The marine dinoflagellate, Gonyaulax polyedra emits light in a reaction involving the enzymatic oxidation of its tetrapyrrole luciferin by molecular oxygen; its luciferase (LCF) single chain has an estimated molecular mass of 130 kDa, and exhibits a circadian rhythm in its activity. A cDNA expression library in the lambda ZAPII vector was constructed from the polyadenylated RNA isolated from the Gonyaulax cells during the early night phase, the time at which LCF synthesis is believed to be greatest. Of the approx. 1.2 . 10(5) phages from the library screened with antibody against Gonyaulax LCF, 13 positive plaques were obtained. The nucleotide sequences of two of the larger inserts (2.4 kb and 1.6 kb in length), both carrying the poly(A) tail, were determined and found to be identical in the overlapping region. When expressed in Escherichia coli, both cDNA clones produced active luciferase. A Northern hybridization using the cDNA as a probe showed that the length of the lcf mRNA is approx. 4.1 kb, sufficiently long to encode the 130 kDa LCF. Analyses of polymerase chain reaction products, prepared using both the cloned cDNA and Gonyaulax chromosomal DNA as templates, indicated that the cloned region of the luciferase gene does not carry any introns. This represents the first dinoflagellate luciferase to be cloned and sequenced; its deduced amino acid sequence bears no significant homologies with that of any other luciferase, or any other sequence in the data base.

Brain-derived neurotrophic factor (BDNF) is a neurotrophin that influences neuronal plasticity throughout life. Emergence from a vegetative state (VS) after a traumatic brain injury (TBI) implies that the brain undergoes plastic changes. A common polymorphism in the BDNF gene--BDNF Val66Met (referred to herein as BDNF(Met))--impairs cognitive function in healthy subjects. The aim of this study was to determine whether the BDNF(Met) polymorphism plays a role in the recovery of consciousness and cognitive functions in patients in a VS after a TBI. Fifty-three patients in a VS 1 month after a TBI were included in the study and genotyped for the BDNF(Met) polymorphism. Scores of levels of cognitive functioning (LCF) at 1, 3, 6, and 12 months post-TBI were retrospectively compared in patients without (Val group), and with (Met group), the BDNF(Met) polymorphism. The BDNF(Met) polymorphism was detected in 20 out of the 53 patients. The mean LCF scores in the Val and Met groups were 1.6±0.5 and 1.4±0.5 at 1 month, 2.3±0.7 and 2.5±1.2 at 3 months, 3.3±1.7 and 3.5±1.7 at 6 months, and 4±1.9 and 3.9±1.8 at 12 months, respectively (p>0.05). The percentages of patients in the Val and Met groups who emerged from the VS were 36.4% and 30% at 3 months, 66.3% and 70% at 6 months, and 70% and 87.5% at 12 months (p>0.05), respectively. These findings provide evidence that the BDNF(Met) polymorphism is not involved in cognitive improvement in patients with a VS following TBI. Future studies should focus on the role of other BDNF polymorphisms in the recovery from a VS.

The aim of this study was to characterize human isolates of Lactobacillus species for their capacity to interfere with the growth of different strains of Candida species in vitro in the search for a potential probiotic. Growth inhibition of Candida species was screened using an agar-overlay method. Inhibiting strains were selected to assay the effect of a cell-free Lactobacillus culture filtrate (LCF) on the growth of isolates of Candida albicans and Candida glabrata. A total of 126 human Lactobacillus isolates was investigated. Eighteen isolates significantly inhibited the growth of C. albicans on agar. The LCF of one of these strains showed strong inhibition of both C. albicans and C. glabrata. This strain was genetically identified as Lactobacillus fermentum and designated L. fermentum Ess-1. Further tests to evaluate the probiotic potential of this strain indicated that L. fermentum Ess-1 strain is a promising probiotic for use in clinical trials to treat and prevent vulvo-vaginal candidiasis.

