[Gonococcal pelviperitonitis: a diagnostic challenge].

Neisseria gonorrhoeae is the second most common etiological agent of pelvic inflammatory disease and is currently un-derdiagnosed due to its asymptomatic presentation in 50% of cases. When the disease presents, it may appear in the form of acute abdomen and normal imaging tests, making it a major diagnostic challenge. We present four cases of acute gonococcal peritonitis. The main symptom was acute abdominal pain, and both the gy-necological examination and complementary tests showed normal results. The only notable finding from the laparoscopy was the existence of purulent ascitic fluid. The results of the anatomical and pathological tests were all normal. Endocer-vical and ascitic fluid culture showed infection with N. gonorrhoeae, and in one case, concomitant infection with Chlamydia trachomatis. The definitive treatment applied was intravenous antibiotic therapy. When a sexually active young woman is diagnosed with peritonitis that has no apparent cause, it is important to rule out sexually transmitted diseases.

Exome sequencing of early-onset patients supports genetic heterogeneity in colorectal cancer.

Colorectal cancer (CRC) is a complex disease that can be caused by a spectrum of genetic variants ranging from low to high penetrance changes, that interact with the environment to determine which individuals will develop the disease. In this study, we sequenced 20 early-onset CRC patients to discover novel genetic variants that could be linked to the prompt disease development. Eight genes, CHAD, CHD1L, ERCC6, IGTB7, PTPN13, SPATA20, TDG and TGS1, were selected and re-sequenced in a further 304 early onset CRC patients to search for rare, high-impact variants. Although we found a recurring truncating variant in the TDG gene shared by two independent patients, the results obtained did not help consolidate any of the candidates as promising CRC predisposing genes. However, we found that potential risk alleles in our extended list of candidate variants have a tendency to appear at higher numbers in younger cases. This supports the idea that CRC onset may be oligogenic in nature and may show molecular heterogeneity. Further, larger and robust studies are thus needed to unravel the genetics behind early-onset CRC development, coupled with novel functional analyses and omic approaches that may offer complementary insight.

Ascitis secundaria a chlamydia trachomatis tras procedimiento de reproducción asistida / Ascites secondary to chlamydia trachomatis after assisted reproduction procedure

El desarrollo de ascitis moderada o severa es infrecuente tras una enfermedad inflamatoria pélvica por Chlamydia trachomatis, una de las principales causas de infección de transmisión sexual a nivel mundial. Caso clínico: Paciente de 29 años que tras aborto diferido (gestación tras inseminación artificial) que inicia a las seis semanas con cuadro de dolor abdominal inespecífico y ascitis de predominio linfocitario. El diagnostico se realizo mediante PCR (Werfen®) tanto el liquido ascítico como en exudado endocervical. La paciente recibió tratamiento antibiótico con doxiciclina. Conclusión: Las enfermedades de transmisión sexual deben ser consideradas cuando se realiza un diagnóstico diferencial de una mujer sexualmente activa con dolor abdominal y ascitis, instaurar tratamiento antibiótico y evitar pruebas e intervenciones quirúrgicas innecesarias.

The development of moderate or severe ascites is infrequent after a pelvic inflammatory disease from Chlamydia trachomatis, one of the main causes of sexually transmitted infection worldwide. Clinical case: A 29-year-old patient who, after a delayed abortion (gestation after artificial insemination), started at six weeks with symptoms of non-specific abdominal pain and predominantly lymphocytic ascites. The diagnosis is made by PCR (Werfen®) both the ascitic fluid and the endocervical exudate. The patient received antibiotic treatment with doxycycline. Conclusion: Sexually transmitted diseases should be considered when making a differential diagnosis of a sexually activated woman with abdominal pain and ascites. Establishing antibiotic treatment, and avoiding unnecessary tests and surgical treatments.

Carrier-mediated uptake of L-(+)-lactate in plasma membrane vesicles from rat liver.

