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PubMed1989

Liver uricase in Camelus dromedarius: purification and properties.

1. Uricase (urate: oxygen oxidoreductase, EC 1.7.3.3) was purified 750-fold from the liver of Camelus dromedarius. 2. The enzyme is a tetramer with a Mr of 100,000, displays high specificity for uric acid with a Km of 12 microM and is inhibited by a selected number of purine derivatives carrying oxygen at the C2 position. 3. The effect of pH and the inhibition by thiol compounds and chelating agents on the enzyme activity is reported. 4. Some lines of evidence suggesting the possibility of interaction of camel liver uricase with oligonucleotides are presented.

Animals↗
PubMed1990

A comparison of cardiac glutathione S-transferases from wild and domestic animals.

1. Cardiac glutathione S-transferases from wild animals; hyena, red fox, porcupine, coypu and mountain gazelle were purified and compared with the enzymes from domestic animals; cow, camel, goat and sheep. 2. By using 1-chloro-2,4-dinitrobenzene as a substrate, domestic hearts expressed higher glutathione conjugating activity than wild animals hearts. 3. In all the studied hearts, the bulk of the activity was associated with near neutral and acidic glutathione S-transferase isozymes with pI values ranging from 4 to 7.4. 4. The enzymes from domestic animals displayed homodimeric structure of 25,000 mol. wt subunit while of the wild animals both hyena and coypu displayed homodimers of 26,500 mol. wt subunit and the rest exhibited heterodimers of 25,000 and 28,000 mol. wt subunits.

Animals↗
PubMed1995

Characterization of a GlyCAM1-like gene (glycosylation-dependent cell adhesion molecule 1) which is highly and specifically expressed in the lactating bovine mammary gland.

A bovine cDNA library, derived from the mammary gland of a lactating cow, was screened for identifying transcripts that specifically occur during lactation by means of differential hybridisation. Several of the clones isolated by this procedure shared 55 and 57% similarity with the mouse and rat glycosylation-dependent cell adhesion molecule 1 (GlyCAM1) cDNAs, respectively. Although the mouse and cattle proteins showed an overall similarity of only 41%, two specific regions of the proteins showed 83 and 81% similarity, respectively. The bovine protein also showed 55% similarity with a small protein isolated from the whey fraction of camel milk. Northern blot analysis showed that high-level expression of this gene was only observed in the mammary gland of lactating cows. The complete gene was isolated from a bovine genomic library and its organisation was determined. The gene was 2.5 kb in length and split into four exons. The size and organization of the gene as well as the position of the introns was identical to that of the mouse GlyCAM1 gene. In accordance with the tissue-specific expression of this gene in the mammary gland of lactating animals, potential mammary gland factor (MGF) binding sites were present in the promoter region of the gene. Based on the data presented in this study it is highly likely that this gene is the bovine homologue to the rat and mouse GlyCAM1 genes.

Amino Acid Sequence↗
PubMed1987

Effects of Org OD 14 on pituitary and peripheral beta-endorphin in castrated rats and post-menopausal women.

The aim of the first part of this study was to evaluate the effects of a new synthetic steroid (7 alpha,17 alpha)-17-hydroxy-7-methyl-19-norpregn-5(10)-en-20-yn-3-one (Org OD 14), on anterior pituitary (AP) and neurointermediate pituitary lobe (NIL) contents and on circulating levels of beta-endorphin (beta-EP) in rats. Three weeks after ovariectomy, groups of 9 rats were treated with either Org OD 14 (2 or 10 micrograms/day/rat for 14 days) or a placebo. In addition, 2 groups of ovariectomized rats were also treated with oestradiol benzoate (EB) (2 or 10 micrograms/day/rat for 14 days) to compare the effectiveness of the new steroid with that of a classical oestrogenic substance. beta-Ep concentrations were measured in plasma and in AP and NIL extracts by means of double-antibody radioimmunoassay (RIA), employing a specific anti-camel beta-EP (C-terminal fragment). Both doses of Org OD 14 induced a significant dose-related increase in plasma and pituitary lobe beta-EP concentrations as compared with the results on placebo treatment. By comparison, EB was active only at a dose of 10 micrograms/day. Despite the common stimulatory effects of EB and Org OD 14 on pituitary beta-EP, these findings suggest that the two steroids have different modes of action. The second part of the study investigated the changes in beta-EP and beta-lipotrophin (beta-LPH) plasma levels in a group of post-menopausal women treated for 6 months with Org OD 14 (2.5 mg/day) in comparison with the levels in a placebo-treated group. The clinical efficacy of Org OD 14 treatment in post-menopausal symptoms was confirmed, as well as its lack of or only transient effect on plasma lipids and lipoproteins. beta-EP and beta-LPH plasma levels were significantly higher in the Org OD 14-treated group than in the placebo group as from the second month until the end of the observation period.

