Publications

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llama single domain antibodies as a tool for molecular mimicry.in camelids, a subset of the immunoglobulins consists of heavy-chain homodimers devoid of light chains, and are thus called heavy-chain iggs (hciggs). their variable region (vhh) is the smallest antigen-binding fragment possible, and being just one polypeptide chain it is especially suitable for engineering. in particular, camelid single domain antibodies might be very useful for molecular mimicry and anti-idiotypic vaccination. in the present work, we show that llamas immunized with an anti-dna ...200515890359
prolonged in vivo residence times of llama single-domain antibody fragments in pigs by binding to porcine immunoglobulins.the therapeutic parenteral application of llama single-domain antibody fragments (vhhs) is hampered by their small size, resulting in a fast elimination from the body. here we describe a method to increase the serum half-life of vhhs in pigs by fusion to another vhh binding to porcine immunoglobulin g (pigg). we isolated 19 pigg-binding vhhs from an immunized llama using phage display. six vhhs were genetically fused to model vhh k 609 that binds to escherichia coli f4 fimbriae. all six yeast-pr ...200515992972
treatment of long bone fractures in south american camelids: 5 cases.an adult llama and four alpacas were referred with long bone fractures. the llama presented with a closed, comminuted fracture of the right metatarsal bones. two of the alpacas presented with comminuted fractures of the proximal radius. one fracture was closed and one was open. one alpaca had a closed, comminuted fracture of the distal radius, and the final alpaca had a closed, oblique fracture of the metatarsus. a diagnosis was made in each animal on the basis of clinical examination and radiog ...200516035181
fetal brain hypometabolism during prolonged hypoxaemia in the llama.in this study we looked for additional evidence to support the hypothesis that fetal llama reacts to hypoxaemia with adaptive brain hypometabolism. we determined fetal llama brain temperature, na(+) and k(+) channel density and na(+)-k(+)-atpase activity. additionally, we looked to see whether there were signs of cell death in the brain cortex of llama fetuses submitted to prolonged hypoxaemia. ten fetal llamas were instrumented under general anaesthesia to measure ph, arterial blood gases, mean ...200516037083
[mass appearance of actinomycosis in a llama herd in switzerland].this work describes the symptoms of actinomycosis (actinomyces bovis) in several llamas in a herd raised in switzerland as well as their therapy. in 5 llamas a hematological and chemical blood examination was made where low calcium, partially low iron and copper levels were found. various reasons are discussed which influence the bone metabolism and lead to a higher susceptibility of bone infection.200516128440
toxoplasma gondii infection in adult llamas (lama glama) and vicunas (vicugnavicugna) in the peruvian andean region.the present study was designed to investigate toxoplasma gondii infection in adult llamas (lama glama) and vicunas (vicugna vicugna) in the peruvian andean region, for which to date no information has been available. serum samples from 43 llamas (l. glama) and 200 vicunas were tested by ifat detecting titres of 1:50 or higher in 55.8% (33.9-70.9%) and 5.5% (2.8-9.6%), respectively. ifat titres ranged from 1:50 to 1:6400. in order to avoid cross reactions with closely related coccidian parasites ...200515893075
vasodilator tone in the llama fetus: the role of nitric oxide during normoxemia and hypoxemia.the fetal llama responds to hypoxemia, with a marked peripheral vasoconstriction but, unlike the sheep, with little or no increase in cerebral blood flow. we tested the hypothesis that the role of nitric oxide (no) may be increased during hypoxemia in this species, to counterbalance a strong vasoconstrictor effect. ten fetal llamas were operated under general anesthesia. mean arterial pressure (map), heart rate, cardiac output, total vascular resistance, blood flows, and vascular resistances in ...200515905225
in vitro and in vivo maturation of llama oocytes.cumulus-oocyte complexes (coc) were collected from abbatoir-derived llama ovaries and cultured in vitro for 28, 30, or 36 h at 39 degrees c in 5% co2 to determine the time required for maturation. the majority of coc (n=298, 87%) were classified as categories 1 and 2 (coc with > or =5 layers or 2-4 compact layers of cumulus cells, respectively) and homogeneous ooplasm, and the proportion that underwent nuclear maturation (mii) was 78, 81 and 80%, after 28, 30 and 36 h, respectively (p=0.65). to ...200515910925
assessment of serum concentrations and sedative effects of fentanyl after transdermal administration at three dosages in healthy llamas.to determine the serum concentrations and sedative effects of fentanyl after transdermal administration at 3 dosages in llamas.200515934620
llama antibodies against a lactococcal protein located at the tip of the phage tail prevent phage infection.bacteriophage p2 belongs to the most prevalent lactococcal phage group (936) responsible for considerable losses in industrial production of cheese. immunization of a llama with bacteriophage p2 led to higher titers of neutralizing heavy-chain antibodies (i.e., devoid of light chains) than of the classical type of immunoglobulins. a panel of p2-specific single-domain antibody fragments was obtained using phage display technology, from which a group of potent neutralizing antibodies were identifi ...200515968064
specific production rate of vhh antibody fragments by saccharomyces cerevisiae is correlated with growth rate, independent of nutrient limitation.saccharomyces cerevisiae carrying a multicopy integrated expression vector containing the gene encoding a llama antibody fragment, has been cultivated in continuous cultures both under carbon and nitrogen limiting conditions with galactose as the sole carbon source. vhh-r2 expression was under control of the inducible gal7 promoter. induction however, was independent of the galactose consumption rate and maximal at all growth rates. vhh-r2 was secreted with 70% efficiency at all growth rates and ...200515979755
trichophytobezoar duodenal obstruction in new world camelids.to describe clinical findings, surgical treatment, and outcome associated with trichophytobezoar duodenal obstruction in new world camelids.200516266347
