Effects of different durations of sustained inflation during cardiopulmonary resuscitation on return of spontaneous circulation and hemodynamic recovery in severely asphyxiated piglets.

OBJECTIVE: We previously demonstrated that sustained inflation (SI) during chest compression (CC) significantly reduces time to return of spontaneous circulation (ROSC) when compared to 3:1 compression:ventilation (C:V) ratio during neonatal resuscitation. However, the optimal length of SI during CC to improve ROSC and hemodynamic recovery in severely asphyxiated piglets is unknown. AIM: To examine if different lengths of SI will improve ROSC and hemodynamic recovery in severely asphyxiated piglets. INTERVENTION AND MEASUREMENTS: Thirty newborn piglets (1-3 days) were anesthetized, intubated, instrumented and exposed to 30-min normocapnic hypoxia followed by asphyxia. Piglets were randomized into four groups: 3:1 C:V (n = 8), CC with an SI duration of either 20 s (CC+SI 20) (n = 8) or 60 s (CC+SI 60) (n = 8), and a sham group (n = 6). Cardiac function, carotid blood flow, cerebral and renal oxygenation as well as respiratory parameters were continuously recorded throughout the experiment. MAIN RESULTS: When compared with 3:1 group, both CC+SI 20 and CC+SI 60 groups had significantly shorter ROSC time (p = 0.002). All three intervention groups had similar hemodynamic recovery by the end of 4 h observation period. There was no difference in lung injury markers among all experimental groups. However, when compared to the sham group, the concentrations of IL-6 (thalamus) and IL-6 + IL-8 (frontoparietal cortex) of the 3:1 C:V group were significantly higher, respectively. CONCLUSIONS: Even though relatively less animals achieved ROSC, CC during SI significantly improved ROSC time compared to 3:1 C:V in asphyxiated newborn piglets. However, there was no difference in ROSC characteristics and hemodynamic recovery between two CC+SI groups.

Tidal volume delivery during continuous chest compressions and sustained inflation.

OBJECTIVE: To determine the distending pressure needed to achieve sufficient tidal volume (VT) delivery during continuous chest compressions (CC) superimposed by sustained inflation (SI) (CC+SI). DESIGN: Randomised animal/manikin trial. SETTING: University laboratory. SUBJECTS: Cadaver piglets/manikin. INTERVENTIONS: SI distending pressures of 5, 10, 15, 20, 25 and 30 cm H2O were delivered in random order during CC+SI for 2 min each. MAIN OUTCOME MEASURES: VT, gas flow and airway pressure. Spearman's r for distending pressure and VT. RESULTS: Distending pressure and VT correlated in cadaver piglets (r=0.83, p<0.001), manikin (r=0.98, p<0.001) and combined data (r=0.49, p<0.001). VT was delivered during chest recoil during CC in both models. In cadaver piglets, a distending pressure ∼25 cm H2O was needed to achieve an adequate VT. CONCLUSIONS: Chest recoil generates VT depending on an adequate distending pressure. This has previously been demonstrated in adult animals. A pressure of ∼25 cm H2O is needed to achieve an adequate VT delivery.

Saturated fatty acid induces cancer stem cell-like properties in human hepatoma cells.

Hepatic steatosis has been reported to be a risk factor for the development of liver cancer. The underlying mechanism for carcinogenesis remains to be elucidated. It has been postulated that cancer stem cells (CSCs) within tumor tissues are a subset of cells with stem cell properties of self-renewal and undifferentiation. The purpose of this study was to investigate the effects of a saturated fatty acid, palmitate (PA), on CSC-like properties of human hepatoma HepG2 cells. We investigated the effects of PA on HepG2 cells and primary rat hepatocytes (PRH) by exposing them to PA to induce lipid accumulation. Significant fat accumulation was observed by Oil Red O staining in cells exposed to PA, and it was accompanied by significant increase in NFκB (p65) nuclear translocation in HepG2 cells. Notably, PA significantly enhanced the sphere forming ability of HepG2 cells, but not PRH. Furthermore, PA significantly increased stemness gene expressions of Sox2 and Oct4, and sonic hedgehog (Shh) production. Notably, NFκB inhibitors, N-Acetyl-L-cysteine and pyrollidine dithiocarbamate, and a NOX inhibitor, diphenyleneiodonium, significantly attenuated PA-induced sphere forming ability of HepG2 cells. Our results suggest that lipid accumulation may not only induce pro-inflammatory responses in hepatocytes but may also activate CSC-like properties of hepatoma cells through NFκB activation.

