Inês A. Amaral

Intervertebral disc (IVD) degeneration and herniation is a leading cause of disability globally and a large unmet clinical need. No efficient non-surgical therapy is available, and there is an urgency for minimally invasive therapies capable of restoring tissue function. IVD spontaneous hernia regression following conservative treatment is a clinically relevant phenomenon that has been linked to an inflammatory response. This study establishes the central role of macrophages in IVD spontaneous hernia regression and provides the first preclinical demonstration of a macrophage-based therapy for IVD herniation. A rat model of IVD herniation was used to test complementary experimental setups: (1) macrophage systemic depletion via intravenous administration of clodronate liposomes (Group CLP2w: depletion between 0 and 2 weeks post-lesion; Group CLP6w: depletion between 2 and 6 weeks post-lesion), and (2) administration of bone marrow-derived macrophages into the herniated IVD, 2 weeks post-lesion (Group Mac6w). Herniated animals without treatment were used as controls. The herniated area was quantified by histology in consecutive proteoglycan/collagen IVD sections at 2 and 6 weeks post-lesion. Clodronate-mediated macrophage systemic depletion was confirmed by flow cytometry and resulted in increased hernia sizes. Bone marrow-derived macrophages were successfully administered into rat IVD hernias resulting in a 44% decrease in hernia size. No relevant systemic immune reaction was identified by flow cytometry, cytokine, or proteomic analysis. Furthermore, a possible mechanism for macrophage-induced hernia regression and tissue repair was unveiled through IL4, IL17a, IL18, LIX, and RANTES increase. This study represents the first preclinical proof-of-concept of macrophage-based immunotherapy for IVD herniation.

Blueberry, an increasingly cultivated fruit crop in Portugal, is known to be susceptible to twig blight and dieback caused by species of Diaporthe. The diversity of Diaporthe species associated with symptomatic and asymptomatic Vaccinium corymbosum plants in Portugal was assessed. A multilocus sequence analysis of the rDNA internal transcribed spacer (ITS) region and the translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), calmodulin (cal), and histone 3 (his3) genes revealed the presence of Diaporthe foeniculina, Diaporthe rudis, and four new species, which are described as Diaporthe crousii, Diaporthe phillipsii, Diaporthe rossmaniae, and Diaporthe vacuae. These new species were characterized in terms of their morphology, mating strategies, and temperature growth requirements. In artificial inoculation trials of V. corymbosum cv. Bluecrop plants, all Diaporthe species caused minor symptoms. Further, no differences in aggressiveness were apparent between species. This study provides the first survey of Diaporthe species associated with blueberry twig blight and dieback in Portugal. It disclosed the occurrence of a diverse assemblage of Diaporthe species, whose status and impact as pathogens of blueberry is not yet fully understood.