Transcriptomic response to differentiation induction.

BACKGROUND: Microarrays used for gene expression studies yield large amounts of data. The processing of such data typically leads to lists of differentially-regulated genes. A common terminal data analysis step is to map pathways of potentially interrelated genes. METHODS: We applied a transcriptomics analysis tool to elucidate the underlying pathways of leukocyte maturation at the genomic level in an established cellular model of leukemia by examining time-course data in two subclones of U-937 cells. Leukemias such as Acute Promyelocytic Leukemia (APL) are characterized by a block in the hematopoietic stem cell maturation program at a point when expansion of clones which should be destined to mature into terminally-differentiated effector cells get locked into endless proliferation with few cells reaching maturation. Treatment with retinoic acid, depending on the precise genomic abnormality, often releases the responsible promyelocytes from this blockade but clinically can yield adverse sequellae in terms of potentially lethal side effects, referred to as retinoic acid syndrome. RESULTS: Briefly, the list of genes for temporal patterns of expression was pasted into the ABCC GRID Promoter TFSite Comparison Page website tool and the outputs for each pattern were examined for possible coordinated regulation by shared regelems (regulatory elements). We found it informative to use this novel web tool for identifying, on a genomic scale, genes regulated by drug treatment. CONCLUSION: Improvement is needed in understanding the nature of the mutations responsible for controlling the maturation process and how these genes regulate downstream effects if there is to be better targeting of chemical interventions. Expanded implementation of the techniques and results reported here may better direct future efforts to improve treatment for diseases not restricted to APL.

Nicotine modulation of cytokine induction by LPS-stimulated human monocytes and coronary artery endothelial cells.

Nicotine, the major addictive component of tobacco, is an immunomodulator that impacts on many cells, including immune cells involved in inflammatory processes. Nicotine also induces oxidative damage to the vascular endothelium and accentuates lipid peroxidation, resulting in vascular cell dysfunction. Furthermore, vascular endothelial cells produce growth factors, such as cytokines and chemokines capable of stimulating and recruiting immune cells to atheromatous lesions. In addition, bacterial products including lipopolysaccharides (LPS), a major component of Gram negative bacterial cell walls, activate gene expression resulting in inflammatory cytokine production causing further damage to the vasculature. In the present study, the combined effects of nicotine and bacterial LPS on the expression of IL-6, IL-8, GRO-alpha and MCP-1 in cell lines of human coronary artery endothelial cells (HCAEC) and pulmonary monocytes (THP-1) were examined by quantitative real-time PCR and ELISA. Results showed that nicotine suppressed the LPS induced production of IL-6 and IL-8 in both cell lines. Since cytokines which alter homeostasis of both vascular endothelial and immune cells are critical for the atherogenic process, further studies are warranted to examine in detail the role of nicotine in terms of effects on inflammatory reactions, including those induced by bacterial infection.

Injury-induced NF-kappaB activation in the hippocampus: implications for neuronal survival.

Nuclear factor (NF)-kappaB p50 protein is involved in promoting survival in hippocampal neurons after trimethyltin (TMT)-injury. In the current study, hippocampal NF-kappaB activity was examined and quantitated from transgenic kappaB-lacZ reporter mice after chemical-induced injury. NF-kappaB activity was localized primarily to hippocampal neurons and significantly elevated over that in saline-treated mice between 4 and 21 days after TMT injection. Seven days after TMT injection, a timepoint of elevated NF-kappaB activity, gene expression in the hippocampus was studied by microarray analysis through comparison of expression profiles between treated nontransgenic and p50-null mice with their saline-injected controls. Seventeen genes increased in nontransgenic TMT-treated mice relative to saline-treated as well as showing no increase in p50-null mice, indicating a role for p50 in their regulation. One of these genes, the Na+, K+-ATPase-gamma subunit, was detected in brain for the first time. Several of the genes modulated by NF-kappaB are potentially related to neuroplasticity, providing additional evidence that this transcription factor is a neuroprotective signal in the hippocampus.

