Theoretical Accuracy of the Raytracing Method for Intraocular Calculation of Lens Power in Myopic Eyes after Small Incision Extraction of the Lenticule.

AIM: To evaluate the accuracy of the raytracing method for the calculation of intraocular lens (IOL) power in myopic eyes after small incision extraction of the lenticule (SMILE). METHODS: Retrospective study. All patients undergoing surgery for myopic SMILE between May 1, 2020, and December 31, 2020, with Scheimpflug tomography optical biometry were eligible for inclusion. Manifest refraction was performed before and 6 months after refractive surgery. One eye from each patient was included in the final analysis. A theoretical model was invited to predict the accuracy of multiple methods of lens power calculation by comparing the IOL-induced refractive error at the corneal plane (IOL-Dif) and the SMILE-induced change of spherical equivalent (SMILE-Dif) before and after SMILE surgery. The prediction error (PE) was calculated as the difference between SMILE-Dif-IOL-Dif. IOL power calculations were performed using raytracing (Olsen Raytracing, Pentacam AXL, software version 1.22r05, Wetzlar, Germany) and other formulae with historical data (Barrett True-K, Double-K SRK/T, Masket, Modified Masket) and without historical data (Barrett True-K no history, Haigis-L, Hill Potvin Shammas PM, Shammas-PL) for the same IOL power and model. In addition, subgroup analysis was performed in different anterior chamber depths, axial lengths, back-to-front corneal radius ratio, keratometry, lens thickness, and preoperative spherical equivalents. RESULTS: A total of 70 eyes of 70 patients were analyzed. The raytracing method had the smallest mean absolute PE (0.26 ± 0.24 D) and median absolute PE (0.16 D), and also had the largest percentage of eyes within a PE of ± 0.25 D (64.3%), ± 0.50 D (81.4%), ± 0.75 D (95.7%), and ± 1.00 D (100.0%). The raytracing method was significantly better than Double-K SRK/T, Haigis, Haigis-L, and Shammas-PL formulae in postoperative refraction prediction (all p < 0.001), but not better than the following formulae: Barrett True-K (p = 0.314), Barrett True-K no history (p = 0.163), Masket (p = 1.0), Modified Masket (p = 0.806), and Hill Potvin Shammas PM (p = 0.286). Subgroup analysis showed that refractive outcomes exhibited no statistically significant differences in the raytracing method (all p < 0.05). CONCLUSION: Raytracing was the most accurate method in predicting target refraction and had a good consistency in calculating IOL power for myopic eyes after SMILE.

Mouse macrophage polarity and ROCK1 activity depend on RhoA and non-apoptotic Caspase 3.

The macrophages have different subtypes with different functions in immune response and disease. It has been generally accepted that M1 macrophages are responsible for stimulation of immune system and inflammation while M2 macrophages play a role in tissue repair. Irrespective of the type, macrophage functions depend on actin cytoskeleton, which is under the control of small GTPase RhoA pathway and its downstream effector ROCK1. We generated RhoA-deleted macrophages and compared the effect of RhoA deletion on M0, M1 and M2 macrophage phenotype. Our studies showed that, unexpectedly, the RhoA deletion did not eliminate macrophage ROCK1 expression and increased ROCK1 activity. The RhoA deletion effect on macrophage phenotype, structure and polarity was different for each subtype. Moreover, our study indicates that the up-regulation of ROCK1 activity in RhoA-deleted macrophages and macrophage phenotype/polarity are dependent on non-apoptotic Caspase-3 and are sensitive to Caspase-3 inhibition. These novel findings will revise/complement our understanding of RhoA pathway regulation of cell structure and polarity.

ROCK inhibition impedes macrophage polarity and functions.

Macrophages play an important role in immune responses including allograft rejection and they are one of the potential targets of anti-rejection therapies in organ transplantation. Macrophage alloreactivity relies on their phenotype/polarity, motility, phagocytosis and matrix degradation, which in turn depend on proper functioning of actin cytoskeleton and its regulators, the small GTPase RhoA and its downstream effector the Rho-associated protein kinase (ROCK). Several laboratories showed that administration of ROCK inhibitor Y-27632 to the graft recipient inhibits chronic rejection or rodent cardiac allografts. Here we studied the effect of Y-27632 on mouse peritoneal macrophage structure, polarity and functions in in vitro assays. We show that Y-27632 inhibitor affects macrophage phenotype/polarity, phagocytosis, migration, and matrix degradation. These novel findings suggest that the impediment of macrophage structure and function via interference with the RhoA/ROCK pathway has a potential to be therapeutically effective in organ transplantation.

