ADAM10 and Notch1 on murine dendritic cells control the development of type 2 immunity and IgE production.

BACKGROUND: Allergy and allergic asthma are significant health burdens in developed countries and are increasing in prevalence. Dendritic cells (DCs) initiate immune responses to common aeroallergens, and ADAM10 has been demonstrated to be important for the development of adaptive responses. This study's objective was to understand the role of ADAM10 on DCs in the development of allergic and anaphylactic responses. METHODS: In this study, we used mouse models of allergic airway inflammation (house dust mice and Alternaria alternata) and OVA-induced models of active anaphylaxis to determine the DC-specific function of ADAM10 and Notch signaling. To examine TH 1 and TH 17 immunity infection with Anaplasma phagocytophilum and Citrobacter rodentium respectively, were used. RESULTS: Mice, which have ADAM10 deleted from DCs, have dramatic reductions in IgE production and do not develop significant TH 2 immune responses. Further, ADAM10DC-/- mice are resistant to IgE-mediated anaphylaxis. This response is selective for TH 2 immunity as TH 1 and TH 17 immunity is largely unaffected. Notch1, a known ADAM10 substrate, when knocked out of DCs (Notch1DC-/- ) demonstrated a similar reduction in anaphylaxis and IgE. Without ADAM10 and Notch1 signaling, DCs were unable to make cytokines that stimulate TH 2 cells and cytokines. Anaphylaxis and allergic lung inflammation were restored in ADAM10DC-/- with the overexpression of the Notch1-intracellular domain, confirming the role of Notch signaling. CONCLUSIONS: Targeting ADAM10 and Notch1 on DCs represent a novel strategy for modulating TH 2 immune responses and IgE production.

Multiple Bone Lesions in an 8-Month-Old Child Presenting with Pathologic Fracture: A Rare Case of Solely Osseous Multicentric Infantile Myofibromatosis.

CASE: An otherwise healthy 8-month-old boy presented with a pathologic fracture of the distal aspect of the radius. Further work-up demonstrated widespread osseous lesions of the axial and the appendicular skeleton with no soft-tissue or visceral involvement. CONCLUSION: Infantile myofibromatosis has a spectrum of severity that demands a careful and complete work-up. In rare cases such as the present one, it can manifest as multiple osseous lesions. The patient in the present case was managed conservatively, with no morbidity demonstrated at 1 year of follow-up.

Amplification of pfmdr 1 associated with mefloquine and halofantrine resistance in Plasmodium falciparum from Thailand.

Drug resistance in Plasmodium falciparum is an expanding problem in most endemic areas. Recent studies have suggested the potential involvement of genes in the MDR gene family in resistance to quinoline-containing compounds in P. falciparum. In this study a molecular analysis of pfmdr 1 in recent isolates from Thailand was done (1) to further examine the role of pfmdr 1 in drug-resistant isolates and (2) to examine the reported association of pfmdr 1 intragenic alleles and chloroquine resistance. Most of the isolates (10 of 11) were resistant to all compounds tested. Analysis of pfmdr 1 revealed an apparent association between increased gene copy number and increased level of expression of pfmdr 1 and decreased susceptibility to mefloquine and halofantrine. Sequence analysis of pfmdr 1 in these isolates revealed no association of intragenic alleles with chloroquine resistance.

Serial analysis of gene expression (SAGE) in Plasmodium falciparum: application of the technique to A-T rich genomes.

