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The alternative pathway (AP) plays a major role in many complement-mediated human diseases. Factor D (FD), a rate-limiting enzyme in AP complement activation, is an attractive therapeutic target. Unlike other complement proteins, FD is synthesized primarily in adipose tissue, and its levels in human blood are relatively low. However, because of FD's high turnover rate, therapeutic targeting with monoclonal antibodies and chemical inhibitors has been challenging. The recent discovery that FD activity is regulated by mannose-binding lectin-associated serine protease 3 (MASP3), through conversion of a zymogen to mature FD, has sparked interest in MASP3 inhibition as a new way to block FD function and AP complement activity. Here, we review studies of mouse models of FD and MASP3 inhibition. We additionally discuss the lessons learned from these studies and their implications for therapeutic targeting of human FD and MASP3., (© 2024 The Author(s). European Journal of Immunology published by Wiley‐VCH GmbH.)

In the present research, novel lanthanide coordination compounds [DyL(PhCOO)(CH 3 OH)](ClO 4 ) 2 ·(CH 3 OH) 2 (1) were characterized by the compression of 2,6-diformyl-4-methyl-phenol (dmp) and 1,3-diamino-2-propanol using benzoate as the secondary ligand, where L indicates the deprotonated macrocyclic ligand. Through the high structural rigidity driven by the coordination of the macrocyclic ligand formed by condensation in methanol solution and sodium benzoate with Dy(ClO 4 ) 3 ·6H 2 O, compound 1 exhibits outstanding cyan-emitting fluorescence performance and potential applications as a fluorescent material. Additionally, hyaluronic acid (HA)/ carboxymethyl chitosan (CMCS) hydrogels were prepared with loaded resveratrol metal-organic complexes according to the synthetic chemical approach. In biological study, we evaluated the effect of hydrogels on oxidative stress on human dermal fibroblasts. Examined by molecular docking simulation, the results showed that the binding interactions were from the phenol group, the carboxyl group and also the "-N=" group, indicating Dy metal complex has excellent biological capability., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: The purpose of this study was to determine the distribution of the angular artery (AA) in the medial canthal area with the aim of defining an arterial course to prevent AA injury during facial surgery in this region., Methods: The authors dissected 36 hemifaces of 18 cadavers. The horizontal distance from the vertical level through the medial canthus to the AAs was measured. The AA course of each specimen was then recorded, and all of them were then superimposed to determine the AA course. The diameter and depth of the AA around the medial canthal area were also investigated using ultrasonography on living subjects., Results: The horizontal distances from the medial canthus level and 2 cm below the medial canthus were 9.0 ± 2.0 mm (mean ± SD) and 1.9 ± 2.4 mm, respectively. The superimposed image demonstrated that most of the AAs were present inside the vertical line through the medial canthus. Ultrasonography indicated that the AA was 2.3 ± 0.9 mm below the skin and 1.7 ± 0.3 mm in diameter., Conclusions: The AA course was relatively constant along the nasojugal fold. The AAs were most often present between the middle of the medial canthus and the facial midline, but were very scarce in both the medial and lateral thirds. Knowledge of the detailed course of the AA may help surgeons to avoid arterial injury and decrease the risk of surgical morbidities around the nasal root and medial canthal area., (Copyright © 2023 by the American Society of Plastic Surgeons.)

Thoracic aortic aneurysm/dissection (TAAD) is a lethal vascular disease, and several pathological factors participate in aortic medial degeneration. We previously discovered that the complement C3a-C3aR axis in smooth muscle cells promotes the development of thoracic aortic dissection (TAD) through regulation of matrix metalloproteinase 2. However, discerning the specific complement pathway that is activated and elucidating how inflammation of the aortic wall is initiated remain unknown. We ascertained that the plasma levels of C3a and C5a were significantly elevated in patients with TAD and that the levels of C3a, C4a, and C5a were higher in acute TAD than in chronic TAD. We also confirmed the activation of the complement in a TAD mouse model. Subsequently, knocking out Cfb (Cfb) or C4 in mice with TAD revealed that the alternative pathway and Cfb played a significant role in the TAD process. Activation of the alternative pathway led to generation of the anaphylatoxins C3a and C5a, and knocking out their receptors reduced the recruitment of inflammatory cells to the aortic wall. Moreover, we used serum from wild-type mice or recombinant mice Cfb as an exogenous source of Cfb to treat Cfb KO mice and observed that it exacerbated the onset and rupture of TAD. Finally, we knocked out Cfb in the FBN1 C1041G/+ Marfan-syndrome mice and showed that the occurrence of TAA was reduced. In summary, the alternative complement pathway promoted the development of TAAD by recruiting infiltrating inflammatory cells. Targeting the alternative pathway may thus constitute a strategy for preventing the development of TAAD. NEW & NOTEWORTHY The alternative complement pathway promoted the development of TAAD by recruiting infiltrating inflammatory cells. Targeting the alternative pathway may thus constitute a strategy for preventing the development of TAAD.

Complement constitutes a major part of the innate immune system. The study of complement in human health has historically focused on infection risks associated with complement protein deficiencies; however, recent interest in the field has focused on overactivation of complement as a cause of immune injury and the development of anticomplement therapies to treat human diseases. The kidneys are particularly sensitive to complement injury, and anticomplement therapies for several kidney diseases have been investigated. Overactivation of complement can result from loss-of-function mutations in complement regulators; gain-of-function mutations in key complement proteins such as C3 and factor B; or autoantibody production, infection, or tissue stresses, such as ischemia and reperfusion, that perturb the balance of complement activation and regulation. Here, we provide a high-level review of the status of anticomplement therapies, with an emphasis on the transition from rare diseases to more common kidney diseases.

Purpose: To evaluate the course of the cutaneous nerve regarding the first extensor compartment to determine whether the dorsal or volar approach is safer for local injection into the first extensor compartment guided by ultrasound., Methods: We dissected the radial side of the wrists from 28 cadavers (52 wrists). Four-points along the imaginary line were set: the styloid process and 1 cm, 2 cm, and 3 cm proximal to the styloid process. The numbers of superficial radial nerve (SRN) and lateral antebrachial cutaneous nerve (LACN) branches were counted, and distances from the imaginary line at these points and nerve diameters were recorded. Digital images were superimposed to observe overall distribution of cutaneous nerve., Results: There were means of 3.3 SRN and 0.9 LACN branches observed in each wrist. The mean number of both SRN and LACN branches was 2.3 on the dorsal side and 1.9 on the volar side. The superimposed images indicated that both the dorsal and volar sides comprised abundant cutaneous nerves and that their paths varied markedly between patients. However, we observed that larger nerves with meaningful diameters were more abundant on the dorsal than the volar side., Conclusion: There were similar numbers of cutaneous nerves on both the dorsal and volar sides; however, we observed greater abundance of thicker cutaneous nerves on the dorsal side, and these were closer to the reference line than on the volar side. This anatomical study suggests that the risk imposed to cutaneous nerves would therefore be reduced when injection on the volar side., Competing Interests: The authors declare no competing interests., (© 2024 The Authors.)

