Cytosolic superoxide dismutase activity after photodynamic therapy, intracellular distribution of Photofrin II and hypericin, and P-glycoprotein localization in human colon adenocarcinoma

In photodynamic therapy (PDT), a tumor-selective photosensitizer is administered and then activated by exposure to a light source of applicable wavelength. Multidrug resistance (MDR) is largely caused by the efflux of therapeutics from the tumor cell by means of P-glycoprotein (P-gp), resulting in reduced efficacy of the anticancer therapy. This study deals with photodynamic therapy with Photofrin II (Ph II) and hypericin (Hyp) on sensitive and doxorubicin-resistant colon cancer cell lines. Changes in cytosolic superoxide dismutase (SOD1) activity after PDT and the intracellular accumulation of photosensitizers in sensitive and resistant colon cancer cell lines were examined. The photosensitizers' distributions indicate that Ph II could be a potential substrate for P-gp, in contrast to Hyp. We observed an increase in SOD1 activity after PDT for both photosensitizing agents. The changes in SOD1 activity show that photodynamic action generates oxidative stress in the treated cells. P-gp appears to play a role in the intracellular accumulation of Ph II. Therefore the efficacy of PDT on multidrug-resistant cells depends on the affinity of P-gp to the photosensitizer used. The weaker accumulation of photosensitizing agents enhances the antioxidant response, and this could influence the efficacy of PDT.     

Morphology and the central nervous system of <Emphasis Type="Italic">Eratigena atrica</Emphasis> affected by a complex anomaly in the anterior part of the prosoma

Spider embryogenesis is affected by a range of environmental factors. Any sudden, drastic change in the environment may impair spider development, leading to various body deformities. In the present study, we analyze changes in the morphology and structure of the central nervous system of an Eratigena atrica larva, obtained in a teratological experiment in which embryos were exposed to alternating temperatures of 14 and 32 °C for the first 10 days. The studied larva had three pedipalps on the right side of the prosoma (polymely), two of which were fused along their entire length (total heterosymely). In addition, there was a short, club-shaped stump between the pedipalps. Histological analysis confirmed major changes in the structure of the subesophageal ganglion, i.e., the fusion of all three ganglia of pedipalps.     

Factors Associated with the Time of Admission among Notified Dengue Fever Cases in Region VIII Philippines from 2008 to 2014

In cases of Dengue fever, late hospital admission can lead to treatment delay and even death. In order to improve early disease notification and management, it is essential to investigate the factors affecting the time of admission of Dengue cases. This study determined the factors associated with the time of admission among notified Dengue cases. The study covered the period between 2008 and 2014 in Region VIII, Philippines. The factors assessed were age, sex, hospital sector, hospital level, disease severity based on the 1997 WHO Dengue classification, and period of admission (distinguishing between the 2010 Dengue epidemic and non-epidemic time). We analysed secondary data from the surveillance of notified Dengue cases. We calculated the association through chi-square test, ordinal logistic regression and linear regression at p value < 0.05. The study included 16,357 admitted Dengue cases. The reported cases included a majority of children (70.09%), mild cases of the disease (64.00%), patients from the public sector (69.82%), and non-tertiary hospitals (62.76%). Only 1.40% of cases had a laboratory confirmation. The epidemic period in 2010 comprised 48.68% of all the admitted cases during this period. Late admission was more likely among adults than children (p<0.05). The severe type of the disease was more likely to be admitted late than the mild type (p<0.05). Late admission was also more likely in public hospitals than in private hospitals (p<0.05); and within tertiary level hospitals than non-tertiary hospitals (p<0.05). Late admission was more likely during the non-epidemic period than the 2010 epidemic period (p<0.05). A case fatality rate of 1 or greater was significantly associated with children, severe diseases, tertiary hospitals and public hospitals when admitted late (p<0.05). Data suggests that early admission among child cases was common in Region VIII. This behavior is encouraging, and should be continued. However, further study is needed on the late admission among tertiary, public hospitals and non-epidemic period with reference to the quality of care, patient volume, out of pocket expense, and accessibility We recommend the consistent use of the 2009 WHO Dengue guidelines in order to standardize the admission criteria and time across hospitals.     

