Cannabidiol nanoemulsion for eye treatment - Anti-inflammatory, wound healing activity and its bioavailability using in vitro human corneal substitute.

Cannabidiol (CBD) is the non-psychoactive component of the plant Cannabis sativa (L.) that has great anti-inflammatory benefits and wound healing effects. However, its high lipophilicity, chemical instability, and extensive metabolism impair its bioavailability and clinical use. Here, we report on the preparation of a human cornea substitute in vitro and validate this substitute for the evaluation of drug penetration. CBD nanoemulsion was developed and evaluated for stability and biological activity. The physicochemical properties of CBD nanoemulsion were maintained during storage for 90 days under room conditions. In the scratch assay, nanoformulation showed significantly ameliorated wound closure rates compared to the control and pure CBD. Due to the lower cytotoxicity of nanoformulated CBD, a higher anti-inflammatory activity was demonstrated. Neither nanoemulsion nor pure CBD can penetrate the cornea after the four-hour apical treatment. For nanoemulsion, 94 % of the initial amount of CBD remained in the apical compartment while only 54 % of the original amount of pure CBD was detected in the apical medium, and 7 % in the cornea, the rest was most likely metabolized. In summary, the nanoemulsion developed in this study enhanced the stability and biological activity of CBD.

FORMS OF OCULAR LARVAL TOXOCARIASIS IN CHILDHOOD. A REVIEW.

The aim of this paper is to present an outline of forms of ocular manifestations of visceral larva migrans in children, as illustrated by the extensive photographic documentation. Ocular larval toxocariasis (OLT) has various clinical manifestations even in childhood age, in which age representation has an influence. The most common is presence of peripheral granuloma of the eye, frequently with a tractional vitreal streak leading from the retinal periphery to the optic nerve papilla. This is followed by granuloma of the posterior pole of the eye, usually reaching from the macular landscape to the central retinal periphery, always with vitritis. In children OLT may be manifested also in affliction of the optic nerve (cystic granuloma of the head of the optic nerve or neuropathy with vitreal reaction), fulminant endophthalmitis and in rare cases also diffuse chorioretinitis. The diagnosis rests upon a clinical ophthalmological finding, as well as laboratory examination of the levels of antibodies with potential eosinophilia. Histological examination may demonstrate spherical polypoid ossification in the choroid at the posterior pole of the eye as a consequence of fibrotisation and calcification, proceeding from the surrounding area of the absorbed larva. General combined treatment with antihelminthics and corticosteroids is arduous and does not always produce the desired effect in the sense of a satisfactory improvement of visual acuity. In differential diagnostics, manifestations of OLT in small children are still associated with retinoblastoma and a clinical picture of other intraocular diseases.

Vasa nervorum of epicardial nerves and valves of small veins investigated in porcine heart by two types of vascular injections.

The morphology of the vascular supply of peripheral branches of cardiac nerves has not been systematically described until now. The aim of this study was to describe the architectonics of the vasa nervorum of epicardial nerves in porcine hearts by using two injection techniques. Twenty-three hearts from young healthy pigs were used. In 10 hearts India ink solution was injected into the origin of the anterior interventricular branch. In another 10 hearts India ink solution was injected retrogradely through the coronary sinus. The hearts were then analyzed using a magnifying glass and light microscopy. The arterial injection showed the entirety of the rich venous components of the vasa nervorum, which often consisted of paired veins accompanying the epicardial nerves. The thickness of the nerves ranged from 50 to 815 µm. The vasa nervorum drained into larger subepicardial veins. In seven of the hearts prepared with venous injections the vasa nervorum of epicardial nerves were visualized in the same detail as in the arterial preparations and India ink solution filled the right ventricle via the smallest cardiac veins. The histological analysis of these seven hearts showed complete dehiscence and functional insufficiency of small and larger veins valves. In the other three hearts prepared with venous injections the valves were competent, which prevented retrograde filling of larger and smaller veins. The results obtained expand the current knowledge on epicardial nerves vasa nervorum and provide anatomical evidence behind the mechanism of retrograde application of cardioplegic solutions in cardiac surgery.

