Comparative analysis of methods for assessing the safety of Dunaliella salina Teodoresco and Chlorococcum dissectum Korshikov (Chlorophyta) microalgae cultures after exposure to stress factors

The possibility of using various methods for determining the viability of cultures of microalgae Dunaliella salina and Chlorococcum dissectum before and after freezing-warming was investigated and analyzed. It has been established that the selection of an effective method should be carried out individually for each culture. For an integral assessment of the proliferative and metabolic activity of cells of both species of the studied microalgae, Alamar Blue-test and the ability to grow on liquid nutrient media can be used. The use of the Koch plate method, MTT-test and TTC staining is possible only for the microalga C. dissectum. Vital staining with trypan blue was found to be incorrect.

The Mycorrizal Status in Vineyards Affected by Esca

In this work we analyzed the relationship among native arbuscular mycorrhizal fungi (AMF) and vine roots affected by esca, a serious grapevine trunk disease. The AMF symbiosis was analyzed on the roots of neighboring plants (symptomatic and asymptomatic to esca) in 14 sites of three vineyards in Marche region (central–eastern Italy). The AMF colonization intensity, identified by non-vital staining, showed higher value in all esca symptomatic plants (ranging from 24.6% to 61.3%) than neighboring asymptomatic plants (from 17.4% to 57.6%). The same trend of Glomeromycota phylum abundance was detected by analyzing fungal operational taxonomic units (OTUs) linked to the AMF community, obtained by amplicon high throughput analysis of ITS 1 region. Overall, the highest amount of OTUs was detected on roots from symptomatic plants (0.42%), compared to asymptomatic roots (0.29%). Specific primer pairs for native Rhizophagus irregularis and Funneliformis mosseae AMF species, were designed in 28S rRNA and large subunit (LSU) ribosomal RNA, respectively, and droplet digital PCR protocol for absolute quantification was set up. A higher number of DNA copies of both fungal species were detected more frequently in symptomatic than asymptomatic vines. Our study suggests a relationship between esca and native AMF in grapevine. These results underline the importance of native rhizosphere microbial communities for a better knowledge of grapevine esca disease.

Morphology of the Spleen in Oreochromis niloticus: Splenic Subregions and the Blood-Spleen Barrier

The spleen is a separate organ of the teleost, playing an essential role in immune reactions. The morphology of the spleen is different from the fish species. Little knowledge about the spleen structure and the blood splenic barrier (BSB) in Nile tilapia has been reported. To address this issue, we studied the histology of the spleen and the BSB in healthy Nile tilapia. The morphology of the spleen was observed, then H&E staining, modified Jame’s staining, and ultrastructural techniques were performed to portion the spleen into three subregions and analyze the location of components and fibers. Thereafter, vital staining of Nile tilapia with Trypan blue was conducted to elucidate the composition and function of BSB. Histologically, the spleen could be divided into three subregions (inner, middle, and outer). The venules, clumps of lymphocytes, and vessels were separately characterized features of the outer, middle, and inner layers. Post injection, Trypan blue was intercepted in the endotheliocytes of ellipsoids in the middle layer (i.p.) or was deposited to the reticular fibers surrounding the ellipsoids (i.v.). Additionally, the amount of Trypan blue was shown to be positively correlated to that of the Acid phosphatase expressed. In conclusion, the spleen could be portioned into three subregions, and the BSB lay in the middle layer, composed of the cuboidal-shaped endotheliocytes and the surrounding reticular fibers of the ellipsoid capillaries. The present study enriched the research of immune tissues and system in tilapia and provided reference for the study of spleen in other fish species.

