Discovery of dual-target natural antimalarial agents against DHODH and PMT of Plasmodium falciparum: pharmacophore modelling, molecular docking, quantum mechanics, and molecular dynamics simulations.

Malaria is a lethal disease that claims thousands of lives worldwide annually. The objective of this study was to identify new natural compounds that can target two P. falciparum enzymes; P. falciparum Dihydroorotate dehydrogenase (PfDHODH) and P. falciparum phosphoethanolamine methyltransferase (PfPMT). To accomplish this, e-pharmacophore modelling and molecular docking were employed against PfDHODH. Following this, 1201 natural compounds with docking scores of ≤ -7 kcal/mol were docked into the active site of the second enzyme PMT. The top nine compounds were subjected to further investigation using MM-GBSA free binding energy calculations and ADME analysis. The results revealed favourable free binding energy values better than the references, as well as acceptable pharmacokinetic properties. Compounds ZINC000013377887, ZINC000015113777, and ZINC000085595753 were scrutinized to assess their interaction stability with the PfDHODH enzyme, and chemical stability reactivity using molecular dynamics (MD) simulation and density functional theory (DFT) calculations. These findings indicate that the three natural compounds are potential candidates for dual PfDHODH and PfPMT inhibitors for malaria treatment.

Expansion microscopy for the analysis of centrioles and cilia.

Centrioles are vital cellular structures that organise centrosomes and cilia. Due to their subresolutional size, centriole ultrastructural features have been traditionally analysed by electron microscopy. Here we present an adaptation of magnified analysis of the proteome expansion microscopy method, to be used for a robust analysis of centriole number, duplication status, length, structural abnormalities and ciliation by conventional optical microscopes. The method allows the analysis of centriole's structural features from large populations of adherent and nonadherent cells and multiciliated cultures. We validate the method using EM and superresolution microscopy and show that it can be used as an affordable and reliable alternative to electron microscopy in the analysis of centrioles and cilia in various cell cultures. LAY DESCRIPTION: Centrioles are microtubule-based structures organised as ninefold symmetrical cylinders which are, in human cells, ∼500 nm long and ∼230 nm wide. Centrioles assemble dozens of proteins around them forming centrosomes, which nucleate microtubules and organise spindle poles in mitosis. Centrioles, in addition, assemble cilia and flagella, two critically important organelles for signalling and motility. Due to centriole small size, electron microscopy has been a major imaging technique for the analysis of their ultrastructural features. However, being technically demanding, electron microscopy it is not easily available to the researchers and it is rarely used to collect large datasets. Expansion microscopy is an emerging approach in which biological specimens are embedded in a swellable polymer and isotopically expanded several fold. Physical separation of cellular structures allows the analysis of, otherwise unresolvable, structures by conventional optical microscopes. We present an adaptation of expansion microscopy approach, specifically developed for a robust analysis of centrioles and cilia. Our protocol can be used for the analysis of centriole number, duplication status, length, localisation of various centrosomal components and ciliation from large populations of cultured adherent and nonadherent cells and multiciliated cultures. We validate the method against electron microscopy and superresolution microscopy and demonstrate that it can be used as an accessible and reliable alternative to electron microscopy.

The KASH-containing isoform of Nesprin1 giant associates with ciliary rootlets of ependymal cells.

Biallelic nonsense mutations of SYNE1 underlie a variable array of cerebellar and non-cerebellar pathologies of unknown molecular etiology. SYNE1 encodes multiple isoforms of Nesprin1 that associate with the nuclear envelope, with large cerebellar synapses and with ciliary rootlets of photoreceptors. Using two novel mouse models, we determined the expression pattern of Nesprin1 isoforms in the cerebellum whose integrity and functions are invariably affected by SYNE1 mutations. We further show that a giant isoform of Nesprin1 associates with the ciliary rootlets of ependymal cells that line brain ventricles and establish that this giant ciliary isoform of Nesprin1 harbors a KASH domain. Whereas cerebellar phenotypes are not recapitulated in Nes1gSTOP/STOP mice, these mice display a significant increase of ventricular volume. Together, these data fuel novel hypotheses about the molecular pathogenesis of SYNE1 mutations and support that KASH proteins may localize beyond the nuclear envelope in vivo.

[Evaluation of quality of life (QOL) of Tunisians patients with COPD]. / Évaluation de la qualité de vie (QDV) des patients tunisiens atteints de bronchopneumopathie chronique obstructive (BPCO).

