The human milk oligosaccharide 3′sialyllactose reduces low-grade inflammation and atherosclerosis development in mice

Macrophages contribute to the induction and resolution of inflammation and play a central role in chronic low-grade inflammation in cardiovascular diseases caused by atherosclerosis. Human milk oligosaccharides (HMOs) are complex unconjugated glycans unique to human milk that benefit infant health and act as innate immune modulators. Here, we identify the HMO 3'sialyllactose (3'SL) as a natural inhibitor of TLR4-induced low-grade inflammation in macrophages and endothelium. Transcriptome analysis in macrophages revealed that 3'SL attenuates mRNA levels of a selected set of inflammatory genes and promotes the activity of liver X receptor (LXR) and sterol regulatory element binding protein-1 (SREBP1). These acute antiinflammatory effects of 3'SL were associated with reduced histone H3K27 acetylation at a subset of LPS-inducible enhancers distinguished by preferential enrichment for CCCTC-binding factor (CTCF), IFN regulatory factor 2 (IRF2), B cell lymphoma 6 (BCL6), and other transcription factor recognition motifs. In a murine atherosclerosis model, both s.c. and oral administration of 3'SL significantly reduced atherosclerosis development and the associated inflammation. This study provides evidence that 3'SL attenuates inflammation by a transcriptional mechanism to reduce atherosclerosis development in the context of cardiovascular disease

Familial amyloid polyneuropathy: ocular complications and the use of novel non-invasive imaging techniques to assess retinal involvement

