Search

Baboon models are often used to investigate haemostatic diseases, such as acquired thrombotic thrombocytopenic purpura or bacterial sepsis-induced disseminated intravascular coagulation, and their potential treatment with novel drugs. Thrombin generation is vital for these models, and an important potential therapeutic target. We investigated the thrombin generation profile of the Chacma baboon (Papio ursinus - a common pre-clinical model) including the effects of sex and ABO blood group. Thrombin generation curves, lag times, peak heights, times-to-peak, velocity indexes and Endogenous Thrombin Potentials (ETPs) of 40 adult Chacma baboons were assessed and compared with normal human plasma, using a low concentration of tissue factor (1 pM) and phospholipids. Reference intervals were calculated, and results compared between O and non-O ABO blood groups, and between males and females. Lag times of all baboons fell within the human reference interval. Most animals (n = 32; 80%) had times-to-peak above, and velocity indexes and peak heights markedly below (n = 27; 68%) the human range. However, 97.5% of baboons had an ETP above the human reference interval, indicating greater overall thrombin generation. ABO blood group had no effect, but males (n = 14; 35%) had less potent thrombin generation than females (n = 26; 65%), with significantly longer lag times (p = 0.0475), lower peak thrombin concentrations (p = 0.0203), and lower ETPs (p = 0.0238). Chacma baboons have greater overall endogenous thrombin generation potentials than humans, which is even more prominent in females. This should be considered when designing future baboon model experiments involving the haemostatic system, or when evaluating novel therapies in these animals., (© 2023. The Author(s).)

Thrombosis is a potentially life-threatening condition related to roughly a quarter of all deaths globally. Many of these deaths occur inside healthcare facilities due to possibly preventable causes. Therefore, understanding the etiological factors involved in excessive thrombosis may significantly contribute to the successful identification, management and education of people who have an increased risk of thrombosis. We performed a retrospective file-audit of all forensic autopsy reports conducted at the Free State Forensic Pathology Mortuary in Bloemfontein, South Africa, over 10 years. We collected the age at death, gender and ethnicity of each person included in the study. The presence and location of the thrombosis and any underlying disorder or disease were noted. The overall prevalence of thrombosis for the total study population was 0.97%. Pulmonary embolisms (PE's) were much more common than coronary thromboses. Most PE's had known contributory risk factors, where coronary thrombosis-related deaths occurred suddenly without known risk factors. Bone fractures were the most prominent risk factor associated with PE's. Females of African descent had a consistently high prevalence of thrombosis after the age of 30 years. Males of European descent showed an unexpected peak in prevalence during the 4th decade. Since most deaths occurred in patients with conditions known to contribute to venous thrombosis, we conclude that intensified public awareness efforts are required in our region to assist the general public in identifying risk factors for thrombosis, thereby decreasing the burden this potentially preventable disorder places on society., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

