[Relationship between body temperature, neuron-specific enolase, and clinical course in patients after out-of-hospital cardiac arrest]. / Zusammenhang zwischen Körpertemperatur, neurononspezifischer Enolase und klinischem Verlauf bei Patienten nach prähospitaler Reanimation.

BACKGROUND: According to ILCOR (International Liaison Committee on Resuscitation) recommendations (released in 2003), use of therapeutic hypothermia is recommended for unconscious adult patients who have survived a cardiac arrest regardless of the initial monitored cardiac rhythm. Thereby, the treatment goal is to achieve and maintain a body temperature of 32-34 °C for a period of 12-24 h. According to the October 2015 recommendations of the European Resuscitation Council (ERC), targeted temperature management (TTM) remains part of treatment, but, as an option, it is advised that the targeted body temperature be 36 °C rather than 32-34 °C. PATIENT POPULATION AND METHODS: For a non-randomized retrospective observational study, a total of 149 patients were treated with cardiopulmonary resuscitation (CPR) between May 1999 and September 2009. For the first 4 days after CPR, data associated with demography, resuscitation, therapy (temperature course, neuron-specific enolase [NSE]) and clinical-neurological development (Glasgow Outcome Scale [GOS]) were collected. In the study, patients receiving mild hyperthermia were compared with those who did not receive hypothermia. RESULTS: Of the 149 patients included, 90 were treated with mild hypothermia (as decided by the attending physician), while 59 received no hypothermia therapy. Assessment reveals that mild hypothermia positively influences clinical-neurological progression, but not survival. On day three and four, patients with an unfavorable neurological progression exhibited significantly increased serum levels of NSE (day 4: 108.7 ± 137.3 ng/ml versus 25.5 ± 15.4 ng/ml). Patients receiving hypothermia showed lower average NSE levels compared with persons not receiving hypothermia. Furthermore, during the first 4 days, their NSE values tended to increase slower (NSE value at day 4: 55.9 ± 64.9 ng/ml versus 129.9 ± 174.9 ng/ml). The best cut-off-value for an unfavorable neurological result was 74.2 ng/ml at day four (specificity 100%, sensitivity 48.6%). For the group of patients who received hypothermia, the best cut-off-value was 74.2 ng/ml at day four (specificity 100%, sensitivity 40.9%), and, for the comparison group, best cut-off-value was 25.5 ng/ml at day three (specificity 100%, sensitivity 88.2%). CONCLUSION: After out-of-hospital resuscitation, there is a trend for improved clinical-neurological progression with mild hypothermia but it does not influence the prognostic significance of serum NSE. After assessment of available data, it is not possible to recommend uniform cut-off values for patients who received mild therapeutic hypothermia and for those who did not receive hypothermia treatment.

Susceptibilidad antimicrobiana en muestras urogenitales de ureaplasma urealyticum, el principal agente en infección intramaniótica / Antimicrobial susceptibility in urogenital samples of ureaplasma urealyticum, the main agent in intramaniotic infection