The advent of high-throughput methods for the analysis of global gene expression, together with the Malaria Genome Project open up new opportunities for furthering our understanding of the fundamental biology and virulence of the malaria parasite. Serial analysis of gene expression (SAGE) is particularly well suited for malarial systems, as the genomes of Plasmodium species remain to be fully annotated. By simultaneously and quantitatively analyzing mRNA transcript profiles from a given cell population, SAGE allows for the discovery of new genes. In this study, one reports the successful application of SAGE in Plasmodium falciparum, 3D7 strain parasites, from which a preliminary library of 6880 tags corresponding to 4146 different genes was generated. It was demonstrated that P. falciparum is amenable to this technique, despite the remarkably high A-T content of its genome. SAGE tags as short as 10 nucleotides were sufficient to uniquely identify parasite transcripts from both nuclear and mitochondrial genomes. Moreover, the skewed A-T content of parasite sequence did not preclude the use of enzymes that are crucial for generating representative SAGE libraries. Finally, a few modifications to DNA extraction and cloning steps of the SAGE protocol proved useful for circumventing specific problems presented by A-T rich genomes.

Characterization of a Leishmania isolate from the rodent host Neotoma micropus collected in Texas and comparison with human isolates.

We report the biological and biochemical parameters of Leishmania parasites (MNEO/US/90/WR972) isolated from a rodent host, Neotoma micropus, collected in Texas. Footpad inoculations of WR972 promastigotes into BALB/c mice and Syrian hamsters resulted in ulcerating lesions six and eight weeks post-inoculation, respectively. Using monoclonal antibody-stained touch preparations, amastigotes were found in the liver of both laboratory hosts. Infection of J774 macrophages with WR972 promastigotes supported the growth of amastigotes for 12 days at 35 degrees C. The WR972 parasite was identified by enzyme electrophoresis as L. mexicana. Isozyme comparison of WR972 with 42 L. mexicana isolates (from humans and rodents) from four different endemic areas, including Texas, suggest that these parasite populations are identical for approximately 97% of their genetic loci. Pulse field gel electrophoresis (PFGE) of WR972 resolved 18 chromosomes with a size range of 300- greater than 2,000 kb. The karyotype strongly resembles that of two other Texas L. mexicana isolates from humans. Taken together, the PFGE, hybridization, and isoenzyme data suggest that the wood rat isolate (WR972) is identical to parasites from human cutaneous lesions isolated in Texas and Central America. In addition, the biological characteristics of WR972, its infectivity of BALB/c mice and the Syrian hamster, and the potential of the isolate to infect, transform, and divide in J774 macrophages indicate that WR972 will be pathogenic in humans if transmission occurs. Health care providers should consider this possibility when studying the epidemiology and control of cutaneous leishmaniasis in Texas.

Characteristics of multidrug resistance in Plasmodium and Leishmania: detection of P-glycoprotein-like components.

Multidrug-resistance (MDR) in neoplastic cells is frequently characterized by the overexpression of P-glycoprotein (PGP), a 170 kDa transmembrane glycoprotein that binds multiple cytotoxic drugs as well as calcium channel antagonists. Chloroquine resistance in Plasmodium falciparum appears to be analogous to MDR in neoplastic cells, where the induction of resistance with one drug confers resistance to other structurally and functionally unrelated drugs. To test the hypothesis that chloroquine resistance in P. falciparum and antimony resistance in Leishmania is mediated by a similar mechanism of MDR in mammalian neoplastic cells, a PGP-specific monoclonal antibody (C219) was used to determine the presence of PGP genes in resistant and sensitive Plasmodium and Leishmania parasites by indirect immunofluorescence assays and Western blotting procedures. These PGP-like components were detected in both drug-sensitive and -resistant Plasmodium and Leishmania cells. A 40-42 kDa component was observed to be greater in a chloroquine-resistant P. berghei (C line) than in a chloroquine-susceptible P line. Differences observed between Pentostam-resistant and -sensitive Leishmania promastigote clones and isolates included the increased expression of 96-106 and 23-25 kDa peptides in drug-resistant L. enrietti, and increased amounts of two different peptides in two drug-resistant L. panamensis clones (i.e., 96-106 and 43-45 kDa in WR-746-CL4, and 53 and 23-25 kDa in kDa) in amastigotes as in MDR KB carcinoma cells (KB-V1). Comparative indirect immunofluorescent studies suggested that a correlation existed between the degree of antimony susceptibility and the concentration of the moiety recognized by C219 in two L. panamensis clones. Binding of the C219 monoclonal antibody to the PGP-like component of Leishmania was blocked by Pentostam, while the binding of C219 to multiple-drug resistant KB-V1 PGP was not inhibited by Pentostam, regardless of the PGP concentration. This suggests some degree of specificity in the binding of Pentostam to the Leishmania PGP-like components. In addition, these studies have demonstrated that drug-sensitive Leishmania accumulate two to five times more 125Sb-Pentostam than resistant clones.

