Formal Verification Integration Approach for DSML

International audienceThe application of formal methods (especially, model check- ing and static analysis techniques) for the verification of safety critical embedded systems has produced very good results and raised the inter- est of system designers up to the application of these technologies in real size projects. However, these methods usually rely on specific verifica- tion oriented formal languages that most designers do not master. It is thus mandatory to embed the associated tools in automated verification toolchains that allow designers to rely on their usual domain-specific modeling languages (DSMLs) while enjoying the benefits of these power- ful methods. More precisely, we propose a language to formally express system requirements and interpret verification results so that system designers (DSML end-users) avoid the burden of learning some formal verification technologies. Formal verification is achieved through trans- lational semantics. This work is based on a metamodeling pattern for executable DSML that favors the definition of generative tools and thus eases the integration of tools for new DSML

Plasmodium falciparum metacaspase PfMCA-1 triggers a z-VAD-fmk inhibitable protease to promote cell death.

Activation of proteolytic cell death pathways may circumvent drug resistance in deadly protozoan parasites such as Plasmodium falciparum and Leishmania. To this end, it is important to define the cell death pathway(s) in parasites and thus characterize proteases such as metacaspases (MCA), which have been reported to induce cell death in plants and Leishmania parasites. We, therefore, investigated whether the cell death function of MCA is conserved in different protozoan parasite species such as Plasmodium falciparum and Leishmania major, focusing on the substrate specificity and functional role in cell survival as compared to Saccharomyces cerevisae. Our results show that, similarly to Leishmania, Plasmodium MCA exhibits a calcium-dependent, arginine-specific protease activity and its expression in yeast induced growth inhibition as well as an 82% increase in cell death under oxidative stress, a situation encountered by parasites during the host or when exposed to drugs such as artemisins. Furthermore, we show that MCA cell death pathways in both Plasmodium and Leishmania, involve a z-VAD-fmk inhibitable protease. Our data provide evidence that MCA from both Leishmania and Plasmodium falciparum is able to induce cell death in stress conditions, where it specifically activates a downstream enzyme as part of a cell death pathway. This enzymatic activity is also induced by the antimalarial drug chloroquine in erythrocytic stages of Plasmodium falciparum. Interestingly, we found that blocking parasite cell death influences their drug sensitivity, a result which could be used to create therapeutic strategies that by-pass drug resistance mechanisms by acting directly on the innate pathways of protozoan cell death

PENGETAHUAN DAN TINDAKAN PETUGAS TENTANG CUCI TANGAN PAKAI SABUN DI PUSKESMAS 23 ILIR KOTA PALEMBANG

Latar Belakang : Petugas Kesehatan di tuntut untuk memiliki pengetahuan untuk melakukan tindakan yang benar sesuai Standa Operasional Prosedur (SOP). Dalam hal ini petugas harus memiliki tingkat pengetahuan dan tindakan yang baik sehingga Standar Operasional Prosedur tercapai, salah satu Standard Operasional Prosedur tersebut adalah Cuci Tangan Pakai Sabun(CTPS). Cuci Tangan Pakai Sabun (CTPS) adalah salah satu tindakan sanitasi dengan membersihkan tangan dan jari-jemari menggunakan air dan sabun untuk menjadi bersih. Semakin tinggi tingkat pengetahuan Petugas Kesehatan maka semakin baik tingkat tindakannya. Metode : Desain Penelitian adalah deskriptif analitik dengan metode pendekatan penelitian kuantitatif dan rancangan pendekatan Random Sampling. Pelaksanaan KTI ini di Puskesmas 23 Ilir Palembang mulai pada tanggal 14 sampai tanggal 17 Januari 2021. Perencanaan dengan memberikan Lembar Kuisioner, Sampel Penelitian 30 Orang Petugas Puskesmas yang berisikan pertanyaan tentang Pengetahuan dan Tindakan Petugas Puskesmas 23 Ilir tentang Cuci Tangan Pakai Sabun (CTPS).  Hasil : Dari hasil penelitian mengenai pengetahuan dan tindakan petugas tentang cuci tangan pakai sabun di Puskesmas 23 Ilir adalah pengetahuan Petugas Kesehatan Puskesmas 23 Ilir memiliki tingkat pengetahuan baik sebanyak 73% dan tindakan petugas kesehatan Puskesmas 23 Ilir memiliki tindakan baik sebanyak 73%. Kesimpulan : Di Hasil tersebut Pengetahuan Petugas Kesehatan Puskesmas 23 Ilir tentang Cuci Tangan Pakai Sabun dikategorikan baik 80% sedangkan dari hasil penelitian nilai tindakan tentang Cuci Tangan Pakai Sabun Petugas Kesehatan Puskesmas 23 Ilir di kategorikan baik 73%