TTP is a life-threatening disorder with limited pharmaceutical treatment options. Recently, the potential of streptokinase in the treatment of acquired TTP was demonstrated in humans in vitro, and in vivo in a mouse model. We aimed to determine the in vitro and in vivo effects of streptokinase in an established Papio ursinus model of acquired TTP. In vitro: VWF activities & multimer patterns and thromboelastograms were assessed with increasing concentrations of streptokinase. In vivo: After induction of TTP, escalating streptokinase doses (ranging from 50,000 to 900,000 IU) were administered, and the effects of streptokinase assessed on peripheral blood counts, fibrinolysis, VWF activities & multimer patterns and thromboelastograms. In an extension of the study, high-dose streptokinase (1,500,000-3,000,000 IU) was administered to another baboon. After spiking, fibrinolysis with loss of large VWF multimers was observed at [2200 IU/mL]-roughly equivalent to 1,500,000 IU. However, administration of escalating intravenous streptokinase doses had no in vivo effect on the TTP phenotype, and in vivo increases in plasmin activity were mild when compared with baseline, even at high doses. Minimal effect on VWF multimer patterns was observed but only at doses ≥ 1500,000 IU. Streptokinase is not effective in resolving TTP in a Papio ursinus model of TTP, possibly due to limited activation of the baboon fibrinolytic system. Modifications to this model, the use of alternative higher animal models, or alternative thrombolytics, should be considered to establish proof-of-concept., (© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Background: Human Leucocyte Antigens (HLA) play a vital role in disease pathogenesis and transplant rejection. HLA-typing is a useful tool in predicting disease progression and to identify potential organ donors. Due to human migration and known ethnic variation, frequent targeted HLA sequencing of specific populations is crucial to increase their representation in global reference panels., Materials and Methods: We performed a retrospective file audit of all HLA-typings done in our setting from 2005-2019. We discuss data for the major HLA-A, HLA-B, HLA-C, and HLA-DRB1 allele groups., Results: Overall, the most common allele groups were HLA-A∗02, HLA-B∗15, HLA-C∗07 and HLA-DRB1∗03. For the African descent group, the most common alleles were HLA-A∗30, HLA-B∗15, HLA-C∗07 and HLA-DRB1∗03. For the European descent group, the most common alleles were HLA-A∗02, HLA-B∗07, HLA-C∗07 and HLA-DRB1∗15. For the mixed ancestry group, the most common allele groups were HLA-A∗02, HLA-B∗15, HLA-C∗02 and HLA-DRB1∗13. HLA-B∗44 was identified as the most common allele group in patients with renal failure., Discussion and Conclusion: The significant variation within the HLA frequencies between the different ethnic groups highlights the value of population-specific HLA-typing. Furthermore, the identification of HLA-B∗44 as a prominent HLA in our renal failure population warrants in-depth investigation of this group., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Published by Elsevier Ltd.)

Taro [ Colocasia esculenta (L.) Schott] has the potential to address food and nutrition insecurity in sub-Saharan Africa. However, the nutrient content of taro is yet to be fully elucidated. The objective of this study was to evaluate mineral element content as a proxy for nutritional value of different taro genotypes. The study evaluated 14 taro accessions at Roodeplaat and Umbumbulu in South Africa based on their calcium (Ca), iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), sodium (Na), phosphorous (P) and zinc (Zn) content. The accessions were planted in a randomized complete block design, replicated three times under field conditions. The mineral element content varied significantly (p < 0.05) among the genotypes. Genotypes Amad7-2, Umbu8 and Amad101 exhibited high Ca (≥432 mg kg -1 ), Fe (≥32 mg kg -1 ) and Mg (≥229 mg kg -1 ) across the locations. The first principal component (PC) accounted for 33.7% of the variation and was strongly associated with Zn (r = 0.94, p < 0.001) and P (r = 0.89, p < 0.001). The second PC explained 29.7% of the variation and was associated with Na (r = 0.83, p < 0.001), Mg (r = 0.76, p < 0.001) and K (r = 0.55, p < 0.05). Fe and Mn contributed below the 12.5% threshold to the PCs and were considered as less discriminatory among the accessions. The negative correlations among some of the mineral elements would be a challenge for selection and breeding of nutritious taro accessions. This information is essential to select superior local accessions based on their mineral element content for developing breeding populations and lines for improving nutrition quality among poor households in sub-Saharan Africa., Competing Interests: The authors declare no conflict of interest., (© 2021 Published by Elsevier Ltd.)

The study was conducted to determine the influence of male-male competition on reproductive performance and male mortality of Cobb 500 broiler breeder flocks following double intra-spiking. Broiler breeders were housed in 3 open-sided houses each accommodating 8,200 females and 820 males. Males of the same age on the same farm were exchanged between the houses (intra-spiking) to stimulate competition thereby changing the social hierarchy of each house. Intra-spiking was performed by replacing 25, 35, and 45% of males between the houses at 40 and 48 weeks of age (WOA), respectively. Eggs were collected from 36 to 55 WOA, when egg fertility and male mortality were recorded. Data was analyzed using repeated measures techniques of SAS 9.4, modeling the covariance structure of the observed data. Male-male competition (intra-spiking), age and their interaction significantly (P < 0.05) influenced egg fertility, hatchability, and male mortality. Average fertility and hatchability were increased in the 45% intra-spiked flocks (P < 0.05) (95.89 and 85.83%) compared with the 35% (95.13 and 86.30%) and 25% (94.42 and 0.23%) intra-spiked flocks. Fertility and hatchability with the 45% double intra-spiked flocks was consistently higher (P < 0.05) over time than the other double intra-spiked flocks. Male mortality was lower (P < 0.05) in the 45% intra-spiked flock (0.23%) than in the 35% (0.40%) and 25% (0.44%) intra-spiked flocks. After double intra-spiking, the male mortality in the 25 and 35% double intra-spiked flocks significantly increased (P < 0.05), whereas that of 45% intra-spiked flocks remained relatively low. Male mortality in the 45% intra-spiked flocks was consistently low over time than other double intra-spike levels from 45 WOA until the end of the trial. Noteworthy, egg fertility and hatchability gradually decreased, and mortality increased with increasing flock age toward the end of the productive life cycle. High level of male-male competition (45%) showed great promise as a tool to slow down the decrease in egg fertility and hatchability, and reduce male mortality in aging broiler breeder flocks., (© 2019 Poultry Science Association Inc.)

