Oxidative stress in dairy cows naturally infected with the lungworm Dictyocaulus viviparus (Nematoda: Trichostrongyloidea).

The aim of this study was to analyse the oxidative and anti-oxidant status in serum samples from dairy cows naturally infected by Dictyocaulus viviparus and its relation with pathological analyses. The diagnosis of the disease was confirmed by necropsy of one dairy cow with heavy infection by the parasite in the lungs and bronchi. Later, blood and faeces were collected from another 22 cows from the same farm to measure reactive oxygen species (ROS) levels, thiobarbituric acid-reactive substances (TBARS), catalase (CAT) and superoxide dismutase (SOD) activities on day 0 (pre-treatment) and day 10 (post-treatment with eprinomectin). Faecal examination confirmed the infection in all lactating cows. However, the number of D. viviparus larvae per gram of faeces varied between animals. Cows showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal secretion). Further, they were classified and divided into two groups: those with mild (n = 10) and severe disease (n = 12). Increased levels of TBARS (P < 0.001), ROS (P = 0.002) and SOD activity (P < 0.001), as well as reduced CAT activity (P < 0.001) were observed in cows with severe clinical signs of the disease compared to those with mild clinical signs. Eprinomectin treatment (day 10) caused a reduction of ROS levels (P = 0.006) and SOD activity (P < 0.001), and an increase of CAT activity (P = 0.05) compared to day 0 (pre-treatment). TBARS levels did not differ with treatment (P = 0.11). In summary, increased ROS production and lipid peroxidation altered CAT and SOD activities, as an adaptive response against D. viviparus infection, contributing to the occurrence of oxidative stress and severity of the disease. Treatment with eprinomectin eliminated the infection, and thus minimized oxidative stress in dairy cows.

Correlation versus hybridization gap in CaMn[Formula: see text]Bi[Formula: see text].

We study the interplay between electronic correlations and hybridization in the low-energy electronic structure of CaMn[Formula: see text]Bi[Formula: see text], a candidate hybridization-gap semiconductor. By employing a DFT+U approach we find both the antiferromagnetic Néel order and band gap in good agreement with the corresponding experimental values. Under hydrostatic pressure, we find a crossover from hybridization gap to charge-transfer insulting physics due to the delicate balance of hybridization and correlations. Increasing the pressure above [Formula: see text] GPa we find a simultaneous pressure-induced volume collapse, plane-to-chain, insulator to metal transition. Finally, we have also analyzed the topology in the antiferromagnetic CaMn[Formula: see text]Bi[Formula: see text] for all pressures studied.

In high risk hypertensive subjects with incidental and unilateral renal artery stenosis percutaneous revascularization with stent improves blood pressure control but not glomerular filtration rate.

AIM: In high-risk hypertensive subjects (HTs) with incidental unilateral renal artery stenosis (RAS), the effectiveness of percutaneous revascularization with stent (PR-STENT) on blood pressure (BP) and glomerular filtration rate (GFR) is not established. METHODS: Eighteen HTs aged 65.7 ± 9.2 years with angiographically diagnosed unilateral RAS (≥ 60%) were randomized to receive PR-STENT (N=9) or to NO-STENT (N=9). BP (mercury sphygmomanometer) and GFR (99mTc-DTPA clearances during renal scintigraphy) were evaluated yearly for three years. Echo-Doppler of renal arteries was performed to verify the anatomic patency and flow velocities of the reperfused artery. Analysis of variance compared BP and GFR values changes from baseline to the follow-up; differences for continuous variables were evaluated between groups with the Tukey's post hoc test after adjustment for age, change of BP between baseline and at the follow-up, GFR and body mass index (BMI). RESULTS: Baseline systolic BP and GFR values were not different between groups. The significantly greater GFR increase observed in PR-STENT than in NO-STENT at univariate analysis at the end of follow-up (62.5 ± 19.2 vs. 42.24 ± 17.6, P<0.02) disappeared after adjustment for confounding factors. However, systolic BP remained significantly lower in PR-STENT than in NO-STENT (140.1 ± 4.6 vs. 170.0 ± 8.3, P<0.0001) also after adjustment for age, GFR and BMI. CONCLUSION: PR-STENT reduces systolic BP without improving GFR. Due to the strong association between high BP and renal damage, this study raises the question on whether PR-STENT should be performed in all HTs with unilateral and incidental RAS.