The performance of subcutaneously injected Fertinex when used as the sole gonadotropin for in vitro fertilization stimulation.

OBJECTIVE: To assess the performance of Fertinex (urofollitropin; Serono Laboratories, Norwell, MA) in stimulating the development of multiple follicles and initiating subsequent pregnancy in patients undergoing controlled ovarian hyperstimulation (COH) for IVF. DESIGN: Prospective observational study. SETTING: Private assisted reproductive technology (ART) center. PATIENT(S): Ninety-three women who underwent an IVF cycle of COH in which Fertinex was used as the sole gonadotropin. INTERVENTION(S): The COH protocol included Fertinex with leuprolide acetate down-regulation in the late luteal phase or the early follicular phase. Embryo transfer was performed after 3 days of culture. MAIN OUTCOME MEASURE(S): Stimulation parameters, embryologic data, and pregnancy rates (PRs). RESULT(S): Ninety-three patients underwent 97 cycles of COH, with a cancellation rate of 13.4%. The clinical PR was 42% per initiation, 49% per retrieval, and 51% per transfer. Shorter stimulation periods, lower estradiol levels either per follicle punctured or per oocyte retrieved, and slower embryo development were observed. Patients responded to Fertinex in three distinct stimulation patterns: escalating response with a peak estradiol level of < or =3,000 pg/mL, escalating response with a peak estradiol level of >3,000 pg/mL, or escalating response characterized by a drop in the estradiol level or a plateau before hCG administration. All three responses had similar PRs. CONCLUSION(S): Fertinex can be used successfully as the sole gonadotropin for COH in ART without compromising high PRs. Traditional estradiol response curves or cancellation criteria may not apply when Fertinex is used as the sole gonadotropin for COH in ART.

Oocyte donation program: initial experiences at Southeastern Fertility Institute.

From March 1994 to February 1996, 28 infertile couples participated in the oocyte donation program in 33 treatment cycles at the Southeastern Fertility Institute. Of the 31 cycles with embryo transfer, 15 cycles (48.4 percent) resulted in a clinical pregnancy with fetal heart beat by ultrasound. The spontaneous first trimester abortion rate was 3/15 (20 percent), multiple pregnancy rate 3/15 (20 percent), live birth rate 11/15 (73.3 percent) and delivery rate 12/15 (80 percent). It is recommended that oocyte donation procedure is a highly successful treatment option for women with ovarian failure or repeated unsuccessful trials of assisted reproductive technologies.

NR8383 alveolar macrophage toxic growth arrest by hydrogen peroxide is associated with induction of growth-arrest and DNA damage-inducible genes GADD45 and GADD153.

Breathing air exposes humans and other mammals to various toxic agents including oxidative contaminants associated with fine particles of less than 2.5 micron which may be deposited in the deep lung and have been implicated in the increased morbidity and mortality correlated with air pollution. Oxidative damage from inhaled particles may include damage to DNA, thereby adversely affecting the immunosurveillance provided by alveolar macrophages. Using the rat alveolar macrophage cell line NR8383, we demonstrated that cell proliferation was inhibited by exogenous hydrogen peroxide, an oxidant naturally produced in cellular respiration and phagocytosis. Mercaptosuccinate, a specific inhibitor of the antioxidant enzyme glutathione peroxidase, also inhibited cell growth. Genes known to be coordinatively regulated in response to growth arrest and DNA damage, GADD45 and GADD153, were induced compared to the housekeeping gene beta-ACTIN by equitoxic doses of hydrogen peroxide and mercaptosuccinate. Hydrogen peroxide treatment of cells in which glutathione peroxidase was inhibited by mercaptosuccinate resulted in even greater induction of both GADD genes. This approach using the NR8383 alveolar macrophage cell line provides a model for studying genotoxicity at the mechanistic level at which stress-responsive genes involved in growth arrest and DNA-damage response are modulated.

Public participation in radiological surveillance.