Plasma membrane vesicles from rat liver transported L-lactate into the inner vesicular space. Kinetic analysis of L-lactate uptake gave a Km value of approx. 2.9 mM. Selective inhibition was found in a similar pattern to that described for the hepatic lactate carrier. L-Lactate transport was enhanced when a pH gradient was created across the plasma membrane. Vesicles obtained from fasted rats showed a higher uptake of L-lactate than those from fed rats, when incubated with physiological concentrations of L-lactate.

Plasminogen: an important hemostatic parameter in septic patients.

BACKGROUND: Previously we observed in some but not all septic patients a low plasma concentration of plasminogen. OBJECTIVES: To investigate prospectively whether plasma levels of plasminogen or the ratio of plasminogen to alpha-2-antiplasmin have a prognostic value for survival from sepsis and to study the variation of other hemostatic parameters during septicemia. PATIENTS: The study population consisted of 45 consecutive patients with septicemia, 15 non-septic patients from the same intensive care unit and 30 healthy volunteers. MEASUREMENTS AND MAIN RESULTS: Plasminogen concentrations were significantly lower (p < 0.001) in plasma of septic patients (median 0,62 IU/ml range: 0.15-1,06) than in plasma of healthy controls (median 1.00 IU/ml, range: 0.75-1.10) or of non-septic intensive care patients (median 1.00 IU/ml, range: 0.82-1.08). Among the other parameters tested, plasminogen activator inhibitor (PAI-1) antigen concentration and PAI activity were similar in septic and non-septic intensive care patients, but higher than in healthy controls. Concentrations of elastase-alpha-1-protease inhibitor or of thrombin-antithrombin complexes were higher in septic patients than in non-septic intensive care patients or healthy controls. A degraded form of plasminogen of 38 kDa was detected by Western blot analysis in the plasma of septic patients, but not in plasma of non-septic intensive care patients or controls. Plasminogen alone or the ratio of plasminogen to antiplasmin were good markers for survival from septicemia. E.g. for plasminogen at a cut off of 0.65 IU/ml, sensitivity was 90.5% and specificity 66.7%, whereas for the ratio of plasminogen over antiplasmin at a cut off ratio of 0,65 IU/ml, sensitivity was 95.2% and specificity 70.8%. CONCLUSION: Plasminogen or the ratio of plasminogen to antiplasmin are sensitive markers for survival in patients with septicemia.

Epidermal growth factor interferes with the effect of adrenaline on glucose production and on hepatic lipase secretion in rat hepatocytes.

We studied the interaction of epidermal growth factor (EGF) and adrenaline in the control of several metabolic functions in isolated hepatocytes from fed rats. EGF did not modulate glucose release, urea production or hepatic lipase secretion, but interfered with the stimulatory effect of adrenaline on both glucose and urea production and also with the inhibitory effect of this hormone on hepatic lipase secretion. EGF also interfered with the effect of both angiotensin II and vasopressin on glucose release and on hepatic lipase secretion. While the effect of EGF interfering with the action of adrenaline on glucose release was potentiated in the absence of extracellular calcium, the effect on the inhibition of hepatic lipase secretion was abolished. These results suggest that EGF interfered with catecholamine actions in the liver at a site distal from the generation of the calcium signal.

Monoclonal antibody anti-MPO is useful in recognizing minimally differentiated acute myeloid leukaemia.

The enzyme myeloperoxidase (MPO) is the most specific marker of myeloid lineage. The recognition of acute myeloid leukaemia (AML) with minimally differentiation (AML-M0) is established with methods that include myeloid markers CD13/CD33 and detection of MPO in blast cells by immunological techniques or electron microscopy cytochemistry (EM). We have analysed the presence of MPO in leukaemic blast cells by conventional cytochemistry and immunological methods using a monoclonal antibody anti-MPO (CLB-MPO1) in 121 cases of acute leukaemia. The aim of the study was to investigate the sensitivity of this McAb to identify AML-M0, as CD13/CD33 can be expressed in some cases of acute lymphoblastic leukaemia (ALL) and EM cytochemistry is not always available in many laboratories. Anti-MPO was positive in all cases of AML (M1-M5) which were positive by Sudan Black B reaction in similar or higher percentage ratio for each case, although in some of them did not label with CD13/CD33 tested by IF and IPc techniques. Based on the anti-MPO positivity, 5 out of 10 cases called undifferentiated leukaemia (AUL) were reclassified as AML-M0, though 4 cases were CD13/CD33 negative. Furthermore, after analysing the anti-MPO expression among 32 cases of ALL, we had to reclassify four of them as acute biphenotypic leukaemia. We conclude that anti-MPO is a very sensitive and reliable tool in AML diagnosis and has an important role in distinguishing minimally differentiated AML and biphenotypic acute leukaemia from AUL and ALL.

