Rabbit model of uncontrolled hemorrhagic shock and hypotensive resuscitation

Clinically relevant animal models capable of simulating traumatic hemorrhagic shock are needed. We developed a hemorrhagic shock model with male New Zealand rabbits (2200-2800 g, 60-70 days old) that simulates the pre-hospital and acute care of a penetrating trauma victim in an urban scenario using current resuscitation strategies. A laparotomy was performed to reproduce tissue trauma and an aortic injury was created using a standardized single puncture to the left side of the infrarenal aorta to induce hemorrhagic shock similar to a penetrating mechanism. A 15-min interval was used to simulate the arrival of pre-hospital care. Fluid resuscitation was then applied using two regimens: normotensive resuscitation to achieve baseline mean arterial blood pressure (MAP, 10 animals) and hypotensive resuscitation at 60% of baseline MAP (10 animals). Another 10 animals were sham operated. The total time of the experiment was 85 min, reproducing scene, transport and emergency room times. Intra-abdominal blood loss was significantly greater in animals that underwent normotensive resuscitation compared to hypotensive resuscitation (17.1 ± 2.0 vs 8.0 ± 1.5 mL/kg). Antithrombin levels decreased significantly in normotensive resuscitated animals compared to baseline (102 ± 2.0 vs 59 ± 4.1%), sham (95 ± 2.8 vs 59 ± 4.1%), and hypotensive resuscitated animals (98 ± 7.8 vs 59 ± 4.1%). Evidence of re-bleeding was also noted in the normotensive resuscitation group. A hypotensive resuscitation regimen resulted in decreased blood loss in a clinically relevant small animal model capable of reproducing hemorrhagic shock caused by a penetrating mechanism.

Induction of Zenk protein expression within the nucleus taeniae of the amygdala of pigeons following tone and shock stimulation

In this study, we evaluated the expression of the Zenk protein within the nucleus taeniae of the pigeon’s amygdala (TnA) after training in a classical aversive conditioning, in order to improve our understanding of its functional role in birds. Thirty-two 18-month-old adult male pigeons (Columba livia), weighing on average 350 g, were trained under different conditions: with tone-shock associations (experimental group; EG); with shock-alone presentations (shock group; SG); with tone-alone presentations (tone group; TG); with exposure to the training chamber without stimulation (context group; CG), and with daily handling (naive group; NG). The number of immunoreactive nuclei was counted in the whole TnA region and is reported as density of Zenk-positive nuclei. This density of Zenk-positive cells in the TnA was significantly greater for the EG, SG and TG than for the CG and NG (P < 0.05). The data indicate an expression of Zenk in the TnA that was driven by experience, supporting the role of this brain area as a critical element for neural processing of aversive stimuli as well as meaningful novel stimuli.

Effect of ATP and 2-oxoglutarate on the in vitro interaction between the NifA GAF domain and the GlnB protein of Azospirillum brasilense

Azospirillum brasilense is a diazotroph that associates with important agricultural crops and thus has potential to be a nitrogen biofertilizer. The A. brasilense transcription regulator NifA, which seems to be constitutively expressed, activates the transcription of nitrogen fixation genes. It has been suggested that the nitrogen status-signaling protein GlnB regulates NifA activity by direct interaction with the NifA N-terminal GAF domain, preventing the inhibitory effect of this domain under conditions of nitrogen fixation. In the present study, we show that an N-terminal truncated form of NifA no longer required GlnB for activity and lost regulation by ammonium. On the other hand, in trans co-expression of the N-terminal GAF domain inhibited the N-truncated protein in response to fixed nitrogen levels. We also used pull-down assays to show in vitro interaction between the purified N-terminal GAF domain of NifA and the GlnB protein. The results showed that A. brasilense GlnB interacts directly with the NifA N-terminal domain and this interaction is dependent on the presence of ATP and 2-oxoglutarate.

