Heat shock proteins are ubiquitous, induced under a number of environmental and metabolic stresses, with highly conserved DNA sequences among mammalian species. Camelus dromedaries (the Arabian camel) domesticated under semi-desert environments, is well adapted to tolerate and survive against severe drought and high temperatures for extended periods. This is the first report of molecular cloning and characterization of full length cDNA of encoding a putative stress-induced heat shock HSPA6 protein (also called HSP70B′) from Arabian camel. A full-length cDNA (2417 bp) was obtained by rapid amplification of cDNA ends (RACE) and cloned in pET-b expression vector. The sequence analysis of HSPA6 gene showed 1932 bp-long open reading frame encoding 643 amino acids. The complete cDNA sequence of the Arabian camel HSPA6 gene was submitted to NCBI GeneBank (accession number HQ214118.1). The BLAST analysis indicated that C. dromedaries HSPA6 gene nucleotides shared high similarity (77–91%) with heat shock gene nucleotide of other mammals. The deduced 643 amino acid sequences (accession number ADO12067.1) showed that the predicted protein has an estimated molecular weight of 70.5 kDa with a predicted isoelectric point (pI) of 6.0. The comparative analyses of camel HSPA6 protein sequences with other mammalian heat shock proteins (HSPs) showed high identity (80–94%). Predicted camel HSPA6 protein structure using Protein 3D structural analysis high similarities with human and mouse HSPs. Taken together, this study indicates that the cDNA sequences of HSPA6 gene and its amino acid and protein structure from the Arabian camel are highly conserved and have similarities with other mammalian species.
Arabian camel; molecular cloning; HSPA6; sequence characterization; cDNA cloning; 3D structure; alignment; RACE; real-time PCR
This study evaluated the use of ultrasonography for the diagnosis of Johne’s disease in camels (Camelus dromedarius). Seventy camels with confirmed Johne’s disease were examined by ultrasonography and subsequent necropsy; 15 healthy camels were included as controls. The most outstanding findings were visible enlargement of the mesenteric lymph nodes in 52 (74%) camels. Lesions had either echogenic (26%; n = 18) or anechoic (69%; n = 48) capsule and the contents were either anechoic (21%; n = 15), echogenic (27%; n = 19), or heterogeneous (46%; n = 32). Clumps of echogenic tissue interspersed with fluid pockets were imaged between the intestinal loops in 9 (13%) camels. There was mild, moderate, or severe thickening and corrugation of the intestinal wall, excessive anechoic fluid in the abdominal cavity in 18 (26%) camels, increased hepatic brightness in 30 (43%) camels, and pericardial and pleural effusions in 22 (31%) camels. Sensitivity values for detecting intestinal lesions and enlarged mesenteric lymph nodes were 95% and 84%, respectively.
The evolutionary relationship between the domestic bactrian camel and the extant wild two-humped camel and the factual origin of the domestic bactrian camel remain elusive. We determined the sequence of mitochondrial cytb gene from 21 camel samples, including 18 domestic camels (three Camelus bactrianus xinjiang, three Camelus bactrianus sunite, three Camelus bactrianus alashan, three Camelus bactrianus red, three Camelus bactrianus brown and three Camelus bactrianus normal) and three wild camels (Camelus bactrianus ferus). Our phylogenetic analyses revealed that the extant wild two-humped camel may not share a common ancestor with the domestic bactrian camel and they are not the same subspecies at least in their maternal origins. Molecular clock analysis based on complete mitochondrial genome sequences indicated that the sub-speciation of the two lineages had begun in the early Pleistocene, about 0.7 million years ago. According to the archaeological dating of the earliest known two-humped camel domestication (5000–6000 years ago), we could conclude that the extant wild camel is a separate lineage but not the direct progenitor of the domestic bactrian camel. Further phylogenetic analysis suggested that the bactrian camel appeared monophyletic in evolutionary origin and that the domestic bactrian camel could originate from a single wild population. The data presented here show how conservation strategies should be implemented to protect the critically endangered wild camel, as it is the last extant form of the wild tribe Camelina.
bactrian camel; domestication; mitochondrial genome; phylogeny
To describe a novel and simple technique (STAR: Simple Targeted Arterial Rendering) to visualize the fetal cardiac outflow tracts from dataset volumes obtained with spatiotemporal image correlation (STIC) and applying a new display technology (OmniView).