Dissonant response of M0/M2 and M1 bone-marrow-derived macrophages to RhoA pathway interference.

Macrophages have a multitude of functions in innate and adaptive immune response and organ and tissue homeostasis. Many experimental studies are performed on bone-marrow-derived macrophages differentiated in vitro into M1 (inflammatory) and M2 (anti-inflammatory) subtypes that express different molecular markers pertaining to their prospective functions. Macrophage phenotype, polarity and functions depend on the actin cytoskeleton, which is regulated by small GTPase RhoA, its downstream effector ROCK, and non-apoptotic Caspase-3. We generated transgenic mice with the macrophage-specific deletion of RhoA and compared the effect of Rho pathway interference (RhoA deletion and ROCK and Caspase-3 inhibition) on the phenotype, polarity and expression of subtype-specific molecular markers of bone-marrow-derived M0, M1 and M2 macrophages. We show that M0 and M2 macrophages have a radically different phenotype and polarity from M1 macrophages, and that this is mirrored in dissonant response to RhoA pathway interference. The RhoA pathway interference induces extreme elongation (hummingbird phenotype) of M0 and M2 but not M1 macrophages and inhibits the expression of M2-specific but not M1-specific molecular markers. These dramatic differences in the response of M0/M2 versus M1 macrophages to the same molecular cues ought to be important considerations in the interpretation of experimental data and therapeutic use of bone-marrow-derived macrophages.

Symmetry of upper eyelid after unilateral blepharoptosis repair with minimally invasive conjoint fascial sheath suspension technique.

AIM: To investigate the symmetry of upper eyelid in patients with unilateral mild and moderate blepharoptosis who underwent unilateral minimally invasive combined fascia sheath (CFS) suspension. METHODS: A retrospective study of patients who underwent unilateral minimally invasive CFS suspension surgery between January 2018 and December 2021. Inclusion criteria included unilateral mild and moderate ptosis, good levator muscle function (>9 mm) and follow-up of at least 6mo. Pre- and post-operative symmetry was graded subjectively for marginal reflex distance 1 (MRD1), tarsal platform show (TPS) and eyebrow fat span (BFS). A t-test was used to evaluate MRD1, TPS and BFS asymmetry by calculating delta values. The Bézier curve tool of the Image J software was used to extract the upper eyelid contours, where the symmetry was measured by the percentage of overlapping curvatures (POC). RESULTS: Totally 105 patients (105 eyelids) were included (mild group, n=84; moderate group, n=21). Postoperatively, all patients increased MRD1 and decreased TPS in the ptotic eye while maintaining unchanged BFS. The asymmetric delta value for MRD1 was measured to be 1.48±0.86 preoperatively, and it decreased to 0.58±0.67 postoperatively in all cases (P=0.0004). In patients with mild ptosis, the asymmetry value of TPS fell significantly from 1.15±0.62 to 0.68±0.38 (P=0.0187). The symmetry of the upper eyelid contour increased in all subgroups of patients, with a POC of 59.39%±13.45% preoperatively and POC of 78.29%±13.80% postoperatively. CONCLUSION: Minimally invasive CFS suspension is proved to be an effective means of improving the symmetry of unilateral ptosis in terms of MRD1 (all subgroups), POC (all subgroups) and TPS (only mild group), whereas BFS is unaffected.

Macrophages and RhoA Pathway in Transplanted Organs.

RhoA is a small GTPase that, via its downstream effectors, regulates a variety of cell functions such as cytokinesis, cell migration, vesicular trafficking, and phagocytosis. As such the RhoA pathway is also pivotal for proper functioning of immune cells including macrophages. By controlling actin cytoskeleton organization, RhoA pathway modulates macrophage's polarity and basic functions: phagocytosis, migration, and extracellular matrix degradation. Numerous studies indicate that macrophages are very important effectors contributing to acute and chronic rejection of transplanted organs. In this review we discuss the role of RhoA pathway in governance of macrophage's functions in terms of transplanted organs.

Macrophage/monocyte-specific deletion of Ras homolog gene family member A (RhoA) downregulates fractalkine receptor and inhibits chronic rejection of mouse cardiac allografts.

