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Discusses how computer-mediated collaboration enables distant researchers to hold a conference and then collaborate on a single document. Roots of network-mediated collaboration; Current software such as `desktop conferencing' software developed by Group Technologies, and a Virtual Notebook System at Baylor College of Medicine; Electronic journal plans; Future of collaborative technology. INSET: Networks for thinking in cliques?, by A. Anderson..

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Abstract: Sarcocystis infections were found for the first time in the muscles of 3 of 3 gray wolves (Canis lupus) from Minnesota. Two kinds (thin-walled and thick-walled) of sarcocysts were detected, based on the appearance of the sarcocyst wall. In wolf 1, sarcocysts were thin-walled (<0.5 μm), and without any visible protrusions. Ultrastructurally, the sarcocyst wall was type 1a and identical to Sarcocystis svanai of the domestic dog (Canis familiaris). The second kind of sarcocyst, with a relatively thicker (>1 μm) sarcocyst wall, was detected in wolves 2 and 3. Ultrastructurally, the sarcocyst wall had undulating, pleomorphic villar protrusion of type 9c; these sarcocysts were identical to Sarcocystis caninum from the domestic dog. Molecularly, the 2 Sarcocystis species were characterized using 18S, 28S, COI, ITS-1, and rpoB genetic markers. All these markers showed 100% identity to either of the 2 species previously described from the domestic dog. The thick-walled sarococyst corresponded to Sarcocystis caninum, whereas the thin-walled sarcocyst corresponded to Sarcocystis svanai., (© American Society of Parasitologists 2024.)

The Minnesota, US moose population has declined dramatically since the 1990s. All 54 carcasses of moose that died of unknown cause or were euthanized by gun shot by tribal or Department of Natural Resources personnel because of perceived signs of illness between 2003 and 2013 and eight carcasses of moose that died from vehicular accidents between 2009 and 2013 were submitted to the Minnesota Veterinary Diagnostic Laboratory and included in our study. The majority of the animals were underweight or cachectic (n = 53; 85%). Neural migration presumably by Parelaphostrongylus tenuis was a common finding (n = 28; 45%). Moderate to marked Dermacentor albipictus ("winter tick") ectoparasitism with widespread alopecia was the cause or a contributing cause of death in 14 (23%) cases in which grossly apparent anemia was associated with exhaustion of hepatic iron stores. Hepatic lesions associated with Fascioloides magna were common (n = 37; 60%) but were unlikely to be the cause of death. Environmental factors favoring winter tick survival, habitat expansion of white-tailed deer (Odocoileus virginianus), and the survival of terrestrial and aquatic snails (serving as intermediate hosts for P. tenuis and F. magna), might contribute to the seemingly severe parasitic burden in Minnesota's moose population.

Ingestion of products containing Chinese star anise (Illicium verum) fruits contaminated or adulterated with Japanese star anise (Illicium anisatum) fruits can cause poisoning due to the neurotoxin anisatin that is present in Japanese star anise. Thus a rapid, simple and unambiguous distinction between the morphologically similar Chinese star anise and toxic Japanese star anise fruits is important for guaranteeing food safety. After adding ~200 μL of methanol to one star anise carpel placed at 7-10mm from the inlet of a mass spectrometer and applying a potential of ~5 kV to the carpel, an electrospray is created. The formation of the electrospray is immediate, robust and stable and lasts for at least a minute. The presence or absence of anisatin could be monitored by orbitrap high resolution mass spectrometry (HRMS) in negative mode by observing the [M-H](-) ion at m/z 327.1074 (C15H19O8) or in positive mode the [M+K](+) ion at m/z 367.079 (C15H20KO8). Several parameters like wetting solvent, voltage, distance and set-up were optimised. The anisatin signal was ~250 times higher in Japanese than in Chinese star anise. An existing Direct Analysis in Real Time (DART) HRMS for anisatin was used for benchmarking. Alternatively a linear ion trap mass spectrometer could be used in negative selective reaction monitoring (SRM) mode albeit with lower selectivity than the HRMS method. The transition of the [M-H](-) ion at m/z 327 to the fragment at m/z 265 was monitored. Direct plant spray and DART ionisation are both robust and provided the same yes/no answer in seconds without any prior sample preparation. Compared with the DART-HRMS procedure, the direct plant spray method is simpler in terms of equipment, yields a more stable signal, does not require heating of the sample but is slightly less selective and requires working with high voltages., (Copyright © 2013 Elsevier B.V. All rights reserved.)

B cells are important in mucosal microbial homeostasis through their well-known role in secretory IgA production and their emerging role in mucosal immunoregulation. Several specialized intraintestinal B cell compartments have been characterized, but the nature of conventional B cells in the lamina propria is poorly understood. In this study, we identify a B cell population predominantly composed of surface IgM(+) IgD(+) cells residing in villi of the small intestine and superficial lamina propria of the large intestine, but distinct from the intraepithelial compartment or organized intestinal lymphoid structures. Small intestinal (villous) B cells are diminished in genotypes that alter the strength of BCR signaling (Bruton tyrosine kinase(xid), Galphai2(-/-)), and in mice lacking cognate BCR specificity. They are not dependent on enteric microbial sensing, because they are abundant in mice that are germfree or genetically deficient in TLR signaling. However, villous B cells are reduced in the absence of invariant NK T cells (Jalpha18(-/-) or CD1d(-/-) mice). These findings define a distinct population of conventional B cells in small intestinal villi, and suggest an immunologic link between CD1-restricted invariant NK T cells and this B cell population.

