and selectivity κ antagonism as well as good brain exposure. Analog 6c (κ IC(50)=20 nM; µ:κ=36, δ:κ=415) was also shown to reverse κ-agonist induced rat diuresis in vivo.

Modeling physiological responses to military scenarios: initial core temperature and downhill work.

INTRODUCTION: Previous field studies suggested that a thermoregulatory model developed by the U.S. Army Research Institute of Environmental Medicine (USARIEM) needed an adjustment of initial core temperature (Tcr) for individual variation and a metabolic (M) correction during downhill movements. This study evaluated the updated version of the model incorporating these new features using a dataset collected during U.S. Marine Corps marksmanship training at Quantico, VA. METHODS: Individual anthropometrics, physiological, and environmental time series data were obtained from five Marine men. The study focused on the marksmanship training for approximately 2 h, then 30-min marching including uphill and downhill movements in a moderately hot environment (air temperature: approximately 30 degrees C; dew point: approximately 21 degrees C). The predicted and observed heart rate (HR) and Tcr measurements were compared by root mean square deviations (RMSD). RESULTS: Overall, the current model improved predictions of physiological measures (HR RMSD = 23 bpm, Tcr RMSD = 0.46 degrees C), particularly for marching in the heat (HR RMSD = 21 bpm, Tcr RMSD = 0.32 degrees C). The model under-predicted both HR and Tcr during marksmanship training, indicating that a greater solar effect or non-thermal factors may have required higher M rates during these periods. CONCLUSIONS: Updated features of the model significantly improved physiological predictions. However, accurate M estimates are required for slow movements of subjects under heat stress, such as movements on the firing range. Such improvement should result in more accurate simulations of physiological status and better risk assessment, thereby reducing heat injuries and improving performance of deployed military personnel.

Sibling donor cord blood banking for children with sickle cell disease.

Although hematopoietic stem cell transplantation has curative potential for selected patients with sickle cell disease (SCD), most patients who are eligible for transplantation do not have a suitable donor. Cord blood (CB) from a sibling could provide an alternative stem cell source that, while not as well established as marrow, may offer certain advantages for selected families. These potential advantages include low risk to the infant donor, the possibility that mismatched CB units from sibling donors may be acceptable for transplantation, prompt availability of a stored CB unit for transplant, and decreased risk of clinically significant graft-versus-host disease. When families with SCD (or other transplant-treatable condition) conceive a sibling, no comprehensive research resource exists to assist the family in collecting the new infant's CB. With support from the National Heart Lung and Blood Institute, we are developing a noncommercial research-based CB Banking Program specifically for medically indicated sibling donations. In preliminary experience, we have collected CB from 52 SCD families across 19 states. Of these, 2 CB units have thus far been used for transplantation and 9 others are HLA-identical. We conclude that a CB bank focusing on sibling-donations may be feasible, but further study is required to determine whether such a bank can collect CB units of sufficient quantity and quality to support controlled trials of sibling CB transplantation. Families with a specific medical need, such as those already caring for a child with SCD, should consider collecting sibling CB as part of comprehensive care if the opportunity becomes available.

Development of nuclear transfer and parthenogenetic rabbit embryos activated with inositol 1,4,5-trisphosphate.