We developed a technique to image the outflow tracts by drawing three dissecting lines through the four-chamber view of the heart contained in a STIC volume dataset. Each line generated the following plane: 1) Line 1: ventricular septum “en face” with both great vessels (pulmonary artery anterior to the aorta); 2) Line 2: pulmonary artery with continuation into the longitudinal view of the ductal arch; and 3) Line 3: long axis view of the aorta arising from the left ventricle. The pattern formed by all 3 lines intersecting approximately through the crux of the heart resembles a “star”. The technique was then tested in 50 normal hearts (15.3 – 40.4 weeks of gestation). To determine if the technique could identify planes that departed from the normal images, we tested the technique in 4 cases with proven congenital heart defects (ventricular septal defect, transposition of great vessels, tetralogy of Fallot, and pulmonary atresia with intact ventricular septum).
The STAR technique was able to generate the intended planes in all 50 normal cases. In the abnormal cases, the STAR technique allowed identification of the ventricular septal defect, demonstrated great vessel anomalies, and displayed views that deviated from what was expected from the examination of normal hearts.
This novel and simple technique can be used to visualize the outflow tracts and ventricular septum “en face” in normal fetal hearts. The inability to obtain expected views or the appearance of abnormal views in the generated planes should raise the index of suspicion for congenital heart disease involving the great vessels and/or the ventricular septum. The STAR technique may simplify examination of the fetal heart and could reduce operator dependency.
STIC; aorta; pulmonary artery; prenatal diagnosis; ventricular septum; fetal echocardiography; congenital heart disease; ultrasound
Haemoglobin from Camelus dromedarius provides an interesting case study of adaptation to life in deserts at extremely high temperatures. An ambition to unravel the integrated structural and functional aspects of the casual survival of this animal at high temperatures led the authors to specifically work on this problem. This work reports the preliminary crystallographic study of camel haemoglobin.
Haemoglobin is a prototypical allosteric protein that is mainly involved in the transportation of oxygen from the lungs to tissues and of carbon dioxide back to the lungs in an intrinsically coordinated manner to maintain the viability of cells. Haemoglobin from Camelus dromedarius provides an interesting case study of adaptation to life in deserts at extremely high temperatures. An ambition to unravel the integrated structural and functional aspects of the casual survival of this animal at high temperatures led us to specifically work on this problem. The present work reports the preliminary crystallographic study of camel haemoglobin. Camel blood was collected and the haemoglobin was purified by anion-exchange chromatography and crystallized using the hanging-drop vapour-diffusion method under buffered high salt concentration using PEG 3350 as a precipitant. Intensity data were collected using a MAR 345 dtb image-plate detector system. Camel haemoglobin crystallized in the monoclinic space group P21, with one whole biological molecule (α2β2) in the asymmetric unit and unit-cell parameters a = 52.759, b = 116.782, c = 52.807 Å, β = 120.07°.
haemoglobin; Camelus dromedarius; oxygen affinity
Left atrial dimensions were measured using cross sectional echocardiography in 37 patients with mitral valve disease and 30 normal subjects of similar ages. The anteroposterior (AP), superior-inferior (SI), and medial-lateral (ML) left atrial dimensions were determined at the end of ventricular systole using parasternal long and short axis and apical four chamber views (for SIa and MLa). To assess the reliability of these measurements cross sectional echocardiographic and angiographic left atrial volumes were compared in 19 patients with mitral valve disease, giving an excellent correlation. A moderate correlation was found between the anteroposterior dimension of the left atrium obtained using M mode echocardiography and that obtained using the parasternal short axis and long axis projections. In normal subjects a good correlation was found between SI and ML dimensions, while a lower correlation was found between SI and AP, and ML and AP dimensions. The SI dimension was the major axis of the left atrium and AP dimension the minor axis. In patients with mitral valve disease a good correlation was found between SI and ML dimensions, while SI and ML dimensions had a low correlation with AP dimensions. The AP dimension was the minor axis of the left atrium, while the SI and ML dimensions were not significantly different. All left atrial dimensions were significantly greater in patients with mitral valve disease than in normal subjects. Of 30 patients with at least one dimension increased, all three dimensions were abnormal in 16, two dimensions were increased in 10, and only one dimension was increased in four. AP, SI, and ML dimensions were abnormal in 25, 20, and 27 patients, respectively. Cross sectional echocardiography may provide a reliable estimate of left atrial dimensions. In patients with mitral valve disease a thorough examination of the left atrium using multiple cross sectional views is necessary to detect asymmetric left atrial enlargement and to measure the degree of left atrial dilatation.