Myosin light chain kinase is necessary for post-shock mesenteric lymph drainage enhancement of vascular reactivity and calcium sensitivity in hemorrhagic-shocked rats

Vascular hyporeactivity is an important factor in irreversible shock, and post-shock mesenteric lymph (PSML) blockade improves vascular reactivity after hemorrhagic shock. This study explored the possible involvement of myosin light chain kinase (MLCK) in PSML-mediated vascular hyporeactivity and calcium desensitization. Rats were divided into sham (n=12), shock (n=18), and shock+drainage (n=18) groups. A hemorrhagic shock model (40±2 mmHg, 3 h) was established in the shock and shock+drainage groups. PSML drainage was performed from 1 to 3 h from start of hypotension in shock+drainage rats. Levels of phospho-MLCK (p-MLCK) were determined in superior mesenteric artery (SMA) tissue, and the vascular reactivity to norepinephrine (NE) and sensitivity to Ca2+ were observed in SMA rings in an isolated organ perfusion system. p-MLCK was significantly decreased in the shock group compared with the sham group, but increased in the shock+drainage group compared with the shock group. Substance P (1 nM), an agonist of MLCK, significantly elevated the decreased contractile response of SMA rings to both NE and Ca2+ at various concentrations. Maximum contractility (Emax) in the shock group increased with NE (from 0.179±0.038 to 0.440±0.177 g/mg, P<0.05) and Ca2+ (from 0.515±0.043 to 0.646±0.096 g/mg, P<0.05). ML-7 (0.1 nM), an inhibitor of MLCK, reduced the increased vascular response to NE and Ca2+ at various concentrations in the shock+drainage group (from 0.744±0.187 to 0.570±0.143 g/mg in Emax for NE and from 0.729±0.037 to 0.645±0.056 g/mg in Emax for Ca2+, P<0.05). We conclude that MLCK is an important contributor to PSML drainage, enhancing vascular reactivity and calcium sensitivity in rats with hemorrhagic shock.

The N-terminal 33 amino acid domain of Siva-1 is sufficient for nuclear localization

Siva-1 induces apoptosis in multiple pathological processes and plays an important role in the suppression of tumor metastasis, protein degradation, and other functions. Although many studies have demonstrated that Siva-1 functions in the cytoplasm, a few have found that Siva-1 can relocate to the nucleus. In this study, we found that the first 33 amino acid residues of Siva-1 are required for its nuclear localization. Further study demonstrated that the green fluorescent protein can be imported into the nucleus after fusion with these 33 amino acid residues. Other Siva-1 regions and domains showed less effect on Siva-1 nuclear localization. By site-mutagenesis of all of these 33 amino acid residues, we found that mutants of the first 1-18 amino acids affected Siva-1 nuclear compartmentalization but could not complete this localization independently. In summary, we demonstrated that the N-terminal 33 amino acid residues were sufficient for Siva-1 nuclear localization, but the mechanism of this translocation needs additional investigation.

Mesenteric lymph reperfusion exacerbates spleen injury caused by superior mesenteric artery occlusion shock

The intestinal lymph pathway plays an important role in the pathogenesis of organ injury following superior mesenteric artery occlusion (SMAO) shock. We hypothesized that mesenteric lymph reperfusion (MLR) is a major cause of spleen injury after SMAO shock. To test this hypothesis, SMAO shock was induced in Wistar rats by clamping the superior mesenteric artery (SMA) for 1 h, followed by reperfusion for 2 h. Similarly, MLR was performed by clamping the mesenteric lymph duct (MLD) for 1 h, followed by reperfusion for 2 h. In the MLR+SMAO group rats, both the SMA and MLD were clamped and then released for reperfusion for 2 h. SMAO shock alone elicited: 1) splenic structure injury, 2) increased levels of malondialdehyde, nitric oxide (NO), intercellular adhesion molecule-1, endotoxin, lipopolysaccharide receptor (CD14), lipopolysaccharide-binding protein, and tumor necrosis factor-α, 3) enhanced activities of NO synthase and myeloperoxidase, and 4) decreased activities of superoxide dismutase and ATPase. MLR following SMAO shock further aggravated these deleterious effects. We conclude that MLR exacerbates spleen injury caused by SMAO shock, which itself is associated with oxidative stress, excessive release of NO, recruitment of polymorphonuclear neutrophils, endotoxin translocation, and enhanced inflammatory responses.

