Dynamic mathematical model development and validation of in vitro Mycobacterium smegmatis growth under nutrient- and pH-stress.

Mycobacterium tuberculosis can exist within a host for lengthy periods, tolerating even antibiotic challenge. This non-heritable, antibiotic tolerant "persister" state, is thought to underlie latent Tuberculosis (TB) infection and a deeper understanding thereof could inform treatment strategies. In addition to experimental studies, mathematical and computational modelling approaches are widely employed to study persistence from both an in vivo and in vitro perspective. However, specialized models (partial differential equations, agent-based, multiscale, etc.) rely on several difficult to determine parameters. In this study, a dynamic mathematical model was developed to predict the response of Mycobacterium smegmatis (a model organism for M. tuberculosis) grown in batch culture and subjected to a range of in vitro environmental stresses. Lag phase dynamics, pH variations and internal nitrogen storage were mechanistically modelled. Experimental results were used to train model parameters using global optimization, with extensive subsequent model validation to ensure extensibility to more complex modelling frameworks. This included an identifiability analysis which indicated that seven of the thirteen model parameters were uniquely identifiable. Non-identifiable parameters were critically evaluated. Model predictions compared to validation data (based on experimental results not used during training) were accurate with less than 16% maximum absolute percentage error, indicating that the model is accurate even when extrapolating to new experimental conditions. The bulk growth model can be extended to spatially heterogeneous simulations such as an agent-based model to simulate in vitro granuloma models or, eventually, in vivo conditions, where distributed environmental conditions are difficult to measure.

Characterization of smallholder pig production systems in the Western Cape, South Africa.

This study set out to characterize the farming and management practices of smallholder pig production systems in the Western Cape, South Africa. Data was gathered in three municipalities: Khayelitsha (27 farmers), Mamre (26 farmers) and Malmesbury (22 farmers) with the use of a structured questionnaire through both face-to-face and telephonic interviews. The study revealed that smallholder pig farmers in the Western Cape owned on average 37.4 ± 50.5 pigs. Majority of farmers (61%) made use of tattoo markings to identify their pigs, 6% made use of ear tags and 4% clipped the ears of their pigs. More than 25% of farmers did not use any method of identification to mark their pigs. More than half of the respondents (51%) do not have formal recordkeeping systems in place. Livestock theft was found to be a major problem amongst farmers. No differences were found amongst the three study areas and the type of housing system used (P > 0.05). The most popular materials pens were constructed of across the three areas were wood (40%), corrugated iron (16%) or a combination of the two (32%); other materials used were cement blocks (5%), polystyrene (1%) or plastic (1%). Pig farmers kept 43% Large White, 43 % crossbred and 14% Landrace across the three districts. Commercial feed was used by 61% of Mamre farmers whereas Khayelitsha farmers fed an equal amount of by-products (44%) and mixed (44%) feed to their pigs, whereas Malmesbury fed 44% mixed, 33% commercial and 24% by-products obtained from factories to their pigs. Piglet mortality due to the sow lying on her piglets was experienced by 47% of farmers interviewed, whilst 29% had experienced piglet mortalities due to the cold. Perturbingly, 76% of farmers did not send dead pigs for a post-mortem, stating that they could not rely on veterinarians or afford their services. Smallholder pig farmers in Malmesbury (68%) and Khayelitsha (70%) sold their pigs to the informal market, indicating that keeping pigs provided a source of revenue. Mamre marketed 88% of their pigs as weaners, Khayelitsha marketed 73% of their pigs as growers, whereas Malmesbury kept 32% of their pigs as breeding stock. Khayelitsha and Malmesbury marketed their growers at 63.3 ± 41.2 kg and 51.5 ± 30.2 kg, respectively, to the informal market. The study highlights the farming practices of smallholder pig farmers in three districts in the Western Cape. There a few differences in the practices of these farmers, but overall, there are similar limitations that hinder them from improved production, such as limited recordkeeping, theft, lack of infrastructure and finances for post-mortem examination. Smallholder pig farmers in the Western Cape are still mainly selling to the informal market. This informal market is not fully characterized yet in the Western Cape, and more studies are needed to further determine and investigate the benefits of only selling to the informal market.