The prevalence of Sarcocystis spp. was investigated by gross and histopathological examinations in 250 camels (Camelus dromedarius) slaughtered from 2002 to 2005 in the Mashhad Slaughterhouse, eastern Iran. Samples were taken from the diaphragm, heart, tongue, esophagus and masseter muscles for histopathological studies. No macroscopic sarcocysts were found in the samples at gross inspection. Sarcocysts were detected in 209 of 250 (83.6%) examined camels at histopathological level. The infection rate of the esophagus, heart, masseter muscles, diaphragm, and tongue was 58.8%, 48.0%, 46.8%, 41.6%, and 28.0%, respectively. There was no significant difference in the rate of infection between male (85.8%) and female (81.0%) camels. The tissue response to vital cysts was minimal; however, reaction to the degenerating cysts was severe and caused tissue damages resulting in hyperemia, hemorrhages, mononuclear cell infiltration, necrotic changes, and fibrosis. The wild and domestic carnivores especially dogs may be the final hosts of Sarcocystis spp. in this area.
Sarcocystis; camel; Iran; pathology; prevalence
To describe a novel and simple algorithm (FAST Echo: Four chamber view And Swing Technique) to visualize standard diagnostic planes of fetal echocardiography from dataset volumes obtained with spatiotemporal image correlation (STIC) and applying a new display technology (OmniView).
We developed an algorithm to image standard fetal echocardiographic planes by drawing four dissecting lines through the longitudinal view of the ductal arch contained in a STIC volume dataset. Three of the lines are locked to provide simultaneous visualization of targeted planes, and the fourth line (unlocked) “swings” through the ductal arch image (“swing technique”), providing an infinite number of cardiac planes in sequence. Each line generated the following plane(s): 1) Line 1: three-vessels and trachea view; 2) Line 2: five-chamber view and long axis view of the aorta (obtained by rotation of the five-chamber view on the y-axis); 3) Line 3: four-chamber view; and 4) “Swing” line: three-vessels and trachea view, five-chamber view and/or long axis view of the aorta, four-chamber view, and stomach. The algorithm was then tested in 50 normal hearts (15.3 – 40 weeks of gestation) and visualization rates for cardiac diagnostic planes were calculated. To determine if the algorithm could identify planes that departed from the normal images, we tested the algorithm in 5 cases with proven congenital heart defects.
In normal cases, the FAST Echo algorithm (3 locked lines and rotation of the five-chamber view on the y-axis) was able to generate the intended planes (longitudinal view of the ductal arch, pulmonary artery, three-vessels and trachea view, five-chamber view, long axis view of the aorta, four-chamber view): 1) individually in 100% of cases [except for the three-vessel and trachea view, which was seen in 98% (49/50)]; and 2) simultaneously in 98% (49/50). The “swing technique” was able to generate the three-vessels and trachea view, five-chamber view and/or long axis view of the aorta, four-chamber view, and stomach in 100% of normal cases. In the abnormal cases, the FAST Echo algorithm demonstrated the cardiac defects and displayed views that deviated from what was expected from the examination of normal hearts. The “swing technique” was useful in demonstrating the specific diagnosis due to visualization of an infinite number of cardiac planes in sequence.
This novel and simple algorithm can be used to visualize standard fetal echocardiographic planes in normal fetal hearts. The FAST Echo algorithm may simplify examination of the fetal heart and could reduce operator dependency. Using this algorithm, the inability to obtain expected views or the appearance of abnormal views in the generated planes should raise the index of suspicion for congenital heart disease.
STIC; prenatal diagnosis; congenital heart disease; ultrasound; four-dimensional; fetal heart
Atrial septal aneurysm is an uncommon condition. Between 1981 and 1984 10 cases of atrial septal aneurysm were diagnosed by real time cross sectional echocardiography performed in 4840 patients. The aneurysm was associated either with mitral valve prolapse (three patients) or with atrial septal defect (three patients) or occurred in isolation (four patients, two of whom had had a previous embolic event leading to the diagnosis of atrial septal aneurysm by cross sectional echocardiography). During cross sectional echocardiography the aneurysm appeared as a localised bulging of the interatrial septum, which was best seen in the subcostal four chamber view and in the parasternal short axis view at the level of the aortic root. The aneurysm either protruded into only the right atrium (five patients) or moved backwards and forwards between the right and the left atria during the cardiac cycle (five patients). This motion pattern might be related to changes in the interatrial pressure gradient. The two patients who had had a systemic embolism were given anticoagulant treatment, but none underwent surgery. It is concluded that the true prevalence of atrial septal aneurysm might have been underestimated before the routine use of cross sectional echocardiography, that cross sectional echocardiography enables definitive diagnosis of this condition by a non-invasive technique, and that an atrial septal aneurysm should be suspected and looked for by cross sectional echocardiography after an unexplained systemic embolism.