Complementary effects of auricular acupressure in relieving constipation symptoms and promoting disease-specific health-related quality of life: A randomized placebo-controlled trial.

OBJECTIVES: Constipation has been identified as a worldwide health problem among elderly people. Currently, it is not effectively relieved by the use of laxatives and lifestyle modification. Previous studies reported promising results in managing constipation with auricular acupressure (AA), although its effectiveness was not affirmed. This study is to evaluate the complementary effects of AA in relieving constipation symptoms and in promoting disease-specific health-related quality of life (HRQOL) among elderly residential care home (RCH) residents in Hong Kong. DESIGN: Randomized placebo-controlled trial. SETTING: Elderly RCH. INTERVENTION: Ninety-nine participants were randomly assigned to either experimental group (AA using auricular plasters with magnetic pellets), placebo-controlled group (AA using auricular plasters with Semen Vaccariae), or usual care group (AA using auricular plasters only). AA was applied onto seven auricular acupoints for 10 days. MAIN OUTCOME MEASURES: Constipation symptoms and disease-specific HRQOL were measured before AA, at the completion of AA (D10), and at the 10th-day follow-up time (D20). RESULTS: Significant group×time interaction effect was found in the change of satisfaction subscale between the experimental group and placebo-controlled group at D10 (p=0.016) and D20 (p=0.016) relative to the baselines. For both constipation symptoms and disease-specific HRQOL, the experimental group demonstrated the greatest improvement after receiving AA at both D10 and D20 compared with the other two groups. CONCLUSION: The current findings indicated positive clinical value of AA with magnetic pellets in managing constipation in elderly RCH residents. AA was also found to be a safe and acceptable intervention.

Effect of cardiac output changes on exhaled carbon dioxide in newborn piglets.

INTRODUCTION: International neonatal resuscitation guidelines recommend that correct tube placement should be confirmed by clinical assessment and exhaled CO2 detection. Absence of exhaled CO2 after intubation suggests oesophageal intubation, non-aerated lungs, low tidal volume delivery, or low cardiac output. The relationship between changes in cardiac output and exhaled CO2 in neonates is unknown. The aim of the study was to determine if changes in cardiac output affect exhaled carbon dioxide in a porcine model of neonatal resuscitation. METHOD: Term piglets (n=5) aged 3-4 days were anesthetised, intubated, instrumented and exposed to normocapnic hypoxia. Exhaled CO2 was continuously measured using a flow sensor (Respironics NM3(®)). Pulmonary artery blood flow, a surrogate for cardiac output was measured using an ultrasonic flow probe (Transonic(®)). A semi-quantitative CO2-detector (Pedi-Cap(®)) was placed between the tracheal tube and flow sensor to assess colour change at changing levels of cardiac output. RESULTS: Median (IQR) pulmonary artery blood flow significantly decreased from 177 (147-177)mL/kg/min at baseline to 4 (3-26)mL/kg/min during hypoxia (p=0.02). Exhaled CO2 remained similar throughout the experiment, 47 (41-47)mmHg at baseline vs. 40 (38-41)mmHg at the end of the hypoxia (p=1.00). Additionally, at each time point, colour change at the Pedi-Cap(®) was observed. CONCLUSION: A significant decrease in cardiac output was not associated with changes in exhaled CO2 or failure to achieve a Pedi-Cap(®) colour change.

Characteristics of patients with Clostridium difficile infection in Taiwan.

The medical records of 84 patients with stool cultures positive for Clostridium difficile during the period August 2007 to June 2009 were retrospectively reviewed. A case of confirmed (toxigenic)C. difficile infection (CDI) was defined by the presence of symptoms (fever, diarrhoea, abdominal discomfort or distension, ileus) and the presence of toxigenic C. difficile. Patients with compatible clinical symptoms and stool cultures positive for non-toxigenic C. difficile isolates were defined as probable (non-toxigenic) CDI cases. Of these 84 patients, 50 (59.5%) were diagnosed as confirmed CDI and 34 (40.5%) as probable CDI. Thirteen (15.5%) of the 84 patients died during their hospital stay. Usage of proton pump inhibitors was a significant independent risk factor for CDI (OR 3.21, P=0.014). Of the 50 isolates associated with confirmed CDI, seven (8.3%) carried binary toxin genes (cdtAB), and six (7.1%) had a deletion in the tcdC gene. The mortality rate in confirmed CDI patients with isolates exhibiting deletion in the tcdC gene (2/6, 33.3%), those with isolates harbouring binary toxin genes (2/7, 28.6%), and those with isolates containing mutations in gyrA (2/7, 28.6%) and gyrB (1/2, 50%) was higher than the overall mortality rate (10/50, 20%) in patients with confirmed CDI.