Immobilization quality and cardiopulmonary effects of etorphine alone compared with etorphine-azaperone in blesbok (Damaliscus pygargus phillipsi).

OBJECTIVE: To evaluate the immobilization quality and cardiopulmonary effects of etorphine alone compared with etorphine-azaperone in blesbok (Damaliscus pygargus phillipsi). STUDY DESIGN: Blinded, randomized, crossover design. ANIMALS: A total of 12 boma-habituated female blesbok weighing [mean ± standard deviation (SD)] 57.5 ± 2.5 kg. METHODS: Each animal was administered etorphine (0.09 mg kg-1) or etorphine-azaperone (0.09 mg kg-1; 0.35 mg kg-1) intramuscularly with 1-week intertreatment washout period. Time to first sign of altered state of consciousness and immobilization time were recorded. Physiological variables were recorded, arterial blood samples were taken during a 40-minute immobilization period, and naltrexone (mean ± SD: 1.83 ± 0.06 mg kg-1) was intravenously administered. Recovery times were documented, and induction, immobilization and recovery were subjectively scored. Statistical analyses were performed; p < 0.05 was significant. RESULTS: No difference was observed in time to first sign, immobilization time and recovery times between treatments. Time to head up was longer with etorphine-azaperone (0.5 ± 0.2 versus 0.4 ± 0.2 minutes; p = 0.015). Etorphine caused higher arterial blood pressures (mean: 131 ± 17 versus 110 ± 11 mmHg, p < 0.0001), pH, rectal temperature and arterial oxygen partial pressure (59.2 ± 7.7 versus 42.2 ± 9.8 mmHg), but lower heart (p = 0.002) and respiratory rates (p = 0.01). Etorphine-azaperone combination led to greater impairment of ventilatory function, with higher end-tidal carbon dioxide (p < 0.0001) and arterial partial pressure of carbon dioxide (58.0 ± 4.5 versus 48.1 ± 5.1 mmHg). Immobilization quality was greater with etorphine-azaperone than with etorphine alone (median scores: 4 versus 3; p < 0.0001). CONCLUSIONS AND CLINICAL RELEVANCE: Both treatments provided satisfactory immobilization of blesbok; however, in addition to a deeper level of immobilization, etorphine-azaperone caused greater ventilatory impairment. Oxygen supplementation is recommended with both treatments.

Gender differences in tuberculosis treatment outcomes: a post hoc analysis of the REMoxTB study.

BACKGROUND: In the REMoxTB study of 4-month treatment-shortening regimens containing moxifloxacin compared to the standard 6-month regimen for tuberculosis, the proportion of unfavourable outcomes for women was similar in all study arms, but men had more frequent unfavourable outcomes (bacteriologically or clinically defined failure or relapse within 18 months after randomisation) on the shortened moxifloxacin-containing regimens. The reason for this gender disparity in treatment outcome is poorly understood. METHODS: The gender differences in baseline variables were calculated, as was time to smear and culture conversion and Kaplan-Meier plots were constructed. In post hoc exploratory analyses, multivariable logistic regression modelling and an observed case analysis were used to explore factors associated with both gender and unfavourable treatment outcome. RESULTS: The per-protocol population included 472/1548 (30%) women. Women were younger and had lower rates of cavitation, smoking and weight (all p < 0.05) and higher prevalence of HIV (10% vs 6%, p = 0.001). They received higher doses (mg/kg) than men of rifampicin, isoniazid, pyrazinamide and moxifloxacin (p ≤ 0.005). There was no difference in baseline smear grading or mycobacterial growth indicator tube (MGIT) time to positivity. Women converted to negative cultures more quickly than men on Lowenstein-Jensen (HR 1.14, p = 0.008) and MGIT media (HR 1.19, p < 0.001). In men, the presence of cavitation, positive HIV status, higher age, lower BMI and 'ever smoked' were independently associated with unfavourable treatment outcome. In women, only 'ever smoked' was independently associated with unfavourable treatment outcome. Only for cavitation was there a gender difference in treatment outcomes by regimen; their outcome in the 4-month arms was significantly poorer compared to the 6-month treatment arm (p < 0.001). Women, with or without cavities, and men without cavities had a similar outcome on all treatment arms (p = 0.218, 0.224 and 0.689 respectively). For all other covariate subgroups, there were no differences in treatment effects for men or women. CONCLUSIONS: Gender differences in TB treatment responses for the shorter regimens in the REMoxTB study may be explained by poor outcomes in men with cavitation on the moxifloxacin-containing regimens. We observed that women with cavities, or without, on the 4-month moxifloxacin regimens had similar outcomes to all patients on the standard 6-month treatment. The biological reasons for this difference are poorly understood and require further exploration.