Thirty female dromedary camels were inseminated on a total of 50 occasions with 2-4 ml of fresh guanaco semen diluted with an equal volume of commercially available camel semen extender. Similarly, nine female guanacos were inseminated on 34 occasions with 4-6 ml of fresh, diluted camel semen. Only two of the dromedary females conceived; one aborted a female foetus on day 260 of gestation and the other gave birth to a stillborn female calf on day 365. Six conceptions occurred in the female guanacos. Two of these conceptuses, diagnosed by ultrasound, were resorbed between days 25 and 40 of gestation, one female foetus was aborted on day 291, another female foetus was aborted on day 302, and one female calf was stillborn on day 365 of gestation. The sixth foetus, a male, was born prematurely but alive after a 328-day gestation. It had a phenotypic appearance intermediate between that of a camel and a guanaco and its hybrid parentage was confirmed by the DNA fingerprinting of eight llama microsatellites. To our knowledge, this is the first viable hybrid ever to be produced between Old World and New World camelids, which have been reproductively isolated from one another for at least 11 million years. The preponderance of female hybrids is in accordance with Haldane's law. Histological examination of their ovaries revealed a failure of meiosis, with only an occasional abnormal oocyte surrounded by follicle cells. Although the diploid chromosone number of camels and guanacos is the same (2n = 74), sufficient genetic change has taken place to make the pairing of homologous chromosomes no longer possible.
Despite its economical, cultural, and biological importance, there has not been a large scale sequencing project to date for Camelus dromedarius. With the goal of sequencing complete DNA of the organism, we first established and sequenced camel EST libraries, generating 70,272 reads. Following trimming, chimera check, repeat masking, cluster and assembly, we obtained 23,602 putative gene sequences, out of which over 4,500 potentially novel or fast evolving gene sequences do not carry any homology to other available genomes. Functional annotation of sequences with similarities in nucleotide and protein databases has been obtained using Gene Ontology classification. Comparison to available full length cDNA sequences and Open Reading Frame (ORF) analysis of camel sequences that exhibit homology to known genes show more than 80% of the contigs with an ORF>300 bp and ∼40% hits extending to the start codons of full length cDNAs suggesting successful characterization of camel genes. Similarity analyses are done separately for different organisms including human, mouse, bovine, and rat. Accompanying web portal, CAGBASE (http://camel.kacst.edu.sa/), hosts a relational database containing annotated EST sequences and analysis tools with possibility to add sequences from public domain. We anticipate our results to provide a home base for genomic studies of camel and other comparative studies enabling a starting point for whole genome sequencing of the organism.
Heat shock protein 70 (Hsp70) is a molecular chaperone providing tolerance to heat and other challenges at the cellular and organismal levels. We sequenced a genomic cluster containing three hsp70 family genes linked with major histocompatibility complex (MHC) class III region from an extremely heat tolerant animal, camel (Camelus dromedarius). Two hsp70 family genes comprising the cluster contain heat shock elements (HSEs), while the third gene lacks HSEs and should not be induced by heat shock. Comparison of the camel hsp70 cluster with the corresponding regions from several mammalian species revealed similar organization of genes forming the cluster. Specifically, the two heat inducible hsp70 genes are arranged in tandem, while the third constitutively expressed hsp70 family member is present in inverted orientation. Comparison of regulatory regions of hsp70 genes from camel and other mammals demonstrates that transcription factor matches with highest significance are located in the highly conserved 250-bp upstream region and correspond to HSEs followed by NF-Y and Sp1 binding sites. The high degree of sequence conservation leaves little room for putative camel-specific regulatory elements. Surprisingly, RT-PCR and 5′/3′-RACE analysis demonstrated that all three hsp70 genes are expressed in camel's muscle and blood cells not only after heat shock, but under normal physiological conditions as well, and may account for tolerance of camel cells to extreme environmental conditions. A high degree of evolutionary conservation observed for the hsp70 cluster always linked with MHC locus in mammals suggests an important role of such organization for coordinated functioning of these vital genes.