Plasmatic homocysteine response to vitamin supplementation in elderly people.

Homocysteine (Hcy) increase is now widely accepted as a risk factor for vascular disease. The effects of folic acid (FA) and vitamins B12 and B6 in lowering Hcy have been extensively studied, but there is still little data on the response to FA dietary administration. Our purpose was to evaluate the impact of the diet and the degree of response to different doses of pharmacological FA supplementation. In a prospective, randomized, and simple blind study, 50 elderly subjects were given a 400-microg/day FA diet and were randomly assigned to one of the following treatments: Group I = placebo tablet; Group II = tablet containing 1-mg folic acid, 1-mg B12, and 25-mg B6; and Group III = tablet containing 2.5-mg folic acid and same B6 and B12 doses as Group II. Forty-four subjects completed the study, and their plasmas were evaluated. Hcy concentration significantly decreased even in patients with normal basal values, and there were no differences in the response between individuals receiving diet plus placebo and those receiving diet plus pharmacological supplementation. After the treatment, the mean decrease of plasmatic Hcy levels was 10.8 (9.4, 12.5) micromol/l, geometric mean [95% confidence interval (95% CI)], and particularly, the values for Group I were 10.6 (7.4, 14.8) micromol/l. In 31% of the subjects, the post-treatment Hcy levels were less than or = 5 micromol/l. These results show that a special diet, with or without pharmacological FA and B12 and B6 supplementation, significantly decreases the Hcy levels in elderly people. Therefore, a diet with high contents of FA might have an enormous impact on the morbidity and mortality of atherothrombosis.

Heparin cofactor II in diabetic patients.

Levels of heparin cofactor II (HCII) activity and antigen and electrophoretic pattern were studied both in normal subjects and in type I diabetic patients with high and normal levels of glycosylated haemoglobin. There was a significant reduction in HCII activity (83 +/- 7%) in patients with high levels of glycosylated haemoglobin compared with controls (95 +/- 17%; P < 0.001). However, plasma HCII antigen levels were not decreased in these patients.

Unsuspected hyperhomocysteinemia in chronically anticoagulated patients.

Hyperhomocysteinemia is a risk factor for arterial and venous thrombosis. The aim of this study was to evaluate plasmatic homocysteine levels in patients under chronic anticoagulant treatment with dietary restriction of green vegetables. This kind of food is a very important source not only of vitamin K but also of folates, which are involved in Hcy metabolism. It is known that the lower the folate levels, the higher the Hcy concentration, so we suspected that these patients could show hyperhomocysteinemia. A group of patients receiving oral anticoagulant treatment and a restricted diet (Group I, n = 20) was compared with a group of untreated subjects of a similar age that were not on a restricted diet (Group II, n = 35). Group I showed significantly higher levels of plasmatic Hcy and significantly lower levels of serum folate than Group II. Therefore, a diet restricted in vitamin K applied to oral anticoagulated patients could induce an unwanted increase of homocysteine levels.

Validation of an enzyme immunoassay for the determination of total homocysteine in plasma.

In recent years, the determination of homocysteine (Hcy) has become increasingly important, since high levels of Hcy in plasma or serum represent an independent risk factor for occlusive vascular diseases. Nowadays, clinical laboratories use several analytical techniques to measure Hcy, of which high-performance liquid chromatography (HPLC) is the most popular. Recently, assays for Hcy quantification based on enzyme immunoassays (EIA) have become commercially available. Our group carried out the validation of the Axis method and compared results with those obtained by an established HPLC assay. Intra- and inter-assay coefficients of variation were < or = 8.5%. Compared with HPLC, linear regression analysis showed r=0.984, slope=0.952, intercept = 1.24 /mol/l; Bland-Altman procedure, the mean of the difference EIA-HPLC results = 0.5 micromol/l. Our results suggest that Hcy determinations by both methods are equivalent, and that the Axis assay provides reproducible and reliable data.