Recent research suggests that an attentional bias toward threat may play a causal role in obsessive-compulsive disorder (OCD) with contamination concerns. However, the attentional components involved in this bias, as well as its behavioral correlates, remain unclear. In the present study, eye movements were recorded in individuals high and low in contamination fear (HCF, LCF, respectively) during 30-s exposures to stimulus arrays containing contamination threat, general threat, pleasant, and neutral images. HCF individuals oriented gaze toward contamination threat more often than LCF individuals in initial fixations, and this bias mediated group differences in responding to a behavioral challenge in a public restroom. No group differences were found in the maintenance of gaze on contamination threat, both in terms of initial gaze encounters, as well as gaze duration over time. However, the HCF group made shorter fixations on contamination threat relative to other image types. The implications of these findings for further delineating the nature and function of attentional biases in contamination-based OCD are discussed.

Mitogens and antigens have been the traditional ligands for activating lymphocytes in vitro for the elaboration of lymphokines. Recently, histamine, by interaction with histamine-type 2 receptors on T lymphocytes, has been found to induce the production of one lymphokine, histamine-induced suppressor factor (HSF), that inhibits lymphocyte proliferation and lymphokine production in vitro. Because the biologic effects of HSF appear to be confined to alterations in lymphocyte function, we assessed the ability of soluble products of histamine-stimulated human blood mononuclear cells to affect another lymphocyte function, motility. Utilizing a modified Boyden chamber assay to assess lymphocyte migration, we identified chemoattractant activity for human blood and rat splenic T lymphocytes in histamine-induced mononuclear cell supernatants. No neutrophil or monocyte chemoattractant activity was present. Sephadex G-100 gel filtration of histamine-induced supernatants showed the lymphotactic activity eluted with a 56,000 m.w. This activity was cationic as determined by its elution pattern from a Sephadex QAE anion exchange matrix with a single pl of 9.0 to 9.4 determined by isoelectric focusing in sucrose. Its biologic activity is predominantly chemokinetic in nature, is stable to heating at 56 degrees C for 30 min, but is sensitive to the effects of trypsin and neuraminidase. These physicochemical and functional characteristics establish it as identical to a recently described concanavalin A-induced (Con A) lymphotactic lymphokine (LCF). Mononuclear cells that did not adhere to a histamine affinity matrix were unable to produce LCF when subsequently stimulated with histamine or Con A. Mononuclear cells incubated with histamine and diphenhydramine produced LCF; the addition of cimetidine eliminated LCF production. In fact, supernatants from cells incubated with histamine and cimetidine significantly inhibited lymphocyte migration, a phenomenon explainable by the two regions of lymphocyte migration inhibitory activity that were present in the Sephadex G-100 chromatography of crude histamine-induced supernatants. These data suggest that a subset of lymphocytes defined by the presence of histamine-type 2 receptors is capable of producing LCF while cells that bear histamine-type 1 receptors produce lymphocyte migration inhibitory activity.

OBJECTIVE

The aim of this study was to investigate the efficacy of artemether-lumefantrine in treating uncomplicated Plasmodium falciparum malaria in four sentinel areas in Sudan with different malaria transmission (Damazin, Sinnar, and Kosti in the north, and Juba in the south).

METHODS

World Health Organization protocol for assessing antimalarial drug efficacy in treating uncomplicated P. falciparum malaria was employed. A total of 2,139 patients were screened, and 771 had P. falciparum monoinfection. Only 291 met the enrollment criteria and gave written consent to be recruited in the study. Patients were treated with artemether-lumefantrine tablets in a six-dose regimen calculated according to body weight. Tablets were given at 0, 8, 24, 36, 48, and 60 h. Patients were followed up for 28 days.

