Proteomic Changes in Chick Brain Proteome Post Treatment with Lathyrus Sativus Neurotoxin, ß-N-Oxalyl-L-α,ß-Diaminopropionic Acid (L-ODAP): A Better Insight to Transient Neurolathyrism.

Neurolathyrism is a neurodegenerative disorder characterized by spastic paraplegia resulting from the excessive consumption of Lathyrus sativus (Grass pea). ß-N-Oxalyl-L-α,ß-diaminopropionic acid (L-ODAP) is the primary neurotoxic component in this pea. The present study attempted to evaluate the proteome-wide alterations in chick brain 2 hr and 4 hr post L-ODAP treatment. Proteomic analysis of chick brain homogenates revealed several proteins involved in cytoskeletal structure, signaling, cellular metabolism, free radical scavenging, oxidative stress and neurodegenerative disorders were initially up-regulated at 2 hr and later recovered to normal levels by 4 hr. Since L-ODAP mediated neurotoxicity is mainly by excitotoxicity and oxidative stress related dysfunctions, this study further evaluated the role of L-ODAP in apoptosis in vitro using human neuroblastoma cell line, IMR-32. The in vitro studies carried out at 200 µM L-ODAP for 4 hr indicate minimal intracellular ROS generation and alteration of mitochondrial membrane potential though not leading to apoptotic cell death. L-ODAP at low concentrations can be explored as a stimulator of various reactive oxygen species (ROS) mediated cell signaling pathways not detrimental to cells. Insights from our study may provide a platform to explore the beneficial side of L-ODAP at lower concentrations. This study is of significance especially in view of the Government of India lifting the ban on cultivation of low toxin Lathyrus varieties and consumption of this lentil.

ß-N-oxalyl-L-α, ß- diaminopropionic acid induces HRE expression by inhibiting HIF-prolyl hydroxylase-2 in normoxic conditions.

Hypoxia inducible factor (HIF)-1α, a subunit of HIF transcription factor, regulates cellular response to hypoxia. In normoxic conditions, it is hydroxylated by prolyl hydroxylase (PHD)-2 and targeted for proteosomal degradation. Drugs which inhibit PHD-2 have implications in conditions arising from insufficient blood supply. ß-ODAP (ß-N- oxalyl-L-α, ß- diaminopropionic acid), a non-protein excitatory amino acid present in Lathyrus sativus, is an α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor agonist known to activate conventional protein kinase C and stabilize HIF-1α under normoxic conditions. However, the mechanism of HIF-1α stabilization by this compound is unknown. In silico approach was used to understand the mechanism of stabilization of HIF-1α which revealed ß-ODAP interacts with key amino acid residues and Fe2+ at the catalytic site of PHD-2. These results were further corroborated with luciferase HRE (hypoxia response element) reporter system in HeLa cells. Different chemical modulators of PHD-2 activity and HIF-1α levels were included in the study for comparison. Results obtained indicate that ß-ODAP inhibits PHD-2 and facilitates HIF dependent HRE expression and hence, might be helpful in conditions arising from hypoxia.

S137 phosphorylation of profilin 1 is an important signaling event in breast cancer progression.

BACKGROUND: Profilins are actin-modulating proteins regulating many intracellular functions based on their multiple and diverse ligand interactions. They have been implicated to play a role in many pathological conditions such as allergies, cardiovascular diseases, muscular atrophy, diabetes, dementia and cancer. Post-translational modifications of profilin 1 can alter its properties and subsequently its function in a cell. In the present study, we identify the importance of phosphorylation of profilin 1 at serine 137 (S137) residue in breast cancer progression. METHODS/PRINCIPAL FINDINGS: We found elevated profilin 1 (PFN) in human breast cancer tissues when compared to adjacent normal tissues. Overexpression of wild-type profilin 1 (PFN-WT) in breast cancer MCF7 cells made them more migratory, invasive and adherent independent in comparison to empty vector transfected cells. Mutation in serine phosphorylation site (S137) of profilin 1 (PFN-S137A) significantly abrogated these properties. Mutation affecting actin-binding ability (PFN-R74E) of profilin 1 enhanced its tumorigenic function whereas mutation affecting its poly-L-proline binding function (PFN-H133S) alleviated these mechanisms in breast cancer cells. PFN-WT was found to activate matrix metalloproteinases by zymography, MMP2 and MMP9 in presence of PDBu (phorbol 12, 13 dibutyrate, PI3K agonist) to enhance migration and invasion in MCF7 cells while PFN-S137A did not. Phosphorylation increased migration and invasion in other mutants of profilin 1. Nuclear profilin levels also increased in the presence of PDBu. CONCLUSIONS: Previous studies show that profilin could be executing a dual role in cancer by either suppressing or promoting tumorigenesis in a context dependent manner. In this study we demonstrate for the first time that phosphorylation of profilin 1 at serine 137 enhances oncogenic properties in breast cancer cells. Inhibitors targeting profilin 1 phosphorylation directly or indirectly through inhibition of kinases that phosphorylate profilin could be valuable therapeutic agents that can alter its activity and thereby control the progression of cancer.