Vascular supply of the anterior interventricular epicardial nerves and ventricular Purkinje fibers in the porcine hearts.

The aim of this study was to perform a pilot histological and quantitative analysis of the blood vessels accompanying the epicardial nerves (vasa nervorum) in the porcine hearts. Twenty healthy porcine hearts were used in this study. The blood vessels were analyzed by light microscopy using four different staining techniques in transverse sections taken from the upper, middle, and lower segments of the anterior part of the interventricular region and the adjacent parts of the right and left ventricles containing epicardial nerves and the endocardial peripheral parts of the Purkinje fibers. In total, 317 epicardial nerves were detected. The vasa nervorum were present in 75.7% of these nerves. The vasa nervorum resembled arterioles and postcapillary and collecting venules. One hundred and forty nine epicardial nerves were perivascular, located in the adventitia of the anterior interventricular artery and vein. The remaining 168 nerves ran freely through the epicardial interstitium. The presence of the vasa nervorum was not related to topographical location or nerve diameter. Additionally, from a total of 33 analyzed ventricular complexes of Purkinje fibers small blood vessels located in their proximity were identified in only two cases. It can be concluded that the majority of the anterior epicardial nerves of porcine heart possess well-developed vasa nervorum. In contrast, similar blood vessels are rarely present in the vicinity of the Purkinje fibers. The data obtained contribute to a better understanding of the nutrition of the cardiac nerves.

Valves of the small coronary veins in porcine hearts.

The aim of the study was to investigate the existence of valves in small peripheral coronary veins of porcine hearts. The study was performed on 20 porcine hearts using standard histological methods. The veins in the subepicardial and intramyocardial regions of the anterior and posterior parts of the interventricular septum and in the wall of the right atrium were studied. Valves were present in intramyocardial veins (diameter of 75-180 µm), in the veins located just beneath the external surface of the myocardium (diameter 120-170 µm) and in the terminal segments of the ventricular veins (diameter 250 µm) opening into the stems of the anterior interventricular vein and middle cardiac vein. Valves were also recorded in most veins of the subepicardial space. The described rich presence of valves in the small coronary veins may contribute to a better comprehension of their hemodynamic properties. These findings may also help to improve the understanding of the efficacy of retrograde application of medications, a novel technique in cardiology and cardiac surgery.

Morphology of the vasa vasorum in coronary arteries of the porcine heart: A new insight.

INTRODUCTION: The vasa vasorum interna were described during the last decade as a special kind of vessels originating directly from the lumen of the paternal artery and participating in the nourishment of its wall, especially of the aorta and coronary arteries. At the same time, their existence was repeatedly denied/negated by many other authors. AIM: The purpose of the actual study was the anatomical verification of the existence of the vasa vasorum interna in porcine coronary arteries. MATERIALS AND METHODS: The vascular supply was studied on the wall of the anterior interventricular branch of the left coronary artery on 36 hearts taken from healthy pigs. Light microscopy, vascular injections and scanning electron microscopy were used for the analysis of 141 samples. RESULTS: In only two cases small arteries resembling vasa vasorum interna and originating directly from the lumen of the coronary artery were found. But, in both cases these vessels ran without branching, passed over the whole thickness of adventitia and branched in the wider periarterial space. In contrast to this all feeding arteries of the vasa vasorum arose from the larger branches of the paternal artery, branched entirely in its adventitia and did not enter the media. CONCLUSION: Due to the very low incidence of these small arteries originating from the lumen of the paternal artery and the absence of their participation on the nourishment of the arterial wall we came to the conclusion that it is not suitable to use the term "vasa vasorum interna" for their designation.

Vasa vasorum: an old term with new problems.

