Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.

Red blood cell (RBC) parameters such as morphology, volume, refractive index, and hemoglobin content are of great importance for diagnostic purposes. Existing approaches require complicated calibration procedures and robust cell perturbation. As a result, reference values for normal RBC differ depending on the method used. We present a way for measuring parameters of intact individual RBCs by using digital holographic microscopy (DHM), a new interferometric and label-free technique with nanometric axial sensitivity. The results are compared with values achieved by conventional techniques for RBC of the same donor and previously published figures. A DHM equipped with a laser diode (lambda = 663 nm) was used to record holograms in an off-axis geometry. Measurements of both RBC refractive indices and volumes were achieved via monitoring the quantitative phase map of RBC by means of a sequential perfusion of two isotonic solutions with different refractive indices obtained by the use of Nycodenz (decoupling procedure). Volume of RBCs labeled by membrane dye Dil was analyzed by confocal microscopy. The mean cell volume (MCV), red blood cell distribution width (RDW), and mean cell hemoglobin concentration (MCHC) were also measured with an impedance volume analyzer. DHM yielded RBC refractive index n = 1.418 +/- 0.012, volume 83 +/- 14 fl, MCH = 29.9 pg, and MCHC 362 +/- 40 g/l. Erythrocyte MCV, MCH, and MCHC achieved by an impedance volume analyzer were 82 fl, 28.6 pg, and 349 g/l, respectively. Confocal microscopy yielded 91 +/- 17 fl for RBC volume. In conclusion, DHM in combination with a decoupling procedure allows measuring noninvasively volume, refractive index, and hemoglobin content of single-living RBCs with a high accuracy.

Comparison of the reproductive behavior between isolated Peckia chrysostoma (Wiedemann, 1830) and Adiscochaeta ingens (Walker, 1849) (Diptera: Sarcophagidae) females reared in laboratory

In both species, maintained under laboratory environmental conditions, anautogeny was comproved and all females that had free access to proteic source were fertiles. We obtained the following average values for Peckiachrysostoma: 59.7 ± 15.6 and 81.8 ± 15.4 days of longevity in the respective cases of free access and no access to proteic source, 21.4 ± 4.3 days of pre-larviposition period and 35.2 ± 16.5 days of larviposition period, 5.3 ± 1.8 larvipositions female with 7.0 ± 1.1 days of periodicity, 35.7 ± 6.1 larvae per larviposition leading to a total number of 183.8 ± 69.2 viable larvae per female and 94.8% ± 5.3% of productivity. The mean number of ovarioles per female was 56.4 ± 9.8, resulting in a reproductive potential of 63.3%. For Adiscochaeta ingens, the obtained average values were: 41.3 ± 6.3 and 52 ± 13.1 days of longevity in the respective cases of free access and no access to proteic source, 15.3 ± 1.7 days of pre-larviposition period and 21.5 ± 7.5 days of larviposition period, 3 ± 0.7 larvipositions per female with 10.4 ± 0.8 days of periodicity, 30.3 ± 8.2 larvae per larviposition leading to a total number of 78.5 ± 21.7 viable larvae per female and 90.1% ± 16% of productivity. The mean number of ovarioles per female was 54.6 ± 5.2, resulting in a reproductive potential of 55.5%. Within applied parameters, the values obtained for P. chrysostoma demonstrate its superior productivity in comparison with A. ingens

A vertebrate reproductive system involving three ploidy levels : hybrid origin of triploids in a contact zone of diploid and tetraploid green toads (Bufo viridis subgroup)

The rise and consequences of polyploidy in vertebrates, whose origin was associated with genome duplications, may be best studied in natural diploid and polyploid populations. In a diploid/tetraploid (2n/4n) geographic contact zone of Palearctic green toads in northern Kyrgyzstan, we examine 4ns and triploids (3n) of unknown genetic composition and origins. Using mitochondrial and nuclear sequence, and nuclear microsatellite markers in 84 individuals, we show that 4n (Bufo pewzowi) are allopolyploids, with a geographically proximate 2n species (B. turanensis) being their maternal ancestor and their paternal ancestor as yet unidentified. Local 3n forms arise through hybridization. Adult 3n mature males (B. turanensis mtDNA) have 2n mothers and 4n fathers, but seem distinguishable by nuclear profiles from partly aneuploid 3n tadpoles (with B. pewzowi mtDNA). These observations suggest multiple pathways to the formation of triploids in the contact zone, involving both reciprocal origins. To explain the phenomena in the system, we favor a hypothesis where 3n males (with B. turanensis mtDNA) backcross with 4n and 2n females. Together with previous studies of a separately evolved, sexually reproducing 3n lineage, these observations reveal complex reproductive interactions among toads of different ploidy levels and multiple pathways to the evolution of polyploid lineages.