Comparison between lengthening over nail and conventional Ilizarov lengthening: a prospective randomized clinical study.

The aim of this study is to compare lengthening over an intramedullary nail to the conventional Ilizarov method with regard to percentage length increase, external fixation index, consolidation index and incidence of complications. This is a prospective randomized controlled study. Thirty-one limbs in 28 patients were included in the study; 15 were lengthened over an intramedullary nail, and 16 limbs were lengthened conventionally. The mean duration of external fixation in the lengthening over nail group was 52.2 days compared to 180.4 days in the conventional group. There was higher incidence of complications in the conventional method group. In comparison with conventional Ilizarov lengthening, lengthening over an intramedullary nail offers a shorter period of external fixation and fewer complications overall, but there is a high incidence of deep intramedullary infection which is serious.

Pelvic support osteotomy by Ilizarov's concept: Is it a valuable option in managing neglected hip problems in adolescents and young adults?

For evaluating pelvic support osteotomy as a salvage procedure in managing neglected hip problems in adolescents and young adults, PSO was performed for 20 hips in 20 patients (5 men and 15 women). The mean age was 21.5 years. The etiology was neglected developmental dysplasia of the hip in 9 patients, post-septic hip sequelae in 9 patients, and paralytic dislocation due to poliomyelitis in 2 patients. All patients were treated by two osteotomies: a proximal femoral osteotomy to support the pelvis and correct the flexion and rotational deformities of the hip and a distal varization and lengthening osteotomy. Final clinical evaluation was done 6 months after frame removal. The mean external fixation time was 6.4. Lengthening and mechanical axis parallelism was achieved in all patients. At the final follow-up and according to a predesigned scoring system, there were 7(35%) excellent results, 6(30%) good results, 7(35%) fair results, and no poor results. Hip reconstruction by Ilizarov's concept can be technically demanding and involving lengthy period wearing the frame but found to be a valuable salvage procedure for numerous neglected hip problems particularly in young patients.

Fluorescence studies on the interaction of hydrophobic ligands with Momordica charantia (bitter gourd) seed lectin.

The interaction of Momordica charantia (bitter gourd) seed lectin (MCL) with several nucleic acid bases has been investigated by monitoring changes induced in the protein fluorescence by ligand binding. Values of the binding constant, K(a) were obtained as 1.1 x 10(4), 1.56 x 10(4) and 2.2 x 10(3) M(-1) for adenine, cytosine and uracil, respectively. In addition, binding of 8-anilinonaphthalene 1-sulfonate (ANS) with MCL was investigated by fluorescence spectroscopy. Interaction with MCL at low pH results in a large enhancement of the fluorescence intensity of ANS with a concomitant blue shift in the emission lambda(max), whereas at neutral and basic pH changes in both fluorescence intensity and emission maximum were very small, clearly suggesting that the MCL-ANS interaction is stronger at lower pH values. When excited at 295 nm in the presence of ANS, the protein fluorescence decreased with a concomitant increase in the emission intensity of ANS, suggesting resonance energy transfer from the tryptophan residues of MCL to ANS. Gel filtration profiles of MCL at pH values 2.0 and 7.4 are similar indicating that the tetrameric nature of MCL is retained even at low pH. Addition of lactose or adenine to MCL-ANS mixture did not alter the change in ANS fluorescence suggesting that lactose, adenine and ANS bind to MCL at independent and non-interacting sites. These results are relevant to understanding the functional role of MCL in the parent tissue.

