Investigation of genetic diversity and polyandry of Leptinotarsa decemlineata using X-linked microsatellite markers.

A panel of X-linked microsatellite markers was newly designed using the data from a previous sequencing project available in NCBI and used for a study of the Colorado potato beetle (CPB, Leptinotarsa decemlineata) X-haplotype variability. The analysis of scaffolds 49 and 61 (newly identified as fragments of CPB chromosome X) found ten high-quality markers, which were arranged in two PCR multiplexes and evaluated in both 420 CPB adults, collected from 14 localities of Czechia and Slovakia, and 866 larvae from five single-female families from two more Czech localities. Length polymorphisms found in 6 loci have predicted 192 potential X-haplotypes, however, only 36 combinations were detected in the adult males (N = 189), and seven additional ones in the larvae. The X-haplotypes were also generally unevenly distributed; five of the most frequent haplotypes were detected in 55% of males, 19 repeating up to ten-times in 38.7% of males and the remained 12 occurred uniquely in 6.3% of males. Bulk analysis of X-haplotypes dissimilarity indicated seven haplotype groups diversified by mutations and recombinations. Two haplotypes showed a distinctive regional distribution, which indicates an east-west disruption of CPB migration probably caused by different environments of localities in the South Bohemia region and Vysocina region. On the contrary, the results indicate a south-north migration corridor alongside the Vltava River. In the single-female families, from 6 to 13 distinct paternal haplotypes were detected, which proved and quantified a frequented polyandry in CPB.

La2Pd2In: superconductivity and lattice properties at ambient and elevated pressures.

Lattice and electronic properties of La2Pd2In were studied at ambient and elevated pressures so as to determine features related to a specific atomic coordination without any influence of magnetism. We describe temperature dependences of lattice parameters, heat capacity and electrical resistivity of single-crystalline La2Pd2In (s.g.P4/mbm) in a broad temperature range 0.09-300 K. Together with the anisotropic effect of hydrostatic pressure, showing that the lattice is more compressible in the basal plane, we can conclude that the lattice is affected by degrees of freedom of the La atoms with positions not imposed by symmetry. The lattice anisotropy is smaller than that found for isostructural ferromagnet Ce2Pd2In. The equilibrium bulk modulusB0= (48 ± 3) GPa was determined on the basis of individual linear compressibilities. Measurement of electrical resistivity indicated a superconducting state belowT= 0.59 K with a low critical field 0.005 T atT= 380 mK. The onset of superconducting state as a bulk property of La2Pd2In was confirmed by measurements of specific heat and AC magnetic susceptibility. Experimental data can be accounted by first-principles electronic-structure calculations based on density-functional theory. The measured Sommerfeld coefficientγ= 10.6 mJ mol-1 K-2, only marginally exceeding the calculatedγ= 9.34 mJ mol-1 K-2, indicates only weak electronic correlations.

Ab initio calculations of the crystal field and phonon dispersions in CePd2Al2 and LaPd2Al2.

CePd2Al2 crystallizes in the CaBe2Ge2-type tetragonal structure (P4/nmm, 129) and undergoes a phase transition to the orthorhombic Cmme structure at around 13 K. Its inelastic neutron spectra reveal an additional magnetic excitation that was ascribed to electron-phonon interaction leading to a formation of a new quantum quasi-bound vibron state. We present the first-principles calculations of the crystal field excitations and lattice dynamics calculations of the phonon dispersions to compare with the experimental data. The calculated crystal field energy splitting in CePd2Al2 agrees well with the model used to describe the experimental neutron scattering spectra. The first excited crystal field level moves to higher energies when undergoing the transformation from tetragonal to orthorhombic structure, in agreement with the experiment. The analysis based on calculated elastic constants and lattice dynamics calculations show that in both tetragonal and orthorhombic structures there are no imaginary modes for any q-wave vector within the Brillouin zone, and therefore the lattice structures are stable. The phonon dispersions and density of states are calculated for both crystal structures of CePd2Al2 and its nonmagnetic counterpart LaPd2Al2. The results generally agree well with the experimental data including the high phonon density of states around 12 meV. The phonon density of states is also used to calculate the mean squared displacement, Debye temperature, lattice heat capacity and compared with similar properties of the available experiment.