RESUMEN OBJETIVO: Ureaplasma urealyticum es el agente más frecuentemente aislado en infección intraamniótica. Los macrólidos son los antimicrobianos de primera elección en embarazadas. Se ha descrito el aumento de resistencia, pudiendo limitar las opciones terapéuticas durante la gestación. El propósito del estudio es evaluar susceptibilidad antimicrobiana de Ureaplasma urealyticum aislado en mujeres en edad fértil, que se atienden en Clínica Alemana Temuco, Araucanía, Chile. METODO: Se estudian todas las muestras de orina y flujo vaginal para cultivo de U. urealyticum, de pacientes entre 18 y 40 años, recibidas en el Laboratorio de Microbiología Clínica Alemana Temuco, en período Abril 2013 a Enero 2015. Se procesan las muestras con kit Mycoplasma IST 2 de Biomerieux. En las que resultan positivas, se estudia susceptibilidad a macrólidos, tetraciclinas y quinolonas. RESULTADOS: 426 muestras de orina y flujo vaginal (390 pacientes). 197 pacientes resultaron positivas para U. urealyticum. (50,5%). La susceptibilidad fue 88,4% (174 pctes) a Eritromicina, 87,9% (173 pctes) a Claritromicina y 91,9% (181 pctes) a Azitromicina (NS). 15 de 197 pacientes (7,6%) fueron resistentes a los 3 macrólidos. La susceptibilidad a Quinolonas fue 55,3% a Ciprofloxacino, y 94% a Ofloxacino. El 100% resultó susceptible a Tetraciclinas. CONCLUSIONES: Cerca del 10% de U. urealyticum aislados en nuestra serie son resistentes a macrólidos, contribuyendo a la no erradicación de la infección en tratamientos empíricos. Dentro de ellos, azitromicina aparece con la mayor efectividad. El aumento de resistencia limitará opciones terapéuticas, con gran impacto perinatal en futuro. La vigilancia de susceptibilidad en cada hospital es fundamental para elección terapéutica.

ABSTRACT INTRODUCTION: Ureaplasma urealyticum is the most frequently isolated microorganism in intra-amniotic infection. The macrolides are the first choice antimicrobials for treat this infection in pregnancy. The increasing resistance has been described worldwide, seriously limiting therapeutic options in pregnancy. The aim of the study is to evaluate antimicrobial susceptibility of U. urealyticum aislated in fertile-age women in Clínica Alemana Temuco, Araucania region, Chile. METHOD: Urine and vaginal samples were analyzed for U. urealyticum, from every 18 to 40 years old patients, received at Microbiology Laboratory of Clínica Alemana Temuco, between April 2013 to January 2015. The samples are processed with Mycoplasma IST 2 kit of Biomerieux. If they became positives, susceptibility to macrolides, tetracyclines and quinolones was studied. RESULTS: 426 urine and vaginal samples were collected (390 patients). 197 patients were positive for U. urealyticum (50.5%). The susceptibility was 88.4% (174 pts) to Erythromicyn, 87.9% (173 pts) to Clarithromycin and 91.9% (181 pts) to Azithromycin (NS). Resistance to all macrolides was observed in 15 out of 197 patients (7.6%). The susceptibility to Quinolones was 55.3% to Ciprofloxacin, and 94% to Ofloxacin. The 100% was susceptible to Tetracyclines. DISCUSSION: Near to 10% of isolated Ureaplasma spp in our serie were resistant to some macrolide, being a factor for failing to eradicate the infection in empirical treatment. Azithromycin was the most effective. The increasing resistance will limit therapeutic options, with great perinatal impact in the future. Susceptibility surveillance in each hospital is very important for therapeutic options.

Current approaches to avoid the culling of day-old male chicks in the layer industry, with special reference to spectroscopic methods.

The negative correlation between fattening and laying performance prevents breeding improvement in both laying performance and meat yield. Therefore, specialized chicken lines have been bred in order to achieve either an efficient production of high-quality eggs or high growth rates. As a result, day-old male chicks are culled in the layer hatchery, which poses animal welfare and ethical problems. Breeding companies, scientific groups, and hatcheries are attempting to resolve this issue, with a common aim to find feasible alternatives for the routine killing of male layer chicks. Some approaches aim to influence the sex ratio, while others target at the economically feasible use of the male layer offspring, such as the fattening of "laying hen brothers" or crossbreedings of layers and broilers to create "dual-purpose chickens." Another approach is the sex determination prior to hatch. One of the prerequisites of in ovo sex determination is a practicable method that can be used in industry. The analysis needs to be rapid, cost-efficient, and highly precise; in addition, negative impacts on hatching rate, animal health, and/or performance parameters should be limited. Furthermore, sex determination should be performed before the sensory nervous system's response of the chick embryo to certain or potentially harmful stimuli is developed, which according to current knowledge is before the d 7 of incubation.

Identification of maternal characteristics associated with the use of epidural analgesia.

