Cellular dynamics as a marker of normal-to-cancer transition in human cells.

Normal-to-cancer (NTC) transition is known to be closely associated to cell´s biomechanical properties which are dependent on the dynamics of the intracellular medium. This study probes different human cancer cells (breast, prostate and lung), concomitantly to their healthy counterparts, aiming at characterising the dynamical profile of water in distinct cellular locations, for each type of cell, and how it changes between normal and cancer states. An increased plasticity of the cytomatrix is observed upon normal-to-malignant transformation, the lung carcinoma cells displaying the highest flexibility followed by prostate and breast cancers. Also, lung cells show a distinct behaviour relative to breast and prostate, with a higher influence from hydration water motions and localised fast rotations upon NTC transformation. Quasielastic neutron scattering techniques allowed to accurately distinguish the different dynamical processes taking place within these highly heterogeneous cellular systems. The results thus obtained suggest that intracellular water dynamics may be regarded as a specific reporter of the cellular conditions-either healthy or malignant.

Anthropogenic debris ingestion in a tropical seabird community: Insights from taxonomy and foraging distribution.

Oceans have been considered as an unlimited supply of goods and services, but resource extraction and waste disposal became ubiquitous and have been damaging the health of marine ecosystems. Finding suitable sentinel species of the human impacts on the oceans is thus imperative, since they may work as early warnings of disruptive situations. In this study, we investigated how taxonomy and foraging distribution influenced the occurrence of anthropogenic debris among five seabird species inhabiting the tropical Atlantic region. Occurrence of anthropogenic debris was assessed using faeces of breeding individuals as a proxy of ingestion. A total of 268 particles were extracted from all samples. The categories "fragments" and "fibres", as well as the colour "blue", were the most prevalent characteristics across species. There was a high diversity of polymers from cellulosic particles to synthetic plastics (Anthropogenic Cellulosic 26.9 %; Polyester 7.7 %; Varnish 5.8 %; Polypropylene 1.9 %). Species with a more coastal foraging strategy exhibited higher occurrence and number of anthropogenic debris when compared to species foraging comparably more in pelagic areas. This suggests that anthropogenic debris are more prevalent in coastal foraging areas, where human activities occur in higher number and frequency (e.g., fisheries) and sources of freshwater input from inland are at close distance. These results provide more evidence to the growing perception on the ubiquity and diversity of anthropogenic debris in the marine environment, and further support the usefulness of using seabirds as bio-indicators of anthropogenic pollution in both neritic and oceanic regions.

Water dynamics in human cancer and non-cancer tissues.

Normal-to-malignant transformation is a poorly understood process associated with cellular biomechanical properties. These are strongly dependent on the dynamical behaviour of water, known to play a fundamental role in normal cellular activity and in the maintenance of the three-dimensional architecture of the tissue and the functional state of biopolymers. In this study, quasi-elastic neutron scattering was used to probe the dynamical behaviour of water in human cancer specimens and their respective surrounding normal tissue from breast and tongue, as an innovative approach for identifying particular features of malignancy. This methodology has been successfully used by the authors in human cells and was the first study of human tissues by neutron scattering techniques. A larger flexibility was observed for breast versus tongue tissues. Additionally, different dynamics were found for malignant and non-malignant specimens, depending on the tissue: higher plasticity for breast invasive cancer versus the normal, and an opposite effect for tongue. The data were interpreted in the light of two different water populations within the samples: one displaying bulk-like dynamics (extracellular and intracellular/cytoplasmic) and another with constrained flexibility (extracellular/interstitial and intracellular/hydration layers).

Micellar effects and analytical applications of nitro substitution in 4-Nitro-N-alkyl-1,8-naphthalimide by cysteine derivatives.