AIM: To test the Arabic version of the St. George's Hospital Respiratory Questionnaire (SGRQ) in stable Tunisian COPD patients. HYPOTHESES: A correlation coefficient between the post-bronchodilator FEV1 and the "Total" score of SGRQ higher than "-0.40", and QOL scores lower in COPD with "mild to moderate airway obstruction" than in those with "severe to very severe airway obstruction" will be in favor of a possible application of the Arabic version of the SGRQ in Tunisians COPD patients. METHODS: This is a prospective cross-sectional study including 50 clinically stable COPD patients. The Arabic version of the SGRQ was used. Four scores were calculated for the "Symptoms", "Activities", "Impacts" and "Total" components. Patients were divided into two groups according to the severity of their airway obstruction: "mild to moderate, n=30" and "severe to very severe, n=20". The correlation between the QOL "Total" score and postBD FEV1 was evaluated. RESULTS: The mean±SD of age, cigarette smoking and post-bronchodilator FEV1 were, respectively, 60±10, 61±36 pack-years and 55±20 %. The correlation between the post-bronchodilator FEV1 and QOL "Total" score was significant at "-0.65". "Symptoms", "Activities", "Impacts" and "Total" scores were significantly less altered in patients with "mild to moderate airway obstruction" than among those with "severe to very severe airway obstruction" (respectively, 52±24 vs. 77±17, 66±28 vs. 87±18, 48±25 vs. 70±23 and 54±22 vs. 76±17). CONCLUSION: The application of the Arabic version of the SGRQ in stable COPD patients gives reliable results.

Multifractal anisotropic swimming: the optimal foraging behaviour of grouper larvae.

It was hypothesized that the Malabar grouper Ephinephelus malabaricus larvae have developed search patterns adapted to the distribution of their prey to maximise their net energy intake per unit time. Analysis of the swimming behaviour of E. malabaricus larvae in both the presence and absence of Artemia sp. nauplii is presented to test this hypothesis. A method derived from turbulence studies (the moment function of the displacements) was used to characterize the behaviour. The results revealed that larval swimming pattern was multifractal (intermittent and long-range-correlated) and isotropic (i.e. uniform in all directions) in the presence of prey, but multifractal and anisotropic (i.e. more frequent long displacement on the vertical axis) in the absence of prey. It is suggested that the search behaviour observed in the absence of prey is an adaptive response to prey distribution pattern, which is often characterised by multifractality and anisotropy (i.e. larger patches on the horizontal axes). In the presence of prey, E. malabaricus shifted to intensive search behaviour. Other possible contributors to the observed patterns are discussed. It is concluded that multifractality and anisotropy of swimming patterns observed in the experiment are mainly explained in an optimal foraging theory framework.

Permanent cardiac pacing in a patient with persistent left superior vena cava and concomitant agenesis of the right-sided superior vena cava.

Persistent left superior vena cava (PLSVC) can be incidentally detected during pacemaker implantation from the left pectoral side. Optimal site pacing is technically difficult, and lead stability of the right ventricle (RV) can lead to such a situation. We describe a case of successful single-chamber pacemaker implantation in a 76-year-old woman with a PLSVC and concomitant agenesis of the right-sided superior vena cava, after failed attempts with the conventional procedure. The pacemaker had been working well after 12 months of follow-up.

[What are the predictive factors of the pain during the treatment of kidney stones by extra-corporeal shockwave lithotripsy?]. / Quels sont les facteurs prédictifs de la douleur lors du traitement des calculs rénaux par lithotritie extracorporelle ?

PURPOSE: Identify predictors for selecting patients who requires analgesia during lithotripsy. METHODS: This is a prospective study over a period of 13 months, 100 patients with kidney stones treated by an electromagnetic lithotripter (siemens; lithoskop) were selected. For the study of subjective pain caused by the ESWL at different times of the session, a visual analog scale (VAS) was used at different times (T) of the session (T0 before shots, T1 at 500 shots, T2 at 1500 shots, T end of treatment). A session was considered painless if VAS≤3. To identify predictors, were investigated association between pain and the different characteristics of patients, kidney stones and the shock wave specifications. RESULTS: The analytical study showed that pain was correlated with female gender, anxiety score, skin distance stones, parietal distance and the energy of the shock wave. While age, waist circumference, the circumstance found, the projection of stones on the rib and the number of shots had no impact on the level of pain. CONCLUSION: Our study showed that even with an electromagnetic lithotripter third generation; ESWL is still painful leading to the interruption of the session in 29% of cases. Four major predictors of pain leading to the use of sedo-analgesia early in the session were identified. LEVEL OF EVIDENCE: 3.