Muscle fiber type and metabolic profiles of four muscles from the African black ostrich.

Muscle fiber type, fiber cross-sectional area (CSA), enzyme activities (citrate synthase (CS), 3-hydroxyacetyl Co A dehydrogenase (3HAD), lactate dehydrogenase (LDH) and phosphofructokinase (PFK)) and glycogen content were analyzed in the M. iliotibialis cranialis (ITC), M. iliotibialis lateralis, M. gastrocnemius (G) and M. fibularis longus (FL) muscles from 24 ostriches. Type I and II fiber proportions were similar across the 4 muscles, but the ITC had overall the smallest fibers. CS activity was the highest in the ITC, but similar between the remainder of the muscles. 3HAD activities were very low in all muscles, ranging between 1.9 and 2.7 µmol/min/g protein, indicating poor ß-oxidation. The ITC also had the lowest PFK activity. Glycogen content averaged ∼85 mmol/kg dry weight across the muscles with large intramuscular variations. The 4 ostrich muscles present with low fat oxidation capacity and low glycogen content, which could have significant implications on meat quality attributes.

Knowledge of stakeholders in the game meat industry and its effect on compliance with food safety standards.

The game meat industry is continuing to grow in South Africa. Several stakeholders are involved in the game meat supply chain and a high level of knowledge is necessary to ensure compliance with legislation and standards. It was therefore necessary to determine the level of knowledge of the stakeholders since this has not been determined before. Information regarding the extent of stakeholders' knowledge and the possible impact on compliance to standards was obtained through a desk-top study and an analysis of questionnaire responses from industry, consumers and relevant authorities. Results have shown that consumers have a specific expectation regarding the safe production of game meat. Limitations in the knowledge of the stakeholders have been identified. Understanding these limitations can assist policy-makers, law enforcers and the game meat industry in developing strategies to alleviate the problem. The result of this study may assist in providing consumers with game meat that is safe for human consumption.

A partially randomised trial of pretomanid, moxifloxacin and pyrazinamide for pulmonary TB.

BACKGROUND: Treatment for TB is lengthy and toxic, and new regimens are needed.METHODS: Participants with pulmonary drug-susceptible TB (DS-TB) were randomised to receive: 200 mg pretomanid (Pa, PMD) daily, 400 mg moxifloxacin (M) and 1500 mg pyrazinamide (Z) for 6 months (6Pa200MZ) or 4 months (4Pa200MZ); 100 mg pretomanid daily for 4 months in the same combination (4Pa100MZ); or standard DS-TB treatment for 6 months. The primary outcome was treatment failure or relapse at 12 months post-randomisation. The non-inferiority margin for between-group differences was 12.0%. Recruitment was paused following three deaths and not resumed.RESULTS: Respectively 4/47 (8.5%), 11/57 (19.3%), 14/52 (26.9%) and 1/53 (1.9%) DS-TB outcomes were unfavourable in patients on 6Pa200MZ, 4Pa200MZ, 4Pa100MZ and controls. There was a 6.6% (95% CI -2.2% to 15.4%) difference per protocol and 9.9% (95%CI -4.1% to 23.9%) modified intention-to-treat difference in unfavourable responses between the control and 6Pa200MZ arms. Grade 3+ adverse events affected 68/203 (33.5%) receiving experimental regimens, and 19/68 (27.9%) on control. Ten of 203 (4.9%) participants on experimental arms and 2/68 (2.9%) controls died.CONCLUSION: PaMZ regimens did not achieve non-inferiority in this under-powered trial. An ongoing evaluation of PMD remains a priority.

