Abstract: Cellular bio-energetic metabolism and mitochondria are recognized as potential targets for anticancer agents, due to the numerous relevant peculiarities cancer cells exhibit. Jasmonates are anticancer agents that interact directly with mitochondria. Many types of cancer cells exhibit overexpression of the key glycolytic enzyme, hexokinase, and its excessive binding to mitochondria. These characteristics are considered to play a pivotal role in cancer cell growth rate and survival. The aim of this study was to identify mitochondrial molecular targets of jasmonates. Methods: Binding and detachment of hexokinase from mitochondria were determined by hexokinase immunochemical and activity determinations, surface plasmon resonance analysis and planar lipid bilayer voltage dependent anion channel (VDAC)-activity analysis. Hexokinase expression was modified using hexokinase-overexpressing transfectants and its mitochondrial association. Results:. We report that jasmonates bind to hexokinase and detach it from the mitochondria and its mitochondrial anchor? VDAC. Jasmonate-induced detachment from mitochondria occurs in various types of cancer cells including leukemia and solid tumors. Furthermore, the susceptibility of cancer cells and mitochondria to jasmonates is dependent on the expression of hexokinase, supporting a cause and effect relationship between jasmonate-induced hexokinase detachment and cell death. Conclusions: 1) Our findings provide an explanation for the selective effects of jasmonates on cancer cells. 2) This is the first demonstration of a cytotoxic mechanism based on direct interaction between an anticancer agent and hexokinase. 3) The proposed mechanism can serve to guide development of a novel class of small anticancer compounds that kill cancer cells selectively by inhibiting the hexokinase?VDAC interaction. All

Abstract: Previously was shown that Infrasound (IS) at 4 Hz (30dB) frequency had modulating effect on heart muscle contractility. The molecular mechanism underlying this influence is not clear yet. The purpose of the present work is to study the molecular mechanism of infrasound. We have studied the effect of 4Hz (30dB) infrasound on heart muscle contractility, and Na/K pomp activity. We also measured the intracellular concentration of Ca2+ ions by 45Ca, and the content of intracellular cyclic nucleotides (cAMP, cGMP). Methods: Removed snail (Helix pomatia) hearts were cannulated and suspended in bath with physiological solution (PS). PS was exposed to 4Hz (30dB) IS for 30 minutes. 45Ca uptake by muscles incubated in normal and IS-treated PS was measured by 45Ca isotope. 45Ca-uptake and intracellular cyclic nucleotides contents (cAMP and cGMP) were measured by Wallac 1450 liquid scintillation counter. Results: IS at 4 Hz (30dB) frequency increased the amplitude and frequency of heart muscle contractility as well as the intracellular Ca2+ ions concentration. By measuring the content of intracellular cyclic nucleotides (cAMP, cGMP) we observed that by the increase of intracellular Ca2+ concentration caused the increase of cGMP content and decrease of cAMP contents in the cells.

Abstract: Spinal muscular atrophy (SMA) is one of the most common autosomal recessive disorders, characterized by degeneration of anterior horn cells in the spinal cord, and leads to progressive muscular weakness and atrophy. SMA is clinically divided into four subtypes depending on age at onset and clinical course. Genetic linkage studies have mapped responsible genes for all clinical types of SMA to chromosome 5q13, homozygous deletion in SMN1 gene causes the disease but the clinical severity may be modified by copy number of homologous gene SMN2 as well as the extent of deletion at the SMN locus. In the present study, to elucidate the correlation between genotype and clinical severity in SMA patients, we analyzed the molecular genetics features of 92 Algerian patients with SMA, from 57 unrelated families. All patients fulfilled the diagnostic criteria of SMA as defined by the International SMA Consortium. Genomic DNA was extracted from peripheral blood following the conventional procedures. Deletions of exons 7 and 8 of the SMN gene were analyzed by an enzyme digestion assay. NAIP gene analysis was performed by PCR amplification of exon 4 and 5. SMN2 gene copy number analysis was carried out by the use of a quantitative PCR-based assay. Patients were classified into type I SMA (20 patients), type II (16 patients), Type III (53 patients) and type IV (3 patients). 43 of the 57 SMA families (75.43%) were homozygous for SMN1 deletion of exon7 and 8 (type I, type II, type III and type IV). NAIP exon 4 and 5 were deleted in 15 (4/14 type I, 2/10 type II, 9/31 type III, 0 type IV), of 57 SMA families. In all patients with a NAIP deletion of exon 4 and 5, there was also a SMN1 deletion of exon 7. Frequency of NAIP deletions were significantly higher in type I patients than in type II or III patients. Also SMN2 copy numbers were higher in SMA type IV and type III than in type I and type II. This increase in copy number of SMN2 gene may be responsible for the less severe form of SMA with late onset of symptoms. Our results are compatible with the data of the literature.