Estimation of cell membrane properties and erythrocyte red-ox balance in patients with metabolic syndrome

Metabolic syndrome (MS) is associated with occurrence of the many cardiovascular risk factors such as atherogenic dyslipidemia, visceral fat distribution, arterial hypertension and pro-thrombotic and pro-inflammatory status. In our study the effect of disorders that appear in MS on red-ox balance and erythrocyte cell membrane properties were estimated. The study comprised 50 patients with diagnosed MS and in 25 healthy subjects. Content of thiobarbituric acid reactive substances (TBARS) and catalase, superoxide dismutase and glutathione peroxidase activity were estimated in red blood cells. Moreover, conformation status of membrane proteins, membrane fluidity and osmotic fragility were evaluated. MS was found to manifest: (1) the increase of the concentration of TBARS in erythrocytes with no statistically significant differences in antioxidant enzymes activity, (2) disorders in the structure of erythrocyte cytoskeleton proteins, (3) the increase in membrane lipids fluidity at the depth of 5th and 12th carbon atom of fatty acid hydrocarbon chain and significantly decreased fluidity at the depth of 16th carbon atom, (4) increased erythrocyte osmotic fragility.     

Damage to the structure of erythrocyte plasma membranes in patients with type-2 hypercholesterolemia

BACKGROUND: Hypercholesterolemia may decrease the deformability of red blood cells which impairs their hemorheological behavior and promotes atherosclerosis.The study involved 60 hypercholesterolemic patients and 30 healthy individuals as the control group.METHODS: The membrane fluidity of erythrocytes was estimated by a spin-label method (5-doxylstearic acid (5-DSA)). The ratio of weakly to strongly (W/S) immobilized residues of erythrocyte membrane-bond maleimide-tempo spin label was studied in oxidative damage to membrane protein. Damage to erythrocyte proteins was also indicated by means of Na(+) K(+) ATPase activity.RESULTS: The membranes of hyperlipidemia (hlp) patients contain larger concentrations of cholesterol 2.16+/-0.24 than do those of the normolipemic individuals 0.31+/-0.24 (P<0.001). The level of Na(+) K(+) ATPase in the erythrocyte membrane from the control group was higher 103.4+/-1.3 (nmolPi/(mgproteinsh)) than in the patient group 93.6+/-3.2 (nmolPi/(mgproteinsh)) (P<0.001). The order parameter S 5-DSA in the control group was 0.745+/-0.009 and in hlp patients was 0.755+/-0.009 (P<0.001). The W/S ratio in the control group amounted to 2.00+/-0.09 and in the hlp patient group was 2.50+/-0.11 (P<0.001).CONCLUSION: Type-2 hypercholesterolemia causes changes in the structure and fluidity of erythrocyte plasma membranes since the excess of cholesterol affects the normal rheology of blood through its interaction with erythrocytes. It also impairs the function and structure of plasma membrane proteins.     

Effects of simvastatin and pravastatin on peroxidation of erythrocyte plasma membrane lipids in patients with type 2 hypercholesterolemia

Since hypercholesterolemia directly modifies the composition of erythrocytes plasma membrane, the influence of statins on erythrocytes has been researched. The beneficial effects of statins on clinical events may involve mechanisms that modify endothelial dysfunction, plaque stability, thrombus formation and inflammatory responses. The aim of the study was to evaluate the hypolipemic efficacy and effects of pravastatin and simvastatin on erythrocyte membrane fluidity and damage of erythrocytes in patients with type 2 hypercholesterolemia in comparison with a control group of healthy subjects. The study involved 53 patients affected by type 2 hypercholesterolemia (mean age, 53.3 +/- 10.3) with initial total serum cholesterol (TC) levels > 250 mg/dL, LDL-cholesterol (LDL-C) levels > 170 mg/dL, and triglycerides (TG) levels < 400 mg/dL. The control group consisted of 30 healthy individuals (mean age 56.9 +/- 6.3). Statins were given for 12 weeks. The dosages for oral administration of simvastatin and pravastatin were 20 mg/day. Laboratory tests were carried out before and after 4 and 12 weeks of the pharmacological treatment. The damage to plasma membrane of erythrocytes was measured on the basis of lipid peroxidation. The fluidity of plasma membrane of erythrocytes was determined by electron paramagnetic resonance (EPR) spectroscopy, using two spin labels: 5-DSA and 16-DSA. The cholesterol level in the membrane of red blood cells was estimated. Simvastatin and pravastatin reduced the total cholesterol concentration and LDL-cholesterol in plasma, as well as the cholesterol concentration in erythrocytes membranes. Hypercholesterolemia induced changes in the basic properties of human erythrocyte plasma membrane, including its fluidity and the intensity of lipid peroxidation. These results indicate that the simvastatin and pravastatin therapy reverses the alteration in the erythrocyte plasma membrane properties.     