The aromatic nucleophilic substitution reactions of the nitro group of 4-Nitro-N-alkyl-1,8-naphthalimides by thiolate anions produce fluorescent derivatives and their rates are strongly accelerated by micelles of hexadecyltrimethylammonium chloride even at low pH. Acceleration factors of this reactions can reach million-fold. As the products are oxidant-insensible, this reaction allows the determination of SH- containing compounds such as cysteine, glutathione or proteins even in oxidative conditions. Limits of detection are as low as 5 × 10-7 M, ten times lower than the limit for the classic 5,5'-dithiobis-(2-nitrobenzoic) acid method. Moreover, this reaction can be developed at pHs between 6.5 and 7.5 thereby diminishing the rate of spontaneous oxidation of the thiols. In addition, we demonstrated that 4-Nitro-N-alkyl-1,8-naphthalimides can be used to evidence SH groups in peptides, proteins and living cells.

Role of intracellular water in the normal-to-cancer transition in human cells-insights from quasi-elastic neutron scattering.

The transition from normal to malignant state in human cells is still a poorly understood process. Changes in the dynamical activity of intracellular water between healthy and cancerous human cells were probed as an innovative approach for unveiling particular features of malignancy and identifying specific reporters of cancer. Androgen-unresponsive prostate and triple-negative breast carcinomas were studied as well as osteosarcoma, using the technique of quasi-elastic neutron scattering. The cancerous cells showed a considerably higher plasticity relative to their healthy counterparts, this being more significant for the mammary adenocarcinoma. Also, the data evidence that the prostate cancer cells display the highest plasticity when compared to triple-negative mammary cancer and osteosarcoma, the latter being remarkably less flexible. Furthermore, the results suggest differences between the flexibility of different types of intracellular water molecules in normal and cancerous cells, as well as the number of molecules involved in the different modes of motion. The dynamics of hydration water molecules remain virtually unaffected when going from healthy to cancer cells, while cytoplasmic water (particularly the rotational motions) undergoes significant changes upon normal-to-cancer transition. The results obtained along this study can potentially help to understand the variations in cellular dynamics underlying carcinogenesis and tumor metastasis, with an emphasis on intracellular water.

Novel platinum-based anticancer drug: a complete vibrational study.

The introduction of cisplatin to oncology, in the 1970s, marked the onset of the search for novel and improved metal-based anticancer drugs. Polynuclear PtII and PdII complexes with linear alkylamines as bridging ligands are a class of potential antineoplastic agents that have shown promising cytotoxicity against low-prognosis human cancers, such as metastatic breast adenocarcinoma and osteosarcoma. The present study reports an analysis of [µ-N,N'-bis(3-aminopropyl)butane-1,4-diamine-κ4N,N':N'',N''']bis[dichloridoplatinum(II)], [Pt2Cl4(C10H26N4)], denoted Pt2Spm (Spm is spermine), by vibrational spectroscopy coupled to theoretical calculations. Within the latter, the Density Functional Theory (DFT - mPW1PW/6-31G*) and Effective Core Potential (ECP - LANL2DZ) approaches were used, in order to ensure the most accurate representation of the molecule and achieve a maximum agreement with the experimental data. The solid-state geometry of Pt2Spm corresponds to Ci symmetry, displaying 132 vibrational modes. A complete assignment of the experimental vibrational profile of the system was attained through the combined application of complementary Raman, FT-IR and Inelastic Neutron Scattering (INS) techniques. INS allowed an unequivocal identification of the CH2 and NH2 rocking modes, not clearly detected by the optical techniques, while Raman measurements led to a clear discrimination of the Pt-N stretching frequencies from the two distinct Pt-N moieties within the chelate. The metal-to-metal distances calculated for the molecule under study were found to allow the establishment of effective inter- and intrastrand crosslinks with DNA. These results will hopefully help to clarify the mode of action of the compound, at the molecular level, contributing to the development of improved cisplatin-like chemotherapeutic drugs having a higher efficacy and specificity coupled to lower acquired resistance and deleterious side effects.