A recent and surprising body of research has linked changes in immune function to biologic and therapeutic targeting of cannabinoid receptors, which prototypically respond to delta-9 tetrahydrocannabinol. The peripheral cannabinoid receptor CB2 is highly expressed in immune cell types (macrophages, dendritic cells, and B cells), and pharmacologically alters their cytokine production and responsiveness. Accordingly, cannabinoid agonists can powerfully alter susceptibility to certain microbial infections, atherosclerosis, and cancer immunotherapy. What is unknown is the physiologic role of natural levels of endocannabinoids and their receptors in normal immune homeostasis. Galphai2-/- mice are deficient in the formation of certain B and T cell subsets and are susceptible to immune dysregulation, notably developing inflammatory bowel disease. A key issue is the identity of the Gi-coupled receptors relevant to this Galphai2-signaling pathway. We find that mice deficient in CB2, the Gi-coupled peripheral endocannabinoid receptor, have profound deficiencies in splenic marginal zone, peritoneal B1a cells, splenic memory CD4+ T cells, and intestinal natural killer cells and natural killer T cells. These findings partially phenocopy and extend the lymphocyte developmental disorder associated with the Galphai2-/- genotype, and suggest that the endocannabinoid system is required for the formation of T and B cell subsets involved in immune homeostasis. This noncompensatable requirement for physiologic function of the endocannabinoid system is novel. Because levels of endocannabinoids are highly restricted microanatomically, local regulation of their production and receptor expression offers a new principle for regional immune homeostasis and disease susceptibility, and extends and refines the rationale for CB2-targeted immunotherapy in immune and inflammatory diseases.

Purpose: As kinase inhibitors transition from the laboratory to patients, it is imperative to develop biomarkers that can be used in the clinic. The primary objectives are to identify patients most likely to benefit from molecularly targeted therapies and to document modulation of the drug target. Constitutive activation of the phosphoinositide 3-kinase (PI3K) pathway and its downstream effectors, as a result of PTEN loss or by other mechanisms, occurs in a high proportion of prostate cancers, making it an ideal template for the design of clinical trials involving PI3K pathway inhibitors. Prostate cancers also present unique organ-specific challenges, in that tumors are heterogeneous and diagnostic tissue is extremely limited., Experimental Design: Working within these limitations, we have developed a set of immunohistochemical assays that define activation of the PI3K pathway in clinical samples., Results and Conclusions: Using both univariate and multivariate analyses, we show that loss of PTEN is highly correlated with the activation of AKT, and this, in turn, is associated with the phosphorylation of S6, one of its main effectors. These three antibodies are potentially able to define a molecular signature of PTEN loss and/or AKT pathway activation in prostate cancer.

Null mutation of the Galphai2 trimeric G protein results in a discrete and profound mucosal disorder, including inflammatory bowel disease (IBD), attenuation of IL-10 expression, and immune function polarized to Th1 activity. Genetic and adoptive transfer experiments have established a role for B cells and IL-10 in mucosal immunologic homeostasis and IBD resistance. In this study, we addressed the hypothesis that Galphai2 is required for the development of IL-10-producing B cells. Galphai2(-/-) mice were reduced in the relative abundance of marginal zone (MZ), transitional type 2 (T2), and B-1a B cells and significantly increased in follicular mature and B-1b B cells. Reconstitution of RAG2(-/-) mice with Galphai2(-/-) bone marrow induced an IBD-like colitis and a deficiency in absolute numbers of MZ, T2, and B-1 B cells. Thus, the Galphai2(-/-) genotype in colitis susceptibility and B cell development involved a cis effect within the hemopoietic compartment. In vitro, the B cell population of Galphai2(-/-) mice was functionally deficient in LPS-induced proliferation and IL-10 production, consistent with the exclusive capacity of T2 and MZ cell subpopulations for LPS responsiveness. In vivo, Galphai2(-/-) mice were selectively impaired for the IgM response to T-independent type II, consistent with the relative depletion of MZ and peritoneal B-1 subpopulations. Collectively, these results reveal a selective role for Galphai2 in MZ and B-1 B cell development. Disorders of this Galphai2-dependent process in B cell development may represent a mechanism for IBD susceptibility.

