Toxicity assessment of biological suspensions using the dielectric impedance spectroscopy technique.

PURPOSE: This article studies the variation of the electromagnetic parameters of a suspension of zebrafish (Danio rerio) embryos to assess its potential applications to toxicological and biomedical research areas. MATERIALS AND METHODS: For this purpose, the dielectric impedance spectroscopy technique is applied to a modified coaxial line enclosing the biological suspension to be characterized in the frequency range from 100 kHz to 100 MHz. The electrical parameters of the suspension under test were obtained by fitting the impedance spectra to the resulted from the simulation of the test fixture using finite elements (FE). RESULTS: Variation of the complex permittivity of the suspensions makes possible to identify viable and non-viable embryos after a toxic exposure, as well as different stages during the blastula period of embryonic development of the zebrafish. CONCLUSIONS: The approach presented here, combining experimental and simulation techniques, may provide a basis for a non-invasive method to assess toxicity in any biological suspension.

Elastic energy of the discocyte-stomatocyte transformation.

The aim of this study is to calculate the membrane elastic energy for the different shapes observed in the discocyte-stomatocyte sequence. This analysis can provide a better quantitative understanding of the hypothesis put forward over the last decades to explain how red blood cells produce and maintain their typical shape. For this purpose, we use geometrical models based on parametric equations. The energy model considered for the elastic properties of RBC membrane includes the local and nonlocal resistance effects of the bilayer to bending. In particular, the results confirm the discocyte as the lowest energy value configuration among the sets of different red blood cell deformations considered in the sequence.

Sampling methodologies for epidemiologic surveillance of men who have sex with men and transgender women in Latin America: an empiric comparison of convenience sampling, time space sampling, and respondent driven sampling.

Alternatives to convenience sampling (CS) are needed for HIV/STI surveillance of most-at-risk populations in Latin America. We compared CS, time space sampling (TSS), and respondent driven sampling (RDS) for recruitment of men who have sex with men (MSM) and transgender women (TW) in Lima, Peru. During concurrent 60-day periods from June-August, 2011, we recruited MSM/TW for epidemiologic surveillance using CS, TSS, and RDS. A total of 748 participants were recruited through CS, 233 through TSS, and 127 through RDS. The TSS sample included the largest proportion of TW (30.7 %) and the lowest percentage of subjects who had previously participated in HIV/STI research (14.9 %). The prevalence of newly diagnosed HIV infection, according to participants' self-reported previous HIV diagnosis, was highest among TSS recruits (17.9 %) compared with RDS (12.6 %) and CS (10.2 %). TSS identified diverse populations of MSM/TW with higher prevalences of HIV/STIs not accessed by other methods.

Health care utilization and costs of a support program for patients living with the human immunodeficiency virus and tuberculosis in Peru.

OBJECTIVE: To evaluate a support program for patients co-infected with the human immunodeficiency virus and tuberculosis in terms of its impact on clinical outcomes and resource utilization. METHODS: We compared co-infected patients receiving Community-Based Accompaniment with Supervised Antiretrovirals (CASA) with matched patients receiving standard of care (control group) in two health districts of Lima, Peru. We recorded clinical outcomes, costs of the intervention, and health care utilization by each patient during 24 months of follow-up. RESULTS: There were 33 patients in each group, representing 58.0 person-years (py) in the CASA group and 45.6 py in the control group. At 24 months of follow-up, the CASA group had a lower hazard of dying or defaulting from treatment (HR adj 0.34, 95%CI 0.12-0.98), experienced fewer hospital days (IRR adj 0.37, 95%CI 0.14-0.99) and had fewer out-patient visits (IRR adj 0.75, 95%CI 0.63-0.89). Assigning costs to significantly different measures of health care utilization using WHO-CHOICE (World Health Organization-Choosing interventions that are cost effective) data, CASA was associated with savings of US$551/py. Considering intervention costs of US$2097/py, the net costs of CASA were US$1546/py. CONCLUSIONS: Our intervention was associated with clinical improvements and reduced health care utilization, which significantly offset the cost of the intervention over 2 years of follow-up.

HLA homozygous stem cell lines derived from human parthenogenetic blastocysts.

