Use of Anti-Glycopeptidolipid-Core Antibodies Serology for Diagnosis and Monitoring of Mycobacterium avium Complex Pulmonary Disease in the United States.

Background: There is an unmet need for rapid, accurate, and noninvasive assays for diagnosis and monitoring of Mycobacterium avium complex pulmonary disease (MAC-PD). We evaluated the diagnostic accuracy of an anti-glycopeptidolipid (GPL)-core immunoglobulin A (IgA) antibody test in a US cohort of MAC patients, and we described serial serology changes during antimicrobial therapy. Methods: We identified serum samples from MAC patients starting treatment at enrollment and control subjects with or without bronchiectasis within OHSU's NTM Biobank. We conducted diagnostic test accuracy. Changes in mean levels of anti-GPL-core IgA antibodies between 0 and 3, 6, or 12 months after treatment start were assessed using the Student's paired t test. Pearson's correlation coefficient was calculated for IgA antibody levels and Student paired t test measures. Results: We included 25 MAC patients and 18 controls. At baseline, IgA antibody concentrations in MAC patients (3.40 ± 6.77 U/mL) were significantly higher than in controls without bronchiectasis (0.14 ± 0.03 U/mL, P = .02). Sensitivity and specificity for MAC-PD in this population was 48% and 89% (cutoff point 0.7 U/mL), respectively. Among MAC patients starting antimicrobial therapy, mean IgA levels decreased 0.3202 U/mL (P = .86) at month 3, 0.8678 U/mL (P = .47) at month 6, and 1.9816 U/mL (P = .41) at 1 year. Quality of Life-Bronchiectasis Respiratory Symptom Scale improvement correlated with decreasing IgA titers after 12 months of treatment in MAC patients (r = -0.50, P = .06). Conclusions: Anti-GPL-core IgA antibody levels are relatively specific for MAC-PD and decrease with treatment. Larger studies are warranted to evaluate the role of IgA serology in monitoring treatment response or for disease relapse/reinfection.

Correlación del dolor y la temperatura corporal en sujetos con lumbalgia: un estudio preliminar / Correlation of pain and body temperature in subjects with low back pain: a preliminary study

Introducción: El manejo del dolor es un tema importante de abordaje fisioterapéutico. La lumbalgia es una de las principales causas de dolor y discapacidad en el mundo, por lo que se abordó su estudio utilizando la termografía como herramienta de imagen para la evaluación objetiva y EVA para su valoración subjetiva. Objetivo: Describir la correlación entre sujetos con lumbalgia (PL) y sujetos sin lumbalgia (PS) utilizando como variables la sensación subjetiva de dolor y valores termográficos. Material y métodos: Se realizó un estudio correlacional donde se evaluaron 10 sujetos divididos en 2 grupos, un grupo de sujetos con lumbalgia y un grupo sin lumbalgia. Se obtuvo la temperatura en grados centígrados de las regiones de interés de los termogramas, la cual se correlacionó con la calificación del dolor establecida por los sujetos inicialmente. Resultados: La totalidad de sujetos sin lumbalgia no evidenció variaciones anormales de temperatura, mostrando una musculatura sana. Por el contrario, 2 sujetos con lumbalgia mostraron variaciones térmicas anormales, indicio de probable daño muscular. Los otros 3 sujetos no presentaron relación entre el dolor y la respuesta termográfica obtenida. Conclusiones: La relación dolor-termografía mostró una correlación de Pearson negativa para la región abdominal y positiva muy baja para la región lumbar, con un índice de significación estadística mayor de 0,05 (IC), por lo cual en este estudio no se encontró relación entre el índice de dolor y la temperatura por regiones musculares. La termografía infrarroja demostró ser una herramienta capaz de detectar, en tiempo real, alteraciones térmicas que presentan las zonas anatómicas potencialmente lesionadas.

