[Prognosis in elderly patients with falls treated in emergency departments: The EDEN-3 study]. / Pronóstico en pacientes ancianos con caídas atendidos en servicios de urgencias: estudio EDEN-3.

OBJECTIVE: To investigate whether falls in people ≥65 years old are a prognostic factor for adverse events compared to the rest of older patients who consult emergency departments, and identify factors related to a worse long-term evolution. METHOD: EDEN cohort that included patients ≥65 years old. Those patients who consulted for fall and the rest were distinguished. Twelve variables were collected. For comparison: two groups matched by fall propensity score. We compared mortality at one year and combined adverse event post-discharge at one year. In patients with falls, variables independently related to evolution were identified. RESULTS: Two thousand seven hundred and forty-five patients treated for falls and 22,920 for other reasons. Mortality at one year was 14.4% (9.5% vs. 15.0%, respectively, P<.001) and the combined post-discharge adverse event at one year was 60.6% (52.2% vs. 61.7%, respectively, P<.001). In 4748 patients matched by fall propensity score (2372 in each group), the inverse association between consultation for fall and mortality (HR: 0.705, 95% CI: 0.5880.846) and post-discharge combined adverse event (0.758, 0.701-0.820) remained significant. Factors associated with mortality in patients with falls were ≥80 years (2.097, 1.521-2.891) and comorbidity (2.393, 1.574-3.636) while being female was a protective factor (0.758, 0.584-0.985). Between the factors associated with post-discharge combined adverse hospitalization in the index event was a protective factor (0.804, 0.685-0.943). CONCLUSIONS: Patients over 65 years of age treated in the emergency room for falls have a better prognosis. Hospitalization was a protective factor of combined postdischarge adverse event.

Vitamin d deficiency with high parathyroid hormone levels is related to late onset SEPSIS among preterm infants.

Preterm infants (PTs) are at greater risk for vitamin D deficiency, which relates to the possibility of a higher incidence of comorbidities. Our goal was twofold, 1) to monitor vitamin D, calcium, phosphorus, parathyroid hormone (PTH), matrix metalloproteinase-8 (MMP-8) serum levels at three-time points during hospitalization, and 2) to assess the association between 25-hydroxyvitamin D (25OHD) levels and outcomes in PTs. METHODS: We carried out a follow-up on 50 Caucasian PTs ≤ 32 weeks of gestational age (GA) and/or ≤ 1500 g birth weight at 28 days and at 4 months. PTs were divided into two subgroups for tests of association with clinical outcomes based on vitamin D deficient infants 25(OH) D cord blood levels: ≤ 20 ng/ml). At an initial stage, 25(OH) D levels were determined in maternal/preterm blood samples and were compared to full term delivery infants. RESULTS: There were no differences in 25(OH) D serum levels at birth when comparing PTs to term infants, or regarding maternal levels. A strong positive correlation was detected between maternal and neonatal 25(OH) D serum levels among PTs and term infants (r: 0.466; p < 0.001). Neonates with vitamin D deficiency did not present a higher incidence of comorbidities. PTs were classified in two subgroups based on vitamin D and PTH (group 1: vitamin D < 20 ng/mL and PTH > 60 pg/mL; group 2: vitamin D > 20 and PTH < 60 pg/mL). The PTs in group 1 showed a higher incidence of LOS (RR: 2; 95% CI: 1.31-3.55). No relationship was observed between MMP-8 serum levels and the incidence of sepsis. CONCLUSIONS: This study did not find any evidence of an increase in preterm birth risk related to vitamin D level at birth. Vitamin D deficiency by itself is not associated with a higher incidence of comorbidities. However, the binomial vitamin D-PTH must be taken into consideration.

Hormone replacement therapy in children with growth hormone deficiency: impact on immune profile.

