Scanning Tissue Oxygen Needle Probe.

A new device designed to scan oxygen partial pressure along a line in a biological tissue is described in this paper. The probe is housed in a stainless-steel needle. As opposed to other devices for oxygen scanning in tissue, the new probe does not require mechanical translation of the needle in the tissue. The probe includes an active sensing area along the needle shaft that can be scanned optically by an internal optical fiber. This feature allows for repeated scans of tissue oxygen along a line without translating the needle with respect to the tissue, thus avoiding tissue damage associated with needle motion. First, we describe the design of the device including its sensing mechanism, mechanical design, optical configuration, and signal processing. We then move on to describe the results of the device characterization and testing. Finally, we conclude by discussing possible applications of the device in research and in clinical diagnoses and treatment monitoring.

Elevated hepatic MDR3/ABCB4 is directly mediated by MiR-378a-5p in human obstructive cholestasis.

OBJECTIVE: The function of MDR3 is important in bile acid transport. The miRNAs can suppress the expression of gene through combining mRNA of target gene. The regulation about MDR3 mediated by FXR or PPARα in cholestasis is clear, but the mechanism through miRNA is hardly reported. We aimed to find out the miRNA, which could suppress MDR3 expression and the significance of this connection in cholestasis. PATIENTS AND METHODS: We measured hsa-miR-378a-5p expression level in liver tissues from 20 patients with cholestasis and 15 patients without cholestasis by quantitative PCR. We also tested the level of clinical features of the same group. HepG2 cell lines were performed experiments to discover the connection between hsa-miR-378a-5p and MDR3, including transient transfection, RNA and protein extraction, qPCR, Western blotting and luciferase reporter assay. RESULTS: A significant decrease of miR-378a-5p was observed in obstructive cholestasis patient liver tissues compared to control group. We also find that the miR-378a-5p expression is correlated to several clinical features, which are important biomarkers in cholestatic liver injury. Then we predicted that MDR3 may be the target gene of miR-378a-5p through miRanda v3.3a. We programed the transient transfection of mimics and inhibitor on HepG2 cell lines, and detected the mRNA and protein expression of MRP2, MRP3 and MDR3. The results suggested that miR-378a-5p could negatively regulate MDR3 expression in both mRNA and protein expression level, and this regulation is specific. We didn't find same regulation in MRP2 and MRP3. Dual luciferase assays proved this regulation is mediated by a direct binding between miR-378a-5p and CDS of MDR3. CONCLUSIONS: We found that hsa-miR-378a-5p expression was down-regulated in cholestatic liver tissues, compared to control liver tissues. Transient transfection and luciferase reporter assay in HepG2 cell lines results suggest that hsa-miR-378a-5p can directly combine MDR3 mRNA and suppress MDR3 protein expression. The down-regulated hsa-miR-378a-5p may cause a protective alteration through up-regulating MDR3 expression in cholestasis.

Diamine Functionalized Cubic Mesoporous Silica for Ibuprofen Controlled Delivery.

A diamine functionalized cubic mesostructured KIT-6 (N-KIT-6) has been prepared by post-synthetic method using calcined mesoporous KIT-6 with a diamine source, i.e., N-'[3-(tri methoxysilyl)- propyl]'ethylenediamine. The KIT-6 mesoporous silica used for N-KIT-6 was synthesized under weak acidic hydrothermal method using bitemplates, viz., Pluronic P123 and 1-butanol. The synthesized mesoporous materials, KIT-6 and N-KIT-6, have been characterized by the relevant instrumental techniques such as SAXS, N2 sorption isotherm, FT-IR, SEM, TEM and TGA to prove the standard mesoporous materials with the identification of diamine groups. The characterized mesoporous materials, KIT-6 and N-KIT-6, have been extensively used in the potential application of controlled drug delivery, where ibuprofen (IBU) employed as a model drug. The rate of IBU adsorption and release was monitored by UV vis-spectrometer. On the basis of the experimental results of controlled drug delivery system, the results of IBU adsorption and releasing rate in N-KIT-6 are higher than those of KIT-6 because of the higher hydrophobic nature as well as rich basic sites on the surface of inner pore wall silica.

