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Biomedical subjects

A Zamir

Publications and source records attributed to A Zamir.

At least 19 recordsLinked to original sources

Allele frequencies of nine STR loci of Jewish and Arab populations in Israel.

DNA typing of nine short tandem repeat (STR) loci was carried out on unrelated Israeli Jewish and Arab individuals. All loci were highly polymorphic and the distribution of the obtained genotypes did not deviate from Hardy-Weinberg equilibrium. A comparison between Jewish and Arab population data revealed statistically significant differences in allele frequency distributions for some of the loci. The results presented in this study enable the use of these nine STR loci for forensic, identification and paternity cases in the Jewish and the Arab populations of Israel.

Arabs↗

Light-harvesting complex II pigments and proteins in association with Cbr, a homolog of higher-plant early light-inducible proteins in the unicellular green alga Dunaliella.

Like higher plants, unicellular green algae of the genus Dunaliella respond to light stress by enhanced de-epoxidation of violaxanthin and accumulation of Cbr, a protein homologous to early light-inducible proteins (Elips) in plants. Earlier studies indicated that Cbr was associated with the light-harvesting complex of photosystem II (LHCII) and suggested it acted as a zeaxanthin-binding protein and fulfilled a photo-protective function (Levy et al. 1993, J. Biol. Chem. 268: 20892-20896). To characterize the protein-pigment subcomplexes containing Cbr in greater detail than attained so far, thylakoid membranes from Dunaliella salina grown in high light or normal light were solubilized with dodecyl maltoside and fractionated by isoelectric-focusing. Analysis of the resolved LHCII subcomplexes indicated preferred associations among the four LHCIIb polypeptides and between them and Cbr: subcomplexes including Cbr contained one or two of the more acidic of the four LHCIIb polypeptides as well as large amounts of lutein and zeaxanthin relative to chlorophyll a/b. After sucrose gradient centrifugation, Cbr free of LHCIIb polypeptides was detected together with released pigments; this Cbr possibly originated in subcomplexes dissociated in the course of the analysis. These results agree with the conclusion that Cbr is part of the network of LHCIIb protein-pigment complexes and suggest that the role played by Cbr involves the organization and/or stabilization of assemblies highly enriched in zeaxanthin and lutein. Such assemblies may function to protect PSII from photodamage due to overexcitation.

Centrifugation, Density Gradient↗

An evaluation of the relevance of routine DNA typing of fingernail clippings for forensic casework.

DNA extracted from fingernail clippings of victims in forensic cases is a possible source of DNA from the perpetrator in cases where victims struggled or defended themselves. The source of this DNA on a victim's fingernails could possibly originate from contact with the suspect's blood, saliva, semen or scratched skin. In this technical note we evaluate the relevance of routine DNA typing of fingernail clippings in the forensic biology laboratory when, in real casework, normally only small quantities of nail material is sent. This was carried out by extracting DNA from fingernail clippings from a number of volunteers, before and after aggressively scratching other volunteers. No blood was drawn from the scratching, but skin flakes were observed under the nails before cutting and subsequent DNA typing. The DNA extracted was then typed using the STR systems: HUMTHO1, HUMTPOX and HUMCSF1PO (CTT triplex) and the system of D1S80. These profiles were compared with profiles achieved by similar typing of buccal swabs as a reference from each volunteer. In this study, the profile detected from each volunteer's clippings was the same before and after scratching, and matched the profile of the corresponding volunteer as defined by typing each volunteer's reference buccal swab. Fingernail clippings that are sent to our lab in actual casework are usually so small that additional treatment by swabbing or removing debris from below the clipping is not possible. For this reason, in this simulation the entire clippings were used for DNA extraction, to maximize the possibility of finding an additional profile. In conclusion, the findings from this study show that although the profiles obtained when typing fingernail clippings are those of the donors themselves, we suggest that typing of fingernail clippings should be carried out in forensic cases only when relevant. We would suggest that fingernail clippings not be routinely sent to the biology laboratory as items of evidence to be tested.

Forensic Medicine↗

Threat mail and forensic science: DNA profiling from items of evidence after treatment with DFO.

Two cases of threatening letters with their accompanying envelopes were received to the Division of Forensic Identification unit of the Israel Police. The envelopes, including the stamps, and the letters were initially examined for latent fingerprints by the DFO reagent, known to cause degradation of DNA. Although no latent fingerprints could be visualized on any of the items, the biology laboratory using organic DNA extraction, was successful in defining genetic profiles from all the items employing six STR loci, even after treatment with DFO. In a controlled experiment, a known donor attached a stamp, by licking, to an envelope. This item was treated with DFO and then profiled using STR loci. The results showed that previous DFO treatment on the control stamp before DNA analysis had no negative effects on obtaining the DNA profile of the known donor using STR loci.