RESULTS

A total of 291 patients were recruited to the study, of whom ten [3.4; 95% confidence interval (CI):1.8-6.4%] patients showed early treatment failure (ETF) or late clinical failure (LCF) and were excluded from further follow-up. Of the remaining 281 patients, 276 (98.2%; 95% CI: 95.7-99.3%) completed the 28-day follow-up. Of these, 274 (99.3%; 95% CI: 97.1-99.9%) had adequate clinical and parasitological response (ACPR), and two (0.7%; 95% CI: 0.13-2.9%) showed late parasitological failure (LPF) at days 21 and 28. The overall mean +/- standard deviation (SD) of parasitemia and fever clearance times were 36.4 (23.7) h and 34.6 (19.2) h, respectively. Mild and reversible adverse effects were reported by 11 patients (3.8%; CI: 2.0- 7.0%) and were relieved without the need for termination of drug therapy or supportive treatment.

CONCLUSIONS

Our findings showed that artemether-lumefantrine was an effective and safe drug for treating uncomplicated P. falciparum malaria in northern and southern Sudan.

OBJECTIVE

To test the hypothesis that infection control practices can prevent the spread of vancomycin-resistant enterococci (VRE) to residents of a long-term care facility (LCF) from an affiliated acute care facility with a high endemic rate of colonization.

METHODS

Point prevalence study of the rate of rectal colonization.

METHODS

A state-supported veterans nursing home and an acute care veterans hospital.

METHODS

Residents in a state veterans home.

METHODS

Identification of patients with rectal colonization by VRE before transfer to the state veterans home, contact isolation for colonized veterans, use of oral bacitracin to eliminate colonization.

METHODS

Rectal swab and culture for VRE, review of clinical records and recording of presumptive risk factors for VRE colonization. The risk factors were age, gender, length of stay at nursing home, treatment with vancomycin or oral antibiotics, prior hospitalization at the acute care facility during the prior year, use of indwelling urethral catheters, presence of diarrhea, and fecal or urinary incontinence.

RESULTS

Sixty-nine of 200 residents were cultured in the first study (1996) and 130 of 230 residents were cultured in the second study (1998). Residents who consented to culture differed from those who did not only with regards to gender (2 vs 7, P = .012). In neither study were any residents found to be colonized with VRE who had not already been identified as positive on admission.

CONCLUSIONS

Adherence to infection control practices by the patient care staff of the LTCF was associated with the absence of transmission of VRE colonization among its residents. The presence of rectal colonization with VRE in an acute care patient should not be a barrier to acceptance in a nursing home.

We describe the practical issues and the methodological procedures that must be carried out to construct and use QSAR models for predicting localization of probes in single cells. We address first the determination of probe factors starting with a consideration of the chemical nature of probe molecules present. What is their identity? Do new compounds arise in incubation media or intracellularly? For each probe, how many distinct chemical species are present? For each probe species, the derivation of the following numerical structure parameters, or descriptors, is set out with worked examples of electric charge and acid/base strength (Z and pKa); hydrophilicity/lipophilicity (log P); amphiphilicity (AI and HGH); conjugated bond number and largest conjugated fragment (CBN and LCF); width and length (W and L); and molecular and ionic weights, head group size and substituent bulk (MW, IW, HGS and SB). Next, protocol factors are specified by focusing separately on the mode of introduction of the probe to the cells, other application phenomena, and factors that influence directly observations of outcomes. Cell factors then are specified by considering separately structural and functional aspects. The next step is to select appropriate QSAR models and to integrate probe, protocol and cell factors to predict the interactions of the probe with the cell. Finally, we use an extended case example to explore the intracellular localization of certain photodynamic therapy dyes to illustrate these procedures.