Covert postpartum urinary retention.

OBJECTIVES: This study was conducted to determine the frequency of increased postvoiding residual volumes (PVRV) 3 days after delivery and to examine the associated risk factors. BACKGROUND: Increased PVRV ‒ covert postpartum urinary retention, is an asymptomatic condition with possible long-term adverse effects. While early diagnosis and appropriate management can avoid long­term complications, screening is not routinely performed. By identifying risk factors, we could define the group of patients suitable for screening. MATERIAL AND METHODS: This was a prospective observational study carried out over a 3-month period at the university teaching hospital in Bratislava, Slovakia. All participants underwent ultrasound determination of PVRV while 80 ml and more on day 3 was considered pathological. RESULTS: A total of 429 women were included in the study. The prevalence of covert post-partum urinary retention was 9.2 %. Assisted vaginal delivery (ventouse, forceps) and episiotomy were risk factors for post-partum urinary retention (18.7 % vs 6.1 %; p = 0.0053; 52.1 % vs 35.7 %; p = 0.0483; respectively). CONCLUSION: Our observations confirmed the existence of PVRV of 80 ml and more on day 3 in almost 10% of women who had delivered at our clinic. The results of our study prove that instrumental delivery represents a considerable obstetrical-pediatric risk factor for PVRV. Our data support the need of adopting a risk-factor-based approach to PVRV screening as part of postpartum bladder care (Tab. 2, Fig. 1, Ref. 12).

[Renal cell carcinoma with sarcomatoid and rhabdoid features: a clinico-pathological series of 74 cases]. / Carcinomas renales con rasgos sarcomatoides y rabdoides: estudio clínico-patológico de 74 casos.

Objetives. Our aim is to analyze and compare the clinico-pathological features in renal cell carcinomas (RCC) with sarcomatoid and rhaboid phenotype. MATERIAL AND METHODS: We reviewed consecutive patients with nephrectomy RCC from January 1988 to January 2015. The subtyping of the RCC followed the recommendations of the College of American Pathologists. Cases with at least 1% of sarcomatoid and/or rhabdoid change were selected. They were classified as sarcomatoid or rhabdoid according with the predominant morphology, considering the global frecuency of both phenotypes as dedifferentiated component. The following variables were collected: sex, age, symptoms and existence of metastases at diagnosis, parameters listed in the protocol of renal carcinoma of the American College of Pathologists, pattern of tumor growth, perineural invasion, percentage of both tumor necrosis and characteristics of the inflammatory infiltrate. They were described by mean / median or percentage, and compared with Student-t / Mann-Whitney U or ? 2 / Fisher, depending on the sample characteristics. RESULTS: From 1,258 RCC, we identified 45 RCC with sarcomatoid predominance (3,6%) and twenty-nine with rhabdoid predominance (2,3%). RCC with sarcomatoid features showed a higher dedifferentiated component and perineural invasion (27.5 vs. 13.5%, p=0.003 and 28.9 vs. 3.4%, p=0.006, respectively) than RCC with rhabdoid features, while the former showed a higher proportion of neutrophilic inflammation (44.8 vs. 22.2%, p=0.04) and arose more frequently over high grade RCC (55.9 vs. 90.5%, p<0,001). CONCLUSIONS: There was overlapping of the clinico-pathological features of RCC with sarcomatoid and rhaboid phenotype, except for the dedifferentiated component, perineural invasion and neutrophilic inflammation. This close relationship could be explained by a common underlying mechanism, the epithelial-mesenchymal transition, with a double morphological expression that, if confirmed, could lead to selecting patients that would benefit from follow-up or treatment depending on their molecular characteristics.