Dermoscopy as a Tool for Estimating Breslow Thickness in Melanoma. / La dermatoscopia como herramienta para inferir el Breslow del melanoma.

BACKGROUND AND OBJECTIVE: The incidence of melanoma has increased significantly, and early diagnosis is the most effective way to reduce associated deaths. Dermoscopy increases diagnostic accuracy in melanoma and analysis of dermoscopic structures can help in the estimation of tumor thickness. The aim of this study was to analyze the influence of Breslow thickness on the dermoscopic characteristics of melanoma. MATERIAL AND METHODS: Observational, cross-sectional study of patients with histologically confirmed melanoma and dermoscopic images of the tumor. The patients were divided into three groups: melanoma in situ, thin melanoma (≥ 1 mm Breslow thickness), and thick melanoma (≥ 1 mm Breslow thickness). Age, sex, tumor location, and histologic and dermoscopic characteristics were analyzed in all cases. RESULTS: We studied 215 patients: 88 with melanoma in situ, 73 with thin melanoma, and 54 with thick melanoma. The frequency of the following dermoscopic features increased with increasing Breslow thickness: the blue-white veil (p < 0.001), white shiny structures (p < 0.001), and milky-red areas (p < 0.003). Angulated lines, by contrast, became less common with increasing thickness (p < 0.002). CONCLUSIONS: Dermoscopy not only improves diagnostic accuracy for pigmented lesions but also helps in the preoperative assessment of Breslow thickness in melanoma.

Immunization with the recombinant myosin regulatory light chain (FhrMRLC) in Adjuplex® adjuvant elicits a Th1-biased immune response and a reduction of parasite burden in Fasciola hepatica infected rats.

The aim of this study was to assess the immune response and the protective efficacy elicited by the vaccination with the recombinant Fasciola hepatica myosin regulatory light chain (FhrMRLC) in Adjuplex® adjuvant against the infection with F. hepatica in rats. Four groups of 15 animals each were used for the study, one group was immunized with the recombinant F. hepatica MRLC in Adjuplex® adjuvant and the other groups remained as adjuvant, positive and negative control groups. The parasitological study showed that a statistically significant reduction of 65.1% and 82.1% in fluke burden and fecal egg count, respectively, was detected in vaccinated animals. In addition, vaccination with FhrMRLC induced a well-defined humoral and cellular immune response characterized by a significant production of specific IgG and IL-2, IL-12, TNF-α and IFN-γ; which confirms the immunogenic capacity of the FhrMRLC.

Evolution of the magnetic properties in the antiferromagnet Ce2RhIn8 simultaneously doped with Cd and Ir.

We report the evolution of the magnetic properties of Ce2Rh1-xIrxIn8-yCdy single crystals. In particular, for Ce2Rh0.5Ir0.5In8 (TN=2.0K) and Ce2Rh0.5Ir0.5In7.79Cd0.21 (TN=4.2K), we have solved the magnetic structure of these compounds using single-crystal neutron magnetic diffraction experiments. Taking the magnetic structure of the Ce2RhIn8 heavy-fermion antiferromagnet as a reference, we have identified no changes in the q=12,12,0 magnetic wave vector; however, the direction of the ordered Ce3+ moments rotates toward the ab plane, under the influence of both dopants. By constraining the analysis of the crystalline electric field (CEF) with the experimental ordered moment's direction and high-temperature magnetic-susceptibility data, we have used a mean-field model with tetragonal CEF and exchange interactions to gain insight into the CEF scheme and anisotropy of the CEF ground-state wave function when Cd and Ir are introduced into Ce2RhIn8. Consistent with previous work, we find that Cd doping in Ce2RhIn8 tends to rotate the magnetic moment toward the ab plane and lower the energy of the CEF excited states' levels. Interestingly, the presence of Ir also rotates the magnetic moment towards the ab plane although its connection to the CEF overall splitting evolution for the y = 0 samples may not be straightforward. These findings may shed light on the origin of the disordered spin-glass phase on the Ir-rich side of the phase diagram and also indicate that the Ce2MIn8 compounds may not follow exactly the same Rh-Ir CEF effects trend established for the Ce2MIn5 compounds.