[eng] Familial amyloid polyneuropathy (FAP) is a hereditary condition characterized by systemic deposition of transthyretin (TTR), which causes debilitating peripheral polyneuropathy, cardiopathy, nephropathy and usually after a few years, ophtalmopathy. Occasionally the onset can be atypical and the diagnosis of FAP is reliant on identifying ocular amyloid deposition clinically and histopathologically. However, images of TTR derived from the eye, identified using immunolabeling techniques, have so far not been published. In ocular tissues FAP can cause sight-threatening complications such as glaucoma and retinal amyloid angiopathy. Glaucoma in FAP often requires surgical treatment. Nonpenetrating deep sclerectomy (NPDS) is a surgical technique with several advantages over the traditional trabeculectomy. It is successfully performed in primary and many types of secondary open-angle glaucoma, but so far with limited reports in FAP. Retina imaging modalities, such as optic coherence tomography (OCT) and autofluorescence (AF), have significant value in the assessment of retinal pathologies. Fluorescein angiography is the conventional method of evaluating retinal vasculature, but requires injection of fluorescein, which has several side effects and contraindications. Recently a new non-invasive modality, the OCT angiography (OCT-A) has become a useful tool for visualizing posterior pole blood circulation. In patients with FAP the use of OCT-A has so far not been reported and only few cases of AF findings were published. This doctoral thesis, presented as a compendium of publications, is divided into three parts. The first part (Paper 1) aims to present the immunostaining images of TTR amyloid derived from the vitreous of a series of patients with FAP, which demonstrates vitreous biopsy as a valid diagnostic tool, especially in clinically challenging cases. The second part (Paper 2) is a retrospective review of clinical charts of patients with FAP to determine the prevalence and characteristics of open-angle glaucoma secondary to FAP. It reveals the particularly quick progression of glaucoma in FAP and its increased risk in patients with a previous vitrectomy. Surgical management and outcomes of the affected patients are presented, indicating that NPDS is a safe and effective treatment of glaucoma secondary to FAP. The third part (Paper 3) is an observational cross-sectional study of retinal findings in patients with FAP. It gives a descriptive analysis of retinal images in FAP using novel non-invasive techniques: AF, OCT, OCT-A, and ultra-wide-field (UWF) retinography. These modalities can be used to detect perivascular retinal amyloid deposits, as well as microvascular changes including areas of non-perfusion, allowing better understanding of the pathology, complications and prognosis of patients with FAP. It also shows that amyloid retinopathy is more frequent than previously reported. The thesis outcomes emphasize glaucoma and retinopathy as the severe irreversible complications of FAP and need for addressing them promptly. This is especially important in establishing adequate regular eye reviews in patients with FAP and identifying those individuals requiring stricter ophthalmological care to prevent vision loss.[spa] La polineuropatía amiloidótica familiar (PAF) es una enfermedad hereditaria caracterizada por el depósito sistémico de transtiretina (TTR), que resulta en polineuropatía periférica debilitante, cardiopatía, nefropatía y, habitualmente, después de unos años, oftalmopatía. Ocasionalmente, el inicio puede ser atípico y el diagnóstico de PAF depende de la identificación de depósitos de amiloide en tejidos oculares clínicamente e histopatológicamente. Sin embargo, hasta ahora no se han publicado imágenes de TTR derivadas del ojo, identificadas utilizando técnicas de inmunotinción. En los tejidos oculares, la PAF puede causar complicaciones amenazantes para la vista, como el glaucoma y la angiopatía amiloide de la retina. El glaucoma en la PAF frecuentemente requiere tratamiento quirúrgico. La esclerectomía profunda no penetrante (EPNP) es una técnica quirúrgica con varias ventajas sobre la trabeculectomía tradicional. Se realiza con éxito en glaucoma de ángulo abierto primario y muchos tipos de glaucoma secundario, pero hasta ahora con pocos casos descritos en PAF. Las modalidades de imágen de retina, como la tomografía de coherencia óptica (OCT) y la autofluorescencia (AF), tienen un valor importante en la evaluación de las patologías retinianas. La angiografía con fluoresceína es el método convencional para evaluar la vasculatura retiniana, pero requiere la inyección de fluoresceína, que tiene varios efectos secundarios y contraindicaciones. Recientemente, una nueva modalidad no invasiva, la angiografía OCT (OCT-A) se ha convertido en una herramienta útil para visualizar la circulación sanguínea del polo posterior. En pacientes con PAF, el uso de OCT-A no ha sido publicado hasta ahora, y solo se han descrito dos casos de hallazgos de AF. Esta tesis doctoral, presentada como un compendio de publicaciones, se divide en tres partes. La primera parte (Artículo 1) tiene como objetivo presentar las imágenes de inmunotinción de TTR amiloide derivado del vítreo en una serie de pacientes con PAF, lo que demuestra que la biopsia vítrea es una herramienta de diagnóstico válida, especialmente en casos clínicamente atípicos. La segunda parte (Artículo 2) es una revisión retrospectiva de las historias clínicas de pacientes con PAF para determinar la prevalencia y las características del glaucoma de ángulo abierto secundario a la PAF. Revela la progresión particularmente rápida del glaucoma en la PAF y su mayor riesgo en pacientes con vitrectomía previa. Se ha presentado el tratamiento quirúrgico y los resultados de los pacientes afectados, lo que indica que EPNP es un tratamiento seguro y efectivo para el glaucoma secundario a PAF. La tercera parte (Artículo 3) es un estudio transversal observacional de hallazgos retinianos en pacientes con PAF. Se expone un análisis descriptivo de las imágenes de la retina en PAF utilizando nuevas técnicas no invasivas: AF, OCT, OCT-A y retinografía de campo amplio (UWF). Estas modalidades se pueden utilizar para detectar depósitos amiloides perivasculares de la retina, así como cambios microvasculares que incluyen áreas de no perfusión, lo que permite una mejor comprensión de la patología, las complicaciones y el pronóstico de los pacientes con PAF. También se muestra que la retinopatía amiloidea es más frecuente de lo que se publicó anteriormente. Los resultados de la tesis enfatizan el glaucoma y la retinopatía como las complicaciones irreversibles graves de la PAF y la necesidad de abordarlos precozmente. Esto es especialmente importante para establecer revisiones oculares regulares adecuadas en pacientes con PAF e identificar a aquellas personas que requieren atención oftalmológica más estricta para prevenir la pérdida de visión

Familial amyloid polyneuropathy: ocular complications and the use of novel non-invasive imaging techniques to assess retinal involvement

