Intestine-Specific Mttp Deletion Decreases Mortality and Prevents Sepsis-Induced Intestinal Injury in a Murine Model of Pseudomonas aeruginosa Pneumonia

Background

The small intestine plays a crucial role in the pathophysiology of sepsis and has been referred to as the “motor” of the systemic inflammatory response. One proposed mechanism is that toxic gut-derived lipid factors, transported in mesenteric lymph, induce systemic injury and distant organ failure. However, the pathways involved are yet to be defined and the role of intestinal chylomicron assembly and secretion in transporting these lipid factors is unknown. Here we studied the outcome of sepsis in mice with conditional, intestine-specific deletion of microsomal triglyceride transfer protein (Mttp-IKO), which exhibit a block in chylomicron assembly together with lipid malabsorption.

Methodology/Principal Findings

Mttp-IKO mice and controls underwent intratracheal injection with either Pseudomonas aeruginosa or sterile saline. Mttp-IKO mice exhibited decreased seven-day mortality, with 0/20 (0%) dying compared to 5/17 (29%) control mice (p<0.05). This survival advantage in Mttp-IKO mice, however, was not associated with improvements in pulmonary bacterial clearance or neutrophil infiltration. Rather, Mttp-IKO mice exhibited protection against sepsis-associated decreases in villus length and intestinal proliferation and were also protected against increased intestinal apoptosis, both central features in control septic mice. Serum IL-6 levels, a major predictor of mortality in human and mouse models of sepsis, were elevated 8-fold in septic control mice but remained unaltered in septic Mttp-IKO mice. Serum high density lipoprotein (HDL) levels were reduced in septic control mice but were increased in septic Mttp-IKO mice. The decreased levels of HDL were associated with decreased hepatic expression of apolipoprotein A1 in septic control mice.

Conclusions/Significance

These studies suggest that strategies directed at blocking intestinal chylomicron secretion may attenuate the progression and improve the outcome of sepsis through effects mediated by metabolic and physiological adaptations in both intestinal and hepatic lipid flux.     

Induction of oscillatory ventilation pattern using dynamic modulation of heart rate through a pacemaker

For disease states characterized by oscillatory ventilation, an ideal dynamic therapy would apply a counteracting oscillation in ventilation. Modulating respiratory gas transport through the circulation might allow this. We explore the ability of repetitive alternations in heart rate, using a cardiac pacemaker, to elicit oscillations in respiratory variables and discuss the potential for therapeutic exploitation. By incorporating acute cardiac output manipulations into an integrated mathematical model, we observed that a rise in cardiac output should yield a gradual rise in end-tidal CO2 and, subsequently, ventilation. An alternating pattern of cardiac output might, therefore, create oscillations in CO2 and ventilation. We studied the effect of repeated alternations in heart rate of 30 beats/min with periodicity of 60 s, on cardiac output, respiratory gases, and ventilation in 22 subjects with implanted cardiac pacemakers and stable breathing patterns. End-tidal CO2 and ventilation developed consistent oscillations with a period of 60 s during the heart rate alternations, with mean peak-to-trough relative excursions of 8.4 +/- 5.0% (P < 0.0001) and 24.4 +/- 18.8% (P < 0.0001), respectively. Furthermore, we verified the mathematical prediction that the amplitude of these oscillations would depend on those in cardiac output (r = 0.59, P = 0.001). Repetitive alternations in heart rate can elicit reproducible oscillations in end-tidal CO2 and ventilation. The size of this effect depends on the magnitude of the cardiac output response. Harnessed and timed appropriately, this cardiorespiratory mechanism might be exploited to create an active dynamic responsive pacing algorithm to counteract spontaneous respiratory oscillations, such as those causing apneic breathing disorders.     