A velocidade de infusão da solução poli-iônica intravenosa contendo 84mEq/L de lactato determina a intensidade do efeito alcalinizante em equinos / The intravenous polyionic solution with 84mEq/L of lactate infusion rate determines the intensity of the alkalizing effect in horses

O objetivo deste trabalho foi verificar os efeitos da solução poli-iônica intravenosa contendo 84mEq/L de lactato (L84) sobre os equilíbrios hidroeletrolítico e ácido-base de equinos, quando administrada de forma rápida ou lenta. Cinco equinos sadios adultos receberam a infusão contínua intravenosa da L84, em volume correspondente a 10% do peso corporal, em duas ocasiões: a) infusão rápida (16,66mL/kg/h) durante seis horas; b) infusão lenta (8,33mL/kg/h) durante 12 horas. Amostras de sangue venoso foram colhidas ao início da infusão (hora zero) e três, seis, nove, 12 e 24 horas após, e amostras de urina nas horas zero, seis, 12 e 24. Determinaram-se pH (sanguíneo e urinário), pCO2, HCO3 -, BE, PPT, lactato L, Na+, K+, Cl-, AG, SID, Atot, VVP, densidade urinária e excreções fracionadas urinárias de lactato L, Na+, K+ e Cl-. A L84 provoca efeito alcalinizante iatrogênico de menor magnitude quando administrada de forma lenta, porque os mecanismos renais, atuantes durante o período de infusão, promovem a correção gradativa do desequilíbrio. Pode-se concluir que a infusão de forma lenta da solução L84 em equinos é recomendável nos casos em que se suspeite de acidose metabólica e não seja possível quantificar o grau do desequilíbrio.(AU)

The aim of this study was to investigate the effects of an intravenous polyionic solution containing 84mEq/L of lactate (L84) on the hydroelectrolyte and acid-base balances when administered quickly or slowly in horses. Five healthy adult horses received the L84 solution, in a volume corresponding to 10% of BW, by continuous intravenous infusion, in two instants: a) rapid infusion (16.66mL/kg/h) during 6 hours; b) slow infusion (8.33mL/kg/h) during 12 hours. Venous blood samples were taken at the beginning of the infusion (hour 0) and 3, 6, 9, 12, and 24 hours after. Urine samples were taken at 0, 6, 12, and 24h. pH (blood and urine), pCO2, HCO3-, BE, TPP, L-lactate, Na+, K+, Cl-, AG, SID, Atot, PVV, urine specific gravity, and L-lactate, Na+, K+, and Cl- renal fractional clearance were determined. The L84 solution causes lower magnitude alkalizing effect when administered slowly, due to the gradual correction of the iatrogenic imbalance by the kidneys during the infusion period. The L84 solution infused at a low rate in horses could be recommended in cases where metabolic acidosis is suspected and it is not possible to quantify the imbalance degree.(AU)

Intracellular water - an overlooked drug target? Cisplatin impact in cancer cells probed by neutrons.

The first neutron scattering study on human nucleated cells is reported, addressing the subject of solvent-slaving to a drug by probing intracellular water upon drug exposure. Inelastic and quasi-elastic neutron scattering spectroscopy with isotope labelling was applied for monitoring interfacial water response to the anticancer drug cisplatin, in the low prognosis human metastatic breast cancer cells MDA-MB-231. Optical vibrational data were also obtained for lyophilised cells. Concentration-dependent dynamical changes evidencing a progressive mobility reduction were unveiled between untreated and cisplatin-exposed samples, concurrent with variations in the native organisation of water molecules within the intracellular medium as a consequence of drug action. The results thus obtained yielded a clear picture of the intracellular water response to cisplatin and constitute the first reported experimental proof of a drug impact on the cytomatrix by neutron techniques. This is an innovative way of tackling a drug's pharmacodynamics, searching for alternative targets of drug action.