Individual HLA homozygous parthenogenetic human stem cell (hpSC-Hhom) lines have the potential for cell-based therapy in a significant number of individuals, provided the HLA haplotype is prevalent. We report the successful derivation of four stable hpSC-Hhom lines from both HLA homozygous and HLA heterozygous donors. Of these, the hpSC-Hhom-4 line carries the HLA haplotype found most commonly within the U.S. population, and is shared by different racial groups. These hpSC-Hhom lines demonstrate typical human embryonic stem cell morphology, expressing appropriate stem cell markers and possessing high levels of alkaline phosphatase and telomerase activity. Additionally, injection of these cell lines into immunodeficient animals leads to teratoma formation. G-banded karyotyping demonstrates a normal 46,XX karyotype in lines hpSC-Hhom-1 and hpSC-Hhom-4, and chromosomal anomalies in lines hpSC-Hhom-2 and hpSC-Hhom-3, both derived from the same donor. HLA genotyping of all four hpSC-Hhom lines demonstrates that they are HLA homozygous. Furthermore, in the case of HLA heterozygous donors, the hpSC-Hhom lines inherit the haplotype from only one of the donor's parents. Single-nucleotide polymorphism (SNP) data analysis suggests that hpSC-Hhom lines derived from HLA heterozygous oocyte donors are homozygous throughout the genome as assessed by SNP analysis. The protocol used for deriving these HLA homozygous stem cell lines minimizes the use of animal-derived components, which makes them more appealing for potential clinical application.

Electric field distribution and energy absorption in anisotropic and dispersive red blood cells.

We have studied the influence of the anisotropic and dispersive nature of the red blood cell structure on the energy absorption and electric field distribution within the cell exposed to electromagnetic fields of frequencies in the range from 50 kHz to 10 GHz. For this purpose we have generated a realistic model of a multilayered erythrocyte cell from a set of parametric equations in terms of Jacobi elliptic functions. The effect of dipole relaxations and anisotropic conductivities is taken into account in the dispersion equations for the conductivity and permittivity of each layer (cytoplasmic and extra-cellular bound waters, membrane, cytoplasm and external medium). Using a finite element numerical technique, we have found that the electric field distribution and the energy absorbed in the membrane show well-defined maxima for both normal and parallel orientations of the external field with respect to the symmetry axis of the cell. The normal and tangential conductivities and permittivities of the membrane are shown to be responsible for the different peak amplitudes and frequency shifts of the maxima. A previously unnoticed effect is that the cell shape combined with the dispersion of the membrane permittivity and the influence of bound water layers leads to a very high amplification factor (greater than 300) of the electric field in the membrane at frequencies in the megahertz range.

Dielectric characterization of bacterial cells using dielectrophoresis.

Measurements of dielectrophoretic collection spectra of Escherichia coli and Staphylococcus aureus suspensions are used for obtaining dielectric characteristics of both types of bacteria. The experiments are interpreted using a numerical method that models the cells as compartmented spherical or rod-like particles. We show the usefulness of this simple method to extract significant information about the electrical properties of Gram-negative and -positive bacteria.

Analysis of the electric field induced forces in erythrocyte membrane pores using a realistic cell model.

We calculate the induced electric stress forces on transient hydrophobic pores in the membrane of an erythrocyte exposed to an electric field. For this purpose, we use a finite element numerical technique and a realistic shape for the biconcave erythrocyte represented by a set of parametric equations in terms of Jacobi elliptic functions. The results clearly show that the electrical forces on the base and sidewalls of the pore favour the opening of the pore. A comparison of the force densities obtained for an unstretched flat membrane and for the realistic erythrocyte model shows that the thinning and curvature of the membrane cannot be neglected. We also show that the pore deformation depends strongly on the orientation of the pore with respect to the external field, and in particular is very small when the field is tangent to the membrane surface.

Modeling normal and altered human erythrocyte shapes by a new parametric equation: application to the calculation of induced transmembrane potentials.

We present simple parametric equations in terms of Jacobi elliptic functions that provide a realistic model of abnormal variations in size which maintain the biconcave shape of a normal erythrocyte (anisocytosis) and abnormal variations in shape which maintain the original volume of the erythrocyte (poikilocytosis), as well as continuous deformations from the normal to the altered shapes. We illustrate our results with parameterizations of microcytes, macrocytes, and stomatocytes, and we apply these parameterizations to the numerical calculation of the induced transmembrane voltage in microcytes, macrocytes, and stomatocytes exposed to an external electromagnetic field of 1800 MHz.

A PDA-based instructional tool to monitor students' cardiac auscultation during a medicine clerkship.