Carcinomas renales con rasgos sarcomatoides y rabdoides: estudio clínico-patológico de 74 casos / Renal cell carcinoma with sarcomatoid and rhabdoid features: a clinico-pathological series of 74 cases

Fundamento: Nuestro objetivo fue comparar las variables clínico-patológicas de los carcinomas renales (CCR) con fenotipos sarcomatoide y rabdoide. Material y métodos: Se revisaron 1.258 CCR de pacientes consecutivos nefrectomizados entre 1988 y 2015, y se seleccionaron aquellos con ≥1% de cambio sarcomatoide y/o rabdoide. Se clasificaron como sarcomatoide o rabdoide según el fenotipo predominante, considerándose componente desdiferenciado la suma del porcentaje de ambos. Se recopilaron: sexo y edad de los pacientes, síntomas y existencia de metástasis al diagnóstico, parámetros del protocolo de CCR del Colegio Americano de Patólogos, patrón de crecimiento tumoral, invasión perineural, porcentaje de necrosis tumoral y características del infiltrado inflamatorio. Se describieron mediante la media/mediana o el porcentaje y se compararon mediante t de Student/U de Mann-Whitney o χ2/F de Fisher. Resultados: Se identificaron 45 CCR con predominio sarcomatoide (3,6%) y 29 con rabdoide (2,3%); los primeros mostraron mayor componente indiferenciado e invasión perineural respecto a los CCR con rasgos rabdoides (27,5 vs. 13,5%; p=0,003 y 28,9 vs. 3,4%, p=0,006, respectivamente), mientras que estos mostraron doble frecuencia de inflamación neutrofílica (44,8 vs. 22,2%, p=0,04) y surgieron más frecuentemente sobre un CCR de alto grado (55,9 vs. 90,5%, p<0,001). Conclusiones: Los CCR con fenotipos sarcomatoide y rabdoide compartieron características clínico-patológicas, excepto para componente desdiferenciado, invasión perineural, inflamación neutrofílica y origen en CCR de alto grado. Esta similitud sugiere la presencia de un mecanismo común, la transición epitelio-mesénquima, con una expresión morfológica doble que, de confirmarse, podría suponer la posibilidad de seleccionar pacientes para tratamiento o seguimiento a partir de sus características moleculares

Objetives. Our aim is to analyze and compare the clinico-pathological features in renal cell carcinomas (RCC) with sarcomatoidand rhaboid phenotype. Material and methods: We reviewed 1,258 RCC from consecutive patients with nephrectomy from 1988 to 2015, and those with ≥1% of sarcomatoid and/or rhabdoid change were selected. They were classified as sarcomatoid or rhabdoid according with the predominant morphology, considering the global frecuency of both phenotypes as dedifferentiated component. The following variables were collected: sex, age, symptoms and existence of metastases at diagnosis, parameters listed in the protocol of renal carcinoma of the American College of Pathologists, pattern of tumor growth, perineural invasion, percentage of both tumor necrosis and characteristics of the inflammatory infiltrate. They were described by mean/median or percentage, and compared with Student-t/Mann-Whitney U or χ2/Fisher). Results: We identified 45 RCC with sarcomatoid predominance (3,6%) and twenty-nine with rhabdoid predominance (2,3%); the first one showed a higher dedifferentiated component and perineural invasion (27.5 vs. 13.5%, p=0.003 and 28.9 vs. 3.4%, p=0.006, respectively), while the former showed a higher proportion of neutrophilic inflammation (44.8 vs. 22.2%, p=0.04) and arose more frequently over high grade RCC (55.9 vs. 90.5%, p<0,001). Conclusions: There was overlapping of the clinico-pathological features of RCC with sarcomatoid and rhaboid phenotype, except for dedifferentiated component, perineural invasion and neutrophilic inflammation. This close relationship could be explained by a common underlying mechanism, the epithelial-mesenchymal transition, with a double morphological expression that, if confirmed, could lead to selecting patients that would benefit from follow-up or treatment depending on their molecular characteristics

Comparison of shortened and conventional dry period management strategies.