Detection of ptc in archival formalin-fixed, paraffin-embedded tissues. Comparison of radiolabeled DNA hybridization and direct incorporation of digoxigenin-11-dUTP into RT-PCR products.

Archival pathological specimens are a source of RNA and DNA for clinical surveillance or retrospective studies. We employed a modification of the acid guanidium thiocyanate-phenol-chloroform extraction method for the recovery of total RNA from formalin-fixed, paraffin-embedded neoplastic thyroid tissue. The extracted RNA was used for reverse transcription of ptc and subsequent amplification of the complementary DNA (cDNA) by the reverse transcription-polymerase chain reaction (RT-PCR). In lieu of 32P-labeled DNA for hybridization studies, we supplemented the nucleotide pool in the amplification reaction with a modified pyrimidine, digoxigenin-11-dUTP. Digoxigenin-11-dUTP was incorporated directly into the PCR product, eliminating the need for hybridization, posthybridization washes, and prolonged autoradiography. These products were resolved by electrophoresis on agarose gels, Southern blotted to nylon membranes, and rapidly detected by chemiluminescence. This nonradioisotopic method has expedited and reduced the cost for molecular investigations with archival pathological specimens by providing equal sensitivity to or greater sensitivity than that of DNA-labeled radionuclides without the associated biological hazards.

Preservation of nucleic acids for polymerase chain reaction after prolonged storage at room temperature.

A guanidinium isothiocyanate (GITC) lysis solution was evaluated for its efficacy in preserving nucleic acids for subsequent analysis after prolonged storage at room temperature. Aliquots of thyroid crude cell lysates were stored at 22 degrees C for 8 weeks in the GITC solution. Crude cell lysates stored for 2 weeks in the GITC solution consistently provided adequate RNA and DNA for reverse transcriptase-polymerase chain reaction (RT-PCR) and PCR, respectively, of beta 2-microglobulin, thyroid stimulating hormone receptor (TSHR), and thyroglobulin. Interestingly, the thyroglobulin and TSHR messages from thyroid fine needle aspirate samples were detected by RT-PCR only when stored at room temperature for less than 1 week, whereas the RT-PCR product for beta 2-microglobulin was detectable in 95% of the samples at 3 months. In addition, thyroglobulin DNA was amplified by PCR in nearly all samples stored at 22 degrees C for 2 months. These data suggest that GITC solutions can be used to preserve nucleic acids in most circumstance until transported to laboratories equipped and staffed with appropriate resources.

Double-blind, randomized comparison of olanzapine versus fluphenazine in the long-term treatment of schizophrenia.