Superoxide dismutase (SOD) is the first line of defense against oxidative stress induced by endogenous and/or exogenous factors and thus helps in maintaining the cellular integrity. Its activity is related to many diseases; so, it is of importance to study the structure and expression of SOD gene in an animal naturally exposed most of its life to the direct sunlight as a cause of oxidative stress. Arabian camel (one humped camel, Camelus dromedarius) is adapted to the widely varying desert climatic conditions that extremely changes during daily life in the Arabian Gulf. Studying the cSOD1 in C. dromedarius could help understand the impact of exposure to direct sunlight and desert life on the health status of such mammal. The full coding region of a putative CuZnSOD gene of C. dromedarius (cSOD1) was amplified by reverse transcription PCR and cloned for the first time (gene bank accession number for nucleotides and amino acids are JF758876 and AEF32527, respectively). The cDNA sequencing revealed an open reading frame of 459 nucleotides encoding a protein of 153 amino acids which is equal to the coding region of SOD1 gene and protein from many organisms. The calculated molecular weight and isoelectric point of cSOD1 was 15.7 kDa and 6.2, respectively. The level of expression of cSOD1 in different camel tissues (liver, kidney, spleen, lung and testis) was examined using Real Time-PCR. The highest level of cSOD1 transcript was found in the camel liver (represented as 100%) followed by testis (45%), kidney (13%), lung (11%) and spleen (10%), using 18S ribosomal subunit as endogenous control. The deduced amino acid sequence exhibited high similarity with Cebus apella (90%), Sus scrofa (88%), Cavia porcellus (88%), Mus musculus (88%), Macaca mulatta (87%), Pan troglodytes (87%), Homo sapiens (87%), Canis familiaris (86%), Bos taurus (86%), Pongo abelii (85%) and Equus caballus (82%). Phylogenetic analysis revealed that cSOD1 is grouped together with S. scrofa. The predicted 3D structure of cSOD1 showed high similarity with the human and bovine CuZnSOD homologues. The Root-mean-square deviation (rmsd) between cSOD1/hSOD1 and cSOD1/bSOD1 superimposed structure pairs were 0.557 and 0.425 A. The Q-score of cSOD1-hSOD1 and cSOD1-bSOD1 were 0.948 and 0.961, respectively.
CuZnSOD; 3D structure; cloning; molecular characterization; one-humped camel
OBJECTIVE--To test the hypothesis that isolated coarctation of the aorta is associated with relative hypoplasia of the mitral valve, even when the valve is morphologically normal. DESIGN--Cross sectional and Doppler echocardiography were used in a prospective, paired, case control study to compare mitral valve dimensions and diastolic transmitral flow characteristics as indices of left heart development. 40 children with isolated coarctation and 40 size matched controls were examined. Within the coarctation group 14 children with apical diastolic murmurs were compared with 14 size matched patients without murmurs. SETTING--A supraregional tertiary referral centre for paediatric cardiology. OUTCOME MEASURES--Mitral valve diameters, measured from the parasternal long axis, short axis, and apical four chamber views; mitral valve cross sectional area measured from the parasternal short axis view; peak early (E) and peak atrial (A) phase diastolic transmitral flow velocities measured by pulsed wave Doppler from the apical four chamber view; derived E/A ratio and pressure half time of decay from peak E. RESULTS--Mitral valve dimensions were significantly smaller in children with coarctation than in controls for long axis diameter (median 1.74 v 1.90cm, p = 0.0001), short axis diameter (2.21 v 2.28 cm, p = 0.027), and cross sectional area (2.37 v 3.15 cm2, p = 0.001). Peak E and A velocities were significantly higher in patients than in controls (0.9 v 0.82 ms-1, p = 0.013 and 0.61 v 0.51 ms-1, p = 0.007). The only difference between children with coarctation plus murmurs and those without murmurs was a marginally longer pressure half time. CONCLUSIONS--Smaller mitral valve dimensions and increased diastolic transmitral flow velocities in children with isolated coarctation compared with normal children suggests that coarctation may be part of a generalised hypoplasia of left heart structures.
Mycobacterium avium subspecies paratuberculosis (M. ap) is the causative agent of paratuberculosis or Johne's disease (JD) in herbivores with potential involvement in cases of Crohn's disease in humans. JD is spread worldwide and is economically important for both beef and dairy industries. Generally, pathogenic ovine strains (M. ap-S) are mainly found in sheep while bovine strains (M. ap-C) infect other ruminants (e.g. cattle, goat, deer), as well as sheep. In an effort to characterize this emerging infection in dromedary/Arabian camels, we successfully cultured M. ap from several samples collected from infected camels suffering from chronic, intermittent diarrhea suggestive of JD. Gene-based typing of isolates indicated that all isolates belong to sheep lineage of strains of M. ap (M. ap-S), suggesting a putative transmission from infected sheep herds. Screening sheep and goat herds associated with camels identified the circulation of this type in sheep but not goats. The current genome-wide analysis recognizes these camel isolates as a sub-lineage of the sheep strain with a significant number of single nucleotide polymorphisms (SNPs) between sheep and camel isolates (∼1000 SNPs). Such polymorphism could represent geographical differences among isolates or host adaptation of M. ap during camel infection. To our knowledge, this is the first attempt to examine the genomic basis of this emerging infection in camels with implications on the evolution of this important pathogen. The sequenced genomes of M. ap isolates from camels will further assist our efforts to understand JD pathogenesis and the dynamic of disease transmission across animal species.