Adult↗
PubMed2004

Cystic echinococcosis in Argentina: evolution of metacestode and clinical expression in various Echinococcus granulosus strains.

Echinococcus granulosus hydatid cysts were examined in 41 patients from Neuquén and Tucumán provinces in Argentina. Sequencing of the mitochondrial cytochrome c oxidase subunit 1 (CO1) revealed in 19 patients common sheep strain (G1), in 6 patients Tasmania sheep strain (G2), in 1 patient cattle strain (G5), and in 15 patients camel strain (G6). In Argentina the only known is the domestic cycle that affects dogs and herbivorous, including ovine, swine, cattle and goats. These strains produced a total of 58.6% of primary liver infections, 29.2% primary in lung, 2.4% primary in spleen and 9.8% were multiorgan abdominal infections. The metacestode was classified using the evolutive stages proposed by WHO-IWGE (from CE1 to CE5). We estimated that CE1 cyst has a duration of about 22 years, CE2 of 14 years, CE3 of 10 years, CE4 of 19 years and CE5 was not determined. The active types CE1 and CE2 reached 75% of all cases from all strains. In 36 patients with cysts from G1, G5 and G6 strain, there were only two asymptomatic cases. The strains of the E. granulosus complex do not present important clinical differences; only G6 seems to have higher growth rate.

Adolescent↗
PubMed2005

Morbidity and nutrition patterns of three nomadic pastoralist communities of Chad.

As a part of an interdisciplinary research and action programme, morbidity and nutritional patterns were assessed in three nomadic communities: Fulani and Arab cattle breeders and Arab camel breeders, of two prefectures in Chad. The predominant morbidity pattern of Chadian nomadic pastoralists (representing approximately 10% of the total population of the country) had not been documented so far. A total of 1092 women, men and children was examined by a physician and interviewed during two surveys in the dry season and one in the wet season (1999--2000). Participants with no complaint were rare. Pulmonary disorders (e.g. bronchitis) were most often diagnosed for children under 5 years of age. Of the adult participants, 4.6% were suspected of tuberculosis. Febrile diarrhoea occurred more often during the wet season when access to clean drinking water was precarious. Malaria was only rarely clinically diagnosed among Arabs during the dry season, whereas Fulani, who stayed in the vicinity of Lake Chad, were also affected during this period. A 24-h dietary recall showed that less Arab women than men consumed milk during the dry season (66% versus 92%). Malnutrition was only documented for 3 out of 328 children (0--14 years). Arab women in childbearing age had a higher proportion of children not surviving when compared to Fulani women (0.2 versus 0.07). This study identified several implications for reseach and interventions in nomadic settings. Innovative and integrated health services for nomads can possibly be extended to many settings as nomadic pastoralists have in common a similar way of life driven by the needs of their animals.

Adolescent↗
PubMed2006

A myth too tough to die: the dead of disasters cause epidemics of disease.

Myths abound in the practice of health care, death, and disease. Akin to the old adage of swallowing camels and straining gnats, the myth that mass fatalities cause epidemics of disease following natural or other disasters is alive and well. Despite the findings of observers, microbiologists, epidemiologists, and other scientists, even medical doctors and public health professionals lend support to the ancient belief and rush into mass graves or mass cremations the bodies of those victims of trauma in a disaster. Putting this myth to rest depends on use of information concerning the transmission of the organisms that cause disease, the sources of those organisms, and the hosts or suspected hosts that will be receptive to those organisms to result in disease. The information provided by recent investigators of disasters in the region of the Americas and comments from those who have reviewed the literature on the subject of the myth concerning the dead and epidemics following disasters have provided the basis for some concise guidelines for placing this myth in the archives of other traditions without foundation. Education of the public and the news media are the responsibility of those who are aware of the fallacies in this belief to bring about the demise of this myth.