Background: Von Willebrand factor (VWF) has a central role in primary haemostasis and is a popular pharmaceutical target in the prevention and treatment of disorders such as acute coronary syndrome and thrombotic thrombocytopenic purpura. We have evaluated numerous possible treatments targeting VWF in the chacma baboon. Unfortunately, no data exist regarding the molecular similarity of the chacma baboon and human VWF protein, resulting in limited translatability of results., Methods: Sanger sequencing was performed of the functionally vital VWF exon 28. The sequences were then compared to the human reference sequence., Results: The baboon and human VWF amino acid sequences were 99.1% similar, with only 4 radical amino acid changes found. No radical amino acid changes were found within the functionally vital areas of the amino acid sequence., Conclusion: The chacma baboon VWF is similar enough to the human to produce reliable and translatable pre-clinical results with human-targeted anti-VWF agents., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Staphylococcus aureus infections can produce systemic bacteremia and inflammation in humans, which may progress to severe sepsis or septic shock, even with appropriate antibiotic treatment. Sepsis may be associated with disseminated intravascular coagulation and consumptive coagulopathy. In some types of mouse infection models, the plasma coagulation protein factor XI (FXI) contributes to the pathogenesis of sepsis. We hypothesize that FXI also contributes to the pathogenesis of sepsis in primates, and that pharmacological interference with FXI will alter the outcome of Staphylococcus aureus -induced lethality in a baboon model. Pretreatment of baboons with the anti-FXI antibody 3G3, a humanized variant of the murine monoclonal 14E11 that blocks FXI activation by FXIIa, substantially reduced the activation of coagulation, as reflected by clotting times and plasma complexes of coagulation proteases (FXIIa, FXIa, FIXa, FXa, FVIIa, and thrombin) with serpins (antithrombin or C1 inhibitor) following infusion of heat-inactivated S aureus 3G3 treatment reduced fibrinogen and platelet consumption, fibrin deposition in tissues, neutrophil activation and accumulation in tissues, cytokine production, kininogen cleavage, cell death, and complement activation. Overall, 3G3 infusion protected the structure and function of multiple vital organs, including lung, heart, liver, and kidney. All treated animals reached the end point survival (7 days), whereas all nontreated animals developed terminal organ failure within 28 hours. We conclude that FXI plays a role in the pathogenesis of S aureus -induced disseminated intravascular coagulation and lethality in baboons. The results provide proof of concept for future therapeutic interventions that may prevent sepsis-induced organ failure and save lives in certain forms of sepsis., (© 2019 by The American Society of Hematology.)

Background . Cardiovascular disease is a major cause of deaths. Elevated cholesterol levels to above the normal reference range is a major risk factor for developing cardiovascular disease. Current guidelines recommend the use of cholesterol-lowering drugs to lower cholesterol levels to within the normal reference range. However, the American Heart Association further recommends a change in lifestyle in managing cholesterol levels. Thus, cholesterol-lowering drugs may not be needed if a lifestyle-change alone is sufficient in lowering cholesterol levels to within normal ranges. Unfortunately, limited examples exist in academic literature to illustrate the effectiveness of lifestyle change alone in lowering of cholesterol levels. Case report . We report a case of a 33-year-old man, with moderately elevated cholesterol levels and a family history of cardiovascular disease. Method . The man followed an altered healthy fat diet accompanied with moderate exercise for 6 weeks, without the addition of cholesterol-lowering agents. Results . At the 6-week follow-up, he was able to decrease his total cholesterol by 40.25% and low-density lipid cholesterol by 52.8%, to within normal ranges. The cholesterol levels remained within normal ranges after 6 months. Conclusion . This case illustrates that in some individuals, lifestyle change alone is sufficient to lower moderately elevated cholesterol levels., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