The effects of surface roughness of composite resin on biofilm formation of Streptococcus mutans in the presence of saliva.

The purpose of this study was to investigate the effects of surface roughness of resin composite on biofilm formation of Streptococcus mutans in the presence of saliva. To provide uniform surface roughness on composites, disks were prepared by curing composite against 400-grit silicon carbide paper (SR400), 800-grit silicon carbide paper (SR800), or a glass slide (SRGlass). The surface roughness was examined using confocal laser microscopy. For biofilm formation, S. mutans was grown for 24 hours with each disk in a biofilm medium with either glucose or sucrose in the presence of fluid-phase or surface-adsorbed saliva. The adherent bacteria were quantified via enumeration of the total viable counts of bacteria. Biofilms were examined using scanning electron microscopy. This study showed that SR400 had deeper and larger, but fewer depressions than SR800. Compared to SRGlass and SR800, biofilm formation was significantly increased on SR400. In addition, the differences in the effect of surface roughness on the amount of biofilm formation were not significantly influenced by either the presence of saliva or the carbohydrate source. Considering that similar differences in surface roughness were observed between SR400 and SR800 and between SR800 and SRGlass, this study suggests that surface topography (size and depth of depressions) may play a more important role than surface roughness in biofilm formation of S. mutans .

Tumour oxygenation is increased by hyperthermia at mild temperatures. 1996.

The effects of hyperthermia on the oxygenation status in R3230 AC tumours of Fischer rats were measured using a polarographic oxygen electrode system. The median pO(2) in about 10 mm diameter tumours grown s.c. in the leg of rats was 3.7 +/- 0.3 mm Hg and it significantly increased upon heating at modest temperatures. For example, the tumour pO(2) measured within 10-15 min after heating for 30 min at 42.5 degrees C was about three-fold greater than that in the control tumours. About 62% of pO(2) values measured in control tumours were <5 mm Hg. After heating at 42.5 degrees C for 30 min, 37% of pO(2) values were <5 mm Hg. Such an increase in tumour oxygenation or reoxygenation of hypoxic cells appeared to result from an increase in tumour blood flow caused by the mild temperature hyperthermia. The presence of hypoxic cells in tumours is believed to be a major factor in limiting the effectiveness of radiotherapy, certain chemotherapy drugs and phototherapy. Hyperthermia at mild temperatures easily achievable with the use of presently available clinical hyperthermia devices may be an effective means to overcome the hypoxic protection in the treatment of human tumours.

Toxicity of repeated intravenous injection of gene therapeutics for X-CGD in mice.

We made gene therapeutics for X-chronic granulomatous disease (CGD) by transducing murine bone marrow-derived stem cells with MT-gp91 retrovirus and evaluated possible toxicity in mice as a prerequisite for human clinical trials. Male C57BL/6 mice were injected intravenously with gene therapeutics for X-CGD twice at an interval of two weeks at 5 x 10(7) cells/kg and sacrificed 2 weeks after the last administration. Significant changes noted in gene therapeutics for X-CGD-treated animals were an increase in white blood cell counts and a slight decrease in albumin/globulin ratio. The red pulp hyperplasia in the spleen accompanied with an increase in organ weight was considered to result from the accumulation of gene therapeutics for X-CGD, bone marrow-derived stem cells, in the spleen. No anti-gp91 antibody was detected in the sera collected from the animals treated with gene therapeutics for X-CGD. No integration of gp91 DNA from retroviral vector was detected in chromosomal DNA of gonads in animals dosed with the test substance, indicating no potential of genomic integration. In conclusion, the repeated dose of gene therapeutics for X-CGD exerted no toxicity. The splenic red pulp hyperplasia and the increase observed in white blood cell counts and in spleen weights were considered as pharmacological changes induced by the treatment.

