Nivin Sharawy

Vasoplegia in septic shock: do we really fight the right enemy?

Vasoplegia is a key factor for the death of patients with septic shock in intensive care unit owing to persistent and irreversible hypotension. Impairment of vascular reactivity has been attributed to a combination of endothelial injury, arginine-vasopressin system dysfunction, release of other vasodilatory inflammatory mediators, and muscle hyperpolarizaton. Nitric oxide induced by a Ca(+2) independent isoform of nitric oxide synthase has been suggested to play an important role in sepsis-induced vasoplegia. However, inhibition of nitric oxide synthase only partially restores the endotoxin-induced vascular hyporeactivity. The aim of this review is to discuss in detail the recent suggested alternative mechanisms of vasoplegia and to briefly outline the current therapeutic strategies and the novel therapeutic options based on those mechanisms.

Estradiol receptors agonists induced effects in rat intestinal microcirculation during sepsis.

The steroid hormone estradiol is suggested to play a protective role in intestinal injury during systemic inflammation (sepsis). Our aim was to determine the effects of specific estradiol receptor (ER-α and ER-ß) agonists on the intestinal microcirculation during experimental sepsis. Male and sham ovariectomized female rats were subjected to sham colon ascendens stent peritonitis (CASP), and they were compared to male and ovariectomized female rats underwent CASP and either estradiol receptor α (ER-α) agonist propyl pyrazole triol (PPT), estradiol receptor ß (ER-ß) agonist diarylpropiolnitrile (DPN), or vehicle treatment. Intravital microscopy was performed, which is sufficiently sensitive to measure changes in the functional capillary density (FCD) as well as the major steps in leukocyte recruitment (rolling and adhesion). The leukocyte extravasations were also quantified by using histological paraffin sections of formalin fixed intestine. We found that either DPN (ER-β) or PPT (ER-α) significantly reduced (P<0.05) sepsis-induced leukocyte-endothelial interaction (rolling, adherent leukocytes and neutrophil extravasations) and improved the intestinal muscular FCD. [PPT: Female; Leukocyte rolling (n/min): V(3) 3.7±0.7 vs 0.8±0.2, Leukocyte adhesion(n/mm(2)): V(3) 131.3±22.6 vs 57.2±13.5, Neutrophil extravasations (n/10000 μm(2)): 3.1±0.7 vs 6 ±1. Male; Leukocyte adhesion (n/mm(2)): V(1) 154.8±19.2 vs 81.3±11.2, V(3) 115.5±23.1 vs 37.8±12]. [DPN: Female; neutrophil extravasations (n/10000 μm(2)) 3.8±0.6 vs 6 ±1. Male; Leukocyte adhesion (n/mm(2)) V(1) 154.8±19.2 vs 70±10.5, V(3) 115.5±23.1 vs 52.8±9.6].Those results suggest that the observed effects of estradiol receptors on different phases of leukocytes recruitment with the improvement of the functional capillary density could partially explain the previous demonstrated salutary effects of estradiol on the intestinal microcirculation during sepsis. The observed activity of this class of compounds could open up a new avenue of research into the potential treatment of sepsis.

Evaluation of the effects of gender and estradiol treatment on the intestinal microcirculation during experimental sepsis

Intestinal barrier dysfunction plays an important role in sepsis. Females may tolerate sepsis better than males, which could be due to the relative resistance of the female intestine to gut injury and inflammation when subjected to sepsis. In this study the intestinal microcirculation was investigated in 50 female and 40 male rats divided in to 9 groups of 10 animals. Male and female rats were subjected to sham CASP (colon ascendens stent peritonitis). We induced experimental sepsis (CASP) in another two groups of male and female rats. The role of the estradiol treatment was evaluated both in male and ovariectomized female rats. Female rats were subjected to sham ovariectomy 3 weeks before sham CASP. Male and ovariectomized female rats were treated with estradiol (10mg/kg estradiol in rizinus oil immediately and 12h following CASP). Another two groups of male and ovariectomized female rats received placebo oil treatment. To evaluate the effects of gender and estradiol treatment on the microvascular perfusion during sepsis, intravital microscopy was performed twenty-four hours after sham CASP or CASP surgery, which permits the in vivo determination of leukocyte-endothelial cell interaction (rolling leukocytes and adherent leukocytes) and the measurement of functional capillary density (FCD), which served as the measure of quality of microvascular perfusion. We found that there was gender difference mainly in the leukocyte endothelial interaction rather than the functional capillary density (FCD), in which male showed significant increases (P<0.05) both in the leukocyte adhesion and rolling leukocytes in submucosal venules (V1 and V3) in comparison to female rats. (Leukocyte adhesion: V1 107.1 ± 49.2n/mm(2); V3 112.3 ± 68.1n/mm(2)) (Rolling leukocytes:V1 16.2 ± 10.3n/min; V3 8.4 ± 8.2n/min). In addition estradiol replacement in ovariectomized female and male rats induced significant decreases (P<0.05) in the leukocyte adhesion and rolling (V1 and V3) with a significant increase in the muscular FCD in comparison to the corresponding placebo treated groups. (Leukocyte adhesion: V1 60 ± 31 n/mm(2); V3 78.11 ± 37.6n/mm(2)) (Rolling leukocytes: V1 13.4 ± 8.9 n/min; V3 5.8 ± 7.4n/min) (Longitudinal muscular FCD (cm/cm(2)): in ovariectomized female rats 107.1 ± 12.2; in male rats 106.2 ± 15.3) (Circular muscular FCD (cm/cm(2)): in ovariectomized female rats: 84.8 ± 14.2; in male rats: 86.1 ± 15.3). We conclude that gender difference in the leukocyte endothelial interaction could explain the resistance of female intestine to injury, and that estradiol treatment could improve the intestinal microcirculation through its effects on the leukocyte endothelial interaction and the FCD both in male and ovariectomized female rats.