BACKGROUND Platelet reactivity assessment is an important tool in both the causal determination of bleeding diathesis as well as in the evaluation of the efficacy of anti-platelet therapy in patients at risk of thrombosis. Sodium citrate is the most widely used anticoagulant for hemostasis investigations. However, some doubt exists over the suitability of sodium citrate in platelet function testing. Hirudin has been suggested as a superior replacement. Nevertheless, only 1 study compared citrated and hirudin treated samples with light transmission aggregometry. Therefore, limited evidence exists to conclusively prove the supremacy of hirudin over sodium citrate in light transmission aggregometry. The aim of our study was to compare citrated and hirudin treated samples, collected in commercially available blood collection tubes, using the 5 most common agonists, with light transmission aggregometry. MATERIAL AND METHODS Blood was obtained from 20 healthy volunteers. Platelet counts were performed on platelet-rich plasma. Light transmission aggregometry was performed within 4 h of sample collection using ADP, collagen, arachidonic acid, epinephrine, and ristocetin as agonists. RESULTS Platelet counts for the respective anticoagulants did not differ significantly. ADP-induced aggregation was comparable between the samples. However, among all the agonists, hirudin-treated platelets had significantly weaker aggregatory responses. CONCLUSIONS Commercially available sodium citrate should remain the anticoagulant of choice for routine platelet function testing in our setting. However, the time limitation associated with the use of sodium citrate in platelet function testing remains a concern. Thus, alternative anticoagulants should still be explored.

Thrombotic thrombocytopenic purpura (TTP) is a microangiopathic disorder diagnosed by thrombocytopenia and hemolytic anemia, associated with a deficiency in von Willebrand factor (VWF)-cleaving protease ADAMTS13. Current treatment is based on plasma infusion for congenital TTP, or plasma exchange, often in combination with immunosuppressive agents, for acquired TTP. These treatment methods are not always effective; therefore, new treatment methods are highly necessary. N -acetylcysteine (NAC), an FDA-approved anti-mucolytic agent, is a possible new treatment strategy for TTP, as it was demonstrated to reduce disulfide bonds in VWF, thereby decreasing VWF multimers size and hence their prothrombotic potential. We investigated whether NAC, without concurrent plasma exchange and immunosuppressive therapy, is effective in preventing and resolving TTP signs, using well-established murine and baboon models for TTP. In mice, prophylactic administration of NAC was effective in preventing severe TTP signs. This in vivo finding was supported by in vitro data demonstrating the VWF multimer-reducing properties of NAC in solution. Nonetheless, in both mice and baboons, administration of NAC was not effective in resolving preexisting TTP signs; thrombocytopenia, hemolytic anemia, and organ damage could not be reversed, as thrombus resolution was not achieved. Failure to improve clinical outcome occurred even though a reduction in VWF multimers was observed, demonstrating that NAC was efficient in reducing disulfide bonds in circulating VWF multimers. In conclusion, prophylactic administration of NAC, without concurrent plasma exchange, was effective in preventing severe TTP signs in mice, but NAC was not effective in resolving preexistent acute TTP signs in mice and baboons., (© 2017 by The American Society of Hematology.)

Taro (Colocasia esculenta (L.) Schott) is widely distributed in tropical and sub-tropical areas. However, its origin, diversification and dispersal remain unclear. While taro genetic diversity has been documented at the country and regional levels in Asia and the Pacific, few reports are available from Americas and Africa where it has been introduced through human migrations. We used eleven microsatellite markers to investigate the diversity and diversification of taro accessions from nineteen countries in Asia, the Pacific, Africa and America. The highest genetic diversity and number of private alleles were observed in Asian accessions, mainly from India. While taro has been diversified in Asia and the Pacific mostly via sexual reproduction, clonal reproduction with mutation appeared predominant in African and American countries investigated. Bayesian clustering revealed a first genetic group of diploids from the Asia-Pacific region and to a second diploid-triploid group mainly from India. Admixed cultivars between the two genetic pools were also found. In West Africa, most cultivars were found to have originated from India. Only one multi-locus lineage was assigned to the Asian pool, while cultivars in Madagascar originated from India and Indonesia. The South African cultivars shared lineages with Japan. The Caribbean Islands cultivars were found to have originated from the Pacific, while in Costa Rica they were from India or admixed between Indian and Asian groups. Taro dispersal in the different areas of Africa and America is thus discussed in the light of available records of voyages and settlements.