Cadaver↗
PubMed2006

Cigarette promotional offers: who takes advantage?

BACKGROUND: Promotional offers on cigarettes (e.g., dollar-off, multipack discounts) composed the largest share of tobacco industry marketing expenditures, totaling $8.9 billion, or 72% of the total budget in 2002. Internal industry documents indicate that young adults, potential quitters, and other price-sensitive groups are the targets of these marketing tactics. How effective they are in actually reaching these groups in the general population of smokers has not yet been investigated. METHODS: Data were from 4618 current smokers responding to the large, random-digit-dialed population-based 2002 California Tobacco Survey. The characteristics were identified of smokers who reported that they used these offers "every time I see one." RESULTS: Thirty-five percent of smokers used promotional offers every time they saw one. Multivariate analyses identified young adults, women, African Americans, those with higher daily cigarette consumption, and those worried about cigarette costs as more likely to use promotional offers at every opportunity. Smokers most committed to quitting were no more likely to use promotional offers than those with no intention to quit. Cigarette brand was highly correlated with age and race/ethnicity, and therefore was not included in the multivariate analysis. Those who smoked menthol cigarettes and Camels, more often young adults and African Americans, were much more likely than those of other brands to use promotional offers. CONCLUSIONS: With the exception of smokers intending to quit, cigarette promotional offers are effectively reaching most industry-targeted groups. Importantly, young adults, who have the greatest long-term customer potential, are responding.

Adolescent↗
PubMed2006

Effect of follicular size on in vitro developmental competence of oocytes and viability of embryos after transfer in the dromedary (Camelus dromedarius).

The aim of this work was to determine the effect of follicle size on camel oocyte quality as measured by developmental competence in vitro and in vivo. Ovaries from a local slaughterhouse were dissected to obtain two classes of follicle size: small (3-6 mm) and large (>6 mm) follicles. Quality of the oocytes was assessed after in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC) of cumulus oocyte complexes (COCs). All cultures were done in four replicates at 38.5 degrees C, under 5% CO(2) and high humidity (>95%). Only COCs with cumulus and homogenous (dark) cytoplasm were used. The COCs were matured for 28 h in TCM-199 medium supplemented with 10% heat-treated fetal calf serum (FCS), 10 ng/mL EGF, and 250 microM cysteamine. Nuclear maturation rate for each class of follicle size was determined by contrast phase microscopy in a sample of COCs (n=30) denuded, fixed and stained with aceto-orcein. In vitro fertilization was performed using fresh semen (0.5 x 10(6)spermatozoa/mL in modified TALP-solution). Fertilized oocytes were cultured in mKSOMaa, under 5% O(2) and 90% N(2). The percentage of COCs reaching metaphase II (MII) after 28 h of maturation was 87% (26/30) and 73% (22/30) for oocytes originating from large and small follicles, respectively (P>0.1). The rate of total cleavage (two cells to blastocyst stage) was greater (P<0.05) for oocytes originating from large follicles (72%; 116/162) than for those derived from small follicles (59%; 140/237). The percentage of fertilized oocytes reaching the blastocyst stage was 35% (57/162) and 20% (48/237) for oocytes collected from large and small follicles, respectively (P<0.05). The viability of in vitro-produced hatched blastocyst from the two groups (15 from 3 to 6mm follicle size and 22 from follicles >6 mm) was assessed by transfer to synchronized recipients. None of the hatched blastocysts from small follicles resulted in a pregnancy whereas 68% (15/22) of the transferred hatched embryos from large follicles developed into a 25-day pregnancy. Of the resulting 15 pregnancies, 53% (n=8) aborted (five between 2 and 4 months and three between 5 and 7 months of pregnancy). The remaining seven pregnant females gave birth to normal healthy offsprings (four females and three males). The present study shows that dromedary oocytes developmental competence is acquired late during the final phase of follicular development and this developmental ability translates into greater pregnancy rates after transfer of in vitro produced hatched blastocysts.