COX-2 inhibitor celecoxib prevents chronic morphine-induced promotion of angiogenesis, tumour growth, metastasis and mortality, without compromising analgesia.

Morphine and its congener opioids are the main therapy for severe pain in cancer. However, chronic morphine treatment stimulates angiogenesis and tumour growth in mice. We examined if celecoxib (a cyclooxygenase-2 (COX-2) inhibitor) prevents morphine-induced tumour growth without compromising analgesia. The effect of chronic treatment with celecoxib (by gavage) and/or morphine (subcutaneously), or PBS on tumour prostaglandin E(2) (PGE(2)), COX-2, angiogenesis, tumour growth, metastasis, pain behaviour and survival was determined in a highly invasive SCK breast cancer model in A/J mice. Two weeks of chronic morphine treatment at clinically relevant doses stimulates COX-2 and PGE(2) (4.5-fold compared to vehicle alone) and angiogenesis in breast tumours in mice. This is accompanied by increased tumour weight ( approximately 35%) and increased metastasis and reduced survival. Co-administration of celecoxib prevents these morphine-induced effects. In addition, morphine and celecoxib together provided better analgesia than either agent alone. Celecoxib prevents morphine-induced stimulation of COX-2, PGE(2), angiogenesis, tumour growth, metastasis and mortality without compromising analgesia in a murine breast cancer model. In fact, the combination provided significantly better analgesia than with morphine or celecoxib alone. Clinical trials of this combination for analgesia in chronic and severe pain in cancer are warranted.

Genomic organization of the region spanning D14Mit262 and D14Mit86 on mouse chromosome 14 and exclusion of Adam28 and Adamdec1 as the cataract-causing gene, lr2.

Mice with recessive cataract, CXSD, show the first clinical symptoms of cataract at five weeks, with complete penetrance. We previously localized the cataract-causing lens rupture 2 gene (lr2) to mouse chromosome 14. In the process of positional cloning of the lr2 gene, we determined the genomic organization of the critical region, defined by D14Mit262 and D14Mit86, and compared it to recently published map information. In addition, mutational analysis using reverse transcription polymerase chain reaction (RT-PCR) followed by direct sequencing as well as quantitative realtime PCR (RQ-PCR) was performed to investigate Adam28 and Adamdec1 as lr2 candidate genes in this study. There was no mutation cosegregating with the phenotype of CXSD mice, which excluded these genes as the lr2 gene. Identification of more transcripts from this region and their mutation analyses are required to isolate the lr2 gene.

Implications of increased tumor blood flow and oxygenation caused by mild temperature hyperthermia in tumor treatment.

In many past clinical studies in which hyperthermia enhanced the efficacy of radiotherapy, the tumor temperatures could be raised only to 40-42 degrees C range in most cases. The heat-induced cell death, cellular radiosensitization, and vascular damage induced by such mild temperature hyperthermia (MTH) are likely to be insignificant despite the increased response of tumors to radiotherapy. Heating rodent tumors at 40-42 degrees C was found to cause an enduring increase in blood flow and oxygenation in the tumors. Recent studies with canine soft tissue sarcoma and human tumor clinical studies also demonstrated that MTH improves tumor oxygenation, and enhances response of the tumors to radiotherapy or chemoradiotherapy. The increased blood flow and vascular permeability caused by MTH may also improve the delivery of various therapeutic agents such as chemotherapy drugs, immunotherapeutic agents and genetic constructs for gene therapy to tumor cells. MTH as a means to potentiate the efficacy of radiotherapy and others warrants further investigation.

Quercetin inhibits LPS-induced delay in spontaneous apoptosis and activation of neutrophils.