New directions for sepsis and septic shock research

BACKGROUND Septic shock is a frequent complication in intensive care unit that can result in multiple organ failure and death. In addition, recent data suggested that severe sepsis and septic shock represent an economic burden. Therefore, septic shock is an important public health problem. METHOD In this review, we will focus on the recent evidences concerning the stages of septic shock, the complex macrocirculation and microcirculation relationship, and the importance of those evidences for future resuscitation goals and therapeutic strategies during late septic shock. RESULT Recently, two stages of septic shock are suggested. In early stage, hypovolemia is the main contributing factor. During this stage, macrocirculatory and microcirculatory changes run parallel, and fluid resuscitation seems to be effective in restoring the hemodynamic parameters. Late stage of septic shock is characterized by complex microcirculation and macrocirculation relationship. CONCLUSIONS Although early goal-directed therapy is a stepwise approach in the treatment of septic shock, tissue perfusion remains an important factor that contributes to septic shock outcome. Because appropriate monitoring of tissue perfusion is a matter of debt, the ideal therapeutic strategy remains a controversial issue that needs further investigations.

Comparative Neuroprotective Effects of Dexamethasone and Minocycline during Hepatic Encephalopathy.

Objective. Encephalopathy and brain edema are serious complications of acute liver injury and may lead to rapid death of patients. The present study was designed to investigate the role of the inflammatory mediators and oxidative stress in the cytotoxic brain oedema and the neuroprotective effects of both minocycline and dexamethasone. Methods. 48 male albino rats were divided into 4 groups: control group, acute liver injury (ALI) group, minocycline pretreated ALI group, and dexamethasone pretreated ALI group. 24 hours after acute liver injury serum ammonia, liver enzymes, brain levels of heme oxygenase-1 gene, iNOS gene expression, nitrite/nitrate, and cytokines were measured. In addition, the grades of encephalopathy and brain water content were assessed. Results. ALI was associated with significant increases in all measured inflammatory mediators, oxidative stress, iNOS gene expression, and nitrite/nitrate. Both minocycline and dexamethasone significantly modulated the inflammatory changes and the oxidative/nitrosative stress associated with ALI. However, only minocycline but not dexamethasone significantly reduced the cytotoxic brain oedema. Conclusion. Both minocycline and dexamethasone could modulate inflammatory and oxidative changes observed in brain after ALI and could be novel preventative therapy for hepatic encephalopathy episodes.

Clinical relevance of early sublingual microcirculation monitoring in septic shock patients

INTRODUCTION Although microcirculation dysfunction plays unique role in septic shock, translation of microcirculation to clinical practices is limited by current semi-quantities analysis and unclear clinical relevance of microcirculation monitoring. Our aim was to critically evaluate the characteristic nature and relevant clinical important of microcirculation. EVIDENCE ACQUISITION Pubmed (2000 to August 2015) were searched to identify observation, case-control, intervention and randomized clinical studies evaluating the relationship between microcirculation alterations and mortality, morbidity and drug responses. The STROBE and CONSORT Statement for assessment of the quality of included studies. EVIDENCE SYNTHESIS We examined results from 17 observations, 4 randomized controlled trials and one case report published studies. This data set comprised of 637 patients. Early septic shock is associated with hypoperfusion and heterogeneous microcirculation that is associated with hyperlactemia and metabolic acidosis. The evidence on clinical relevance of microcirculation is less striking, mainly due to the limited number of studies and problems related to the methodological protocol of the studies and currently semi-quantitative analysis technique. In particular the baseline and time course of microcirculation alteration appears to be controversial. CONCLUSION There is lack of evidences of clinical importance of early microcirculation monitoring and mechanism of microcirculation dysfunction in septic shock patients. This could be due to the methodological protocol of the studies and currently semi-quantitative analysis technique.