production of bifunctional proteins by aspergillus awamori: llama variable heavy chain antibody fragment (v(hh)) r9 coupled to arthromyces ramosus peroxidase (arp).the arthromyces ramosus peroxidase gene (arp) was genetically fused to either the 5'- or 3'-terminal ends of the gene encoding llama variable heavy chain antibody fragment v(hh) r9, resulting in the fusion expression cassettes arp-r9 or r9-arp. aspergillus awamori transformants were obtained which produced up to 30 mgl(-1) fusion protein in the culture medium. both fusion proteins showed peroxidase activity in an abts activity test. considerable amounts of fusion protein were detected intracellu ...200516169108
escherichia coli f4 fimbriae specific llama single-domain antibody fragments effectively inhibit bacterial adhesion in vitro but poorly protect against diarrhoea.oral administration of polyclonal antibodies directed against enterotoxigenic escherichia coli (etec) f4 fimbriae is used to protect against piglet post-weaning diarrhoea. for cost reasons, we aim to replace these polyclonal antibodies by recombinant llama single-domain antibody fragments (vhhs) that can be produced efficiently in microorganisms. six f4 fimbriae specific vhhs were isolated. the vhh that was produced at the highest level by yeast, k609, was further analysed. 3.8 mg/l k609 inhibit ...200516221532
expression and production of llama variable heavy-chain antibody fragments (v(hh)s) by aspergillus awamori.we report the expression and production of llama variable heavy-chain antibody fragments (v(hh)s) by aspergillus awamori. fragments encoding v(hh)s were cloned in a suitable aspergillus expression vector and transformants secreting v(hh) fragments were analysed for integrated gene copy-numbers, mrna levels and protein production. functional v(hh)s were detected in the culture medium, indicating the feasibility of producing this type of protein in a fungal expression system. secreted v(hh)s were ...200515378291
detection of specific antibodies to neospora caninum and toxoplasma gondii in naturally infected alpacas (lama pacos), llamas (lama glama) and vicunas (lama vicugna) from peru and germany.sera of an experimentally neospora caninum infected llama and a non-infected control llama were used to establish an immunoblot, an elisa and an ifat to detect antibodies against n. caninum tachyzoites. subsequently, serum samples collected from a total of 871 south american camelids (sac: lama glama, lama pacos, lama vicugna) of two farms in peru and from 32 sac of a farm in central germany were examined for antibodies against n. caninum and toxoplasma gondii. based on the recognition of specif ...200515893073
evaluation of glucose tolerance and insulin sensitivity in ilama crias.to investigate glucose tolerance and insulin sensitivity in llama crias.200516008225
retinoblastoma in the eye of a llama (llama glama).a 6-year-old, pregnant female llama experienced a 6-month history of epiphora, buphthalmos, and acute loss of vision in the left eye. the condition was unresponsive to topical antimicrobial and anti-inflammatory therapy and progressed to corneal rupture.200516008711
a novel nanobody that detects the gain-of-function phenotype of von willebrand factor in adamts13 deficiency and von willebrand disease type 2b.von willebrand factor (vwf) is unable to interact spontaneously with platelets because this interaction requires a conversion of the vwf a1 domain into a glycoprotein ibalpha (gpibalpha) binding conformation. here, we discuss a llama-derived antibody fragment (au/vwfa-11) that specifically recognizes the gpibalpha-binding conformation. au/vwfa-11 is unable to bind vwf in solution, but efficiently interacts with ristocetin- or botrocetin-activated vwf, vwf comprising type 2b mutation r1306q, or i ...200516014562
[ethnographic study of neurological and mental diseases among the uru-chipaya peoples of the andean altiplano].the uru-chipaya people are an ethnic group of about 2,500 people, descendants of primitive andean cultures. their isolation (they live at an altitude of 4,000 metres in southern bolivia), their non-written language (chipaya-puquina) and their traditional way of life, clothing and customs, which are similar to those used for thousands of years, make this an unusual culture. the aim of our work was to carry out an ethnographic study of the neurological diseases experienced by these people, the way ...200516028191
determination of the anatomic communications among compartments within the carpus, metacarpophalangeal and metatarsophalangeal joints, stifle joint, and tarsus in llamas.to determine the anatomic communications among compartments within the carpus, metacarpophalangeal and metatarsophalangeal joints, stifle joint, and tarsus in llamas.200516173490
molecular cloning and phylogenetic analysis of inflammatory cytokines of camelidae (llama and camel).we cloned, sequenced and analyzed the cdnas encoding camelidae inflammatory cytokines, including llama (lama glama) interleukin (il)-1alpha, il-1beta, il-6, tumor necrosis factor (tnf)-alpha and camel (camelus bactrianus) il-6 and tnf-alpha. the similarity levels of the deduced amino acid sequences of il-1alpha, il-1beta, il-6 and tnf-alpha from llama (camel) to those from other mammalian species, ranged from 60.7% to 87.7%, 52.8% to 75.3%, 41.4% to 98.6%, and 72.9% to 99.6%, respectively. phylo ...200516210805
complete cdna sequences and phylogenetic analyses of the th1 and th2 cytokines of the bactrian camel (camelus bactrianus).the complementary dnas of the th1 (il-2, il-12p35, and ifn-gamma) and th2 (il-4, il-10 and il-13) cytokine genes of the bactrian camel (camelus bactrianus) were cloned, sequenced, and analyzed. il-2, il-4, il-10, il-12p35, il-13, and ifn-gamma were found to have 465, 402, 537, 669, 411, and 501 bp length open reading frames with 154, 133, 178, 222, 136, and 166 amino acid encodings, respectively. the homology ranged from 58.8% to 100% between the nucleotide sequences of the camel cytokine genes ...200617019063
malignant nonteratoid ocular medulloepithelioma in a llama (llama glama).a 6-year-old female llama presented with buphthalmos of its right eye owing to the presence of an intraocular mass. the affected globe was enucleated and submitted for microscopic examination. the intraocular mass was diagnosed as malignant medulloepithelioma. within the following months, the llama developed soft tissue masses, which completely filled the right orbital cavity and expanded the cranial portion of the right mandibular bone, and enlarged mandibular lymph nodes. euthanasia was electe ...200617037625
