A comparison of the effects of carbon dioxide and medical air for abdominal insufflation on respiratory parameters in xylazine-sedated sheep undergoing laparoscopic artificial insemination

AIMS: To determine if abdominal insufflation with medical air will improve oxygenation and ventilation parameters when compared to insufflation with CO₂ in xylazine-sedated sheep undergoing laparoscopic artificial insemination (AI).

METHODS: Forty-seven sheep underwent oestrus synchronisation and were fasted for 24 hours prior to laparoscopic AI. Each animal was randomised to receive either CO₂ or medical air for abdominal insufflation. An auricular arterial catheter was placed and utilised for serial blood sampling. Respiratory rates (RR) and arterial blood samples were collected at baseline, after xylazine (0.1?mg/kg I/V) sedation, 2 minutes after Trendelenburg positioning, 5 minutes after abdominal insufflation, and 10 minutes after being returned to a standing position. Blood samples were collected in heparinised syringes, stored on ice, and analysed for arterial pH, partial pressure of arterial O₂ (PaO₂), and CO₂ (PaCO₂). The number of ewes conceiving to AI was also determined.

RESULTS: Repeated measures ANOVA demonstrated temporal effects on RR, PaO₂, PaCO₂ and arterial pH during the laparoscopic AI procedure (p<0.001), but no difference between insufflation groups (p>0.01). No sheep experienced hypercapnia (PaCO₂>50?mmHg) or acidaemia (pH<7.35). Hypoxaemia (PaO₂<70?mmHg) was diagnosed during the procedure in 14/22 (64%) ewes in the CO₂ group compared with 8/23 (35%) ewes in the medical air group (p=0.053). Overall, 15/20 (75%) ewes in the CO₂ group conceived to AI compared with 16/22 (72.7%) in the medical air group (p=0.867).

CONCLUSIONS AND CLINICAL RELEVANCE: There were no statistical or clinical differences in RR, PaO₂, PaCO₂, pH, or conception to AI when comparing the effects of CO₂ and medical air as abdominal insufflation gases. None of the sheep experienced hypercapnia or acidaemic, yet 42% (19/45) of sheep developed clinical hypoxaemia, with a higher percentage of ewes in the CO₂ group developing hypoxaemia than in the medical air group. Based on the overall analysis, medical air could be utilised as a comparable alternative for abdominal insufflation during laparoscopic AI procedures.     

Isolation,purification, and biochemical characterization of a novel water soluble protein from Inca peanut (Plukenetia volubilis L.)

A water soluble storage albumin from Inca peanut (IPA) accounted for approximately 25% (w/w) of defatted seed flour weight, representing 31% of the total seed protein. IPA is a 3S storage protein composed of two glycosylated polypeptides, with estimated molecular weights (MW) of 32800 and 34800 Da, respectively. IPA has an estimated sugar content of 4.8% +/- 0.92% (n = 6). IPA is a basic protein (pI of approximately 9.4) and contains all of the essential amino acids in adequate amounts when compared to the FAO/WHO recommended pattern for a human adult. The tryptophan content of IPA is unusually high (44 mg/g of protein), whereas the phenylalanine content is low (9 mg/g of protein). IPA is a highly digestible protein in vitro.     

Biochemical characterization of amandin,the major storage protein in almond (Prunus dulcis L.)

The almond major storage protein, amandin, was prepared by column chromatography (amandin-1), cryoprecipitation (amandin-2), and isoelectric precipitation (amandin-3) methods. Amandin is a legumin type protein characterized by a sedimentation value of 14S. Amandin is composed of two major types of polypeptides with estimated molecular weights of 42-46 and 20-22 kDa linked via disulfide bonds. Several additional minor polypeptides were also present in amandin. Amandin is a storage protein with an estimated molecular weight of 427,300 +/- 47,600 Da (n = 7) and a Stokes radius of 65.88 +/- 3.21 A (n = 7). Amandin is not a glycoprotein. Amandin-1, amandin-2, and amandin-3 are antigenically related and have similar biochemical properties. Amandin-3 is more negatively charged than either amandin-1 or amandin-2. Methionine is the first essential limiting amino acid in amandin followed by lysine and threonine.     