Cell adhesion response on femtosecond laser initiated liquid assisted silicon surface.

Silicon substrates were irradiated at normal incidence with a femtosecond Ti:sapphire laser (Quatronix, 90 fs pulse duration, 1 kHz repetition rate, M(2) ~ 1.2, maximum energy peak 350 mJ ) operating at a wavelength of 400 nm and focused via a microscope objective (Newport; UV Objective Model, 37x 0.11 N.A.). The laser scanning was assisted by liquids precursors media such as methanol and 1,1,2-trichlorotrifluoroethane. By altering the processing parameters, such as incident laser energy, scanning speed, and different irradiation media, various surface structures were produced on areas with 1 mm(2) dimensions. We analyzed the dependence of the surface morphology on laser pulse energy, scanning speed and irradiation media. Well ordered areas are developed without imposing any boundary conditions for the capillary waves that coarsens the ripple pattern. To assess biomaterial-driven cell adhesion response we investigated actin filaments organization and cell morphological changes following growth onto processed silicon substrates. Our study of bone cell progenitor interaction with laser nanoprocessed silicon lines has shown that cells anchor mainly to contact points along the nanostructured surface. Consequently, actin filaments are stretched towards the 15 µm wide parallel lines increasing lateral cell spreading and changing the bipolar shape of mesenchymal stem cells.

Ultra-fast laser microprocessing of medical polymers for cell engineering applications.

Picosecond laser micromachining technology (PLM) has been employed as a tool for the fabrication of 3D structured substrates. These substrates have been used as supports in the in vitro study of the effect of substrate topography on cell behavior. Different micropatterns were PLM-generated on polystyrene (PS) and poly-L-lactide (PLLA) and employed to study cellular proliferation and morphology of breast cancer cells. The laser-induced microstructures included parallel lines of comparable width to that of a single cell (which in this case is roughly 20µm), and the fabrication of square-like compartments of a much larger area than a single cell (250,000µm(2)). The results obtained from this in vitro study showed that though the laser treatment altered substrate roughness, it did not noticeably affect the adhesion and proliferation of the breast cancer cells. However, pattern direction directly affected cell proliferation, leading to a guided growth of cell clusters along the pattern direction. When cultured in square-like compartments, cells remained confined inside these for eleven incubation days. According to these results, laser micromachining with ultra-short laser pulses is a suitable method to directly modify the cell microenvironment in order to induce a predefined cellular behavior and to study the effect of the physical microenvironment on cell proliferation.

Controversias en el tratamiento de la estrumosis peritoneal: caso clínico y revisión bibliográfica / Controversies in the management of peritoneal strumosis: Case report and literature review

Resumen ANTECEDENTES: el estruma ovárico es un teratoma monodérmico, sumamente complejo, constituido en más de 50% por tejido tiroideo. Una forma infrecuente de manifestación es la estrumosis peritoneal, que consiste en múltiples implantes peritoneales benignos compuestos por estruma ovárico. No existe consenso en relación con el tratamiento; además, la malignidad de la estrumosis peritoneal es tema de controversia. CASO CLÍNICO: paciente nuligesta de 33 años de edad a quien, durante el estudio de esterilidad primaria, se le diagnosticó estruma ovárico derecho. A los tres años postratamiento recurrió en forma de estruma ovárico izquierdo y estrumosis peritoneal (hallazgo intraoperatorio). En ambos casos se efectuó quistectomía laparoscópica, y en la segunda ocasión se practicó, además, la exéresis de los implantes macroscópicos. Después de recibir quimioterapia con 131I (100 mCi) y entrar en remisión completa, se produjo una nueva recidiva cuatro años después; fue necesaria la cirugía citorreductora y un nuevo ciclo de quimioterapia con 131I (100 mCi). En la actualidad la paciente permanece en remisión completa. CONCLUSIONES: ante la escasez de casuística de estrumas ováricos y, sobre todo, de estrumosis peritoneal, resulta imprescindible individualizar el tratamiento en cada paciente, en donde es decisiva la participación de un equipo multidisciplinario. La cirugía citorreductora puede ser una opción terapéutica para favorecer la respuesta al tratamiento ablativo con 131I.