The present study was carried out to evaluate the effects of different activation protocols, enucleation methods, and culture media on the development of parthenogenetic and nuclear transfer (NT) rabbit embryos. Electroporation of 25 mM inositol 1,4, 5-trisphosphate (IP3) in calcium- and magnesium-free PBS immediately induced a single intracellular calcium transient in 6 out of 14 metaphase II-stage rabbit oocytes evaluated during a 10-min recording period. The percentage of oocytes treated with IP3 followed by 6-dimethylaminopurine (IP3 + DMAP) that cleaved (83.9%) and reached the blastocyst stage (50%) was significantly higher (p < 0.05) than those activated with multiple pulses (61.6% and 30.1%, respectively) or treated with ionomycin + DMAP (52.9% and 5.7%, respectively). Development of IP3 + DMAP-activated rabbit oocytes and in vivo-fertilized zygotes in different culture media was studied. Development of activated oocytes to the blastocyst stage in Earle's balanced salt solution (EBSS) supplemented with MEM nonessential amino acids, basal medium Eagle amino acids, 1 mM L-glutamine, 0.4 mM sodium pyruvate, and 10% fetal bovine serum (FBS) (EBSS-complete) (40.6%) was significantly higher (p < 0.05) than those that developed in either Dulbecco's Modified Eagle's medium (DMEM)/RPMI + 10% FBS (15.5%) or CR1aa + 10% FBS (4%) medium. In addition, 100% of in vivo-fertilized rabbit zygotes developed to the blastocyst stage in EBSS-complete. A third set of experiments was carried out to study the efficiency of blind versus stained (Hoechst 33342) enucleation of oocytes. Twenty-nine of 48 blind enucleated and IP3 + DMAP-activated oocytes cleaved (60.4%), and 15 (31.2%) subsequently reached the blastocyst stage, whereas 9 of 52 oocytes enucleated using epifluorescence (17.3%) cleaved, and none of these reached the blastocyst stage. When the above parameters that yielded the highest blastocysts were combined in an NT experiment using adult rabbit fibroblast nuclei, 72.2% (39 of 54) of the fused nuclear transplant embryos cleaved and 29.6% (16 of 54) reached the blastocyst stage.

Chiropractic management of primary nocturnal enuresis.

OBJECTIVE: To evaluate chiropractic management of primary nocturnal enuresis in children. DESIGN: A controlled clinical trial for 10 wk preceded by and followed by a 2-wk nontreatment period. SETTING: Chiropractic clinic of the Palmer Institute of Graduate Studies and Research. PARTICIPANTS: Forty-six nocturnal enuretic children (31 treatment and 15 control group), from a group of 57 children initially included in the study, participated in the trial. INTERVENTION: High velocity, short lever adjustments of the spine consistent with the Palmer Package Techniques; or a sham adjustment using an Activator at a nontension setting administered to the examiner's underlying contact point. Two 5th-year chiropractic students under the supervision of two clinic faculty performed the adjustments. MAIN OUTCOME MEASURES: Frequency of wet nights. RESULTS: The post-treatment mean wet night frequency of 7.6 nights/2 wk for the treatment group was significantly less than its baseline mean wet night frequency of 9.1 nights/2 wk (p = 0.05). For the control group, there was practically no change (12.1 to 12.2 nights/2 wk) in the mean wet night frequency from the baseline to the post-treatment. The mean pre- to post-treatment change in the wet night frequency for the treatment group compared with the control group did not reach statistical significance (p = 0.067). Twenty-five percent of the treatment-group children had 50% or more reduction in the wet night frequency from baseline to post-treatment while none among the control group had such reduction. CONCLUSIONS: Results of the present study strongly suggest the effectiveness of chiropractic treatment for primary nocturnal enuresis. A larger study of longer duration with a 6-month follow-up is therefore warranted.

Ethiodized oil emulsion enhanced computerized tomography in the preoperative assessment of metastases to the liver from the colon and rectum.