Introduction: Pain management is an important issue in a physiotherapeutic approach. Low back pain is one of the main causes of pain and disability in the world, and therefore we studied it using thermography as an imaging tool for objective evaluation and the visual analogue scale for subjective evaluation. Objective: To describe the correlation between subjects with low back pain (PL) and subjects without low back pain (PS) using the subjective sensation of pain and thermographic values as variables. Material and methods: A correlational study was carried out, evaluating 10 subjects divided into two groups, a group of subjects with low back pain and a group without low back pain. The temperature was recorded in degrees Celsius, using thermograms and marking thermal points to evaluate each area, which was correlated with the pain rating initially established by the subjects. Results: The subjects without low back pain did not show abnormal temperature variations, and showed healthy musculature. In contrast, 2 subjects with low back pain showed abnormal thermal variations, an indication of probable muscle damage. The other 3 subjects did not show a relationship between pain and thermographic response. Conclusions: The pain-thermography relationship showed a negative Pearson correlation for the abdominal region and an extremely low positive correlation for the lumbar region, with a statistical significance index greater than .05 (CI). Therefore, in this study no connection was found between pain index and temperature by muscle region. Infrared thermography proved to be a tool capable of detecting thermal alterations in real time, in potentially injured anatomical areas.

Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging.

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

3D printer waste, a new source of nanoplastic pollutants.

Plastics pollution has been recognized as a serious environmental problem. Nevertheless, new plastic uses, and applications are still increasing. Among these new applications, three-dimensional resin printers have increased their use and popularity around the world showing a vertiginous annual-sales growth. However, this technology is also the origin of residues generation from the alcohol cleaning procedure at the end of each printing. This alcohol/resin mixture can originate unintentionally very small plastic particles that usually are not correctly disposed, and as consequence, could be easily released to the environment. In this work, the nanoparticle generation from 3D printer's cleaning procedure and their physicochemical characterization is reported. Nano-sized plastic particles are easily formed when the resin residues are dissolved in alcohol and placed under UV radiation from sunlight. These nanoparticles can agglomerate in seawater showing an average hydrodynamic diameter around 1 µm, whereas the same nanoparticles remain dispersed in ultrapure water, showing a hydrodynamic diameter of ≈300 nm. The formed nanoparticles showed an isoelectric point close to pH 2, which can facilitate their interaction with other positively charged pollutants. Thus, these unexpected plastic nanoparticles can become an environmental issue and public health risk.

Whole genome comparative analysis of four Georgian grape cultivars.

Grapevine is the one of the most important fruit species in the world. Comparative genome sequencing of grape cultivars is very important for the interpretation of the grape genome and understanding its evolution. The genomes of four Georgian grape cultivars-Chkhaveri, Saperavi, Meskhetian green, and Rkatsiteli, belonging to different haplogroups, were resequenced. The shotgun genomic libraries of grape cultivars were sequenced on an Illumina HiSeq. Pinot Noir nuclear, mitochondrial, and chloroplast DNA were used as reference. Mitochondrial DNA of Chkhaveri closely matches that of the reference Pinot noir mitochondrial DNA, with the exception of 16 SNPs found in the Chkhaveri mitochondrial DNA. The number of SNPs in mitochondrial DNA from Saperavi, Meskhetian green, and Rkatsiteli was 764, 702, and 822, respectively. Nuclear DNA differs from the reference by 1,800,675 nt in Chkhaveri, 1,063,063 nt in Meskhetian green, 2,174,995 in Saperavi, and 5,011,513 in Rkatsiteli. Unlike mtDNA Pinot noir, chromosomal DNA is closer to the Meskhetian green than to other cultivars. Substantial differences in the number of SNPs in mitochondrial and nuclear DNA of Chkhaveri and Pinot noir cultivars are explained by backcrossing or introgression of their wild predecessors before or during the process of domestication. Annotation of chromosomal DNA of Georgian grape cultivars by MEGANTE, a web-based annotation system, shows 66,745 predicted genes (Chkhaveri-17,409; Saperavi-17,021; Meskhetian green-18,355; and Rkatsiteli-13,960). Among them, 106 predicted genes and 43 pseudogenes of terpene synthase genes were found in chromosomes 12, 18 random (18R), and 19. Four novel TPS genes not present in reference Pinot noir DNA were detected. Two of them-germacrene A synthase (Chromosome 18R) and (-) germacrene D synthase (Chromosome 19) can be identified as putatively full-length proteins. This work performs the first attempt of the comparative whole genome analysis of different haplogroups of Vitis vinifera cultivars. Based on complete nuclear and mitochondrial DNA sequence analysis, hypothetical phylogeny scheme of formation of grape cultivars is presented.