Feather and skin development of ostriches Struthio camelus.

Information on feather and skin growth is important for the development of mathematical optimisation nutritional models for ostriches. Ostriches (n = 65) were subjected to a four-stage formulated growth diet programme (pre-starter, starter, grower and finisher), with declining protein and energy content. Nine birds were weighed, stunned, exsanguinated, defeathered, skinned and eviscerated at 1, 54, 84, 104, 115, 132 and 287 days of age. Feathers from four pre-selected locations on the body were harvested and weighed. The wet skin weight, wet unstretched skin size and wet unstretched crown size were measured at each slaughter stage. The live weight, feather and skin yields of the birds increased with age at slaughter, as did feather shaft diameter. Prediction models were developed to estimate the yield of the skin in terms of live weight and of empty body protein weight to aid in diet formulation. The allometry of feather growth was determined from total feather weight, as the maturation rates of the feathers differ from that of the ostrich body. Results from this study will aid in setting up a mathematical optimisation nutritional model for ostriches.

The effect of polysaccharide-degrading wine yeast transformants on the efficiency of wine processing and wine flavour.

Commercial polysaccharase preparations are applied to winemaking to improve wine processing and quality. Expression of polysaccharase-encoding genes in Saccharomyces cerevisiae allows for the recombinant strains to degrade polysaccharides that traditional commercial yeast strains cannot. In this study, we constructed recombinant wine yeast strains that were able to degrade the problem-causing grape polysaccharides, glucan and xylan, by separately integrating the Trichoderma reesei XYN2 xylanase gene construct and the Butyrivibrio fibrisolvens END1 glucanase gene cassette into the genome of the commercial wine yeast strain S. cerevisiae VIN13. These genes were also combined in S. cerevisiae VIN13 under the control of different promoters. The strains that were constructed were compared under winemaking conditions with each other and with a recombinant wine yeast strain expressing the endo-beta-1,4-glucanase gene cassette (END1) from B. fibrisolvens and the endo-beta-1,4-xylanase gene cassette (XYN4) from Aspergillus niger, a recombinant strain expressing the pectate lyase gene cassette (PEL5) from Erwinia chrysanthemi and the polygalacturonase-encoding gene cassette (PEH1) from Erwinia carotovora. Wine was made with the recombinant strains using different grape cultivars. Fermentations with the recombinant VIN13 strains resulted in significant increases in free-flow wine when Ruby Cabernet must was fermented. After 6 months of bottle ageing significant differences in colour intensity and colour stability could be detected in Pinot Noir and Ruby Cabernet wines fermented with different recombinant strains. After this period the volatile composition of Muscat d'Alexandria, Ruby Cabernet and Pinot Noir wines fermented with different recombinant strains also showed significant differences. The Pinot Noir wines were also sensorial evaluated and the tasting panel preferred the wines fermented with the recombinant strains.

Characteristics of impala (Aepyceros melampus) skeletal muscles.