Growth hormone (GH) may influence the immune system. The aim of this study was to assess the immune profile after treatment with GH in pre-pubertal children with a deficiency of this hormone. The study was carried out in two phases. Two groups were included in the first phase: group A) children treated with GH; group B) untreated children, prior to starting treatment. In the second phase, group B children were assessed 6 months after starting treatment. In the first phase, groups A and B were compared (case-control study). In the second phase, group B was compared in terms of baseline and final times (before and after study). We analysed: humoral immunity (immunoglobulin (Ig)A, IgG, IgM, C1 inhibitor, C3, and C4) and cell-mediated immunity (CD3+, CD4+, CD8+, CD4+/CD8+, CD19+ lymphocytes, and natural killer (NK) cells). Insulin-like growth factor 1 (IGF-1) and insulin-like growth factor-binding protein 3 (IGFBP-3) were also determined. In the first phase, CD3+, CD4+, CD19+, and NK cells and CD4+/CD8+ were greater in the treated group. CD8+ was lower in this group. No variations were seen in immunoglobulins and the complement between both groups. There were no changes to the complement in either of the two phases. In the second phase, untreated patients were assessed after 6 months of treatment. When comparing the baseline and final immune profiles, a statistically significant decrease in IgG and IgM was observed, and an increase of IGF-1 levels and monocytes. In conclusion, our study shows changes in the cellular and humoral immune profiles in children with GH deficiency who were treated.

Common Variants in 22 Genes Regulate Response to Metformin Intervention in Children with Obesity: A Pharmacogenetic Study of a Randomized Controlled Trial.

Metformin is a first-line oral antidiabetic agent that has shown additional effects in treating obesity and metabolic syndrome. Inter-individual variability in metformin response could be partially explained by the genetic component. Here, we aimed to test whether common genetic variants can predict the response to metformin intervention in obese children. The study was a multicenter and double-blind randomized controlled trial that was stratified according to sex and pubertal status in 160 children with obesity. Children were randomly assigned to receive either metformin (1g/d) or placebo for six months after meeting the defined inclusion criteria. We conducted a post hoc genotyping study in 124 individuals (59 placebo, 65 treated) comprising finally 231 genetic variants in candidate genes. We provide evidence for 28 common variants as promising pharmacogenetics regulators of metformin response in terms of a wide range of anthropometric and biochemical outcomes, including body mass index (BMI) Z-score, and glucose, lipid, and inflammatory traits. Although no association remained statistically significant after multiple-test correction, our findings support previously reported variants in metformin transporters or targets as well as identify novel and promising loci, such as the ADYC3 and the BDNF genes, with plausible biological relation to the metformin's action mechanism. Trial Registration: Registered on the European Clinical Trials Database (EudraCT, ID: 2010-023061-21) on 14 November 2011 (URL: https://www.clinicaltrialsregister.eu/ctr-search/trial/2010-023061-21/ES).

Effectiveness and safety of olive oil preparation for topical use in pressure ulcer prevention: Multicentre, controlled, randomised, and double-blinded clinical trial.

This non-inferiority, multicentre, randomised, controlled, and double-blinded clinical trial compared the therapeutic effectiveness of the topical application of an olive oil solution with that of a hyperoxygenated fatty acid compound for the prevention of pressure ulcers in at-risk nursing home residents. The study population comprised 571 residents of 23 nursing homes with pressure ulcer risk, randomly assigned to a hyperoxygenated fatty acid group (n = 288) or olive oil solution group (n = 283). Both solutions were applied on at-risk skin areas every 12 hours for 30 days or until pressure ulcer onset. The main outcome variable was the pressure ulcer incidence. The absolute risk difference was estimated (with 95% CI) using Kaplan-Meier survival and Cox regression curves. The groups did not significantly differ in any study variable at baseline. The pressure ulcer incidence was 4.18% in the olive oil group vs 6.57% in the control group, with an incidence difference of -2.39% (95% CI = -6.40 to 1.56%), which is within the pre-established non-inferiority margin of ±7%, thus supporting the study hypothesis. We present the first evidence of the effectiveness and safety of the topical application of olive oil to prevent pressure ulcers in the institutionalised elderly.

Electrochemical Synthesis and Magnetic Properties of MFe2O4 (M = Fe, Mn, Co, Ni) Nanoparticles for Potential Biomedical Applications.

In this study, we evaluate the magnetic properties and cytotoxic effect of magnetic nanoparticles (MNPs) based on magnetite and Mn, Co and Ni ferrites, obtained by electrochemical synthesis. These nanoparticles have almost spherical shape and an mode size of 9±1 nm. The electrochemical synthesis produces a single crystallographic phase with a spinel-like structure in all cases. Magnetization saturation at room temperature varies with the composition of the ferrites from MS (Fe3O4) > MS (MnFe2O4) > MS (CoFe2O4) > MS (NiFe2O4). Ferrite MNPs present low magnetic remanence indicating a superparamagnetic-like response at room temperature. However, the different values of magnetic anisotropy and size produce variations in the values of coercivity and susceptibility of the ferrite MNPs. The cytotoxicity of the different ferrites was evaluated by internalizing MNP in HeLa cancer cells. Although magnetite and Mn ferrite present low toxicity for all the concentrations studied, significant cytotoxic effect were observed when incubating the cells with high concentration of Co and Ni ferrites.