In 1989, Pacific Northwest National Laboratory developed a program, for the U.S. Department of Energy, to involve local citizens in environmental surveillance at the Hanford Site. The Community-Operated Environmental Surveillance Program was patterned after similar community-involvement efforts at the Nevada Test Site and the Three Mile Island nuclear facility. Its purpose is to increase the flow of information to the public, thereby enhancing the public's awareness and understanding of surveillance activities. The program consists of two components: radiological air monitoring at nine offsite locations and agricultural product sampling at selected locations near the site. At each air-monitoring station, two local school teachers collect air particulate samples and operate equipment to monitor ambient radiation levels. Atmospheric tritium samples (as water vapor) are also collected at some locations. Four of the air-monitoring stations include large, colorful informational displays for public viewing. These displays provide details on station equipment, sample types, and sampling purposes. Instruments in the displays also monitor, record, and show real-time ambient radiation readings (measured with a pressurized ionization chamber) and meteorological conditions. Agricultural products, grown primarily by middle-school-aged students, are obtained from areas downwind of the site. Following analysis of these samples, environmental surveillance staff visit the schools to discuss the results with the students and their teachers. The data collected by these air and agricultural sampling efforts are summarized with other routinely collected sitewide surveillance data and reported annually in the Hanford Site environmental report.

Ambient air sampling for tritium--determination of breakthrough volumes and collection efficiencies for silica gel adsorbent.

Ambient air samples for tritium (as HTO) can be collected using the solid adsorbent silica gel. The purpose of this study was to determine the maximum practical sampling volume and overall collection efficiency for water vapor collected on silica gel columns and to demonstrate the use of an impinger-based system to load water vapor onto silica gel columns. Breakthrough volumes (Vb) were measured and chromatographic efficiencies (expressed as the number of theoretical plates, N) were calculated for a 20 degrees C to 50 degrees C temperature range, with the relative humidity at approximately 30%. The tests yielded relative breakthrough volumes (air volume/adsorbent depth, m3 cm(-1)) of 0.36 for 20 degrees C, 0.20 for 30 degrees C, 0.15 for 40 degrees C, and 0.077 for 50 degrees C. For 18-cm columns, the average tritium tracer recoveries at 20 degrees C were 71% with no observed breakthrough for air volumes up to 5 m3, while at 40 degrees C mean tritium tracer recoveries dropped from 75% for volumes < or = 3.0 m3, to 0% for a volume of 5.0 m3. Frontal chromatographic profiles were measured for water vapor migrating through silica gel columns that were divided into 5 segments. The chromatographic efficiency of the silica gel columns was determined by graphical evaluation of the chromatography profiles. At a sampling rate of 0.25 L min(-1) and 30% relative humidity, the number of theoretical plates per adsorbent depth were 0.55 N cm(-1) at 20 degrees C, 0.68 N cm(-1) at 30 degrees C, 0.51 N cm(-1) at 40 degrees C, and 0.30 N cm(-1) at 50 degrees C. Chromatographic theory was used to estimate the overall collection efficiency of the silica gel columns as a function of the ratio of the sampling volume to breakthrough volume and the chromatographic efficiency. For a 9.5 m3 sample volume, 30% relative humidity, 0.25 L min(-1) sampling rate, and a 54-cm column, the overall collection efficiency was above 99.9% at 20 degrees C, above 95% at 30 degrees C, just below 80% at 40 degrees C, and <<80% at 50 degrees C.

A role for airborne particulates in high mercury levels of some cetaceans.

In a study of 25 Tursiops truncatus and two Globicephala macrorhynchus examined by necropsy, abundant HgSe was found in both the liver and in the respiratory system (lung and hilar lymph nodes). In the liver HgSe was consistently associated with the cell-breakdown pigment lipofuscin, whereas in lung and hilar lymph nodes it was consistently associated with particulates consisting of partially graphitic soot and silicates. This supports earlier suggestions that in the liver HgSe may be a storage end product of Hg metabolism, while adding the new suggestion that in the respiratory system HgSe may be inhaled, preformed in combustion emissions.

Transvaginal embryo transfer during the zygote intrafallopian tube transfer procedure.