The aim of this study was to determine citrate synthase (CS), phosphofructokinase (PFK) activities and myosin heavy chain (MHC) isoform contents in four muscle groups (semimembranosus (S), deltoideus (D), longissimus lumborum (LL), and psoas major (PM)) of impala (n=6). All four muscle groups expressed predominantly MHC IIa (means of 55±22-93±12%). MHC IIx was only expressed in D. In D, positive correlations were found between MHC I and age (r=0.93; P<0.05) and the weight of the animals (r=0.94; P<0.01). PFK (means of 175±179-374±181), CS (means of 100±23-126±38 µmol/min/gdw) and MHC content indicated that energy provision in the impala is produced to a large extent via oxidative pathways and fibre types vary with animal characteristics.

Suspected lead poisoning in two captive cheetahs (Acinonyx jubatus jubatus) in South Africa, in 2008 and 2013.

Whilst lead poisoning in raptors, scavenging birds and waterfowl is well studied and common knowledge, there is surprisingly little literature detailing the risk to mammalian scavengers and captive carnivores fed hunted meat. This case report describes the death of two captive cheetahs (Acinonyx jubatus jubatus) following acute onset of nervous symptoms. Clinical signs included hyper-excitability, seizures, arched back, tail held abnormally high and hyper-salivation. Necropsy findings included bullets or a bullet in their stomachs. Kidney and liver lead levels from one cheetah (15.6 ppm and 17 ppm respectively) were consistent with a diagnosis of lead poisoning; liver from the second cheetah was not available for testing. Both animals were routinely fed hunted antelope or game birds. This is the first report of oral lead poisoning in captive large carnivores, although these are unlikely to be the first cases. Without awareness of the risks of feeding hunted game, lead exposure will continue to be an underdiagnosed reality in the rehabilitation of endangered carnivores.

Geotrichum candidum P-5 produces an intracellular serine protease resembling chymotrypsin.

A wide range of intra- and extracellular microbial proteases has been studied and characterized. These enzymes are mostly extracellular and in some cases they may resemble 'classical' serine proteases. As part of a programme in which the lipase and protease activities of the fungus Geotrichum candidum are being studied, an intracellular protease with an apparent chymotrypsin-like specificity was detected. The serine protease was isolated from biomass using ion-exchange and exclusion chromatography. Kinetic characterization was done using a series of synthetic substrates and inhibitors. Aprotinin-sepharose affinity chromatography was used to isolate a fraction for molecular size determination on SDS-PAGE. The purified protease, which could hydrolyse haemoglobin as protein substrate, was obtained with a 30-fold purification and a yield of 44%, but it was very unstable and rapidly lost activity. The enzyme which bound to the affinity column had a single subunit mass of 278 kDa. Kinetic analysis showed a similarity with trypsin and chymotrypsin, but tending more towards chymotrypsin in that a bulky aromatic group, e.g. phenylalanine in the P1 position, was preferred. The optimum pH was in the region of 7-8.25. Inhibition patterns indicated that the enzyme was a serine protease with no metal dependence, although it was stabilized by magnesium ions. The enzyme seems to share some properties with other intra- and extracellular microbial serine proteases. The exact function of the enzymatic activity is still unclear, but it is suggested that it may be involved with intracellular protein turnover.

Medicinal bulbous plants of South Africa and their traditional relevance in the control of infectious diseases.

Natural products are becoming more important in modern-day society as man is moving away from synthetic products, which can be detrimental to the environment and human health. Scientific research on the healing properties and bioactivity of natural compounds, especially of plant origin, has been extensive particularly in the Western world. However, a rich heritage of floral biodiversity is found in developing countries. South Africa, a country with a strong history of traditional healing, hosts a variety of around 30000 plant species. Indigenous bulbous plants of importance to South African traditional healers mainly belong to the Amaryllidaceae and Hyacinthaceae families. A number of these plants have particular uses as disinfectants and anti-inflammatory agents, although there is still a lack of scientific research regarding their unique pharmacological compounds.

The hygiene practices of three systems of game meat production in South Africa in terms of animal class and health compliance.