Electrophoretic and immunological analyses of almond (Prunusdulcis l.) genotypes and hybrids

Aqueous extracts from sixty almond samples representing various genotypes and interspecies hybrids of almond, including almond-peach, were analyzed for protein and peptide content using electrophoresis, Western immunoblotting, and enzyme-linked immunosorbent assay (ELISA). Nondenaturing nondissociating polyacrylamide gel electrophoresis (NDND-PAGE) of the aqueous extracts indicated that a single major storage protein (almond major protein -- AMP or amandin) dominated the total soluble protein composition. Denaturing SDS--PAGE analyses of the aqueous extracts revealed that the AMP was mainly composed of two sets of polypeptides with estimated molecular masses in the ranges of 38--41 kDa and 20--22 kDa, regardless of the source; however, distinct variations in the intensity and electrophoretic mobility of some bands were noted between samples. In addition to AMP, several minor polypeptides were also present in all the genotypes, and variations were seen in these as well. Regardless of the genotype, AMP was recognized in Western blots by rabbit polyclonal anti-AMP antibodies, mouse monoclonal anti-AMP antibodies (mAbs), and serum IgE from patients displaying strong serum anti-almond IgE reactivity. As with protein staining results, antibody reactivity also revealed common patterns but displayed some variation between samples. An anti-AMP inhibition ELISA was used to quantify and compare aqueous extracts for various samples. All samples (n = 60) reacted in this assay with a mean +/- standard deviation (sigma n) = 0.82 +/- 0.18 when compared to reference aqueous extract from Nonpareil designated as 1.0.     

Biochemical and spectroscopic characterization of almond and cashew nut seed 11S legumins, amandin and anacardein

Native, undenatured amandin and anacardein secondary structures were estimated to be, respectively, 56.4 and 49% β-sheet, 14 and 23.7% α-helix, and 29.6 and 27.4% random coil. Circular dichroic (CD) and fluorescence spectroscopy were used to assess structural changes in amandin and anacardein subjected to denaturing treatments that included heat (100 °C, 5 min), guanidium HCl (GuHCl), urea, sodium dodecyl sulfate (SDS), and reducing agent, 2% v/v β-mercaptoethanol (βME) + heat. Mouse monoclonal antibodies (mAbs) 4C10 and 4F10 directed against amandin and 1F5 and 4C3 directed against anacardein were used to assess the influence of denaturing treatments on the immunoreactivity of amandin and anacardein. Among the denaturing treatments investigated, SDS and β-ME caused a significant reduction in the immunoreactivity of amandin and anacardein when probed with mAb 4C10 and 4C3, respectively.     

Detection and stability of the major almond allergen in foods

Almond major protein (AMP or amandin), the primary storage protein in almonds, is the major allergen recognized by almond-allergic patients. A rabbit antibody-based inhibition ELISA assay for detecting and quantifying AMP in commercial foods has been developed, and this assay, in conjunction with Western blotting analyses, has been applied to the investigation of the antigenic stability of AMP to harsh food-processing conditions. The ELISA assay detects purified AMP at levels as low as 87 +/-16 ng/mL and can detect almond at between 5 and 37 ppm in the tested foods. The assay was used to quantify AMP in aqueous extracts of various foods that were defatted and spiked with known amounts of purified AMP or almond flour. In addition, AMP was quantified in commercially prepared and processed almond-containing foods. Neither blanching, roasting, nor autoclaving of almonds markedly decreased the detectability of AMP in subsequent aqueous extracts of almonds. Western blots using both rabbit antisera and sera from human almond-allergic patients confirm a general stability of the various peptides that comprise this complex molecule and show that the rabbit antibody-based assay recognizes substantially the same set of peptides as does the IgE in sera from almond-allergic patients.