Dosimetric advantages of generalised equivalent uniform dose-based optimisation on dose-volume objectives in intensity-modulated radiotherapy planning for bilateral breast cancer.

OBJECTIVE: We compared and evaluated the differences between two models for treating bilateral breast cancer (BBC): (i) dose-volume-based intensity-modulated radiation treatment (DV plan), and (ii) dose-volume-based intensity-modulated radiotherapy with generalised equivalent uniform dose-based optimisation (DV-gEUD plan). METHODS: The quality and performance of the DV plan and DV-gEUD plan using the Pinnacle(3) system (Philips, Fitchburg, WI) were evaluated and compared in 10 patients with stage T2-T4 BBC. The plans were delivered on a Varian 21EX linear accelerator (Varian Medical Systems, Milpitas, CA) equipped with a Millennium 120 leaf multileaf collimator (Varian Medical Systems). The parameters analysed included the conformity index, homogeneity index, tumour control probability of the planning target volume (PTV), the volumes V(20 Gy) and V(30 Gy) of the organs at risk (OAR, including the heart and lungs), mean dose and the normal tissue complication probability. RESULTS: Both plans met the requirements for the coverage of PTV with similar conformity and homogeneity indices. However, the DV-gEUD plan had the advantage of dose sparing for OAR: the mean doses of the heart and lungs, lung V(20) (Gy), and heart V(30) (Gy) in the DV-gEUD plan were lower than those in the DV plan (p<0.05). CONCLUSIONS: A better result can be obtained by starting with a DV-generated plan and then improving it by adding gEUD-based improvements to reduce the number of iterations and to improve the optimum dose distribution. Advances to knowledge The DV-gEUD plan provided superior dosimetric results for treating BBC in terms of PTV coverage and OAR sparing than the DV plan, without sacrificing the homogeneity of dose distribution in the PTV.

Differential hemodynamic effects of levosimendan in a porcine model of neonatal hypoxia-reoxygenation.

BACKGROUND: Neonatal asphyxia can be complicated by myocardial dysfunction with secondary alterations in pulmonary and regional hemodynamics. Levosimendan is a calcium-sensitizing inotrope that may support cardiac output, but little is known regarding its differential hemodynamic effects in asphyxiated neonates. METHODS: Mixed breed piglets (1-4 days old, weight 1.6-2.3 kg) were acutely instrumented. Normocapnic alveolar hypoxia (10-15% oxygen) was induced for 2 h, followed by reoxygenation with 100% (1 h) and then 21% oxygen (3 h). At 2 h of reoxygenation, after volume loading (Ringer's lactate 10 ml/kg), either levosimendan (0.1 or 0.2 µg/kg/min) or D(5)W (placebo) was infused for 2 h in a blinded, block-randomized fashion (n = 7-8/group). The systemic, pulmonary and regional (carotid, superior mesenteric and renal) hemodynamics were compared. RESULTS: At 0.1 and 0.2 µg/kg/min, levosimendan significantly increased cardiac output (121 and 123% of pretreatment, respectively) and heart rate, and decreased systemic vascular resistance without causing hypotension. Pulmonary arterial pressure and estimated pulmonary vascular resistance were significantly increased from pretreatment baseline in 0.1 but not 0.2 µg/kg/min levosimendan. Levosimendan infusion had no effects on regional hemodynamics. Myocardial efficiency but not oxygen consumption increased with 0.1 µg/kg/min levosimendan without significant effects on plasma troponin and myocardial lactate levels. CONCLUSIONS: In newborn piglets following hypoxia-reoxygenation injury, levosimendan improves cardiac output but has no marked effects in carotid, superior mesenteric and renal perfusion. It appears that various doses of levosimendan increase the cardiac output through different mechanisms. Further investigations are needed to examine the effectiveness of levosimendan as a cardiovascular supportive therapy either alone or in conjunction with other inotropes in asphyxiated neonates.