Three game meat production systems used on game ranches in South Africa are reported on. System one is applied in the game export market and conforms to the hygiene requirements of the European Union (EU). System two and three entail game meat available on the local market not subjected to any regulation. System 2 however, implemented basic meat hygiene values. Measurements of pH, temperature, Aerobic Plate Count (APC), E. coli, Salmonella and S. aureus were subjected to a 3×2 factorial analysis of variance with factors that involve 3 system compliances in 2 classes of game animals in a completely randomised design. The measured bacteriological and quality differences between the three systems do not justify EU standards application on the local market but results indicated a significant compliance×class interaction.

Two sampling techniques for game meat.

A study was conducted to compare the excision sampling technique used by the export market and the sampling technique preferred by European countries, namely the biotrace cattle and swine test. The measuring unit for the excision sampling was grams (g) and square centimetres (cm2) for the swabbing technique. The two techniques were compared after a pilot test was conducted on spiked approved beef carcasses (n = 12) that statistically proved the two measuring units correlated. The two sampling techniques were conducted on the same game carcasses (n = 13) and analyses performed for aerobic plate count (APC), Escherichia coli and Staphylococcus aureus, for both techniques. A more representative result was obtained by swabbing and no damage was caused to the carcass. Conversely, the excision technique yielded fewer organisms and caused minor damage to the carcass. The recovery ratio from the sampling technique improved 5.4 times for APC, 108.0 times for E. coli and 3.4 times for S. aureus over the results obtained from the excision technique. It was concluded that the sampling methods of excision and swabbing can be used to obtain bacterial profiles from both export and local carcasses and could be used to indicate whether game carcasses intended for the local market are possibly on par with game carcasses intended for the export market and therefore safe for human consumption.

Investigating the contributing factors to postmortem pH changes in springbok, eland, red hartebeest and kudu edible offal.

The objective of the study was to assess pH measurements between offal organs of different species and the association between pH taken 4 h post-slaughter and different predictor variables in the liver and lungs. A linear regression analysis was conducted on selected variables to identify the main predictors and their interactions affecting the pH of meat 4 h post-slaughter. In an increasing order of magnitude during winter, the pH achieved at 16 h - 36 h post-slaughter in springbok heart, liver, spleen, kidney and lungs was significantly (p < 0.05) higher than pH 6.0. The pH attained in springbok carcasses was (p < 0.05) below 6.0, whilst no significant differences were observed from the regulatory reference (pH 6.0) in the heart. There was a positive association between the pH of game meat 4 h post-slaughter and liver congestion. The pH of game meat 4 h post-slaughter increased by 0.11 units (p < 0.05) per millilitre increase in liver congestion and decreased by 0.04 units (p< 0.05) per minute increase in the shooting-to-bleeding interval, irrespective of the species. The lack of a statistically significant association between some selected variables and pH changes in this study suggested that either the factors may have a small effect which is only detectable with large data-sets and/or the effect may be modified by other unidentified factors. As some of the offal organs had final pH readings above 6.0, alternative measures are required to inactivate certain endogenous pathogens in edible wild game offal sourced from endemic areas.

Wildlife-associated zoonotic diseases in some southern African countries in relation to game meat safety: a review.

With on-going changes in land use practices from conventional livestock farming to commercial, wildlife-based activities, the interface or interaction between livestock and wildlife is increasing. As part of the wildlife-based activities of ecotourism, breeding and hunting, game farmers are also exploring the utilisation of meat from hunted or harvested game. The expanding interface or increased interaction between livestock and wildlife increases the risk of disease incidence and the emergence of new diseases or the re-emergence of previously diagnosed diseases. The risk is not only related to domestic and wild animal health, but also to the occupational hazards that it poses to animal handlers and the consumers of game meat. This review endeavours to highlight the role that game plays in the spreading of zoonotic diseases to other animals and humans. Examples of zoonotic diseases that have occurred in wild animals in the past, their relevance and risk have been summarised and should function as a quick reference guide for wildlife veterinarians, ecologists, farmers, hunters, slaughter staff, processors and public health professionals.

Zoonoses: a potential obstacle to the growing wildlife industry of Namibia.

