A Kinase-deficient Splice Variant of the Human JAK3 Is Expressed in Hematopoietic and Epithelial Cancer Cells

Signal transduction of cytokine receptors is mediated by the JAK family of tyrosine kinases. Recently, the kinase partners for the interleukin (IL)-2 receptor have been identified as JAK1 and JAK3. In this study, we report the identification of splice variants that may modulate JAK3 signaling. Three splice variants were isolated from different mRNA sources: breast (B), spleen (S), and activated monocytes (M). Sequence analysis revealed that the splice variants contain identical NH2-terminal regions but diverge at the COOH termini. Analyses of expression of the JAK3 splice isoforms by reverse transcriptase-polymerase chain reaction on a panel of cell lines show splice preferences in different cell lines: the S-form is more commonly seen in hematopoietic lines, whereas the B- and M-forms are detected in cells both of hematopoietic and epithelial origins. Antibodies raised against peptides to the B-form splice variant confirmed that the 125-kDa JAK3B protein product is found abundantly in hematopoietic as well as epithelial cells, including primary breast cancers. The lack of subdomain XI in the tyrosine kinase core of the B-form JAK3 protein suggests that it is a defective kinase. This is supported by the lack of detected autokinase activity of the B-form JAK3. Intriguingly, both the S and B splice isoforms of JAK3 appear to co-immunoprecipitate with the IL-2 receptor from HUT-78 cell lysates. This and the presence of multiple COOH-terminal splice variants coexpressed in the same cells suggest that the JAK3 splice isoforms are functional in JAK3 signaling and may enrich the complexity of the intracellular responses functional in IL-2 or cytokine signaling

The DACAPO-PESO campaign: Dynamics, Aerosol, Cloud and Precipitation Observations in the Pristine Environment of the Southern Ocean: An overview

This article gives an overview of the DACAPO-PESO field experiment, which has taken place in Punta Arenas, Chile, from November 2018 to November 2021, and showcases first exciting research results that have already emerged from it.In diesem Artikel wird ein Überblick über das DACAPO-PESO Experiment gegeben, welches von November 2018 bis November 2021 in Punta Arenas, Chile, stattgefunden hat. Außerdem werden erste spannende Forschungsergebnisse vorgestellt, die bereits daraus gewonnen wurden

The Janus kinases (Jaks)

The Janus kinase (Jak) family is one of ten recognized families of non-receptor tyrosine kinases. Mammals have four members of this family, Jak1, Jak2, Jak3 and Tyrosine kinase 2 (Tyk2). Birds, fish and insects also have Jaks. Each protein has a kinase domain and a catalytically inactive pseudo-kinase domain, and they each bind cytokine receptors through amino-terminal FERM (Band-4.1, ezrin, radixin, moesin) domains. Upon binding of cytokines to their receptors, Jaks are activated and phosphorylate the receptors, creating docking sites for signaling molecules, especially members of the signal transducer and activator of transcription (Stat) family. Mutations of the Drosophila Jak (Hopscotch) have revealed developmental defects, and constitutive activation of Jaks in flies and humans is associated with leukemia-like syndromes. Through the generation of Jak-deficient cell lines and gene-targeted mice, the essential, nonredundant functions of Jaks in cytokine signaling have been established. Importantly, deficiency of Jak3 is the basis of human autosomal recessive severe combined immunodeficiency (SCID); accordingly, a selective Jak3 inhibitor has been developed, forming a new class of immunosuppressive drugs

Tumor-Shed PGE2 Impairs IL2Rγc-Signaling to Inhibit CD4+ T Cell Survival: Regulation by Theaflavins

BACKGROUND:Many tumors are associated with decreased cellular immunity and elevated levels of prostaglandin E2 (PGE2), a known inhibitor of CD4+ T cell activation and inducer of type-2 cytokine bias. However, the role of this immunomodulator in the survival of T helper cells remained unclear. Since CD4+ T cells play critical roles in cell-mediated immunity, detail knowledge of the effect tumor-derived PGE2 might have on CD4+ T cell survival and the underlying mechanism may, therefore, help to overcome the overall immune deviation in cancer. METHODOLOGY/PRINCIPAL FINDINGS:By culturing purified human peripheral CD4+ T cells or Jurkat cells with spent media of theaflavin- or celecoxib-pre-treated MCF-7 cells, we show that tumor-shed PGE2 severely impairs interleukin 2 receptor gammac (IL2Rgammac)-mediated survival signaling in CD4+ T cells. Indeed, tumor-shed PGE2 down-regulates IL2Rgammac expression, reduces phosphorylation as well as activation of Janus kinase 3 (Jak-3)/signal transducer and activator of transcription 5 (Stat-5) and decreases Bcl-2/Bax ratio thereby leading to activation of intrinsic apoptotic pathway. Constitutively active Stat-5A (Stat-5A1 6) over-expression efficiently elevates Bcl-2 levels in CD4+ T cells and protects them from tumor-induced death while dominant-negative Stat-5A over-expression fails to do so, indicating the importance of Stat-5A-signaling in CD4+ T cell survival. Further support towards the involvement of PGE2 comes from the results that (a) purified synthetic PGE2 induces CD4+ T cell apoptosis, and (b) when knocked out by small interfering RNA, cyclooxygenase-2 (Cox-2)-defective tumor cells fail to initiate death. Interestingly, the entire phenomena could be reverted back by theaflavins that restore cytokine-dependent IL2Rgammac/Jak-3/Stat-5A signaling in CD4+ T cells thereby protecting them from tumor-shed PGE2-induced apoptosis. CONCLUSIONS/SIGNIFICANCE:These data strongly suggest that tumor-shed PGE2 is an important factor leading to CD4+ T cell apoptosis during cancer and raise the possibility that theaflavins may have the potential as an effective immunorestorer in cancer-bearer