The present survey aims to identify predictors associated with the use of epidural analgesia (EA). Therefore, from October 2007 to June 2008, a survey was conducted in 193 pregnant women (mean age 31.7 years (SD 4.9); 64.8% primipara) attending a German general hospital with a specialisation in integrative medicine. Questionnaires, including Antonovsky's sense of coherence (SOC) were delivered antepartum. Delivery data were recorded within the hospital quality management programme. The adjusted odds ratio (OR) for EA use was significantly greater than one for women who had previously used EA (adjusted OR =4.1; CI: 1.03-16.31) and for the desire for a delivery without pain (adjusted OR =3.05; CI: 1.36-6.83). The likelihood of EA use decreased in multipara (adjusted OR =0.05; CI: 0.01-0.22). SOC was not found to be an independent predictor for EA use. However, women with high SOC more often preferred a delivery without EA (p for trend =0.037). In conclusion, first time labour, the desire for a delivery without pain and previous use of EA are independent predictors for the use of EA in labour. Further studies should clarify the predictive role of SOC in pregnancy.

Minimal doses of a sequence-optimized transgene mediate high-level and long-term EPO expression in vivo: challenging CpG-free gene design.

Advanced gene delivery techniques can be combined with rational gene design to further improve the efficiency of plasmid DNA (pDNA)-mediated transgene expression in vivo. Herein, we analyzed the influence of intragenic sequence modifications on transgene expression in vitro and in vivo using murine erythropoietin (mEPO) as a transgene model. A single electro-gene transfer of an RNA- and codon-optimized mEPOopt gene into skeletal muscle resulted in a 3- to 4-fold increase of mEPO production sustained for >1 year and triggered a significant increase in hematocrit and hemoglobin without causing adverse effects. mEPO expression and hematologic levels were significantly lower when using comparable amounts of the wild type (mEPOwt) gene and only marginal effects were induced by mEPOΔCpG lacking intragenic CpG dinucleotides, even at high pDNA amounts. Corresponding with these observations, in vitro analysis of transfected cells revealed a 2- to 3-fold increased (mEPOopt) and 50% decreased (mEPOΔCpG) erythropoietin expression compared with mEPOwt, respectively. RNA analyses demonstrated that the specific design of the transgene sequence influenced expression levels by modulating transcriptional activity and nuclear plus cytoplasmic RNA amounts rather than translation. In sum, whereas CpG depletion negatively interferes with efficient expression in postmitotic tissues, mEPOopt doses <0.5 µg were sufficient to trigger optimal long-term hematologic effects encouraging the use of sequence-optimized transgenes to further reduce effective pDNA amounts.

Dynamics of S100B release into serum and cerebrospinal fluid following acute brain injury.

High S100B serum levels are considered to reflect brain injury severity. However, the dynamics of S100B passage from the cerebral compartment into the blood remain unclear. We examined the temporal profile of S100B release into the cerebrospinal fluid (CSF) and blood in acute brain injury.In patients treated with ventricular drainage (subarachnoid hemorrhage, SAH, n = 23; traumatic brain injury, TBI, n = 19), we measured S100B levels in the serum and CSF. The Glasgow Coma Score (GCS) was assessed daily. Statistical analysis was performed by the Mann-Whitney rank sum test for group differences and by the Pearson correlation.In normal controls (n = 6), S100B levels in the serum (0.05 +/- 0.01 microg/L) comprised around 10% of the CSF concentration (0.66 +/- 0.08 microg/L). Following brain injury, S100B levels were significantly increased in the serum (p < 0.05 in SAH day 2-5, TBI day 1-8) and excessively increased in the CSF (p < 0.05 in SAH and TBI day 1-10). For the individual patient, there was no consistent correlation between S100B levels in serum or CSF and GCS. We therefore calculated the ratio of S100B serum/CSF. Following brain injury, the S100B passage from the CSF to the blood was significantly impaired. Further, higher ratios were correlated with better neurological function (p = 0.002).Because stimulated active S100B release may serve as a repair mechanism, a higher S100B serum/CSF ratio may contribute to neurological recovery.

