Role of Corneal Epithelial Measurements in Differentiating Eyes with Stable Keratoconus from Eyes that Are Progressing.

Purpose: To evaluate measures of corneal epithelium in eyes that showed documented signs of keratoconus (KC) progression and compare with stable eyes and healthy controls. Also, to determine the correlation of these epithelial parameters with maximum keratometry (K max) and pachymetry. Design: Prospective, observational, comparative study. Participants: One-hundred and fifty eyes from 150 patients. The study included 50 eyes from patients with documented KC progression, 50 eyes with stable KC, and 50 clinically normal eyes to serve as controls. Methods: A spectral-domain (SD)-OCT imaging was obtained in all eyes, and mean values were compared between the groups. The correlation of epithelial parameters with K max and thinnest pachymetry was also investigated. Main Outcome Measures: For the purposes of this study, the epithelial measures maximum, minimum, superior, and inferior values as well as the difference between the minimum and maximum (min-max) and epithelial standard deviation were considered, obtained from SD-OCT and compared between groups. Measurements of the thinnest point and min-max in pachymetry were also recorded. Results: The only epithelial parameter that presented a statistically significant difference between stable and progressive KC was epithelium min-max. Although stable KC presented epithelium min-max mean values of -18.2 ± 6.6, progressive KC eyes presented mean values of -23.4 ± 10.3 (P < 0.0001). Epithelial maximum (P = 0.16), minimum (P = 0.25), superior (P = 0.28), inferior (P = 0.23), and standard deviation (P = 0.25) values were not significantly different between stable and progressive eyes. Difference min-max pachymetry points in stable (-108.3 ± 33.5) and progressive KC (-115.2 ± 56.0) were not significantly different (P = 0.723). There was no significant correlation between epithelium min-max with corneal thinning (P = 0.39) or K max (P = 0.09) regardless of disease progression. Conclusions: Epithelial measures are useful to identify KC eyes that are progressing; the parameters that measure the difference between min-max epithelium points were significantly different between stable and progressive groups, unlike this difference in pachymetry. Finally, this epithelial parameter seems to be independent of corneal thinning and K max. Financial Disclosures: Proprietary or commercial disclosure may be found after the references.

7-Ketocholesterol Promotes Oxiapoptophagy in Bone Marrow Mesenchymal Stem Cell from Patients with Acute Myeloid Leukemia.

7-Ketocholesterol (7-KC) is a cholesterol oxidation product with several biological functions. 7-KC has the capacity to cause cell death depending on the concentration and specific cell type. Mesenchymal stem cells (MSCs) are multipotent cells with the ability to differentiate into various types of cells, such as osteoblasts and adipocytes, among others. MSCs contribute to the development of a suitable niche for hematopoietic stem cells, and are involved in the development of diseases, such as leukemia, to a yet unknown extent. Here, we describe the effect of 7-KC on the death of bone marrow MSCs from patients with acute myeloid leukemia (LMSCs). LMSCs were less susceptible to the death-promoting effect of 7-KC than other cell types. 7-KC exposure triggered the extrinsic pathway of apoptosis with an increase in activated caspase-8 and caspase-3 activity. Mechanisms other than caspase-dependent pathways were involved. 7-KC increased ROS generation by LMSCs, which was related to decreased cell viability. 7-KC also led to disruption of the cytoskeleton of LMSCs, increased the number of cells in S phase, and decreased the number of cells in the G1/S transition. Autophagosome accumulation was also observed. 7-KC downregulated the SHh protein in LMSCs but did not change the expression of SMO. In conclusion, oxiapoptophagy (OXIdative stress + APOPTOsis + autophagy) seems to be activated by 7-KC in LMSCs. More studies are needed to better understand the role of 7-KC in the death of LMSCs and the possible effects on the SHh pathway.

Interplay between early-life malnutrition, epigenetic modulation of the immune function and liver diseases.

Early-life nutrition plays a critical role in fetal growth and development. Food intake absence and excess are the two main types of energy malnutrition that predispose to the appearance of diseases in adulthood, according to the hypothesis of 'developmental origins of health and disease'. Epidemiological data have shown an association between early-life malnutrition and the metabolic syndrome in later life. Evidence has also demonstrated that nutrition during this period of life can affect the development of the immune system through epigenetic mechanisms. Thus, epigenetics has an essential role in the complex interplay between environmental factors and genetics. Altogether, this leads to the inflammatory response that is commonly seen in non-alcoholic fatty liver disease (NAFLD), the hepatic manifestation of the metabolic syndrome. In conjunction, DNA methylation, covalent modification of histones and the expression of non-coding RNA are the epigenetic phenomena that affect inflammatory processes in the context of NAFLD. Here, we highlight current understanding of the mechanisms underlying developmental programming of NAFLD linked to epigenetic modulation of the immune system and environmental factors, such as malnutrition.

The role of keratinocyte-derived chemokine (KC) on hyperalgesia caused by peripheral nerve injury in mice.