[Epidemiology of high risk pregnancy in Sousse, Tunisia]. / Profil épidémiologique des grossesses à risque à Sousse (Tunisie).

This descriptive study assessed the frequency of selected risk factors for high risk pregnancy (HRP) among women in Sousse region. All pregnant women (beyond 28 weeks gestation) giving birth in 4 public maternity hospitals between 15 February 2005 and 15 August 2005 and who had at least 1 risk factorwere enrolled. Data were collected within 24 hours of birth from obstetric and antenatal records and by interview. Of 4660 pregnant women, 1194 (25.6%) had at least 1 risk factor and were considered at-risk pregnancies. Mean age of the at-risk women was 31.3 (SD 5.4) years, 73% were urban residents, 38% had secondary education or higher and 75% were housewives. The mean number of risk factors was 1.5 per woman. The majority of women (59.3%) had 1 risk factor and 30.4% had 2. The mean number of prenatal visits was 4, 68.6% were seen by an obstetrician and 43% used the private sector. Certain factors were inadequately screened: age >35 years, parity >4, previous low birth weight and neonatal death, excessive fundal height and anaemia.

Epidemiology of high risk pregnancy in Sousse, Tunisia

This descriptive study assessed the frequency of selected risk factors for high risk pregnancy [HRP] among women in Sousse region. All pregnant women [beyond 28 weeks gestation] giving birth in 4 public maternity hospitals between 15 February 2005 and 15 August 2005 and who had at least 1 risk factor were enrolled. Data were collected within 24 hours of birth from obstetric and antenatal records and by interview. Of 4660 pregnant women, 1194 [25.6%] had at least 1 risk factor and were considered at-risk pregnancies. Mean age of the at-risk women was 31.3 [SD 5.4] years, 73% were urban residents, 38% had secondary education or higher and 75% were housewives. The mean number of risk factors was 1.5 per woman. The majority of women [59.3%] had 1 risk factor and 30.4% had 2. The mean number of prenatal visits was 4, 68.6% were seen by an obstetrician and 43% used the private sector. Certain factors were inadequately screened: age > 35 years, parity > 4, previous low birth weight and neonatal death, excessive fundal height and anaemia

Why subchondral bone in osteoarthritis? The importance of the cartilage bone interface in osteoarthritis.

Osteoarthritis is a whole joint disease characterised by the disappearance of the cartilage associated with subchondral bone sclerosis, formation of osteophytes and a mild inflammation of the synovial membrane. Although all these events have been independently studied, functional interactions between these different joint tissues should exist, especially between subchondral bone and cartilage. Moreover, recent studies show that cartilage and subchondral bone act as a single functional unit. This review highlights this novel concept.

Expression patterns of Notch receptors and their ligands in human osteoarthritic and healthy articular cartilage.

Notch pathway plays a pivotal role in cell fate determination. There is much interest surrounding its therapeutic potential, in osteoarthritis, but the expression profile of Notch-related molecules, as well as their relation with cartilage pathological parameters, remains unclear. The purpose of our study is to analyze the expression pattern of Notch family members, type II and type I collagen, in normal (healthy) and osteoarthritic human knee cartilage. Osteoarthritic cartilages were obtained from 3 patients undergoing a total knee replacement. Macroscopically normal cartilage was dissected from 3 human knees at the time of autopsy or surgery. Immunohistochemical staining was performed using Notch1,2,3 and 4, Delta, Jagged, type II collagen and type I collagen antibodies. In healthy cartilage, type II collagen was abundantly expressed while type I was absent. This latter increased proportionally to the osteoarthritic grade. Type II collagen expression remained intense in osteoarthritic cartilage. In healthy cartilage as well as in cartilage with minor lesions, Notch family member's proteins were not or just weakly expressed at the surface and in the cells. However, Notch molecules were over-expressed in osteoarthritic cartilage compared to healthy one. This expression pattern was different according to the cartilage zone and the severity of OA. Our data suggest that Notch signaling is activated in osteoarthritic cartilage, compared to healthy cartilage, with a much more abundant expression in the most damaged areas.

["Smoker's paradox" and reperfusion's strategy in acute myocardial infarction]. / "Paradoxe des fumeurs" et stratégie de reperfusion à la phase aiguë de l'infarctus du myocarde.