The use of papain gel cream and sunflower oil in promoting healing in a wound in dogs: three case reports / Uso de gel creme de papaína na cicatrização de feridas em cães: relato de três casos

Papain is a proteolytic enzyme removed from the leaves of green papaya and/or latex. This enzyme is widely known as a medicinal fruit used extensively in human medicine for the treatment of wounds of various etiologies. However, studies and reports in veterinary medicine are scarce. Another herbal drug widely used in wound healing is Sunflower oil (Helianthus annus). It has inflammatory and antimicrobial properties which stimulate the local neovascularization, promoting tissue granulation, cell migration, fibroblast proliferation, and differentiation. Three dogs were treated with infected necrosis wounds, considered large, extent, and severe, with varied etiology. All cases were treated with papain gel with the exception of one dog, which was given the sunflower oil at the end of the treatment. Papain gel shows effectiveness in the treatment of wounds especially with wound debridement and removal of necrotic tissue. In addition, the healing time was shorter when compared to the treatment with sunflower oil. Finally, the herbal drugs have a low cost and high accessibility. This study contributes to create a new research regarding the use of this drug in animal wound healing.(AU)

A papaína é uma enzima proteolítica retirada do mamão-papaia verde e/ou do látex das folhas do mamoeiro, tendo propriedades medicinais conhecidas na medicina humana para tratamento de feridas de diversas etiologias. Estudos e/ou relatos na medicina veterinária são escassos. Nesse sentido, objetivou-se descrever três casos de cães, que foram atendidos no Hospital Veterinário da Universidade Federal de Lavras, Brasil, todos com feridas infectadas, necrosadas, com etiologia variada e consideradas de grande extensão e gravidade, sendo tratados com gel de papaína. O tratamento demonstrou efetividade na recuperação das feridas, com a formação de grande quantidade de tecido de granulação, em um período menor que os tratamentos convencionais. Por se tratar de um fitoterápico, de baixo custo e alta acessibilidade, o gel de papaína pode ser utilizado no tratamento de feridas de grande extensão em cães e, assim como em humanos, parece ter um efeito benéfico no processo de cicatrização. Pesquisas devem ser conduzidas para elucidar a ação desse produto nos tecidos, bem como as diferentes concentrações a serem administradas.(AU)

Chemotherapeutic response to cisplatin-like drugs in human breast cancer cells probed by vibrational microspectroscopy.

Studies of drug-cell interactions in cancer model systems are essential in the preclinical stage of rational drug design, which relies on a thorough understanding of the mechanisms underlying cytotoxic activity and biological effects, at a molecular level. This study aimed at applying complementary vibrational spectroscopy methods to evaluate the cellular impact of two Pt(ii) and Pd(ii) dinuclear chelates with spermine (Pt2Spm and Pd2Spm), using cisplatin (cis-Pt(NH3)2Cl2) as a reference compound. Their effects on cellular metabolism were monitored in a human triple-negative metastatic breast cancer cell line (MDA-MB-231) by Raman and synchrotron-radiation infrared microspectroscopies, for different drug concentrations (2-8 µM) at 48 h exposure. Multivariate data analysis was applied (unsupervised PCA), unveiling drug- and concentration-dependent effects: apart from discrimination between control and drug-treated cells, a clear separation was obtained for the different agents studied - mononuclear vs. polynuclear, and Pt(ii) vs. Pd(ii). Spectral biomarkers of drug action were identified, as well as the cellular response to the chemotherapeutic insult. The main effect of the tested compounds was found to be on DNA, lipids and proteins, the Pd(ii) agent having a more significant impact on proteins while its Pt(ii) homologue affected the cellular lipid content at lower concentrations, which suggests the occurrence of distinct and unconventional pathways of cytotoxicity for these dinuclear polyamine complexes. Raman and FTIR microspectroscopies were confirmed as powerful non-invasive techniques to obtain unique spectral signatures of the biochemical impact and physiological reaction of cells to anticancer agents.

Low-grade inflammation predicts persistence of depressive symptoms.