lactococcal bacteriophage p2 receptor-binding protein structure suggests a common ancestor gene with bacterial and mammalian viruses.lactococcus lactis is a gram-positive bacterium used extensively by the dairy industry for the manufacture of fermented milk products. the double-stranded dna bacteriophage p2 infects specific l. lactis strains using a receptor-binding protein (rbp) located at the tip of its noncontractile tail. we have solved the crystal structure of phage p2 rbp, a homotrimeric protein composed of three domains: the shoulders, a beta-sandwich attached to the phage; the neck, an interlaced beta-prism; and the r ...200616327804
selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy.we previously demonstrated that oral application of the recombinant single-domain antibody fragment (vhh) clone k609, directed against escherichia coli f4 fimbriae, reduced e. coli-induced diarrhoea in piglets, but only at high vhh doses. we have now shown that a large portion of the orally applied k609 vhh is proteolytically degraded in the stomach. stringent selection for proteolytic stability identified seven vhhs with 7- to 138-fold increased stability after in vitro incubation in gastric fl ...200616450109
therapeutic effect of llama derived vhh fragments against streptococcus mutans on the development of dental caries.streptococcus mutans is the main cause of dental caries. we evaluated the therapeutic effect of variable regions of a llama heavy chain antibody fragments directed against s. mutans named s36-vhh (s for streptococcus) alone or fused with glucose oxidase (gox) from aspergillus niger. western blot analysis and elisa revealed binding of the s36-vhh to the streptococcal antigen i/ii adhesin molecule of s. mutans serotype c. in a rat-desalivated caries model, daily administration of s36-vhh significa ...200616636830
construction and evaluation of novel fusion proteins for targeted delivery of micro particles to cellulose surfaces.the use of igg antibodies and fragments has been limited to specific sectors of the biotechnology industry due to the high cost of producing large batches of product necessary for alternative applications. a novel class of camelid antibodies, known as v(hh) offer a more economical opportunity to meet a wider application in industry. in this study, we report the evaluation of four llama v(hh)-cellulose binding domain fusion proteins displaying varying formats of v(hh) and cbd domains. proteins we ...200616673421
analytical chemistry: cause for a llama. 200616688166
unusual findings in a llama. 200617127766
identification by phage display of single-domain antibody fragments specific for the odd domain in hypoxia-inducible factor 1alpha.hypoxia triggers the transcription of genes responsible for cell survival via the key player transcription factor hypoxia-inducible factor 1alpha (hif-1alpha). overexpression of this protein has been implicated in cardiovascular disorders, carcinogenesis and cancer progression. for functional and diagnostic studies on the hif-1alpha protein, we have identified single-domain antibody fragments directed against this protein by using a llama-derived nonimmune phage display library. this library dis ...200616482104
imaging diagnosis--pulmonary metastases in new world camelids.the radiographic appearance of pulmonary metastatic disease from carcinoma is described in a llama and an alpaca. in one, a diffuse miliary pattern was seen. in the other, a more atypical unstructured interstitial pattern was recognized. metastatic pulmonary neoplasia in camelids may assume a generalized miliary or unstructured pattern.200617153066
dysautonomia and salmonellosis in an 11-year-old female llama (lama glama). 200616496946
comparison of the effect of ovulation-inducing factor (oif) in the seminal plasma of llamas, alpacas, and bulls.we have recently reported the presence of an ovulation-inducing factor (oif) in the seminal plasma of llamas and alpacas-species characterized as induced ovulators. the study was designed to test the hypothesis that the seminal plasma of bulls will induce ovulation in llamas, and to compare the ovulation-inducing effect of seminal plasma of conspecific versus hetero-specific males. the seminal plasma of alpacas, a closely related induced ovulator (lama pacos), and cattle, a distantly related rum ...200616630652
isolation and characterization of a thermally stable recombinant anti-caffeine heavy-chain antibody fragment.we have isolated and characterized a caffeine-specific, heavy-chain-only antibody fragment (v(hh)) from llama that is capable of being utilized to analyze caffeine in hot and cold beverages. camelid species (llama and camel) were selected for immunization because of their potential to make heat-stable, heavy-chain-only antibodies. llamas and camels were immunized with caffeine covalently linked to keyhole limpet hemocyanin, and recombinant antibody techniques were used to create phage displayed ...200616808459
a novel and easy method for the production of recombinant peptides for use in the generation of monospecific antisera against lama glama igg2b and igg2c subclasses.we describe an easy method for the production of small recombinant peptides of 8 amino acid residues expressed as a fusion peptide with glutathione s-transferase (gst) and employed in an immunisation schedule to obtain polyclonal antibodies. the chosen peptides corresponded to specific fragments of the hinge regions of llama (lama glama) igg2 subisotypes b (2bh) and c (2ch). the dna sequences encoding each peptide were ligated individually into pgex-5x-2, which encodes gst. once purified from a ...200616820167
design of a peptibody consisting of the antimicrobial peptide dhvar5 and a llama variable heavy-chain antibody fragment.immunoconjugates have been widely studied as potential therapeutics for infectious diseases to direct unspecific antimicrobials to pathogens. in this study, the recombinant approach was used for expression of the immunoconjugate composed of the variable domain of a llama heavy-chain antibody (vhh) against streptococcus mutans and dhvar5, a synthetic antimicrobial peptide. before cloning, the impact of the elongation of the peptide termini on its biological activity was evaluated by chemical synt ...200616882317
changes in llama (lama glama) milk composition during lactation.milk samples were collected weekly from 10 llamas during the first 27 wk after parturition under controlled stable conditions. mean values for the concentrations of the major milk components across the lactation period were 4.70% fat, 4.23% protein, 5.93% lactose, 15.61% dry matter, and 22.62 mg/dl of milk urea n. all constituents were affected by the stage of lactation. there was an increase in fat to protein ratio as protein concentration declined and fat concentration increased. fat, protein, ...200616899683