Aza Compounds↗

Fingerprints and DNA: STR typing of DNA extracted from adhesive tape after processing for fingerprints.

An exhibit that is often received for examination in cases of robbery or terrorist activity is adhesive tape. This type of exhibit can often, but not always, be successfully processed for fingerprints. The question arises whether or not it is possible to extract and type DNA after the tape has been sequentially processed for fingerprints. In this work, various donors left fingerprints on the adhesive side of tapes. The tapes were then sequentially processed for fingerprints using an alternate light source, cyanoacrylate fuming, and staining with BY-40 and then crystal violet. DNA was subsequently successfully extracted, amplified and typed for six STR loci.

Adhesives↗

Differential responses to different light spectral ranges of violaxanthin de-epoxidation and accumulation of Cbr, an algal homologue of plant early light inducible proteins, in two strains of Dunaliella.

Unicellular green algae of the genus Dunaliella, similar to higher plants, respond to light stress by enhanced de-epoxidation of violaxanthin and accumulation of Cbr, a protein homologous to early light inducible proteins (Elips) in plants. These proteins belong to the superfamily of chlorophyll a/b binding proteins. Two Dunaliella strains, D. bardawil and D. salina, were compared for these two responses under light in the UVA, blue, green and red spectral ranges. In D. bardawil, the two stress responses were similarly induced under UVA, blue or red light and to a lesser extent under green light. In D. salina, a similar spectral range dependence was exhibited for violaxanthin de-epoxidation. However, Cbr accumulated only under UVA or blue light but not under green or red light. A strong synergistic effect of a low dose of blue light superimposed on red light resulted in Cbr accumulation. These results reveal strain-specific differences in spectral range requirements of the two light-stress responses. In the two strains, violaxanthin de-epoxidation is triggered under photosynthetically-active spectral ranges but at least in D. salina, Cbr accumulation appears to require a specific light signal additionally to a signal(s) generated by light stress.

Journal Article↗

Iron uptake by the halotolerant alga Dunaliella is mediated by a plasma membrane transferrin.

A 150-kDa transferrin-like protein (Ttf) is associated with the plasma membrane of the halotolerant unicellular alga Dunaliella salina (Fisher, M., Gokhman, I., Pick, U., and Zamir, A. (1997) J. Biol. Chem. 272, 1565-1570). The Ttf level rises with medium salinity or upon iron depletion. Evidence that Ttf is involved in iron uptake by Dunaliella is presented here. Algal iron uptake exhibits characteristics resembling those of animal transferrins: high specificity and affinity for Fe3+ ions, strict dependence on carbonate/bicarbonate ions, and very low activity in acidic pH. Reducing the level of Ttf by mild proteolysis of whole cells is accompanied by lowered uptake activity. Conversely, accumulation of high levels of Ttf is correlated with an enhancement of iron uptake. Kinetically, iron uptake consists of two steps: an energy-independent binding of iron to the cell surface and an energy-dependent internalization. Salinities as high as 3.5 M NaCl do not inhibit iron uptake or decrease the apparent affinity for Fe3+ ions, implying that Ttf activity is not affected by high salt. These results indicate that transferrins, hitherto identified only in animals, are present and function in iron transport also in plant systems.

Cell Membrane↗

Immunoproliferative small intestinal disease in a 16-year-old boy presenting as severe malabsorption with excellent response to tetracycline treatment.

Immunoproliferative small intestinal disease (IPSID) is a rare lympho-proliferative disorder of the upper small intestine. It is considered a special form of MALT lymphoma with propensity to malignant transformation. This disorder is rare in pediatric literature. We report a case of IPSID in a 16-year-old boy with low-grade malignant transformation, presenting as severe malnutrition and a possible association with Helicobacter pylori. The patient responded well to an extended treatment with tetracycline and eradication of H. pylori.

Adolescent↗

A structurally novel transferrin-like protein accumulates in the plasma membrane of the unicellular green alga Dunaliella salina grown in high salinities.