RATIONALE: Evidence suggests that depression is cross-sectionally and longitudinally associated with activation of inflammatory response system. A few studies, however, have investigated the longitudinal relationship between raised inflammatory biomarkers and persistence of depressive symptoms. We examined the temporal relationship between serum levels of inflammatory biomarkers and persistence of depressive symptoms among older participants. METHODS: Center for Epidemiologic Studies Depression Scale (CES-D) was used to assess depressive symptoms at baseline and at 5-year follow-up in 656 participants (233 men, 423 women) aged >60 years of the Rotterdam Study. Markers of inflammation interleukin (IL)-6, alpha-1-antichymotrypsin (ACT) and C-reactive protein (CRP) were assessed at baseline, and all participants taking antidepressant medications were excluded from the analysis. RESULTS: No cross-sectional association was found between IL-6, ACT and CRP with depressive symptoms at baseline. However, higher levels of IL-6 and CRP predicted depressive symptoms at 5-year follow-up. Adjustment for confounding variables had no impact on the observed associations. Similarly, a positive association was found between baseline levels of IL-6 (OR = 2.44, p = 0.030) and CRP (OR = 1.81, p = 0.052) and persistence of depressive symptoms over 5 years. CONCLUSION: Our data suggest that dysregulation of the inflammatory response system is associated with a more severe form of depression more likely to re-occur.

Hb40-61a: Novel analogues help expanding the knowledge on chemistry, properties and candidacidal action of this bovine α-hemoglobin-derived peptide.

This study expands the knowledge on chemical synthesis and properties of Hb40-61a as well as provides results of the first steps given towards knowing how it kills Candida cells. For the first time, this peptide, its all-D analogue (D-Hb40-61a) and its fluorescently labeled analogue (FAM-Hb40-61a) were successfully assembled on resin at 60°C using conventional heating in all steps. Purified and characterized, these peptides exhibited very low toxicity on human erythrocytes. Hb40-61a and D-Hb40-61a were equally active against Candida strains, ruling out sterically specific interactions on their working mechanism. Cell permeabilization assays confirmed progressive damage of the yeast plasma membrane with increasing concentrations of Hb40-61a. While experiment using the fluorescent probe DiBAC4(5) revealed that this synthetic hemocidin alters the yeast plasma membrane potential, test employing DPH indicated that Hb40-61a might affect its dynamics. Exposure of the yeast cells to FAM-Hb40-61a showed that the peptide accumulates in the cell membrane at the ½ MIC, but stains about 97% of the cells at the MIC. Such effect is salt-dependent and partially energy-dependent. These new findings indicate that the central target of Hb40-61a in Candida cells is the plasma membrane and that this synthetic hemocidin should be considered as a potential candidacidal for topic uses.

A utilização da radioterapia no tratamento do carcinoma de células escamosas cutâneo felino avançado / Radiation therapy for the treatment of feline advanced cutaneous squamous cell carcinoma

Avaliou-se a eficácia da radioterapia no tratamento de felinos portadores de carcinoma de células escamosas cutâneo avançado. Um protocolo de fracionamento padrão de radioterapia foi aplicado em seis gatos portadores de uma ou mais lesões cutâneas de carcinoma de células escamosas, em um total de sete lesões neoplásicas, confirmadas por meio de análise histológica. Uma lesão foi classificada como T2 e seis como T4, segundo o sistema de estadiamento da Organização Mundial de Saúde para tumores epidermais de felinos. Os animais foram submetidos a doze frações radioterápicas de 4 Gy, realizadas três vezes por semana, utilizando-se um equipamento de ortovoltagem. Utilizaram-se energia de 120 Kv, 15mA e filtro 2mm de alumínio, o cone usado foi de 6x8cm, e a distância foco-pele foi de 30cm. As lesões foram acompanhadas durante todo o tratamento radioterápico e 30 e 60 dias após o seu término. Neste estudo, 87% das lesões resultaram em remissão completa, e 13%, em remissão parcial. Os efeitos colaterais do tratamento radioterápico, de acordo com a tabela de critérios de toxicidade aguda da Veterinary Radiation Therapy Oncology Group, foram leves e reversíveis em todos os casos, e incluíram eritema cutâneo, epilação e rinite. Considerou-se a radioterapia segura para felinos com carcinoma de células escamosas cutâneo, levando a efeitos colaterais leves, e pode representar uma boa opção terapêutica.