OBJECTIVE: Our purpose was to evaluate a tactile non-ultrasonographically directed approach to transvaginal tubal cannulation and embryo transfer during the zygote intrafallopian tube transfer procedure. STUDY DESIGN: We studied the application of the tactile approach to transvaginal tubal cannulation and embryo transfer in 68 consecutive zygote intrafallopian tube transfer cycles. RESULTS: The tactile approach to transvaginal tubal cannulation and subsequent embryo transfer was successful in 93% of couples. Pregnancy occurred in 16 of the 63 (25%) women undergoing a successful transvaginal embryo transfer. CONCLUSION: The tactile approach to transvaginal tubal cannulation was found to be easily learned and provided a safe and convenient method of tubal embryo transfer. This low-cost approach, which has application to transfer of both embryos and gametes, deserves further study.

Office-based ankle arthroscopy.

Arthroscopy of the ankle has a definite role in the diagnosis and treatment of intra-articular pathologic conditions. Topics discussed in this article include advantages and disadvantages of arthroscopy, patient evaluation, indications for diagnostic arthroscopy, and techniques and considerations of office-based arthroscopy. Four case studies supplement the text.

Effect of aging on prospective and incidental memory.

Three experiments were conducted to test the effect of aging on prospective and incidental memory. In the first study, 24 undergraduates (M = 19.71 years) and 17 Elderhostel participants (M = 69.06 years) were used to test three hypotheses: (1) Younger subjects will be superior to older subjects in incidental memory. (2) If older and younger subjects have access to external memory aids, older subjects will display significantly better prospective memory. (3) If these subjects are denied external aids, the younger subjects will exhibit better prospective memory. Hypotheses 1 and 2 were confirmed, but 3 was not. The advantage older subjects display when permitted use of memory aids could derive from better recall of the future task or higher motivation to complete the task once it is recalled. In the second experiment, 51 undergraduates (M = 18.71 years) and 55 older subjects (M = 71.87 years) recruited from a continuing education program were used to test the hypothesis that the advantage older subjects displayed in aided prospective memory over younger ones derives from better recall of the task. The results obtained were not consistent with this interpretation. In the third experiment, 22 undergraduates (M = 18.50 years) and 20 Elderhostel participants (M = 70.25 years) tested the hypothesis that the advantage older subjects enjoy in prospective memory derives from greater motivation to complete the task. This hypothesis was confirmed.

Liver abnormalities associated with chronic mercury accumulation in stranded Atlantic bottlenose dolphins.

Eighteen stranded Atlantic bottlenose dolphins (Tursiops truncatus) examined postmortem were sampled for histologic study. All cases were examined for ferric ion and lipofuscin. Ages were determined from tooth growth layers. Electron microscopic (EM) examination and X-ray spectroscopy (EDAX) were performed. Chemical analysis for mercury was conducted on 12 of the animals by atomic absorption spectrophotometry. Nine animals were found to have excessive lipofuscin in both liver and kidney. Four of these nine animals also exhibited active liver disease (fat globules, central necrosis, lymphocytic infiltrates) whereas, of the animals without the excessive pigment, only one animal had an active liver lesion. EM and EDAX showed electron-dense amorphous material presumably within lysosomes to be Hg with no deposits on mitochondrial or nuclear membranes noted. Age relationship to portal pigment deposition was positive. Liver mercury concentrations ranged from 0.01 to 443 micrograms/g of wet weight with all animals having liver pigment yielding values of or above 61 micrograms/g, whereas all animals lacking pigment had values of or below 50 micrograms/g. The evidence suggests that the excessive pigment accumulation is related to toxic effects of Hg and presents as increased active liver disease.

Lymphangiomyomatosis in the Atlantic bottlenose dolphin (Tursiops truncatus).

The first case of lymphangiomyomatosis in a marine mammal is reported from a stranded male Atlantic bottlenose dolphin (Tursiops truncatus). This progressive proliferative disease involved the lungs and the mediastinal and probably mesenteric lymph nodes. An extraordinarily low level of testosterone may have been pathologically significant since all reported cases of this disease in humans occur in females.