Dual arc volumetric-modulated arc radiotherapy (VMAT) of nasopharyngeal carcinomas: a simultaneous integrated boost treatment plan comparison with intensity-modulated radiotherapies and single arc VMAT.

AIMS: To compare the performance of volumetric-modulated arc radiotherapy (VMAT) by dual arc with fixed beam intensity-modulated radiotherapies (IMRTs) and single arc VMAT on nasopharyngeal carcinomas (NPC). MATERIALS AND METHODS: Twenty NPC cases were re-planned using the planning system of the Pinnacle(3®)SmartArc (SA) module to compare the performance of the following four techniques: seven-field (7F) and 18-field (18F) fixed beam IMRT, and single (SA(1)) and dual arc VMAT (SA(2)). The plan was delivered on an Elekta Synergy™ Linac equipped with an 80-leaf 1cm multileaf collimator. Three dose levels of planning target volumes (PTVs) with 70/59.4/54.0Gy in 33 fractions were prescribed and delivered as a simultaneous integrated boost. The conformity index and homogeneity index of the PTVs, the comprehensive quality index (CQI), the normal tissue complication probability for the organs at risk (OARs), and the planning time, delivery efficiency and accuracy were analysed. RESULTS: A significantly inferior conformity index at the three dose levels of PTV and homogeneity index of PTV(70) were observed in SA(1) compared with the other techniques. Comparable conformity index and homogeneity index of the PTV were observed among 7F/18F IMRT and SA(2). Based on the CQI of the 11 OARs, the most efficient dose reduction was observed in 18F IMRT followed in order by SA(2), 7F IMRT and SA(1). The planning time was on average 13.2/24.9/40.1/42.8min for 7F/18F IMRT/SA(1)/SA(2), respectively. With regards to the delivery efficiency compared with 7F IMRT, a 51 and 41% reduction in delivery time was achieved by SA(1) and SA(2), respectively. All techniques presented a high quality assurance pass rate (>98%) of the Γ(3mm,3%) criterion. CONCLUSION: In NPC cases, SA(2) gave superior results in terms of PTV coverage and OAR sparing compared with SA(1) and approached the performance achieved by 18F IMRT, but without sacrificing the delivery efficiency.

Sacrococcygeal teratoma in adults: case report and literature review.

Sacrococcygeal teratoma is one of the most common tumours in infants but rare in adults. We present a case of sacrococcygeal teratoma in a female adult. The clinical presentation, radiological and histological findings, management, and outcome are described.

Elimination of keratin artifact bands from western blots by using low concentrations of reducing agents.

We encountered beta-mercaptoethanol-dependent artifact signals in western blot analyses using polyclonal antisera. Replacing beta-mercaptoethanol with dithiothreitol in the loading buffer did not eliminate the artifact signals. However, lowering the concentration of either dithiothreitol or beta-mercaptoethanol eliminated the background problems and allowed specific detection of the target protein. These results are consistent with the background signal being caused by anti-keratin antibodies in the antisera and keratin contamination of reagents. This study highlights the importance of testing a range of reducing agent concentrations when trying to eliminate artifact bands from western blots. However, this method may not be applicable when target proteins have disulfide bridges.

Evaluation of two short tandem repeat multiplex systems for post-haematopoietic stem cell transplantation chimerism analysis.

INTRODUCTION: The follow-up of chimerism status after allogeneic haematopoietic stem cell transplantation (HSCT) is essential to predict successful engraftment to assess the development of graft-versus-host disease, graft rejection and disease relapse. Analysis of short tandem repeats (STR) via polymerase chain reaction is frequently used for chimerism determination. However, most commercially-available kits have been designed for forensic purposes and may not be optimal for chimerism analysis. The present study aims to identify suitable STR markers for patient-donor pairs of predominantly Malay and Chinese ethnicity using two commercially-available forensic kits. METHODS: We analysed six STR loci, namely, CSF1PO, TPOX, TH01 (using the CTT multiplex system) and F13A01, FESFPS and vWA (using the FFv multiplex system) in 33 human leukocyte antigen-matched Malaysian patient-donor pairs to determine the suitability of these two multiplex systems for chimerism analysis in our local population. RESULTS: Informativity (different alleles in donor and recipient) of each individual locus was TH01 73 percent, vWA 73 percent, F13A01 52 percent, CSF1PO 61 percent, FESFPS 39 percent and TPOX 45 percent. When combined, the six STR loci were able to give chimerism results in 31 out of 33 (94 percent) cases. CONCLUSION: We found that TH01 and vWA are informative STR targets for post-HSCT chimerism analysis in predominantly Malay and Chinese patient-donor pairs. The commercially-available kits will also permit laboratories without extensive molecular biology capabilities to perform DNA typing in HSCT recipients.