Tuning the crystalline electric field and magnetic anisotropy along the CeCuBi2-xSbx series.

We have performed X-ray powder diffraction, magnetization, electrical resistivity, heat capacity and inelastic neutron scattering (INS) to investigate the physical properties of the intermetallic series of compounds CeCuBi2-xSbx. These compounds crystallize in a tetragonal structure with space group P4∕nmm and present antiferromagnetic transition temperatures ranging from 3.6 K to 16 K. Remarkably, the magnetization easy axis changed along the series, which is closely related to the variations of the tetragonal crystalline electric field (CEF) parameters. This evolution was analyzed using a mean field model, which included an anisotropic nearest-neighbor interactions and the tetragonal CEF Hamiltonian. We obtained the CEF parameters by fitting the magnetic susceptibility data with the constraints given by the INS measurements. More broadly, we discuss how this CEF evolution can affect the Kondo physics and the search for a superconducting state in this family.

Electronic and magnetic properties of stoichiometric CeAuBi2.

We report the electronic and magnetic properties of stoichiometric CeAuBi2 single crystals. At ambient pressure, CeAuBi2 orders antiferromagnetically below a Néel temperature (TN ) of 19 K. Neutron diffraction experiments revealed an antiferromagnetic propagation vector τ ^ = [ 0 , 0 , 1 ∕ 2 ] , which doubles the paramagnetic unit cell along the c axis. At low temperatures several metamagnetic transitions are induced by the application of fields parallel to the c axis, suggesting that the magnetic structure of CeAuBi2 changes as a function of field. At low temperatures, a linear positive magnetoresistance may indicate the presence of band crossings near the Fermi level. Finally, the application of external pressure favors the antiferromagnetic state, indicating that the 4f electrons become more localized.

Bilateral cleft lip and palate without premaxillary protrusion is associated with lethal aneuploidies.

OBJECTIVE: To investigate the clinical implications of two categories of fetal bilateral cleft lip and palate (BCLP): with premaxillary protrusion and with a flattened profile. METHODS: This was a retrospective study of fetuses with a prenatal diagnosis of BCLP at the Department of Obstetrics and Gynecology of the University of Bologna in the period 1991-2005. RESULTS: BCLP was diagnosed prenatally in 14 cases (mean gestational age at diagnosis, 21 (range, 12-36) weeks). In nine of these, there was a premaxillary pseudomass; in the remaining five, the profile was flat. Associated structural and/or chromosomal anomalies were found in two of the nine with a premaxillary pseudomass and in all five of those with a flat profile (P = 0.02). All fetuses with a flat profile had aneuploidies (three trisomy 18, one trisomy 13, one trisomy 8 mosaic), as did one of the nine with a premaxillary pseudomass. Eight of the pregnancies were terminated, including three of those with a premaxillary pseudomass and all five of those without. All continuing pregnancies resulted in live births, although one neonate affected by Krabbe's disease died shortly after birth. CONCLUSIONS: Our findings suggest that a third of cases of BCLP diagnosed in utero have a flat profile and these are at high risk of lethal aneuploidies.

Cervical length and risk of antepartum bleeding in women with complete placenta previa.