Abstract: Our Bedouin community of ~170,000 is characterized by ~60% consanguineous marriages and 8-9 children per couple, leading to a high incidence of autosomal recessive diseases. A comprehensive organization of community education, molecular research and clinical implementation has been set up. We have generated effective novel software that facilitates finding disease-associated loci using Affymetrix SNP arrays, and a unique bioinformatics tool (S2G = Syndrome to Gene) to identify the disease genes within the defined loci. Using these tools, we recently identified 14 novel disease genes, 5 of them for severe neurological diseases: infantile neuroaxonal dystrophy (INAD) is a storage disease caused by mutations in phopholipase PLA2G6; a severe recessive phenotype of mental retardation and cerebral palsy is due to a mutation in UQCRQ of mitochondrial complex 3; Birk Barel genomic imprinting mental retardation syndrome is due to mutation in potassium channel KCNK9; Lethal congenital arthrogryposis can be caused by mutations in either ERBB3 or PIP5K1C, which directs ERBB3 internalization. These findings, with dozens of novel mutations we found in known disease genes, are being implemented as free carrier testing, prenatal diagnoses and pre-implantation genetic diagnoses, leading to a dramatic two-fold reduction in infant mortality in our Bedouin community over the past 4 years.

Abstract: Stenosis and atresias of the external auditory canal (EAC) are rare conditions difficult to manage with success. The main reason to operate is severity of the hearing impairment and threatening cholesteatoma of the EAC. Some authors do not find classical surgical methods sufficient and they favor the use of KTP/532 laser, or larger surgical procedures. The proven ability of Mitomycin C (MMC) to inhibit fibroblasts in vitro has stimulated its use it in treatment to prevent stenosis and adhesion, also in otorhinolaryngology. The objective of the study was to evaluate the opening of the external auditory canal in fibrotic atresias (congenitally and secondary) and the hearing improvement, after the surgery and concomitant use of topical MMC. Methods: Ten patients, all together fourteen ears, with fibrotic external auditory canal (EAC) atresias due to chronic external otitis, post-traumatic, post irradiated, or congenital cause were included. During the surgical procedure ? meatoplasty only endaural approach was used, and 1 mL of MMC (0.4 mg/mL) was applied for 4 minutes to the EAC. In 5 ears the application of MMC was repeated one to six month later, than 1mg/mL concentration of MMC was used. During the application of MMC the tympanic membrane was protected with a thin layer of dry Gelfoam. Audiometric evaluation included pre and postoperative air-conduction thresholds (AC) and bone-conduction thresholds (BC). Results: Between 9 and 56 months after the surgery and concomitant application of MMC the microscope visual control of opening of EAC was assessed. The hearing improvement was observed by using preoperative and postoperative pure tone threshold audiograms (PTA). In 10 ears (72 %) the ear canals reminded open with a postoperative air-bone gap of 10 dB or less. No sensorineural hearing loss was detected after surgery. Conclusions: 1) MMC used on a limited number of ears during surgery was effective in preventing scaring in fibrosis ending with atresias of EAC. 2) No complications or sensorineural hearing loss were encountered after surgery and MMC application. 3) The results of our study are comparable with the published reports and according to the follow up period our outcome results with a success rate of 72% could be considered as final.