OBJECTIVE: To investigate the effects of quercetin, an herbal flavonoid, on LPS-induced delay in spontaneous apoptosis, adhesion molecules (CD62L, CD11b/CD18) expression of neutrophils, and superoxide (O(2)(-)) generation by LPS-primed fMLP-induced human neutrophils. METHODS: Neutrophils were incubated in the presence or absence of lipopolysaccharide (LPS) at a final concentration of 1 microg/ml for 24 hours. Some wells with neutrophils were pre-treated with quecetin at the final concentration ranging from 0-100 microM for 30 min and then 1 microg/ml LPS was added into the cultures for 24 hours. The apoptosis of neutrophils was evaluated by flowcytometry analysis of propidum iodide (PI)-staining of the nuclei and annexin V staining of phosphatidylserine (PS) in the cell membrane. Agarose gel electrophoresis of low molecular weight DNA was performed to analyze DNA fragmentation. The effects of quercetin on adhesion molecules were detected by using flowcytometry analysis. The generation of O(2) (-) by LPS-primed fMLP-induced neutrophils was determined by reduced cytochrome c assay. RESULTS: LPS markedly inhibited the spontaneous apoptosis of neutrophils, but the inhibitory effect was abrogated after the pre-treatment of neutrophils with quercetin (approximately 40 microM) for 30 min. Quercetin (40 microM) also prevented LPS-induced down-regulation of CD62L expression, up-regulation of CD11b/CD18 expression, and O(2) (-) generation by fMLP-induced neutrophils. CONCLUSION: As one of the pro-inflammatory factors, LPS aggravates inflammation through priming neutrophils to synthesize/release cytotoxic contents and prolonging functional lifespan of neutrophils by delaying the spontaneous apoptosis. Thus, our data suggest to us that quercetin might decrease the susceptibility of neutrophils to pro-inflammatory factors (e. g. LPS), which could partially explain the anti-inflammatory mechanisms of quercetin.

Physiological mechanisms underlying heat-induced radiosensitization.

The objective of this review is to evaluate hyperthermia related changes in tumor physiologic parameters and their relevance for tumor radiosensitization with particular emphases on tumor oxygenation. Elevation of temperature above the physiological level causes changes in blood flow, vascular permeability, metabolism, and tumor oxygenation. These changes in addition to the cellular effects such as direct cytotoxicity, inhibition of potentially lethal damage and sublethal damage repair, have an important influence on the efficacy of radiotherapy. There is now clear evidence that in a variety of rodent and canine, as well as human tumors, the changes in tumor oxygenation status caused by hyperthermia are temperature dependent and this relationship may greatly influence the response of tumors to thermoradiotherapy. The improvement of tumor oxygenation after mild hyperthermia, which often lasts for as long as 24-48 h after heating, may increase the likelihood of a positive response of tumors to radiation therapy. Furthermore, the activity of some chemotherapy drugs is also oxygen dependent, therefore, the heat-induced increase in tumor oxygenation may significantly increase the effectiveness of thermoradiotherapy in combination with certain chemotherapy drugs. Further investigations remain to be conducted to obtain clearer insights into the relationship between thermal parameters, oxygenation and response of human tumors to hyperthermia in combination with radiotherapy and/or chemotherapy.

Arsenic trioxide induces selective tumour vascular damage via oxidative stress and increases thermosensitivity of tumours.

It has previously been found that the anti-leukaemia agent Arsenic Trioxide (ATO) causes vascular shutdown in solid tumours and markedly sensitizes tumours to hyperthermia. The present study was designed to evaluate the mechanism of action and dose-dependence of ATO-induced thermosensitization in FSaII and SCK murine tumours. The role of oxidative stress was studied by observing ATO-induced vascular shutdown in vivo and ATO-induced endothelial cell adhesion molecule expression in vitro in the presence or absence of an anti-oxidant. It was found that a dose as low as 2 mg/kg ATO impaired vascular function, as estimated by 86Rb uptake, in the tumour. The degree of tumour growth delay induced by 1 h of hyperthermia at 42.5 degrees C, applied 2 h after ATO injection, was proportional to the dose of ATO administered. In addition, it was found that ATO can directly thermosensitize tumour cells in vitro. The development of massive tissue necrosis in the tumour was observed in the days after treatment, especially with the combination of ATO and heating. ATO-induced adhesion molecule expression in vitro was abolished when the anti-oxidant n-acetyl-cysteine (NAC) was introduced prior to exposure, while the addition of NAC in vivo partially blocked ATO-induced vascular shutdown. These results suggest that the expression of adhesion molecules by the vasculature due to oxidative stress contribute to the ATO-induced selective tumour vascular effects observed and that the clinical use of ATO to increase tumour thermosensitivity via direct cellular and vascular effects appears feasible.