The efficacy of radiation therapy for feline advanced cutaneous squamous cell carcinoma was evaluated. A full course radiation therapy protocol was applied to six cats showing single or multiple facial squamous cell carcinomas, in a total of seven histologically confirmed neoplastic lesions. Of the lesions, one was staged as T 1 , and six as T 4 according to WHO staging system of epidermal tumors. The animals were submitted to twelve radiation fractions of 4 Gy each, on a Monday-Wednesday-Friday schedule, and the equipment used was an orthovoltage unit. Energy used was 120 kV, 15 mA and 2 mm aluminum filter. The cats were evaluated duri ng the treatment and 30 and 60 days after the end of the radiation therapy. In this study, 87% of the lesions had complete remission and 13% partial remission to the treatment. Side effects were considered mild according to Veterinary Radiation Therapy Onc ology Group Toxicity criteria, and included erythema, epilation and rhinitis. Radiation Therapy was considered safe for feline cutaneous squamous cell carcinoma, leading to mild side effects and can represent a good therapeutic option.

Distinct types of tumors exhibit differential grade of inflammation and angiogenesis in mice.

Inflammation, angiogenesis and cytokine production are common features of almost, if not all tumors. However, the extent of these processes induced by different types of tumors has not been evaluated. We investigated the growth pattern of the experimental metastatic tumors, B16F10 melanoma, CT26.WT colon and 4T1 mammary cells inoculated in the flank of syngeneic mice and determined the degree of inflammation, angiogenesis, and production level of pro-inflammatory and pro-angiogenic cytokines within the tumors. In addition, we have analyzed vascular changes in the interface between the tumors and the adjacent cutaneous tissue and levels of relevant pro-inflammatory and pro-angiogenic cytokines systemically. The weight of tumors 15 days post-inoculation of 10(6) cells was markedly different. Melanomas were 2 and 10-fold heavier than colon and mammary tumors, respectively. Locally, CT26.WT tumor cells induced more vessels in cutaneous tissue adjacent to the tumors but systemically, the plasma levels of VEGF were higher (approximately 2-fold) in 4T1 tumor-bearing mice compared with the other two tumors. Mammary tumors presented the most prominent inflammatory content as assessed by a range of markers (inflammatory enzymes and cytokines). The vascular index, as determined by the intra-tumor amount of hemoglobin and number of vessels in hot spot areas, was also higher (approximately 2-fold) in melanomas compared with the other two tumors. These findings showing that distinct tumor types determine differential grade of inflammation, angiogenesis and host interaction in mice may provide new insights to tailor differential therapeutic approach based on the status of tumor biomarkers.

Lack of clinical therapeutic benefit of antidepressants is associated overall activation of the inflammatory system.

Despite the evidence of an association between depression and increased inflammatory markers, still little is known in relation to the most severe cases of the disorder i.e., those who fail to respond to antidepressants. We have assessed the cytokine profile and cortisol levels in 21 healthy controls (HC) and 19 medicated patients with depression with treatment-resistance (TRD) moderately ill. As an initial exploratory analysis, we have also related cytokine profile to the patient's clinical treatment outcome after an inpatient admission. Cytokine profile was measured in the serum by the Cytokine Array I kit (Randox). Plasma cortisol was carried out using a commercially available for the IMMULITE system. When compared to healthy controls, depressed patients had higher levels of cortisol, IL-6, IL-10, but lower levels of IL-4 and VEGF. Our exploratory analysis showed subjects who did not go on to respond to the inpatient admission treatment package had lower levels of MCP-1, and a trend toward lower levels of VEGF. Taking together, these data suggest that lack of clinical therapeutic benefit of antidepressants is associated with overall activation of the inflammatory system.

Antidepressants increase human hippocampal neurogenesis by activating the glucocorticoid receptor.