Seaweed extracts and their specific polysaccharides are widely known for their ability to act as reducing and capping agents during nanoparticle synthesis. Their application is highly favored in green synthesis methods, owing to their eco-friendliness, cost-effectiveness, and remarkable time and energy efficiency. In this study, fucoidan extracted from Undaria pinnatifida sporophyll (UPS) is introduced as a polysaccharide that effectively serves as a dual-function reducing and capping agent for the synthesis of gold nanoparticles (AuNPs). Results from various analyses indicate that AuNPs derived from UPS extract display a uniform spherical shape with an average size of 28.34 ± 1.15 nm and a zeta potential of -37.49 ± 2.13 mV, conclusively confirming the presence of Au. The FT-IR spectra distinctly revealed the characteristic fucoidan bands on the stabilized UPS-AuNPs surface. A 1 H-NMR analysis provided additional confirmation by revealing the presence of specific fucoidan protons on the UPS-AuNPs surface. To comprehensively evaluate the impact of UPS extract, UPS-AuNPs, and fucoidan on the biological properties of adipocytes, a rigorous comparative analysis of lipid droplet formation and morphology was conducted. Our findings revealed that adipocytes treated with UPS extract, fucoidan, and UPS-AuNPs, in that order, exhibited a reduction in the total lipid droplet surface area, maximum Ferret diameter, and overall Nile red staining intensity when compared to mature white adipocytes. Furthermore, our analysis of the effects of UPS extracts, UPS-AuNPs, and fucoidan on the expression of key markers associated with white adipose tissue browning, such as UCP1, PGC1a, and PRDM16, demonstrated increased mRNA and protein expression levels in the following order: UPS-AuNPs > fucoidan > UPS extracts. Notably, the production of active mitochondria, which play a crucial role in enhancing energy expenditure in beige adipocytes, also increased in the following order: UPS-AuNPs > fucoidan > UPS extract. These findings underscore the pivotal role of UPS extract, fucoidan, and UPS-AuNPs in promoting adipocyte browning and subsequently enhancing energy expenditure.

Background: The purpose of the current study was (1) to analyze various factors that may be associated with the outcomes of Legg-Calvé-Perthes disease (LCPD), and (2) to develop and internally validate machine learning algorithms capable of providing patient-specific predictions of which patients with LCPD will achieve relevant improvement in radiologic outcomes after proximal femoral varus osteotomy (PFVO). We examined several variables, previously identified as factors, that may influence the outcome of LCPD and developed a machine learning algorithm based on them., Methods: In this retrospective study, we analyzed patients aged older than  6 years at the time of LCPD diagnosis who underwent PFVO at our institution between 1979 and 2015. Univariate and multivariate logistic regression analyses were used to examine the effects of variables on the sphericity of the femoral head at skeletal maturity, including age at onset, sex, stage at operation, extent of epiphyseal involvement and collapse, presence of specific epiphyseal, metaphyseal, and acetabular changes, and postoperative neck shaft angle (NSA). Recursive feature selection was used to identify the combination of variables from an initial pool of 13 features that optimized the model performance. Five machine learning algorithms [extreme gradient boosting (XGBoost), multilayer perception, support vector machine, elastic-net penalized logistic regression, and random forest) were trained using 5-fold cross-validation 3 times and applied to an independent testing set of patients., Results: Ninety patients with LCPD who underwent PFVO were included in this study. The mean age at diagnosis was 7.93 (range, 6.0 to 12.33) years. The average follow-up period was 10.11 (range, 5.25 to 22.92) years. A combination of 8 variables, optimized algorithm performance, and specific cutoffs were found to decrease the likelihood of achieving the 1 or 2 Stulberg classification: age at onset ≤ 8.06, lateral classification ≤ B, 12.40 < preoperative migration percentage (MP) ≤ 22.85, Catterall classification ≤ 2, 117.4 < postoperative NSA ≤ 122.90, -10.8 < postoperative MP ≤ 6.5, 139.65 < preoperative NSA ≤ 144.67, and operation at stage 1. The XGBoost model demonstrated the best performance (F1 score: 0.78; area under the curve: 0.84)., Conclusions: The XGBoost machine learning algorithm achieved the best performance in predicting the postoperative radiologic outcomes in patients with LCPD who underwent PFVO. In our population, age at onset ≤ 8.06, lateral classification ≤ B, 12.40 < preoperative MP ≤ 22.85, Catterall classification ≤ 2, 117.4 < postoperative NSA ≤ 122.90, -10.8 < postoperative MP ≤ 6.5, 139.65 < preoperative NSA ≤ 144.67, and operation at an early stage had the likelihood of achieving the spherical femoral head for the patients with LCPD that underwent PFVO. After external validation, the online application of this model may enhance shared decision-making., Level of Evidence: Level III-retrospective cohort study., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Complement factor D (FD) is a rate-limiting enzyme of the alternative pathway (AP). Recent studies have suggested that it is synthesized as an inactive precursor and that its conversion to enzymatically active FD is catalyzed by mannan-binding lectin-associated serine protease 3 (MASP3). However, whether MASP3 is essential for AP complement activity remains uncertain. It has been shown that Masp1/3 gene knockout did not prevent AP complement overactivation in a factor H-knockout mouse, and a human patient lacking MASP3 still retained AP complement activity. In this study, we have assessed AP complement activity in a Masp3-knockout mouse generated by CRISPR/Cas9 editing of the Masp1/3 gene. We confirmed specific Masp3 gene inactivation by showing intact MASP1 protein expression and absence of mature FD in the mutant mice. Using several assays, including LPS- and zymosan-induced C3b deposition and rabbit RBC lysis tests, we detected plasma concentration-dependent AP complement activity in Masp3 gene-inactivated mice. Thus, although not measurable in 5% plasma, significant AP complement activity was detected in 20-50% plasma of Masp3 gene-inactivated mice. Furthermore, whereas FD gene deletion provided more than 90% protection of CD55/Crry-deficient RBCs from AP complement-mediated extravascular hemolysis, Masp3 gene deletion only provided 30% protection in the same study. We also found pro-FD to possess intrinsic catalytic activity, albeit at a much lower level than mature FD. Our data suggest that MASP3 deficiency reduces but does not abrogate AP complement activity and that this is explained by intrinsic pro-FD activity, which can be physiologically relevant in vivo., (Copyright © 2023 by The American Association of Immunologists, Inc.)

The M-type phospholipase A2 receptor (PLA2R) is the major autoantigen of primary membranous nephropathy (MN). Despite many studies on B-cell epitopes recognized by antibodies, little is known about T-cell epitopes. Herein, we synthesized 123 linear peptides, each consisting of 15-22 amino acids with 8-12 amino acid overlaps, across ten domains of PLA2R. Their binding capacity to risk (DRB1∗1501, DRB1∗0301) and protective (DRB1∗0901, DRB1∗0701) HLA molecules was then assessed by flow cytometry. Proliferation of CD4+ T cells from patients with anti-PLA2R positive MN was analyzed after peptide stimulation. Cytokines produced by activated peripheral blood mononuclear cells were measured by cytometric bead arrays. We identified 17 PLA2R peptides that bound to both DRB1∗1501 and DRB1∗0301 molecules with high capacity. Some of these peptides showed decreased binding to heterozygous DRB1∗1501/0901 and DRB1∗0301/0701. Ten of the 17 peptides (CysR1, CysR10, CysR12, FnII-3, CTLD3-9, CTLD3-10, CTLD3-11, CTLD5-2-1, CTLD7-1 and CTLD7-2) induced significant proliferation of CD4+ T cells from patients with MN than cells from healthy individuals. Upon activation by these peptides, peripheral blood mononuclear cells from patients with MN produced higher levels of pro-inflammatory cytokines, predominantly IL-6, TNF-α, IL-10, IL-9 and IL-17. Thus, we mapped and identified ten peptides in the CysR, FnII, CTLD3, CTLD5, and CTLD7 domains of PLA2R as potential T-cell epitopes of MN. These findings are a first step towards developing peptide-specific immunotherapies., (Copyright © 2022 International Society of Nephrology. All rights reserved.)