Cardiac auscultation is an important skill for medical students to master but students' exposure to cardiac auscultation is often unmonitored. The objective of this study was to gather data at the point of care about students' cardiac auscultation experience on a required medicine rotation using a Personal Digital Assistant (PDA) 'murmur form'. During an eight-month period, 120 M3 students used the authors' PDA-based learning tool to record information on 940 heart sounds and murmurs. Some 93% of all heart sounds/murmurs reported by students were verified by either a faculty member (56%) or a supervising resident (43%). A PDA can be a useful tool to monitor students' experiences of cardiac auscultation and to track direct observation of such skills by faculty or residents. Medical students are eager to use technology at the point of care to practice their clinical skills.

Transmembrane voltage induced on altered erythrocyte shapes exposed to RF fields.

In this article, the transmembrane voltage induced on erythrocyte, codocyte, ovalocyte and spherocyte cell models exposed to a linearly polarised electromagnetic plane wave of frequency 1800 MHz is calculated. For this purpose, a finite element (FE) numerical technique with adaptive meshing is used. The results show that the value of the induced voltage on the original erythrocyte shape is higher than the one observed on the rest of the altered cell geometries studied. The erythrocyte shape and the membrane electric permittivity are shown to play a fundamental role on the values of the induced transmembrane voltage.

Modelling the internal field distribution in human erythrocytes exposed to MW radiation.

This paper studies the internal electric field distribution in human erythrocytes exposed to MW radiation. For this purpose, an erythrocyte cell model is exposed to linearly polarized electromagnetic (EM) plane waves of frequency 900 MHz and the electric field within the cell is calculated by using a finite element (FE) technique with adaptive meshing. The results obtained show the dependence of the induced electric field distribution on the main modelling parameters, i.e., the electrical properties (permittivity and conductivity) of the membrane and cytoplasm and the orientation of the cell with respect to the applied field. It is found that for certain orientations, the field amplification within the membrane of the erythrocyte shape cell can be higher than the one observed in an equivalent simple spheroidal geometry cell, commonly used in bioelectromagnetism. The present work shows that a better insight of the interaction of electromagnetic fields with basic biological structures is obtained when the most possible realistic cell shape is used.

A study of the electric field distribution in erythrocyte and rod shape cells from direct RF exposure.

This paper shows the importance of using realistic cell shapes with the proper geometry and orientation to study the mechanisms of direct cellular effects from radiofrequency (RF) exposure. For this purpose, the electric field distribution within erythrocyte, rod and ellipsoidal cell models is calculated by using a finite element technique with adaptive meshing. The three cell models are exposed to linearly polarized electromagnetic plane waves of frequencies 900 and 2450 MHz. The results show that the amplification of the electric field within the membrane of the erythrocyte shape cell is more significant than that observed in other cell geometries. The results obtained show the dependence of the induced electric field distribution on frequency, electrical properties of membrane and cytoplasm and the orientation of the cell with respect to the applied field. The analysis of the transition of an erythrocyte shape to an ellipsoidal one shows that a uniformly shelled ellipsoid model is a rough approximation if a precise simulation of bioeffects in cells is desired.

Long-term follow-up of a randomized prospective trial comparing a standard 81 mg dose of intravesical bacille Calmette-Guérin with a reduced dose of 27 mg in superficial bladder cancer.