Familial amyloid polyneuropathy (FAP) is a hereditary condition characterized by systemic deposition of transthyretin (TTR), which causes debilitating peripheral polyneuropathy, cardiopathy, nephropathy and usually after a few years, ophtalmopathy. Occasionally the onset can be atypical and the diagnosis of FAP is reliant on identifying ocular amyloid deposition clinically and histopathologically. However, images of TTR derived from the eye, identified using immunolabeling techniques, have so far not been published. In ocular tissues FAP can cause sight-threatening complications such as glaucoma and retinal amyloid angiopathy. Glaucoma in FAP often requires surgical treatment. Nonpenetrating deep sclerectomy (NPDS) is a surgical technique with several advantages over the traditional trabeculectomy. It is successfully performed in primary and many types of secondary open-angle glaucoma, but so far with limited reports in FAP. Retina imaging modalities, such as optic coherence tomography (OCT) and autofluorescence (AF), have significant value in the assessment of retinal pathologies. Fluorescein angiography is the conventional method of evaluating retinal vasculature, but requires injection of fluorescein, which has several side effects and contraindications. Recently a new non-invasive modality, the OCT angiography (OCT-A) has become a useful tool for visualizing posterior pole blood circulation. In patients with FAP the use of OCT-A has so far not been reported and only few cases of AF findings were published. This doctoral thesis, presented as a compendium of publications, is divided into three parts. The first part (Paper 1) aims to present the immunostaining images of TTR amyloid derived from the vitreous of a series of patients with FAP, which demonstrates vitreous biopsy as a valid diagnostic tool, especially in clinically challenging cases. The second part (Paper 2) is a retrospective review of clinical charts of patients with FAP to determine the prevalence and characteristics of open-angle glaucoma secondary to FAP. It reveals the particularly quick progression of glaucoma in FAP and its increased risk in patients with a previous vitrectomy. Surgical management and outcomes of the affected patients are presented, indicating that NPDS is a safe and effective treatment of glaucoma secondary to FAP. The third part (Paper 3) is an observational cross-sectional study of retinal findings in patients with FAP. It gives a descriptive analysis of retinal images in FAP using novel non-invasive techniques: AF, OCT, OCT-A, and ultra-wide-field (UWF) retinography. These modalities can be used to detect perivascular retinal amyloid deposits, as well as microvascular changes including areas of non-perfusion, allowing better understanding of the pathology, complications and prognosis of patients with FAP. It also shows that amyloid retinopathy is more frequent than previously reported. The thesis outcomes emphasize glaucoma and retinopathy as the severe irreversible complications of FAP and need for addressing them promptly. This is especially important in establishing adequate regular eye reviews in patients with FAP and identifying those individuals requiring stricter ophthalmological care to prevent vision loss.La polineuropatía amiloidótica familiar (PAF) es una enfermedad hereditaria caracterizada por el depósito sistémico de transtiretina (TTR), que resulta en polineuropatía periférica debilitante, cardiopatía, nefropatía y, habitualmente, después de unos años, oftalmopatía. Ocasionalmente, el inicio puede ser atípico y el diagnóstico de PAF depende de la identificación de depósitos de amiloide en tejidos oculares clínicamente e histopatológicamente. Sin embargo, hasta ahora no se han publicado imágenes de TTR derivadas del ojo, identificadas utilizando técnicas de inmunotinción. En los tejidos oculares, la PAF puede causar complicaciones amenazantes para la vista, como el glaucoma y la angiopatía amiloide de la retina. El glaucoma en la PAF frecuentemente requiere tratamiento quirúrgico. La esclerectomía profunda no penetrante (EPNP) es una técnica quirúrgica con varias ventajas sobre la trabeculectomía tradicional. Se realiza con éxito en glaucoma de ángulo abierto primario y muchos tipos de glaucoma secundario, pero hasta ahora con pocos casos descritos en PAF. Las modalidades de imágen de retina, como la tomografía de coherencia óptica (OCT) y la autofluorescencia (AF), tienen un valor importante en la evaluación de las patologías retinianas. La angiografía con fluoresceína es el método convencional para evaluar la vasculatura retiniana, pero requiere la inyección de fluoresceína, que tiene varios efectos secundarios y contraindicaciones. Recientemente, una nueva modalidad no invasiva, la angiografía OCT (OCT-A) se ha convertido en una herramienta útil para visualizar la circulación sanguínea del polo posterior. En pacientes con PAF, el uso de OCT-A no ha sido publicado hasta ahora, y solo se han descrito dos casos de hallazgos de AF. Esta tesis doctoral, presentada como un compendio de publicaciones, se divide en tres partes. La primera parte (Artículo 1) tiene como objetivo presentar las imágenes de inmunotinción de TTR amiloide derivado del vítreo en una serie de pacientes con PAF, lo que demuestra que la biopsia vítrea es una herramienta de diagnóstico válida, especialmente en casos clínicamente atípicos. La segunda parte (Artículo 2) es una revisión retrospectiva de las historias clínicas de pacientes con PAF para determinar la prevalencia y las características del glaucoma de ángulo abierto secundario a la PAF. Revela la progresión particularmente rápida del glaucoma en la PAF y su mayor riesgo en pacientes con vitrectomía previa. Se ha presentado el tratamiento quirúrgico y los resultados de los pacientes afectados, lo que indica que EPNP es un tratamiento seguro y efectivo para el glaucoma secundario a PAF. La tercera parte (Artículo 3) es un estudio transversal observacional de hallazgos retinianos en pacientes con PAF. Se expone un análisis descriptivo de las imágenes de la retina en PAF utilizando nuevas técnicas no invasivas: AF, OCT, OCT-A y retinografía de campo amplio (UWF). Estas modalidades se pueden utilizar para detectar depósitos amiloides perivasculares de la retina, así como cambios microvasculares que incluyen áreas de no perfusión, lo que permite una mejor comprensión de la patología, las complicaciones y el pronóstico de los pacientes con PAF. También se muestra que la retinopatía amiloidea es más frecuente de lo que se publicó anteriormente. Los resultados de la tesis enfatizan el glaucoma y la retinopatía como las complicaciones irreversibles graves de la PAF y la necesidad de abordarlos precozmente. Esto es especialmente importante para establecer revisiones oculares regulares adecuadas en pacientes con PAF e identificar a aquellas personas que requieren atención oftalmológica más estricta para prevenir la pérdida de visión