Despite their cytotoxic capacity, neutrophils are often co-opted by cancers to promote immunosuppression, tumor growth, and metastasis. Consequently, these cells have received little attention as potential cancer immunotherapeutic agents. Here, we demonstrate in mouse models that neutrophils can be harnessed to induce eradication of tumors and reduce metastatic seeding through the combined actions of tumor necrosis factor, CD40 agonist, and tumor-binding antibody. The same combination activates human neutrophils in vitro, enabling their lysis of human tumor cells. Mechanistically, this therapy induces rapid mobilization and tumor infiltration of neutrophils along with complement activation in tumors. Complement component C5a activates neutrophils to produce leukotriene B 4 , which stimulates reactive oxygen species production via xanthine oxidase, resulting in oxidative damage and T cell-independent clearance of multiple tumor types. These data establish neutrophils as potent anti-tumor immune mediators and define an inflammatory pathway that can be harnessed to drive neutrophil-mediated eradication of cancer., Competing Interests: Declaration of interests I.L.L., T.R.P., and E.G.E. are co-inventors on a patent application filed by Stanford University related to this work. W.-C.S. owns equity in, receives a consultant fee and research grant from, and is an inventor of patents licensed to Kira Pharmaceuticals and Aevitas Therapeutics. The other authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Heart surgery results in complement activation with the potential for collateral end-organ damage, especially if the protective elements (complement factor H, Apolipoprotein J) are inadequate. Here, we have investigated if peri-operative stress results in an imbalance between complement activation and its protective mechanisms up to 3 months after heart surgery., Methods: 101 patients scheduled for non-emergent cardiac surgery donated blood before the procedure (t baseline ), and 24 h (t 24 h ), 7 days (t 7 d ) and 3 months (t 3 m ) after. Complement activation was measured as a serum level of soluble activated component 5 (sC5a) and soluble terminal complement complex (sTCC). Simultaneously, protective complement factor H (CfH), and apolipoprotein J (ApoJ) were measured. Inflammatory responses were quantified using C-reactive protein (CRP) and interleukin-6 (IL-6). Details regarding anesthesia, intensive care unit (ICU) stay, pre-existing conditions, the incidence of postoperative complications, and mortality were collected from medical records., Results: C5a declined at t 24 h to rebound at t 7 d and t 3 m . sTCC was significantly depressed at t 24 h and returned to baseline at later time points. In contrast, CfH and ApoJ were depressed at t 3 m . Milieu of complement factors aligned along two longitudinal patterns:cluster#1 (C5a/sTTC continuously increasing and CfH/ApoJ preserved at t baseline ) and cluster#2 (transient sC5a/sTTC increase and progressive decline of CfH). Most patients belonged to cluster #1 at t 24 h (68%), t 7d (74%) and t 3m (72%). sTCC correlated with APACHE 1 h ( r 2 =-0.25; p < 0.031) and APACHE 24 h ( r 2 = 0.27; p < 0.049). IL-6 correlated with C5a ( r 2 =-0.28; p < 0.042) and sTTC ( r 2 =-0.28; p < 0.015). Peri-operative administration of acetaminophen and aspirin altered the complement elements. Prolonged hospital stay correlated with elevated C5a [ t (78) = 2.03; p = 0.048] and sTTC serum levels [ U (73) = 2.07; p = 0.037]. Patients with stroke had a decreased serum level of C5a at t 7d and t 3m ., Conclusion: There is a significant decrease in complement protective factors 3 months after cardiac surgery, while C5a seems to be slightly elevated, suggesting that cardiac surgery affects complement milieu long into recovery., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Laudanski, Liu, Gullipalli, Song, Okeke and Szeto.)

Background: The elevator muscles of the upper lip are the levator labii superioris alaeque nasi, levator labii superioris, and zygomaticus minor muscles, which function by means of their insertions into the skin of the upper lip. However, many textbooks and journal articles state that no muscle fibers are present on the orbicularis oris muscle in the upper lip. The authors attempted to determine whether there is a superficial muscle layer in addition to the orbicularis oris muscle in the upper lip., Methods: The authors performed gross dissections of 10 formalin-fixed cadavers and applied micro-computed tomography to six formalin-fixed cadavers. The fine dissection of the upper lip was performed in a layer-by-layer manner that elucidated its muscle layers. The entire layer of the upper lip was separated and pretreated with phosphotungstic acid for micro-computed tomography. The samples used for micro-computed tomography were repurposed for use in histologic analysis. An ultrasonography study was also performed., Results: The presence of a muscle layer on the orbicularis oris muscle was confirmed in all samples. The elevator muscle fibers of the upper lip formed a layer by combining with connective tissue. Micro-computed tomography indicated lower terminal insertions of the elevator muscles throughout the upper lip. All parts of the upper lip skin were inserted into the orbicularis oris muscle. The histologic findings were similar to those of micro-computed tomography., Conclusion: The authors' findings could be used to improve aesthetic and surgical procedures performed on the upper lip, such as correction of gummy smile and transverse upper labial crease, or postresection reconstruction of the upper lip., (Copyright © 2022 by the American Society of Plastic Surgeons.)

The facial artery is the main artery supplying blood to the face and is known to have facial branches of the inferior labial, superior labial, lateral nasal and angular arteries. These known major branches of facial artery run medially, however, there are sometimes branches of the facial artery heading laterally. The purpose of the present study was to investigate the lateral branches of the facial artery in face. We dissected facial branches of the facial artery in 74 cadaveric hemifaces. We investigated the presence of the lateral branches of the facial artery. Following parameters were investigated: lateral branch presence, the location of its origin, and the lateral branch diameter. Among the lateral branches, we evaluated the prevalence and diameter of the premasseteric branch. Lateral branches were observed in 48 of the 74 hemifaces (64.9%). The total number was 81 in the 48 hemifaces. The most common origin was between the inferior border of the mandible and inferior labial artery origin (42 of 81, 51.9%). The mean diameter of all lateral branches of the facial artery was 0.7 mm. Among the lateral branches, the premasseteric branches were present in 38 of 74 specimen (51.4%) and the mean diameter was 0.8 mm. The lateral branches of the facial artery may be registered in Terminologia Anatomica based on their prevalence. Accurate knowledge of the anatomy of the lateral branches of the facial artery is helpful for clinicians to avoid complications during facial procedures or maxillofacial surgeries., (© 2022 American Association of Clinical Anatomists and British Association of Clinical Anatomists.)

The purpose of this study is to identify the location of the orbital part of the lacrimal gland using external landmarks to facilitate effective botulinum toxin A (BTX-A) injections for epiphora treatment. Dissections were performed on 45 hemifaces from 27 cadavers. The length, anterior protrusion, and thickness of the orbital part of the lacrimal gland were measured directly. The midpoint of the line running horizontal through the medial and lateral canthus was used as the horizontal reference point. Vertical lines perpendicular to the midpoint of the horizontal reference line were the vertical reference lines. The angles from the horizontal reference line to the lacrimal gland center and to the frontal tubercle were also measured. The length and thickness of the lacrimal gland were 12.8 and 2.4 mm, respectively. The lacrimal gland had an anterior protrusion of 4.1 mm from the superolateral orbital margin. The superior and inferior margins of the lacrimal gland were located 35.7° and 15.8° from the two reference lines, respectively. The angle from the horizontal reference line to the lacrimal gland center was 35.1°, which was similar to that to the frontal tubercle (34.4°). The orbital part of the lacrimal gland was more inferior in elderly subjects and the angle between the horizontal reference line and its center was 35.1°, which was similar to the location of the frontal tubercle. The location of this tubercle can help clinicians to perform BTX-A injections into the lacrimal gland, thereby enhancing the effectiveness of epiphora treatment., (© 2022 American Association for Clinical Anatomists and the British Association for Clinical Anatomists.)