OBJECTIVE: To evaluate if cervical length predicts prepartum bleeding and emergency Cesarean section in cases of placenta previa. METHODS: Between September 2005 and September 2007, cervical length was measured by transvaginal ultrasound in women with complete placenta previa persisting into the third trimester of pregnancy. A complete follow-up of pregnancy was obtained in all cases. RESULTS: Overall, 59 women were included in the study group. The mean +/- SD gestational age at ultrasound was 30.7 +/- 2.7 weeks and the cervical length was 36.9 +/- 8.8 mm. Cesarean delivery was performed in all cases, at a mean gestational age of 34.7 +/- 2.3 weeks. Twenty-nine (49.1%) of the women presented prepartum bleeding and 12 (20.3%) required an emergency Cesarean section prior to 34 completed weeks due to massive hemorrhage. Cervical length did not differ significantly between cases with and those without prepartum bleeding (35.3 +/- 9.3 mm vs. 38.4 +/- 8.2 mm; P = 0.18), but was significantly shorter among patients who underwent emergency Cesarean section < 34 weeks due to massive hemorrhage compared with patients who underwent elective Cesarean section (29.4 +/- 5.7 mm vs. 38.8 +/- 8.5 mm; P = 0.0006). CONCLUSIONS: Transvaginal sonographic cervical length predicts the risk of emergency Cesarean section < 34 weeks in women with complete placenta previa.

Onset of motion of a partly hidden cylinder in a laminar shear flow.

In this paper, the onset of motion of an isolated cylinder partially exposed to a shear flow is experimentally investigated. The experiments are performed in a small narrow channel which provides a vertical shear layer flow whose sizes correspond with the channel width. The bottom of the channel is smooth except in the test zone, at long distance from the inlet, where the cylinder is placed with its principal axis perpendicularly directed to the main flow. The geometry of the channel bottom at the test zone is such that the cylinder is partially buried and presents different expositions to the incident flow. In this way, the geometrical constraints imposed by the sediment bed on a single particle in a natural sediment transport situation are reproduced in an idealized context. The results are interpreted in terms of the relation between the particle mobility parameter at the critical condition and the here defined particle burial degree with respect to the bed geometrical constraints beta . We emphasize the role played by this burial degree that is dependent on the particle exposure to the incident flow E and the resistance to the motion by mechanical contacts with its surroundings given by the so-called static pivot angle varphi .

Evidence of precursor orthorhombic domains well above the electronic nematic transition temperature in Sr(Fe1-x Co x )2As2.

Raman scattering, synchrotron x-ray diffraction, specific heat, resistivity and magnetic susceptibility measurements were performed in Sr(Fe1-x Co x )2As2 [[Formula: see text]] single crystals with superconducting critical temperature [Formula: see text] K and two additional transitions at 132 and 152 K observed in both specific heat and resistivity data. A quasielastic Raman signal with B 2g symmetry (tetragonal cell) associated with electronic nematic fluctuations is observed. Crucially, this signal shows maximum intensity at [Formula: see text] K, marking the nematic transition temperature. X-ray diffraction shows evidence of coexisting orthorhombic and tetragonal domains between [Formula: see text] and [Formula: see text] ∼ 152 K, implying that precursor orthorhombic domains emerge over an extended temperature range above [Formula: see text]. While the height of the quasielastic Raman peak is insensitive to [Formula: see text], the temperature-dependence of the average nematic fluctuation rate indicates a slowing down of the nematic fluctuations inside the precursor orthorhombic domains. These results are analogous to those previously reported for the LaFeAsO parent oxypnictide (Kaneko et al 2017 Phys. Rev. B 96 014506). We propose a scenario where the precursor orthorhombic phase may be generated within the electronically disordered regime ([Formula: see text]) as long as the nematic fluctuation rate is sufficiently small in comparison to the optical phonon frequency range. In this regime, the local atomic structure responds adiabatically to the electronic nematic fluctuations, creating a net of orthorhombic clusters that, albeit dynamical for [Formula: see text], may be sufficiently dense to sustain long-range phase coherence in a diffraction process up to [Formula: see text].

High-pressure studies on heavy-fermion antiferromagnet CeCuBi2.