Short-Term Evaluation of Left Ventricular Mass and Function in Children With Growth Hormone Deficiency After Replacement Treatment.

Background: Our study was designed to assess the effects of GHD on nutritional and metabolic parameters, brain natriuretic peptide (BNP) levels, and left ventricular mass (LVM) in prepubertal children and after short-term GH replacement therapy. Materials and Methods: This prospective study enrolled 81 children. We compared 40 GHD children (16 males and 24 females) to 41 healthy children (control group) (18 males and 23 females). All subjects were at Tanner Stage I (aged 7-11 years). At the baseline, a blood sample was drawn and echocardiographic images were obtained. These tests were repeated on the GHD subjects after 6 months of GH replacement therapy. Body surface, weight, size, blood pressure, heart rate, glucose, insulin, HOMA-IR, HOMA-ß, QUICKI, cholesterol, HDLc, LDLc, triglycerides, IGF1, and IGFBP3 were measured. Indexed LVM, diastolic and systolic diameter (dD-sD), diastolic and systolic LV function, isovolumic relaxation time, right ventricle function, and BNP levels were obtained through echocardiography. These parameters were correlated to growth factors. Data were analyzed using Student's t-test or U-Mann-Whitney-test and Pearson's correlation, considering p < 0.05 to be significant. Results: Indexed LVM was smaller in GHD patients than in controls, whereas diastolic and systolic functions, BNP, metabolic, and nutritional profiles were similar. After treatment, nutritional and metabolic profiles significantly improved, though diastolic and systolic functions did not seem to have changed. There was a significant increase in LVM. Indexed LVM was similar to that of controls. Significant correlations were obtained between LVM-IGF1 and sD-IGFBP3. Conclusions: GHD in childhood is associated with a lower indexed LVM. In the short-term, GH increases the indexed LVM, while maintaining normal systolic and diastolic functions, BNP, and an improved lipid profile.

Metformin for Obesity in Prepubertal and Pubertal Children: A Randomized Controlled Trial.

OBJECTIVES: Metformin has shown its effectiveness in treating obesity in adults. However, little research has been conducted in children, with a lack of attention on pubertal status. The objectives were to determine whether oral metformin treatment reduces BMI z score, cardiovascular risk, and inflammation biomarkers in children who are obese depending on pubertal stage and sex. METHODS: This was a randomized, prospective, double-blind, placebo-controlled, multicenter trial, stratified according to pubertal stage and sex, conducted at 4 Spanish clinical hospitals. Eighty prepubertal and 80 pubertal nondiabetic children who were obese aged 7 to 14 years with a BMI >95th percentiles were recruited. The intervention included 1 g/d of metformin versus placebo for 6 months. The primary outcome was a reduction in BMI z score. Secondary outcomes comprised insulin resistance, cardiovascular risk, and inflammation biomarkers. RESULTS: A total of 140 children completed the study (72 boys). Metformin decreased the BMI z score versus placebo in the prepubertal group (-0.8 and -0.6, respectively; difference, 0.2; P = .04). Significant increments were observed in prepubertal children treated with metformin versus placebo recipients in the quantitative insulin sensitivity check index (0.010 and -0.007; difference, 0.017; P = .01) and the adiponectin-leptin ratio (0.96 and 0.15; difference, 0.81; P = .01) and declines in interferon-γ (-5.6 and 0; difference, 5.6; P = .02) and total plasminogen activator inhibitor-1 (-1.7 and 2.4; difference, 4.1; P = .04). No serious adverse effects were reported. CONCLUSIONS: "Metformin decreased the BMI z score and improved inflammatory and cardiovascular-related obesity parameters only in prepubertal children, but a differential effect of metformin was not observed in prepubertal compared to pubertal children. Nevertheless, the doses per kilogram of weight administrated may have had an impact on the metformin effect. Further investigations are necessary."

Morbility, clinical data and proteomic analysis of IUGR and AGA newborns at different gestational ages.