A complement effect on homeostasis during infection is determined by both cytotoxic (activate complement component 5 (C5a) terminal cytotoxic complex (TCC)), and cytoprotective elements (complement factor H (FH), as well as apolipoprotein E (ApoE)). Here, we investigated the gap in knowledge in their blood milieu during SARS-CoV-2 infection with respect to the viral burden, level of tissue necrosis, and immunological response. 101 patients hospitalized with a PCR-confirmed diagnosis of COVID-19 had blood collected at H1 (48 h), H2 (3-4 Days), H3 (5-7 days), H4 (more than 7 days up to 93 days). Pre-existing conditions, treatment, the incidence of cerebrovascular events (CVA), a history of deep venous thrombosis (DVT) and pulmonary embolism (PE), and mortality was collected using electronic medical records. Plasma C5a, TCC, FH, and ApoE were considered as a complement milieu. Tissue necrosis (HMGB1, RAGE), non-specific inflammatory responses (IL-6, C-reactive protein), overall viral burden (SARS-CoV-2 spike protein), and specific immune responses (IgG, IgA, IgM directed αS- & N-proteins) were assessed simultaneously. C5a remained elevated across all time points, with the peak at 5-7 days. Studied elements of complement coalesced around three clusters: #0 (↑↑↑C5a, ↑↑TCC, ↓↓ApoE), #1 ↑C5a, ↑TCC, ↑↑↑FH); #2 (↑C5a, ↑TCC, ↑FH, ↑↑↑ApoE). The decline in FH and ApoE was a predictor of death, while TCC and C5a correlated with patient length of stay, APACHE, and CRP. Increased levels of C5a (Δ = 122.64; p = 0.0294; data not shown) and diminished levels of FH (Δ = 836,969; p = 0.0285; data not shown) co-existed with CVA incidence. C5a correlated storngly with blood RAGE and HMGB1, but not with viral load and immunological responsiveness. Remdesivir positively affected FH preservation, while convalescent plasma treatment elevated C5a levels. Three clusters of complement activation demonstrated a various milieu of ApoE & FH vs C5a & TCC in COVID-19 patients. Complement activation is linked to increased necrosis markers but not to viral burden or immune system response., (© 2022. The Author(s).)

Laboratories use different strategies to sample and extract atmospheric particulate matter (PM), some of which can be very complicated. Due to the absence of a standard protocol, it is difficult to compare the results of PM toxicity assessment across different laboratories. Here, we proposed a novel PM sampling and cell exposure strategy based on agar membrane. The agar membrane, prepared by a simple freeze-drying method, has a relatively flat surface and porous interior. We demonstrated that the agar membrane was a reliable substitute material for PM sampling. Then the PM on the agar membranes was directly extracted with the culture medium by vortex method, and the PM on the polytetrafluoroethylene (PTFE) filters was extracted with water by the traditional ultrasonic method for comparison. The extraction efficiency was evaluated and in vitro cytotoxicity assays were carried out to investigate the toxic effects of PM extracted with two strategies on macrophage cells. The results showed that the PM extracted from agar membranes induced higher cytotoxicity and more differentially expressed proteins. Overall, the novel PM sampling-cell exposure strategy based on the agar membrane is easy to operate, biocompatible and comparable, and has low disturbance, could be an alternative sampling and extraction method for PM toxicity assessment., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The frontal sinus is one of the four paranasal sinuses in humans, and knowledge of its anatomy is important when performing surgery involving the frontal bone or sinus. Although many studies have measured the frontal sinus using radiography and computed tomography (CT), few studies have evaluated by using three-dimensional (3D) analysis. The purpose of this study was to analyze the frontal sinus using 3D reconstruction analysis and determine the differences in linear and volumetric measurements between sexes, sides, and ages. The sample comprised 281 facial CT scans: 173 and 108 from males and females, respectively. The width, height, and length of each frontal sinus and total volume were all larger in males than in females. Almost all linear and volumetric measurements were larger in young adults than in older for both sexes, but not all of the differences were statistically significant. Linear and volumetric measurements were larger for males than females regardless of age group. There were no statistically significant differences between the right and left sides except the width in males. The size of the frontal sinus was strongly influenced by sex and age. The measurements reported here might be useful for improving surgical procedures involving the frontal sinus.

Here, Au nanostructure (AuNS) biosynthesis was mediated through ethanolic extract of Plocamium telfairiae (PT) without the use of stabilizers or surfactants. PT-functionalized AuNSs (PT-AuNSs) were analyzed using ultraviolet-visible spectroscopy, dynamic light scattering, high-resolution transmission electron microscopy, energy-dispersive spectroscopy, and Fourier-transform infrared spectroscopy. Stable monodisperse PT-AuNSs were synthesized, with a mean size of 15.36 ± 0.10 nm and zeta potential of -35.85 ± 1.36 mV. Moreover, biosynthetic AuNPs with a face-centered structure of PT-AuNS exhibited crystalline characteristics. In addition, many functional groups playing important roles in the biological reduction of PT extracts were adsorbed on the surface of PT-AuNSs. Furthermore, the effects of PT-AuNSs on adipogenesis in immature adipocytes were investigated. PT-AuNSs reduced morphological changes, lowered triglyceride content, and increased lipid accumulation by approximately 78.6% in immature adipocytes compared with the values in mature adipocytes (MDI-induced). PT-AuNS suppressed lipid accumulation by downregulating the transcript and protein expression of C/EBPα, PPARγ, SREBP 1, FAS, and aP2. Finally, PT-AuNS induced the transcript and protein expression of UCP1, PRDM16, and PGC1a, thereby increasing mitochondrial biogenesis in mature adipocytes and effectively inducing brown adipogenesis. In this study, the biosynthesized PT-AuNS was used as a potential therapeutic candidate because it conferred a potent anti-lipogenic effect. As a result, it can be used in various scientific fields such as medicine and the environment.

Abstract: The zygomatico-orbital artery (ZOA) originating from the superficial temporal artery and supplying the lower temporal region superficially has been reported. Previous studies of this artery have used definitions that are too ambiguous for the results to be directly adapted to clinical practice, including since they have resulted in marked variations in the reported incidence ofthe artery. This study dissected 193 hemifaces of 123 fixed human cadavers aged 36 to 102 years (119 males and 74 females). The authors investigated the ZOA based on the following definition: (1) it originates from the superficial temporal artery, (2) it runs mostly above the zygomatic arch, and (3) it terminates below the superior border of the orbicularis oculi muscle. The incidence of the ZOA was 22.8% (44 cases of 193 sides), and its mean diameter was 1.1 mm. The meanvertical distances from the superior borderofthe zygomatic arch to the artery were 29.6, 17.8, and 2.9 mm at the jugale, zygion, and the origin of the ZOA, respectively. An accurate definition of the ZOA and accurate knowledge of its incidence and course could be important for clinicians to avoid unintentional complications in clinical practice., Competing Interests: The authors report no conflicts of interest., (Copyright © 2021 by Mutaz B. Habal, MD.)