Effect of glycolic acid on UVB-induced skin damage and inflammation in guinea pigs.

OBJECTIVES: Recently the use of glycolic-acid-containing cosmetics has received increased public interest in their supposed ability to reduce wrinkles, roughness, age spots and other skin damage. However, the safety of such products when used excessively or chronically, especially by photosensitive people, is being questioned. The purpose of this study was to examine the effects of glycolic acid alone or in combination with UVB on skin damage and inflammatory response. METHOD: Guinea pigs were treated with glycolic acid (from 1 to 7 mg/cm(2)) alone or in combination with UVB (0.4 or 3 J/cm(2)) for 14 days. Skin damage was evaluated by scoring the skin irritation value by the method of Draize and by histopathological observations. Cyclooxygenase 2 (COX-2) expression and prostaglandin E(2) (PGE(2)) production were also assessed. RESULTS: Glycolic acid caused an increase in the level of skin damage in a dose- and time-dependent manner. Lower doses (1 and 3 mg/cm(2)) of glycolic acid mostly caused erythema and eschar, and these consequently formed scales, whereas higher doses (5 and 7 mg/cm(2)) of glycolic acid caused redness, edema and necrotic ulceration. Glycolic acid also increased the thickness of the epidermal layer, reduced the organization of the stratum corneum and eventually destroyed some parts of the epidermal layer at 7 mg/cm(2). UVB (0.4 and 3 J/cm(2)) caused redness and edema as well as reduced the integrity of the stratum corneum. Glycolic acid enhanced the UVB-induced skin damage. The magnitude of the damage caused by combined UVB and glycolic acid treatment was much greater than that caused by glycolic acid or UVB alone. Moreover, partial destruction of the epidermal layer was observed in skin treated with 3 J/cm(2) UVB and 3 mg/cm(2) glycolic acid. However, glycolic acid did not change the basal and UVB-induced PGE(2) production and COX-2 protein expression. CONCLUSION: These results show that glycolic acid causes skin damage in a dose- and time-dependent manner and that it enhances UVB-induced skin damage without accompanying PGE(2) production or COX-2 protein expression. Therefore, caution should be exercised by those using glycolic acid on a chronic basis or excessively. Moreover, those with photosensitive skins and those more exposed to the sun should be particularly careful.

Inconsistent association of esophageal symptoms, psychometric abnormalities and dysmotility.

OBJECTIVES: The aim of this study was to characterize the psychometric profiles of symptomatic patients with abnormal esophageal motility and symptomatic patients with normal manometric findings compared to asymptomatic controls. METHODS: A total of 113 patients with abnormal esophageal motility (7 achalasia, 8 diffuse esophageal spasm, 27 nutcracker esophagus, 37 hypertensive lower esophageal sphincter, 21 hypotensive peristalsis, 13 failed peristalsis), 23 symptomatic controls with similar esophageal symptoms but normal manometry, and 27 asymptomatic controls were enrolled. Validated questionnaires assessing depression (Beck Depression Inventory), anxiety (Spielberger State Anxiety Inventory or Trait Anxiety Inventory), and somatization (Psychosomatic Symptom Checklist) were administered to all subjects. RESULTS: Patients with both esophageal symptoms and either hypertensive lower esophageal sphincter, nutcracker esophagus, or hypotensive contractions exhibited increased somatization, acute anxiety, or depression compared to asymptomatic controls but not compared to symptomatic controls. On the other hand, the psychometric profiles of patients with achalasia and diffuse esophageal spasm were strikingly normal. Among esophageal symptoms, chest pain was closely correlated with psychometric abnormalities. CONCLUSIONS: The esophageal symptoms of patients with abnormal esophageal motility may relate to the underlying psychological abnormalities, independent of manometric abnormalities.