<span style="font-size:11.0pt; mso-bidi-font-size:14.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN">Optimal<span style="font-size:11.0pt;mso-bidi-font-size: 14.0pt;font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US; mso-bidi-language:HI" lang="EN"> <span style="font-size: 11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN">concentration of<span style="font-size:11.0pt; mso-bidi-font-size:14.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-bidi-font-family:Mangal;mso-ansi-language:EN-GB; mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB"> hyaluronan<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN"> and<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Mangal; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB"> plant<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN"> protein<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Mangal; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB"> in different culture systems <span style="font-size:11.0pt; mso-bidi-font-size:14.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN">for<span style="font-size:11.0pt;mso-bidi-font-size: 14.0pt;font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-bidi-font-family:Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US; mso-bidi-language:HI" lang="EN"> <i style="mso-bidi-font-style: normal"><span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN">in vitro</span></i><span style="font-size:11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Mangal; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN"> <span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN">maturation<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Mangal; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN"> <span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN">of bovine<span style="font-size:11.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";mso-bidi-font-family:Mangal; mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN"> <span style="font-size:11.0pt;mso-bidi-font-size:14.0pt; font-family:"Times New Roman";mso-fareast-font-family:"Times New Roman"; mso-ansi-language:EN;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN">oocytes </span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span>

839-846With a view to search for<span style="mso-ansi-language: EN" lang="EN"> optimal<span style="mso-ansi-language: EN" lang="EN"> concentration of hyaluronan (HA) and plant protein (PP) in different culture systems for<span style="mso-ansi-language: EN" lang="EN"> in vitro maturation<span style="mso-ansi-language: EN" lang="EN"> of bovine<span style="mso-ansi-language: EN" lang="EN"> oocytes, cumulus-oocyte complexes (COCs) were matured in vitro in 2 culture systems (first co-cultured with granulose cells and estrus calf serum (ECS) in 2 mL volume, second without co-culture where ECS was replaced by exogenous hormones and BSA or PP in 100 µL dose under mineral oil). Seven types of media were used; 3 in first system and 4 in second system. To evaluate HA and PP effect on oocytes after in vitro culture an estimation of meiosis stage and a level of DNA fragmentation was performed by TUNEL staining. The highest meiotic maturation (84%) was observed in oocytes cultured in medium enriched with ECS in co-culture with granulose cells (1st system). The lowest meiotic maturation was noted in medium with addition of BSA (43%). The addition of HA in the medium enriched with BSA significantly increased the rate of matured oocytes (67%) and also didn’t affect the chromatin quality of individual oocytes. The addition of HA to the culture medium supplemented with a PP decreased the rate of matured oocytes to 54% but no statistical differences were noted. The results of the present study showed that HA supplementation didn’t have a detrimental impact on oocyte chromatin integrity and improved bovine oocytes’ meiotic maturation in medium supplemented only with BSA without co-culture of granulose cells. </span

Hydroxychloroquine retinopathy: an emerging problem.

Purpose The aim of this case series is to raise awareness of the emerging issue of serious retinal damage caused by the prolonged use of hydroxychloroquine (HCQ) and the importance of adequate and appropriate monitoring of visual function during treatment. Patients and methods This is a small retrospective case series of 3 patients on long-term HCQ who developed serious symptomatic retinal toxicity confirmed on imaging and functional testing. Results All 3 patients were treated with HCQ for over 15 years; two for rheumatoid arthritis (RA), and the third for systemic lupus erythematosus (SLE). All 3 patients had macular involvement varying in severity confirmed with characteristic features on imaging and functional testing (Optical Coherence Tomography (OCT), Autofluorescence (AF) and Humphrey 10-2 visual fields). Conclusion HCQ is widely used to treat autoimmune conditions with a proven survival benefit in patients with SLE. However, longterm use can be associated with irreversible retinal toxicity. These cases highlight that HCQ, like chloroquine, can also cause visual loss in susceptible individuals. Early detection of presymptomatic retinal changes by the introduction of appropriate screening and monitoring is mandatory to limit the extent of irreversible visual loss due to HCQ retinal toxicity.</p