LASIK and Dry Eye Disease: A Case Report

Background: Post- Laser- assisted in situ keratomileusis (LASIK) dry eye is one of the most common postoperative dry eyes after ophthalmic surgeries. Clinically, patients have positive vital staining of the ocular surface, decreased tear breakup times, reduced corneal sensitivity, and decreased visual acuity. In majority of patients, symptoms last about one month, but some patients can experience these symptoms for more than a year. Case presentation: A 29-year-old female with Dry Eye Disease (DED) secondary to isotretinoin use wanted to undergo LASIK for refractive improvement. Ophthalmologists were thorough in informing the patient of the significant risk of post-operative dry eyes, especially in patients with pre-operative DED. A cautious approach was taken by the ophthalmologist, and the patient’s opinion and needs considered. Post-operatively, the patient continues to have DED but reports it is not worse than pre-LASIK and continues to be complaint with dry eye disease treatment. Conclusions: In patients with existing DED, clinicians are vigilant in progressing with LASIK, due to post-operative dry eye symptoms. Careful patient selection and preoperative ocular surface management is recommended for such patient groups. This case reports highlights the importance of clinicians allowing patients to come to an informed decision regarding their care. This patient-centered approach is a model of care that respects patients’ experiences, values, needs and preferences, whilst improving clinical outcomes, better use of resources and increased satisfaction with care.

LASIK and Dry Eye Disease: A Case Report

Background: Post- Laser- assisted in situ keratomileusis (LASIK) dry eye is one of the most common postoperative dry eyes after ophthalmic surgeries. Clinically, patients have positive vital staining of the ocular surface, decreased tear breakup times, reduced corneal sensitivity, and decreased visual acuity. In majority of patients, symptoms last about one month, but some patients can experience these symptoms for more than a year. Case presentation: A 29-year-old female with Dry Eye Disease (DED) secondary to isotretinoin use wanted to undergo LASIK for refractive improvement. Ophthalmologists were thorough in informing the patient of the significant risk of post-operative dry eyes, especially in patients with pre-operative DED. A cautious approach was taken by the ophthalmologist, and the patient’s opinion and needs considered. Post-operatively, the patient continues to have DED but reports it is not worse than pre-LASIK and continues to be complaint with dry eye disease treatment. Conclusions: In patients with existing DED, clinicians are vigilant in progressing with LASIK, due to post-operative dry eye symptoms. Careful patient selection and preoperative ocular surface management is recommended for such patient groups. This case reports highlights the importance of clinicians allowing patients to come to an informed decision regarding their care. This patient-centered approach is a model of care that respects patients’ experiences, values, needs and preferences, whilst improving clinical outcomes, better use of resources and increased satisfaction with care.

Partial Decellularization for Segmental Tracheal Scaffold Tissue Engineering: A Preliminary Study in Rabbits

In this study, we developed a new procedure for the rapid partial decellularization of the harvested trachea. Partial decellularization was performed using a combination of detergent and sonication to completely remove the epithelial layers outside of the cartilage ring. The post-decellularized tracheal segments were assessed with vital staining, which showed that the core cartilage cells remarkably remained intact while the cells outside of the cartilage were no longer viable. The ability of the decellularized tracheal segments to evade immune rejection was evaluated through heterotopic implantation of the segments into the chest muscle of rabbits without any immunosuppressive therapy, which demonstrated no evidence of severe rejection or tissue necrosis under H&E staining, as well as the mechanical stability under stress-pressure testing. Finally, orthotopic transplantation of partially decellularized trachea with no immunosuppression treatment resulted in 2 months of survival in two rabbits and one long-term survival (2 years) in one rabbit. Through evaluations of posttransplantation histology and endoscopy, we confirmed that our partial decellularization method could be a potential method of producing low-immunogenic cartilage scaffolds with viable, functional core cartilage cells that can achieve long-term survival after in vivo transplantation.

Noninvasive Imaging Methods to Improve the Diagnosis of Oral Carcinoma and Its Precursors: State of the Art and Proposal of a Three-Step Diagnostic Process

Oral squamous cell carcinoma (OSCC) is the most prevalent form of cancer of lips and oral cavity, and its diagnostic delay, caused by misdiagnosis at the early stages, is responsible for high mortality ratios. Biopsy and histopathological assessment are the gold standards for OSCC diagnosis, but they are time-consuming, invasive, and do not always enable the patient’s compliance, mainly in cases of follow-up with the need for more biopsies. The use of adjunctive noninvasive imaging techniques improves the diagnostic approach, making it faster and better accepted by patients. The present review aims to focus on the most consolidated diagnostic techniques, such as vital staining and tissue autofluorescence, and to report the potential role of some of the most promising innovative techniques, such as narrow-band imaging, high-frequency ultrasounds, optical coherence tomography, and in vivo confocal microscopy. According to their contribution to OSCC diagnosis, an ideal three-step diagnostic procedure is proposed, to make the diagnostic path faster, better, and more accurate.