The data are related to the proteomic analysis of 43 newborns with intrauterine growth retardation (IUGR) and 45 newborns with appropriate weight for gestational age (AGA) carried out by separation via 2DE and analyzed by MS-TOF/TOF. All newborns were separated into three gestational age groups, "Very Preterm" 29-32 weeks, "Moderate Preterm" 33-36 weeks, and, "Term" ≥37weeks. From each newborn, blood was drawn three times from birth to 1 month life. High-abundant serum proteins were depleted, and the minority ones were separated by 2DE and analyzed for significant expression differences. The data reflect analytic and clinic variables analyzed globally and categorized by gestational age in relation to IUGR and the optimization of conditions for 2-DE separation. The data from this study are related to the research article entitled "Alterations of Protein Expression in Serum of Infants with Intrauterine Growth Restriction and Different Gestational Ages" (M.D. Ruis-González, M.D. Cañete, J.L. Gómez-Chaparro, N. Abril, R. Cañete, J. López-Barea, 2015) [1]. The present dataset of serum IUGR newborn proteome can be used as a reference for any study involving intrauterine growth restriction during the first month of life.

Evaluation of differential effects of metformin treatment in obese children according to pubertal stage and genetic variations: study protocol for a randomized controlled trial.

BACKGROUND: Overweight and obesity are considered to be serious public health problems. In pediatric populations, insulin resistance, dyslipidemia, and hypertension associated with obesity occur with increased frequencies. Metformin is an oral anti-hyperglycemic agent that has been demonstrated to be efficacious in the treatment of diabetic and non-diabetic obese adults. A considerable amount of pharmacogenetic research has demonstrated that genetic variation is one of the major factors affecting metformin response. Additionally, potential microbiota-mediated mechanisms of metformin effect have been recently described. However, scant work has been conducted in children, with no attention being paid to the potential effects of pubertal development. Thus, the main objective of the present study is to evaluate the effect of metformin treatment together with lifestyle recommendations in a randomized control trial (RCT) of obese children according to pubertal stage, genetic variants and signature of gut microbiota. METHODS/DESIGN: This is a randomized, prospective, double-blind, placebo-controlled, multicenter trial, which is stratified by puberty and sex. Eighty pre-pubertal (40 boys and 40 girls) and 80 pubertal non-diabetic obese children (40 boys and 40 girls) are being recruited in four Spanish Clinical Hospitals. The inclusion criteria to participate in the RCT include a Body Mass Index (BMI) above the 95th percentile and age 7-14 years. The pubertal stage is determined based on the Tanner criteria. Participants are assigned to two groups in accordance with a randomization schedule and receive 1 g of metformin or placebo for six months in combination with healthy lifestyle recommendations in both groups. The primary outcomes include changes in the BMI Z score and the biomarkers associated with the early appearance of insulin resistance syndrome, inflammation, cardiovascular risk according of the presence of genetic determinants of metformin response, as well as possible modifications in microbiota. DISCUSSION: This study will assess the differential response of metformin treatment at six months in pre-pubertal and pubertal obese children. TRIAL REGISTRATION: Registered by European Clinical Trials Database (EudraCT, ID: 2010-023061-21) on 14 November 2011.

Two combined photosensitizers: a goal for more effective photodynamic therapy of cancer.

Photodynamic therapy (PDT) is a clinically approved therapeutic modality for the treatment of diseases characterized by uncontrolled cell proliferation, mainly cancer. It involves the selective uptake of a photosensitizer (PS) by neoplastic tissue, which is able to produce reactive oxygen species upon irradiation with light, leading to tumor regression. Here a synergistic cell photoinactivation is reported based on the simultaneous administration of two PSs, zinc(II)-phthalocyanine (ZnPc) and the cationic porphyrin meso-tetrakis(4-N-methylpyridyl)porphine (TMPyP) in three cell lines (HeLa, HaCaT and MCF-7), using very low doses of PDT. We detected changes from predominant apoptosis (without cell detachment) to predominant necrosis, depending on the light dose used (2.4 and 3.6 J/cm(2), respectively). Analysis of changes in cytoskeleton components (microtubules and F-actin), FAK protein, as well as time-lapse video microscopy evidenced that HeLa cells were induced to undergo apoptosis, without losing adhesion to the substrate. Moreover, 24 h after intravenous injection into tumor-bearing mice, ZnPc and TMPyP were preferentially accumulated in the tumor area. PDT with combined treatment produced significant retardation of tumor growth. We believe that this combined and highly efficient strategy (two PSs) may provide synergistic curative rates regarding conventional photodynamic treatments (with one PS alone).