Partonic pole matrix elements for fragmentation.

A model-independent analysis of collinear three-parton correlation functions for fragmentation is performed. By investigating their support properties it is shown, in particular, that the so-called partonic pole matrix elements vanish. This sheds new light on the understanding of transverse single spin asymmetries in various hard semi-inclusive reactions. Moreover, it gives additional strong evidence for the universality of transverse-momentum-dependent fragmentation functions.

Analysis of murine vascular function in vivo by optical coherence tomography in response to high-fat diet.

In this study, we demonstrate the application of optical coherence tomography (OCT) as a contactless imaging technique to analyze vasodynamics in small blood vessels in vivo. The transluminal OCT imaging of vessels avoids micro traumata in the endothelium and circumvents surgical intervention. It can be performed in the intact perfused vessel and provides a new method to measure vascular function and dynamics in vivo. The resolution of 10 mum and the velocity of image acquisition are adequate to detect differences in the inner diameter, the maximal velocity, or the time to half-maximal diameter change of small vessels. We applied this new technology to study the vascular dynamics in small vessels of 6- and 20-week-old C57BL/6 mice in vivo. In addition, we determined by this technique the impact of a high-fat diet for 14 weeks on vascular function in 20-week-old animals. The diameter of the saphenous artery was increased under resting conditions, after vasoconstriction and after vasodilatation in 20-week-old animals on normal chow and high-fat diet, compared to 6-week-old animals. High-fat diet caused a significantly impaired vasoconstriction in the saphenous artery. The maximal velocity of diameter changes of the saphenous artery was determined by time-resolved OCT imaging. A significant reduction of this parameter was found during vasoconstriction in 20-week-old mice on high-fat diet, compared to 6-week-old animals. In conclusion, transluminal optical coherence tomography imaging is a novel and useful technique to analyze the impaired vasodynamics of small arteries in response to high-fat diet in vivo.

[Biochemical bone resorption markers during the healing of osteoporotic fractures]. / Biochemische Knochenresorptionsmarker in der Heilungsphase osteoporotischer Frakturen.

BACKGROUND: This study investigated the progression and clinical relevance of biochemical resorption marker values during fracture healing in osteoporosis. PATIENTS AND METHODS: In 44 patients with distal radius fractures and 29 patients without fractures, the blood and urine concentrations of pyridinoline (PYD), deoxypyridinoline (DPD), N-telopeptides (NTx), and bone sialoprotein (BSP) were recorded on the day of trauma as well as during further progression. All postmenopausal patients underwent bone density measurement. Accordingly, patients were divided into premenopausal, postmenopausal osteoporotic, and postmenopausal nonosteoporotic groups. RESULTS: Between the groups, PYD, DPD, and NTx showed significant differences in their initial values. However, their further relative progression was primarily affected by the chosen therapy. CONCLUSION: Bone resorption markers can diagnostically point to osteoporosis and are significant parameters in fracture healing.

Posteromedial approach to proximal tibia for corticotomy in callus distractions.

Corticotomy of the tibia using Ilizarov's anterolateral approach is used routinely for callus distraction. This method is associated with impaired callus formation and delayed healing because of marginal soft tissue covering and blood supply to the proximal tibia. We presumed a newly designed posteromedial approach would result in less callus defects and improved healing. In this prospective, randomized study, 31 patients had callus distraction using an anterolateral approach or the newly designed posteromedial approach. Callus formation was assessed radiographically and histologically. Callus defects were classified using serial radiographs. Biopsy specimens were taken from high-grade defect (Grades 3-4) zones to examine the osteogenic potential. Radiographic evaluation showed 13 callus defects; 12 occurred after the anterolateral approach and only one occurred after the posteromedial method. Although low-grade defects (Grades 1-2) healed spontaneously, biopsy specimens taken from Grades 3-4 defects revealed no osteogenic potential and requiring operative revision. Because of low soft tissue covering and impaired blood supply to the anterior tibia during surgical exposure for corticotomy, less callus formation occurred after the anterolateral approach compared with the posteromedial approach. We recommend the less invasive posteromedial approach to reduce callus defects and impaired healing in callus distraction of the proximal tibia.