A safe and effective vaccine against multidrug-resistant gonorrhea is urgently needed. An experimental peptide vaccine called TMCP2 that mimics an oligosaccharide epitope in gonococcal lipooligosaccharide, when adjuvanted with glucopyranosyl lipid adjuvant-stable emulsion, elicits bactericidal immunoglobulin G and hastens clearance of gonococci in the mouse vaginal colonization model. In this study, we show that efficacy of TMCP2 requires an intact terminal complement pathway, evidenced by loss of activity in C9-/- mice or when C7 function was blocked. In conclusion, TMCP2 vaccine efficacy in the mouse vagina requires membrane attack complex. Serum bactericidal activity may serve as a correlate of protection for TMCP2., (© The Author(s) 2021. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Background: The radial artery, a terminal branch of the brachial artery, typically originates at the level of the radial neck. It runs under the cover of the brachioradialis muscle, descending along the lateral side of the forearm with paired accompanying veins and a superficial branch of the radial nerve. The radial artery rewinds around the lateral aspect of the wrist, passing beneath the tendons of the abductor pollicis, extensor pollicis longus, and extensor pollicis brevis. Finally, it crossed the floor of the anatomical snuffbox., Case Report: This case report presents an anatomical variant in which the radial artery runs over the radial fossa, that is, superficially of the tendons of the hand's abductor pollicis, extensor pollicis longus, and extensor pollicis brevis muscles., Conclusion: The variation described in this paper sheds light on the possible atypical topographical relationships of the radial artery with the neighboring anatomical structures in the wrist, which can be crucial during hand surgery procedures., (© 2022. The Author(s), under exclusive licence to Springer-Verlag France SAS, part of Springer Nature.)

The rapid Cretaceous diversification of flowering plants remains Darwin's 'abominable mystery' despite numerous fossil flowers discovered in recent years. Wildfires were frequent in the Cretaceous and many such early flower fossils are represented by charcoalified fragments, lacking complete delicate structures and surface textures, making their similarity to living forms difficult to discern. Furthermore, scarcity of information about the ecology of early angiosperms makes it difficult to test hypotheses about the drivers of their diversification, including the role of fire in shaping flowering plant evolution. We report the discovery of two exquisitely preserved fossil flower species, one identical to the inflorescences of the extant crown-eudicot genus Phylica and the other recovered as a sister group to Phylica, both preserved as inclusions together with burned plant remains in Cretaceous amber from northern Myanmar (~99 million years ago). These specialized flower species, named Phylica piloburmensis sp. nov. and Eophylica priscastellata gen. et sp. nov., exhibit traits identical to those of modern taxa in fire-prone ecosystems such as the fynbos of South Africa, and provide evidence of fire adaptation in angiosperms., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Complement opsonization is among the biggest challenges facing nanomedicine. Nearly instantly after injection into blood, nanoparticles are opsonized by the complement protein C3, leading to clearance by phagocytes, fouling of targeting moieties, and release of anaphylatoxins. While surface polymers such as poly(ethylene glycol) (PEG) partially decrease complement opsonization, most nanoparticles still suffer from extensive complement opsonization, especially when linked to targeting moieties. To ameliorate the deleterious effects of complement, two of mammals' natural regulators of complement activation (RCAs), Factors H and I, are here conjugated to the surface of nanoparticles. In vitro, Factor H or I conjugation to PEG-coated nanoparticles decrease their C3 opsonization, and markedly reduce nanoparticle uptake by phagocytes. In an in vivo mouse model of sepsis-induced lung injury, Factor I conjugation abrogates nanoparticle uptake by intravascular phagocytes in the lungs, allowing the blood concentration of the nanoparticle to remain elevated much longer. For nanoparticles targeted to the lung's endothelium by conjugation to anti-ICAM antibodies, Factor I conjugation shifts the cell-type distribution away from phagocytes and toward endothelial cells. Finally, Factor I conjugation abrogates the severe anaphylactoid responses common to many nanoparticles, preventing systemic capillary leak and preserving blood flow to visceral organs and the brain. Thus, conjugation of RCAs, like Factor I, to nanoparticles is likely to help in nanomedicine's long battle against complement, improving several key parameters critical for clinical success., (© 2022 Wiley-VCH GmbH.)

Abstract: The present study is to identify primarily the morphological characteristics in the growth proportion of the head and face for young Korean (8-24 years) and compare the magnitude of growth changes to the sex-related differences. Total 1255 were divided into 3 age groups: childhood (8-10 years), adolescence (14-16 years), and young adult (20-24 years). The anthropometric assessments were performed with 11 landmarks on the head and facial dimensions. The standardized frontal and lateral head and face photographs were analyzed the craniofacial growth proportions and morphological features for the comparison of both sexes. The noteworthy differences of anthropometric measurements between sexes with growing were noted on the lower head height (22.6%, 17.8%), midface height (22.0%, 19.6%), lower face height (23.5%, 14.7%), and face length (21.1%, 14.9%), face breadth (14.8%, 11.3%) of males and females, respectively. Whereas the upper head height (7.9%, 6.0%) and upper face height (4.2%, 0%, respectively) were less growing features. The most remarkable changes are the dimension of midface height and lower face height in both sexes. The present study could demonstrate a fundamental example to elucidate the sex-related dimensional differences for the analysis of the growth proportion of both sexes in Koreans., Competing Interests: The authors report no conflicts of interest., (Copyright © 2021 by Mutaz B. Habal, MD.)

Feature screening is crucial in the analysis of ultrahigh dimensional data, where the number of variables (features) is in an exponential order of the number of observations. In various ultrahigh dimensional data, variables are naturally grouped, giving us a good rationale to develop a screening method using joint effect of multiple variables. In this article, we propose a group screening procedure via the F-test statistic. The proposed method is a direct extension of the original sure independence screening procedure, when the group information is known, for example, from prior knowledge. Under certain regularity conditions, we prove that the proposed group screening procedure possesses the sure screening property that selects all effective groups with a probability approaching one at an exponential rate. We use simulations to demonstrate the advantages of the proposed method and show its application in a genome-wide association study. We conclude that the grouping method is very useful in the analysis of ultrahigh dimensional data, as the optimal F-test can detect true signals with desired properties.

Nano-graphene oxide (Nano-GO) is an extensively studied multifunctional carbon nanomaterial with attractive applications in biomedicine and biotechnology. However, few studies have been conducted to assess the epithelial-to-mesenchymal transition (EMT) in the retinal pigment epithelium (RPE). We aimed to determine whether Nano-GO induces EMT by regulating phospholipase D (PLD) signaling in human RPE (ARPE-19) cells. The physicochemical characterization of Nano-GO was performed using a Zetasizer, X-ray diffraction, Fourier-transform infrared spectroscopy, and transmission electron microscopy. RPE cell viability assays were performed, and the migratory effects of RPE cells were evaluated. RPE cell collagen gel contraction was also determined. Intracellular reactive oxygen species (ROS) levels were determined by fluorescence microscopy and flow cytometry. Immunofluorescence staining and western blot analysis were used to detect EMT-related protein expression. Phospholipase D (PLD) enzymatic activities were also measured. Nano-GO significantly enhanced the scratch-healing ability of RPE cells, indicating that the RPE cell migration ability was increased. Following Nano-GO treatment, the RPE cell penetration of the chamber was significantly promoted, suggesting that the migratory ability was strengthened. We also observed collagen gel contraction and the generation of intracellular ROS in RPE cells. The results showed that Nano-GO induced collagen gel contraction and intracellular ROS production in RPE cells. Moreover, immunofluorescence staining and western blot analysis revealed that Nano-GO significantly regulated key molecules of EMT, including epithelial-cadherin, neural-cadherin, α-smooth muscle actin, vimentin, and matrix metalloproteinases (MMP-2 and MMP-9). Interestingly, Nano-GO-induced RPE cell migration and intracellular ROS production were abrogated in PLD-knockdown RPE cells, indicating that PLD activation played a crucial role in the Nano-GO-induced RPE EMT process. We demonstrate for the first time that Nano-GO promotes RPE cell migration through PLD-mediated ROS production. We provide preliminary evidence to support the hypothesis that Nano-GO has adverse health effects related to RPE damage.