Community Health Seeking Behavior for Suspected Human and Animal Rabies Cases,Gomma District,Southwest Ethiopia

Background

Timely presentation to appropriate health service provider of sick animals/humans from zoonotic diseases like rabies is important for early case/outbreak detection and management. However, data on community’s health seeking practice for rabies in Ethiopia is limited. Therefore the objective of this study was to determine community’s health seeking behavior on rabies, Southwest Ethiopia.

Methods

A cross-sectional survey was conducted from January 16-February 14, 2015 to collect data from 808 respondents where the respondents were selected using multistage sampling technique. Data were collected using interviewer administered structured questionnaire by trained epidemiology graduate level students. Data were entered to Epidata version 3.1 and analyzed using SPSS version 20 for windows.

Result

Eight hundred three (99.4%) respondents participated in the study. Out of 28 respondents who reported their family members’ exposure to rabies, 8 of them replied that the exposed family members sought treatment from traditional healers. More than nine in ten respondents perceived that humans and domestic animals with rabies exposure should seek help of which 85% of them suggested modern health care facilities as the preferred management option for the sick humans and domestic animals. However, among those who reported sick domestic animals, near to 72% of them had either slaughtered for human consumption, sold immediately, visited traditional healer, given home care or did nothing for the sick domestic animals.

Conclusion

Majority of the respondents had favorable perception of seeking treatment from modern health care facilities for rabies. However, significant number of them had managed inappropriately for the sick domestic animals from rabies. Hence, raising awareness of the community about management of sick domestic animals from rabies and the need for reporting to both human and animal health service providers is needed.     

Genetic diversity in Pareuchaetes insulata and its implications for biological control of Chromolaena odorata

Chromolaena odorata (L.) King & H.E. Robins. (Asteraceae) threatens several economic and environmental activities in the eastern subtropical regions of South Africa and is a target for biological control. Three populations of Pareuchaetes insulata (Walker) (Lepidoptera: Erebidae) originating from Florida (USA), Cuba and Jamaica were released at 30 separate sites in KwaZulu-Natal, South Africa. Subsequent overlapping of two of these populations in the field and the likelihood of interbreeding posed a potential risk of establishment failure. To explore the genetic diversity of the different P. insulata populations and test for the existence of pre- and post-zygotic reproductive isolation between them, molecular analyses and cross-mating experiments were conducted. Mitochondrial DNA sequences revealed significant genetic similarity between them. Cross-mating trials between Floridian and Jamaican populations of P. insulata revealed no significant pre- and post-zygotic isolation, with no demonstrable differences in measured parameters between the ‘parental’ populations, the F₁ ‘hybrids’ and the backcrosses. These results suggest that P. insulata populations released in South Africa are part of the same ‘parental’ population. Genetic incompatibility is, therefore, relatively unlikely to be the cause of any failure in establishment of P. insulata at any of the release sites.     

Genetic diversity and differentiation of cultivated amochi (Arisaema schimperianum Schott) in Ethiopia as revealed by AFLP markers

Amochi (Arisaema schimperianum Schott) is an off-season crop plant in southern Ethiopia, grown during the dry season on residual moisture, for its edible tubers. It has gained importance as a "security crop" especially during the years of moisture stress and food shortage. Amochi is irritating in contact to the skin. Removal of this effect is an important question for breeding. As the first step, however we attempt to establish base line information of its breeding system and genetic variability using AFLPs. The extent of genetic differentiation among 11 populations (96 individuals) of amochi sampled along altitudinal gradients that varied from 1700 to 3200 m a.s.l. was investigated. The populations were classified in to three altitudinal groups: lowland (1700 to 2200 m a.s.l.), central-highland (2201 to 2600 m a.s.l.) and highland (2601 to 3200 m a.s.l.). Polymorphic loci (167) scored from four primer pair combinations, were used for principal component analysis (PCA), and analysis of molecular variance (AMOVA). Both PCA and unweighed pair group with arithmetic mean (UPGMA) clearly differentiated populations into their respective altitude groups, with large genetic distances. AMOVA analysis revealed 70.5%, 16.7% and 12.8% variability between altitude groups, between populations and within populations respectively. Average diversity indices within populations were also low. Since the largest proportion of variation is located between altitude groups, rather than within populations, we suggest future studies on the chemical composition, low irritation, and other desirable traits should consider populations from different altitude ranges.     