[Frequency and severity of callus defects. Dorsomedial vs ventrolateral approach for corticotomy in performing callus distraction of the tibia]. / Häufigkeit und Schweregrad von Kallusdefekten. Dorsomedialer vs. ventrolateraler Zugangsweg für die Kortikotomie bei der Tibiakallusdistraktion.

BACKGROUND: The purpose of this study was to compare a dorsomedial to the ventrolateral approach for corticotomy in performing callus distraction of the proximal metaphyseal tibia. PATIENTS AND METHODS: A total of 31 callus distractions were performed in 28 humans. The ventrolateral approach was used for 18 and the dorsomedial approach for 13 corticotomies. A scale of four severity grades was used to classify callus defect zones based on their extent as evidenced on serial X-rays. Biopsies were taken from higher-grade defects (grades 3-4). RESULTS: A total of 13 radiological evaluations revealed 12 defects using the ventrolateral approach. Seven defects (grades 1-2) healed spontaneously, whereas six defects (grades 3-4) required operative intervention as histological tissue examination showed no osteogenic potential. CONCLUSION: To prevent callus defects of the proximal tibia in the future and to ensure maximal osteogenic potential in the distraction zone, a minimally invasive dorsomedial approach appears to achieve favorable results.

[Micro-CT analysis of cancellous bone fragments from the distal radius fracture zone in osteoporosis]. / Mikro-CT Analyse spongiöser Fragmente aus der distalen Radiusfrakturzone bei Osteoporose.

The incidence and prevalence of osteoporosis must be considered to continue to increase significantly due to the expected demographic development and environmental changes. In the diagnosis and staging of osteoporosis the three-dimensional bone structure should be as important as the bone mass or the mineral content of the bone. In this study, microfragments were taken from distal radius fracture zones and investigated in Micro-CT scans. Patients in which osteodensitometry of the lumbal spine had revealed osteoporosis in were found to have significantly reduced bone mass, bone density and trabecular thickness. Trabecular fractures which were found in non-osteoporotic patients even in robust trabeculae were detected by the two-dimensional analysis in thin locations and arborizations. Despite some trabeculae turned out to be very small the differences in the histomorphometry and the quality of trabecular fractures in osteoporotic as well as non-osteoporotic patients could be visualized very good in the Micro-CT analysis.

Signaling lymphocytic activation molecule (SLAM) is differentially expressed in human Th1 and Th2 cells.

We have used a real-time quantitative RT-PCR technique (TaqMan, PE Biosystems) to identify genes that are differentially expressed by human polarised CD4(+) T cell subsets (Th1 or Th2). The goal was to test the feasibility of the detection method in profiling the expression of a set of marker genes important for Th1 and Th2 differentiation. We demonstrate that in polarised human Th1 cells signaling lymphocytic activation molecule (SLAM), a member of the immunoglobulin superfamily, is expressed at 7-25-fold higher levels than in Th2 cells. Along with SLAM, expression of the IL-12 receptor chain beta 2 (IL-12R beta 2) and the IFN-gamma receptor chain beta (IFN-gamma R beta) proved to be useful molecular markers indicating the state of T cell polarisation, as previously reported. Treatment with IL-12 increased SLAM mRNA expression in T cells by 3-4-fold, whereas a number of other cytokines including PDGF-BB, IFN-alpha A, IFN-alpha A/D, IFN-beta, IFN-gamma or IL-9 had no effect. Stimulating T cells by co-ligating CD3 and CD28 increased SLAM protein surface expression in both Th1 and Th2 cells. In conclusion, real-time RT-PCR detection was found to be an accurate, sensitive and highly reproducible method for fast profiling of mRNA expression in Th1 and Th2 cell subsets.