UNLABELLED: Previous studies have shown that smokers with acute myocardial infarction (AMI) treated by thrombolysis have lower mortality rates than nonsmokers, a phenomenon often termed "smoker's paradox". This "smoker's paradox" has been rarely studied in case of primary angioplasty. AIM OF THE STUDY: To evaluate the impact of smoking status on the early mortality of patients admitted with AMI with regard to the strategy of reperfusion (intravenous thrombolysis versus primary angioplasty). PATIENTS AND METHODS: Study undertaken from the Monsatir registry of ST elevation MI including 688 patients having had either a hospital or a prehospital thrombolysis (n=397) or a primary angioplasty (n=291). Among those patients, 482 (70.1%) were active smokers. RESULTS: In the thrombolysis group, the prevalence of hypertension, diabetes and anterior location of MI was significantly less among smokers. In the group primary angioplasty, only diabetes and hypertension were less frequent. The immediate mortality was significantly less among smokers in case of thrombolysis comparatively to non-smokers (5.3 vs 13%; p=0.008). By multivariate analysis, cardiogenic shock (p<0.0001), anterior MI (p=0.03) and active smoking (p=0.03) were independent predictive factors of mortality in case of thrombolysis. A trend toward a lower mortality among smokers was observed in the primary angioplasty group (10 vs 17.6%; p=0.07). CONCLUSION: "The smoker's paradox" seems to be observed mainly among patients having had thrombolysis.

Cellular and humoral responses induced by Leishmania histone H2B and its divergent and conserved parts in cutaneous and visceral leishmaniasis patients, respectively.

Leishmania histone H2B has been reported to be a promising candidate for both vaccination and serodiagnosis. We evaluated the cellular immune responses induced by H2B and its divergent amino-terminal (H2B-N) and conserved carboxy-terminal (H2B-C) regions in individuals with a history of Localized Cutaneous Leishmaniasis (LCL) due to Leishmania (L.) major. H2B induced significantly high PBMC proliferation and IFNgamma levels in LCL individuals whereas significantly lower proliferation and IFNgamma levels were observed with the divergent part of the protein. All proteins induced IL10 in LCL and healthy individuals. We also evaluated the humoral responses induced by these proteins in patients with Mediterranean Visceral Leishmaniasis (MVL) due to L. infantum. H2B and H2B-N were highly recognized by MVL sera. Our results show that the entire H2B protein is more efficient than its amino- and carboxy-terminal regions in inducing a dominant Th1 profile in cured LCL subjects and suggest that this protein may constitute a potential vaccine against leishmaniasis. Furthermore, H2B and H2B-N are interesting antigens for serodiagnosis of MVL.

Expression of Notch family members in cultured murine articular chondrocytes.

The Notch family is involved in cell differentiation during embryogenesis. Osteoarthritic chondrocytes undergo morphological and biochemical changes leading to the de-differentiation process. In the study reported here, we were interested in the involvement of the Notch pathway in murine articular chondrocyte de-differentiation. Articular chondrocytes were subjected to several cell culture passages and treated with or without a Notch inhibitor, N-[N-(3, 5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester (DAPT). Chondrocyte morphology was studied using optical microscopy. Immunocytochemistry and immunoblot were performed to study the expression of collagens and Notch family members. Without DAPT treatment, chondrocyte de-differentiation resulted in fibroblast-like morphology. This was confirmed by immunocytochemical staining and immunoblot analysis, which showed an increase in collagen type I (col I) and a decrease in collagen type II (col II) expression. With DAPT treatment, de-differentiation was delayed. Immunocytochemistry and immunoblot analysis showed during the first passages inhibition of col II expression, which then was re-instituted during the last passage, suggesting chondrocyte re-differentiation. In the study reported here, we showed that inhibition of the Notch receptor not only delayed the de-differentiation process, but also chondrocyte re-differentiation, which confirms the involvement of the Notch pathway in chondrocyte de-differentiation.

Chemical composition and antimicrobial activity of volatile compounds of Tamarix boveana (Tamaricaceae).

The chemical composition of the Tamarix boveana volatile oils obtained from the whole aerial part, flowers, leaves and stems by steam distillation was analysed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Sixty-two components were identified. Hexadecanoic acid (18.14%), docosane (13.34%), germacrene D (7.68%), fenchyl acetate (7.34%), Benzyl benzoate (4.11%) were found to be the major components in the whole aerial parts. This composition differed according to the tested part: 2.4 Nonadienal was the main compound in the flowers (12.13%) while germacrene D was the major component in leaves (31.43%) and hexadecanoic acid in the stems (13.94%). To evaluate in vitro antimicrobial activity, all volatile oils were tested against six Gram-positive and Gram-negative bacteria and four fungi. The T. boveana volatile oils exhibited an interesting antibacterial activity against all strains tested except Pseudomonas aeruginosa but no antifungal activity was detected.