Liver X Receptor Agonist Modifies the DNA Methylation Profile of Synapse and Neurogenesis-Related Genes in the Triple Transgenic Mouse Model of Alzheimer's Disease.

The liver X receptor agonist, GW3965, improves cognition in Alzheimer's disease (AD) mouse models. Here, we determined if short-term GW3965 treatment induces changes in the DNA methylation state of the hippocampus, which are associated with cognitive improvement. Twenty-four-month-old triple-transgenic AD (3xTg-AD) mice were treated with GW3965 (50 mg/kg/day for 6 days). DNA methylation state was examined by modified bisulfite conversion and hybridization on Illumina Infinium Methylation BeadChip 450 k arrays. The Morris water maze was used for behavioral analysis. Our results show in addition to improvement in cognition methylation changes in 39 of 13,715 interrogated probes in treated 3xTg-AD mice compared with untreated 3xTg-AD mice. These changes in methylation probes include 29 gene loci. Importantly, changes in methylation status were mainly from synapse-related genes (SYP, SYN1, and DLG3) and neurogenesis-associated genes (HMGB3 and RBBP7). Thus, our results indicate that liver X receptors (LXR) agonist treatment induces rapid changes in DNA methylation, particularly in loci associated with genes involved in neurogenesis and synaptic function. Our results suggest a new potential mechanism to explain the beneficial effect of GW3965.

Comparing regional transcript profiles from maize primary roots under well-watered and low water potential conditions.

Regionally distinct elongation responses to water stress in the maize primary root tip have been observed in the past. A genetic basis for such differential responses has been demonstrated. Normalized bar-coded cDNA libraries were generated for four regions of the root tip, 0-3 mm (R1), 3-7 mm (R2), 7-12 mm (R3), and 12-20 mm (R4) from the root apex, and transcript profiles for these regions were sampled. This permitted a correlation between transcript nature and regional location for 15 726 expressed sequence tags (ESTs) that, in approximately equal numbers, derived from three conditions of the root: water stress (water potential: -1.6 MPa) for 5 h and for 48 h, respectively, and well watered (5 h and 48 h combined). These normalized cDNA libraries provided 6553 unigenes. An analysis of the regional representation of transcripts showed that populations were largely unaffected by water stress in R1, correlating with the maintenance of elongation rates under water stress known for R1. In contrast, transcript profiles in regions 2 and 3 diverged in well-watered and water-stressed roots. In R1, transcripts for translation and cell cycle control were prevalent. R2 was characterized by transcripts for cell wall biogenesis and cytoskeleton formation. R3 and R4 shared prevalent groups of transcripts responsible for defence mechanisms, ion transport, and biogenesis of secondary metabolites. Transcripts which were followed for 1, 6, and 48 h of water stress showed distinct region-specific changes in absolute expression and changes in regulated functions.

Identification of a missense mutation in the bovine ABCG2 gene with a major effect on the QTL on chromosome 6 affecting milk yield and composition in Holstein cattle.