pharmacokinetics of intravenously administered caffeine in healthy alpacas (lama pacos) and llamas (lama glama).to determine the pharmacokinetic disposition of iv administered caffeine in healthy lama spp camelids.200616740103
llama single-chain antibody that blocks lipopolysaccharide binding and signaling: prospects for therapeutic applications.sepsis is a considerable health problem and a burden on the health care system. endotoxin, or lipopolysaccharide (lps), present in the outer membrane of gram-negative bacteria, is responsible for more than 50% of the sepsis cases and is, therefore, a legitimate target for therapeutic approaches against sepsis. in this study, we selected and characterized a llama single-chain antibody fragment (vhh) directed to neisseria meningitidis lps. the vhh, designated vhh 5g, showed affinity to purified lp ...200616928888
clinical, humoral, and pathologic findings in adult alpacas with experimentally induced corynebacterium pseudotuberculosis infection.to experimentally infect adult alpacas by id inoculation of corynebacterium pseudotuberculosis, follow the clinical and pathologic course of disease, and study the humoral response to infection.200616948603
improving solubility and refolding efficiency of human v(h)s by a novel mutational approach.the antibody v(h) domains of camelids tend to be soluble and to resist aggregation, in contrast to human v(h) domains. for immunotherapy, attempts have therefore been made to improve the properties of human v(h)s by camelization of a small set of framework residues. here, we have identified through sequence comparison of well-folded llama v(h) domains an alternative set of residues (not typically camelid) for mutation. thus, the solubility and thermal refolding efficiency of a typical human v(h) ...200616971398
receptor-binding protein of lactococcus lactis phages: identification and characterization of the saccharide receptor-binding site.phage p2, a member of the lactococcal 936 phage species, infects lactococcus lactis strains by binding initially to specific carbohydrate receptors using its receptor-binding protein (rbp). the structures of p2 rbp, a homotrimeric protein composed of three domains, and of its complex with a neutralizing llama vh domain (vhh5) have been determined (s. spinelli, a. desmyter, c. t. verrips, h. j. de haard, s. moineau, and c. cambillau, nat. struct. mol. biol. 13:85-89, 2006). here, we show that vhh ...200616547026
compressive myelopathy due to intervertebral disk extrusion in a llama (lama glama).a 12-year-old intact female llama was euthanized following acute onset of spastic tetraparesis and recumbency with inability to rise. postmortem examination revealed caudal cervical spinal cord compression due to a mass within the ventral spinal canal arising from the c6-c7 intervertebral disk space and attached to an irregularly thickened annulus fibrosis. on histopathologic examination, the mass was composed of amorphous acellular basophilic to amphophilic material admixed with irregularly arr ...200616566272
reduction in morbidity of rotavirus induced diarrhoea in mice by yeast produced monovalent llama-derived antibody fragments.apart from the use of oral rehydration solution, there are currently no treatment modalities for rotavirus induced diarrhoea, which is particularly relevant to developing countries. fragments derived from llama heavy chain antibodies were previously shown to be highly stable, efficiently produced in yeast and exhibiting high epitope specific affinity. we now aim to demonstrate that these antibody fragments are capable of reducing morbidity of rotavirus induced diarrhoea. here we show the isolati ...200616616802
pulmonary hypertension and right-sided heart failure in an adult llama with hepatic disease.a 13-year-old llama was examined because of lethargy, inappetence, and syncope.200616506943
postal survey of the population of south american camelids in the united kingdom in 2000/01.the members of the two leading british camelid breeders associations were surveyed by means of a postal questionnaire between december 2000 and january 2001; 696 questionnaires were posted and 218 usable responses were returned. a total of 3520 camelids were recorded, of which 2719 (77.2 per cent) were alpacas (lama pacos) and 726 (20.6 per cent) were llamas (lama glama). ninety-four per cent of the camelid herds were of one species, and 70 per cent of the animals were kept for more than one pur ...200616428662
facile generation of heat-stable antiviral and antitoxin single domain antibodies from a semisynthetic llama library.llamas possess a class of unconventional immunoglobulins that have only heavy chains; unpaired heavy variable domains are responsible for antigen binding. these domains have previously been cloned and expressed as single domain antibodies (sdabs); they comprise the smallest known antigen binding fragments. sdabs have been shown to bind antigens at >90 degrees c and to refold after being denatured. to take advantage of the remarkable properties of sdabs, we constructed a large, semisynthetic llam ...200617165813
ablynx makes nanobodies from llama bodies. 200617185218
generation of heavy-chain-only antibodies in mice.we have generated transgenic mice containing hybrid llama/human antibody loci that contain two llama variable regions and the human d, j, and cmu and/or cgamma constant regions. such loci rearrange productively and rescue b cell development efficiently without lc rearrangement. heavy-chain-only antibodies (hcab) are expressed at high levels, provided that the ch1 domain is deleted from the constant regions. hcab production does not require an igm stage for effective pre-b cell signaling. antigen ...200617015837
identification of single-domain, bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis.bax is a proapoptotic protein implicated in cell death involved in several neurodegenerative diseases. intracellularly expressed antibody (ab) fragments (intrabodies) inhibiting bax function would have potential for developing therapeutics for the aforementioned diseases and can serve as research tools. we report identification, cloning, and functional characterization of several bax-specific single-domain antibodies (sdabs). these minimal size ab fragments, which were isolated from a llama v(h) ...200617060401
lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea.rotavirus-induced diarrhea poses a worldwide medical problem in causing substantial morbidity and mortality among children in developing countries. we therefore developed a system for passive immunotherapy in which recombinant lactobacilli constitutively express neutralizing variable domain of llama heavy-chain (vhh) antibody fragments against rotavirus.200617083044
selection, characterization, and cdr shuffling of naive llama single-domain antibodies selected against auxin and their cross-reactivity with auxinic herbicides from four chemical families.indoleacetic acid (iaa)-binding single-domain antibodies (sdabs) were isolated from a naive phage-display library constructed from the heavy chain antibody repertoire of a ilama. the highest-affinity sdab isolated (csf2a) had a k(d) of 5-20 microm for two iaa-protein conjugates and a k(d) of 20 microm for free iaa. this sdab also bound to a synthetic auxin analogue, 1-naphthaleneacetic acid (naa), and to six auxinic herbicides (k(d) values of 0.5-2 mm), but not to serotonin and tryptophan, which ...200619127743
a novel pentamer versus pentamer approach to generating neutralizers of verotoxin 1.verotoxins (vts), or shiga-like toxins, are produced by enterohemorrhagic escherichia coli (ehec), which cause hemorrhagic colitis and hemolytic uremic syndrome. vts are the major virulence factors in ehec infection due to their cytotoxicity to various types of cells. here, we present a novel type of vt neutralizer based on pentavalent single-domain antibodies, or pentabodies. two single-domain antibodies (sdabs) specific for the receptor binding sites of the b subunit of vt1 (vt1b) were isolate ...200717134756
meat fatty acid composition of llama (lama glama) reared in the andean highlands.this study reports the results of the chemical analysis of the longissimus thoracis and lumborum taken from 20 llama males, reared in the andean highlands. the animals were slaughtered at 25 months and had a mean final body weight of 63kg. llama meat shows a low fat (3.51%) and cholesterol content (56.29mg/100g). the fatty acid composition in llama meat contains 50.34% saturated fatty acids, 42.48% monounsaturated fatty acids and 7.18% polyunsaturated fatty acids. llama meat appears to be a heal ...200722063670
tenderness evaluation and mineral levels of llama (lama glama) and alpaca (lama pacos) meat.tenderness and mineral levels were determined in the longissimus thoracis taken from 20 llama and 30 alpaca males reared in peru and slaughtered at 25 months of age. mineral contents were determined using an inductively coupled plasma emission spectrometer. tenderness evaluation was determined two and seven days post slaughter using a warner-bratzler shear force device. potassium is the mineral with the highest content, with a significant difference (p<0.05) between the two species of camelids. ...200722061947
estradiol production by preimplantation blastocysts and increased serum progesterone following estradiol treatment in llamas.estradiol is a potential candidate for the blastocyst signal responsible for maternal recognition of pregnancy in the llama (lama glama). two experiments were conducted to determine if the llama blastocyst produces estradiol during the presumed period of maternal recognition of pregnancy and if exogenous estradiol can extend the luteal phase. in experiment 1, llamas were superovulated with ecg and mated 7 days later (day 0=day of mating). blastocysts were collected nonsurgically on days 7, 9, or ...200717116376
prevalence of neoplasia in llamas and alpacas (oregon state university, 2001-2006).prevalence and type of neoplastic disease were determined in 551 camelid submissions (368 alpacas [lama pacos], 180 llamas [lama glama], and 3 cases in which species was not identified) over a 5-year period. forty neoplasms were identified in 38 animals (6.9%). prevalence of neoplasia in llamas was higher (11%) than in alpacas (4.9%). mean age of camelids with neoplasia was 9.42 +/- 4.9 years. mean age of alpacas with neoplasia (5.48 +/- 3.7 years) was significantly less than of llamas with neop ...200717402619
immunobiological role of llama heavy-chain antibodies against a bacterial beta-lactamase.in 1993, a fraction of antibodies (abs) devoid of l chain was found naturally occurring in the camelidae. they were found to lack l chains, as well as the first constant heavy-chain domain (ch(1)) and therefore they were named "heavy-chain abs" (hcabs). subsequent studies focused on the functional, structural and biochemical properties of recombinant variable fragments (rvhhs) of hcabs. it was stated that rvhhs have an augmented capacity to interact with "partially hidden" epitopes, like enzymes ...200717448545
a fungal granuloma of the frontal sinus in a llama.a 12-year-old, castrated male llama (lama glama) presented with a 12-cm diameter cranial mass. computed tomography and postmortem examination revealed that the mass invaded the calvarium and compressed the rostral part of the brain. light microscopic examination confirmed a fungal granuloma.200717966336
human and animal infections with mycobacterium microti, scotland.during 1994-2005, we isolated mycobacterium microti from 5 animals and 4 humans. only 1 person was immunocompromised. spoligotyping showed 3 patterns: vole type, llama type, and a new variant llama type.200718258049
measurement of milk intake in suckling llamas (lama glama) using deuterium oxide dilution.the objective of the study was to estimate daily milk intake in llama crias and relate nutrient intakes at peak lactation to growth data. milk intake in 11 suckling llamas was estimated from water kinetics using deuterium oxide (d2o) at d 17, 66, and 128 postpartum. daily milk intakes averaged 2.6, 2.3, and 2.0 kg at 17, 66, and 128 d postpartum, respectively. milk intake decreased with age when expressed as daily amount, percentage of body weight (bw), or per kilogram of metabolic size, but the ...200717235163
detection of anti-brucella antibodies in llama (llama glama).seven llamas were immunized with killed brucella abortus s1119.3 cells and bled sequentially, resulting in 64 samples. an eighth llama was kept as a negative control. in addition, 299 llama and 2075 alpaca sera, submitted for diagnostic testing, were included. sera from all llamas were tested by the buffered antigen plate agglutination test, the complement fixation test, and the indirect enzyme immunoassays using smooth and rough lipopolysaccharides. a competitive enzyme immunoassay and fluoresc ...200717236397