BACKGROUND: The cellular and molecular mechanisms of chronic rejection of transplanted organs remain obscure; however, macrophages are known to play a critical role in the injury and repair of allografts. Among multiple factors influencing macrophage infiltration to allografts, the fractalkine chemokine (C-X3-C motif) ligand 1(CX3CL1)/chemokine (C-X3-C motif) receptor 1 (CX3CR1) signaling pathway and actin cytoskeleton, which is regulated by a small guanosine-5׳-triphosphatase Ras homolog gene family member A (RhoA), are of the utmost importance. To define the role of macrophage/RhoA pathway involvement in chronic rejection, we generated mice with monocyte/macrophage-specific deletion of RhoA. METHODS: Hearts from BALB/c (H-2d) donors were transplanted into RhoAflox/flox (no Cre) and heterozygous Lyz2Cre+/-RhoAflox/flox recipients treated with cytotoxic T-lymphocyte-associated protein 4 immunoglobulin to inhibit early T-cell response. Allografts were assessed for chronic rejection and monocyte/macrophage functions. RESULTS: The deletion of RhoA inhibited macrophage infiltration, neointimal hyperplasia of vasculature, and abrogated chronic rejection of the allografts. The RhoA deletion downregulated G protein-coupled fractalkine receptor CX3CR1, which activates the RhoA pathway and controls monocyte/macrophage trafficking into the vascular endothelium. This in turn promotes, through overproliferation and differentiation of smooth muscle cells in the arterial walls, neointimal hyperplasia. CONCLUSIONS: Our finding of codependence of chronic rejection on monocyte/macrophage CX3CR1/CX3CL1 and RhoA signaling pathways may lead to the development of novel anti-chronic rejection therapies.

Macrophages as Effectors of Acute and Chronic Allograft Injury.

Organ transplants give a second chance of life to patients with end-stage organ failure. However, the immuno-logical barriers prove to be very challenging to overcome and graft rejection remains a major hurdle to long-term transplant survival. For decades, adaptive immunity has been the focus of studies, primarily based on the belief that T cells are necessary and sufficient for rejection. With better-developed immunosuppressive drugs and protocols that effectively control adaptive cells, innate immune cells have emerged as key effector cells in triggering graft injury and have therefore attracted much recent attention. In this review, we discuss current understanding of macrophages and their role in transplant rejection, their dynamics, distinct phenotypes, locations, and functions. We also discuss novel therapeutic approaches under development to target macrophages in transplant recipients.

Value of counting positive PHH3 cells in the diagnosis of uterine smooth muscle tumors.

The diagnosis of uterine smooth muscle tumors including leiomyosarcomas (LMS), smooth muscle tumors of uncertain malignant potential (STUMP), bizarre (atypical) leiomyoma (BLM), mitotically active leiomyoma (MAL) and leiomyoma (LM) depends on a combination of microscopic features, such as mitoses, cytologic atypia, and coagulative tumor cell necrosis. However, a small number of these tumors still pose difficult diagnostic challenges. The assessment of accurate mitotic figures (MF) is one of the major parameters in the proper classification of uterine smooth muscle tumors. This assessment can be hampered by the presence of increased number of apoptotic bodies or pyknotic nuclei, which frequently mimic mitoses. Phospho-histone H3 (PHH3) is a recently described immunomarker specific for cells undergoing mitoses. In our study, we collected 132 cases of uterine smooth muscle tumors, including 26 LMSs, 16 STUMPs, 30 BLMs, 30 MALs and 30 LMs. We used mitosis specific marker PHH3 to count mitotic indexes (MI) of uterine smooth muscle tumors and compared with the mitotic indexes of hematoxylin and eosin (H&E). There is a positive correlation with the number of mitotic figures in H&E-stained sections and PHH3-stained sections (r=0.944, P<0.05). The ratio of PHH3-MI to H&E-MI has no statistically significant difference in each group except for LMs (P>0.05). The counting value of PHH3 in LMSs have significantly higher than STUMPs, BLMs, MALs and LMs (P<0.001) and the counting value of PHH3 is 1.5±0.5 times of the number of mitotic indexes in H&E. To conclude, our results show that counting PHH3 is a useful index in the diagnosis of uterine smooth muscle tumors and it can provide a more accurate index instead of the time-honored mitotic figure counts at a certain ratio.