Toll-Like Receptors -2, -3, -4 and -7 Expression Patterns in the Liver of a CLP-Induced Sepsis Mouse Model.

Objective: To investigate the expression of toll-like receptors (TLRs) in the liver of septic mouse model. Materials and methods: For this study seventy-two C57BL/6J mice were utilized. Sepsis was induced by cecal ligation and puncture (CLP) in the mice of the three septic (S) groups (euthanized at 24 hours, 48 hours and 72 hours). Sham (laparotomy)- operated mice constituted the control (C) groups (euthanized at 24, 48 and 72 hours). Blood samples were drawn and liver tissues were extracted and examined histologically. The expression of TLRs 2, 3, 4 and 7 was assessed via immunohistochemistry (IHC) and qrt-PCR (quantitative- Polymerase Chain Reaction). Results: Liver function tests were elevated in all S-groups in contrast to their time-equivalent control groups (S24 versus C24, S48 versus C48 and S72 versus C72) (p < 0.05). Liver histology displayed progressive deterioration in the septic groups. IHC and qrt-PCR both showed an increased expression of all TLRs in the septic mice in comparison to their analogous control ones (p < 0.05). Analysis of livers and intestines of the septic animals proved that all TLRs were significantly expressed in higher levels in the intestinal tissues at 24h and 48h (p < 0.05) except for TLR 3 in S48 (p > 0.05); whereas at 72 hours only TLR 4 levels were significantly elevated in the intestine (p < 0.05). Conclusion: TLRs seem to be expressed in significant levels in the livers of septic rodents, indicating that they have a possible role in the pathophysiology of liver damage in septic conditions.

Increased expression of Toll-like receptors 2, 3, 4 and 7 mRNA in the kidney and intestine of a septic mouse model.

Toll-like receptors (TLRs) are the key regulators of innate and adaptive immunity and are highly expressed during sepsis. Thus, studying the expression of TLRs in an animal septic model might indicate their possible association with acute kidney injury in sepsis. Seventy-two male C57BL/6J mice were used for this study. Randomly, these animals were divided into 6 groups (N = 12/group): 3 control and 3 septic groups depending on the euthanasia time (24 h, 48 h, 72 h). Septic groups underwent cecal ligation and puncture (CLP) to induce peritonitis, while control groups had a sham operation. Hematological tests were performed in serum for immune biomarkers; immunohistochemistry, morphometry and qRT-PCR analysis were used on both kidney and intestine tissues to evaluate the expression of TLR 2, 3, 4 and 7 in a septic process. At the end of each experimental period, we found that TLRs 2, 3, 4 and 7 were expressed in both tissues but there were differences between those at various time points. Also, we found that mRNA levels were significantly higher in qRT-PCR evaluation in septic groups than control groups in both kidney and intestinal tissues (p < 0.05); showing a steady increase in the septic groups as the time to euthanasia was prolonged (p < 0.05). Overall, our study provides a suggestion that TLRs 2, 3, 4 and 7 are highly expressed in the kidneys of septic mice and especially that these TLRs are sensitive and specific markers for sepsis. Finally, our study supports the diagnostic importance of TLRs in AKI and provides an insight on the contribution of septic mice models in the study of multi organ dysfunction syndrome in general.

Cerebral blood flow velocity alterations, under two different carbon dioxide management strategies, during sevoflurane anesthesia in gynecological laparoscopic surgery.

In this study, 33 female patients, scheduled for operative gynecological laparoscopies, were enrolled. Our aim was prospective, randomized comparison of the influence of two different management strategies, regarding end tidal CO2, on cerebral blood flow velocities and on pulsatility index, examined by means of transcranial Doppler ultrasonography, under sevoflurane anesthesia 1.3 MAC: permissive hypercapnia (up to 45 mmHg, Group I, n = 17) versus intervention to ensure mild hypocapnia, (around 33 mmHg, Group II, n = 16). Baseline measurements of investigated parameters were recorded and CO2 insufflation started. In Group I no further adjustment was performed and CO2 partial pressure rose, while in Group II it was kept stable, by ventilatory patterns adjustment. Hemodynamic, acid base balance and cerebrovascular variables were recorded during pneumoperitoneum and in post-desufflation period, at eight checking time points. In Group I cerebral blood flow velocities increased according to CO2 elevation (2.3%-3.9% per mmHg of increase in CO2 partial pressure), whereas in Group II no significant alterations were noticed. Pulsatility index was constant over time without clinical differences between groups. Our study suggests that under sevoflurane anesthesia 1.3 MAC, prophylactic hyperventilation limits the cerebral blood flow velocities enhancing effect of CO2 insufflation, during laparoscopies.