Endoscopic removal of sharp foreign bodies impacted in the esophagus.

BACKGROUND AND STUDY AIMS: Impacted sharp foreign bodies in the esophagus can be very difficult to manage. When attempts are made to remove such objects inappropriately, life-threatening complications such as perforation can occur. The aim of this study was to evaluate the safety and efficacy of endoscopic removal of impacted sharp foreign bodies in the esophagus using proximal dilatation with an oral side balloon or transparent cap. PATIENTS AND METHODS: A total of 22 patients (10 men, 12 women) with impacted sharp foreign bodies in the esophagus underwent endoscopic extraction. The following technique was successfully performed at our hospital. An oral side balloon (Top Co., Japan) for esophageal variceal sclerotherapy was attached to the distal part of the endoscope. With the patient under local anesthesia, the endoscope was inserted as far as the proximal part of the esophageal foreign body. The oral side balloon was then gradually inflated. Dilatation of the proximal part of the esophagus made it possible to release the impacted sharp foreign body from the esophageal wall. A transparent cap was used for foreign bodies in the upper esophagus when there were difficulties with the oral side balloon. RESULTS: The types of foreign body removed were fish bones (n = 9), press-through packages (n = 8), chicken bones (n =3), dentures (n = 1), and a wrist watch (n = 1). Endoscopic removal was successful in all but one of the cases, in which a fish bone had to be extracted surgically. CONCLUSIONS: The proximal dilatation method using an oral side balloon or transparent cap is safe and effective in removing sharp foreign bodies from the esophagus, avoiding surgery and possible perforation.

Enhancement of cell killing by induction of apoptosis after treatment with mild hyperthermia at 42 degrees C and cisplatin.

Ohtsubo, T., Igawa, H., Saito, T., Matsumoto, H., Park, H. J., Song, C. W., Kano, E. and Saito, H. Enhancement of Cell Killing by Induction of Apoptosis after Treatment with Mild Hyperthermia at 42 degrees C and Cisplatin. Radiat. Res. 156, 103-109 (2001). We examined the interactive effects of cisplatin (1.0 microg/ml) combined with hyperthermia on cell killing and on the induction of apoptosis in IMC-3 human maxillary carcinoma cells. The cytotoxic effects of hyperthermia on IMC-3 cells at 44 degrees C were greater than at 42 degrees C, as has been reported for many other cells. The induction of apoptosis, DNA fragmentation and poly(ADP-ribose) polymerase cleavage were greater after hyperthermia at 44 degrees C for 30 min compared with treatment at 42 degrees C for 105 min, even though both of these heat doses were isoeffective in reducing cell survival to 50%. Treatment with cisplatin at 37 degrees C for up to 120 min did not result in cytotoxicity or the induction of apoptosis. The enhancement ratio for treatment with cisplatin at 42 degrees C was greater than that at 44 degrees C. More apoptosis was induced after the treatment with cisplatin at 42 degrees C compared to treatment with cisplatin at 44 degrees C. Taking these findings together, the combination of cisplatin and hyperthermia at 42 degrees C appeared to be more effective than cisplatin with hyperthermia at 44 degrees C for the induction of apoptosis in IMC-3 cells.

Prevalence of endoscopic reflux esophagitis among Koreans.