Determining the effectiveness of in situ immobilization for P-amended, Pb-contaminated soils has typically relied on non-spectroscopic methods. However in recent years, these methods have come under scrutiny due to technical and unforeseen error issues. In this study, we analyzed 18 soil samples via X-ray diffraction (XRD), selective sequential extraction (SSE), and a physiologically based extraction test (PBET). The data were compared against each other and to previous data collected for the soil samples employing X-ray absorption fine structure spectroscopy coupled with linear combination fitting (XAFS-LCF), which spectroscopically speciates and quantifies the major Pb species in the samples. It was observed that XRD was incapable of detecting pyromorphite, the hopeful endpoint of the immobilization strategy for reduced Pb bioavailability in our studies. Further, the SSE and PBET extraction methods demonstrated an increase of recalcitrant Pb forms in comparison to the XAFS-LCF results suggesting that SSE and PBET methods induced the precipitation of pyromorphite during the extraction procedures. The theme of this paper illustrates the experimental concerns of several commonly employed methods to investigate immobilization strategies of amended, metal-contaminated systems which may not be in true equilibrium. We conclude that appropriate application of spectroscopic methods provides more conclusive and accurate results in environmental systems (i.e., Pb, Zn, Cd, etc.) examining P-induced immobilization.

OBJECTIVE

To evaluate the bending fatigue lifetime of nickel-titanium alloy (NiTi) and stainless steel (SS) endodontic files using finite element analysis.

METHODS

The strain-life approach was adopted and two theoretical geometry profiles, the triangular (TR) and the square cross-sections, were considered. Both low-cycle fatigue (LCF) lifetime and high-cycle fatigue (HCF) lifetime were evaluated.

RESULTS

The bending fatigue behaviour was affected by the material property and the cross-sectional configuration of the instrument. Both the cross-section factor and material property had a substantial impact on fatigue lifetime. The NiTi material and TR geometry profiles were associated with better fatigue resistance than that of SS and square cross-sections.

CONCLUSIONS

Within the limitations of this study, finite element models were established for endodontic files to prejudge their fatigue lifetime, a tool that would be useful for dentist to prevent premature fatigue fracture of endodontic files.

Both pulsed and continuous applications of the RNA polymerase II inhibitor thiolutin cause a dramatic but reversible loss of bioluminescence and its overt rhythmicity in cells of the dinoflagellate Lingulodinium polyedrum (formerly Gonyaulax polyedra). Such cells remain alive, and the rhythm resumes after an interval, the length of which depends on the concentration of thiolutin used. The period and phase of the resumed rhythm were not systematically altered following such treatments, and the effects were not different at different circadian phases. For three different genes, luciferin binding protein (lbp), luciferase (lcf), and glyceraldehyde-3-phosphate dehydrogenase (gapdh), which are circadian-regulated at the level of translation, the amounts of their mRNAs were determined by Northern blots for times up to 12.5 h following the addition of 1.5 microM thiolutin. Consistent with previous reports that their abundances do not change with circadian time, their levels remained high for several hours after thiolutin addition, but then did diminish.

Recent studies on the speciation of Zn in contaminated soils confirmed the formation of Zn-layered double hydroxide (LDH) and Zn-phyllosilicate phases. However, no information on the kinetics of the formation of those phases under field conditions is currently available. In the present study, the transformation of Zn in a field soil artificially contaminated with ZnO containing filter dust from a brass foundry was monitored during 4 years using extended X-ray absorption fine structure (EXAFS) spectroscopy. Soil sections were studied by micro-X-ray fluorescence (micro-XRF) and micro-EXAFS spectroscopy. EXAFS spectra were analyzed by principal component analysis (PCA) and linear combination fitting (LCF). The results show that ZnO dissolved within 9 months and that half of the total Zn reprecipitated. The precipitate was mainly of the Zn-LDH type (>75%). Only a minor fraction (<25%) may be of Zn-phyllosilicate type. The remaining Zn was adsorbed to soil organic and inorganic particles. No significant changes in Zn speciation occurred from 9 to 47 months after the contamination. Thermodynamic calculations show that both Zn-LDH and Zn-phyllosilicate may form in the presence of ZnO but that the formation of Zn-phyllosilicate would be thermodynamically favored. Thus, the dominance of Zn-LDH found by spectroscopy suggests that the formation of the Zn precipitates was not solely controlled bythermodynamics but also contained a kinetic component. The rate-limiting step could be the supply of Al and Si from soil minerals to the Zn-rich solutions around dissolving ZnO grains.

