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Bioinformatics and re-sequencing approaches were used for the discovery of sequence polymorphisms in Litopenaeus vannamei. A total of 1221 putative single nucleotide polymorphisms (SNPs) were identified in a pool of individuals from various commercial populations. A set of 211 SNPs were selected for further molecular validation and 88% showed variation in 637 samples representing three commercial breeding lines. An association analysis was performed between these markers and several traits of economic importance for shrimp producers including resistance to three major viral diseases. A small number of SNPs showed associations with test weekly gain, grow-out survival and resistance to Taura Syndrome Virus. Very low levels of linkage disequilibrium were revealed between most SNP pairs, with only 11% of SNPs showing an r2-value above 0.10 with at least one other SNP. Comparison of allele frequencies showed small changes over three generations of the breeding programme in one of the commercial breeding populations. This unique SNP resource has the potential to catalyse future studies of genetic dissection of complex traits, tracing relationships in breeding programmes, and monitoring genetic diversity in commercial and wild populations of L. vannamei. [ABSTRACT FROM AUTHOR]

Previous studies have uncovered several significant quantitative trait loci (QTL) relevant to meat colour traits mapped at the end of SSC17 in the pig. Furthermore, results released from the porcine genome sequencing project have identified genes underlying the entire QTL regions and can further contribute to mining the region for likely causative genes. Ten protein coding genes or novel transcripts located within the QTL regions were screened for single nucleotide polymorphisms (SNPs). Linkage mapping and association studies were carried out in the ISU Berkshire × Yorkshire (B × Y) pig resource family. The total length of the new SSC17 linkage map was 126.6 cM and additional markers including endothelin 3 ( EDN3) and phosphatase and actin regulator 3 ( PHACTR3) genes were assigned at positions 119.4 cM and 122.9 cM, respectively. A new QTL peak was noted at approximately 120 cM, close to the EDN3 gene, and for some colour traits QTL exceeded the 5% chromosome-wise significance threshold. The association analyses in the B × Y family showed that the EDN3 BslI and PHACTR3 PstI polymorphisms were strongly associated with the subjective colour score and objective colour reflectance measures in the loin, as well as average drip loss percentage and pH value. The RNPC1 DpnII and CTCFL HpyCH4III polymorphisms were associated with some meat colour traits. No significant association between CBLN4, TFAP2C, and four novel transcripts and meat colour traits were detected. The association analyses conducted in one commercial pig line found that both EDN3 BslI and PHACTR3 PstI polymorphisms were associated with meat colour reflectance traits such as centre loin hue angle and Minolta Lightness score. The present findings suggested that the EDN3 and PHACTR3 genes might have potential effects on meat colour in pigs, and molecular mechanisms of their functions are worth exploring. [ABSTRACT FROM AUTHOR]

Liver X receptor alpha (LXRA) and beta (LXRB) are members of the nuclear receptor subfamily and are important regulators of genes involved in lipid, fatty acid and glucose metabolism in liver, and adipose tissue as well as in skeletal muscle. To investigate whether the two LXR genes play a role in influencing lean and fat growth in pigs, we discovered and examined two polymorphisms in LXRA ( LXRA Bsl in exon 2, and LXRA HpyCH4 III in intron 8) and one polymorphism in LXRB ( LXRB Aci I in exon 5) for genetic linkage and association analyses. Linkage analyses using a three-generation resource family of a cross between the Berkshire and Yorkshire (BY) pig breeds assigned LXRA to SSC2 and LXRB to SSC6. Association analyses were carried out among those polymorphisms and traits evaluated in the BY F2 family and four pig commercial populations. These analyses indicated that the LXRA HpyCH4 III polymorphism was significantly associated with loin eye area and total lipid in individuals from the BY family. Significant associations were also found between Bsl I polymorphism in LXRA and boneless loin (%), as well as marbling score in one commercial line. The LXRB Aci I polymorphism was significantly associated with lean meat and fat content in the BY family and a number of the commercial lines examined. Our current findings suggested that LXRA and LXRB might have potential effects, especially for loin lean and fat content. [ABSTRACT FROM AUTHOR]