Anti-platelet agents play a central part in the treatment and prevention of acute thrombotic events. Discriminating animal models are needed for the development of novel agents. The chacma baboon has been extensively used as a model to evaluate anti-platelet agents. However, limited data exist to prove the translatability of this species to humans. We aimed to determine the suitability of the chacma baboon in preclinical human targeted GPIIb/IIIa, GPIbα and P2Y12 studies. Light-transmission platelet aggregometry (LTA), whole blood impedance aggregometry, receptor number quantification and genomic DNA sequencing were performed. Baboon ADP and arachidonic acid-induced LTA aggregation results differed significantly from human values, even at increased concentrations. LTA ristocetin-induced agglutination was comparable between species, but baboon platelets needed twice the concentration of ristocetin to elicit a similar response. Citrated baboon blood had significantly less aggregation than humans when evaluated with impedance aggregometry. However, hirudinised baboon whole blood gave similar aggregation as humans at the same agonist concentrations. GPIIb, GPIIIa and GPIbα numbers were significantly more on the baboon platelets. None of the amino acids deemed vital for receptor function, ligand binding or receptor inhibition, were radically different between the species. However, a conservative change in a calcium-binding region of GPIIb may render the baboon platelets more sensitive to calcium-binding agents. The chacma baboon may be used for the evaluation of human-targeted GPIIb/IIIa-, GPIbα- and P2Y12-inhibiting agents. However, the best anticoagulant, optimal agonist concentrations, increase in receptor number and sequence differences must be considered for any future studies.

An effective and safe anti-platelet drug is central to the management of patients with acute coronary syndrome (ACS). Glycoprotein VI (GPVI) is currently regarded as a potential target for novel anti-platelet agents due to its collagen-binding potential. Development of anti-thrombotics is associated with testing in animals. We have previously successfully evaluated anti-platelet drugs in the Cape Chacma baboon (Papio ursinus). However, various anti-GPVI agents did not have an effect on baboons when evaluated in our arterial thrombosis model. To evaluate the suitability of baboons for GPVI studies, we performed collagen-induced platelet aggregation, GPVI quantification and DNA sequencing. Baboon platelets needed double the amount of collagen compared to human platelets to illicit proper aggregation. GPVI quantification was unsuccessful due to non-binding of monoclonal antibodies. Sequencing of the GPVI gene revealed 36 SNPs leading to 14 amino acid changes, as well as a 9 bp deletion causing a 3 amino acid deletion. Several of the amino acid changes were within the ligand binding region of GPVI, causing limited binding of humanized anti-GPVI antibodies to the baboon platelets. Therefore, the baboon was deemed not suitable to evaluate human targeted anti-GPVI agents.

Background: The perishability of Brassica chinensis poses a major challenge to distribution and marketing. The aim of this work was to select a suitable modified atmosphere packaging to retain the overall quality and bioactive compounds during storage., Results: Four types of biorientated polypropylene packaging (BOPP)--BOPP03, BOPP04, BOPP05 and BOPP06--with different perforations were evaluated regarding the maintenance of quality parameters (weight loss, leaf yellowing, colour L*, C*, h°), decay, chlorophyll a, chlorophyll b, bioactive compounds (carotenoids, ascorbic acid, total phenolic compounds), antioxidant scavenging activity, overall appearance and odour evaluation, at 10°C at 2, 4, 6, 8 and 10 days. Leaves were packed in BOPP (two 2-mm holes) and packed and unpacked leaves were included for comparison. The modified atmosphere created (2% O2 and 7% CO2) inside the BOPP05 reduced leaf yellowing (higher h°), improved the overall appearance with acceptable odour, moderately maintained chlorophyll a and b, bioactive compounds and antioxidant scavenging activity, and remained marketable for up to 10 days at 10°C. Gas composition within the packages influenced the retention of bioactive compounds and overall quality., Conclusion: Application of BOPP05 is a promising method for extending the shelf life of B. chinensis leaves in order to promote its utilisation and commercialisation via urban fresh-produce markets., (© 2012 Society of Chemical Industry.)