Abstract BACKGROUND: Struma ovarii is a highly specialized monodermic teratoma composed of thyroid tissue in more than 50%. An infrequent form of presentation is peritoneal strumosis, which consists of multiple peritoneal implants of benign mature thyroid tissue. In both situations, there is no consensus about the management of those entities. In addition, malignancy of peritoneal strumosis is controversial. CLINICAL CASE: We report a case of a 33-year-old nulligravid woman that during a sterility study was diagnosed with a right struma ovarii. Three years after treatment, it recurred as a left struma ovarii and a peritoneal strumosis (intraoperative finding). A laparoscopic ovarian cystectomy was performed in both cases, as well as the excision of all macroscopic implants in the second one. She remained in remission after 131I ablative chemotherapy (100 mCi), but a new recurrence took place four years later, requiring a cytoreductive surgery and a second 131I chemotherapy (100 mCi). She is now in complete remission. CONCLUSIONS: Due to the lack of casuistry of struma ovarii and paucity of information, especially in the case of peritoneal strumosis, decisions must be individualized on each patient and must be taken by a multidisciplinary team. Cytoreductive surgery might be an option to improve the response to ablative 131I chemotherapy.

The mechanism of burn-in loss in a high efficiency polymer solar cell.

Degradation in a high efficiency polymer solar cell is caused by the formation of states in the bandgap. These states increase the energetic disorder in the system. The power conversion efficiency loss does not occur when current is run through the device in the dark but occurs when the active layer is photo-excited.

Overview of Anticoagulant Activity of Sulfated Polysaccharides from Seaweeds in Relation to their Structures, Focusing on those of Green Seaweeds

The anticoagulant behavior of sulfated polysaccharides from seaweeds is reviewed based on their chemical structures. Analysis of the literature suggested that the driving force for the formation of the sulfated polysaccharide/protein complex is the non-specific polar interaction between the negatively and positively charged groups in the polysaccharide and protein, respectively and that the complex is further stabilized by short-range interactions. The polysaccharide binding site should be able to go through the following conformational steps in the formation of the complex: random coil-->ordered conformation--> low distortion of this conformation to form a complementary fitting structure with the protein backbone. The sulfated monosaccharide units with the highest potential for anticoagulant activity should have two sulfate groups and a glycosidic linkage on the pyranose ring with C-2, C-3 and C-4 in 2S, 3R, 4R or 2R, 3S, 4S configurations for galactose, fucose and arabinose and 2S, 3S, 4R, for rhamnose. Three distributions of these substituents appear: 3-linked 2,4-disulfated units, 4-linked 2,3-disulfated units and 2-linked 3,4-disulfated residues. These types of units have the possibility, through the equilibrium of the chair conformations, to place their sulfate groups in adequate special positions to interact with basic groups of the protein. The anticoagulant activity is mainly attributed to thrombin inhibition mediated by antithrombin and/or heparin cofactor II, with different effectivenesses depending of the compound. Other mechanisms are also proposed and these differences could be attributed to the diversity of structures of the polysaccharides evaluated and to the fact that one compound may have more than one target protease.

Regulation of the antibody response to sheep erythrocytes by monoclonal IgG antibodies.

Monoclonal antibodies directed against sheep erythrocytes of the isotypes IgG1, IgG2b and IgG2a were used to analyze the specificity of antibody-induced suppression of the immune response. It was first shown that all monoclonals reacted against different antigenic determinants and they all suppressed the immune response to sheep erythrocytes when given shortly after the antigen to more than 50% as compared to 90-96% inhibition obtained with a polyclonal antiserum. Increasing the doses of monoclonals did not increase suppression. However, two different monoclonals administered together caused an additive, but not a synergistic inhibitory effect. No enhancement of the immune response was observed with any of the Ig classes tested. These findings show that four different antigenic determinants on sheep erythrocytes induced the synthesis of corresponding antibodies, with little or no signs of a dominant determinant. Passively administered monoclonal antibodies, even at supraoptimal doses, never suppressed the immune response to the same extent as a polyclonal antiserum, suggesting that each monoclonal only suppressed the synthesis of the corresponding antibody and did not affect antibody synthesis to other determinants.