The alga Dunaliella salina is outstanding is its ability to withstand extremely high salinities. To uncover mechanisms underlying salt tolerance, a search was carried out for salt-induced proteins. The level of a plasma membrane 150-kDa protein, p150, was found to increase with rising external salinity (Sadka, A., Himmelhoch, S., and Zamir, A. (1991) Plant Physiol. 95, 822-831). Based on its cDNA-deduced sequence, p150 belongs to the transferrin family of proteins so far identified only in animals. This, to our best knowledge, is the first demonstration of a transferrin-like protein in a photosynthetic organism. Unlike animal transferrins, p150 contains three, rather than two, internal repeats and a COOH-terminal extension including an acidic amino acid cluster. In intact cells p150 is degraded by Pronase, indicating that the protein is extracellularly exposed. The relationship of p150 to iron uptake is supported by the induction of the protein in iron-deficient media and by its radioactive labeling in cells grown with 59Fe. Accumulation of p150 is transcriptionally regulated. It is proposed that p150 acts in iron uptake other than by receptor-mediated endocytosis and that its induction permits the cells to overcome a possible limitation in iron availability under high salinities.

Algal Proteins↗

A salt-resistant plasma membrane carbonic anhydrase is induced by salt in Dunaliella salina.

The mechanisms allowing proliferation of the unicellular green alga Dunaliella salina in up to saturating NaCl concentrations are only partially understood at present. Previously, the level of a plasma membrane Mr 60,000 protein, p60, was found to increase with rising external salinities. Based on cDNA cloning and enzymatic assays, it is now shown that p60 is an internally duplicated carbonic anhydrase, with each repeat homologous to animal and Chlamydomonas reinhardtii carbonic anhydrases, but exceptional in the excess of acidic over basic residues. Increasing salinities, alkaline shift, or removal of bicarbonate induced in D. salina parallel increases in the levels of p60, its mRNA, and external carbonic anhydrase activity. Moreover, purified p60 exhibited carbonic anhydrase activity comparable to other carbonic anhydrases. A p60-enriched soluble preparation showed maximal carbonic anhydrase activity at approximately 1.0 M NaCl and retained considerable activity at higher salt concentrations. In contrast, a similar preparation from C. reinhardtii was approximately 90% inhibited in 0.6 M NaCl. These results identified p60 as a structurally novel carbonic anhydrase transcriptionally regulated by CO2 availability and exhibiting halophilic-like characteristics. This enzyme is potentially suited to optimize CO2 uptake by cells growing in hypersaline media.

Alkalies↗

Chaperonins as potential gene regulatory factors. In vitro interaction and solubilization of NifA, the nif transcriptional activator, with GroEL.

A previous study (Govezensky, D., Greener, T., and Zamir, A. (1991) J. Bacteriol. 20, 6339-6346) indicated that the chaperonin GroEL was required for maximal expression from nif promoters in Klebsiella pneumoniae and nif-transformed Escherichia coli. That this requirement stemmed from the ability of GroEL to properly fold NifA, the nif transcriptional activator, was first supported by co-immunoprecipitation of NifA in K. pneumoniae extracts with anti-GroEL antibodies. In the present in vitro study, NifA, partially purified from E. coli overexpressing the protein, was diluted from a 6 M urea solution into a refolding buffer in the presence or absence of GroEL. Dilution in the absence of GroEL caused the complete precipitation of NifA. When present in the dilution buffer, GroEL bound NifA and maintained it in a soluble state. GroEL was also found to bind NifA newly synthesized in an in vitro translation system. For both NifA preparations, cochaperonin GroES and ATP promoted release of NifA from GroEL. These results provide evidence for the association of NifA with GroEL and for the role of both GroEL and GroES in the solubilization and thereby folding of the nif transcriptional activator.

Bacterial Proteins↗

Wall hangings, hairs and the Holocaust.

Two wall hangings, from the time of the Second World War, were reportedly made from human hair. Laboratory examination of the hangings confirmed this fact. Fibers, holding the wall hangings together, and the weave of the hangings were also examined. Results and implications of the findings are discussed.

Animals↗

Cbr, an algal homolog of plant early light-induced proteins, is a putative zeaxanthin binding protein.

The cbr gene, previously cloned from the unicellular green alga Dunaliella bardawil, is transcriptionally and translationally activated in parallel to accelerated carotenogenesis in response to light stress conditions. The product of cbr, structurally similar to Elips (early light-induced proteins of higher plants), is associated with a minor light harvesting complexes of photosystem II component (Levy, H., Gokhman, I., and Zamir, A. (1992) J. Biol. Chem. 267, 18831-18836). This study examines the relationship between the induction of Cbr and another plant response to light stress, the deepoxidation of violaxanthin to zeaxanthin. A parallel between the two processes was observed in cells exposed to high light, starved for sulfate, or treated with norflurazon, a herbicide inducing photooxidative damage by inhibiting de novo carotenoid biosynthesis. When highly illuminated cells were returned to normal light, Cbr decayed in parallel to the reepoxidation of zeaxanthin to violaxanthin. Evidence for the physical association of Cbr and zeaxanthin was provided by nondenaturing gel electrophoresis. In cells transferred from low to high light, zeaxanthin was associated with the faster migrating of two electrophoretically resolved fractions of light harvesting complexes of photosystem II that also contained Cbr. In cells growing under normal light, violaxanthin was bound equally to the two fractions. Based on these results we propose that Cbr/early light-induced proteins bind zeaxanthin to form photoprotective complexes within the light-harvesting antennae.