OBJECTIVES: To determine the efficacy of a three-fold reduced dose (RD, 27 mg) of intravesical bacille Calmette-Guérin (BCG) against the standard dose (81 mg) in patients with superficial bladder cancer, assessing recurrence, progression and differences in toxicity. PATIENTS AND METHODS: Five hundred patients with superficial bladder cancer (Ta, T1, Tis) were enrolled and randomly assigned to be treated after transurethral resection of all visible lesions with intravesical BCG Connaught strain (weekly x six and thereafter fortnightly x six) either with the standard or RD instillation. RESULTS: All but one of the 500 patients were evaluable for efficacy and toxicity (252 in the standard arm and 247 in the RD arm). The median follow-up was 69 months (maximum 104); 71 (28%) patients in the standard arm and 76 (31%) in the RD arm developed recurrences; the median time to recurrence has not yet been attained, but at 5 years the mean (sd) percentage of recurrence-free patients was 70.5 (3.12) and 70.4 (3.1) for the standard and RD arms, respectively. In patients presenting with multifocal tumours, the standard dose was more effective against recurrences than the RD (P=0.0151). In those with G3 and high-risk tumours overall, the superiority of the standard dose was marginal (P=0.060 and P=0.082). Twenty-nine (11.5%) tumours in the standard arm and 33 (13.3%) in the RD arm progressed to invasive disease; the median time to progression has not yet been attained, but the percentage of progression-free patients at 5 years was 88.8 (2.23) and 86.9 (2.31) for the standard and RD arms, respectively. The standard dose was more effective than the RD against progression only in patients with multifocal disease (P=0.048). Twelve (4.8%) cystectomies were performed in the standard and 15 (6.1%) in the RD arm. Currently, 106 (21.2%) patients have died, but only 38 (7.6%) from bladder cancer, i.e. 20 (7.9%) in the standard and 18 (7.5%) in RD arm. Overall the disease-specific death rate was lower for those patients who completed the scheduled treatment. The cause-specific survival at 5 years did not differ between the arms (P=0.76) but there was a trend toward better cause-specific survival for patients with multifocal tumours in the standard arm. Toxicity differed between the arms, significantly more patients having no toxicity in the RD arm, and fewer having delayed instillations or withdrawing. However, severe systemic toxicity occurred even in patients treated with the RD, in a similar proportion to those receiving the standard dose. CONCLUSION: Overall, the RD gave similar results for recurrence and progression but with significantly less toxicity. However, patients with multifocal tumours fared better with the standard dose and there was a trend towards better recurrence rates in patients with high-risk tumours. We recommend continuing to use the standard dose for high-risk tumours, while we consider the reduced dose safe and effective for intermediate-risk lesions and for maintenance schedules.

Biexponential model for predicting weight loss after gastric surgery for obesity.

BACKGROUND: Following gastric restrictive surgery, morbidly obese patients rarely achieve their ideal body weight defined by Metropolitan Life tables. The final body weight will depend on the initial body composition because there will be greater weight loss from fat than lean body mass. The purpose of this study was to develop a mathematical model that accurately estimates the rate and extent of weight loss following gastric bypass surgery. METHODS: Patients underwent gastric bypass followed by intensive medical therapy and serial bioelectrical impedance analysis (BIA) body composition measurements. Differential equations were derived to model weight loss. RESULTS: Weight loss in the fat and lean body compartments followed monoexponential decay kinetics with differing rate constants. Total body weight loss (W(T)) at time t was W(T) = k(f)(k(f) - k(l)) (W(f(o))e(-k(f)t) + W(l(o))e(-k(l)t)), where W(fo) and W(lo) are the initial fat and lean body masses determined by BIA and k(f) and k(l) are the rate constants for the fat and lean compartments, respectively. Following surgically induced weight loss, k(f) = 7.61 +/- 1.27 x 10(-2), and k(l) = -0.93 +/- 0.13 x 10(-2), with the ratio of residual sum of the squares to the total sum of the squares of 98.8%. CONCLUSION: Accurate prediction of weight loss depends on the initial fat and lean compartment mass since each of these loses weight at a different rate and to a different extent. When these effects are accounted for, the total body weight loss can be accurately predicted for any given time following surgery.

Analysis of the influence of the cell geometry, orientation and cell proximity effects on the electric field distribution from direct RF exposure.

This paper shows the importance of using a cell model with the proper geometry, orientation and internal structure to study possible cellular effects from direct radiofrequency exposure. For this purpose, the electric field intensity is calculated, using the finite element numerical technique, in single- and multilayer spherical, cylindrical and ellipsoidal mammalian cell models exposed to linearly polarized electromagnetic plane waves of frequencies 900 and 2450 MHz. An extensive analysis is performed on the influence that the cell geometry and orientation with respect to the external field have in the value of the electric field induced in the membrane and cytoplasm. We also show the significant role that the cytoplasmic and extracellular bound water layers play in determining the electric field intensity for the cylindrical and ellipsoidal cell models. Finally, a study of the mutual interactions between cells shows that polarizing effects between cells significantly modify the values of field intensity within the cell.