Congenital aneurysms of the interventricular septum were found in a 29 year old man and his four year old son. Both were symptom free. In both, M mode and cross sectional echocardiography showed an aneurysm in the mid-muscular trabecular portion of the ventricular septum with considerable paradoxical motion of the aneurysmal segment. Otherwise the chamber dimensions, intracardiac structures, and cardiac function were normal for age. Congenital aneurysm of the interventricular septum is rare and these familial cases may be unique.
A nested polymerase chain reaction (nPCR)-based assay, was developed and evaluated for rapid detection of Trypanosoma evansi in experimentally infected mice and naturally infected camels (Camelus dromedarius). Four oligonucleotide primers (TE1, TE2, TE3 and TE4), selected from nuclear repetitive gene of T. evansi, were designed and used for PCR amplifications. The first amplification, using a pair of outer primers TE1 and TE2, produced a 821-bp primary PCR product from T. evansi DNA. The second amplification, using nested (internal) pair of primers TE3 and TE4, produced a 270-bp PCR product. T. evansi DNAs extracted from blood samples of experimentally infected mice and naturally infected Sudanese breed of dromedary camels were detected by this nested PCR-based assay. The nested primers TE3 and TE4 increased the sensitivity of the PCR assay and as little as 10 fg of T. evansi DNA (equivalent to a single copy of the putative gene of the parasite) was amplified and visualized onto ethidium bromide-stained agarose gels.
Amplification products were not detected when the PCR-based assay was applied to DNA from other blood parasites including Thieleria annulata, Babesia bigemina or nucleic acid free samples. Application of this nPCR-based assay to clinical samples resulted in direct detection of T. evansi from a variety of tissue samples collected from experimentally infected mice and blood from naturally infected camels. The described nPCR-based assay provides a valuable tool to study the epidemiology of T. evansi infection in camels and other susceptible animal populations.
The domesticated one-humped camel, Camelus dromedarius, is one of the most important animals in the Arabian Desert. It is exposed most of its life to both intrinsic and extrinsic genotoxic factors that are known to cause gross DNA alterations in many organisms. Ionic radiation and sunlight are known producers of Reactive Oxygen Species (ROS), one of the causes for DNA lesions. The damaged DNA is repaired by many enzymes, among of them Base Excision Repair enzymes, producing the highly mutagenic apurinic/apyrimidinicsites (AP sites). Therefore, recognition of AP sites is fundamental to cell/organism survival. In the present work, the full coding sequence of a putative cAPEX1 gene was amplified for the first time from C. dromedarius by RT-PCR and cloned (NCBI accession number are HM209828 and ADJ96599 for nucleotides and amino acids, respectively). cDNA sequencing was deduced to be 1041 nucleotides, of which 954 nucleotides encode a protein of 318 amino acids, similar to the coding region of the APEX1 gene and the protein from many other species. The calculated molecular weight and isoelectric point of cAPEX1 using Bioinformatics tools was 35.5 kDa and 8.11, respectively. The relative expressions of cAPEX1 in camel kidney, spleen, lung and testis were examined using qPCR and compared with that of the liver using a 18S ribosomal subunit as endogenous control. The highest level of cAPEX1 transcript was found in the testis; 325% higher than the liver, followed by spleen (87%), kidney (20%) and lung (5%), respectively. The cAPEX1 is 94%–97% similar to their mammalian counterparts. Phylogenetic analysis revealed that cAPEX1 is grouped together with that of S. scrofa. The predicted 3D structure of cAPEX1 has similar folds and topology with the human (hAPEX1). The root-mean-square deviation (rmsd) between cAPEX1 and hAPEX1 was 0.582 and the Q-score was 0.939.
Ape1/Ref-1/APEX1; 3D structure modeling; DNA repair; BER; cloning; molecular characterization; qPCR; one-humped camel
Dual chamber pacing improves functional status and reduces left ventricular outflow tract gradients in some, but not all patients with hypertrophic cardiomyopathy (HCM) by altering ventricular depolarisation. We investigated the use of biventricular (BIV) pacing in symptomatic patients with HCM.
8 patients aged 58±7yrs with symptomatic HCM underwent BIV pacing. 5 patients had LVOT gradients >30mmHg. Ventricular electrodes were placed in the right ventricle (RV) and a branch of the coronary sinus. An atrial electrode was inserted to achieve BIV pacing with a short AV delay. The short-term effects of different pacing modalities were assessed using 2-D and Doppler echocardiography. Symptoms and exercise tolerance were assessed after a month of each pacing mode. Long-term follow up data was available for 5 years.