Thermodynamic analysis of porphyrin binding to Momordica charantia (bitter gourd) lectin.

Owing to the use of porphyrins in photodynamic therapy for the treatment of malignant tumors, and the preferential interaction of lectins with tumor cells, studies on lectin-porphyrin interaction are of significant interest. In this study, the interaction of several free-base and metalloporphyrins with Momordica charantia (bitter gourd) lectin (MCL) was investigated by absorption spectroscopy. Difference absorption spectra revealed that significant changes occur in the Soret band region of the porphyrins on binding to MCL. These changes were monitored to obtain association constants (Ka) and stoichiometry of binding. The tetrameric MCL binds four porphyrin molecules, and the stoichiometry was unaffected by the presence of the specific sugar, lactose. In addition, the agglutination activity of MCL was unaffected by the presence of the porphyrins used in this study, clearly indicating that porphyrin and carbohydrate ligands bind at different sites. Both cationic and anionic porphyrins bind to the lectin with comparable affinity (Ka =10(3)-10(5) m(-1)). The thermodynamic parameters associated with the interaction of several porphyrins, obtained from the temperature dependence of the Ka values, were found to be in the range: DeltaH degrees = -98.1 to -54.4 kJ.mol(-1) and DeltaS degrees =-243.9 to -90.8 J.mol(-1).K(-1). These results indicate that porphyrin binding to MCL is governed by enthalpic forces and that the contribution from binding entropy is negative. Enthalpy-entropy compensation was observed in the interaction of different porphyrins with MCL, underscoring the role of water structure in the overall binding process. Analysis of CD spectra of MCL indicates that this protein contains about 13%alpha-helix, 36%beta-sheet, 21%beta-turn, and the rest unordered structures. Binding of porphyrins does not significantly alter the secondary and tertiary structures of MCL.

Epidemiological patterns of scurvy among Ethiopian refugees.

In the Horn of Africa, scurvy is a serious public health problem for refugees who are dependent on standard relief food (cereals, legumes, and oil). To assess the risk factors and to quantify the potential magnitude of scurvy among these displaced communities, we reviewed data collected from 1985 to 1987 by relief programmes in five refugee camps in Somalia and one in the Sudan. Outbreaks of clinical scurvy occurred among refugees in all the camps from 3 to 4 months after their arrival. The incidence of scurvy in two camps was, respectively, 14% over a period of 4 months and 19.8% over a period of 18 months. Prevalences of scurvy estimated from random population samples in the six study camps ranged from 13.6% to 44%. The risk of developing scurvy increased significantly with the length of time that refugees had been in the camps and was also significantly higher among those who were older and among females, particularly those of childbearing age. The prevalence of scurvy among refugees was similar, irrespective of whether or not they had attended supplementary feeding programmes. The control measures that were implemented had a moderate and slow impact on the disease. In both Somalia and the Sudan the relief food distributed to the refugees was almost completely deficient in vitamin C, while the environment where the camps were located precluded an adequate supply of fresh food. To avoid scurvy among refugee communities in this area of Africa it is therefore recommended that vitamin C supplements be added to the relief food at an early stage of a crisis.

Epidemiological patterns of scurvy among Ethiopian refugees

In the Horn of Africa, scurvy is a serious public health problem for refugees who are dependent on standard relief food (cereals, legumes, and oil). To assess the risk factors and to quantify the potential magnitude of scurvy among these displaced communities, we reviewed data collected from 1985 to 1987 by relief programmes in five refugees camps in Somalia and one in the Sudan. Outbreaks of clinical scurvy occurred among refugees in all the camps from 3 to 4 months after their arrival. The incidence of scurvy in two camps was, respectively, 14 per cent over a period of 4 months and 19.8 per cent over a period of 18 months. Prevalences of scurvy estimated from random population samples in the six study camps ranged from 13.6 per cent to 44 per cent. The risk of developing scurvy increased significantly with the length of time that refugees had been in the camps and was also significantly higher among those who were older and among females, particularly those of chidbearing age. The prevalence of scurvy among refugees was similar, irrespective of whether or not they had attended supplementary feeding programmes. The control measures that were implemented had a moderate and slow impact on the disease. In both Somalia and the Sudan the relief food distributed to the refugees was almost completely deficient in vitamin C ...(AU)