COVID-19, the disease caused by the SARS-CoV-2 virus, can progress to multisystem organ failure and viral sepsis characterized by respiratory failure, arrhythmias, thromboembolic complications, and shock with high mortality. Autopsy and preclinical evidence implicate aberrant complement activation in endothelial injury and organ failure. Erythrocytes express complement receptors and are capable of binding immune complexes; therefore, we investigated complement activation in patients with COVID-19 using erythrocytes as a tool to diagnose complement activation. We discovered enhanced C3b and C4d deposition on erythrocytes in COVID-19 sepsis patients and non-COVID sepsis patients compared with healthy controls, supporting the role of complement in sepsis-associated organ injury. Our data suggest that erythrocytes may contribute to a precision medicine approach to sepsis and have diagnostic value in monitoring complement dysregulation in COVID-19-sepsis and non-COVID sepsis and identifying patients who may benefit from complement targeted therapies.

C3 glomerulopathy (C3G) is a rare renal disease characterized by predominant glomerular C3 staining. Complement alternative pathway dysregulation due to inherited complement defects is associated with C3G. To identify novel C3G-related genes, we screened 86 genes in the complement, coagulation and endothelial systems in 35 C3G patients by targeted genomic enrichment and massively parallel sequencing. Surprisingly, the most frequently mutated gene was VWF. Patients with VWF variants had significantly higher proteinuria levels, higher crescent formation and lower factor H (FH) levels. We further selected two VWF variants to transiently express the von Willebrand factor (vWF) protein, we found that vWF expression from the c.1519A > G variant was significantly reduced. In vitro results further indicated that vWF could regulate complement activation, as it could bind to FH and C3b, act as a cofactor for factor I-mediated cleavage of C3b. Thus, we speculated that vWF might be involved in the pathogenesis of C3G., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Volumetric rejuvenation of the dorsum of the hand with fat grafting or injectable fillers has increased in popularity in recent years. It has become widely accepted that the dorsum of the hand contains three fatty laminae: superficial, intermediate, and deep laminae. The dorsal venous plexus and dorsal cutaneous nerves are known to reside in the dorsal intermediate lamina. However, the superficial vein and cutaneous nerve might not be located in the same layer of subcutaneous tissue, as is the case in other body regions., Materials and Methods: Eight hands were dissected in a layer-by-layer fashion from the skin to the extensor tendons. In another 13 hands from among 21 investigated cadavers, samples from the dorsum of the hand were harvested and stained using trichrome stains for histologic analysis. B-mode ultrasound was also performed for identifying structures of the dorsum of the hand., Results: Anatomic dissection and histologic analysis of the dorsum of the hand revealed the presence of an unknown fascia in addition to the three known fascial layers. The additional fascia was located in the dorsal intermediate lamina and separated it into two compartments: one containing the dorsal venous plexus superficially and the other containing the dorsal cutaneous nerves deeply. Ultrasound showed corresponding structures including three hyperechoic fascial layers, three hypoechoic laminae, and additional hyperechoic fascia in dorsal intermediate lamina., Conclusions: Accurate anatomic knowledge of the dorsum of the hand will help practitioners determine the optimal and safe locations for performing fat grafting and injecting dermal fillers., (Copyright © 2020 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

In this study, the complete chloroplast (cp) genome of Duranta erecta was assembled using Illumina sequencing data. The complete cp genome is 149,869 bp in length, including a pair of invert repeats (IRA and IRB) regions of 22,839 bp, large single-copy (LSC) region of 86,201 bp, and small single-copy (SSC) region of 17,990 bp. The G + C content of this cp genome was 38.26%. A total of 128 genes were predicted in the genome, including 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis confirmed the phylogenetic relationship between D. erecta and other representative species of Verbenaceae., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Background: Retrobulbar filler injection has recently been considered an ideal method for orbital volume enhancement due to its nontoxic, easily reversible, and noninvasive characteristics. This study determined the arterial distribution in the orbit with the aim of defining a safety zone for retrobulbar filler injections used to enhance the orbital volume., Methods: Twenty-seven orbits of 24 formalin-embalmed cadavers were dissected. The orbital arteries were identified after removal of the eyeball, extraocular muscles, and connective tissues. The course of each orbital artery was then recorded in each specimen, and all of the courses were then superimposed to determine the arterial distribution in the orbit., Results: The superimposition of lined images based on the orbital vasculature of each specimen revealed that the arterial density was highest in the superonasal region and lowest in the inferotemporal region. In particular, orbital arteries were scarce at 8 o'clock and 4 o'clock in the right and left orbits, respectively, and an artery-free zone was demonstrated in the outer part of those directions., Conclusions: When performing a transcutaneous retrobulbar injection of filler for orbital volume enhancement, the relative safety zone could be considered to be located at 8 o'clock and 4 o'clock in the right and left orbits, respectively. The detailed topographic information about the arterial distribution in the orbit, provided by the present study, may help oculofacial surgeons to avoid injury to major vessels and decrease the risk of retrobulbar hemorrhage and vision-threatening complications., (Copyright © 2021 by the American Society of Plastic Surgeons.)

The complete chloroplast (cp) genome of Ehrharta erecta was sequenced and assembled for the first time. In this study, The total genome size is 134,511 bp in length and demonstrates a typical quadripartite structure containing a large single copy (LSC, 95,227 bp) and a small single copy (SSC, 12,306 bp), separated by a pair of inverted repeats (IRa, IRb) of 13,489 bp. The G + C content of this chloroplast genome was 38.76%. Gene annotation analysis identified 130 genes including 84 protein-coding genes, 38 transfer RNA, and 8 ribosomal RNA genes. The maximum-likelihood phylogenetic analysis result showed that E. erecta was closely related to O. sativa in the phylogenetic relationship., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

The statistical model (log-normal model), niche models (Zipf model, broken stick mo-del, niche preemption model), and neutral model were used to fit the species-abundance distribution patterns based on the measurements of environmental factors and inventory data of trees with DBH≥1 cm in a 1.5 hm 2 plot in the primary forest (PF) and a 1.5 hm 2 plot in the secondary forest (SF). The results showed that species-abundance distribution was affected by habitat heterogeneity in Q. aliena var . acutiserrata forest. Topography had a predominant impact on the species-abundance distribution in PF. Species distribution was affected by both neutral and niche processes, with neutral process having a less prominent effect in large convexity habitats. While the neutral model was rejected by the K-S and Chi-square test in low convexity habitats, the species-abundance distribution satisfied the assumption of niche theory. Niche process and neutral process were equally important in the community in areas with steep slopes, while niche differentiation was the dominant in flat areas. In SF, the main factors affecting species distribution were soil nutrients. The niche process was the mainly ecological process affected species-abundance distribution in habitats with high soil available phosphorus, while the niche and neutral processes existed simultaneously in habitats with low soil phosphorus availability. There was a significant scale effect on the species-abundance distribution pattern of Q. aliena var . acutiserrata forests in Taibai Mountain. The niche and neutral processes could protect the species-abundance distribution at the 20 m×20 m scale in PF, while the niche process could explain the species-abundance distribution at the 40 m×40 m and 70 m×70 m scales. The niche and neutral processes combined acted on the species abundance distribution at the 20 m×20 m, 40 m×40 m and 70 m×70 m scales in SF, with niche process being more important than neutral process. Moreover, besides the scale and habitat heterogeneity, the species-abundance distribution patterns of Q. aliena var. acutiserrata forests differed significantly between primary forest and secondary forest under anthropogenic disturbance.