Selective reticuloendothelial (RE) cell uptake of ethiodized oil emulsion 13 (EOE-13), an emulsion of Ethiodol (ethiodized oil) roentgenographic contrast material in a phosphate buffer, permits detection of small metastatic lesions of the liver and spleen through enhancement of roentgenographic density differences on computerized tomography (CT) between tissues containing and not containing RE cells. To determine the efficacy of this contrast material in the assessment of patients with metastatic disease of the liver, routine CT and emulsion enhanced tomography (EOE) were performed in a series of 15 patients prior to surgical exploration for treatment of carcinoma of the colon and rectum. All patients were suspected of harboring hepatic metastasis on the basis of clinical examination, liver function tests or radionuclide scans. EOE consistently demonstrated the nature and location of hepatic defects. Surgical exploration failed to locate one metastasis that was judged to be real because of progressive enlargement on EOE and CT over a period of two years. CT scans detected metastases in three patients subsequently shown to have normal livers and failed to detect disease in one patient subsequently shown to have metastases. EOE contrast material provides a more sensitive and accurate picture of metastatic liver involvement from carcinoma of the colon and rectum than is available on routine CT. The information provided by the results of this test can be useful in preoperative planning when treatment of disease of the liver is considerable feasible.

Comparison of azlocillin and ticarcillin in the treatment of urinary tract infection.

This prospective, controlled, randomized double blind study compared the safety and effectiveness of intravenous azlocillin at a dosage of 6 g/day in three divided doses with that of intravenous ticarcillin at 8 g/day in four divided doses for the treatment of urinary tract infections in 35 patients. The clinical and bacteriological responses among the 26 courses evaluable for drug effectiveness (13 in each group) appeared to be somewhat more favourable in the azlocillin treatment group (92.3%) than the ticarcillin group (69.2%). However, statistically, both treatment groups did not differ in a significant manner in regard to effectiveness or adverse reactions. Of the patients who were followed for relapses for three-four weeks after treatment, 66.7% of the azlocillin group and 45.5% of the ticarcillin group remained free of the original infecting organism. Local and systemic tolerance for both drugs was excellent.

Anaesthesia for day-care surgery: a symposium (III). Anaesthesia for adult surgical out-patients.

This discussion is based on the experience of the Phoenix Surgicenter, where over 60,000 patients have been anaesthetized since 1970. Patients accepted for out-patient surgery are ASA Status I or II, although status III patients may be included if their co-existing disability is under excellent control. Eighty-five per cent of adult patients receive general anaesthesia. A wide variety of local and regional anaesthetic techniques may be used. Efforts during recovery are directed towards preparing the patient for discharge in a "home ready" condition for safe handling by attending relatives. The common complications have been postoperative nausea or emesis and hypotension. The hospital transfer rate has been 0.2 per cent.

Immunologic function in humans before and after hyperthermia and chemotherapy for disseminated malignancy.

Multiple immunologic parameters were studied in three patients prior to and after hyperthermia treatment for disseminated malignancy. Two patients had malignant melanoma and received chemotherapy during the hyperthermia treatment. One had adenocarcinoma of the stomach and received no concomitant chemotherapy. Rapid rosettes as a measure of thymus-derived lymphocytes (T-lymphocytes) were found to increase significantly after therapy (P less than 0.05) both in percentage and absolute numbers. There was no change in the numbers or percentages of other markers for T-lymphocytes or bone marrow-derived B-lymphocytes. Complement profiles revealed a significant decrease in C3 (P less than 0.005) after hyperthermia but no change in levels of other components of the alternate pathway. Antibody-dependent lymphocyte-mediated cytotoxicity and polymorphonuclear cell-mediated antibody-dependent cytotoxicity were also depressed after hyperthermia. No change was observed in immunoglobulin levels with hyperthermia therapy. Results indicated that hyperthermia may favorably alter the immune balance between tumor and host in selected instances.

Role of immunoglobulins in protection against Shigella-induced keratoconjunctives.

Various immunoglobulin preparations were tested for their ability to protect guinea pig eyes from infection by a virulent strain of Shigella flexneri. Secretory immunoglobulin A was effective in delaying or preventing keratoconjunctivitis in eight guinea pigs when it was used to precoat the organism, and was also placed in the eye with the inoculum. Neither immunoglobulin G nor immunoglobulin M gave any protection when used in the same way. Protection by secretory immunoglobulin A appeared to be related to the antishigella antibody content of the immunoglobulin since a low-titered preparation gave less protection than a higher-titered one.