This study was undertaken to evaluate the efficacy and safety of olanzapine compared with fluphenazine in the treatment of patients who met the Diagnostic and Statistical Manual, fourth edition (DSM-IV) diagnostic criteria for schizophrenia or schizoaffective disorder. This was a long-term (22-week), randomized, double-blind, parallel clinical trial. Sixty patients (mean age, 35.4 years) were randomly assigned to either olanzapine (n=30) or fluphenazine (n=30). They received treatment at three centers in Croatia during a 22-week study period and were assessed weekly for the first 6 weeks and monthly thereafter. Efficacy was measured using the Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Rating Scale (PANSS) and the Clinical Global Impression (CGI) Severity and Improvement scores. The Hillside Akathisia Scale (HAS), Simpson-Angus Scale (SAS), Abnormal Involuntary Movement Scale (AIMS), vital signs, laboratory tests, and treatment-emergent adverse events were assessed to evaluate safety. The olanzapine group showed significantly greater mean decreases from baseline to endpoint for BPRS total (-25.8 vs. -16.5, P=.035), PANSS total (-45.7 vs. -29.5, P=.037), PANSS positive (-13.0 vs. -7.9, P=.034), and CGI Severity (-2.2 vs. -1.3, P=.031) scores. The olanzapine group showed greater mean decreases on all measures of extrapyramidal symptoms, significantly so for the SAS (-2.1 vs. 1.9, P=.004) and HAS (-3.4 vs. 2.6, P=.028). Patients in the fluphenazine group experienced a higher incidence of treatment-emergent adverse events (76.7% vs. 50.0%, P=.032). Weight gain was the most frequently reported adverse event in the olanzapine group (16.7% vs. 0.0%, P=.020). Akathisia (30.0% vs. 10.0%, P=.053) and insomnia (20.0% vs. 0.0%, P=.010) appeared most frequent in the fluphenazine group. Daily use of anticholinergics and benzodiazepines were both significantly greater for the fluphenazine group (P=.003 and.04, respectively). No significant changes were observed in vital signs, ECG, or clinical chemistry. The study indicates that olanzapine has advantages in both efficacy and safety compared to fluphenazine; however, the small sample size limits our ability to draw definitive conclusions.

Olanzapine versus clozapine in treatment-resistant or treatment-intolerant schizophrenia.

Clozapine has been the gold standard for treatment of patients with refractory schizophrenia but is associated with serious safety liabilities. This has prompted the search for therapeutic alternatives for treatment-resistant schizophrenia. The objective of this study was to compare the efficacy and safety of olanzapine versus clozapine in schizophrenic patients who failed to respond adequately to antipsychotic medication or who experienced intolerable adverse effects associated with the medication. This 18-week, randomized, double-blind, parallel study compared treatment with either olanzapine (5-25 mg/day, n=75) or clozapine (100-500 mg/day, n=72) in patients with schizophrenia who were nonresponsive to, or intolerant of, standard acceptable antipsychotic therapy. At the 18-week endpoint, no statistically significant differences were found between olanzapine and clozapine in any efficacy measure used: Positive and Negative Syndrome Scale (PANSS) total, positive, negative, or general psychopathology or Clinical Global Impression severity (CGI-S). Response rates based on the criteria of Kane et al. [Arch. Gen. Psychiatry 45 (1988) 789] were also not significantly different between olanzapine-treated (57.9%) and clozapine-treated patients (60.8%). There were no significant differences in measurements of extrapyramidal symptoms or electrocardiography, and no clinically and statistically significant changes were seen in vital signs or laboratory measures in either group. Both treatments were well tolerated. Olanzapine demonstrated similar efficacy to clozapine in patients who had failed previous treatment because of lack of efficacy (treatment resistance) or intolerable side effects (treatment intolerance). Olanzapine therefore presents a safe alternative in the treatment of refractory schizophrenia.

Innate resistance to new antimalarial drugs in Plasmodium falciparum from Nigeria.

The rapid dissemination of chloroquine-resistant Plasmodium falciparum in West Africa has been well documented and represents a significant health threat to autochthonous populations. The methodical development of alternative chemotherapeutic agents demands that dispensing new antimalarial drugs (mefloquine, halofantrine, and artemisinine [qinghaosu]) be closely monitored in order to protect their clinical utility. Indeed, mefloquine-resistant strains of P. falciparum have been reported. We present data from experiments in vitro on the innate resistance of P. falciparum isolates to mefloquine as well as a disturbing observation of transient resistance to artemisinine. The implications for the extended efficacy of these new antimalarial drugs are addressed.

Load-carrying effects on the adult beagle tibia.