Antidepressants increase adult hippocampal neurogenesis in animal models, but the underlying molecular mechanisms are unknown. In this study, we used human hippocampal progenitor cells to investigate the molecular pathways involved in the antidepressant-induced modulation of neurogenesis. Because our previous studies have shown that antidepressants regulate glucocorticoid receptor (GR) function, we specifically tested whether the GR may be involved in the effects of these drugs on neurogenesis. We found that treatment (for 3-10 days) with the antidepressant, sertraline, increased neuronal differentiation via a GR-dependent mechanism. Specifically, sertraline increased both immature, doublecortin (Dcx)-positive neuroblasts (+16%) and mature, microtubulin-associated protein-2 (MAP2)-positive neurons (+26%). This effect was abolished by the GR-antagonist, RU486. Interestingly, progenitor cell proliferation, as investigated by 5'-bromodeoxyuridine (BrdU) incorporation, was only increased when cells were co-treated with sertraline and the GR-agonist, dexamethasone, (+14%) an effect which was also abolished by RU486. Furthermore, the phosphodiesterase type 4 (PDE4)-inhibitor, rolipram, enhanced the effects of sertraline, whereas the protein kinase A (PKA)-inhibitor, H89, suppressed the effects of sertraline. Indeed, sertraline increased GR transactivation, modified GR phosphorylation and increased expression of the GR-regulated cyclin-dependent kinase-2 (CDK2) inhibitors, p27(Kip1) and p57(Kip2). In conclusion, our data suggest that the antidepressant, sertraline, increases human hippocampal neurogenesis via a GR-dependent mechanism that requires PKA signaling, GR phosphorylation and activation of a specific set of genes. Our data point toward an important role for the GR in the antidepressant-induced modulation of neurogenesis in humans.

Glutathione-related antioxidant defense system in elderly patients treated for hypertension.

The purpose of this study was to analyze glutathione antioxidant defense system in elderly patients treated for hypertension. Studies were carried out in the blood collected from 18 hypertensive and 15 age- and sex-matched controls, all subjects age over 60. Hypertensives were on their usual antihypertensive treatment at the time of blood collection. The concentration of glutathione (GSH) in whole blood and activities of glutathione peroxidase (GPx-1), glutathione transferase (GST), and glutathione reductase (GR) in erythrocytes were measured. The data from patients and controls were compared using independent-samples t test. P value of 0.05 and less was considered statistically significant. We observed increased glutathione-related antioxidant defense in treated hypertensive elderly patients (HT) when compared with healthy controls (C). Mean GSH concentration was significantly higher in HT when compared with C: 3.1 ± 0.29 and 2.6 ± 0.25 mmol/L, respectively, P < 0.001. Mean activity of GR was significantly higher in HT group if compared with C: 83.4 ± 15.25 U/g Hb versus 64.2 ± 8.26 U/g Hb, respectively, P < 0.001. Mean activity of GST was significantly higher in HT group compared with C: 3.0 ± 0.60 mmol CDNB-GSH/mgHb/min and 2.6 ± 0.36 mmol CDNB-GSH/mgHb/min, respectively, P < 0.05. No difference in GPx activity was observed between two groups. These results show that glutathione-related antioxidant defense system was enhanced in elderly hypertensive patients treated for their conditions. This suggests important role of glutathione system in blood pressure regulation. Alterations in concentration and activity of antioxidants observed during antihypertensive medication are likely to be related to the effect of the treatment on NO bioavailability.

Tratamento hipofracionado de radioterapia em felinos portadores de carcinoma epidermóide facial / Hypofractionated radiation therapy for the treatment of feline facial squamous cell carcinoma

Avaliou-se a eficácia do protocolo radioterápico hipofracionado no tratamento de felinos portadores de carcinoma epidermóide facial. Um protocolo hipofracionado de radioterapia foi aplicado em cinco gatos portadores de um ou mais carcinomas epidermóides faciais, em um total de 10 lesões neoplásicas, confirmadas por meio de análise histológica. Duas lesões foram classificadas como T1, quatro como T2, duas como T3 e duas como T4. Os animais foram submetidos a quatro frações radioterápicas de 7,6 a 10gy, com intervalo de uma semana entre elas, utilizando-se um acelerador linear com feixe de elétrons. O acompanhamento dos animais foi realizado semanalmente durante o tratamento e aos 30 e 60 dias após o término da radioterapia. Neste estudo, 40 por cento das lesões resultaram em remissão completa, 40 por cento em remissão parcial e 20 por cento não apresentaram resposta ao tratamento. A taxa de resposta encontrada nessa pesquisa foi baixa, porém o protocolo de hipofracionamento radioterápico foi seguro para gatos com carcinoma epidermóide facial e resultou em efeitos colaterais leves/moderados.