The aim of this study was to compare 2 dry-cow management strategies and evaluate the effect of shortened dry period strategy on feed intake, metabolism, and postpartum performance of dairy cows in early lactation. Twenty-nine high-yielding dairy cows were divided into 2 groups. The control (CON) group (n=14) was assigned to a traditional dry period of approximately 60 d (57±5.9 d) and was fed a far-off dry cow ration from dry-off to -21 d relative to expected parturition. From d -21 relative to expected parturition, the cows were switched to a precalving ration containing an additional 3kg of concentrates. The cows of the experimental group (n=15) were assigned to a shortened dry period (SDP; 35±6.3 d) and were continuously fed a late-lactation diet from d -60 d relative to expected parturition until calving. After calving, both groups were fed the same lactation diet corresponding to their lactation requirements and cows were followed for 100 d of lactation. Prepartum dry matter intake of the cows assigned to an SDP and fed a late-lactation diet was approximately 4.11kg/cow per day greater compared with the CON group during the 60 d. However, no effect of dry period strategy on postpartum dry matter intake was detected. The cows with an SDP produced approximately 2.78kg/d (6.9%) less milk in the first 100 d of lactation than CON cows; the difference was not statistically significant. No differences were observed in live body weight, body condition score, or back-fat thickness between the treatments. Similarly, no differences existed in concentrations of plasma metabolites. The cows of the SDP group showed lower pH and increased concentrations of lactic acid and volatile fatty acids prepartum than the CON cows. Postpartum concentrations of lactic acid, volatile fatty acids, and NH3 and pH in rumen fluid did not differ between the treatments. Shortening of the dry period did not affect the colostrum quality or birth weights of the calves. Based on the results of this study, a traditional dry period management strategy appeared to be more favorable, considering the dry matter intake and milk production, compared with an SDP and feeding a late-lactation diet throughout the dry period.

Physiological and biochemical analysis of overwintering and cold tolerance in two Central European populations of the spruce bark beetle, Ips typographus.

Overwintering success is one of the key aspects affecting the development and outbreaks of the spruce bark beetle, Ips typographus (L.) populations. This paper brings detailed analysis of cold tolerance, and its influence on overwintering success, in two Central European populations of I. typographus during two cold seasons. Evidence for a supercooling strategy in overwintering adults is provided. The lower lethal temperature corresponds well to the supercooling point that ranges between -20 and -22°C during winter months. The supercooled state is stabilized by the absence of internal ice nucleators and by seasonal accumulation of a mixture of sugars and polyols up to the sum concentration of 900 mM. The cryoprotective function of accumulated metabolites is probably based on increasing the osmolality and viscosity of supercooled body fluids and decreasing the relative proportion of water molecules available for lethal formation of ice nuclei. No activity of thermal hysteresis factors (stabilizers of supercooled state) was detected in hemolymph. Lethal times for 50% mortality (Lts50) in the supercooled state at -5, -10 or -15°C are weeks (autumn, spring) or even months (winter), suggesting relatively little mortality caused by chill injury. Lts50 at -15°C are significantly shorter in moist (6.9 days) than in dry (>42 days) microenvironment because there is higher probability of external ice nucleation and occurrence of lethal freezing in the moist situation.

Sentinel lymph node radiolocalization and biopsy in oral cavity and oropharynx mucosal squamous cell carcinoma.

