Tenrec phylogeny and the noninvasive extraction of nuclear DNA

Due in part to scarcity of material, no published study has yet cladistically addressed the systematics of living and fossil Tenrecidae (Mammalia, Afrotheria). Using a noninvasive technique for sampling nuclear DNA from museum specimens, we investigate the evolution of the Tenrecidae and assess the extent to which tenrecids fit patterns of relationships proposed for other terrestrial mammals on Madagascar. Application of several tree-reconstruction techniques on sequences of the nuclear growth hormone receptor gene and morphological data for all recognized tenrecid genera supports monophyly of Malagasy tenrecids to the exclusion of the two living African genera. However, both parsimony and Bayesian methods favor a close relationship between fossil African tenrecs and the Malagasy Geogale, supporting the hypothesis of island paraphyly, but not polyphyly. More generally, the noninvasive extraction technique can be applied with minimal risk to rare/unique specimens and, by better utilizing museum collections for genetic work, can greatly mitigate field expenses and disturbance of natural populations.     

Analysis of noninvasive macromolecular transport measurements in the lung

Several groups of investigators are measuring transcapillary protein flux in the lung using noninvasive methods. Results from these studies are reported using several different protein transport indexes, including pulmonary transvascular transfer coefficient, relative extravascular protein, pulmonary transcapillary escape rate, protein leak index, lung transferrin index, slope index, and lung-to-heart count ratios. The purpose of this study is to discover the relationships between these indexes by employing a two-compartment theory of protein transcapillary transport in the lung. We found that all the above indexes can be related to a single index, which we call the normalized slope index. This index is the time rate of change of radioactivity originating from protein in lung interstitium divided by radioactivity arising from protein in lung plasma, normalized by this ratio at time 0, and corrected for blood volume changes. In particular the normalized slope index is shown to be the same as pulmonary transcapillary escape rate under normal sampling conditions and is relatively unaffected by changes in interstitial volume. The response of the normalized slope index to changes in microvascular pressure and microvascular permeability is explored by applying a two-pore model of the microvascular barrier. Results indicate that the normalized slope index is relatively insensitive to changes in microvascular pressure but is greatly affected by changes in microvascular permeability (i.e., changes in large-pore size or number). Since all published leak indexes are related, we would encourage all investigators in the field to adopt a single leak index. We recommend that when a two-compartment model is applied to external detection data, the results be expressed as pulmonary transcapillary escape rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cytologic study of noninvasive intraductal papillary-mucinous carcinoma of the pancreas

OBJECTIVE: To examine the cytologic features of noninvasive intraductal papillary-mucinous carcinoma (IPMC) in an attempt to differentiate it cytologically from hyperplasia, invasive IPMC and invasive ductal adenocarcinoma (IDA). STUDY DESIGN: Tumor samples from 23 patients clinically diagnosed with a mucin-producing tumor of the pancreas, including 10 cases of hyperplasia, 10 noninvasive IPMCs and 3 invasive IPMCs, and tumor samples from 21 patients with IDA, were examined cytologically. Cytologic specimens were obtained in various ways, such as by fine needle aspiration, imprint, brushing, vinyl tube aspiration and aspiration from a cannula at endoscopic retrograde cholangiopancreatography. RESULTS: Cytologically, all 10 noninvasive IPMCs exhibited clearly defined cytoplasmic boundaries (+1, ++4, ++ and mainly small nuclei (++7, +++3), with nuclear size and cytoplasm similar to those of hyperplastic cells, and irregular chromatin distribution (+2, ++8) and prominent nucleoli (-2, +4, ++3, +++1), all atypical nuclear features similar to those of invasive IPMC or IDA. There also were small papillary cohesive clusters (-1, +2, ++4, +++3) and euchromatin (+1, ++1, +++8), which were found only in IPMCs. CONCLUSION: The cytologic features of small, malignant nuclei and small papillary cohesive clusters are suggestive of noninvasive IPMC. Furthermore, the addition of clearly defined cell borders and euchromatin and the presence of some goblet cells are more strongly suggestive of noninvasive IPMC.     

Current complex noninvasive diagnosis of parathyroid tumors

The diagnosis of bulky formations of the parathyroid glands (PTG) has become possible since current high-resolution techniques of visualization, such as ultrasound study (USS), computed tomography (CT), magnetic resonance imaging (MRI), were introduced into practice. The presence of clinical and/or laboratory signs of hyperparathyroidism (HPT) is a signal to initiate a goal-oriented search for abnormal PTG formations. The complex diagnosis of HPT involves the methods of detecting osteoporosis ranging from routine X-ray study of the hand and foot to more in-depth techniques: dichromatic X-ray absorptiometry (DXA) and quantitative CT (QCT). USS is an excellent method for screening if abnormal PTC changes are suspected; however, negative USS results in the presence of clinical and/or laboratory signs of HPT should not stop a diagnostic search. CT with intravenous contrast bolus specifies the site and structure of an formation, has some advantage in detecting retrosternal tumors. Due to its high tissue contrast, three-dimensional images, none ionizing radiation and osseous structural artefacts, MRI becomes a preferable tool for studying PTG when they are typically or atypically located. Needle biopsy is required when noninvasive methods cannot characterize the pattern of an abnormal PTG formation properly or their results are contradictory.     