Baseline EF was 67±14% and mean QRS duration was 132±26msecs. BIV pacing reduced QRS duration compared to RV pacing (129±46 vs. 205±54msecs, p<0.005). Five of the seven patients had baseline LVOT gradients (mean 67±25mmHg) that decreased to 41±15mm Hg with RV pacing (p<0.01) and 25±15mmHg with BIV pacing (p<0.005). Improvements in exercise time with active pacing occurred in six out of eight patients (75%), three (37.5%) had optimal exercise times with RV pacing and three with BIV pacing. Of the three patients with short term improvements with BIV pacing, one died 4 years post implant, one deteriorated with LV dilatation and one had the system explanted for infection.
BIV pacing showed short-term beneficial effects in some patients over and above RV pacing alone.
pacemakers; hypertrophic cardiomyopathy; conduction; biventricular
Four patients with primitive ventricle and normally related great vessels with stenotic subpulmonary outlet chamber (Holmes' heart with pulmonary stenosis) are reported. The history, physical examination, and chest x-ray film are not helpful in distinguishing Holmes' heart with pulmonary stenosis from tetralogy of Fallot. Electrocardiogram often provides the first clue to the presence of Holmes' heart; left axis deviation with or without left ventricular hypertrophy is an unusual finding in tetralogy of Fallot, but common in Holmes' heart. Selective ventriculography is diagnostic: the right ventricular outflow chamber overlies the aortic root and aortic valve in the frontal view in Holmes' heart with pulmonary stenosis, but is to the left of the aortic valve in tetralogy of Fallot; no ventricular septum can be identified in Holmes' heart. The diagnosis can be suspected in a child with clinical features of tetralogy of Fallot but atypical electrocardiogram, and can be established by angiography.
In human development, it is postulated based on histological sections, that the cardiogenic mesoderm rotates 180° with the pericardial cavity. This is also thought to be the case in mouse development where gene expression data suggests that the progenitors of the right ventricle and outflow tract invert their position with respect to the progenitors of the atria and left ventricle. However, the inversion in both cases is inferred and has never been shown directly. We have used 3D reconstructions and cell tracing in chick embryos to show that the cardiogenic mesoderm is organized such that lateralmost cells are incorporated into the cardiac inflow (atria and left ventricle) while medially placed cells are incorporated into the cardiac outflow (right ventricle and outflow tract). This happens because the cardiogenic mesoderm is inverted. The inversion is concomitant with movement of the anterior intestinal portal which rolls caudally to form the foregut pocket. The bilateral cranial cardiogenic fields fold medially and ventrally and fuse. After heart looping the seam made by ventral fusion will become the greater curvature of the heart loop. The caudal border of the cardiogenic mesoderm which ends up dorsally coincides with the inner curvature. Physical ablation of selected areas of the cardiogenic mesoderm based on this new fate map confirmed these results and, in addition, showed that the right and left atria arise from the right and left heart fields. The inversion and the new fate map account for several unexplained observations and provide a unified concept of heart fields and heart tube formation for avians and mammals.
Fate map; inversion; heart fields; heart tube; modeling; chick; 3D reconstruction; dye tracing; ablation
This study was conducted to underscore the spatial distribution of some biologically active proteins within the epididymal duct in the dromedary camel. Paraffin-embedded sections from different regions of epididymis were stained by conventional histological techniques and by immunohistochemistry. A battery of primary antibodies against six proteins (S100, alpha smooth muscle actin [α-SMA], connexin-43 [Cx43], galactosyltransferase [GalTase], angiotensin converting enzyme [ACE], and vascular endothelial growth factor [VEGF]) were used. The epididymal epithelium consisted of five cell populations: principal, basal, apical, dark, and halo cells. The histochemical findings indicated the absence of binding sites for VEGF and Cx43. The principal cells (PCs) showed variable immunoreactivity (IR) for ACE, S100, and GalTase throughout the whole length of the duct. The apical surfaces of most PCs (at the caput) and some PCs (at the corpus) exhibited intense ACE-IR, whereas those at the cauda displayed alternating negative and strong immunostaining. Similarly, moderate S100-IR was found in cytoplasm and nuclei of all PCs at the caput, few PCs at the corpus, and several PCs alternating with negative PCs at the cauda. In contrast, only some PCs showed weak to strong GalTase-IR in different regions. Apart from negative to weak positive S100-IR, basal cells failed to show IR for all other proteins. Apical cells displayed strong IR for ACE, S100, and GalTase with some regional differences. The peritubular and vascular smooth muscle cells revealed strong α-SMA-IR in all regions. In conclusion, the spatial distribution of different proteins in camel epididymis showed similarities and differences to other mammalian species. The region-specific topographic distribution of different proteins and cell types might indicate that the caput and cauda are metabolically more active than that of the corpus.