The neuropeptide Y Y1 receptor regulates leptin-mediated control of energy homeostasis and reproductive functions.

The orexigenic neurotransmitter neuropeptide Y (NPY) plays a central role in the hypothalamic control of food intake and energy balance. NPY also exerts an inhibition of the gonadotrope axis that could be important in the response to poor metabolic conditions. In contrast, leptin provides an anorexigenic signal to centrally control the body needs in energy. Moreover, leptin contributes to preserve adequate reproductive functions by stimulating the activity of the gonadotrope axis. It is of interest that hypothalamic NPY represents a primary target of leptin actions. To evaluate the importance of the NPY Y1 and Y5 receptors in the downstream pathways modulated by leptin and controlling energy metabolism as well as the activity of the gonadotrope axis, we studied the effects of leptin administration on food intake and reproductive functions in mice deficient for the expression of either the Y1 or the Y5 receptor. Furthermore, the role of the Y1 receptor in leptin resistance was determined in leptin-deficient ob/ob mice bearing a null mutation in the NPY Y1 locus. Results point to a crucial role for the NPY Y1 receptor in mediating the NPY pathways situated downstream of leptin actions and controlling food intake, the onset of puberty, and the maintenance of reproductive functions.

The reproductive capacity of Panstrongylus megistus (Hemiptera: Reduviidae) males treated with precocenes

Reproductive performance including mating, semen transfer, egg production and hatching rate were investigated in pairs of Panstrongylus megistus in wich the male was treated with either precocene II or ethoxyprecocene II, topically applied shortly after imaginal molt. Eighty percent of the males were not able to mate for a period as long as 40 days after treatment. Those males wich did mate (20%) produced small spermatophores; a few (10%) were unable to transfer spermatozoa to female, while the others (10%) exhibited reproductive performance similar to that of non-treated males. The cumulative effects of the precocene on the different stages of reproduction resulted in a drastic reduction of offspring production. The anti-allatal compound precocene seems to inhibit successful reproduction through a corpus allatum inactivation and subsequent lack of secretory activity of the male accessory glands.

Indications et techniques actuelles de reconstruction mammaire après mastectomie [Indications and techniques of reconstruction after mastectomy].

Reconstructive surgery takes an important place in breast cancer treatment. Immediate breast reconstruction is performed during the same operation as mastectomy. It is contraindicated following radiotherapy. Reconstruction performed after mastectomy is called differed breast reconstruction. It is completed 6 months after chemotherapy and 1 year after radiotherapy. Prosthetic breast reconstruction is indicated when tissues are of good qualities and breast are small. Autologous reconstruction is performed in case of radiotherapy or large breast. After breast reconstruction, imperfections can be corrected with autologous fat injection.

High magnification cystoscopy in the primary diagnosis of bladder tumors.

Purpose of reviewIn bladder cancer, discrimination between benign and malignant tissue may remain tricky with current endoscopic tools. On the basis of our recent experience with high-magnification cystoscopy, compared with other tools such as optical coherence tomography or confocal laser endomicroscopy, it is suggested here that this discrimination may well be feasible endoscopically. The clinical potential of these systems that are being developed as complementary tools to the current endoscopic equipment is reviewed.Recent findingsAt present, white-light cystoscopy, either assisted by fluorescence cystoscopy or narrow-band imaging, is proposed for the global cystoscopic examination of bladder cancer patients. Both techniques compete to help to reduce the recurrence rate by improving exophytic tumor detection, and the extent of carcinoma in situ and high-grade dysplasia. All of which are important prognosis factors for disease progression. In addition, recent findings on neoangiogenesis that accompanies early stage bladder cancer show that this may also be an important observable switch in bladder cancerogenesis, as it is found very early in tumor development. The high magnification cystoscopy as a complementary tool to fluorescence cystoscopy allows classification of the vessel patterns on fluorescence positive sites, and thus facilitates the discrimination between cancerous and noncancerous lesions. This information may be useful to reduce the false positive rate of fluorescence cystoscopy.SummaryEmerging technologies aiming at a real-time in-situ discrimination between benign and malignant tissue during endoscopic bladder exploration is a promising development for the monitoring of bladder cancer patients.

