Synthesis,Biological Evaluation,and Molecular Docking of Novel Thiazoles and [1,3,4]Thiadiazoles Incorporating Sulfonamide Group as DHFR Inhibitors

2‐(1‐{4‐[(4‐Methylphenyl)sulfonamido]phenyl}ethylidene)thiosemicarbazide ( 3 ) was exploited as a starting material for the synthesis of two novel series of 5‐arylazo‐2‐hydrazonothiazoles 6a  –  6j and 2‐hydrazono[1,3,4]thiadiazoles 10a  –  10d , incorporating sulfonamide group, through its reactions with appropriate hydrazonoyl halides. The structures of the newly synthesized products were confirmed by spectral and elemental analyses. Also, the antimicrobial, anticancer, and DHFR inhibition potency for two series of thiazoles and [1,3,4]thiadiazoles were evaluated and explained by molecular docking studies and SAR analysis.     

The Effect of Stabilizing Mutations in the Central Part of α-Chain of Tropomyosin on the Bending Stiffness of Reconstructed Thin Filaments that Contain Its αβ-Heterodimers

We studied the effect of the replacement of two highly conserved noncanonical residues in the α-chain of tropomyosin, that is, Asp137 and Gly126, with the canonical residues, Leu and Arg, on the mechanical properties of reconstructed thin filaments that contain αβ-heterodimers of tropomyosin. For this purpose, the reconstructed thin filaments that contain fibrillar actin, tropomyosin, and troponin were stretched with an optical trap. The resulting strain–force diagrams were analyzed using a mathematical model proposed previously in order to estimate the bending stiffness. It was shown that the thin filaments that contain αβ-heterodimers of tropomyosin with α-chains of the pseudo-wild type, i.e., that contain the C190A substitution, have approximately the same bending stiffness as the filament with αα-homodimers of tropomyosin. The stabilizing substitution D137L in the α-chain of tropomyosin did not cause a statistically significant change in the bending stiffness of the filaments that contain αβ-heterodimers of tropomyosin, whereas the G126R and G126R/D137L substitutions led to a moderate increase in this stiffness. This increase in stiffness was, however, much less pronounced than that for the filaments that contain αα-homodimers of tropomyosin with these substitutions in both α-chains. The relationship between the results obtained in this study and the previously published data on the effects of these stabilizing substitutions in the α-chain of tropomyosin on the structural and functional properties of thin filaments with αβ-heterodimers of tropomyosin is discussed.     

Novel nucleosomal particles containing core histones and linker DNA but no histone H1

Eukaryotic chromosomal DNA is assembled into regularly spaced nucleosomes, which play a central role in gene regulation by determining accessibility of control regions. The nucleosome contains ∼147 bp of DNA wrapped ∼1.7 times around a central core histone octamer. The linker histone, H1, binds both to the nucleosome, sealing the DNA coils, and to the linker DNA between nucleosomes, directing chromatin folding. Micrococcal nuclease (MNase) digests the linker to yield the chromatosome, containing H1 and ∼160 bp, and then converts it to a core particle, containing ∼147 bp and no H1. Sequencing of nucleosomal DNA obtained after MNase digestion (MNase-seq) generates genome-wide nucleosome maps that are important for understanding gene regulation. We present an improved MNase-seq method involving simultaneous digestion with exonuclease III, which removes linker DNA. Remarkably, we discovered two novel intermediate particles containing 154 or 161 bp, corresponding to 7 bp protruding from one or both sides of the nucleosome core. These particles are detected in yeast lacking H1 and in H1-depleted mouse chromatin. They can be reconstituted in vitro using purified core histones and DNA. We propose that these ‘proto-chromatosomes’ are fundamental chromatin subunits, which include the H1 binding site and influence nucleosome spacing independently of H1.     

Calcium and EGTA Alleviate Cadmium Toxicity in Germinating Chickpea Seeds

Impact of exogenous calcium and ethylene glycol tetraacetic acid (EGTA) supplement on chickpea (Cicer arietinum L.) germinating seeds exposed to cadmium stress for 6 days was studied. Ca and EGTA late treatment (3 days) alleviated growth inhibition and decreased Cd accumulation as well as lipid peroxidation and protein carbonylation in both root and shoot cells. Exogenous effector application relieved Cd-induced cell death which was associated with a constant level of ATP, which was considered as an apoptotic-like process. Redox balance was examined through the study of the redox state of pyridine nucleotide couples NAD⁺/NADH and NADP⁺/NADPH as well as their related oxidative [NAD(P)H-oxidase] and dehydrogenase (glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and malate dehydrogenase) enzyme activities. The present research illustrated an ameliorative effect of Ca and EGTA on growth of Cd-exposed chickpea seedlings that occurs through the protection of sensitive cell sites from Cd-induced oxidation, namely membrane lipids and proteins, rather than the improvement of recycling capabilities of the cellular reducing power.     