BACKGROUND AND AIM: Although reflux esophagitis is believed to be common in the Western population, very few epidemiologic data on reflux esophagitis in Koreans are available. The aims of this study were to evaluate the prevalence of endoscopic reflux esophagitis in patients who came for a physical check-up at Korea University Hospital, and to study the relationship between various factors relevant to reflux disease. METHODS: This study was carried out prospectively on 7,015 patients who received an esophagogastroduodenoscopy from September 1996 to December 1997. Most of the patients were free of symptoms and had come for their self-paid check-up. RESULTS: The overall prevalence of reflux esophagitis was 3.4%, and most of the patients had a mild degree of esophagitis representing grade 1 in 98.3% and grade 2 in 1.7%. The male: female ratio for esophagitis was 7 : 1, and the body mass index (BMI) was significantly higher in patients with reflux esophagitis. A hiatal hernia was found in 166 patients with esophagitis (68.6%), but only in 9.2% patients without esophagitis (P < 0.05). Smoking and alcohol consumption were associated with the development of reflux esophagitis (P < 0.05). CONCLUSIONS: The prevalence of endoscopic reflux esophagitis among Koreans is 3.4%, and most of the patients had a mild grade esophagitis. Smoking, alcohol consumption, the presence of a hiatal hernia and a higher BMI are associated with the development of reflux esophagitis.

Lead treatment in vitro at early developmental stage of bone marrow-derived macrophages enhances NO production through IL-1beta and IL-6 but not TNF-alpha.

Lead (Pb(2+)) is known to decrease or block nitric oxide (NO) production by mature macrophages (mphi). Bone marrow cells were treated with various doses of lead in vitro and the morphological and functional changes were observed. Bone marrow cells were treated with various doses of lead (1, 10, 20 and 50 microM) at the start of culture with mphi growth factor (CSF-1), and after 6-7 days of culture, the resultant mphi (bone marrow-derived mphi, BMDM) showed decreased NO production. Unexpectedly, BMDM from the lowest does of lead treatment (1.0 microM) showed increased NO production. The increased NO production was due to increased expression of the iNOS gene and concurrent enhanced transcript levels of proinflammatory cytokines such as IL-1beta and IL-6, but not TNF-alpha. Lead treatment on mature BMDM showed decreased NO production in a dose-dependent manner. These results suggest that a low dose of lead affects developmental characteristics of BMDM through different proinflammatory cytokines, and the lead effects on precursor cells of mphi and mature mphi are different.

Acidic environment modifies heat- or radiation-induced apoptosis in human maxillary cancer cells.

PURPOSE: The effects of hyperthermia or irradiation on cell killing and induction of apoptosis were evaluated using human maxillary carcinoma IMC-3 cells and low pH (pH 6.8) adapted cells (IMC-3-pH). METHODS AND MATERIALS: Cellular heat-sensitivity or radiosensitivity was determined using the clonogenic assay. Apoptosis was assessed on the basis of a flow cytometric determination of the DNA content, DNA fragmentation, and poly(ADP-ribose)polymerase cleavage. RESULTS: When IMC-3 cells or IMC-3-pH cells were exposed to heat at 44 degrees C in pH 6.8 medium, an increase in thermosensitivity was observed compared with when the IMC-3 cells were exposed to heat at 44 degrees C in pH 7.4 medium. However, the selective reduction in survival was not observed after irradiation. In IMC-3 cells, apoptosis after heating at 44 degrees C for 60 min in pH 7.4 medium occurred earlier than that after 8 Gy irradiation, although both thermal and irradiated doses decreased the cell count to 10%. The degree of apoptosis after heating at pH 6.8 in IMC-3 cells or IMC-3-pH cells was greater than that at pH 7.4 in IMC-3 cells. However, the degree of apoptosis after 8 Gy irradiation at pH 6.8 in IMC-3 cells or IMC-3-pH cells was smaller than that at pH 7.4 in IMC-3 cells. CONCLUSION: Hyperthermia treatment is more effective at inducing apoptosis than radiation is in tumors that contain a population of low pH adapted cells.