Effect of ginseng saponins on beta-amyloid-suppressed acetylcholine release from rat hippocampal slices.

In view of the facts that ginseng has been shown to improve age-related memory deficits and beta-amyloid-related peptides have been suggested to play a significant role in memory degeneration in the elderly, the present study was carried out to examine the effect of various ginsenosides on beta-amyloid peptides-modulated acetylcholine (ACh) release, a key neurotransmitter in memory processing, from the hippocampal slices. Addition of beta-amyloid fragment(25 - 35) (0.01 - 1 microM) in the superfusion medium suppressed the K(+)-evoked [(3)H]-ACh release from the rat hippocampal slices in a concentration-related manner and about 40 % reduction in ACh outflow was observed when incubating with the highest concentration of an amyloid fragment (1 microM). Inclusion of the ginsenoside components Rb(1) (0.1 microM), but not Rg(1), caused a rightward shift of the concentration-response curve of beta-amyloid. The reversal of the beta-amyloid-inhibited ACh release by Rb(1) was not blocked by tetrodotoxin (1 microM) indicating that an interaction occurs at the cholinergic synapse. These results suggest that Rb(1) may elicit its anti-amnesic effect by minimizing the inhibitory effect of beta-amyloid peptides.

Complete genomic organization and promoter analysis of the round-spotted pufferfish JAK1, JAK2, JAK3, and TYK2 genes.

We have previously reported the isolation of the JAK1 gene from the round-spotted pufferfish. In the present study, we cloned and characterized genomic sequences encoding pufferfish JAK2, JAK3, and TYK2, which are other members of JAK family. To our knowledge, this is the first report to demonstrate the existence of four JAK genes in fish. All pufferfish JAK genes except JAK1 are composed of 24 exons; JAK1 has an additional exon. A comparison of the exon-intron organization of these genes revealed that the splice sites of JAK genes are nearly identical. In addition, all pufferfish JAK genes have one intron in the 5' untranslated region. Taken together, these data suggest that the pufferfish JAK genes may have evolved from a common ancestor. By 5' rapid amplification of cDNA ends and sequence analysis, we deduced the promoter regions for all JAK genes and found they do not contain typical TATA or CCAAT boxes but rather numerous other potential binding sites for transcription factors. Interestingly, the TYK2 gene is linked to CDC37 in a head-to-tail manner with a small intergenic region of 292 bp. Within this region, there are two potential binding sites for transcriptional factors such as c-Myb and NF-IL6. The putative promoter regions of all JAK genes were tested either in a carp CF cell line or in zebrafish embryos using CAT or lacZ as reporter genes. Both assays confirmed the transcriptional activities of these promoters in vitro and in vivo.

Effects of hetastarch and mannitol on prolonging survival in stable hypothermia in rats.

In rats, prolonged stable hypothermia ( approximately 24 h at body temperature of 19 degrees C) is characterized by a time-dependent increase in hematocrit, plasma osmolality, and red blood cell fragility and a decrease in plasma volume. These changes impede tissue microcirculation and could limit survival. As a countermeasure, we used plasma volume expanders of both long (hetastarch)- and short-lasting (mannitol) characteristics to improve microcirculation and hopefully hypothermia survival. Infusion of 6% hetastarch at hour 3 in hypothermia significantly (P < 0.05) enhanced survival over saline control (33.5 vs. 23.8 h); a significant delay in the increases of hematocrit and cell fragility was also observed compared with those in saline controls. Treating the animal with 6% hetastarch at hour 20 during hypothermia caused a similar but less-effective improvement in survival. In contrast, treating the rats with 6% mannitol at hour 3 or 20 during hypothermia failed to enhance survival over saline control, although transient improvement in plasma volume was observed. Our results indicate that by using a long-lasting volume expander, which tends to better maintain plasma volume and rheological parameters governing microcirculation than does saline or a short-lasting volume expander, hypothermia survival can be significantly improved.

Effect of ginseng saponins on cold tolerance in young and elderly rats.