The efficacy of hypofractionated radiation protocol for feline facial squamous cell carcinoma was evaluated. Hypofractionated radiation therapy was applied to five cats showing single or multiple facial squamous cell carcinomas, in a total of ten histologically confirmed neoplastic lesions. Of the lesions, two were staged as T1, four as T2, two as T3, and two as T4. The animals were submitted to four radiation fractions from 7.6 to 10 grays each, with one week intervals. The equipment was a linear accelerator with electrons beam. The cats were evaluated weekly during the treatment and 30 and 60 days after the end of the radiation therapy. In this study, 40 percent of the lesions had complete remission, 40 percent partial remission, and 20 percent did not respond to the treatment. Response rates were lower as compared to other protocols previously used. However, hypofractionated radiation protocol was considered safe for feline facial squamous cell carcinoma.

Antidepressants, but not antipsychotics, modulate GR function in human whole blood: an insight into molecular mechanisms.

Clinical studies have demonstrated an impairment of glucocorticoid receptor (GR)-mediated negative feedback on the hypothalamic-pituitary-adrenal (HPA) axis in patients with major depression (GR resistance), and its resolution by antidepressant treatment. Recently, we showed that this impairment is indeed due to a dysfunction of GR in depressed patients (Carvalho et al., 2009), and that the ability of the antidepressant clomipramine to decrease GR function in peripheral blood cells is impaired in patients with major depression who are clinically resistant to treatment (Carvalho et al. 2008). To further investigate the effect of antidepressants on GR function in humans, we have compared the effect of the antidepressants clomipramine, amytriptiline, sertraline, paroxetine and venlafaxine, and of the antipsychotics, haloperidol and risperidone, on GR function in peripheral blood cells from healthy volunteers (n=33). GR function was measured by glucocorticoid inhibition of lypopolysaccharide (LPS)-stimulated interleukin-6 (IL-6) levels. Compared to vehicle-treated cells, all antidepressants inhibited dexamethasone (DEX, 10-100nM) inhibition of LPS-stimulated IL-6 levels (p values ranging from 0.007 to 0.1). This effect was specific to antidepressants, as antipsychotics had no effect on DEX-inhibition of LPS-stimulated IL-6 levels. The phosphodiesterase (PDE) type 4 inhibitor, rolipram, potentiated the effect of antidepressants on GR function, while the GR antagonist, RU-486, inhibited the effect of antidepressants on GR function. These findings indicate that the effect of antidepressants on GR function are specific for this class of psychotropic drugs, and involve second messenger pathways relevant to GR function and inflammation. Furthermore, it also points towards a possible mechanism by which one maybe able to overcome treatment-resistant depression. Research in this field will lead to new insights into the pathophysiology and treatment of affective disorders.

Vibrational spectroscopy studies on linear polyamines.

Vibrational spectroscopy [both Raman and INS (inelastic neutron scattering)], coupled to quantum mechanical calculations, was used in order to perform a thorough structural analysis of linear polyamines and polynuclear polyamine metal chelates [e.g. with Pt(II) and Pd(II)] with potential anticancer activity. The complementarity of the Raman and INS spectroscopies was exploited in order to gain a better knowledge of the conformational behaviour of these systems. Moreover, the conjugation of the experimental spectroscopic data to the theoretical results allows us to obtain valuable information on the structural preferences of this kind of system, which may lead to the establishment of SARs (structure-activity relationships) ruling their biological activity. Some of the most significant results obtained by the 'Molecular Physical-Chemistry' Research Group of the University of Coimbra (Portugal) are reviewed here.