Lymph node cells from guinea pigs sensitized by BCG elaborated two chemotactic factors for lymphocytes when stimulated with purified protein derivative in vitro 4 weeks after immunization. Production of the factors occurred within 24 h of incubation. The two factors were termed alpha- and beta-lymphocyte chemotactic factors (alpha-LCF, beta-LCF), relative to their elution order on gel chromatography. The molecular weights were about 160,000 and 27,000, respectively. alpha-LCF was labile to heating at 56 degrees C for 30 min. Both factors were sensitive to digestion by trypsin, and were effective for T cells but not for B cells.

To unravel the contributions of corneal tissue in endocular inflammation, we examined the capability of corneal endothelial cells (CECs) to release leukocyte chemotactic factors (LCFs) following injury induced by the leukocyte product hydrogen peroxide. Hydrogen peroxide (H2O2) was chemically generated by the interactions of glucose and glucose oxidase prepared in serum-free minimal essential medium (MEM). For these studies, endothelial surfaces of isolated bovine corneas were incubated with glucose (1 mg/ml) and glucose oxidase (20 U/ml) for 4, 6, and 10 hours at 37 C/in a 5% CO2 atmosphere. Supernatants were then removed and assayed for bovine neutrophil or mononuclear cell chemotactic activity. C5 fragment was our positive control for 100% chemotactic response. Corneas were also fixed in buffered formalin for histopathologic evaluation. Results of these studies indicated that 1) 6-hour interactions of the glucose (G)/glucose oxidase (GO) mixture with endothelial surfaces resulted in both endothelial cell injury (cytoplasmic vacuolization and convoluted nuclei) and production of chemotactic factors (via checkerboard analysis) specific for both neutrophils (58% maximum chemotactic response [MCR]) and mononuclear cells (75% MCR); 2) control corneas treated with either G or GO for 4 and 6 hours produced low levels of LCFs (5-15% MCR); 3) preliminary molecular weight characterization of cornea-derived LCFs obtained from corneas incubated with G/GO for 6 hours revealed the detection of chemotactic activity specific for mononuclear cells in two major fractions, one near the void volume (greater than 130,000 daltons) and one near the elution volume (less than or equal to 10,000-15,000 daltons). Chemotactic activity specific for neutrophils was detected only in one major fraction near the elution volume (less than or equal to 10,000-15,000 daltons); and 4) the production of these LCFs by isolated corneas was significantly inhibited, in a dose-response fashion, when the enzyme catalase (1200-6000 U/ml) was added to corneas incubated with G/GO for 6 hours. To investigate whether isolated CECs were capable of producing LCFs in response to G/GO injury, the authors incubated cultured bovine CECs with G/GO for 3, 6, and 20 hours at 37 C in a 5% CO2 atmosphere. Similar to isolated corneas, cultured CECs incubated with G/GO for 6 hours produced significant levels of LCFs specific for neutrophils (67% MCR) and mononuclear cells (75% MCR). Furthermore, the addition of catalase (3000 U/ml) to corneas incubated with G/GO for 6 hours markedly reduced the production of LCFs. These in vitro studies suggest that corneal cells and/or corneal matrix may play important roles in the initiation and amplification of endocular inflammation in vivo by elaborating factors that control leukocyte influx to the anterior chamber region.

The purpose of this study was to compare the low-cycle fatigue (LCF) behavior of electropolished and nonelectropolished nickel-titanium (NiTi) instruments of the same design in hypochlorite. Forty-five electropolished and 62 nonelectropolished NiTi engine files were subjected to rotational bending at various curvatures in 1.2% hypochlorite solution. Number of revolutions to failure, crack-initiation sites, extent of slow crack extension into the fracture cross-section, and surface-strain amplitude were noted. A linear relationship was found between LCF life and surface-strain amplitude for both groups, with no discernible difference between the two (p>> 0.05). No electropolished instrument showed more than one crack origin, significantly fewer than for the nonelectropolished instruments (p < 0.05). The square root of crack extension and strain amplitude were inversely related. Although surface smoothness is enhanced by electropolishing, this did not protect the instrument from LCF failure.