Follicular lymphoma (FL) is characterized by a continuous rate of relapse and transformation to a high-grade lymphoma, usually diffuse large B-cell lymphoma (DLBCL), associated with a dismal prognosis and a poor response to conventional chemotherapy. The progression of indolent to aggressive FL is accompanied by the successive accumulation of recurrent chromosomal defects, but the resultant alterations of gene expression are largely unknown. To expand the understanding of the pathogenesis of FL transformation, we initially performed oligonucleotide microarray analyses using Affymetrix HuFL chips on five cases with matched snap-frozen lymph nodes before and after transformation. Expression data were analyzed using the Affymetrix Microarray Suite 4.0 and Genespring 4.0. Thirty-six genes with increased expression and 66 genes with decreased expression associated with transformation were identified and functionally classified. The expression of differentially expressed genes was confirmed by real-time quantitative RT-PCR (QRT-PCR) using a total of seven matched pairs and an additional five FL and five unrelated DLBCL. In addition, selected genes were further analyzed by QRT-PCR or immunohistochemistry using a large, unrelated series of FL (grades 1 to 3) as well as transformed and de novo DLBCL (total of 51 samples). The microarray results correlated with the protein expression data obtained from samples at the time of initial diagnosis and transformation. Furthermore, the expression of 25 candidate genes was evaluated by QRT-PCR with a 78% confirmation rate. Some of the identified genes, such as nucleobindin, interferon regulatory factor 4, and tissue inhibitor of metalloproteinases 1, are already known to be associated with high-grade non-Hodgkin's lymphoma. Novel candidate genes with confirmed increased and decreased expression in transformed DLBCL include ABL2 and NEK2, and PDCD1 and VDUP1, respectively. In summary, this study shows that transformation of FL to DLBCL is associated with a distinct set of differentially expressed genes of potential functional importance.

Feeding experiments with specifically 13C-labeled glucose disclosed that the diterpenoid part of the striatals/striatins is formed via the mevalonate pathway, whereas the pentose moiety originates either via glucuronic acid (70%) or the pentose phosphate cycle (30%). Application of radioactively labeled herical to cultures of Hericium ramosum demonstrates the pivotal role of this cyathane-xyloside in striatal biosynthesis. Herical inhibits a large spectrum of fungi and bacteria and shows cytotoxic and hemolytic properties.

The authors assessed the impact of osteopathic manipulative treatment (OMT) as an adjunct to standard psychiatric treatment of women with depression. Premenopausal women with newly diagnosed depression were randomly assigned to either control (osteopathic structural examination only; n = 9) or treatment group (OMT; n = 8). Both groups received conventional therapy consisting of the antidepressant paroxetine (Paxil) hydrochloride plus weekly psychotherapy for 8 weeks. Attending psychiatrists and psychologists were blinded to group assignments. No significant differences existed between groups for age or severity of disease. After 8 weeks, 100% of the OMT treatment group and 33% of the control group tested normal by psychometric evaluation. No significant differences or trends were observed between groups in levels of cytokine production (IL-1, IL-10, IL-2, IL-4, and IL-6) or in levels of anti-HSV-1, anti-HSV-2, and anti-EBV antibody. There was no pattern to the osteopathic manipulative structural dysfunctions recorded. The findings of this pilot study indicate that OMT may be a useful adjunctive treatment for alleviating depression in women.

Prostate stem cell antigen (PSCA), a homologue of the Ly-6/Thy-1 family of cell surface antigens, is expressed by a majority of human prostate cancers and is a promising target for prostate cancer immunotherapy. In addition to its expression in normal and malignant prostate, we recently reported that PSCA is expressed at low levels in the transitional epithelium of normal bladder. In the present study, we compared the expression of PSCA in normal and malignant urothelial tissues to assess its potential as an immunotherapeutic target in transitional cell carcinoma (TCC). Immunohistochemical analysis of PSCA protein expression was performed on tissue sections from 32 normal bladder specimens, as well as 11 cases of low-grade transitional cell dysplasia, 21 cases of carcinoma in situ (CIS), 38 superficial transitional cell tumors (STCC, stages T(a)-T(1)), 65 muscle-invasive TCCs (ITCCs, stages T(2)-T(4)), and 7 bladder cancer metastases. The level of PSCA protein expression was scored semiquantitatively by assessing both the intensity and frequency (i.e., percentage of positive tumor cells) of staining. We also examined PSCA mRNA expression in a representative sample of normal and malignant human transitional cell tissues. In normal bladder, PSCA immunostaining was weak and confined almost exclusively to the superficial umbrella cell layer. Staining in CIS and STCC was more intense and uniform than that seen in normal bladder epithelium (P < 0.001), with staining detected in 21 (100%) of 21 cases of CIS and 37 (97%) of 38 superficial tumors. PSCA protein was also detected in 42 (65%) of 65 of muscle-invasive and 4 (57%) of 7 metastatic cancers, with the highest levels of PSCA expression (i.e., moderate-strong staining in >50% of tumor cells) seen in 32% of invasive and 43% of metastatic samples. Higher levels of PSCA expression correlated with increasing tumor grade for both STCCs and ITCCs (P < 0.001). Northern blot analysis confirmed the immunohistochemical data, showing a dramatic increase in PSCA mRNA expression in two of five muscle-invasive transitional cell tumors when compared with normal samples. Confocal microscopy demonstrated that PSCA expression in TCC is confined to the cell surface. These data demonstrate that PSCA is overexpressed in a majority of human TCCs, particularly CIS and superficial tumors, and may be a useful target for bladder cancer diagnosis and therapy.