In this study, the complete chloroplast genome of O. sativa Temperate Japonica YunJing-24 was assembled using Illumina sequencing data. The complete chloroplast (cp) genome is 134,556 bp in length, including a pair of invert repeats (IRA and IRB) regions of 20,797 bp, large single-copy (LSC) region of 80,615 bp, and small single-copy (SSC) region of 12,347 bp. A total of 129 genes were predicted in the genome, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis confirmed the phylogenetic relationship between O. sativa Temperate Japonica and other representative species., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2021 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

A layered coordination polymer (CP) with the fine-tuned alignment of four diolefinic ligands has been designed by shifting the coordination site of the ligand. The trimeric and tetrameric cyclobutane derivatives were reversely achieved by the photoinitiated [2+2] cycloaddition of the CP due to the favorable Schmidt's distance. More interestingly, a dynamic fluorescence shift was observed during the photo-oligomerization and heat-cycloreversion of the CP system.

The seminal vesicles are the glands of male reproductive organs that produce the fluid and nutrient constituents of semen. It has been believed for a long time that the lumen of a seminal vesicle was a single-coiled tubular structure with irregular diverticula. There are several previous reports on the symmetry, differences in morphological sizes and classification of the seminal vesicles. However, a three-dimensional-coiled tubular structure is difficult to understand using a classical anatomical methodology, and hence, three-dimensional reconstruction is needed to understand the structure of the lumen. Thirty-one seminal vesicles harvested from 21 formalin-embalmed cadavers were investigated. The seminal vesicle along with the ampulla of the ductus deferens was separated, and the length and width of each seminal vesicle were measured. The vesicles were then embedded in coloured paraffin, and the resulting paraffin block was sectioned transversely and photographed at an interval of 500 μm, with the sectioned surfaces then utilized in three-dimensional reconstruction performed by 'Reconstruct' software. The mean length and width of the seminal vesicles were 39.4 mm and 13.4 mm, respectively, and the right seminal vesicle was a little larger than the one on the left. The size differed from previous reports, while the luminal structure was similar to the classification of Aboul-azm (Archives of Andrology, 3, 1979, 287-292) but differed from that of Pereira (AJR. American Journal of Roentgenology, 69, 1953, 361-379). The seminal vesicles typically comprised about 9 curls and had about 12 diverticula. The seminal vesicles resembled a skein of coral rather than comprising a single strand. These findings will help in improving the understanding of pathophysiologies of the seminal vesicles, such as recurrent inflammation of the gland., (© 2020 Anatomical Society.)

Purpose: Knowledge of the distribution of intramuscular nerves of the extraocular muscles is crucial for understanding their function. The purpose of this study was to elucidate the intramuscular distribution of the oculomotor nerve within the inferior rectus muscle (IRM) using Sihler's staining., Method: Ninety-three IRM from 50 formalin-embalmed cadavers were investigated. The IRM including its branches of the oculomotor nerve was finely dissected from its origin to the point where it inserted into the sclera. The intramuscular nerve course was investigated after performing Sihler's whole-mount nerve staining technique that stains the nerves while rendering other soft tissues either translucent or transparent., Results: The oculomotor nerve enters the IRM around the distal one-fourth of the muscle and then divides into multiple smaller branches. The intramuscular nerve course finishes around the distal three-fifth of the IRM in gross observations. The types of branching patterns of the IRM could be divided into two subcategories based on whether or not topographic segregation was present: (1) no significant compartmental segregation (55.9% of cases) and (2) a several-zone pattern with possible segregation (44.1% of cases). Possible compartmentalization was less clear for the IRM, which contained overlapping mixed branches between different trunks., Conclusion: Sihler's staining is a useful technique for visualizing the gross nerve distribution of the IRM. The new information about the nerve distribution and morphological features provided by this study will improve the understanding of the biomechanics of the IRM, and could be useful for strabismus surgery.

Neuroblastoma (NB), which is a subtype of neural-crest-derived malignancy, is the most common extracranial solid tumor occurring in childhood. Despite extensive research, the underlying developmental origin of NB remains unclear. Using single-cell RNA sequencing, we generate transcriptomes of adrenal NB from 160,910 cells of 16 patients and transcriptomes of putative developmental cells of origin of NB from 12,103 cells of early human embryos and fetal adrenal glands at relatively late development stages. We find that most adrenal NB tumor cells transcriptionally mirror noradrenergic chromaffin cells. Malignant states also recapitulate the proliferation/differentiation status of chromaffin cells in the process of normal development. Our findings provide insight into developmental trajectories and cellular states underlying human initiation and progression of NB., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

This study aimed to elucidate the macroscopic and microscopic distributions of the dorsal nerve of penis (DNP) that provides the greatest sensitivity over the glans penis. The glandes of 23 penises of formalin-embalmed cadavers were investigated to confirm the macroscopic and microscopic distributions of the DNP within the glans penis by whole-mount Sihler's staining and histological sectioning. Superficial regions of the mid-glans were reconstructed in three dimensions to define the microstructure of terminal branches of the DNP that project towards the skin surface. A mean of 6.7 bundles of the DNP consisting of several nerve fibres converged linearly towards the distal end of the penis, rather than diverging laterally as they travelled. Lateral branches of the DNP extended linearly to the distal end with ramifications, while dorsomedial branches of the DNP gave off nerve fibres to the dorsum of the mid-glans and the corona. The intrastromal ramifications of the DNP were more developed in the distal half of the glans penis than the proximal glans containing the corpus cavernosum. These ramifications gave rise to radial nerve fibres that project towards the skin surface to form a plexiform network of terminal branches in the dermis. Linear projections of the main branches of the DNP throughout the glans and fine networks of terminal branches in the dermis were distinctly visualized in the human penis., (© 2020 Anatomical Society.)

Novel therapeutics against the global threat of multidrug-resistant Neisseria gonorrhoeae are urgently needed. Gonococci possess several mechanisms to evade killing by human complement, including binding of factor H (FH), a key inhibitor of the alternative pathway. FH comprises 20 short consensus repeat (SCR) domains organized in a head-to-tail manner as a single chain. N. gonorrhoeae binds two regions in FH; domains 6 and 7 and domains 18 through 20. We designed a novel anti-infective immunotherapeutic molecule that fuses domains 18-20 of FH containing a D-to-G mutation in domain 19 at position 1119 (called FH*) with human IgG1 Fc. FH*/Fc retained binding to gonococci but did not lyse human erythrocytes. Expression of FH*/Fc in tobacco plants was undertaken as an alternative, economical production platform. FH*/Fc was expressed in high yields in tobacco plants (300-600 mg/kg biomass). The activities of plant- and CHO-cell produced FH*/Fc against gonococci were similar in vitro and in the mouse vaginal colonization model of gonorrhea. The addition of flexible linkers [e.g., (GGGGS) 2 or (GGGGS) 3 ] between FH* and Fc improved the bactericidal efficacy of FH*/Fc 2.7-fold. The linkers also improved PMN-mediated opsonophagocytosis about 11-fold. FH*/Fc with linker also effectively reduced the duration and burden of colonization of two gonococcal strains tested in mice. FH*/Fc lost efficacy: i) in C6 -/- mice (no terminal complement) and ii) when Fc was mutated to abrogate complement activation, suggesting that an intact complement was necessary for FH*/Fc function in vivo . In summary, plant-produced FH*/Fc represent promising prophylactic or adjunctive immunotherapeutics against multidrug-resistant gonococci., (Copyright © 2020 Shaughnessy, Tran, Zheng, DeOliveira, Gulati, Song, Maclean, Wycoff and Ram.)