Putative gamma-conotoxins in vermivorous cone snails: the case of Conus delessertii

Peptide de7a was purified from the venom of Conus delessertii, a vermivorous cone snail collected in the Yucatan Channel, Mexico. Its amino acid sequence was determined by automatic Edman degradation after reduction and alkylation. The sequence shows six Cys residues arranged in the pattern that defines the O-superfamily of conotoxins, and several post-translationally modified residues. The determination of its molecular mass by means of laser desorption ionization time-of-flight mass spectrometry (average mass, 3170.0 Da) confirmed the chemical data and suggested amidation of the C-terminus. The primary structure (ACKOKNNLCAITgammaMAgammaCCSGFCLIYRCS*; O, hydroxyproline; gamma, gamma-carboxyglutamate; *, amidated C-terminus; calculated average mass, 3169.66 Da) of de7a contains a motif (gammaCCS) that has previously only been found in two other toxins, both from molluscivorous cone snails: TxVIIA from Conus textile and gamma-PnVIIA from Conus pennaceus. These toxins cause depolarization and increased firing of action potentials in molluscan neuronal systems, and toxin gamma-PnVIIA has been shown to act as an agonist of neuronal pacemaker cation currents. The similarities to toxins TxVIIA and gamma-PnVIIA suggest that peptide de7a might also affect voltage-gated nonspecific cation pacemaker channels.     

Selective mirror-image reversal of ciliary patterns inTetrahymena thermophila homozygous for ajanus mutation

Summary The development of the oral apparatus (OA) and of neighboring ciliary structures ofTetrahymena thermophila was analyzed in cells homozygous for ajanus (jan A) mutation plus a recessiveenhancer of janA (eja). Such cells frequently possess two OAs located on opposite sides of the cell, a primary (1°) OA previously reported to be normal, and a secondary (2°) OA previously reported to express a mirror-reversal of right-left asymmetry. This study confirms the reality of a reversal in the gross orientation of membranelles in most developing 2° OAs. It also shows that there is a reversal of asymmetry in the pattern of resorption of basal bodies of ciliary rows adjacent to the 2° OA, and in the arrangement of basal-body couplets making up the portion of the apical crown of the cell situated close to the 2° OA. However, the locations at which membranelles of the 2° OA become modified during late phases of oral development remain normal, so that membranelles of 2° OAs are superimposable on those of 1° OAs. In addition, the membranelles of 2° OAs frequently undergo a rotation during the final phases of oral development, so that even their spatial orientation becomes normal. This mixture of reversed and normal features can be accounted for by postulating a superimposition of a reversed largescale asymmetry on a normal local asymmetry of ciliary units. This postulate predicts that no single mutation can bring about a complete mirror-image reversal of ciliary patterns.1° OAs appear normal by light microscopy. However, detailed analysis of SEM, preparations of isolated 1° OAs indicate subtle abnormalities of basal body arrangement in some of these OAs.     

Organization and ultrastructure of the nervous system in Catenulida (Turbellaria)

Summary The fine structure of the nervous system of lower fresh-water Turbellaria was investigated. This system consists of a brain, short nerve trunks and a network of subepithelial nerve cells. The brain structure shows ganglion cells and their proccesses, forming a neuropil. The ganglion cells are most probably unipolar. The perikaryon contains numerous ribosomes, few mitochondria, and golgi complexes. Thus it corresponds structurally to neuroblasts of higher animals. The neurites contain mitochondria, neurotubules, and empty or dense core vesicles. All (inStenostomum sp.) or some of the nerve cells (inCatenula sp.) have neurosecretory vesicles.