Chemokines are associated with both inflammatory and immune responses and play an important role in the pathophysiological process associated with neuropathic pain following peripheral nerve injury. Here, we investigated the involvement of peripheral keratinocyte-derived chemokine (KC) in the pathogenesis of neuropathic pain induced by the partial ligation of the sciatic nerve (PLSN) in mice. PLSN increased KC levels and its mRNA in both the sciatic nerve and spinal cord when compared with sham-operated mice. In addition, PLSN-induced mechanical and thermal hyperalgesia was prevented by systemic (i.v.) treatment with anti-KC antibody either at the time of surgery or on the 4th day after surgery. Also, intrathecal (i.t.) injection of anti-KC antibody prevented mechanical hyperalgesia induced by PLSN when administered at the time of surgery or on the 4th day after surgery. Importantly, the intraneural (i.n.) injection of KC in the mouse sciatic nerve elicited long-lasting mechanical hyperalgesia, which was prevented by the selective CXCR2 antagonist SB225002. The established mechanical hyperalgesia induced by KC was expressively reduced by the treatment with gabapentin, a drug widely used to treat chronic pain in humans. Intraneural KC injection also caused neutrophil migration into the mouse sciatic nerve and the depletion of neutrophils, by pre-treating animals with vinblastine, significantly reduced KC-induced mechanical hyperalgesia. Similar results were obtained for the pre-treatment with indomethacin, a non-selective COX inhibitor. We also demonstrated an increased level of cytokines (IL-1ß, IL-6, and MCP-1, but not TNF-α) after i.n. injection of KC in the mouse sciatic nerve. Together, these findings suggest a role for KC in the development of neuropathic pain in mice by attracting neutrophils to the injured site and increasing the production of proinflammatory mediators. Therefore, strategies to inhibit the action or the release of this chemokine could constitute a therapeutic tool for the management of neuropathic pain in humans.

A method for retrieving endodontic or atypical nonendodontic separated instruments from the root canal: a report of two cases.

AIM: This clinical report presents a new method for retrieving separated instruments from the root canal with minimally invasive procedures. BACKGROUND: The presence of separated instrument in root canal may interfere in the endodontic treatment prognosis. There are several recommended methods to retrieve separated instruments, but some are difficult in clinically practice. CASE REPORT: This study describes two cases of separated instrument removal from the root canal using a stainless-steel prepared needle associated with a K-file. Case 1 presented a fractured gutta-percha condenser within the mandibular second premolar, it was separated during incorrect intracanal medication calcium hydroxide placement. Case 2 had a fractured sewing needle within the upper central incisor that the patient used to remove food debris from the root canal. After cervical preparation, the fractured instruments were fitted inside a prepared needle and then an endodontic instrument (#25 K-file) was adapted with clockwise turning motion between the needle inner wall and the fragment. CONCLUSION: The endodontic or atypical nonendodontic separated instrument may be easily pull on of the root canal using a single and low cost device. CLINICAL SIGNIFICANCE: The methods for retrieving separated instruments from root canal are difficult and destructive procedures. The present case describes a simple method to solve this problem.

Cholesterol oxidation and astaxanthin degradation in shrimp during sun drying and storage.

Dried salted shrimps are made from raw shrimps, which are cooked and dried under direct sunlight. The preparation and storage include treatments and conditions that can promote oxidative changes in different components. The aim of this study was to monitor the formation of major cholesterol oxidation products and the changes in the astaxanthin content and fatty acid profile in dried salted shrimp during cooking, sun drying and storage. During sun drying, most of the astaxanthin (75%) was degraded in cooked shrimp, while cholesterol oxidation products (COPs) showed a dramatic increase (8.6-fold), reaching a total concentration of 372.9 ± 16.3 µg/g of lipids. Further storage favoured both astaxanthin degradation (83%) and COPs formation (886.6 ± 97.9 µg/g of lipids after 90 days of storage). The high degradation of astaxanthin and the elevated formation of COPs during sun drying and storage indicate the necessity to re-evaluate the processing and storage conditions of salted dried shrimp.

Local inflammatory reaction induced by Scolopendra viridicornis centipede venom in mice.

Centipede envenomation is generally mild, and human victims usually manifest burning pain, erythema and edema. Despite the abundance and ubiquity of these animals, centipede venom has been poorly characterized in literature. For this reason, the aim of this work was to investigate local inflammatory features induced by Scolopendra viridicornis centipede envenomation in mice, evaluating edema formation, leukocyte infiltration, production of inflammatory mediators, and also performing histological analysis. The highest edematogenic activity induced by the venom, determined by plethysmometry, was noticed 0.5 h after injection in mice footpad. At 24 h, edema was still detected in animals that received 15 and 60 µg of venom, and at 48 h, only in animals injected with 60 µg of venom. In relation to leukocyte count, S. viridicornis venom induced cell recruitment, mainly neutrophils and monocytes/macrophages, in all doses and time periods analyzed in comparison with PBS-injected mice. An increase in lymphocytes was detected especially between 1 and 24 h at 60 µg dose. Besides, eosinophil recruitment was observed mainly for 15 and 60 µg doses in early time periods. Edema formation and cell recruitment were also confirmed by histological analysis. Moreover, S. viridicornis venom stimulated the release of IL-6, MCP-1, KC, and IL-1ß. Conversely, S. viridicornis venom did not induce the release of detectable levels of TNF-α. We demonstrated that the edematogenic activity induced by S. viridicornis venom was of rapid onset, and the venom stimulated secretion of pro-inflammatory mediators which contribute to the inflammatory reaction induced by S. viridicornis venom in an experimental model.