[Arterial pre-hypertension in young adults]. / Prehipertensión arterial en adultos jóvenes.

BACKGROUND AND OBJECTIVES: Prehypertension is a new category of blood pressure and is considered a cardiovascular risk factor. This study has aimed to estimate the prevalence of prehypertension and the association between prehypertension and other vascular risk factors in young adults. MATERIAL AND METHODS: First year university students from all areas of study in the University of Cuenca were invited to participate. Prehypertension was defined as systolic blood pressure between 120-139 mmHg and/or diastolic blood pressure between 80-89 mmHg. Anthropometric, lipid and metabolic variables were measures. The presence of metabolic syndrome was evaluated and quantified based on the sum of the standardized scores of the waist circumference, the triglyceride/c-HDL ratio, mean blood pressure and R-HOMA (Index of insulin resistance to glucose lowering effect). RESULTS: A total of 545 university students were included in the analysis (mean age 20.36±3.9 years, 74.7% women). Prehypertension prevalence was 24% (95% CI: 21-27%), (56.5% in men and 13% in women). The condition of prehypertension was directly associated to the body mass index (OR: 1.194; 95% CI: 1.124-1.311), insulin resistance (R-HOMA, OR: 2.638; 95% CI: 1.263-4.926) and to the index or quantification of the severity of the metabolic syndrome (OR: 4-868; 95% CI: 3-846-8-328). On the other hand, HDL-c showed an inverse relationship with prehypertension (OR: 0.981; 95% CI: 0.957-0.993). CONCLUSIONS: One out of every four young adults presents prehypertension. This condition is associated to well-established vascular risk factors.

An artificial neural network model for predicting the subcellular localization of photosensitisers for photodynamic therapy of solid tumours.

Photodynamic therapy (PDT) is a promising modality for the treatment of tumours based on the combined action of a photosensitiser (PS), visible light and molecular oxygen, which generates a local oxidative damage that leads to cell death. The site where the primary photodynamic effect takes place depends on the subcellular localization of the PS and affects the mode of action and efficacy of PDT. It is therefore of prime interest to develop structure-subcellular localization prediction models for a PS from its molecular structure and physicochemical properties. Here we describe such a prediction method for the localization of macrocyclic PSs into cell organelles based on a wide set of physicochemical properties and processed through an artificial neural network (ANN). 128 2D-molecular descriptors related to lipophilicity/hydrophilicity, charge and structural features were calculated, then reduced to 76 by using Pearson's correlation coefficient, and finally to 5 using Guyon and Elisseeff's algorithm. The localization of 61 PSs was compiled from literature and distributed into 3 possible cell structures (mitochondria, lysosomes and "other organelles"). A non-linear ANN algorithm was used to process the information as a decision tree in order to solve PS-organelle assignment: first to identify PSs with mitochondrial and/or lysosomal localization from the rest, and to classify them in a second stage. This sequential ANN classification method has permitted to distinguish PSs located into two of the most important cell targets: lysosomes and mitochondria. The absence of false negatives in this assignation, combined with the rate of success in predicting PS localization in these organelles, permits the use of this ANN method to perform virtual screenings of drug candidates for PDT.

[Usefulness of electrocardiographic screening in a neonatal population]. / Utilidad del cribado electrocardiográfico en población neonatal.

INTRODUCTION: Congenital long QT syndrome is a rare disease, but is responsible for nearly 10% of Sudden Infant Death Syndrome. It is characterized by an abnormal prolonged QT interval in the basal electrocardiogram (ECG) with life-threatening arrhythmias which occur in previously asymptomatic patients and are preventable with an appropriate treatment. AIMS: The impact of introducing ECG-screening in newborns is studied and main ECG-measurements are described in our population. MATERIAL AND METHODS: Twelve-lead ECG was carried out. MEASUREMENTS: RR, PR and QT interval, heart rate corrected QT interval, R wave voltage in V1, AVR and AVL, Q wave in I and AVL, P amplitude and voltage, right bundle branch block and ST elevation (Brugada pattern) and delta wave. It was considered pathological: QTc >0.44 or <0.30 seconds; R >12 in V1 and >8mm in AVR; R >7.5mm in AVL; Q >25% QRS in I and AVL; Brugada pattern; delta wave. RESULTS: A total of 1061 healthy children were born in our hospital between 29 May 2007 and 12 December 2008, of which 50.3% were males. An ECG was performed on 1006 (94.8%). Five ECG were pathological (0.5%): 2 long QT interval, 2 Wolf-Parkinson-White, 1 pathological Q-wave. A second ECG confirmed except for 2 long QT. No structural heart disease was found. CONCLUSIONS: ECG-screening in newborns is an innocuous, low-cost and parent-well-accepted test that allows us to diagnose asymptomatic but potentially lethal and preventable heart disease; main intervals and waves in our population are describes in this study.