Acute systemic injection of ginseng saponin (GS) significantly elevated both the total and maximum heat production in young rats (3-6 months) and improved their cold tolerance under severe cold (-10 degrees C under helium-oxygen). However, pretreating the animal with the optimal dose (10 mg/kg) of GS devoid of Rg1 and Rb1 failed to elicit any beneficial effect in improving the cold tolerance. Pretreating the animal with Rb1, but not Rg1, increased thermogenesis as well as cold tolerance in young rats. A similar beneficial effect in improving cold tolerance was also observed when old rats (26-28 months) were pretreated with the same doses of Rb1 (2.5 and 5.0 mg/kg). Our results indicate that Rb1 is the key ingredient in GS-mediated enhancement in thermogenic capacity and that both young and old rats can benefit from this treatment for enhanced cold tolerance.

Sentinel surveillance for enterovirus 71, Taiwan, 1998.

Outbreaks of enterovirus 71 have been reported around the world since 1969. The most recent outbreak occurred in Taiwan during April-July 1998. This hand, foot, and mouth disease epidemic was detected by a sentinel surveillance system in April at the beginning of the outbreak, and the public was alerted.

Effect of ginseng saponins on exercise performance in non-trained rats.

Short-term (4 days), but not acute, treatment with ginseng saponin (GS, 10 and 20 mg/kg/day) significantly prolonged the aerobic endurance of non-trained rats exercising at approximately 70% VO2max. Compared to the saline controls, GS treatment significantly increased the plasma free fatty acid (FFA) level and maintained plasma glucose level during exercise. Both the liver and skeletal muscle glycogen levels of the GS-treated rats were slightly higher than those of saline-treated controls after exhaustive exercise. These results indicate that GS enhances exercise endurance by altering fuel homeostasis during prolonged exercise, presumably by increasing FFA utilization in preference over glucose for cellular energy demands. To further search for the active components responsible for the ergogenic effect of GS, it was found that a GS preparation devoid of Rg1 and Rb1 failed, whereas injection of either Rg1 or Rb1 enhanced aerobic exercise performance. These results indicate that both Rg1 and Rb1 are key ingredients in GS-mediated enhancement in aerobic endurance.

Calcitriol enhancement of TPA-induced tumorigenic transformation is mediated through vitamin D receptor-dependent and -independent pathways.

We previously showed that 1alpha,25-dihydroxyvitamin D3, calcitriol, enhanced phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) induced tumorigenic transformation of mouse epidermal JB6 Cl41.5a cells. To determine if calcitriol regulates this enhancement through a nuclear vitamin D receptor (VDR)-dependent or -independent pathway, we used vitamin D analogs which induce biological responses by either of these mechanisms. In JB6 Cl41.5a cells, 1alpha,24-dihydroxy-22-ene-24-cyclopropyl-vitamin D3 (BT), which like calcitriol binds to VDR and regulates transcription, inhibited cell growth, stimulated expression of nonphosphorylated osteopontin (OPN), and enhanced TPA-induced anchorage-independent growth (AIG, an in vitro assay which highly correlates with tumorigenicity of these cells). 25-Hydroxy-16-ene-23-yne-vitamin D3 (AT), which stimulates calcium influx but has low affinity for VDR, had moderate effects on cell growth and expression of OPN. However, it enhanced TPA-induced tumorigenic transformation, though to a lesser extent than BT, thus suggesting that a VDR-independent mechanism is involved. Since 1alpha-hydroxylase activity was detected in JB6 cells, AT could be converted into 1alpha,25-dihydroxy-16-ene-23-yne-vitamin D3 (V), an analog which binds with high affinity to VDR, and could subsequently enhance TPA-induced AIG. To verify whether the VDR-independent pathway is involved in calcitriol enhancement of tumorigenic transformation, two additional VDR-independent analogs, 1alpha,25-dihydroxy-lumisterol3 (JN) and 24R,25-dihydroxyvitamin D3 (AS), were tested. The analog JN, which stimulates calcium transport and cannot be further hydroxylated at 1-carbon position, increased TPA-induced AIG, while AS, which inhibits calcium influx, did not. These studies suggest that a VDR-independent pathway, perhaps stimulation of calcium influx, and a VDR-dependent mechanism, which directly affects transcription, are involved in calcitriol's enhancement of TPA-induced tumorigenic transformation in JB6 Cl41.5a cells.