The response of the skeleton to carrying extra weight while exercising was studied in 30 young adult male beagles. Following 34 wk of a caged control period, 18 dogs carried lead-weighted jackets for 1 h and 15 min, 5 d/wk for 2 yr, on a carousel-type treadmill at 3.3 kph. During the next 23 wk, the load carried was gradually increased to 130% of the animal's body weight, and then maintained for 48 wk. The other 12 dogs served as controls. The dogs remained healthy and responded to exercise showing decreased skinfolds and body weight. All dogs showed increases in both tibial mineral content and width with time; but, the exercise group increased significantly more in mineral content than the controls. It is concluded that the gradual increase in bone mineral and width of the tibia is normal in the epiphyseal-closed skeleton of a young adult beagle. Also important is the fact that the intervention of exercise with added weight will increase tibial mineral content above these normal levels.

Cross-sectional thigh components: computerized tomographic assessment.

The purpose of this study was to examine the efficacy of the Picker Synerview Fourth Generation CT Scanner in assessing cross-sectional areas of muscle, bone, and subcutaneous fat of the thigh. A secondary purpose was to resolve individual muscle bellies using the CT scanner and to determine the effects of body composition in isolating those muscles. Both limbs of one cadaver were used to compare actual cross-sectional areas with the CT images. Each limb was dissected and photographed. Accuracy of repeated measurements was assessed using human volunteers. Absolute and percent error between the CT measurements and the measurements of the cross-sectional areas differed most at the bone. No differences were found between repeat scans. CT images and photographic measurements of individual muscles were generally quite similar, as assessed by absolute and percent error. The gracilis and sartorious were reasonably well defined in those individuals with more than 8% body fat. The results are discussed in terms of possible applications of the CT scanner in sports medicine research.

Influence of age and sex on the strength of bone-ligament junctions in knee joints of rats.

The bone-ligament junction strength of femur-medial collateral ligament-tibia complexes in rats was measured in situ at various ages during a two-year period. Male rats had a higher junction strength than female rats, a difference that became apparent when the animals were sixty days old and in male but not female animals subsequently paralleled the changes in body weight. However, on a bodyweight basis, the junctions were stronger in female than in male rats and this sex difference was evident at fifteen days old and persisted thereafter. Regression analysis between body weight and junction strength indicated that female rats had a significantly higher slope than males, which suggested that the sex differences were due to a hormonal factor or factors. Other measurements showed that elastic stiffness, failure energy, and collagen concentration in the ligament increased, whereas the water content of the ligament decreased with age. Most of these changes could be attriubted to the aging process and not the sex of the animal. It was concluded, however, that the strength of the insertion sites of ligaments on bones are responsive to the hormonal fluctuations that occur with aging.

Sports injuries at the 1985 Junior Olympics. An epidemiologic analysis.

The XIX Junior Olympic Games, hosted in Iowa City, Iowa, in August of 1985, involved 3,028 athletes who participated for 7 days in 13 different sports at 8 separate sites. Medical coverage for the Games was provided by the University of Iowa Sports Medicine Service. Staffing for the events involved approximately 75 physicians, 60 athletic trainers, and other health care personnel. A triage protocol was established prior to the Games whereby the athletic trainer would make first contact with the injured athlete and would evaluate and treat the injury based on standing orders. If in the judgment of the trainer, the athlete needed to be referred to a physician, one would be available, either on site or on call. During the Games, 1,113 medical encounters were recorded, 121 of those being deemed serious enough to withhold an athlete from competition pending further evaluation and treatment. The 121 significant injuries and illnesses involved 116 athletes (66% male, 34% female). Thirty-four percent of the significant injuries only required treatment by the trainer, while 46% were referred to an on site physician and 20% needed a specialty consultant. Seventeen percent of the significant injuries resulted in the athlete being medically disqualified for the remainder of the event. The most common injuries/illnesses were contusions (26%), sprains (21%), heat (17%), strains (9%), and other illnesses (12%). According to body region, 44% of the 121 injuries were to the lower extremity, 26% to the head, neck, and trunk, and 12% to the upper extremity.(ABSTRACT TRUNCATED AT 250 WORDS)

Head and neck injuries in college football: an eight-year analysis.