Determinación de la permitividad y conductividad eléctricas a la frecuencia de microondas de varios tejidos de rata tratadas con cadmio / Determination of permittivity and conductivity in several rat tissues treated with cadmium

Los campos electromagnéticos ambientales cada vez más presentes en el entorno, pueden afectar a los propios seres humanos y, según su intensidad (magnetoterapia), activar o modular procesos fisiológicos o inducir a posibles patologías. En este trabajo se pretende determinar las características fisicoquímicas (los parámetros permitividad, (e) y conductividad eléctrica (s)) de varios tejidos de rata, mediante la aplicación de microondas de radiofrecuencia, ante variaciones en la concentración de cadmio (Cd) y comprobar si se pueden utilizar como indicadores del efecto específico de la presencia de cadmio, evaluar su efecto por la contaminación del entorno y cuantificar su acúmulo. Por consiguiente, se pretende aportar nuevos datos sobre los efectos de la toxicidad del cadmio en diversos órganos sistémicos y comprobar si existe sinergismo entre el efecto tóxico del cadmio (contaminante ambiental) y el de los campos electromagnéticos. Se utilizan dos grupos de ratas macho Wistar, uno control y otro tratado con Cd (CdCl2 ), con doce dosis (una por día) por vía intraperitoneal desde 0,1 a 1 mg Cd/Kg rata/día. Se extrae la sangre bajo anestesia con éter por punción cardíaca hasta su muerte, y se diseccionan los órganos. Se analizan diversos parámetros sanguíneos, y se congelan los órganos siguientes: hígado, riñones, pulmones, corteza cerebral, testículos, páncreas y músculo, en los que se determina la permitividad y conductividad. Se determinan los coeicientes de reflexión y transmisión de estas muestras a la frecuencia industrial de 2,45 GHz. Esta frecuencia está comprendida dentro del rango de las frecuencias de microondas. Los coeficientes de reflexión y transmisión se miden mediante un analizador de redes, se usa un estimulador HFSS de HP que determina la permitividad de un tejido que produjera los mismos valores de los coeficientes de reflexión y transmisión medidos experimentalmente. Los resultados muestran que en el caso del riñón e hígado los valores de e (permitividad) y s (conductividad) son menores que en el control, lo que se puede explicar porque ambos órganos son los principales tejidos diana de la toxicidad del cadmio. Este elemento provoca disfunción en los túbulos proximales del riñón, y en el caso del hígado, el cadmio también se acumula en este órgano provocando daño hepático, un descenso de la integridad estructural de los hepatocitos y liberación de las transaminasas en el suero. Del análisis de sangre se observa que el cadmio provoca un estado anémico, de forma que se produce una disminución de los valores de hematocrito y hemoglobina con respecto a los controles (AU)

Use of oral anticoagulants in older patients.

Recently published American and British guidelines have comprehensively reviewed the indications for long term anticoagulation. The best evidence currently available supports the use of long term oral anticoagulants in patients with nonvalvular atrial fibrillation (NVAF), venous thromboembolic disease, ischaemic heart disease, mural thrombi, and mechanical heart valves. Selected patients with valvular heart disease, cerebral vascular disease, and peripheral arterial disease may also benefit from the use of these drugs. When no specific contraindications are present, elderly patients with either paroxysmal or persistent NVAF should be considered candidates for treatment with anticoagulants. Pooled analyses of the results from 9 randomised trials demonstrate that warfarin significantly reduces the risk of ischaemic stroke in patients with NVAF, particularly those in a 'high risk' category defined by the presence of additional clinical or echocardiographic risk factors. Long term anticoagulation does not appear to be justified in patients with NVAF considered to be at 'low risk' for stroke. Because the prevalence of NVAF and most other cardiovascular conditions increases with advancing age, many elderly patients will be candidates for thromboprophylaxis. The potential benefit of long term anticoagulation must be carefully weighed against the risk of serious haemorrhage in such patients. Bleeding complications with anticoagulant drugs appear to occur more frequently in older patients than in younger individuals. Advanced age (>75 years), intensity of anticoagulation [International Normalised Ratio (INR) >4.0], history of cerebral vascular disease (recent or remote), and concomitant use of drugs that interfere with haemostasis [aspirin (acetylsalicylic acid) or nonsteroidal anti-inflammatory drugs] are among the most important variables in determining an individual's risk for major bleeding with anticoagulants. Older patients often display increased sensitivity to the effects of warfarin, both in the early induction phase and during the long term maintenance phase of therapy. Conditions such as congestive heart failure, malignancy, malnutrition, diarrhoea and unsuspected vitamin K deficiency, enhance the prothrombin time response. The decision to interrupt anticoagulant therapy before elective surgery in elderly patients should evaluate the thrombotic risk of such a manoeuvre versus the risk of bleeding if anticoagulants are continued. In non-surgical patients, excessively elevated INRs without associated haemorrhage can usually be managed by simply witholding one or several doses of warfarin. If more rapid reversal is needed, small doses of phytomenadione (vitamin K1) can be administered safely without overcorrection or the development of vitamin K-induced warfarin resistance.