[IS ROUTINE CT SCAN NECESSARY AFTER THE REMOVAL OF BENIGN BRAIN TUMORS?]

AIMS: Determine if early CT evaluation is justified in patients operated on for benign brain tumors. BACKGROUND: Researchers have recently questioned the common practice of referring all patients after cranial surgery for CT scans to rule out silent complications. METHODS: The cohort included 257 consecutive patients who underwent surgical removal of a benign brain tumor in the years 2011-2016. The neuroradiology scans performed before and after surgery were reviewed. The postoperative findings considered significant were hemorrhage in ≥50% of the tumor bed, ischemic changes, worsening brain edema, and mass effect. The relationship of the CT findings with the neurological outcome and their effect on the clinical management were evaluated. RESULTS: No significant complications were found by CT in 247 patients (96%). In the remaining 10 patients (4%), CT showed increased brain edema in 5 and hemorrhage in 5. The clinical management was influenced by the CT findings in 6/10 patients. One had a temporary neurological deficit. None died or required a second operation. CONCLUSIONS: Elective patients operated on uneventfully for benign brain tumors rarely benefit from routine CT after surgery. In most cases, CT follow-up can be replaced by careful neurological monitoring. Exceptions to this rule may be based on clinical judgment and local resources.

Integrin restriction by miR-34 protects germline progenitors from cell death during aging.

During aging, regenerative tissues must dynamically balance the two opposing processes of proliferation and cell death. While many microRNAs are differentially expressed during aging, their roles as dynamic regulators of tissue regeneration have yet to be described. We show that in the highly regenerative Drosophila testis, miR-34 levels are significantly elevated during aging. miR-34 modulates germ cell death and protects the progenitor germ cells from accelerated aging. However, miR-34 is not expressed in the progenitors themselves but rather in neighboring cyst cells that kill the progenitors. Transcriptomics followed by functional analysis revealed that during aging, miR-34 modifies integrin signaling by limiting the levels of the heterodimeric integrin receptor αPS2 and ßPS subunits. In addition, we found that in cyst cells, this heterodimer is essential for inducing phagoptosis and degradation of the progenitor germ cells. Together, these data suggest that the miR-34-integrin signaling axis acts as a sensor of progenitor germ cell death to extend progenitor functionality during aging.

Sucrose Synthase and Fructokinase Are Required for Proper Meristematic and Vascular Development.

Sucrose synthase (SuSy) and fructokinase (FRK) work together to control carbohydrate flux in sink tissues. SuSy cleaves sucrose into fructose and UDP-glucose; whereas FRK phosphorylates fructose. Previous results have shown that suppression of the SUS1,3&4 genes by SUS-RNAi alters auxin transport in the shoot apical meristems of tomato plants and affects cotyledons and leaf structure; whereas antisense suppression of FRK2 affects vascular development. To explore the joint developmental roles of SuSy and FRK, we crossed SUS-RNAi plants with FRK2-antisense plants to create double-mutant plants. The double-mutant plants exhibited novel phenotypes that were absent from the parent lines. About a third of the plants showed arrested shoot apical meristem around the transition to flowering and developed ectopic meristems. Use of the auxin reporter DR5::VENUS revealed a significantly reduced auxin response in the shoot apical meristems of the double-mutant, indicating that auxin levels were low. Altered inflorescence phyllotaxis and significant disorientation of vascular tissues were also observed. In addition, the fruits and the seeds of the double-mutant plants were very small and the seeds had very low germination rates. These results show that SUS1,3&4 and FRK2 enzymes are jointly essential for proper meristematic and vascular development, and for fruit and seed development.

The phagocytic cyst cells in Drosophila testis eliminate germ cell progenitors via phagoptosis.

Phagoptosis is a frequently occurring nonautonomous cell death pathway in which phagocytes eliminate viable cells. While it is thought that phosphatidylserine (PS) "eat-me" signals on target cells initiate the process, the precise sequence of events is largely unknown. Here, we show that in Drosophila testes, progenitor germ cells are spontaneously removed by neighboring cyst cells through phagoptosis. Using live imaging with multiple markers, we demonstrate that cyst cell-derived early/late endosomes and lysosomes fused around live progenitors to acidify them, before DNA fragmentation and substantial PS exposure on the germ cell surface. Furthermore, the phagocytic receptor Draper is expressed on cyst cell membranes and is necessary for phagoptosis. Significantly, germ cell death is blocked by knockdown of either the endosomal component Rab5 or the lysosomal associated protein Lamp1, within the cyst cells. These data ascribe an active role for phagocytic cyst cells in removal of live germ cell progenitors.