Quantifying genotyping errors in noninvasive population genetics

The use of noninvasively collected samples greatly expands the range of ecological issues that may be investigated through population genetics. Furthermore, the difficulty of obtaining reliable genotypes with samples containing low quantities of amplifiable DNA may be overcome by designing optimal genotyping schemes. Such protocols are mainly determined by the rates of genotyping errors caused by false alleles and allelic dropouts. These errors may not be avoided through laboratory procedure and hence must be quantified. However, the definition of genotyping error rates remains elusive and various estimation methods have been reported in the literature. In this paper we proposed accurate codification for the frequencies of false alleles and allelic dropouts. We then reviewed other estimation methods employed in hair- or faeces-based population genetics studies and modelled the bias associated with erroneous methods. It is emphasized that error rates may be substantially underestimated when using an erroneous approach. Genotyping error rates may be important determinants of the outcome of noninvasive studies and hence should be carefully computed and reported.     

Numerical simulation of noninvasive blood pressure measurement

In this paper, a simulation model based on the partially pressurized collapsible tube model for reproducing noninvasive blood pressure measurement is presented. The model consists of a collapsible tube, which models the pressurized part of the artery, rigid pipes connected to the collapsible tube, which model proximal and distal region far from the pressurized part, and the Windkessel model, which represents the capacitance and the resistance of the distal part of the circulation. The blood flow is simplified to a one-dimensional system. Collapse and expansion of the tube is represented by the change in the cross-sectional area of the tube considering the force balance acting on the tube membrane in the direction normal to the tube axis. They are solved using the Runge-Kutta method. This simple model can easily reproduce the oscillation of inner fluid and corresponding tube collapse typical for the Korotkoff sounds generated by the cuff pressure. The numerical result is compared with the experiment and shows good agreement.     

Study of the distribution of mustelids over the Southern Urals using noninvasive methods

Noninvasive methods (interviews, feces collection, DNA analysis, and trail camera traps) allowed us to identify the most widespread mustelid species and assess the pattern of their distribution over the territory of the region. Two mustelid species—the American mink and common marten—are predominant in the Southern Urals. In summer and autumn, the American mink prefers brooks emptying into large rivers, whereas the common marten is more frequently met along banks of forest brooks and rivers with channel widths not exceeding 3 m. Whereas in usual conditions we observe spatial segregation between different mustelid species, extreme situations force mustelids to form multispecies communities on the banks of water bodies, where a mutual avoidance is reached by the temporal segregation strategy of circadian activity.     

A theoretical model for the noninvasive assessment of the transmitral pressure-flow relation.

The purpose of this paper is to formulate from the equations of fluid mechanics an equation which describes the transmitral pressure-flow relationship. According to the linear momentum equation applied to the atrioventricular coupling, the left-atrium-left-ventricle pressure difference (Pa-Pv) can be written as Pa-P v = A delta v/delta t + B v 2 + C v, where v is the transmitral blood velocity and A, B, and C are variables related to the geometry of the atrium, ventricle and mitral orifice, respectively. Based on this theory, Pa-Pv is calculated noninvasively in a patient with a nonobstructive mitral valve. Mitral flow and cardiac dimensions recorded by Doppler echocardiography are digitized and analyzed. Calculation shows that Pa-Pv reaches its peak value at the time of flow peak acceleration and has already considerably decreased at the time of peak velocity. The time course of calculated Pa-Pv is in close agreement with the published experimental catherization data. Numerical computation of early diastolic left atrium and left ventricle pressure curves based on the experimental data of others for the time constant of left ventricular relaxation, left atrial and ventricular chambers stiffness constants, combined with sine-waveform-simulated mitral flow, verifies the time course and the magnitude of Pa-Pv as predicted from flow equations. This paper provides a theoretical method for the noninvasive assessment of the transmitral pressure-flow relationship using ultrasound technique and might help to achieve a better understanding of the diastolic function as assessed by Doppler echocardiography.     

Nucleus transfer in mammals: noninvasive approaches for the preparation of cytoplasts

The production of cloned animals is a difficult and complex procedure that requires two basic steps. First, the cytoplast must be prepared by the enucleation of metaphase II oocytes. Second, the nucleus is transferred either by fusion or by direct microinjection into the cytoplast. The preparation of cytoplasts is a crucial step because they must be able to reprogram the transferred nucleus and to secure the development of reconstructed embryos. Moreover, the classical mechanical enucleation of metaphase II oocytes is rather technically difficult, requiring good equipment and considerable micromanipulation skill. For this reason the simplification of this step is permanently in the centre of interest of those scientists who are involved in the production of clones.     

A noninvasive intraoral electromyographic electrode for genioglossus muscle

We have developed an intraoral bipolar surface electrode for the genioglossus muscle. The electrode, made from an athletic mouthguard and dental impression material, was fitted to the lower teeth. Electrode wires, bared at the tip, were positioned on the bottom of the mouthpiece to lie in contact with the superior surface of the genioglossus just behind the teeth. The electromyographic activity of the genioglossus, simultaneously obtained from the surface electrode and conventional intramuscular electrodes, was compared during quiet breathing, CO2 rebreathing, and a variety of tongue movements. The two types of electrodes recorded similar patterns of muscle activity, and spectral analyses of the signals revealed similar and highly coherent frequency spectra. We conclude that the surface electrode satisfactorily reflects the bioelectrical activity of the genioglossus. The mouthpiece electrode has the further advantage that quantitative comparisons can be made among recordings made in different experimental sessions, since the fit of the mouthpiece to the teeth assures a constant relationship of the electrode to the genioglossus muscle.