Automated acquisition of high resolution, light microscope images of cells is becoming a common requirement in modern proteomic and cellomic research. A prerequisite for such microscopy is fine focus tuning, commonly optimized by multiple exposures, followed by image sharpness analysis. We describe here an extremely fast and accurate laser autofocusing system with distinct advantages for large-scale cell-based screening. [ABSTRACT FROM AUTHOR]

The mode of interaction with macrophages of two ordered synthetic polypeptides (Tyr-Tyr-Glu-Glu)-poly(DLAla)—poly(Lys), (T-T-G-G)-A—L, and (Tyr-Glu-Tyr-Glu)-poly(DLAla)—poly(Lys), (T-G-T-G)-A—L, which differ in their requirements for T-B cell co-operation in the process of antibody production, was compared. The binding of the two radiolabelled antigens to the surface of peritoneal adherent cells, their uptake by the cells and the rate of their degradation were investigated. Macrophages were found to be capable of degrading both polypeptides with the same efficiency. (T-G-T-G)-A—L, the antigen which is less T-dependent, was bound to macrophage surfaces more readily than (T-T-G-G)-A—L, the T-dependent antigen, however, its uptake by the cells was found to be lower. Thus, (T-G-T-G)-A—L remains for a longer period in the form of a membrane bound polyvalent antigen. [ABSTRACT FROM AUTHOR]

The native secretion of rat seminal vesicles was found to contain about 290 mg of protein/ml. The ionic strength of the secretion was low (33 millimho. cm-2). Some protein (about 13 per cent) precipitated shortly after collection. The soluble proteins were resolved by electrophoresis into four very basic and two minor acidic fractions. Three of the proteins were eluted from a Sephadex G-200 column at a position indicating a molecular weight much in excess of 150,000 daltons; the others were eluted at a position intermediate between IgG and bovine serum albumin (BSA) markers. Ten bands were resolved in polyacrylamide gel electrophoresis following dissociation of the soluble proteins into subunits. Eight of these had molecular weights ranging between 18,500 and 100,000 Daltons. Antisera raised in rabbits against seminal vesicle fluid (SVF) formed, in immunoelectrophoresis (IE) six precipitin bands with SVF. They did not react at all with rat serum proteins, nor did antiserum to rat serum proteins react with SVF. A small amount of rat serum proteins was, however, detected by radioimmunoassay in the insoluble fraction. [ABSTRACT FROM AUTHOR]

SUMMARY Studies of the three-dimensional (3-D) organization of cell nuclei are becoming increasingly important for the understanding of basic cellular events such as growth and differentiation. Modern methods of molecular biology, including in situ hybridization and immunofluorescence, allow the visualization of specific nuclear structures and the study of spatial arrangements of chromosome domains in interphase nuclei. Specific methods for labelling nuclear structures are used to develop computerized techniques for the automated analysis of the 3-D organization of cell nuclei. For this purpose, a coordinate system suitable for the analysis of tri-axial ellipsoidal nuclei is determined. High-resolution 3-D images are obtained using confocal scanning laser microscopy. The results demonstrate that with these methods it is possible to recognize the distribution of visualized structures and to obtain useful information regarding the 3-D organization of the nuclear structure of different cell systems. [ABSTRACT FROM AUTHOR]

Vinculin is a 117 kDa microfilament-associated protein located at the cytoplasmic aspects of focal contacts and cell-cell adherens type junctions. In both sites, vinculin participates in the formation of a submembrane `plaque' structure which is responsible for the attachment of actin flaments to the plasma membrane. Vinculin consists of 1066 amino acids, which form a large 90 kDa globular head domain and a rod-like 29 kDa tail domain. The two domains are separated by several stretches of proline residues where the major proteolytic cleavage sites are located. The experimental procedure for isolation and purification of vinculin from smooth muscle has been developed and crystals of native vinculin suitable for X-ray analysis have been obtained. The homogeneity of the vinculin solution was analyzed prior to crystallization using dynamic light scattering. Crystals of vinculin have been obtained in buffer containing 2 mg ml-1 protein, 0.9 M ammonium sulfate, 0.1 M MES pH 6.5 using both the hanging-drop and sitting-drop vapour-diffusion methods. The crystals have the form of rhombic plates and grow to maximal dimensions of 0.3 × 0.3 × 0.05 mm in two weeks. Preliminary X-ray data show that the crystals diffract to 3.5 Å resolution at the X11 beamline of DESY and belong to the monoclinic space group P21. Crystal unit-cell parameters are estimated to be a = 57, b = 351, c = 70 Å, α = 90, β = 113, γ = 90°. [ABSTRACT FROM AUTHOR]