We previously localized a quantitative trait locus (QTL) on chromosome 6 affecting milk fat and protein concentration to a 4-cM confidence interval, centered on the microsatellite BM143. We characterized the genes and sequence variation in this region and identified common haplotypes spanning five polymorphic sites in the genes IBSP, SPP1, PKD2, and ABCG2 for two sires heterozygous for this QTL. Expression of SPP1 and ABCG2 in the bovine mammary gland increased from parturition through lactation. SPP1 and all the coding exons of ABCG2 and PKD2 were sequenced for these two sires. The single nucleotide change capable of encoding a substitution of tyrosine-581 to serine (Y581S) in the ABCG2 transporter was the only polymorphism corresponding to the segregation status of all 3 heterozygous and 15 homozygous sires for the QTL in the Israeli and U.S. Holstein populations. The allele substitution fixed effects on the genetic evaluations of 335 Israeli sires were -341 kg milk, +0.16% fat, and +0.13% protein (F-value = 200). No other polymorphism gave significant effect for fat and protein concentration in models that also included Y581S. The allele substitution effects on the genetic evaluations of 670 cows, daughters of two heterozygous sires, were -226 kg milk, 0.09% fat, and 0.08% protein (F-value = 394), with partial dominance towards the 581S homozygotes. We therefore propose that Y581S in ABCG2 is the causative site for this QTL.

Role of FSH and epidermal growth factor (EGF) in the initiation of steroidogenesis in granulosa cells associated with follicular selection in chicken ovaries.

In chicken ovaries, one small yellow follicle (SYF) is selected daily from a pool of follicles of similar size and becomes a preovulatory follicle. FSH induces follicular growth and steroidogenesis. Epidermal growth factor (EGF), an intraovarian hormone, suppresses granulosa cell differentiation. This study demonstrates that recruitment of SYFs into the hierarchy of preovulatory follicles is associated with a change in steroidogenic activity in granulosa cells regulated, at least in part, by FSH and EGF. Abundance of P450 side-chain cleavage (P450scc) mRNA was higher in the smallest preovulatory follicle (F6) compared with SYF, whereas FSH and EGF receptor (FSHr and EGFr, respectively) mRNA abundance was similar. FSH increased P450scc mRNA abundance and progesterone secretion and decreased FSHr mRNA in cultured granulosa cells, whereas EGF attenuated or suppressed P450scc mRNA and decreased FSHr mRNA abundance. None of the hormones influenced EGFr mRNA abundance. When used in combination, EGF attenuated or suppressed the stimulatory effect of FSH on the expression of P450scc mRNA and production of progesterone in a dose-dependent manner. The results indicate that (1) selection is associated with an increase in P450scc mRNA; (2) FSH stimulates expression of P450scc mRNA and progesterone secretion in granulosa cells of SYF; and (3) induction of P450scc mRNA and progesterone secretion by FSH is attenuated or blocked by EGF.

Follicular development and steroid concentrations in cows with different levels of fertility raised under nutritional stress.

The aim of the present study was to characterize ovarian follicular development and steroid concentrations during postpartum and the estrous cycle of Brangus Ibagé cows (3/8 Nelore + 5/8 Aberdeen Angus) with different levels of fertility. Cows were classified as having high or low fertility according to the calving interval (CI). The average CI of the herd from which cows used in this study were selected was 404.6+/-5.44 and 711.2+/-20.89 days for the high and low fertility groups, respectively. Four cows of high fertility and five cows of low fertility had calves removed between 70 and 100 days after parturition. Ovarian activity was monitored daily by ultrasound for 16 days after calf removal. Days to emergency of the first follicular wave after calf removal, number of follicles with diameter >9 mm, growth rate of largest follicle, maximum diameter of largest follicle, length (days) and number of follicular waves were recorded. During this period, blood was collected daily for measurements of serum progesterone (P(4)) and estradiol (E(2)) concentrations. In another experiment, ovarian activity and P(4) and E(2) concentrations were examined during estrous cycle in five cows of high fertility and four cows of low fertility. Ovarian activity and steroid concentrations were assessed from the day prior to estrus to the 15th day of the estrous cycle (estrus = day 0). In postpartum cows of high fertility, the total number of follicles >5mm and the maximum diameter of the largest follicle were higher than in cows of low fertility (P < 0.05). Concentrations of P(4) and E(2) did not differ between groups in the postpartum cows. However, E(2) increased 5 days after calf removal (around 90 days of postpartum) in the high fertility group, followed by an increase in P(4) with average values indicating ovulation around 100 days postpartum. In cycling cows, the profile of follicular development was similar between cows of high and low fertility. There was no difference between groups for number of follicles >5mm, but the day effect was significant (P < 0.01). Plasma concentrations of P(4) and E(2) were similar in both groups. These data suggest that cows, from a population raised in the same environment have different fertility as a consequence of individual physiological characteristics.