isolation, characterization and pentamerization of alpha-cobrotoxin specific single-domain antibodies from a naïve phage display library: preliminary findings for antivenom development.conventional antivenoms to snakebite generated from the serum of immunized animals, often elicit adverse reactions and have mismatched pharmacokinetic profiles with their target toxins due to antibody/toxin size discrepancies which results in poor neutralization. furthermore, animal immunization protocols are often lengthy and have batch to batch variability. recombinant v(h)h-based antivenoms may help overcome these problems. three v(h)h fragments with specificity to alpha-cobrotoxin, a snake n ...200717257638
changes in testicular histology and sperm quality in llamas (lama glama), following exposure to high ambient temperature.the aim of the study was to investigate whether a moderately elevated ambient temperature (29 degrees c, 4 weeks, 24hperday) has an effect on the spermatogenesis in male llamas (lama glama) and to monitor the recrudescence of spermatogenesis. thirteen llamas were used. semen parameters were monitored in four of the llamas and six animals were castrated at different times after the heat treatment. an additional three llamas were used as control animals and were castrated without any treatment. sp ...200717368737
distribution of the internal pudendal artery in male and female llama (lama glama).the aim of this research has been to describe the internal pudendal artery distribution in male and female llama and to compare it with that of other domestic animals including the one-humped camel (camelus dromedarius). the arterial system was perfused with a solution of 14% coloured plaster and preserved in a solution of a 10% formalin, 3% carbolic acid and 3% glycerine. the systematic dissection was made using traditional working techniques and standard instruments. the internal pudendal arte ...200717534039
single domain antibodies from llama effectively and specifically block t cell ecto-adp-ribosyltransferase art2.2 in vivo.the purpose of our study was to develop a tool for blocking the function of a specific leukocyte ecto-enzyme in vivo. art2.2 is a toxin-related ecto-enzyme that transfers the adp-ribose moiety from nad onto other cell surface proteins. art2.2 induces t cell death by activating the cytolytic p2x7 purinoceptor via adp-ribosylation. here, we report the generation of art2.2-blocking single domain antibodies from an immunized llama. the variable domain of heavy-chain antibodies (vhh domain) represent ...200717575259
evolving in thin air--lessons from the llama fetus in the altiplano.compared with lowland species, fetal life for mammalian species whose mothers live in high altitude is demanding. for instance, fetal llamas have to cope with the low fetal arterial po2 of all species, but also the likely superimposition of hypoxia as a result of the decreased oxygen environment in which the mother lives in the andean altiplano. when subjected to acute hypoxia the llama fetus responds with an intense peripheral vasoconstriction mediated by alpha-adrenergic mechanisms plus high p ...200717588504
carcinoma in a mixed mammary tumor in a llama (lama glama).a 13-year-old female llama was presented to the referring veterinarian for swelling and firmness of the right rear mammary gland, for a duration of 2 months, which had been unresponsive to antibiotics. a formalin-fixed wedge biopsy specimen from the affected quarter was submitted to purdue university animal disease diagnostic laboratory for histopathology. histopathologic examination revealed tubulopapillary acinar or solid nest-like clusters of neoplastic epithelial cells surrounded by whorls a ...200717609363
antibody responses in new world camelids with tuberculosis caused by mycobacterium microti.antibody responses in new world camelids (nwc) infected with mycobacterium microti were studied by two serological methods, multiantigen print immunoassay (mapia) and lateral-flow-based rapid test (rt). serum samples were collected during 2004-2006 from 87 animals including 1 alpaca and 7 llamas with confirmed or suspected m. microti infection, 33 potentially exposed but clinically healthy animals from known infected herds, and 46 control nwc from herds where infection had not been previously di ...200717628360
use of flow cytometry to identify monoclonal antibodies that recognize conserved epitopes on orthologous leukocyte differentiation antigens in goats, llamas, and rabbits.flow cytometry was used to screen a panel of 320 mabs, submitted to the animal homologues section of the hlda8, for mabs that recognize epitopes conserved on orthologous leukocyte differentiation antigens (lda) in goats, lamas, and rabbits. nineteen mabs specific for cd11a (1), cd14 (3), cd18 (1), cd21 (1), cd29 (2), cd44 (2), cd47 (3), cd49d (1), cd172a (1), cd45rb (1), cd61 (1), ract48a, and gbsp71a reacted with goat lda. twenty three mabs specific for cd7 (1), cd9 (2), cd11a (1), cd14 (3), cd ...200717686528
lactobacillli expressing llama vhh fragments neutralise lactococcus phages.bacteriophages infecting lactic acid bacteria (lab) are widely acknowledged as the main cause of milk fermentation failures. in this study, we describe the surface-expression as well as the secretion of two functional llama heavy-chain antibody fragments, one binding to the major capsid protein (mcp) and the other to the receptor-binding proteins (rbp) of the lactococcal bacteriophage p2, by lactobacilli in order to neutralise lactococcal phages.200717875214
selection of genetically encoded fluorescent single domain antibodies engineered for efficient expression in escherichia coli.genetically encoded fluorescent antibodies are desirable for many applications in biotechnology and proteomics. however, the efficient production of single chain antibodies fused to fluorescent proteins like green fluorescent protein is still a major challenge. this is due to the opposite redox requirements of recombinant antibodies and fluorescent proteins for proper folding. to overcome this obstacle, we designed a novel strategy to directly select functional fluorescent antibodies (fluobodies ...200717921141
stabilization of an immunoglobulin fold domain by an engineered disulfide bond at the buried hydrophobic region.we report for the first time the stabilization of an immunoglobulin fold domain by an engineered disulfide bond. in the llama single-domain antibody, which has human chorionic gonadotropin as its specific antigen, ala49 and ile70 are buried in the structure. a mutant with an artificial disulfide bond at this position showed a 10 degrees c higher midpoint temperature of thermal unfolding than that without the extra disulfide bond. the modified domains exhibited an antigen binding affinity compara ...200717932041