Animals↗
PubMed2004

Cloning and characterization of 5' upstream promoter region of rat WAP gene.

Regulatory regions of genes encoding milk proteins are frequently used to produce in the mammary gland of transgenic animals a variety of pharmaceutically and medically important human proteins. One such example is the whey acidic protein (WAP) promoter region, identified so far in the genome of mouse, rat, rabbit, camel, pig, brushtail possum and Tammar wallaby. The aim of the present study was cloning and characterization of the 5' upstream promoter region of rat WAP gene. Using Genome Walking procedure, we cloned the region extending from -849 to -3671 bp. We have shown that there are two conserved regions highly similar to hypersensitive sites present in mouse and rabbit upstream region of WAP gene with binding sites for STAT5 transcription factor, essential for expression of WAP gene in mammary glands during lactation. We characterized dispersed and tandem repeats in the upstream region of rat WAP gen localized not far away from the translation initiation site.

5' Flanking Region↗
PubMed2005

Position-specific incorporation of a highly photodurable and blue-laser excitable fluorescent amino acid into proteins for fluorescence sensing.

A new fluorescent amino acid, L-2-acridonylalanine, was incorporated into proteins at specific positions using 4-base codon/anticodon strategy. The efficiency of the incorporation was high enough to obtain enough quantities of the mutants. The acridonyl group was highly fluorescent when it was excited at the wavelengths of blue-lasers and was highly photodurable compared with conventional fluorophores often used for biological analyses. The fluorescence intensity was sensitive to small changes in the polarity of the environment. When the nonnatural amino acid was incorporated into specific positions of streptavidin, the mutant protein worked as a fluorescent sensor to biotin. Similarly, when the amino acid was incorporated into camel single-chain antibody, the mutant protein sensitively responded to the antigen molecule. The high incorporation efficiency, the high photodurability, the excitability with blue-lasers, and high sensitivity to the environment make the acridonylalanine as the promising fluorescent amino acid for sensing small molecules when incorporated into specific positions of various antibodies, receptors, and enzymes.

Acridines↗
PubMed2005

Thermonuclease gene as a target for specific identification of Staphylococcus intermedius isolates: use of a PCR-DNA enzyme immunoassay.

A PCR-DNA enzyme immunoassay (PCR-DEIA) was developed for identification of the coagulase-positive species Staphylococcus intermedius. Two PCR primers and a hybridization probe were designed to target specific sequences of the S. intermedius thermonuclease (nuc) gene. In addition to S. intermedius reference strains, the PCR-DEIA was tested using 295 veterinary and human S. intermedius isolates. A specific 933-bp DNA fragment was successfully amplified in 281 (94.9%) S. intermedius isolates. Five canine isolates showed an unexpected 2.8-kbp band. Except for 10 amplicons derived from equine, camel, and pigeon isolates, all positive PCR results (n = 288, 96.6%) were confirmed by the colorimetric microtiter plate DEIA hybridization. Isolates that failed both in amplification and DEIA hybridization were only observed in equine isolates (10/23, 43.5%). Except for the limitations with isolates of hoofed animals, the S. intermediusnuc PCR assay has potential for rapid identification of S. intermedius and differentiation from other coagulase-positive staphylococci including S. aureus.

Animals↗
PubMed2005

Leptin expression in ruminants: nutritional and physiological regulations in relation with energy metabolism.