Carotenoids↗

A novel multicopy suppressor of a groEL mutation includes two nested open reading frames transcribed from different promoters.

When present on a multicopy plasmid, a newly discovered gene (sugE) mapping to 94 min on the Escherichia coli chromosome, suppresses a groEL mutation and mimics the effects of groE overexpression. A groEL mutant of E.coli, transformed with the Klebsiella pneumoniae nif gene cluster, failed to accumulate nitrogenase components [Govezensky et al. (1991) J. Bacteriol., 173, 6339-6346]. Transformation with sugE reversed the mutant phenotype. In wild type K.pneumoniae, transformation with sugE accelerated the rate of nitrogenase biogenesis after nif derepression. In E.coli, transformation with sugE enabled bacteriophage T4 growth in a groEL mutant. A continuous 178 codon open reading frame (ORF) in sugE encloses another, in-frame, 105 codon ORF similar to a predicted ORF in Proteus vulgaris. In vivo products of both sugE ORFs were observed in transformants expressing the gene from a T7 promoter. In non-transformed cells, a typical sigma 70-dependent promoter found upstream of the larger ORF directs sugE transcription during growth at 30 degrees C. At elevated temperatures or in stationary phase cells, another promoter, found within the coding sequence upstream of the smaller ORF, is activated independently of sigma 32. The results suggest that sugE encodes a chaperonin-related system whose composition might vary with temperature and growth phase.

Amino Acid Sequence↗

Regulation and light-harvesting complex II association of a Dunaliella protein homologous to early light-induced proteins in higher plants.

The unicellular green alga Dunaliella bardawil responds to high light, nutrient deprivation, and several other types of stress by massive accumulation of beta-carotene. We have previously cloned a nuclear gene, cbr, that is co-induced with accelerated carotenogenesis. The predicted product of cbr is closely related to early light-induced proteins (Elips) of higher plants, and also shows resemblance to chlorophyll a/b-binding proteins. In the present study, the determination of the cbr transcription start site supported the previously proposed site of translation initiation. Antibodies raised against a synthetic oligopeptide matching the predicted sequence of Cbr recognized two polypeptides of apparently 17 and 19 kDa that were induced in parallel to cbr transcript accumulation in highly illuminated or sulfate-starved D. bardawil cells. The antibodies also cross-reacted with an approximately 20-kDa polypeptide in sulfate-starved Dunaliella salina. In both D. bardawil and D. salina, Cbr was found exclusively in the thylakoid membranes. After mild solubilization, Cbr co-fractionated with light-harvesting complex II (LHCII) in sucrose gradient centrifugation and gel electrophoresis and was specifically associated with a minor LHCII complex. An occasionally observed, faster mobility Cbr form, free of LHCII, was probably released from the larger complex. These results support the conclusion that Cbr belongs to a class of stress-induced proteins transiently associated with antennae complexes.

Amino Acid Sequence↗

Co-regulation of a gene homologous to early light-induced genes in higher plants and beta-carotene biosynthesis in the alga Dunaliella bardawil.

Dunaliella bardawil, a unicellular green alga that can be induced to accumulate massive amounts of beta-carotene, is particularly suitable for studies of carotenogenesis regulation and its links to developmental and adaptive processes in the chloroplast. A cDNA clone corresponding to a transcript accumulating coordinately with carotenogenesis induction was isolated by differential hybridization of a cDNA library made from intensely illuminated cells. This transcript was also abundant in algal mutants able to accumulate beta-carotene under relatively low light intensity. DNA sequence analysis indicates that cbr (for carotene biosynthesis-related) is closely related to early light-induced genes (elip) of higher plants. Similarity also exists between repeated oligopeptide motifs in Cbr and Cab proteins of photosystems I and II. Three upstream direct repeats in cbr include an octamer and hexamer related to mammalian sterol regulatory elements. The relationship between cbr transcript and beta-carotene accumulation, the structural similarity between Cbr and Cab proteins, and the presence of potential SRE-1 elements lead us to propose that Cbr represents novel carotenoid binding proteins, whose synthesis might be coordinated with carotenogenic enzymes via an evolutionary conserved regulatory mechanism.

Amino Acid Sequence↗