H-NS regulation of IraD and IraM antiadaptors for control of RpoS degradation

RpoS, the master sigma factor during stationary phase and under a variety of stress conditions, is regulated at multiple levels, including regulated degradation. Degradation is dependent upon ClpXP and the RssB adaptor protein. H-NS, a nucleoid-associated protein, affects the regulated degradation of RpoS; in the absence of H-NS, RpoS is stable. The mechanisms involved in this regulation were not known. We have found that H-NS inhibits the expression of iraD and iraM, the genes coding for two antiadaptor proteins that stabilize RpoS when overexpressed. The regulation by H-NS of iraM is independent from the previously demonstrated regulation by the PhoP/PhoQ two-component system. Moreover, differences in the behavior of several hns alleles are explained by a role for StpA, an H-NS-like protein, in the regulation of RpoS stability. This finding parallels recent observations for a role of StpA in regulation of RpoS stability in Salmonella.     

A controlled water-table depth system to study the influence of fine-scale differences in water regime for plant growth

A method was developed to maintain water-table depths at a constant level in outdoor mesocosms. The system included a water treatment reservoir, where tap water was microbially deoxygenated and denitrified; an adjustable-level control chamber that set desired water-table depths and plant growing mesocosms.The soil water status was evaluated by constant monitoring using tensiometers, pressure transducers and dipwells. The robustness of the system was tested by inducing sudden incidents of flooding and drainage. The system was able to revert to the original set water-table depths within 5 and 10 min, respectively. It also reliably sustained consistent water-table depths throughout the growing season without the need for maintenance.As an example, the method was used to grow plants at five set water-table depths: 50, 150, 250, 350, and 450 mm below ground surface. Two wet grassland species Festuca pratensis (meadow fescue), and Carex nigra (common sedge) were grown and dry biomass production recorded. Results showed differences in growth response between the two species to subjected water-table depths. In monoculture, F. pratensis production followed the order 50 = 150 = 350 > 250 = 450 mm (p < 0.001), while for C. nigra it was 150 = 250 > 50 = 350 = 450 mm (p < 0.001). In mixture, F. pratensis did not show a significant trend (p < 0.06), whereas C. nigra showed 50 = 150 > 250 > 350 = 450 mm (p < 0.001).The ease of the system to establish constant and/or dynamic water-table depths and its reliability outdoors renders it useful for a wide variety of studies involving plant growth.     

A Neuron-Based Screening Platform for Optimizing Genetically-Encoded Calcium Indicators

Fluorescent protein-based sensors for detecting neuronal activity have been developed largely based on non-neuronal screening systems. However, the dynamics of neuronal state variables (e.g., voltage, calcium, etc.) are typically very rapid compared to those of non-excitable cells. We developed an electrical stimulation and fluorescence imaging platform based on dissociated rat primary neuronal cultures. We describe its use in testing genetically-encoded calcium indicators (GECIs). Efficient neuronal GECI expression was achieved using lentiviruses containing a neuronal-selective gene promoter. Action potentials (APs) and thus neuronal calcium levels were quantitatively controlled by electrical field stimulation, and fluorescence images were recorded. Images were segmented to extract fluorescence signals corresponding to individual GECI-expressing neurons, which improved sensitivity over full-field measurements. We demonstrate the superiority of screening GECIs in neurons compared with solution measurements. Neuronal screening was useful for efficient identification of variants with both improved response kinetics and high signal amplitudes. This platform can be used to screen many types of sensors with cellular resolution under realistic conditions where neuronal state variables are in relevant ranges with respect to timing and amplitude.