OBJECTIVES: The aims of the study were to assess our feasibility and accuracy of sentinel lymph node radiolocalization in patients with squamous cell carcinoma of the oral cavity and oropharynx, and to determine whether the pathology of the sentinel node reflected regional disease. MATERIAL AND METHODS: Patients preoperatively underwent lymphoscintigraphy after peritumoral injection of a 99m Tc labeled radiocolloid. After perioperative gamma probe radiolocalization of the sentinel lymph nodes, elective neck dissection was performed. The histopathological examination of the sentinel nodes and other nodes of neck dissection specimen were compared. RESULTS: Detection of sentinel lymph nodes by lymphoscintigraphy was feasible in all 12 patients. Also localization with a handheld gamma probe was successful in all patients. Forty sentinel nodes and 276 non-sentinel nodes were histopathologically examined. Occult metastases were confirmed in 7 sentinel nodes (4 patients). There was no false negative sentinel lymph node in our series. CONCLUSION: Identification of the sentinel lymph node in patients with squamous cell carcinoma of the oral cavity and oropharynx is technically feasible and accurate. This method shows to be able to predict occult metastases and select patients who would benefit from neck dissection (Fig. 1, Tab. 2, Ref. 28). Full Text in free PDF www.bmj.sk.

HPLC evaluation of diclofenac in transdermal therapeutic preparations.

High-performance liquid chromatography (HPLC) was selected for analytical evaluation of sodium diclofenac in original transdermal therapeutic preparations containing adjuvant substances (capsaicin, hyoscyamine). After isolation from laminated adhesive patches, diclofenac was analysed on columns with reversed phase, using the mobile phase ethanol and phosphate buffer (pH 6.5) with an addition of tetrabutylammonium iodide and detection at 284 nm. Not only the total amount of diclofenac in the patch was evaluated, but HPLC methodology was also employed to select a suitable acceptor medium for permeation experiments. In patches manufactured in the tested series, HPLC was also employed to examine the release of diclofenac and its in vitro permeation through the human skin.

Characterization of a candidate Trypanosoma brucei U1 small nuclear RNA gene.

We have previously shown that the poly(A) polymerase (PAP) gene of Trypanosoma brucei is interrupted by an intervening sequence. It was postulated that removing this intron by cis-splicing requires a yet unidentified U1 small nuclear RNA (snRNA), which in other organisms engages in base-pair interactions across the 5' splice site during early spliceosome assembly. Here we present a characterization of a 75 nucleotide long candidate T. brucei U1 snRNA. Immunoprecipitation studies indicate that a trimethylguanosine cap structure is present at the 5' end and that the RNA is bound to core proteins common to spliceosomal ribonucleoprotein particles. The U1 snRNA has the potential for extensive intermolecular base pairing with the PAP 5' splice site. We used block replacement mutagenesis to identify sequences necessary for in vivo expression of U1 snRNA. We found that at least two cis-acting elements, tRNA-like A and B boxes, located in the 5'-flanking region are necessary for U1 snRNA synthesis; no internal sequences close to the transcription start site are essential, suggesting a promoter architecture distinct from other trypanosome U-snRNA genes.

Esters of 6-dimethylaminohexanoic acid as skin penetration enhancers.

We prepared a series of five esters of 6-dimethylaminohexanoic acid, and characterised the compounds by their NMR and IR spectra. Their ability to function as transdermal penetration enhancers was subsequently evaluated using excised human skin as a membrane, modified Franz diffusion cells, and theophylline as a model permeant. The penetration-enhancing efficiency of esters 1-5 was studied in the donor media of propylene glycol and isopropyl myristate, and expressed as the corresponding enhancement factors (EF). All the esters increased the penetration of theophylline through the skin. The enhancement factor for the most active substance, undecyl 6-dimethylaminohexanoate, was 118.5 (+/- 19) from propylene glycol.

HPLC evaluation of drugs in transdermal therapeutic preparations. I. Glycerol trinitrate.

In connection with the development of transdermal delivery systems (TTS) of the laminated adhesive patches type, HPLC was selected for analytical evaluation of active principles. It was employed for analysis of glycerol trinitrate (GTN) as one of the drugs administered in the form of medicinal adhesive patches. After isolation from the patch, GTN was analysed by reverse phase HPLC, employing methanol and water as the mobile phase, and detected at 206 nm. The total amount of GTN in the patch was evaluated and its concentrations in a lactose trituration and the reservoir layer were determined. In the patches manufactured in a test series, the elaborated HPLC method was used to investigate the release of GTN in a liberation study and its permeation through excised human skin in experiments performed in vitro. In connection with the stability study, the decomposition products of GTN were also determined.