Initial uptake kinetics in human skin exposed to dilute aqueous trichloroethylene in vitro.

In vitro uptake of 14C-labeled trichloroethylene (TCE) from dilute (approximately 5-ppb) aqueous solutions into human surgical skin was measured using accelerator mass spectrometry (AMS). We analyzed 105 breast-tissue samples obtained from three subjects, representing 27 separate exposure experiments conducted at approximately 20 degrees C for 0, 1, 5, 15, 30, or 60 min. The AMS data obtained positively correlate with (p approximately 0) and vary significantly nonlinearly with (p = 0.0094) exposure duration. These data are inconsistent (p approximately 0) with predictions made for TCE by a proposed U.S. Environmental Protection Agency (USEPA) dermal-exposure model, even when uncertainties in its recommended parameter values for TCE are considered, but are consistent (p = 0.17) with a 1-compartment model for exposed skin-surface tissue governed in vitro by a maximum effective permeability of K*p = 0.28 cm h-1 (+/- 7.0%) and a first-order rate constant of k1 = 1.2 h-1 (+/- 16%). The apparent compartment depth is estimated to be approximately 40-100 microns, i.e., to comprise much or all of the epidermis. In contrast, the USEPA model implies only negligible TCE penetration beyond SC during a 1-h exposure. The K*p estimate based on the 1-compartment model fit is consistent with estimates for TCE based on in vivo studies, which supports the hypothesis that the USEPA model underpredicts short-term dermal uptake of TCE from water. It is shown that for humans, this fit also implies that normalized total uptake of TCE from water by short-term dermal contact in vivo is predicted to be fK*p, where f is approximately 80% for longer normothermic exposures and approximately 95% during a brief hot shower or bath. This study illustrates the power of AMS to facilitate analyses of contaminant biodistribution and uptake kinetics at very low environmental concentrations.

Close proximity of c-erbA2 and c-erbA-beta genes on the short arm of chromosome 3.

The common loss region on the short arm of chromosome 3 (3p) in human breast tumors harbors two members of the c-erbA receptor gene family, c-erbA2 and c-erbA-beta, both of which recognize a BamHI polymorphism in human genomic DNA. Analysis of lymphocyte DNAs from 50 normal individuals and lymphocyte DNAs from 50 normal individuals and lymphocyte and tumor DNAs from 116 breast cancer patients revealed identical genotypes (a/a, b/b or a/b) for both probes. Furthermore, deletion of the same allele (a/- or -/b) of c-erbA2 and c-erbA-beta was detected in 25% of the 66 breast tumors from patients with constitutionally heterozygous genotypes for both genes. No sequence homology was detected between the c-erbA2 and c-erbA-beta genes, suggesting a physical linkage between these two genes. Digestion of the genomic DNA with combinations of restriction enzymes and hybridization with c-erbA2, which is a genomic fragment, and c-erbA-beta, which is a cDNA clone, provide evidence that c-erbA2 and a region of the c-erbA-beta gene are physically contiguous on the short arm of chromosome 3 and are separated by no more than 1.8 kb of DNA sequences.

An antisense oligodeoxynucleotide that depletes RI alpha subunit of cyclic AMP-dependent protein kinase induces growth inhibition in human cancer cells.