Evaluation of the effects of Eserine and JWH-133 on brain dysfunction associated with experimental endotoxemia

UNLABELLED Sepsis is associated with neuronal damage and cognitive impairment, with the participation of pro-inflammatory cytokines and oxidative-nitrous stress. It is known that activated microglia plays a vital role in neuro-inflammation and neuro-degeneration. Thus, the objective of this study was to evaluate therapeutic roles of two microglia regulating agents, JWH-133 and Eserine, on the neuroinflammatory associated brain dysfunctions. To achieve our aim, we used control rats or submitted rats to lipopolysaccharide (LPS) challenge. 30 min after LPS challenge, the animals received either saline, Eserine, JWH-133 or Eserine+JWH-133. After 24h, animals were submitted to the habituation to T maze, Rotarod and activity cage tests. The rats were killed after and were evaluated for central and peripheral inflammatory and oxidative parameters. We observed that the use of Eserine, JWH-133 or Eserine + JWH-133 reverted the increases in the inflammatory markers [interleukin 6 (IL6), vascular cell adhesion molecule 1(VCAM-1) and Eselectin] and oxidative-nitrous stress MDM, and that the anti-inflammatory, antioxidant properties of both JWH-133 and Eserine successfully improve the LPS induced brain dysfunction. CONCLUSIONS The results observed in this study reinforce the role of microglia activation regulating agents, in particular, JWH-133 and Eserine, in the brain dysfunction associated with endotoxemia.

Preliminary clinical evaluation of automated analysis of the sublingual microcirculation in the assessment of patients with septic shock: Comparison of automated versus semi-automated software

INTRODUCTION The outcome of patients in septic shock has been shown to be related to changes within the microcirculation. Modern imaging technologies are available to generate high resolution video recordings of the microcirculation in humans. However, evaluation of the microcirculation is not yet implemented in the routine clinical monitoring of critically ill patients. This is mainly due to large amount of time and user interaction required by the current video analysis software. The aim of this study was to validate a newly developed automated method (CCTools®) for microcirculatory analysis of sublingual capillary perfusion in septic patients in comparison to standard semi-automated software (AVA3®). METHODS 204 videos from 47 patients were recorded using incident dark field (IDF) imaging. Total vessel density (TVD), proportion of perfused vessels (PPV), perfused vessel density (PVD), microvascular flow index (MFI) and heterogeneity index (HI) were measured using AVA3® and CCTools®. RESULTS Significant differences between the numeric results obtained by the two different software packages were observed. The values for TVD, PVD and MFI were statistically related though. CONCLUSION The automated software technique successes to show septic shock induced microcirculation alterations in near real time. However, we found wide degrees of agreement between AVA3® and CCTools® values due to several technical factors that should be considered in the future studies.

Microcirculation research in community hospitals – challenges and chances

Community hospitals provide ideal conditions for large clinical studies because of the high volume of unselected patients admitted every year. With regard to microcirculatory studies, there are still some feasibility problems which are not solved yet. First of all, the lack of reliable automated software to analyze microcirculatory images represents the most important issue. Secondly, hardware aspects still need improvements regarding portability and miniaturization. Finally, to conduct studies of the microcirculation in a community hospital is also always a funding issue. The cost of the measurement device is hereby only one factor. Main cost factor is the personnel.

Tetrahydrobiopterin improves microcirculation in experimental sepsis

BACKGROUND Tetrahydrobiopterin (BH4), an endogenous nucleic acid derivative, acts as an important cofactor for several enzymes found within the vascular endothelium, which is deranged in sepsis. OBJECTIVE We hypothesized that BH4 would improve capillary density and decrease inflammation within the intestinal microcirculation of septic rats. METHODS We conducted a randomized, controlled trial using two previously validated models of sepsis in rats: 1) A fecal peritonitis model using a stent perforating the ascending colon, and 2) An endotoxemia model using lipopolysaccharide (LPS) toxin from E. coli. Experimental groups receiving BH4 (60 mg/kg) were compared to otherwise healthy controls and to untreated groups with sepsis-like physiology. RESULTS BH4 decreased leukocyte-endothelial adhesion by 55% and 58% (P < 0.05) in the peritonitis model and endotoxemia models, respectively. In the endotoxemia model but not the peritonitis model, BH4 improved functional capillary density in capillary beds within the intestine (141.3 vs. 106.7 mm/cm2, p < 0.05). Macrohemodynamic parameters were no different between placebo treatment and BH4-treated groups. CONCLUSIONS This study demonstrates that BH4 improves capillary density and inflammation in two separate models of sepsis. BH4 may represent a novel adjunct in the treatment of sepsis and septic shock in clinical practice. Further dose-finding studies and clinical trials are warranted.