To prevent thrombosis in high-risk acute coronary syndrome patients undergoing percutaneous coronary intervention for re-vascularisation, concomitant administration of a glycoprotein IIb/IIIa inhibitor, such as abciximab, tirofiban or eptifibatide, is recommended. Abciximab and eptifibatide are mostly preferred over tirofiban, which is less effective in preventing ischaemic events. We compared the efficacy and bleeding potential of escalating doses of tirofiban and abciximab in non-human primates. The efficacy of tirofiban and abciximab in inhibiting cyclic flow reductions (CFRs) was tested in a high shear arterial thrombosis model. Bleeding was evaluated with the template bleeding time and an incision bleeding model. Abciximab completely inhibited arterial thrombosis after injection of its therapeutic bolus dose. With tirofiban, a dose three times higher than the recommended therapeutic dose caused weak inhibition characterised by a return of CFRs after re-injury. At nine times the recommended therapeutic dose, complete inhibition was observed, and the efficacy of tirofiban was comparable to abciximab at its therapeutic bolus dose. Blood loss was less than with abciximab at its effective dose. In this model, tirofiban compared favourably with abciximab, although only at a dose of 3-9 times the therapeutic dose, and caused less bleeding than abciximab.

The pathophysiology of thrombotic thrombocytopenic purpura (TTP) can be explained by the absence of active ADAMTS13, leading to ultra-large von Willebrand factor (UL-VWF) multimers spontaneously interacting with platelets. Preventing the formation of UL-VWF-platelet aggregates therefore is an attractive new treatment strategy. Here, we demonstrate that simultaneous administration of the inhibitory anti-VWF monoclonal antibody GBR600 and the inhibitory anti-ADAMTS13 antibody 3H9 to baboons (prevention group) precluded TTP onset as severe thrombocytopenia and hemolytic anemia were absent in these animals. In addition, partial VWF inhibition was not enough to prevent thrombocytopenia, demonstrating the specificity of this therapeutic strategy. GBR600 treatment of baboons during acute TTP (treatment group) resulted in a rapid recovery of severe thrombocytopenia similar to the platelet count increases observed in TTP patients treated by plasma exchange. Baboons in the control group only injected with 3H9 developed early stages of TTP as previously described. Hence, inhibiting VWF-GPIb interactions is an effective way to prevent and treat the early symptoms of acquired TTP in baboons.

ALX-0681 is a therapeutic Nanobody targeting the A1-domain of VWF. It inhibits the interaction between ultra-large VWF and platelet GpIb-IX-V, which plays a crucial role in the pathogenesis of thrombotic thrombocytopenic purpura (TTP). In the present study, we report the efficacy and safety profile of ALX-0681 in a baboon model of acquired TTP. In this model, acute episodes of TTP are induced by administration of an ADAMTS13-inhibiting mAb. ALX-0681 completely prevented the rapid onset of severe thrombocytopenia and schistocytic hemolytic anemia. After induction of TTP, platelet counts also rapidly recovered on administration of ALX-0681. This effect was corroborated by the full neutralization of VWF activity. The schistocytic hemolytic anemia was also halted and partially reversed by ALX-0681 treatment. Brain CT scans and post mortem analysis did not reveal any sign of bleeding, suggesting that complete neutralization of VWF by ALX-0681 under conditions of thrombocytopenia was not linked with an excessive bleeding risk. The results obtained in this study demonstrate that ALX-0681 can successfully treat and prevent the most important hallmarks of acquired TTP without evidence of a severe bleeding risk. Therefore, ALX-0681 offers an attractive new therapeutic option for acquired TTP in the clinical setting.