PURPOSE: The aim of the study was to describe the origin of the Latin anatomical term vasa vasorum and its role in current medical research and to present examples of grammatical errors in its use. METHODS: Literary searches oriented on the term vasa vasorum were used to identify publications using it in the medical literature from the seventeenth century up to the present. RESULTS: The Latin term vasa vasorum was introduced by Ludwig in 1739. The vasa vasorum became an important topic in clinical research around the middle of the twentieth century, with implications in angiology, cardiology and cardiosurgery. We report 18 grammatical errors concerning the use of the term vasa vasorum, starting from the year 1959. A similar decline in the correct use of Latin terminology is also evident in other medical research disciplines. CONCLUSIONS: The numerous errors found in the use of Latin terminology in recent medical literature have occurred as a consequence of decreased use of Latin in the medical community. The only way to improve this situation is by improving awareness of international standard anatomical terminology, which is available worldwide in both Latin and English.

Vasa vasorum of the failed aorto-coronary venous grafts.

PURPOSE: This study was designed to investigate the distribution of vasa vasorum in walls of failed aorto-coronary venous grafts. METHODS: Fifty-one diseased venous grafts harvested from 39 patients underwent qualitative histological evaluation. The morphology of the grade of the pathological changes and the extent of the vascularisation were examined, and related to the length of the interval between the primary surgery and the explantation. The obtained results were placed into five groups, substantially differing one from the other in morphology and vascularisation. RESULTS: The intervals between grafts implantation and explantation ranged from 1 day to 35 years. The onset of arterialization of the graft media was observed on average at 1 month after bypass implantation. During this same time period massive intimal hyperplasia and atherosclerosis occurred. Vasa vasorum proliferation from the adventitia to the outer layers of the media was first apparent between 7 and 24 months after implantation. Proliferation of the vasa vasorum throughout the entire atherosclerotic media and hyperplastic intima continued for a much longer time interval. CONCLUSION: No correlation between neoangiogenesis and age, sex or type of bypassed coronary branch was proven. Regarding the given findings, the authors believe that changes in hemodynamic conditions and endothelial trauma are primarily responsible for the development of graft disease and that vasa vasorum proliferation is only a secondary reaction to the structural changes of the graft wall. To what extent the frequently present pre-existing intimal hyperplasia of venous bypass grafts play in the development of graft disease remains questionable.

Ectopic GRHL2 Expression Due to Non-coding Mutations Promotes Cell State Transition and Causes Posterior Polymorphous Corneal Dystrophy 4.

In a large family of Czech origin, we mapped a locus for an autosomal-dominant corneal endothelial dystrophy, posterior polymorphous corneal dystrophy 4 (PPCD4), to 8q22.3-q24.12. Whole-genome sequencing identified a unique variant (c.20+544G>T) in this locus, within an intronic regulatory region of GRHL2. Targeted sequencing identified the same variant in three additional previously unsolved PPCD-affected families, including a de novo occurrence that suggests this is a recurrent mutation. Two further unique variants were identified in intron 1 of GRHL2 (c.20+257delT and c.20+133delA) in unrelated PPCD-affected families. GRHL2 is a transcription factor that suppresses epithelial-to-mesenchymal transition (EMT) and is a direct transcriptional repressor of ZEB1. ZEB1 mutations leading to haploinsufficiency cause PPCD3. We previously identified promoter mutations in OVOL2, a gene not normally expressed in the corneal endothelium, as the cause of PPCD1. OVOL2 drives mesenchymal-to-epithelial transition (MET) by directly inhibiting EMT-inducing transcription factors, such as ZEB1. Here, we demonstrate that the GRHL2 regulatory variants identified in PPCD4-affected individuals induce increased transcriptional activity in vitro. Furthermore, although GRHL2 is not expressed in corneal endothelial cells in control tissue, we detected GRHL2 in the corneal "endothelium" in PPCD4 tissue. These cells were also positive for epithelial markers E-Cadherin and Cytokeratin 7, indicating they have transitioned to an epithelial-like cell type. We suggest that mutations inducing MET within the corneal endothelium are a convergent pathogenic mechanism leading to dysfunction of the endothelial barrier and disease.