Improving images of fluorescent cell labeling by background signal subtraction.

The uptake and selective accumulation of fluorescent labels and drugs into organelles of cultured cells currently are widely investigated in biomedical research. In such studies, co-localization procedures are frequently used to identify the accumulation sites of compounds with biological activity. A drawback with fluorescent labeling is the autofluorescence of some cell organelles, which can hinder the precise assessment of co-localization. We report here labeling of the Golgi apparatus of A-549 cells using the photosensitizer zinc(II)-phthalocyanine (ZnPc) and co-localization with the Golgi probe NBD C6-ceramide. The blue autofluorescence signal of mitochondria can be subtracted easily from the original picture by image processing, after which the co-localization of the isolated red ZnPc signal with the green signal from the Golgi probe is considerably improved.

Porphycenes: facts and prospects in photodynamic therapy of cancer.

The photodynamic process induces cell damage and death by the combined effect of a photosensitizer (PS), visible light, and molecular oxygen, which generate singlet oxygen ((1)O(2)) and other reactive oxygen species that are responsible for cytotoxicity. The most important application of this process with increasing biomedical interest is the photodynamic therapy (PDT) of cancer. In addition to hematoporphyrin-based drugs, 2nd generation PSs with better photochemical properties are now studied using cell cultures, experimental tumors and clinical trials. Porphycene is a structural isomer of porphyrin and constitutes an interesting new class of PS. Porphycene derivatives show higher absorption than porphyrins in the red spectral region (lambda > 600 nm, epsilon > 50000 M-(1)cm(-1)) owing to the lower molecular symmetry. Photophysical and photobiological properties of porphycenes make them excellent candidates as PSs, showing fast uptake and diverse subcellular localizations (mainly membranous organelles). Several tetraalkylporphycenes and the tetraphenyl derivative (TPPo) induce photodamage and cell death in vitro. Photodynamic treatments of cultured tumor cells with TPPo and its palladium(II) complex induce cytoskeletal changes, mitotic blockage, and dose-dependent apoptotic or necrotic cell death. Some pharmacokinetic and phototherapeutic studies on experimental tumors after intravenous or topical application of lipophilic alkyl-substituted porphycene derivatives are known. Taking into account all these features, porphycene PSs should be very useful for PDT of cancer and other biomedical applications.

Photodynamic Therapy of the Murine LM3 Tumor Using Meso-Tetra (4-N,N,N-Trimethylanilinium) Porphine.

Photodynamic therapy (PDT) of cancer is based on the cytotoxicity induced by a photosensitizer in the presence of oxygen and visible light, resulting in cell death and tumor regression. This work describes the response of the murine LM3 tumor to PDT using meso-tetra (4-N,N,N-trimethylanilinium) porphine (TMAP). BALB/c mice with intradermal LM3 tumors were subjected to intravenous injection of TMAP (4 mg/kg) followed 24 h later by blue-red light irradiation (λmax: 419, 457, 650 nm) for 60 min (total dose: 290 J/cm(2)) on depilated and glycerol-covered skin over the tumor of anesthetized animals. Control (drug alone, light alone) and PDT treatments (drug + light) were performed once and repeated 48 h later. No significant differences were found between untreated tumors and tumors only treated with TMAP or light. PDT-treated tumors showed almost total but transitory tumor regression (from 3 mm to less than 1 mm) in 8/9 animals, whereas no regression was found in 1/9. PDT response was heterogeneous and each tumor showed different regression and growth delay. The survival of PDT-treated animals was significantly higher than that of TMAP and light controls, showing a lower number of lung metastasis but increased tumor-draining lymph node metastasis. Repeated treatment and reduction of tissue light scattering by glycerol could be useful approaches in studies on PDT of cancer.