CD200 -dependent and -independent immune-modulatory functions of neural stem cells.

Neural stem/precursor cells (NPC) exhibit powerful immune-modulatory properties. Attenuation of neuroinflammation by intra-cerebroventricular transplantation of NPC, protects from immune-mediated demyelination and axonal injury. The immune modulatory properties of NPC are mediated by a non-species-specific, multiple bystander effect, mediated by both direct cell-cell contact, and by soluble factor(s). CD200 is a cell-surface molecule, with important roles in regulating diverse immune responses, and shown also to limit neuroinflammatory processes. We hypothesized that CD200 may play a role in mediating immune-modulatory effects of NPC. We used wild type and CD200-deficient NPC to examine the role of CD200 in mediating two vital aspects of NPC -immune modulatory properties: (1) Attenuation of autoimmune neuroinflammation; and (2) Suppression of immune rejection response towards transplanted allogeneic NPC from the host CNS. We found that CD200 is dispensable for attenuating acute experimental autoimmune neuroinflammation, but is required for protecting transplanted allogeneic NPC from immune rejection by the host tissue. CD200 deficient NPC showed similar growth, differentiation and survival properties as wild type NPC. CD200-deficient NPC attenuated efficiently T cell activation and proliferation, but exhibited reduced ability to inhibit macrophages. We conclude that CD200 plays a partial role in mediating the immune-modulatory properties of NPC. The differential effect on T cells versus macrophages may underlie the observed discrepancy in their function in vivo.

Expression of Hexokinase in Stomata of Citrus Fruit Reduces Fruit Transpiration and Affects Seed Development.

The temporal formation and spatial distribution of stomata on the surface of citrus floral organs and, specifically, on the ovule from which the fruit develops, were analyzed using citrus plants that express green fluorescent protein (GFP) under the guard cell-specific KST1 promoter. Stomata are found on the style, sepal, and anther of the closed flower and on ovules from the stage of anthesis. It has previously been shown that hexokinase (HXK) mediates sugar-sensing in leaf guard cells and stimulates stomatal closure. The activity and response of citrus fruit stomata to sugar-sensing by HXK was examined using plants that express HXK under the KST1 promoter. Those plants are referred to as GCHXK plants. The transpiration of young green GCHXK citrus fruits was significantly reduced, indicating that their stomata respond to sugar similar to leaf stomata. Toward fruit maturation, fruit stomata are plugged and stop functioning, which explains why WT and GCHXK mature yellow fruits exhibited similar water loss. Seeds of the GCHXK plants were smaller and germinated more slowly than the WT seeds. We suggest that the stomata of young green citrus fruits, but not mature yellow fruits, respond to sugar levels via HXK and that fruit stomata are important for proper seed development.

Expression and characterization of the thylakoid lumen protease DegP1 from Arabidopsis.

The Arabidopsis genome contains 14 genes encoding the serine protease DegP. Products of four of these genes are located in the chloroplast: three in the thylakoid lumen and one on the stromal side of the membrane. We expressed the gene encoding DegP1 as a His-tagged fusion protein in Escherichia coli, purified the protein by affinity chromatography, and characterized it biochemically. Size-exclusion chromatography suggested that DegP1 eluted from the column as a mixture of monomers and hexamers. Proteolytic activity was characterized using beta-casein as a model substrate. DegP1 demonstrated concentration-dependent activity, a pH optimum of 6.0 and increasing activity at elevated temperatures. DegP1 was capable of degrading two lumenal proteins, plastocyanin and OE33, suggesting a role as a general-purpose protease in the thylakoid lumen. The results of this work are discussed in the context of the recent elucidation of the structure of the E. coli homolog and the possible physiological role of the protease in the chloroplast lumen.