[Cases of hydatidiform mole at the Nuevo Hospital Civil in Guadalajara]. / Mola hidatidiforme en el Nuevo Hospital Civil de Guadalajara.

Thirty nine patients with hydatiform mole with clinical, echosonographic and hormonal diagnosis, were analyzed, from March, 1991 to February 1993 at the Nuevo Hospital Civil de Guadalajara. Incidence was I out of 301 pregnancies; average age, 24.6 years; 35.8% were primigravidae; average amenorreha was 15.6 weeks and there were not complications in 84.6%. The most frequent symptoms were transvaginal bleeding, 92.3%; mole rests expulsion, 25.6% and hyperemesis, 23%. The uterine fundus was larger in 17 patients; smaller in 13; the same in 5 and there were no data for four cases. An uterine curettage was performed in 38 patients; there was hysterotomy performed also for important bleeding; and there were 11 patients transfused. The histopathologic results of hydatidiform mole were confirmed in 84.6%. Only 20% of the patients came back for a post-evacuation control treatment. These results suggest that the characteristics of mola pregnancy at the Nuevo Hospital Civil de Guadalajara are similar to what has been reported in the literature; with a delayed diagnosis and a poor follow up of cases.

Extracellular phosphorylation in Leishmania major and Leishmania mexicana during heat shock transformation.

The extracellular phosphorylation of exogenous substrates have been studied in two Leishmania isolates, L. mexicana and L. major, that differ in their capacity to transform from promastigotes to amastigote-like cells when submitted to heat shock condition. When submitted to heat shock both parasites showed an increase in their extracellular phosphorylation activity. L. major promastigotes that do not transform to amastigote-like cells at 37 degrees C, do not phosphorylate exogenous substrate when the culture is grown at 25 degrees C. In contrast, L. mexicana promastigotes, that transform to amastigote-like cells at 37 degrees C, showed a strong phosphorylation at this temperature. As response to heat shock exposure, the extracellular phosphorylation for L. major increases steadily reaching a maximum at 360 min. On the other hand, L. mexicana also show an increase in phosphorylation during transformation, but in this case the maximum was detected after 10 min. The results are discussed in relationship to the capacity of the parasite to survive once inside the vertebrate host.

Relationships between cell surface protease and acid phosphatase activities of Leishmania promastigote.

A correlation between the ratio of the cell surface protease activity to phosphatase activity and the complexity of the pattern of cell surface exposed polypeptides of Leishmania promastigotes was demonstrated for various strains grown under similar conditions. The ratio of the cell surface protease activity to acid phosphatase activity was high for L. major and L.b. panamensis and it correlates with the expression of a single polypeptide of 63 KDa on their cell surface. Intermediate and lower ratios of these enzymatic activities relate with more complex radio-iodinated patterns: two main bands in L.b. guyanensis (70 and 58 KDa) and L.b. braziliensis (72 and 60 KDa) and three main bands 65, 50, 27 KDa in all L.m. mexicana strains tested. Evidence is presented that the acid phosphatase located on the L.m. mexicana cell surface is not an artifact due to a secondary absorption of the secreted acid phosphatase from the culture medium. These results confirm the Leishmania antigen cell surface heterogeneity. The implications on the biology of Leishmania and the clinical manifestation of leishmaniasis are discussed.