Crude protein degradability, protein digestible in the intestine and net energy for lactation of whole crop barley in various vegetative stages.

Crude protein degradability (CP deg), protein digestible in the intestine (PDI) and net energy of lactation (NEL) content of whole crop spring barley were estimated by means of the in situ method using three steers fitted with rumen cannulas and fed near the maintenance level of nutrition. Barley was cut in five consecutive vegetative stages (end of the earing, S1, end of the flowering, S2, middle of the milky stage of the grain, S3, beginning of the dough stage, S4, middle of the dough stage, S5, including S5 silage). CP deg, PDI and NEL were significantly (P < 0.01) affected by the growth stage. The relationships of the CP deg (%), PDI (g/kg DM) and NEL (MJ/kg DM) to the vegetative stage of barley quantified on a decimal scale (X) were as follows: CP deg = 40.1 + 0.46X (I2yx = 82%), PDI = -330.8 + 12.86X-0.1007 X2 (I2yx = 83%), and NEL = -0.48 + 0.0802X (I2yx = 86%), resp. Compared with drying at 60 degrees C ensiling increased CP deg (P < 0.01) and NEL (P < 0.05), whereas PDI was not affected (P > 0.05).

Esters of omega-amino acids as flexible penetration enhancers.

Series of esters of 6-aminohexanoic acid, 7-aminoheptanoic acid, and 8-aminooctanoic acid were synthesized as candidates for novel skin penetration enhancers. The permeation of theophylline expressed in the form of the enhancement factor (EF) was studied with human cadaver skin by a diffusion cell technique in the presence of the esters 1-17 in the water vehicle. The permeation in the presence of the 6-aminohexanoates 1-5 was also studied in the olive oil vehicle. Of the compounds tested, the most satisfactory enhancement in the water vehicle and oil vehicle were observed with octyl 6-aminohexanoate (EF = 45.5) and decyl 6-aminohexanoate (EF = 19.5), respectively.

epsilon-Aminocaproic acid esters as transdermal penetration enhancing agents.

The synthesis of epsilon-aminocaproic acid esters is described. Two representative members from a group of five of the 1-alkyl homologues synthetized as flexible analogues of 1-alkylazacyclohepatanone derivatives were evaluated in vitro for their effectiveness on the transport of theophylline through the excised human cadaver skin in comparison with Azone. The 1-octyl- and 1-dodecyl-epsilon-aminocaproic acid esters (OCEAC and DDEAC) show excellent penetration enhancement. Donor samples contained 2.5% theophylline and 1% enhancers tested in three different vehicles. Fluxes of theophylline were increased with OCEAC about 19 times from olive oil, 45 times from water, and about 38 times from water-propylene glycol (3:2) vehicle toward controls (with DDEAC about 17, 39, and 35 times, respectively) and they were markedly higher than Azone under the given conditions. Acute LD50's (i.p. in mice) of OCEAC (DDEAC) were 245 mg/kg (352 mg/kg), with a slightly lower toxicity than Azone. OCEAC and DDEAC did not exhibit acute dermal irritation in vivo on rabbits at a 5% concentration in white petrolatum.

[Interactions of drugs with PHEMA hydrogel]. / Interakce lécivých látek s PHEMA hydrogelem.

Hydration of polyhydroxyethylmethacrylate (PHEMAs hydrogel with aqueous solutions of medicinal substance) renders it possible to deposit medicinal substances within the hydrogel. At the same time the concentration of the medicinal substances in the hydrogel is higher than their concentration in the hydrating solution. For the nonspecific interactions, which affect the deposition of medicinal substances in hydrogel, their surface tension is of principal importance. With decreasing surface tension the amount of medicinal substances in hydrogel is increased and simultaneously their release into the aqueous medium is decelerated. Interactions of six selected medicinal substances with PHEMA hydrogel were experimentally checked in the in vitro conditions.