Dual-image video endoscopy.

The use of a digital audio-video mixer is advocated when concurrent endoscopic procedures, such as laparoscopy and hysteroscopy, are performed. The device can also assist in postproduction work.

Use of an oxidized, regenerated cellulose absorbable adhesion barrier at laparoscopy.

The use of an oxidized, regenerated cellulose absorbable adhesion barrier has been shown to significantly reduce the incidence, extent and severity of postoperative gynecologic adhesions in a multicenter, randomized, prospective trial. Since an increasing proportion of gynecologic surgery is being performed laparoscopically, a prospective, multicenter, open-label study was initiated to evaluate the feasibility of using the adhesion barrier laparoscopically and to establish techniques for applying the barrier during those procedures. Eighty patients were studied: 42 underwent lysis of adhesions; 35, removal of endometriosis implants; and 14, tuboplasty/fimbrioplasty. An average of two pieces were placed per patient, with an average piece size of 3 x 3 cm and an average time required per piece of 2 minutes, 40 seconds. Seventy-five percent of the cases required less than three pieces each for application. Eighteen percent of the pieces were difficult to place, generally because of problems unfolding the material, excessive size or nonadherence. For similar reasons, replacement was necessary for 27 of 157 pieces placed (17.2%). The barrier can be applied with relative ease during operative laparoscopy. The fabric can be placed through either the suprapubic or umbilical sleeve, depending on the size of the piece of barrier required.

Collection of nonpolar organic compounds from ambient air using polyurethane foam-granular adsorbent sandwich cartridges.

Glass cartridges containing 6-10 g of Tenax-GC or 15 g of XAD-2 resin packed between two slices of polyurethane foam (PUF) were used in a General Metal Works PS-1 high-volume sampler to collect chlorobenzenes (CBs) containing three to six chlorine atoms, hexachlorocyclohexanes (HCHs), and two-ring aromatic hydrocarbons from 35-385 m3 of air. Laboratory experiments were run by vaporizing known quantities of analytes into a clean airstream for sampling by the PS-1 system at 20 degrees C. Collection efficiencies, determined from mass balance of the quantities introduced and recovered, ranged from 70 to 120% for individual compounds and averaged 93% overall. Penetration of analytes to backup adsorbent traps showed an inverse correlation to vapor pressure. The method was used to collect the above compounds from urban and rural air.

Absorbable pin: a new method of fixation for digital arthrodesis.

A new method of fixation of digital arthrodesis is being used at Atlanta Hospital. A total of 58 digits have been arthrodesed at the proximal interphalangeal joint using absorbable pin fixation. The technique of absorbable pin insertion, as well as the advantages and disadvantages, are discussed in this article. Infection, foreign body reaction, vascular compromise, or excessive swelling and edema were not evident in any of the 58 digits reviewed. The internal absorbable pin fixation is considered a viable alternative to previous forms of fixation for digital arthrodesis. The absorbable pin previously has been used for osteochondral fractures of the knee.

Rupture of the lateral head of the gastrocnemius muscle at the musculotendinous junction mimicking a compartment syndrome.

This case report documents an acute rupture of the lateral head of the gastrocnemius muscle at the myotendinous junction and a lateral dislocation of the tibialis anterior tendon with a clinical presentation consistent with a compartment syndrome, despite the intraoperative finding of a rupture of the posterior and lateral compartments. Extensive hematoma formation led to marked edema, paresthesias, muscle weakness, and severe pain in the involved leg. Surgical repair of the torn muscle and dislocated tendon and evacuation of the dissecting hematoma resulted in a well-functioning extremity. The authors emphasize the importance of prompt diagnosis of soft tissue injuries, which may place a patient at risk for a compartment syndrome. A compartment syndrome may lead to severe ischemia and irreversible tissue necrosis if intracompartmental hemorrhage of a torn muscle persists, and may require a surgical decompression. The clinical presentation, as well as adjunctive techniques in the diagnosis of a patient with a partial rupture of the gastrocnemius muscle, and a compartment syndrome, were presented.