Tilt Table Practice Improved Ventilation in a Patient with Prolonged Artificial Ventilation Support in Intensive Care Unit

Patients who are on prolonged ventilator support in critical care unit present wide variety of complications, which range from reduction in oxygen uptake to various musculoskeletal impairments. Early mobilization and rehabilitation are encouraged to manage these complications effectively. Use of tilt table to motivate early mobilization in the intensive care unit for ventilator practices is not a usual practice. However, this new technique has attracted involvements of clinicians and therapists for its therapeutic benefits to the patient. Herein we describe a case of a seventy eight-year-old male patient who suffered Motor car accident, and was on ventilator support in intensive care unit for more than one month. He underwent treatment using a tilt table protocol with other routine treatment, which benefited him based on clinical as well as physiological variables. For practitioners in intensive care units, this report may offer perceptivity into the alternate practice of early mobilization using tilt table, and for investigators it may promote interest for further studies

Implication of different domains of the Leishmania major metacaspase in cell death and autophagy

International audienceMetacaspases (MCAs) are cysteine peptidases expressed in plants, fungi and protozoa, with a caspase-like histidine-cysteine catalytic dyad, but differing from caspases, for example, in their substrate specificity. The role of MCAs is subject to debate: roles in cell cycle control, in cell death or even in cell survival have been suggested. In this study, using a Leishmania major MCA-deficient strain, we showed that L. major MCA (LmjMCA) not only had a role similar to caspases in cell death but also in autophagy and this through different domains. Upon cell death induction by miltefosine or H2O2, LmjMCA is processed, releasing the catalytic domain, which activated substrates via its catalytic dyad His/Cys and a proline-rich C-terminal domain. The C-terminal domain interacted with proteins, notably proteins involved in stress regulation, such as the MAP kinase LmaMPK7 or programmed cell death like the calpain-like cysteine peptidase. We also showed a new role of LmjMCA in autophagy, acting on or upstream of ATG8, involving Lmjmca gene overexpression and interaction of the C-terminal domain of LmjMCA with itself and other proteins. These results allowed us to propose two models, showing the role of LmjMCA in the cell death and also in the autophagy pathway, implicating different protein domains

Crystal structure of a Trypanosoma brucei metacaspase

Metacaspases are distantly related caspase-family cysteine peptidases implicated in programmed cell death in plants and lower eukaryotes. They differ significantly from caspases because they are calcium-activated, arginine-specific peptidases that do not require processing or dimerization for activity. To elucidate the basis of these differences and to determine the impact they might have on the control of cell death pathways in lower eukaryotes, the previously undescribed crystal structure of a metacaspase, an inactive mutant of metacaspase 2 (MCA2) from Trypanosoma brucei, has been determined to a resolution of 1.4 Å. The structure comprises a core caspase fold, but with an unusual eight-stranded β-sheet that stabilizes the protein as a monomer. Essential aspartic acid residues, in the predicted S1 binding pocket, delineate the arginine-specific substrate specificity. In addition, MCA2 possesses an unusual N terminus, which encircles the protein and traverses the catalytic dyad, with Y31 acting as a gatekeeper residue. The calcium-binding site is defined by samarium coordinated by four aspartic acid residues, whereas calcium binding itself induces an allosteric conformational change that could stabilize the active site in a fashion analogous to subunit processing in caspases. Collectively, these data give insights into the mechanistic basis of substrate specificity and mode of activation of MCA2 and provide a detailed framework for understanding the role of metacaspases in cell death pathways of lower eukaryotes