It is known that ischemia commonly increases exogenous glucose utilization by accelerating glucose uptake and flux rates through the Embden-Meyerhof pathway. Constitutive enzymes regulate the rate of glycolysis and in turn are regulated by product inhibition and allosteric controls. The purpose of this report was to test whether mRNA abundance for select glycolytic enzymes, and glucose transport proteins, is also modified. Six intact working pig hearts with coronary flow controlled by extracorporeal perfusion were compared at the following conditions: (1) aerobic control perfusion; (2) ischemia affected by a 60% decrease in left anterior descending (LAD) coronary perfusion: (3) ischemia again affected by a 60% decrease in LAD flow followed by a 40-min interval of aerobic reflow; (4) an intermittent ischemia and reflow protocol including four cycles of similar LAD flow reductions (5 min per cycle) interspersed with 15-20 min of aerobic reperfusion; (5) a 4-day model designed to produce myocardial chronic hibernation: and (6) mild ischemia induced by a 40% decrease in LAD flow for 85 min to produce certain adaptations compatible with short-term hibernation. In each heart, mRNA abundance was measured from LAD and circumflex (LCF) perfused myocardium for hexokinase, phosphofructokinase, glyceraldehyde-3-phosphate dehydrogenase and the two glucose transporter isomers, GLUT 4 and GLUT 1. mRNA data from LAD myocardium in intervention hearts were normalized to those from LAD tissue in the control heart (LADc) and with LCF values in the same intervention hearts. Signal variance around unity in the LAD tissue, with respect to that of the LCF myocardium, in the control heart compared closely (44 and 41% in two separate runs, respectively). GLUT 1/GLUT 4 ratios in the LAD and LCF beds of this heart also agreed closely. LAD/LADc ratios were increased for hexokinase (1.69), phosphofructokinase (3.69), and glyceraldehyde-3-phosphate dehydrogenase (2.29) in the ischemia heart and for phosphofructokinase (3.90), glyceraldehyde-3-phosphate dehydrogenase (2.20), GLUT 4 (1.55) and GLUT 1 (2.20) in the ischemia/reflow heart. There was no evidence of excess signal in the intermittent ischemia/reflow, chronic hibernation, or mild ischemia hearts. Altered signal from LCF myocardium was also suggested. These data indicate that mRNA abundance for select glycolytic enzymes and transporter proteins is increased in ischemic myocardium with or without reperfusion and offers a possible mechanism for increased protein activity in settings of diminished regional coronary flow.

A major task in computational analysis of mRNA expression profiles is definition of relationships among profiles on the basis of similarities among them. This is generally achieved by pattern recognition in the distribution of data points representing each profile in a high-dimensional space. Some drawbacks of commonly used pattern recognition algorithms stem from their use of a globally linear space and/or limited degrees of freedom. A pattern recognition method called Local Context Finder (LCF) is described here. LCF uses nonlinear dimensionality reduction for pattern recognition. Then it builds a network of profiles based on the nonlinear dimensionality reduction results. LCF was used to analyze mRNA expression profiles of the plant host Arabidopsis interacting with the bacterial pathogen Pseudomonas syringae. In one case, LCF revealed two dimensions essential to explain the effects of the NahG transgene and the ndr1 mutation on resistant and susceptible responses. In another case, plant mutants deficient in responses to pathogen infection were classified on the basis of LCF analysis of their profiles. The classification by LCF was consistent with the results of biological characterization of the mutants. Thus, LCF is a powerful method for extracting information from expression profile data.

BACKGROUND

Cholesteatoma is traditionally diagnosed by otoscopic examination and treated by surgery. The necessity for imaging in an uncomplicated case is controversial. This study was planned to investigate the usefulness of a preoperative high-resolution computed tomography (HRCT) scan in depicting the status of middle ear structures in the presence of cholesteatoma and also to compare the correspondence between pre- and intraoperative CT findings in patients with cholesteatoma.