We report in-plane electrical resistivity studies of CeCuBi2 and LaCuBi2 single crystals under applied pressure. At ambient pressure, CeCuBi2 is a c-axis Ising antiferromagnet with a transition temperature [Formula: see text] K. In a magnetic field applied along the c-axis at [Formula: see text] K a spin-flop transition takes place [Formula: see text] T. Applying pressure on CeCuBi2 suppresses T N at a slow rate. [Formula: see text] extrapolates to zero temperature at [Formula: see text] GPa. The critical field of the spin-flop transition [Formula: see text] displays a maximum of 6.8 T at [Formula: see text] GPa. At low temperatures, a zero-resistance superconducting state emerges upon the application of external pressure having a maximum T c of 7 K at 2.6 GPa in CeCuBi2. High-pressure electrical-resistivity experiments on the non-magnetic reference compound LaCuBi2 reveal also a zero resistance state with similar critical temperatures in the same pressure range as CeCuBi2. The great similarity between the superconducting properties of both materials and elemental Bi suggests a common origin of the superconductivity. We discuss that the appearance of this zero resistance state superconductivity may be related to the Bi layers present in the crystalline structure of both compounds and, therefore, could be intrinsic to CeCuBi2 and LaCuBi2, however further experiments under pressure are necessary to clarify this issue.

La dermatoscopia como herramienta para inferir el Breslow del melanoma / Dermoscopy as a Tool for Estimating Breslow Thickness in Melanoma

Antecedentes y objetivo: La incidencia del melanoma se ha incrementado significativamente y la forma más efectiva para disminuir su mortalidad es el diagnóstico precoz. La dermatoscopia aumenta la sensibilidad en el diagnóstico del melanoma, y por medio del análisis de las estructuras dermatoscópicas es posible estimar su grosor. Nuestro objetivo fue analizar la influencia del Breslow en las características dermatoscópicas del melanoma. Materiales y métodos: Estudio observacional de corte transversal. Se incluyeron pacientes con melanoma confirmado histológicamente y una imagen dermatoscópica del mismo. Se dividieron en tres grupos, melanoma in situ, melanoma fino (< 1 mm de Breslow) y melanoma grueso (≥ 1 mm de Breslow), y se analizaron el sexo, la edad, la localización, las características histológicas y las características dermatoscópicas. Resultados: Se analizaron 215 pacientes, 88 con melanoma in situ, 73 con melanoma fino y 54 con melanoma grueso. Las estructuras dermatoscópicas que incrementaron su frecuencia a medida que aumentó el Breslow del melanoma fueron el velo azul blanquecino (p < 0,001), las estructuras blanco brillantes (p < 0,001) y las áreas rojo lechosas (p < 0,003). Por otro lado, las líneas anguladas disminuyeron su frecuencia a medida que se incrementó el Breslow (p < 0,002). Conclusiones: La evaluación dermatoscópica tiene un importante rol, no solo en la precisión diagnóstica de las lesiones pigmentadas, sino también en ayudarnos a estimar el grosor preoperatorio del melanoma (AU)

Background and objective: The incidence of melanoma has increased significantly, and early diagnosis is the most effective way to reduce associated deaths. Dermoscopy increases diagnostic accuracy in melanoma and analysis of dermoscopic structures can help in the estimation of tumor thickness. The aim of this study was to analyze the influence of Breslow thickness on the dermoscopic characteristics of melanoma. Material and methods: Observational, cross-sectional study of patients with histologically confirmed melanoma and dermoscopic images of the tumor. The patients were divided into three groups: melanoma in situ, thin melanoma (≥ 1 mm Breslow thickness), and thick melanoma (≥ 1 mm Breslow thickness). Age, sex, tumor location, and histologic and dermoscopic characteristics were analyzed in all cases. Results: We studied 215 patients: 88 with melanoma in situ, 73 with thin melanoma, and 54 with thick melanoma. The frequency of the following dermoscopic features increased with increasing Breslow thickness: the blue-white veil (p < 0.001), white shiny structures (p < 0.001), and milky-red areas (p < 0.003). Angulated lines, by contrast, became less common with increasing thickness (p < 0.002). Conclusions: Dermoscopy not only improves diagnostic accuracy for pigmented lesions but also helps in the preoperative assessment of Breslow thickness in melanoma (AU)

Combined external pressure and Cu-substitution studies on BaFe2As2 single crystals.