Leptin, mainly produced in adipose tissue (AT), is a protein involved in the central and/or peripheral regulation of body homeostasis, energy intake, storage and expenditure, fertility and immune functions. Its role is well documented in rodent and human species, but less in ruminants. This review is focused on some intrinsic and extrinsic factors which regulate adipose tissue leptin gene expression and leptinemia in cattle, sheep, goat and camel: age, physiological status (particularly pregnancy and lactation) in interaction with long-term (adiposity) and short-term effects of feeding level, energy intake and balance, diet composition, specific nutrients and hormones (insulin, glucose and fatty acids), and seasonal non-dietary factors such as photoperiod. Body fatness strongly regulates leptin and its responses to other factors. For example, leptinemia is higher after underfeeding or during lactation in fat than in lean animals. Physiological status per se also modulates leptin expression, with lactation down-regulating leptinemia, even when energy balance (EB) is positive. These results suggest that leptin could be a link between nutritional history and physiological regulations, which integrates the animal's requirements (e.g., for a pregnancy-lactation cycle), predictable food availability (e.g., due to seasonal variations) and potential for survival (e.g., body fatness level). Reaching permissive leptin thresholds should be necessary for pubertal or postpartum reproductive activity. In addition to the understanding of leptin yield regulation, these data are helpful to understand the physiological significance of changes in leptin secretion and leptin effects, and how husbandry strategies could integrate the adaptative capacities of ruminant species to their environment.

Adipose Tissue↗
PubMed2005

Expression of leptin and its receptors in various tissues of ruminants.

The energy metabolism of domestic animals is under the control of hormonal factors, which include thyroid hormones and leptin. Leptin signals from the periphery to the centre. It is mostly produced in the white adipose tissue and informs the central nervous system (CNS) about the total fat depot of the body. Low and high levels of leptin induce anabolic and catabolic processes, respectively. Besides controlling the food uptake and energy expenditure leptin is also involved in regulation of the reproduction and the immune system. Leptin is produced in several tissues other than fat. In the present paper the leptin expression of ruminant (Egyptian water buffalo, cow, and one-humped camel) tissues are examined. The mammary gland produces leptin in each species investigated. The local hormone production contributes to milk leptin and most probably helps to maintain lactation. Considerable leptin receptor expression was observed in the milk-producing epithelial cells, which is the same cell type that produces most of the udder leptin. Based on the results tissues participating in production have an autoregulative mechanism through which tissues can be relatively independent of the plasma leptin levels in order to maintain the desired function.

Animals↗
PubMed2006

Ruminants genome no longer contains Whey Acidic Protein gene but only a pseudogene.

Whey Acidic Protein (WAP) has been identified in the milk of only a few species, including mouse, rat, rabbit, camel, pig, tammar wallaby, brushtail possum, echidna and platypus. Despite intensive studies, it has not yet been found in the milk of Ruminants. We have isolated and characterized genomic WAP clones from ewe, goat and cow, identified their chromosomal localization and examined the expression of the endogenous WAP sequence in the mammary glands of all three species. The WAP sequences were localized on chromosome 4 (4q26) as expected from comparative mapping data. The three ruminant WAP sequences reveal the same deletion of a nucleotide at the end of the first exon when compared with the pig sequence. Due to this frameshift mutation, the putative proteins encoded by these sequences do not harbor the features of a usual WAP protein with two four-disulfide core domains. Moreover, RT-PCR experiments have shown that these sequences are not transcribed and are, thus, pseudogenes. This loss of functionality of the gene in Ruminants raises the question of the biological role of the WAP. Some putative roles previously suggested for WAP are discussed.

Animals↗
PubMed2005

Acrolein in cigarette smoke inhibits T-cell responses.

BACKGROUND: Cigarette smoking inhibits T-cell responses in the lungs, but the immunosuppressive compounds have not been fully identified. Cigarette smoke extracts inhibit IL-2, IFN-gamma, and TNF-alpha production in stimulated lymphocytes obtained from peripheral blood, even when the extracts were diluted 100-fold to 1000-fold. OBJECTIVE: The objective of these studies was to identify the immunosuppressive compounds found in cigarette smoke. METHODS: Gas chromatography/mass spectroscopy and HPLC were used to identify and quantitate volatile compounds found in cigarette smoke extracts. Bioactivity was measured by viability and production of cytokine mRNA and protein levels in treated human lymphocytes. RESULTS: The vapor phase of the cigarette smoke extract inhibited cytokine production, indicating that the immunosuppressive compounds were volatile. Among the volatile compounds identified in cigarette smoke extracts, only the alpha,beta-unsaturated aldehydes, acrolein (inhibitory concentration of 50% [IC50] = 3 micromol/L) and crotonaldehyde (IC50 = 6 micromol/L), exhibited significant inhibition of cytokine production. Although the levels of aldehydes varied 10-fold between high-tar (Camel) and ultralow-tar (Carlton) extracts, even ultralow-tar cigarettes produced sufficient levels of acrolein (34 micromol/L) to suppress cytokine production by >95%. We determined that the cigarette smoke extract inhibited transcription of cytokine genes. The inhibitory effects of acrolein could be blocked with the thiol compound N-acetylcysteine. CONCLUSION: The vapor phase from cigarette smoke extracts potently suppresses cytokine production. The compound responsible for this inhibition appears to be acrolein.