The iliacus muscle is a large, flat, triangle-shaped muscle located in the iliac fossa. This muscle forms part of the iliopsoas muscle complex. Although anatomical variations of iliacus muscles are rare, some variations are clinically important due to the possible coexistence of an unusual course of the femoral nerve. The femoral nerve is the largest branch of the lumbar plexus and supplies the muscles and skin in the anterior aspect of the thigh. We encountered a case of a single aberrant slip of the iliacus muscle piercing the femoral nerve in the left iliac fossa of a male cadaver aged 97 years. The potential clinical importance of this variant iliacus muscle accompanied by a femoral nerve split would be femoral neuropathy and possible consequent alterations of sensation in the anterior and medial aspects of the thigh or motor deficit of the quadriceps muscle.

Green-synthesized silver nanoparticles (SNPs) have great potential for biomedical applications, due to their distinctive optical, chemical, and catalytic properties. In this study, we aimed to develop green-synthesized SNPs from extracts of Cudrania tricuspidata (CT) roots (CTR), stems (CTS), leaves (CTL), and fruit (CTF) and to evaluate their physicochemical, photocatalytic, and biological properties. CTR, CTS, CTL, and CTF extracts were evaluated and compared for their total phenol and flavonoid content, reducing capacity, and antioxidant activity. The results revealed that CTR, CTS, CTL, and CTF extracts have high phenol and flavonoid content, as well as a powerful antioxidant and reducing capacity. CTR and CTS extracts showed the strongest effects. The results from UV-Vis spectra analysis, dynamic light scattering, high-resolution transmission electron microscopy, energy dispersive spectroscopy, X-ray diffraction, and Fourier-transform infrared spectroscopy showed the successful formation of CT-SNPs with surface morphology, crystallinity, reduction capacity, capsulation, and stabilization. Synthesized CT-SNPs successfully photocatalyzed methylene blue, methyl orange, rhodamine B, and Reactive Black 5 within 20 min. The CTR- and CTS-SNPs showed better antibacterial properties against different pathogenic microbes ( Staphylococcus aureus , Bacillus cereus , Escherichia coli , and Salmonella enteritidis ) than the CTL- and CTF-SNPs. CTS- and CTR-SNPs showed the most effective cytotoxicity and antiapoptosis properties in human hepatocellular carcinoma cells (HepG2 and SK-Hep-1). CT-SNPs also seemed to be more biologically active than the CT extracts. The results of this study provide evidence of the establishment of CT extract SNPs and their physicochemical, photocatalytic, and biological properties.

Osteoporosis is a major disease in aged women, increasing the risk for fractures accompanied by changes in the microarchitecture. The aim of this study was to investigate the three-dimensional (3D) histomorphology of femur diaphysis in the animal model for postmenopausal osteoporosis. The cortical bone of femur diaphysis of the rat was serially sectioned at a thickness of 5 μm and evaluated age-associated changes of the intracortical (osteonal) canal networks three-dimensionally. Cortical microstructures of 10-month old rats were not affected by ovariectomy. Intracortical canal networks were radial toward endosteal aspect and frequently interconnected across the neighboring canals with short arciform and irregular canals reminiscent for resorption spaces in ovarectomized 16-month old rats, contrary to intact canals in 16-month old control rat. Increased proportion of the periosteal circumference lamella and deformed endosteal regions with rare cortical canals hampered reconstructive histomorphology in ovarectomized rats of 26 month age. We have shown that 3D reconstruction of rat femur of the aged model over 16-month old is suitable methods that evaluate and microstructural change of the intracortical canals and cortical bone porosity by estrogen depletion.

Background: The deep temporal fascia (DTF) is known to separate into two layers that descend to attach to the zygomatic arch. When surgeons reduce an isolated fracture of the zygomatic arch through a temporal approach, the temporal incision site needs to be superior to the split line of the DTF., Materials and Methods: Sixty-seven hemifacial cadavers were investigated after removing the skin, subcutaneous tissue, and superficial temporal fascia. The superficial layer of the DTF was exposed. We cut the superficial layer along the line along, which it adhered to the deep layer inseparably. The heights of the split line of the DTF from the superior border of the zygomatic arch and from the top of the helix were measured at three points: at the jugale, zygion, and 3 cm from the tragus., Results: In all cases there were thick identifiable deep layers of the DTF. The mean heights of the split line of the DTF from the superior border of the zygomatic arch were 49.8, 46.7, and 42.6 mm at the jugale, zygion, and 3 cm from the tragus, respectively; the corresponding mean heights of the split line from the top of the helix were 19.1, 15.6, and 11.4 mm., Conclusions: Knowledge of the mean height of the split line of the DTF will be helpful for surgeons to determine the temporal incision site for ensuring the safe reduction of a zygomatic arch fracture., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest concerning the materials or methods used in this study and no source of funding., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Yu Gan Long (YGL) is a Chinese traditional herbal formula which has been reported to attenuate liver fibrosis for many years and we have explored its anti-fibrotic mechanism through blocking transforming growth factor (TGF-β) in the previous study. But the mechanisms associated with platelet-derived growth factor (PDGF)-BB remain obscure. In this study, we further investigated the mechanism of YGL reducing carbon tetrachloride (CCl 4 )-induced liver fibrosis in rats. Our results showed that YGL suppressed CCl 4 -induced upregulation of collagen IV (Col IV), type HI precollagen (PCHI), hyaluronuc acid (HA) and laminin (LN), which are implicated in liver fibrosis. Also, YGL reduced the α-smooth muscle actin (α-SMA) expression, which acts as the indicator of liver fibrosis. Furthermore, YGL decreased the serum levels of hepatic stellate cell (HSC) mitogen PDGF-BB and inflammation cytokines, including TNF-α, IL-1β, IL-6. Markers involved in liver fibrosis, such as Ras, p-Raf-1, p-ERK1/2, p-JNK, p-P38, p-PI3K, p-AKT, p-JAKl, p-STAT3 were downregulated significantly after treatment with YGL. Our results indicated that YGL ameliorated CCl 4 -induced liver fibrosis by reducing inflammation cytokines production, and suppressing Ras/ERK, PI3K/AKT, and JAK1/STAT3 signaling pathways, which provided further evidence towards elucidation of the anti-fibrotic mechanism of YGL.

Frankfort horizontal line, the line passing through the orbitale and porion, is one of the most widely used intracranial landmarks in cephalometric analysis. This study investigated the use of the orbito-occipital line extending from the orbitale to the external occipital protuberance as a novel horizontal line of the skull for substituting the Frankfort horizontal line. We evaluated the reproducibility of the new landmark and measured the angle between the orbito-occipital line and the Frankfort line. This study was conducted on 170 facial computed tomography (CT) scans of living adults from the Department of Plastic Surgery. After three-dimensionally reconstructed images were obtained from facial CT, the porion, orbitale, and external occipital protuberance were indicated by two observers twice. The angles between the orbito-meatal line (inferior orbital rim to porion; the Frankfort line) and the orbito-occipital line (inferior orbital rim to external occipital protuberance) were measured. There was no significant intraobserver or interobserver bias. The overall angle between the Frankfort line and orbito-occipital line was -0.5°±2.2° (mean±standard deviation). There was no statistically significant difference among side and sex. This study demonstrated good reproducibility of a new landmark-the external occipital protuberance-tested to replace the porion. The orbito-occipital line is a reliable, reproducible, and easily identifiable line, and has potential as a novel standard horizontal line to replace or at least supplement the Frankfort line in anthropological studies and certain clinical applications., Competing Interests: Conflicts of Interest: No potential conflict of interest relevant to this article was reported., (Copyright © 2020. Anatomy & Cell Biology.)