METHODS

This prospective descriptive study was performed from January 2009 to May 2011 in 36 patients with cholesteatoma who were referred to the Kashani and Al-Zahra Clinics of Otolaryngology. Preoperative high-resolution temporal bone CT scans (axial and coronal views) were carried out and compared with intraoperative findings.

RESULTS

Evaluation of 36 patients and their CT scans revealed excellent correlation for sigmoid plate erosion, widening of aditus, and erosion of scutum; good correlation for erosion of malleus and tegmen; moderate correlation for lateral canal fistula (LCF) and erosion of mastoid air cells; and poor correlation for facial nerve dehiscence (FND), incus, and stapes erosion.

CONCLUSIONS

A preoperative CT scan may be helpful in relation to diagnosis and decision making for surgery in cases of cholesteatoma and ossicular erosion. The CT scan can accurately predict the extent of disease and is helpful for detection of lateral canal fistula, erosions of dural plate, and ossicular erosions. However it is not able to distinguish between cholesteatoma and mucosal disease, facial nerve dehiscency, incus, and stapes erosion.

This study examined the low-cycle fatigue (LCF) behavior of a nickel-titanium (NiTi) engine-file under various environmental conditions. One brand of NiTi instrument was subjected to rotational-bending fatigue in air, deionized water, sodium hypochlorite, or silicone oil. The curvature of each instrument, diameter of the fracture cross-section, and the number of rotations to failure were determined. The strain-life relationship in the LCF region was examined by using one-way analysis of variance, and the number of crack origins with chi2, for differences between groups. The results showed a linear relationship, on logarithmic scales, between the LCF life and the surface strain amplitude; regression line slopes were significantly different between noncorrosive (air, silicone oil) and corrosive (water, hypochlorite) environments (P < .05), as well as number of crack origins (P < .05). Hypochlorite was more detrimental to fatigue life than water. In conclusion, environmental conditions significantly affect the LCF behavior of NiTi rotary instruments. Fatigue testing of NiTi engine-files should be in a service-like environment.

From 1977-1982, 161 patients were treated using hyperthermia as an adjuvant in Phase I trials. Microwave applicators (MW), capacitively coupled plates (RF plates), interstitial localized current fields (LCF), and magnetic induction heating (MI) techniques were used together with radiation in 135 patients, with chemotherapy in 10 patients, and alone in 16 patients. Tumor volume response categories were no response (NR, less than 50% decrease); partial response (PR, 50% less than or equal to volume decrease less than 100%); and complete response (CR, complete disappearance). The CR rates and total response rates (CR + PR) were 38/160 (24%) and 90/160 (56%), respectively. There were highly significant differences among techniques in CR vs PR + NR (p = .001), and in CR + PR vs NR (p less than .0005). Response did not vary significantly with histologic category. Overall toxicity was 16%, and did not vary significantly with technique (p = .193). In the patient group treated with hyperthermia and radiation, multivariate analysis revealed that a set of three variables had prognostic importance for CR: technique (p = .011), radiation dose (p = .019), and tumor volume (p = .001, negatively correlated). A good correlation also existed between CR and the minimum tumor temperature averaged over all treatments, TMIN (p less than .0005). Temperature variables themselves were correlated with tumor volume. Minimum T correlated negatively with volume (p = .017) and TMAX correlated positively with volume (p = .026). In fewer than 50% of patients could minimum T greater than 40.7 degrees C be achieved. Our conclusions are: TMIN, tumor volume, radiation dose, and heating technique have prognostic value for initial response; variation in CR vs technique reflects variation in tumor volume treated and in minimum temperature achieved with these techniques; and acute toxicity of treatment is infrequent, but serious toxicity is possible with the interstitial technique.