We report a combined study of external pressure and Cu-substitution on BaFe2As2 single crystals grown by the in-flux technique. At ambient pressure, the Cu-substitution is known to suppress the spin density wave (SDW) phase in pure BaFe2As2(T(SDW) ≈ 140 K) and to induce a superconducting (SC) dome with a maximum transition temperature T(c)(max) ≃ 4.2 K. This T(c)(max) is much lower than the T(c) ∼ 15-28 K achieved in the case of Ru, Ni and Co substitutions. Such a lower T(c) is attributed to a Cu(2+) magnetic pair-breaking effect. The latter is strongly suppressed by applied pressure, as shown herein, Tc can be significantly enhanced by applying high pressures. In this work, we investigated the pressure effects on Cu(2+) magnetic pair-breaking in the BaFe(2-x)Cu(x)As2 series. Around the optimal concentration (x(opd) = 0.11), all samples showed a substantial increase of T(c) as a function of pressure. Yet for those samples with a slightly higher doping level (over-doped regime), T(c) presented a dome-like shape with maximum T(c) ≃ 8 K. Remarkably interesting, the under-doped samples, e.g. x = 0.02 display a maximum pressure induced T(c) ≃ 30 K which is comparable to the maximum T(c)'s found for the pure compound under external pressures. Furthermore, the magnetoresistance effect as a function of pressure in the normal state of the x = 0.02 sample also presented an evolution consistent with the screening of the Cu(2+) local moments. These findings demonstrate that the Cu(2+) magnetic pair-breaking effect is completely suppressed by applying pressure in the low concentration regime of Cu(2+) substituted BaFe2As2.

Alternative splicing and differential targeting of fibroblast growth factor receptor 1 in the pregnant rat uterus.

Recent studies suggest that steroid effects on uterine cell proliferation may be moderated by polypeptide growth factors. We now provide evidence that high affinity fibroblast growth factor (FGF) receptors are present temporally and spatially in the pregnant rat uterus (days 4-6) to support the idea that basic FGF action occurs via binding to its high affinity FGF receptor 1 (FGFR1). Reverse transcription-polymerase chain amplification indicates that both the full-length transcript and an alternatively spliced messenger RNA are present in the uterus. Western immunoblot analysis confirms that rat uterine membrane proteins contain two receptor isoforms, and these receptors bind basic FGF with high affinity and specificity. Immunolocalization of FGFR1 revealed receptor-positive cells in both the uterine stroma and epithelia on days 4-6 of pregnancy. However, the receptor was differentially localized in the disparate cell types. The nuclei of stromal cells were positive for FGFR1, whereas epithelial cell nuclei were negative. Together, these results suggest that FGF signal transduction in uterine stromal cells is mediated by activation of FGFR1.

Interleukin-6 differentially stimulates haptoglobin production by peritoneal and endometriotic cells in vitro: a model for endometrial-peritoneal interaction in endometriosis.

Based on our previous observation that peritoneal endometriotic (PE) lesions synthesize in vivo substantially more haptoglobin (Hp) than related eutopic tissues, we hypothesized that this increase in Hp production was due to endometrial-peritoneal interactions. As interleukin-6 (IL-6) stimulates Hp in other tissues and is produced by endometrial cells, we tested our hypothesis by evaluating the effects of IL-6 on Hp production by PE cells, normal peritoneal (P) cells, and eutopic endometrial cells from women with (UE-E) and without endometriosis (UE-C) using semiquantitative RT-PCR and enzyme-linked immunoabsorbent assay. Endogenous production of IL-6 was also assessed. Treatment with human recombinant IL-6 and dexamethasone significantly increased Hp production by P or PE cells in a dose- and time-dependent manner (P < 0.05). Hp messenger ribonucleic acid was not detected in UE-E and UE-C cells in the absence or presence of IL-6 and dexamethasone. PE and UE-E cells expressed significantly more IL-6 messenger ribonucleic acid than P and UE-C cells (P < 0.05). Moreover, UE-E cells secreted 6 times more IL-6 protein than UE-C cells (P < 0.05). These findings support our hypothesis and suggest a novel endometrial-peritoneal interaction whereby locally synthesized IL-6 and Hp may participate in the establishment and persistence of peritoneal endometriosis.

Benzodiazepines in the brain. Their origin and possible biological roles.