Electroanalysis may be used in the Vanillin Biotechnological Production

This study shows that electroanalysis may be used in vanillin biotechnological production. As a matter of fact, vanillin and some molecules implicated in the process like eugenol, ferulic acid, and vanillic acid may be oxidized on electrodes made of different materials (gold, platinum, glassy carbon). By a judicious choice of the electrochemical method and the experimental conditions the current intensity is directly proportional to the molecule concentrations in a range suitable for the biotechnological process. So, it is possible to imagine some analytical strategies to control some steps in the vanillin biotechnological production: by sampling in the batch reactor during the process, it is possible to determine out of line the concentration of vanillin, eugenol, ferulic acid, and vanillic acid with a gold rotating disk electrode, and low concentration of vanillin with addition of hydrazine at an amalgamated electrode. Two other possibilities consist in the introduction of electrodes directly in the batch during the process; the first one with a gold rotating disk electrode using linear sweep voltammetry and the second one requires three gold rotating disk electrodes held at different potentials for chronoamperometry. The last proposal is the use of ultramicroelectrodes in the case when stirring is not possible

Factors that enable and constrain the internationalisation and Africanisation of Master of Public Health programmes in South African highereducation institutions

Background. Higher education worldwide is currently shaped by globalisation and internationalisation, while African and South African (SA) highereducation institutions (HEIs) are required to Africanise their curricula to equip students to become effective and responsive global citizens, with globally and locally relevant knowledge and skills.Objectives. To establish the extent to which curricula for Master of Public Health programmes (MPHPs) within schools of public health or faculties of health sciences in selected HEIs in SA are currently internationalised and Africanised.Methods. The study followed a multiple-case-study design. Semi-structured and focus-group interviews with course co-ordinators, lecturers and students provided data. The study was conducted at three HEIs in SA that offer an MPHP. Tesch’s interactive process of qualitative data coding and analysis was used.Results. According to academics, there is no clear understanding or working definition of concepts and processes such as internationalisation and Africanisation as they apply to their professional contexts. The institutions do not subscribe to policies regarding internationalisation and Africanisation either. Academics are uncertain whether curricula meet the requirements of internationalisation and Africanisation, while students consider the curricula to be internationalised and Africanised to the best of their institutions’ and lecturers’ abilities.Conclusion. There is an urgent need for curriculum transformation in SA, to ensure that the internationalisation and Africanisation of curricula occur. Curriculum transformation and the formalisation of the processes of internationalisation and Africanisation through policy changes and capacity building need to be forefronted

Do South Africans really understand the National Health Insurance scheme?: a survey of adults in three provinces

A study was conducted among adult residents using health services in three provinces of South Africa to determine their understanding of the new National Health Insurance (NHI). The study was carried out among adult respondents in Limpopo, KwaZulu-Natal and the Eastern Cape provinces. A cross-sectional descriptive study design was used to select a sample of 784 from all the three provinces using a two-stage systematic sampling design. Data were collected using mobile phone assisted personal interviews (MPAPI). The respondents had a poor understanding of what the NHI would pay for. In this study, 48.1% knew that the NHI fund would pay for medical expenses if a person got sick, and 45.7% knew that with health insurance, basic health requirement is ensured and that if one becomes ill, medical treatment would be paid by the NHI fund. Again, 50.9% of South African respondents did not understand how the NHI fund will pay for health care received. Public awareness campaigns about the NHI were generally good while the education campaigns to increase understanding of the scheme were inadequate and did not penetrate many communities where information about the NHI was sought. We recommend a comprehensive community consultation plan to increase understanding of the NHI using various strategies such as house to house campaigns, town hall-type of meetings, workplace consultations and road shows at significant sites in communities such as clinics, schools, mines, farms and pension paypoints.

Participatory action research and community participation: developing a people's policy for health in South Africa

Poster presentation at the 13th International Congress of Behavioral Medicine in Groningen, Netherlands, 21 Augus