adaptations of subpalpebral lavage systems used for llamas (lama glama) and a harbor seal (phoca vitulina).subpalpebral lavage systems (splss) were adapted for use in zoo llamas (lama glama) and a wild harbor seal (phoca vitulina) during therapy for severe ulcerative keratitis or corneal perforation. one llama presented with a melting corneal ulcer caused by pseudomonas aeruginosa, which necessitated frequent application of a topical ophthalmic antibiotic. the lavage system was used routinely during the day and was connected to a balloon infusion system at night to allow for continuous medication adm ...200717939355
rapid assembly of sensitive antigen-capture assays for marburg virus, using in vitro selection of llama single-domain antibodies, at biosafety level 4.there is a pressing need for rapid and reliable approaches to the delivery of sensitive yet rugged diagnostic assays specific for emerging viruses, to hasten containment of outbreaks when and wherever they occur. within 3 weeks, we delivered an antigen-capture assay for marburg virus (marv) that was based on llama single-domain antibodies (sdabs) selected at biosafety level 4. four unique sdabs were capable of independently detecting marv variants musoke, ravn, and angola without cross-reactivit ...200717940952
[congenital malformation of the front limbs in a llama (lama glama): polydactyly in combination with arthrogryposis].a male, 10 month old llama with malformation of both front limbs was presented. both front limbs had one more digit located medially. the distal phalanx of this additional digit at the left front limb reached the ground by the tip of the keratinzed pad and the toenail. the accessory digit at the right front limb was bent in a 90 degree angle caudolateraly. beside the digital bones of the accessory digits the second metacarpal bone and the first carpal bone could be detected in both front limbs b ...200718085162
multiplexed fluid array screening of phage displayed anti-ricin single domain antibodies for rapid assessment of specificity.phage display is a well-known technique that facilitates the selection of peptides or proteins that bind to a desired target. using this tool, binding elements contained in the natural immune repertoires of the source animal or from a synthetically generated collection may be selected. the unpaired variable domain of the llama's heavy-chain-only classes of immunoglobulins represents an ideal source of genetic material to create phage display libraries. initial panning of a semi-synthetic llama l ...200718251258
effect of ovarian superstimulation on coc collection and maturation in alpacas.the objective of the present study was to compare the ovarian follicular response, cumulus-oocyte complex (coc) collection rate, and maturational status of coc collected from alpacas subsequent to treatment with two different superstimulatory protocols. alpacas (n=7 per group) were treated with: (1) 200mg of fsh im divided bid for 3d, plus a single i.v. dose of 1000iu hcg 24h after the last fsh treatment, or (2) 1200iu of ecg as a single i.m. dose, plus a single i.v. dose of 1000iu of hcg on day ...200716530992
measurements of the bodyweight and other physical characteristics of 11 llamas (lama glama) from birth to weaning.the bodyweight (bw), thoracic circumference, length and height of 11 llamas were recorded weekly from birth for 27 weeks. their mean (sd) birth weight was 11.60 (2.35) kg and their weight increased in curvilinear fashion reaching 61.05 (13.75) kg after 27 weeks. their average daily gains reached the highest value three weeks after birth and then decreased, reaching the lowest value of 262 g per day at 27 weeks. on a metabolic bw(0.83) basis, the growth of the llamas when their dams were at peak ...200717938410
generation of llama single-domain antibodies against methotrexate, a prototypical hapten.single-domain antibodies specific to methotrexate (mtx) were obtained after immunization of one llama (llama glama). specific vhh domains (v-d-j-region) were selected by panning from an immune-llama library using phage display technology. the antibody fragments specific to mtx were purified from escherichia coli (c41 strain) periplasm by immobilized metal affinity chromatography with an expression level of around 10mg/l. a single band around 16,000da corresponding to vhh fragments was found afte ...200717011035
evaluation of neospora caninum and toxoplasma gondii infections in alpaca (vicugna pacos) and llama (lama glama) aborted foetuses from peru.the aim of this study was to investigate the participation of neospora caninum and toxoplasma gondii in abortion cases of peruvian llamas and alpacas. fifteen aborted foetuses were recovered from two main rearing areas of camelids in peru (central or south andean region). foetal histopathology was used to detect the presence of protozoal-associated lesions in target organs. n. caninum and t. gondii infections were confirmed by immunohistochemistry (ihc) combined with pcr and by pcr alone, respec ...200717931783
eastern equine encephalitis in 9 south american camelids.eastern equine encephalitis (eee) virus is a mosquito-borne togavirus (alphavirus) that causes severe (often fatal) encephalitis in many mammalian species, but it has not been reported previously in south american camelids. hypothesis: south american camelids can become naturally infected with eee virus and show encephalitic signs similar to those observed in other affected species.200717708408
in vitro production of llama (lama glama) embryos by intracytoplasmic sperm injection: effect of chemical activation treatments and culture conditions.assisted reproductive technologies in the llama (lama glama) are needed to provide alternative methods for the propagation, selection and genetic improvement; however, recovery of adequate quantity and quality of spermatozoa for conventional ivf is problematic. therefore, an effort was made to adapt the intracytoplasmic sperm injection (icsi) procedure for the in vitro production of llama embryos. the specific objectives of this study were: (1) to determine in vitro maturation rates of oocytes r ...200716846701
morphologic and biochemical characterization of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas.to determine the morphologic and biochemical characteristics of hyperextension of the metacarpophalangeal and metatarsophalangeal joints in llamas.200717669028