Great progress has been made in the last 5 yr in demonstrating the presence of benzodiazepines (BDZs) in mammalian tissues, in beginning studies on the origin of these natural compounds, and in elucidating their possible biological roles. Many unanswered questions remain regarding the sources and biosynthetic pathways responsible for the presence of BDZs in brain and their different physiological and/or biochemical actions. This essay will focus on recent findings supporting that: (1) BDZs are of natural origin; (2) mammalian brain contains BDZs in concentrations ranging between 5 x 10(-10)-10(-8) M; (3) dietary source of BDZs might be a plausible explanation for their occurrence in animal tissues, including man; (4) the formation of BDZ-like molecules in brain is a possibility, experimentally supported; (5) BDZ-like molecules including diazepam and N-desmethyldiazepam are elevated in hepatic encephalopathy; and (6) natural BDZs in the brain are involved in the modulation of memory processes. Future studies using the full range of biochemical, physiological, behavioral, and molecular biological techniques available to the neuroscientist will hopefully continue to yield exciting and new information concerning the biological roles that BDZs might play in the normal and pathological functioning of the brain.

Immunocytochemical localization of the beta 2 subunit of the gamma-aminobutyric acidA receptor in the rat brain.

An antiserum to the beta 2 subunit of the rat gamma-aminobutyric acid (GABAA) receptor was prepared by immunizing a rabbit with a fusion protein expressed in bacteria. The fusion protein had the large, intracellular loop expanding between the putative M3 and M4 transmembrane domains of the beta 2 subunit fused to staphylococcal protein A (SPA). The antiserum immunoprecipitated both the solubilized and the affinity-purified GABAA receptors. The anti-beta 2 antibodies were affinity purified on immobilized beta 2 intracellular loop peptide. The antibodies recognized a 55-57 kDa peptide in immunoblots of either crude membranes from rat cerebral cortex or affinity-purified GABAA receptors from bovine cerebral cortex. Immunocytochemistry with the affinity-purified antibody has revealed for the first time the localization of the beta 2 subunit in the rat brain. A comparative study of the regional and cellular immunoreactivities of the affinity-purified anti-beta 2 antibody and the monoclonal antibody 62-3G1 (which recognizes both beta 2 and beta 3 subunits) is presented. The procedure described for generating and preparing specific anti-beta 2 subunit antibodies that are valuable for immunocytochemistry could be extended to other GABAA receptor subunits.

Total PSA, free PSA/total PSA ratio, and molecular PSA detection in prostate cancer: which is clinically effective and when?

OBJECTIVES: To ascertain when the serum determination of the free prostate-specific antigen (PSA)/total PSA (fPSA/tPSA) ratio is clinically useful, and whether the identification of PSA or prostate-specific membrane antigen (PSM) mRNA in circulating cells has diagnostic advantages over the determination of their protein product. METHODS: fPSA, tPSA, and the fPSA/tPSA ratio were determined in the sera of 50 men with benign nonprostatic urologic diseases (EPD), 112 patients with prostate cancer (PCa), and 218 with benign prostatic hyperplasia (BPH). mRNA was extracted from the circulating mononuclear cells of 13 EPD samples, 25 PCa samples, and 38 BPH samples. PSA and PSM mRNA signals were identified in these samples by means of reverse transcriptase-polymerase chain reaction. RESULTS: Overall, at a fixed specificity of 95%, the sensitivity of tPSA was 19% and that of the fPSA/tPSA ratio was 40% in distinguishing PCa from BPH. The fPSA/tPSA ratio allowed the discrimination of PCa from BPH with satisfactory sensitivity and specificity when considering patients less than 60 years of age (100% and 95%, respectively). PSA and PSM mRNA were positive in 1 and 7 of 13 EPD samples, 6 and 13 of 25 PCa samples, and 6 and 17 of 38 BPH samples. The Gleason score did not correlate with tPSA, the fPSA/tPSA ratio, PSA mRNA, or PSM mRNA. CONCLUSIONS: The serum determination of the fPSA/tPSA ratio is an excellent index of PCa for subjects younger than 60 years of age; the clinical utility of PSA mRNA identification in circulating cells needs to be validated by large follow-up studies, and the analysis of PSM mRNA seems to be of no clinical interest.