Enhanced expression of the RI alpha subunit of cyclic AMP-dependent protein kinase type I has been correlated with cancer cell growth. We provide evidence that RI alpha is a growth-inducing protein that may be essential for neoplastic cell growth. Human colon, breast, and gastric carcinoma and neuroblastoma cell lines exposed to a 21-mer human RI alpha antisense phosphorothioate oligodeoxynucleotide (S-oligodeoxynucleotide) exhibited growth inhibition with no sign of cytotoxicity. Mismatched sequence (random) S-oligodeoxynucleotides of the same length exhibited no effect. The growth inhibitory effect of RI alpha antisense oligomer correlated with a decrease in the RI alpha mRNA and protein levels and with an increase in RII beta (the regulatory subunit of protein kinase type II) expression. The growth inhibition was abolished, however, when cells were exposed simultaneously to both RI alpha and RII beta antisense S-oligodeoxynucleotides. The RII beta antisense S-oligodeoxynucleotide alone, exhibiting suppression of RII beta along with enhancement of RI alpha expression, led to slight stimulation of cell growth. These results demonstrate that two isoforms of cyclic AMP receptor proteins, RI alpha and RII beta, are reciprocally related in the growth control of cancer cells and that the RI alpha antisense oligodeoxynucleotide, which efficiently depletes the growth stimulatory RI alpha, is a powerful biological tool toward suppression of malignancy.

IRAOS: an instrument for the assessment of onset and early course of schizophrenia.

Since Kraepelin's first description of dementia praecox in 1889 many data and theories have been published on the onset and course of schizophrenia. Until now studies on these topics had to rely on first admission data and on the subsequent course of the disease. However, first hospitalisation is preceded by a wide variety of patterns and duration of the early course. Items taken from the pre-admission phase of the disease are often incorrectly used as premorbid characteristics, understandably preceding the subsequent course and outcome of schizophrenia with high predictive power. In relation to our interest to study the beginning of schizophrenia, systematically, paying special attention to the age and gender distribution of true onset and the symptomatology and pattern of the early and later course, we developed an 'Interview for the Retrospective Assessment of the Onset of Schizophrenia (IRAOS)'. It allows an objective, reliable, and valid assessment of the symptoms, psychological impairments, demographic and social characteristics as well as the referring points in time of the early course of psychosis. The instrument is administered as a semi-structured interview with both the patient and a key informant. The obtained information is extended by a systematic examination of the clinician's case notes. Some results derived from an ongoing study on age and gender differences in onset and patterns of early course are added to demonstrate the use of the instrument.

A cDNA encoding a muscle-type tropomyosin cloned from a human epithelial cell line: identity with human fibroblast tropomyosin TM1.

Tropomyosins (TM) expressed by human epithelial cells have only recently been characterized, and no sequence data for them has appeared. We cloned a cDNA encoding a high molecular weight, muscle-type TM from a LS174T human colon carcinoma epithelial cell cDNA library. On sequence analysis this cDNA (TMe1) was virtually identical to the previously reported sequence for human fibroblast TM1 encoded by the hTM beta gene. Expression of TM1/TMe1 mRNA and protein are low in epithelial cells compared with fibroblasts. The results indicate that cells of different developmental lineages (entodermal and mesodermal) can produce identical TM beta gene splice products while regulating expression of those transcripts in a lineage-specific way.

Cooperative effect of 8-Cl-cAMP and rhGM-CSF on the differentiation of HL-60 human leukemia cells.

In HL-60 leukemia cells the site-selective cAMP analog, 8-Cl-cAMP, at a dose of 5 microM produced growth inhibition with no signs of toxicity, whereas granulocyte-macrophage colony stimulating factor (GM-CSF) exerted an early transient increase of cell proliferation which was followed by differentiation toward monocytes. 8-Cl-cAMP in combination with GM-CSF blocked the growth stimulation due to GM-CSF and demonstrated a synergistic effect on the differentiation of HL-60 cells. The early proliferative effect of GM-CSF was correlated with an increased expression of type I regulatory subunit of cAMP-dependent protein kinase (RI alpha). Treatment with an RI alpha antisense oligodeoxynucleotide suppressed the GM-CSF-inducible cell proliferation and differentiation. Conversely, an RII beta antisense oligodeoxynucleotide, which suppresses the RII beta and causes a compensatory increase in RI alpha level, greatly enhanced the early proliferative input and the differentiation induced by GM-CSF. These results provide an insight into the mechanism of action of GM-CSF and the rationale for a combination differentiation therapy with 8-Cl-cAMP and GM-CSF.