Effects of gamma irradiation on the reproductive capacity and the longevity of Panstrongylus (Hemiptera: Reduviidae)

The effects of radiation on the reproductive capacity and the longevity of Panstrongylus magistus were studied. An indirect correlation between longevity and radiation doses was observed. Males were more affected than females. Longevity of females submitted to 10 Gy was not different when compared to controls. Some of the irradiated males copulated and in these cases semen was transferred to females, but only few eggs were laid. Fertility was seriously affected in all irradiated groups, decreasing with increasing intensity of dose. The dose of 80 Gy induced sterility in males and females. Gelatinous spermatophores that were expelled by females irradiated with 20 and 40 Gy, may be a consequence of irradiation that induced modification in the bursa copulatrix pH.

Techniques for analytical testing of unconventional samples.

Forensic scientists have long detected the presence of drugs and their metabolites in biological materials using body fluids such as urine, blood and/or other biological liquids or tissues. For doping analysis, only urine has so far been collected. In recent years, remarkable advances in sensitive analytical techniques have encouraged the analysis of drugs in unconventional biological samples such as hair, saliva and sweat. These samples are easily collected, although drug levels are often lower than the corresponding levels in urine or blood. This chapter reviews recent studies in the detection of doping agents in hair, saliva and sweat. Sampling, analytical procedures and interpretation of the results are discussed in comparison with those obtained from urine and blood samples.

Reliable acetabular cup orientation with a new gravity-assisted guidance system.

Acetabular cup orientation is a key factor determining hip stability, and standard mechanical guides have shown little help in improving alignment. An in vitro study was carried out to compare the accuracy and precision of a new gravity-assisted guidance system with a standard mechanical guide. Three hundred ten cups were impacted by 5 surgeons, and the final cup orientation was measured. With the new guide, the average error in anteversion was 0.4 degrees , compared with 10.4 degrees with the standard guide and 0.3 degrees and -4.7 degrees , respectively, for abduction angles. The average time required for orienting the cups was similar for both guides. The accuracy and reproducibility obtained with the new guide were better (P < .0001). These good results would require a clinical validation.

Protein recovery, separation and purification: selection of optimal techniques using an expert system

The paper discusses the utilization of new techniques ot select processes for protein recovery, separation and purification. It describesa rational approach that uses fundamental databases of proteins molecules to simplify the complex problem of choosing high resolution separation methods for multi component mixtures. It examines the role of modern computer techniques to help solving these questions.

Estudi de l'organogènesi del sistema reproductor de la cabra de mar (Maja brachydactyla) i efecte dels factors ambientals sobre la reproducció en captivitat

Diferents aspectes de la reproducció de la cabra de mar Maja brachydactyla s’han estudiat amb l’objectiu d’obtenir coneixements bàsics i aplicats al cultiu en captivitat d’aquesta espècie de gran interès comercial i aqüícola. L’anatomia interna de l’aparell reproductor masculí, el procés d’espermatogènesi, amb especial èmfasi amb la formació de l’acrosoma, i l’estructura i organització l’espermatozoide s’han descrit amb tècniques microscòpiques avançades. El desenvolupament de les gònades durant els primeres fases del desenvolupament post- embrionari (fases larvàries i primer cranc juvenil) han estat descrites mitjançant la quantificació de l’expressió del gen vasa, el qual és expressat específicament per les cèl•lules de la línia germinal. Aquests coneixements són bàsics per a la comprensió del paper dels mascles en la la reproducció i la seva aplicació posterior en condicions de captivitat. La reproducció en captivitat s’ha estudiat amb una sèrie d’experiments amb l’objectiu de determinar la quantitat, pes sec i composició bioquímica proximal de les larves acabades d’eclosionar en condicions de captivitat i l’efecte de les condicions ambientals, com ara el fotoperíode i la salinitat, sobre aquests paràmetres. Aquests experiments demostren què es poden obtenir larves de la cabra de mar en condicions de captivitat què podrien ser utilitzades per a la producció i l’efecte sobre la reproducció del fotoperíode i salinitat.

Effects of three organophosphorus insecticides in the reproductive potential of Culex quinquefasciatus

A Culex quinquefasciatus Say 1823 strain with resistant genes to organophosphates was tested in the laboratory to know the reproductive potential after exposure, as larvae, at the LC30 and LC70 (mg/l) of three organophosphorus insecticides: malathion, chlorpyrifos and methyl-pirimiphos. Data showed that fecundity was decreased significantly by malathion at LC30 = 0.0025 and LC70 = 0.0075, whereas fertility has a no significant decrement by chlorpyrifos and methyl-pirimiphos at the LC70 (0.000016, 0.00043). The sexual index was affected by chlorpyrifos and methyl-pirimiphos showing a greater number of adult females.