Antigenic analysis of Leishmania isolates from Tachira state, Venezuela.

Studies of cutaneous leishmaniasis in 3 endemic foci in Tachira state, western Venezuela have revealed sympatric populations of parasites causing both cutaneous and mucocutaneous disease. Immunological techniques and measurement of protease/acid phosphatase activities have been used to detect species-specific parasite antigens from 3 isolates from Tachira. Identified antigens of particular interest had molecular masses of 100, 82, 66, 50 and 27 kDa, but there was a high degree of heterogeneity between the antigens of the Tachira isolates and other Venezuelan strains of Leishmania braziliensis and L. mexicana. This heterogeneity has implications concerning the selection of antigens for use in serodiagnosis of leishmaniasis.

The effect of tunicamycin on the protease activity of GP63 from Leishmania major.

The protease activity of gp63 from L. major was studied in relation to tunicamycin induced N-deglycosylation. It was found that after tunicamycin treatment, a N-deglycosylated product of gp63 with protease activity is present at the cell surface of Leishmania promastigote.

Biochemical evidence of the antigenic cell surface heterogeneity of Leishmania mexicana.

In the present study, an enzymatical and structural analysis of Leishmania mexicana cell-surface components was carried out, demonstrating that protease and acid phosphatase activities were present at the L. mexicana cell surface. These findings correlate with the expression of the main components detected on the surface of L. mexicana promastigotes: the 50-kDa component is responsible for the acid phosphatase activity, whereas glycoprotein 65 (gp65) was characterized as the structural polypeptide of the surface protease. Furthermore, the 50- and 65-kDa antigens were found to be structurally different, inasmuch as no homology was observed in their peptide digestion profiles. The results presented in this communication confirm heterogeneity in the expression of the surface components of L. mexicana promastigotes at both the structural and the biochemical level.

The heterogeneity of Leishmania cell-surface antigens.

A comparative study of the radioiodinated promastigote cell-surface antigens of Leishmania mexicana and L. major was carried out under reduced and nonreduced conditions by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography. Under reduced conditions, the cell surface of L. mexicana promastigotes showed three iodinated polypeptides with molecular weights of 65,000, 50,000 and 27,000 daltons, whereas L. major promastigotes displayed a single polypeptide of 63,000 daltons. Under nonreduced conditions, the radioiodinated cell-surface component of L. major shifted to a mol.wt. of 51,000 daltons, whereas only one of the three components of L. mexicana (mol.wt., 65,000 daltons) underwent a large shift (to 59,000 daltons). The different immunochemical nature of the L. mexicana cell-surface antigens was demonstrated by using different anti-Leishmania sera. The rabbit anti-promastigote serum immunoprecipitated mainly the 50,000- and 27,000-dalton L. mexicana cell-surface polypeptides, whereas the rabbit anti-amastigote serum as well as a serum from a patient with cutaneous leishmaniasis immunoprecipitated almost exclusively the 65,000-dalton polypeptide. Immunoblot studies using a rabbit antibody against the L. major deglycosylated major surface antigen gp63 confirmed the differences in nature of the 65,000- and 50,000-dalton cell-surface antigens of L. mexicana. The results obtained are discussed in the light of the differences in antigenic cell-surface expression among Leishmania isolates and their consequences in the development of a differential diagnosis of leishmaniasis.

Plasma membrane and cytoskeletal constituents in Leishmania mexicana.

The presence and the localization of actin, spectrin and ankyrin are studied by immunofluorescence and immunoblotting in Leishmania mexicana promastigotes growing in vitro. These proteins, amphitropic in nature, coexist both in soluble and insoluble forms. Our results demonstrate that the Triton insoluble form of these proteins constitutes beside tubulin the cytoskeletal scaffold of promastigotes in close association with the plasma membrane, the axoneme and the basal body of the parasite.