Hydrogen sulfide in posthemorrhagic shock mesenteric lymph drainage alleviates kidney injury in rats

Posthemorrhagic shock mesenteric lymph (PHSML) is a key factor in multiple organ injury following hemorrhagic shock. We investigated the role of hydrogen sulfide (H2S) in PHSML drainage in alleviating acute kidney injury (AKI) by administering D,L-propargylglycine (PPG) and sodium hydrosulfide hydrate (NaHS) to 12 specific pathogen-free male Wistar rats with PHSML drainage. A hemorrhagic shock model was established in 4 experimental groups: shock, shock+drainage, shock+drainage+PPG (45 mg/kg, 0.5 h prehemorrhage), and shock+drainage+NaHS (28 µmol/kg, 0.5 h prehemorrhage). Fluid resuscitation was performed after 1 h of hypotension, and PHMSL was drained in the last three groups for 3 h after resuscitation. Renal function and histomorphology were assessed along with levels of H2S, cystathionine-γ-lyase (CSE), Toll-like receptor 4 (TLR4), interleukin (IL)-10, IL-12, and tumor necrosis factor (TNF)-α in renal tissue. Hemorrhagic shock induced AKI with increased urea and creatinine levels in plasma and higher H2S, CSE, TLR4, IL-10, IL-12, and TNF-α levels in renal tissue. PHSML drainage significantly reduced urea, creatinine, H2S, CSE, and TNF-α but not TLR4, IL-10, or IL-12. PPG decreased creatinine, H2S, IL-10, and TNF-α levels, but this effect was reversed by NaHS administration. In conclusion, PHSML drainage alleviated AKI following hemorrhagic shock by preventing increases in H2S and H2S-mediated inflammation.

Weldability of powder bed fusion fabricated stainless steel 316L sheets to cold rolled sheet metal

Weldability of powder bed fusion (PBF) fabricated components has come to discussion in past two years due to resent developments in the PBF technology and limited size of the machines used in the fabrication process. This study concentrated on effects of energy input of welding on mechanical properties and microstructural features of welds between PBF fabricated stainless steel 316L sheets and cold rolled sheet metal of same composition by the means of destructive testing and microscopic analysis. Optical fiber diameter, laser power and welding speed were varied during the experiments that were executed following one variable at a time (OVAT) method. One of the problems of welded PBF fabricated components has been lower elongations at break comparing to conventionally manufactured components. Decreasing energy input of the laser keyhole welding decreased elongations at break of the welded specimens. Ultimate tensile strengths were not affected significantly by the energy input of the welding, but fracturing of the specimens welded using high energy input occurred from the weld metal. Fracturing of the lower energy input welds occurred from the PBF fabricated base metal. Energy input was found to be critical factor for mechanical properties of the welds. Multioriented grain growth and formation of neck at fusion zone boundary on the cold rolled side of the weld was detected and suspected to be result from weld pool flows caused by differences in molten weld pool behaviour between the PBF fabricated and cold rolled sides of the welds.

Sensory profile of 'Douradão' peaches cold stored under controlled atmosphere

The sensory quality of 'Douradão' peaches cold stored in three different conditions of controlled atmosphere (CA1, CA2, CA3 and Control) was studied. After 14, 21 and 28 days of cold storage, samples were withdrawn from CA and kept for 4 days in ambient air for ripening. The sensory profile of the peaches and the descriptive terminology were developed by methodology based on the Quantitative Descriptive Analysis (QDA). The panelists consensually defined the sensory descriptors, their respective reference materials and the descriptive evaluation ballot. Fourteen panelists were selected based on their discrimination capacity and reproducibility. Seven descriptors were generated showing similarities and differences between samples. The data were analyzed by ANOVA, Tukey test and Principal Component Analysis (PCA). Results showed significant differences in the sensory profiles of the peaches. The PCA showed that CA2 and CA3 treatments were more characterized by the fresh peach flavor, fresh peach appearance, juiciness and flesh firmness, and were effective in keeping the good quality of the 'Douradão' peaches during the 28 days of cold storage. The Control and CA1 treatments were characterized by the mealiness and were ineffective for quality maintenance of the fruits during cold storage.