The present study documented head and neck injuries in a study group of 342 college football players at a single institution for a period of 8 years. All freshmen players were screened for evidence of: (1) past history of head and neck injuries, and (2) abnormalities of the cervical spine on physical examination and x-ray film. By recording all head injuries and those neck injuries with time loss, incidence rates and patterns of injury incurred in college competition were determined. A total of 175 head and neck injuries were sustained by 100 players over the 8 year period. Those players with abnormal findings on screening examination were twice as likely to have a head or neck injury at some point in their college careers as those players with a normal screening examination. The greater the degree of abnormality on freshman screening examination, the more severe the neck injury in college was likely to be. Twenty-nine percent of all players in the study group sustained a head or neck injury during their college careers. The probability of a subsequent head or neck injury escalated sharply following a single incident. The overall incidence of injury was found to have been dramatically reduced over the 8 years. Influential factors such as legislative rule changes, medical status of recruits, and general coaching philosophies are discussed with regard to injury reduction and prevention of head and neck injuries in college football.

Distribution and deposition of tritiated cortisol using phonophoresis.

Tritiated cortisol cream was coupled with light-microscopic autoradiography to histologically trace the distribution and deposition of a topical application of cortisol driven in by a therapeutic dose of ultrasound. Fifteen dogs were divided into four distinct groups: Group 1 consisted of two control dogs; Group 2, four dogs that received 10% radioactive cortisol and no ultrasound; Group 3, four dogs that received 5% radioactive cortisol plus ultrasound; and Group 4, five dogs that received 10% radioactive cortisol plus ultrasound. Although the cortisol applied to all dogs was limited to the epidermis, penetration occurred beyond the stratum corneum. The combination of 10% cortisol and ultrasound showed more penetration than 10% cortisol alone (p = .018), suggesting that ultrasound may be useful in the transdermal delivery of cortisol cream. The difference between the penetration of 5% and 10% cortisol when treated with ultrasound was only marginally significant (p = .062).

Who should manage continuous renal replacement in the intensive care setting? A nursing viewpoint.

Since its inception, continuous renal replacement therapy (CRRT) has been performed in critical care units with or without the involvement of nephrology nursing support (1,2). It is apparent that the issue of providing care to patients requiring this therapy is not so much a debate on the nursing control of CRRT, but a focused discussion on the nursing management and delivery of care to the patient receiving CRRT in the intensive care setting. Although the choice of a nursing care model for CRRT is dependent on many clinical and organisational factors, the use of one nursing specialty to deliver CRRT care can leave gaps in practice. The Joint or Collaborative Nephrology/critical care nursing model brings the highest level of nursing expertise to the bedside. The joint model tends to promote collaboration between two distinct nursing specialties, with opportunities for setting joint standards and promoting research. With this in mind, this discussion will examine some of the factors affecting structuring of nursing care, describe nursing models currently in use, compare the attributes of each, and conclude which model is preferred for the delivery of nursing care for CRRT.

Digital processing of microradiographic and fluorochromic images from bone biopsy.

Bone biopsy is an essential technique for the diagnosis of metabolic bone disease and the skeletal response to remedial therapeutic maneuvers. Among many approaches in analyzing the biopsy specimen, the imaging technique offers the greatest potential. The objective of this paper is to describe the methodologies developed for digital processing of microradiographic and fluorochromic images from bone biopsy. This procedure requires a microscope with conventional and ultraviolet illuminations, a vidicon camera with three color filters, a general purpose image processing system with three image memories, a digital computer with some specially tailored software programs. A good specimen preparation from bone biopsy is a prerequisite in assuring the success of the method. The methodology for processing the microradiographs is based on a completely developed procedure of black and white image processing. Production run is in progress. The methodology for processing the fluorochromic images is based on true color imaging, although the procedure is still preliminary, early results obtained so far are promising.