The aim of this study was to investigate the potential anti-cancer effects of probiotic cell-free supernatant (CFS) treatment using Lactobacillusfermentum for colorectal cancer (CRC) in 3D culture systems. Cell viability was assessed using MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assays, whereas apoptosis was monitored through RT-qPCR analysis of Bax, Bak, Noxa, and Bid mRNA expressions in addition to flow cytometry analysis of Lactobacillus cell-free supernatant (LCFS) treatment. Our results showed that the anti-cancer effect of LCFS on cell viability was pronouncedly enhanced in 3D-cultured HCT-116 cells, which was linked to the increased level of cleaved caspase 3. Additionally, upregulation of apoptotic marker gene mRNA transcription was dramatically increased in 3D cultured cells compared to 2D systems. In conclusion, this study suggests that LCFS enhances the activation of intrinsic apoptosis in HCT-116 cells and the potential anti-cancer effects of Lactobacilli mixtures in 3D culture systems. All in all, our study highlights the benefits of 3D culture models over 2D culture modeling in studying the anti-cancer effects of probiotics.

The opioid peptides beta-endorphin and met-enkephalin have been shown to modulate human lymphocyte proliferation, mononuclear cell locomotion, natural killer cell activity, and neutrophil locomotion. This study demonstrates that beta-endorphin and met-enkephalin inhibit the production of a T lymphocyte chemotactic factor (LCF) by concanavalin A (Con A)-stimulated peripheral blood mononuclear cells. Inhibition of LCF production was observed by using concentrations of 10(-11) to 10(-6) M beta-endorphin or met-enkephalin but not alpha-endorphin. A bimodal pattern of suppression of LCF production was observed with both met-enkephalin and beta-endorphin when titrated from 10(-12) to 10(-6) M concentrations, with the peaks of suppressive activity occurring at concentrations of 10(-11) M and 10(-6) M. Timed studies of the production of LCF over a 54-hr period showed that there was an appreciable lag in the onset of measurable LCF activity in mononuclear supernatants produced in the presence of beta-endorphin and met-enkephalin. The suppression of LCF production mediated by opioid peptides in mononuclear supernatants was abrogated by depletion of glass-adherent mononuclear cells before culturing with opioids and Con A. The inhibitory effect of opioid peptides on LCF production was prevented by the addition of indomethacin to cell cultures. Additional experiments showed that exogenous prostaglandin E2 (PGE2) suppressed Con A-stimulated LCF production when added at concentrations ranging from 10(-6) to 10(-8) M. Other studies suggested that the mechanism of opioid peptide-mediated suppression of LCF production was due to an enhanced sensitivity of mononuclear cells to the inhibitory action of PGE2. These data provide further evidence for modulation of the immune response in humans by the neuroendocrine hormones beta-endorphin and met-enkephalin and further suggest a link between this modulation and arachidonic acid metabolism.

Retrospective analysis of data from 14,528 lung cancer patients with multiple primary malignant neoplasm (MPMN) revealed that 2.5% (364/14,528) were MPMN cases and 96.2% (350/364) were diagnosed with two primary malignancies, 3.6% (13/364) with three primary malignancies, and 0.3% (1/364) with four primary malignancies. Among 350 lung cancer patients diagnosed with two primary malignancies, 26.6% (93/350) had lung cancer diagnosed first (LCF) and 73.4% (257/350) had other cancers diagnosed initially (OCF), whereas synchronous MPMN (SMPMN) accounted for 21.1% (74/350) and metachronous MPMN (MMPMN) accounted for 78.9% (276/350) of the cases. Detection of first primary neoplasms were at an early stage for LCF patients and the age of the first lung cancer diagnosis was 59.3 years vs. 55.4 years in the OCF group (P = 0.008), whereas the onset age of second primary neoplasm diagnosis was similar in both groups (62.5 and 61.6 years, P = 0.544). Median survival times of MMPMN and SMPMN patients in the LCF group were 6.83 and 2.42 years and in the OCF group 8.67 years and 2.25 years, respectively. Multivariate analysis showed that SMPMN, LCF and the age of the primary cancer diagnosed first ( ≥ 60 years) and NSCL staging > II were significant independent factors for inferior prognosis of patients.