Determination of presence and quantification of cadmium, lead and copper in Nile tilapia (Oreochromis niloticus) fillets obtained from three cold storage plants in the state of Parana, Brazil

Pisciculture is an economic activity that is steadily growing in the state of Parana, Brazil, and Nile tilapia (Oreochromis niloticus) is one of the widely cultivated species in this state. Tilapia is not only a very nutritious food, but also an important indicator of environmental contamination. This study aimed to verify contamination by cadmium, copper and lead in tilapia fillets, and to compare the found values to international legislations. Were collected 135 samples of tilapia fillets, between July 2006 and May 2007, in three fish stores located in regions west and north of Paraná State. Samples of tilapia fillet were analyzed in relation to the presence of cadmiun, lead and copper, using atomic absorption spectrophotometry. Lead has not been detected in the analyses. Cadmium has been detected in three samples, on concentrations of 0.012 µg.g-1, 0.011 µg.g-1 and 0.014 µg.g-1. Copper has been detected in all fillets, and the average concentration of each cold storage plant was of 0.122 µg.g-1, 0.106 µg.g-1 and 0.153 µg.g-1. The concentrations found in this study are within the limits allowed by both the European and the Australian legislations.

Intelligence acquisition methods in cyber domain : examining the circumstantial applicability of cyber intelligence acquisition methods using a hierarchical model

Phenomena in cyber domain, especially threats to security and privacy, have proven an increasingly heated topic addressed by different writers and scholars at an increasing pace – both nationally and internationally. However little public research has been done on the subject of cyber intelligence. The main research question of the thesis was: To what extent is the applicability of cyber intelligence acquisition methods circumstantial? The study was conducted in sequential a manner, starting with defining the concept of intelligence in cyber domain and identifying its key attributes, followed by identifying the range of intelligence methods in cyber domain, criteria influencing their applicability, and types of operatives utilizing cyber intelligence. The methods and criteria were refined into a hierarchical model. The existing conceptions of cyber intelligence were mapped through an extensive literature study on a wide variety of sources. The established understanding was further developed through 15 semi-structured interviews with experts of different backgrounds, whose wide range of points of view proved to substantially enhance the perspective on the subject. Four of the interviewed experts participated in a relatively extensive survey based on the constructed hierarchical model on cyber intelligence that was formulated in to an AHP hierarchy and executed in the Expert Choice Comparion online application. It was concluded that Intelligence in cyber domain is an endorsing, cross-cutting intelligence discipline that adds value to all aspects of conventional intelligence and furthermore that it bears a substantial amount of characteristic traits – both advantageous and disadvantageous – and furthermore that the applicability of cyber intelligence methods is partly circumstantially limited.

Determination of some quality properties of marinated sea bream (Sparus Aurata L., 1758) during cold storage

In this study, it was aimed to determine the effect of sea bream (Sparus aurata) marinated some quality properties during cold storage. The fillets of fish were immersed into brine including 3.5% acetic acid 11% salt in the ratio of 1: 1.5 (fish: marinate brine) for marination process. After the process of ripening, samples were grouped into two and packed in plastic containers; one being plain (in sunflower oil) and the other being sauced (sauced prepared with sunflower oil). During storage, sensory, crude protein, lipid, dry matter and crude ash, TBA, TVB-N, TMA-N and peroxide analyses were done periodically. According to results of 200 days of storage, TVB-N values of sea bream marinates packaged as plain and sauced were 15.86/14.89 mg/100g, TBA 7.06/7.99 mg MA/kg, TMA-N 2.97/3.12, mg/100g, the value of peroxide was 7.23/7.45 meq/kg respectively. According to chemical and sensory analyses results obtained in the study; it was concluded that sea bream marinates packaged as plain and sauced can be stored in +4 °C for 200 days.

Microbiological and sensory evaluation of Jambu (Acmella oleracea L.) dried by cold air circulation

Abstract The aim of this study was to evaluate the microbiological and sensory quality of the Jambu (Acmella oleracea L.) in natura and dried by cold air, and the determination of its drying curve. The microbiological analysis were performed to Salmonella spp, the coagulase-positive Staphylococcus, and coliforms in the both Jambu samples, at 45 °C. Tacacá, the typical food dish of Pará state, Brazil, has showed good consumer global acceptance in the sensory evaluation of Jambu in natura (score of 8.00 ± 1.46) and dried (score of 8.67 ± 0.66). Both samples, Jambu in natura and dried by cold air, were by the current legislation regarding the microbiological aspects, this is the absence of Salmonella spp, coagulase-positive Staphylococcus <1×101 CFU/g, and coliforms <3 MPN/g, at 45 °C. Thus, considering sensory and health aspects, the commercialization of dried Jambu becomes viable, facilitating its transportation and handling, as well as for reducing its vegetable mass.