use of grid keratotomy for the treatment of indolent corneal ulcer in a llama.a case of indolent corneal ulcer in a llama (llama glama) is described. diagnostic testing included a complete ophthalmic examination with corneal cytologic and histopathologic examination. successful management involved grid keratotomy and topical application of neomycin-polymixin-bacitracin and atropine 1% ointments. weekly follow-up examinations are described until healing was considered complete.200717494370
secondary hepatogenous photosensitization in a llama (lama glama) bred in the state of sáo paulo, brazil. 200717436914
humerus fractures in llamas and alpacas: seven cases (1998-2004).to describe treatment and outcome of humerus fractures in llamas and alpacas.200717214823
surgical repair of femoral fractures in new world camelids: five cases (1996-2003).five new world camelids were admitted to the western college of veterinary medicine between 1996 and 2003 for evaluation of femoral fractures. there were three alpacas and two llamas. four of the animals were female and three were less than 3 months of age. fracture configurations consisted of distal physeal fractures (three), a comminuted diaphyseal/metaphyseal fracture, and a transverse diaphyseal fracture. fractures were diagnosed with a combination of physical examination and radiographs in ...200717397385
passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease.foot-and-mouth disease (fmd) is a highly contagious disease that occasionally causes outbreaks in europe. there is a need for therapies that provide rapid protection against fmd in outbreak situations. we aim to provide such rapid protection by passive immunization with llama single-domain antibody fragments (vhhs). twenty-four vhhs binding serotype o fmdv in vitro were isolated from immunized llamas by phage display and expressed in bakers yeast for further characterization. they recognized fou ...200717127019
anaerobic co-digestion of aquatic flora and quinoa with manures from bolivian altiplano.quinoa stalk (chenopodium quinoa willd.) from agricultural crop residue, totora (schoenoplectus tatora) and o-macrophytes (aquatic flora) from lake titicaca (on the bolivian altiplano) were studied in a wet anaerobic co-digestion process together with manure from llama, cow and sheep. anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 degrees c with 30 days of hydraulic retention time (hrt) and an organic loading rate (olr) of 1.8 kg vs m(-3) d(-1). ...200818155895
conditional inactivation of hif-1 using intrabodies.hypoxia is a hallmark of solid cancers and triggers the transcription of genes responsible for cell survival. the transcription factor hypoxia-inducible factor 1 (hif-1) is a key regulator in this response and frequently activated in human cancer. hif-1 activation is associated with tumor aggressiveness and poor clinical outcome and, therefore, may provide an attractive therapeutic target. here we provide a novel approach for hif-1 targeted therapy using single-domain llama antibodies directed a ...200818791271
design of a village breeding programme for a llama population in the high andes of bolivia.no national breeding programme for llamas is in place in bolivia. initiatives for genetic improvement are rarely found and are usually carried out by ngos working in rural development or improvement of livestock production or research stations. farmers in the province of ayopaya in the district of cochabamba have formed a breeders' organization with the aim of improving fibre production. in this study, a detailed outline of a breeding programme with a focus on organizational and technical detail ...200818803786
llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (hiv-1)-neutralizing properties and high affinity for hiv-1 gp120.members of the camelidae family produce immunoglobulins devoid of light chains. we have characterized variable domains of these heavy chain antibodies, the vhh, from llamas immunized with human immunodeficiency virus type 1 (hiv-1) envelope protein gp120 in order to identify vhh that can inhibit hiv-1 infection. to increase the chances of isolating neutralizing vhh, we employed a functional selection approach, involving panning of phage libraries expressing the vhh repertoire on recombinant gp12 ...200818842738
simple sequence repeats for genetic studies of alpaca.trace sequences from the 2x alpaca genome sequencing effort were examined to identify simple sequence repeats (microsatellites) for genetic studies. a total of 6,685 repeat-containing sequences were downloaded from genbank, processed, and assembled into contigs representing an estimated 4,278 distinct sequences. this sequence set contained 2,290 sequences of length > 100 nucleotides that contained microsatellites of length > or = 14 dinucleotide or 10 trinucleotide repeats with purity equal to 1 ...200818855250
thermostable llama single domain antibodies for detection of botulinum a neurotoxin complex.immunoglobulins from animals of the camelidae family boast unique forms that do not incorporate light chains. antigen binding in these unconventional heavy-chain homodimers is mediated through a single variable domain. when expressed recombinantly these variable domains are termed single domain antibodies (sdab) and are among the smallest naturally igg-derived antigen binding units. sdab possess good solubility, thermostability, and can refold after heat and chemical denaturation making them pro ...200818947189
relevance of the diversity among members of the trypanosoma cruzi trans-sialidase family analyzed with camelids single-domain antibodies.the sialic acid present in the protective surface mucin coat of trypanosoma cruzi is added by a membrane anchored trans-sialidase (tcts), a modified sialidase that is expressed from a large gene family. in this work, we analyzed single domain camelid antibodies produced against trans-sialidase. llamas were immunized with a recombinant trans-sialidase and inhibitory single-domain antibody fragments were obtained by phage display selection, taking advantage of a screening strategy using an inhibit ...200818949046
placental glycosylation in a cama (camel-llama cross) and its relevance to successful hybridisation. 200818950721
parasite-induced leucomyelopathy in llamas (lama glama). 200818953077
probing of actinobacillus pleuropneumoniae apxiiia toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells.abstract:200819046441
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