Chronic myeloid leukaemia (CML) management is complicated by treatment-emergent vascular adverse events seen with tyrosine kinase inhibitors (TKIs) such as nilotinib, dasatinib and ponatinib. Pleural effusion and pulmonary arterial hypertension (PAH) have been associated with dasatinib treatment. Endothelial dysfunction and impaired angiogenesis are hallmarks of PAH. In this study, we explored, at cellular and whole animal levels, the connection between dasatinib exposure and disruption of endothelial barrier integrity and function, leading to impaired angiogenesis. Understanding the mechanisms whereby dasatinib initiates PAH will provide opportunities for intervention and prevention of such adverse effects, and for future development of safer TKIs, thereby improving CML management., (© 2024 The Author(s). British Journal of Haematology published by British Society for Haematology and John Wiley & Sons Ltd.)

Introduction: People who inject drugs are 13 times more likely to die by suicide than the general population. Guidelines for responding to risk in this population are limited. Harm reduction services attended by people who inject drugs require targeted strategies to address the complexities of suicide risk among this population., Methods: Co-design, engaging health professionals and people with lived experience informed the study. Mixed methods were used to understand the experience of managing suicide risk among clients attending the Medically Supervised Injecting Centre (MSIC) in Sydney. A survey was administered to assess staff confidence in managing risk. Focus groups were conducted with health professionals and MSIC clients to explore experiences of suicide management, response and opportunities for improvement., Results: Half (N = 17) the MSIC staff surveyed reported over 10 years' experience working with this population. Confidence in managing suicide risk was low. Three key themes emerged from focus groups (N = 17): (i) Autonomy and the need to involve clients in the assessment process; (ii) Trust between clients and health professionals, and transparency in decision-making; and (iii) System barriers, described by health professionals as inadequate referral pathways for clients in distress, and by clients as negative experiences of care, including involuntary admission and not receiving medication (e.g. methadone)., Discussion and Conclusions: Revised assessment guidelines and a tailored safety plan were developed. These resources are also suitable for other alcohol and other drug services. The challenge in managing suicide risk in harm reduction services is balancing duty of care with staff-client relationships and client engagement., (© 2024 The Author(s). Drug and Alcohol Review published by John Wiley & Sons Australia, Ltd on behalf of Australasian Professional Society on Alcohol and other Drugs.)

A basic criterion for the diagnosis of multiple myeloma is plasma cell enumeration within the bone marrow (BM). This report showed that flow cytometry under-estimated the number of plasma cells in BM aspirates by an average of 60%, compared with morphological evaluation. The discrepancy was partially because BM smears contain cells associated with the lipid-enriched spicules. In contrast, flow cytometry is performed on the BM fluid, which is depleted of the lipid-adhesive plasma cells. This discrepancy may point to different plasma cell subpopulations associated with diverse niches within the BM.

The study of normal or malignant haematopoiesis requires the analysis of heterogeneous cell populations using multiple morphological and molecular criteria. Flow cytometry has the capacity to acquire multi-parameter information of large haematopoietic cell populations, utilizing various combinations of >200 molecular markers (clusters of differentiation, CD). However, current flow cytometry analyses are based on serial gating of two-parametric scatter plots--a process that is inherently incapable to discriminate all subgroups of cells in the data. Here we studied the cellular diversity of normal bone marrows (BM) using multi-dimensional cluster analysis of six-parametric flow cytometry data (four CD, forward scatter and side scatter), focusing mainly on the myeloid lineage. Twenty-three subclasses of cells were resolved, many of them inseparable even when examined in all possible two-parametric scatter plots. The multi-dimensional analysis could distinguish the haematopoietic progenitors according to International Society of Haematotherapy and Graft Engineering criteria from other types of immature cells. Based on the defined clusters, we designed a classifier that assigns BM cells in samples to subclasses based on robust six-dimensional position and extended shape. The analysis presented here can manage successfully both the increasing numbers of haematopoietic cellular markers and sample heterogeneity. This should enhance the ability to study normal haematopoiesis, and to identify and monitor haematopoietic disorders.