Background: Polyunsaturated fatty acids (PUFAs) have antifungal properties, but the mode by which they induce their action is not always clear. The aim of the study was to investigate apoptosis as a mode of action of antifungal PUFAs (stearidonic acid, eicosapentaenoic acid and docosapentaenoic acid) which are inhibitory towards biofilm formation of C. albicans and C. dubliniensis., Methods: Candida biofilms were grown in the absence or presence of 1mM PUFAs (linoleic acid, stearidonic acid, eicosapentaenoic acid, docosapentaenoic acid) for 48h at 37°C. The effect of these PUFAs on the membrane fatty acid profile and unsaturation index, oxidative stress, mitochondrial transmembrane potential and apoptosis was evaluated., Results: When biofilms of C. albicans and C. dubliniensis were exposed to certain PUFAs there was an increase in unsaturation index of the cellular membranes and accumulation of intracellular reactive oxygen species (ROS). This resulted in apoptosis, evidenced by reduced mitochondrial membrane potential and nuclear condensation and fragmentation. The most effective PUFA was stearidonic acid., Conclusions: The resultant cell death of both C. albicans and C. dubliniensis is due to apoptosis., General Significance: Due to the increase in drug resistance, alternative antifungal drugs are needed. A group of natural antifungal compounds is PUFAs. However, understanding their mechanisms of action is important for further use and development of these compounds as antifungal drugs. This paper provides insight into a possible mode of action of antifungal PUFAs., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Thrombotic thrombocytopenic purpura (TTP) is the prototypical microangiopathy characterized by disseminated microthromboses, hemolytic anemia, and ultimately organ dysfunction. A link with deficiency of the von Willebrand factor-cleaving protease (ADAMTS13) has been demonstrated, but additional genetic and/or environmental triggers are thought to be required to incite acute illness. Here we report that 4 days of ADAMTS13 functional inhibition is sufficient to induce TTP in the baboon (Papio ursinus), in the absence of inciting triggers because injections with an inhibitory monoclonal antibody (mAb) consistently (n = 6) induced severe thrombocytopenia (< 12 × 10(9)/L), microangiopathic hemolytic anemia, and a rapid rise in serum lactate dehydrogenase. Immunohistochemical staining revealed the characteristic disseminated platelet- and von Willebrand factor-rich thrombi in kidney, heart, brain, and spleen but not lungs. Prolonged inhibition (14 days, n = 1) caused myocardial ischemic damage and asplenia but not death. Control animals (n = 5) receiving equal doses of a noninhibitory anti-ADAMTS13 mAb remained unaffected. Our results provide evidence for a direct link between TTP and ADAMTS13 inhibition and for a mild disease onset. Furthermore, we present a reliable animal model of this disease as an opportunity for the development and validation of novel treatment strategies.

Two cases of violent, drug-intoxicated patients who presented considerable problems in management, and were resistant to standard sedative agents, are described. Effective and safe sedation, without the need for full anaesthesia and endotracheal intubation, was achieved using titrated doses of thiopentone. The merits of selecting this agent in these circumstances are discussed.

The processed leaves and stems of Aspalathus linearis contain a new diastereomeric pair of the flavanones, (S)- and (R)-eriodictyol-6-C-beta-D-glucopyranoside, which is also formed via the oxidative cyclization of the dihydrochalcone, aspalathin, under conditions which mimic the fermentation process.

Mushroom tyrosinase catalysed oxidation of three flavan-3-ols, viz. catechin, fisetinidol and mesquitol, was conducted to construct biphenyl bonds. Exposure of the flavan-3-ols to tyrosinase and subsequent trapping of the o-quinone intermediates resulted in the formation of novel flavan-3-ol derivatives, the structures of which were elucidated by mono- and two-dimensional 1H-NMR experiments. Application of the methodology resulted in the improved synthesis of the natural flavan-3-ol dimer, mesquitol-[5-->8]-catechin, previously isolated from Prosopis glandulosa.

Ovine keratoconjunctivitis was successfully reproduced in lambs under 1 year of age, in four separate transmission trials, by the use of mycoplasma isolates obtained from field outbreaks of ovine infectious keratoconjunctivitis. Mycoplasma isolates used in one of these trials, were identified as M. conjunctivae by means of immunofluorescence. Mycoplasma was isolated from approximately 87% of field cases examined. Branhamella ovis was isolated from 22% of field cases examined. No Chlamydia sp. or viruses were isolated from any of the outbreaks.

The aetiology and pathogenesis of respiratory syndromes in pigs are reviewed with emphasis on the role of environmental factors and diagnosis. Therapeutic and control measures are suggested and the cost of various regimens is dealt with in detail.