One-humped camel; Epididymis; Histology; Immunohistochemistry
Tomographic ultrasound imaging (TUI) is a new display modality that allows simultaneous visualization of up to eight parallel anatomical planes. This study was designed to determine the role of a novel algorithm combining spatiotemporal image correlation (STIC) and TUI to visualize standard fetal echocardiography planes.
Volume datasets from fetuses with and without congenital heart defects (CHD) were examined with a novel algorithm that allows simultaneous visualization of the three-vessel and trachea view, the four-chamber view, and outflow tracts. Visualization rates for these planes as well as ductal arch and five-chamber view were calculated.
1) 227 volume datasets from fetuses without (n=138) and with (n=14) CHD were reviewed; 2) among normal fetuses, the four-chamber view, five-chamber view, ductal arch, three-vessel and trachea view, left outflow tract and short axis of the aorta were visualized in 99% (193/195), 96.9% (189/195), 98.5 % (192/195), 88.2% (172/195), 93.3% (182/195), and 87.2% (170/195) of the volume datasets, respectively; 3) these views were visualized in 85% (17/20), 80% (16/20), 65% (13/20), 55% (11/20), 55% (11/20), and 70% (14/20) of the volume datasets, respectively, from fetuses with CHD; and 4) simultaneous visualization of the short axis of the aorta, three-vessel and trachea view, left outflow tract and four-chamber view was obtained in 78% (152/195) of the volume datasets from fetuses without CHD and in 40% (8/20) of those with CHD.
The three-vessel and trachea view, the four-chamber view, and both outflow tracts can be simultaneously visualized using a novel algorithm combining STIC and TUI.
Algorithm; 3D; 4D; three-dimensional; four-dimensional; STIC; spatiotemporal; congenital heart disease; spatiotemporal; prenatal diagnosis; fetal echocardiography
Allosuckling is a situation when a female nurses a non-filial offspring. It was described in various ungulate species; however for camels this is the first description of this behaviour. The aim of the study was to assess the occurrence of allosuckling in captive camels (Camelus bactrianus) and to test whether it can be explained as a ‘milk-theft’ (opportunistic behaviour of calves) or alternatively as an altruistic behaviour of females. During 2005 and 2007, nine camel females and ten calves in four zoological gardens in the Czech Republic were observed. In total, 373 sucking bouts were recorded, from which 32 were non-filial (the calf sucked from the non-maternal female). Allosuckling regularly appeared in captive camel herds. As predicted for the milk-theft explanation, the non-filial calves sucked more often in the lateral position and even did not suck in the antiparallel position at all. The non-filial calves preferably joined the filial calf when sucking but in five cases (15.6% of non-filial sucking bouts) the calves sucked from non-maternal dam without the presence of filial calf. We then expected the differences in terminations of sucking bouts by females but did not find any difference in sucking terminations for filial and non-filial calves. As the calves were getting older, the incidence of allosucking increased. This was probably because skills of the calf to outwit the non-maternal dam increased and/or the older calves might be more motivated for allosucking due to the weaning process. Finally, duration of a sucking bout was shorter with non-filial than filial calves. The results of the study support the hypothesis of ‘milk theft’, being mostly performed by calves behaving as opportunistic parasites, but we cannot reject certain level of altruism from the allonursing females or their increased degree of tolerance to non-filial calves.
Molecular mechanisms for the dorso-ventral patterning and interventricular septum formation in the embryonic heart are unknown. To investigate a role of Hand1/eHAND in cardiac chamber formation, we generated Hand1/eHAND knock-in mice where Hand1/eHAND cDNA was placed under the control of the MLC2V promoter. In Hand1/eHAND knock-in mice, the outer curvature of the right and left ventricles expanded more markedly. Moreover, there was no interventricular groove or septum formation, although molecularly, Hand1/eHAND knock-in hearts had two ventricles. However, the morphology of the inner curvature of the ventricles, the atrioventricular canal, and the outflow tract was not affected by Hand1/eHAND expression. Furthermore, expression of Hand1/eHAND in the whole ventricles altered the expression patterns of Chisel, ANF, and Hand2/dHAND but did not affect Tbx5 expression. In contrast, the interventricular septum formed normally in transgenic embryos overexpressing Hand1/eHAND in the right ventricle but not in the boundary region. These results suggested that Hand1/eHAND is involved in expansion of the ventricular walls and that absence of Hand1/eHAND expression in the boundary region between the right and left ventricles may be critical in the proper formation of the interventricular groove and septum. Furthermore, Hand1/eHAND is not a master regulatory gene that specifies the left ventricle myocyte lineage but may control the dorso-ventral patterning in concert with additional genes.