Promotion of crown-gall tumor initiation on primary pinto bean leaves by certain inorganic salts

Crown-gall tumor initiation by Agrobacterium tumefaciens (SMITHand TOWN.) CONN, strain B6, on Phaseolus vulgaris L. var. "Pinto"was found to be sensitive to the addition of various salts tothe freshly inoculated primary leaves. Calcium, magnesium, zincand ammonium sulfates, cobalt and nickel chlorides and potassiumnitrate increased the number of tumors on inoculated leavesby 100 to 200 percent. Maximal promotions were obtained at concentrationsof 10^–7 to 10^–5 M with all of these salts exceptpotassium nitrate which was most active at 10^–2M. Sodium,potassium, calcium, magnesium and aluminum chlorides, and sodium,potassium and nickel sulfates had little or no effect on tumorinitiation. The combination of sodium sulfate with magnesiumchloride gave promotions comparable to those obtained with magnesiumsulfate, indicating that both the magnesium and sulfate ionswere necessary for the promotion obtained with this latter salt.Combining any two of the active salts at their optimum concentrationsfor promotion resulted in a reciprocal inhibition of the promotionobtained with either salt alone, suggesting these salts to beactive by different and potentially antagonistic mechanisms.The possible nature of these promotions is discussed, but theyremain too obscure to warrant a specific proposal for theirmode of action. ¹Present adress: Department of Botany, University of Khartoum,Khartoum, Sudan.     

Ultrastructural Study of the Development of Pleistophora schubergi Zwölfer, 1927 (Protozoa,Microsporida) in Larvae of the Spruce Budworm,Choristoneura fumiferana and Its Subsequent Taxonomic Change to the Genus Endoreticulatus

This study demonstrates that Pleistophora schubergi Zwölfer, 1927, a microsporidium originally isolated from the midgut epithelium of Nygmia phaeorrhoea Don (Euproctis chrysorrhoea L.) and Porthetria dispar L., and subsequently reported in several other insects including the spruce budworm, Choristoneura fumiferana (the host used in this investigation), does not belong in the genus Pleistophora Gurley, 1893. Pleistophora schubergi lacks the major features that are characteristic of Pleistophora typicalis, the type species of this genus. A comparison of ultrastructural observations reported for the type species of the genus Pleistophora, P. typicalis, and our observations of P. schubergi revealed significant differences. A thick (0.5 μm) amorphous coat, derived from parasite secretions and deposited external to the parasite plasmalemma, surrounds all developmental stages in P. typicalis. Double membranes, derived from host rough endoplasmic reticulum cisternae encircle the parasite plasmalemma of all developmental stages in P. schubergi. The sporophorous vesicle encases the spores in P. typicalis, and originates from the parasite-secreted coat that is present around meronts. In P. schubergi, the host endoplasmic reticulum cisternae form the envelope that surrounds the meronts. Moreover, the sporophorous vesicle envelope in P. typicalis persists around groups of spores, while in P. schubergi this envelope breaks easily to release the spores in the host cytoplasm. By comparing the characteristics of the microsporidium found in the spruce budworm with those of the recently created polysporous genera that sporulate within a vesicle, we found that P. schubergi does belong in the new genus Endoreticulatus Brooks et al. 1988, and consequently rename it Endoreticulatus schubergi (Zwölfer, 1927) n. comb.     

Modulation of TXA2 generation of platelets by human lipoproteins

The lipoprotein (LP) fractions VLDL, LDL, HDL2 and HDL3 were prepared by ultracentrifugation of plasma from healthy volunteers and from patients with coronary heart disease (CHD). We investigated the capacity of platelets from healthy volunteers and patients with atherosclerosis to generate thromboxane A2 (TXA2) during spontaneous clotting of whole blood under the influence of the lipoprotein fractions. In our experiments the serum concentration of TXB2, reflecting the capacity of platelets to generate TXA2 during clotting, depends on several factors: the type of LP fraction used, the blood used for generation of TXA2, and for the same LP fraction whether it was taken from plasma of healthy volunteers or patients with CHD. VLDL prepared from plasma of healthy volunteers inhibited but VLDL prepared from plasma of patients with CHD enhanced the TXA2 formation of platelets from healthy volunteers (p less than 0.05, resp.). LDL from CHD patients inhibited the TXA2 formation of platelets from atherosclerotic patients (p less than 0.01). The HDL subfractions HDL2 and HDL3 from healthy volunteers inhibited TXA2 formation by platelets from healthy volunteers as well as those from atherosclerotic patients (p less than 0.05; p less than 0.01, respectively). HDL2 from patients with CHD inhibited only the TXA2 formation of platelets from healthy volunteers (p less than 0.01), whereas HDL3 from CHD patients inhibited only the TXA2 formation of platelets from atherosclerotic patients (p less than 0.01).