Zoonoses, which account for approximately 75% of emerging human infectious diseases worldwide, pose a re-emerging threat to public health. With an ever-increasing interrelationship between humans, livestock and wildlife species, the threat to human health will rise to unprecedented levels. Wildlife species contribute to the majority of emerging diseases; therefore, there is an urgent need to define control systems of zoonoses of wildlife origin but very little information exists. In this review, we examine prevalent zoonotic infections reported in Namibia between 1990 and 2009 and assess their potential impact on the growing wildlife industry. A wide spectrum of zoonotic diseases was confirmed in both livestock and wildlife species, with rabies and anthrax cases being over-represented and also showing the widest species distribution. Whilst vaccination and ante-mortem inspection against these diseases may curb infected livestock species from entering the human food chain, such practices are difficult to implement in free-ranging wildlife species. In this context, there is a need to improve existing control measures and/or develop novel and better interventional strategies to reduce the threat of this re-emerging global problem. This review provides the basis for initiating a multidisciplinary evidence-based approach to control zoonoses in countries with thriving wildlife and game farming.

Efficacy and immunogenicity of two or three dose rotavirus-vaccine regimen in South African children over two consecutive rotavirus-seasons: a randomized, double-blind, placebo-controlled trial.

BACKGROUND: Human rotavirus vaccine (HRV; i.e., Rotarix) reduced the incidence of severe rotavirus gastroenteritis (RVGE) by 77% (95% Confidence interval: 56-88%) during the first year of life in South Africa. Persistence of HRV-derived protection against RVGE during subsequent rotavirus seasons, although evident in industrialized settings, remains to be established in African settings. This study reports on the efficacy of HRV against severe RVGE over two consecutive rotavirus seasons in South African children. METHODS: A prospective, double-blind, placebo controlled multi-centered trial in South Africa and Malawi randomly assigned infants in a 1:1:1 ratio to receive either two (10 and 14 weeks; HRV_2D) or three (6, 10 and 14 weeks; HRV_3D) doses of HRV or placebo. The primary analysis involved pooling of HRV_2D and HRV_3D arms. Episodes of gastroenteritis caused by wild-type rotavirus were identified through active follow-up surveillance and graded by the Vesikari scale. RESULTS: 1339 infants (447 in the HRV_2D group, 447 in the HRV_3D group and 445 in the placebo group) were enrolled in Year 2 of the study, including 1035 (77.3%) who were followed up over two consecutive rotavirus seasons (i.e., Cohort 2 subjects). Rotarix was associated with ongoing protection against severe RVGE, preventing 2.5 episodes per 100 vaccinated children over two consecutive rotavirus seasons; vaccine efficacy: 59% (95% Confidence interval: 1-83%). An exploratory analysis indicated better immunogenicity (among Cohort 1 subjects) and a higher point-efficacy estimate over two seasons in the HRV_3D compared to HRV_2D arms of the study in Cohort 2 subjects. CONCLUSION: Rotarix is associated with significant reductions in severe gastroenteritis episodes through 2 years of life among South African children. Further research is needed to determine the optimal dosing schedule of Rotarix in providing long-term protection against rotavirus illness in African children.

Roflumilast, a phosphodiesterase 4 inhibitor, reduces airway hyperresponsiveness after allergen challenge.

BACKGROUND: Roflumilast, an oral, once-daily phosphodiesterase 4 inhibitor, is currently in clinical development for the treatment of asthma. OBJECTIVES: This pilot study examined the effect of roflumilast on allergen-induced airway hyperresponsiveness (AHR) to histamine challenge and asthmatic response to allergen challenge. METHODS: In a randomized, double-blind, 2-period, crossover trial, 13 patients with mild allergic asthma [mean forced expiratory volume in 1 s (FEV(1)) % predicted = 86%] received a single dose of oral roflumilast 1,000 microg or placebo. Patients were administered roflumilast 60 min before allergen challenge, and asthmatic responses were assessed via change in FEV(1)