Acetylcysteine↗
PubMed2004

Crystal structure of HEL4, a soluble, refoldable human V(H) single domain with a germ-line scaffold.

The antigen binding site of antibodies usually comprises associated heavy (V(H)) and light (V(L)) chain variable domains, but in camels and llamas, the binding site frequently comprises the heavy chain variable domain only (referred to as V(HH)). In contrast to reported human V(H) domains, V(HH) domains are well expressed from bacteria and yeast, are readily purified in soluble form and refold reversibly after heat-denaturation. These desirable properties have been attributed to highly conserved substitutions of the hydrophobic residues of V(H) domains, which normally interact with complementary V(L) domains. Here, we describe the discovery and characterisation of an isolated human V(H) domain (HEL4) with properties similar to those of V(HH) domains. HEL4 is highly soluble at concentrations of > or =3 mM, essentially monomeric and resistant to aggregation upon thermodenaturation at concentrations as high as 56 microM. However, in contrast to V(HH) domains, the hydrophobic framework residues of the V(H):V(L) interface are maintained and the only sequence changes from the corresponding human germ-line segment (V3-23/DP-47) are located in the loops comprising the complementarity determining regions (CDRs). The crystallographic structure of HEL4 reveals an unusual feature; the side-chain of a framework residue (Trp47) is flipped into a cavity formed by Gly35 of CDR1, thereby increasing the hydrophilicity of the V(H):V(L) interface. To evaluate the specific contribution of Gly35 to domain properties, Gly35 was introduced into a V(H) domain with poor solution properties. This greatly enhanced the recovery of the mutant from a gel filtration matrix, but had little effect on its ability to refold reversibly after heat denaturation. Our results confirm the importance of a hydrophilic V(H):V(L) interface for purification of isolated V(H) domains, and constitute a step towards the design of isolated human V(H) domains with practical properties for immunotherapy.

Amino Acid Sequence↗
PubMed2004

Reduced global cooperativity is a common feature underlying the amyloidogenicity of pathogenic lysozyme mutations.

One of the 20 or so human amyloid diseases is associated with the deposition in vital organs of full-length mutational variants of the antibacterial protein lysozyme. Here, we report experimental data that permit a detailed comparison to be made of the behaviour of two of these amyloidogenic variants, I56T and D67H, under identical conditions. Hydrogen/deuterium exchange experiments monitored by NMR and mass spectrometry reveal that, despite their different locations and the different effects of the two mutations on the structure of the native state of lysozyme, both mutations cause a cooperative destabilisation of a remarkably similar segment of the structure, comprising in both cases the beta-domain and the adjacent C-helix. As a result, both variant proteins populate transiently a closely similar, partially unstructured intermediate in which the beta-domain and the adjacent C-helix are substantially and simultaneously unfolded, whereas the three remaining alpha-helices that form the core of the alpha-domain still have their native-like structure. We show, in addition, that the binding of a camel antibody fragment, cAb-HuL6, which was raised against wild-type lysozyme, restores to both variant proteins the stability and cooperativity characteristic of the wild-type protein; as a consequence, it inhibits the formation of amyloid fibrils by both variants. These results indicate that the reduction in global cooperativity, and the associated ability to populate transiently a specific, partly unfolded intermediate state under physiologically relevant conditions, is a common feature underlying the behaviour of these two pathogenic mutations. The formation of intermolecular interactions between lysozyme molecules that are in this partially unfolded state is therefore likely to be the fundamental trigger of the aggregation process that ultimately leads to the formation and deposition in tissue of amyloid fibrils.

Amino Acid Substitution↗