[Impact of vaccination against Haemophilus influenzae type b in Cuba]. / Impacto de la vacunación contra Haemophilus influenzae tipo b en Cuba.

OBJECTIVE: To assess the effectiveness of mass vaccination in Cuba of children under 2 years of age against Haemophilus influenzae type b (Hib), the most common causative pathogen of bacterial meningitis. METHODS: The availability of effective Hib conjugate vaccines led to a nationwide vaccination program in 1999 targeting all children under 2 years of age, with a 97% coverage rate achieved. To assess the program's impact, data from 1998 and 1999 from the National Bacterial Meningitis Reporting System were used. RESULTS: Vaccination efficacy was estimated at 99%. The overall incidence of Hib meningitis declined 46.1%, from 1.3 to 0.6 cases per 100,000 population. The greatest overall reduction, of 56.1%, occurred among children under 5 years of age. Among children under 1 year of age, the reduction was 70.5%, and among the rest of the age groups of children under 5, incidence decreased between 25.9% and 49.6%. In the group targeted for vaccination, incidence decreased 61.1%. Among children in the target group who contracted Hib meningitis, only 8 cases (24.2%) had been vaccinated, most with a single dose applied 1 month before becoming ill. CONCLUSIONS: Hib vaccination of all children under 2 years of age in Cuba greatly reduced the incidence of Hib meningitis, as measured by the National Bacterial Meningitis Reporting System.

The early stimulation of glycolysis by epidermal growth factor in isolated rat hepatocytes is secondary to the glycogenolytic effect.

We have studied the relationship between the effect of epidermal growth factor (EGF) on glycogen metabolism and its effect on glycolysis, in rat hepatocyte suspensions. Although 10 nM glucagon or 10 microM adrenaline increased glycogen degradation by more than 120%, 10 nM EGF increased glycogenolysis by less than 20% in hepatocytes incubated in glucose-free medium. Both glucagon and adrenaline increased phosphorylase a activity by more than 130%; EGF increased this activity by about 30%. Under basal conditions, 65% of the glucosyl residues were released as free glucose and about 30% ended up as C3 molecules (lactate and pyruvate). Both glucagon and adrenaline decreased the proportion of glucosyl units that rendered glycolysis end-products (to 2% for glucagon and 6% for adrenaline) and increased the proportion that ended up as free glucose (to 94% and 88% of the glucosyl residues for glucagon and adrenaline respectively). EGF increased the production of both free glucose and lactate+pyruvate, but the proportion of glucosyl residues that ended up as free glucose or glycolysis end-products was unchanged. In glycogen-depleted hepatocytes incubated in the presence of 25 mM glucose, EGF affected neither glycogen deposition nor glycolysis. EGF increased cytosolic free Ca2+, and neomycin decreased both the Ca2+ signal and the glycogenolytic effect. In conclusion, our results indicate that the effect of EGF on glycolysis is secondary to the Ca(2+)-mediated stimulation of glycogenolysis in rat hepatocyte suspensions.

Typing and susceptibility to penicillin of Neisseria meningitidis isolated from patients in Cuba (1993-1999).

The susceptibility to penicillin of 111 Neisseria meningitidis strains was assessed by the agar-dilution procedure and serosubtypes were determined by a whole-cell enzyme-linked immunoassay using monoclonal antibodies reagents. Thirty-five isolates showed reduced sensitivity to penicillin (MIC > or = 0.1 mg/l and < or = 1 mg/l) and no resistant strains were detected. The most common phenotype was B:4:P1.15 (77.5%) and a rising trend of non-typeable and non-subtypeable strains was detected. The increase in levels of minimal inhibitory concentrations of meningococci to penicillin gives cause for concern and the increase in non-typeable and non-subtypeable isolation demand the use of molecular biology techniques for their typing.