Opportunistic Infections in HIV-Infected Patients Differ Strongly in Frequencies and Spectra between Patients with Low CD4+ Cell Counts Examined Postmortem and Compensated Patients Examined Antemortem Irrespective of the HAART Era.

OBJECTIVE: AIDS-related mortality has changed dramatically with the onset of highly active antiretroviral therapy (HAART), which has even allowed compensated HIV-infected patients to withdraw from secondary therapy directed against opportunistic pathogens. However, in recently autopsied HIV-infected patients, we observed that associations with a broad spectrum of pathogens remain, although detailed analyses are lacking. Therefore, we focused on the possible frequency and spectrum shifts in pathogens associated with autopsied HIV-infected patients. DESIGN: We hypothesized that the pathogens frequency and spectrum changes found in HIV-infected patients examined postmortem did not recapitulate the changes found previously in HIV-infected patients examined antemortem in both the pre- and post-HAART eras. Because this is the first comprehensive study originating from Central and Eastern Europe, we also compared our data with those obtained in the West and Southwest Europe, USA and Latin America. METHODS: We performed autopsies on 124 HIV-infected patients who died from AIDS or other co-morbidities in the Czech Republic between 1985 and 2014. The pathological findings were retrieved from the full postmortem examinations and autopsy records. RESULTS: We collected a total of 502 host-pathogen records covering 82 pathogen species, a spectrum that did not change according to patients' therapy or since the onset of the epidemics, which can probably be explained by the fact that even recently deceased patients were usually decompensated (in 95% of the cases, the last available CD4+ cell count was falling below 200 cells*µl-1) regardless of the treatment they received. The newly identified pathogen taxa in HIV-infected patients included Acinetobacter calcoaceticus, Aerococcus viridans and Escherichia hermannii. We observed a very limited overlap in both the spectra and frequencies of the pathogen species found postmortem in HIV-infected patients in Europe, the USA and Latin America. CONCLUSIONS: The shifts documented previously in compensated HIV-infected patients examined antemortem in the post-HAART era are not recapitulated in mostly decompensated HIV-infected patients examined postmortem.

Temporal arteritis with scalp ulceration and blindness.

Temporal arteritis or giant cell arteritis is a systemic granulomatous vasculitis of medium and large-sized arteries, most frequently involving the temporal artery. It presents with headache, fever, elevated erythrocyte sedimentation rate and anemia. If untreated, complications like ischemic optic neuritis may even lead to blindness. Although very rare, scalp necrosis and ulcerations are the most common dermatologic findings. An 85-year-old woman presented with a two-month history of bilateral scalp ulcerations, followed by permanent loss of vision in the left eye due to the delay in diagnosing temporal arteritis.

Cribriform adenocarcinoma of the base of the tongue and low-grade, polymorphic adenocarcinomas of the salivary glands.

Low-grade, polymorphic adenocarcinomas occur mainly in females and are usually associated with the small salivary glands of the palate. The tumors are malignant, but not aggressive. Regional neck as well as distant metastasis is rare and the mortality rate is low. Cribriform adenocarcinoma of the salivary glands is a rare tumor, currently ranked among low-grade, polymorphic adenocarcinomas of the salivary glands. However, it differs from carcinomas in this group as it metastasizes to the cervical lymph nodes and exhibits frequent primary localization in the small salivary glands at the base of the tongue. Despite the tendency to metastasize, patient prognosis remains favorable. A case of a 72-year-old woman with neck metastases of cribriform adenocarcinoma, of unknown primary origin, is reported. The primary tumor origin was ultimately determined using nuclear magnetic resonance, histological verification was difficult due to the presence of an intact mucosal cover over the tumor. Cribriform adenocarcinoma is known to have a number of characteristics in common with a typical low-grade, salivary gland adenocarcinoma. However, in contrast to low-grade adenocarcinomas, the tumor presented with neck lymph node metastasis.

The correlation between echographic and histopathological findings in uveal melanoma.

PURPOSE: To correlate between the echographic and histopathological findings, and to search for important criteria for the prognosis in vivo using standardised echography. METHODS: The study included 51 consecutive patients treated by primary enucleation for choroidal or ciliary body melanoma and 30 patients who underwent secondary enucleation after failing radiotherapy treatment. All patients were treated during the period of 1998--2008 at the Eye Clinic of the 3rd Faculty of Medicine in Prague, Czech Republic. Standardised echography based on the Ossoinig method was used in this study. Histopathological processing and evaluation was performed by the Laboratory of Ocular Pathology at the same clinic. Professional statistical software (Statistica 7.1 by Statsoft, Inc.) was used for all statistical analysis. RESULTS: Statistically significant results were found in these correlations in the group of primary enucleation: US prominence vs. HP prominence (R = 0.8943, p = 0.0000), US base vs. HP base (R = 0.8060, p = 0.0000), US tumor shape vs. HP tumor shape (R = -0.7576, p = 0.0000), posterior choroidal spike condition vs. scleral invasion (R = -0.5341, p = 0.0005), TNM vs. pTNM (R = 0.7242, p = 0.0000). In the group of secondary enucleation: US prominence vs. HP prominence (R = 0.8132, p = 0.0000), US base vs. HP base (R = 0.7959, p = 0.0000). Other correlations were statistically insignificant. CONCLUSIONS: Tumor dimensions, shape of the tumor and scleral invasion are important predictors that can be in vivo reliably determined using standardised echography.

Uveal melanoma in congenital ocular melanocytosis mimicking malignant transformation of the optic disc melanocytoma (a controlled case).

A case of a 57-year old woman with uveal melanoma of the posterior pole arising in the congenital ocular melanocytosis mimicking clinically malignant transformation of the optic disc melanocytoma is presented.

[Angiolymphoid hyperplasia with eosinophilia with bilateral involvement of the lacrimal glands]. / Angiolymphoide Hyperplasie mit Eosinophilie mit bilateralem Befall der Tränendrüsen.

BACKGROUND: Angiolymphoid hyperplasia with eosinophilia (ALHE) and Kimura's disease are two rarely occurring disorders very similar to each other; however, they are individual nosological entities. For a long time they were considered as a single disease due to the frequency of similar characteristics. The majority of authors have recently considered these diseases as two individual entities with some reciprocal specifications, both clinical as well as morphological. PATIENT: We report on the case of a 76-year old male white European who had suffered for more than six years from subcutaneous tumour formations in different parts of the body. Eyelid oedema on the right side with palpable resistance under the upper temporal edge of the orbit initially occurred five years after the first symptoms of the disease. A similar finding occurred on the left side after eight months. The tumours on both sides were surgically removed and sent for histological analysis. A unilateral recurrence of the finding appeared after one year, followed by surgical intervention and histological examination. RESULTS: Tumour infiltrate in the first two orbita excisions was topically related to lacrimal gland structures; these structures were not found in the third excision. The case was histologically diagnosed as ALHE despite some similar characteristics with Kimura's disease. The diagnosis of ALHE was histologically supported, especially by the absence of lymphatic follicle formations and fibrotisation in the infiltrate, and the identification of the appearance of epitheloid up to histiocytoid of proliferating endothelium, which forms small lumens. The diagnosis of ALHE was additionally supported by the fact that the incidence of Kimura's disease in white Europeans is very rare. CONCLUSIONS: According to our findings, both entities overlap one another, even in some characteristics considered to be distinguishing. The question arises, therefore, whether the strict separation of these nosological entities can indeed be determined with one hundred percent accuracy.