Serum soluble CD163 and proinflammatory chemokines may be biomarkers of the onset of adverse events in dabrafenib plus trametinib combination therapy for advanced melanoma

Various adverse events (AEs) have been reported to occur at a high rate in patients treated with dabrafenib plus trametinib (D + T) combination therapy. Among such AEs, the incidence of pyrexia was highest among the series of AEs in patients treated with D + T combination therapy. Although little is known about the mechanisms of pyrexia caused by D + T combination therapy, a recent report suggested that sCD163, as well as interferon‐inducible chemokines (CXCL9, CXCL10, CXCL11), might correlate with pyrexia caused by encorafenib plus binimetinib combination therapy. In addition to these soluble factors, CXCL5 is a biomarker for predicting immune‐related AEs in melanoma patients treated with nivolumab. From the above findings, we hypothesized that these soluble factors might also correlate with the onset of AEs in D + T combination therapy. The serum levels of sCD163 were increased in patients with pyrexia in parallel with their severity, whereas the serum levels of CXCL5 were increased in patients without pyrexia. Moreover, increased levels of CXCL9, CXCL10, and CXCL11 were prominent in patients with AEs over G2 levels. As these chemokines recruit Th1, Th17, and activated CD8+ T cells, increased serum levels of these chemokines might correlate with the positive feedback of inflammatory reactions related to AEs.     

Awaji ALS criteria increase the diagnostic sensitivity in patients with bulbar onset

ObjectiveTo assess whether Awaji criteria improve the sensitivity of diagnosis for amyotrophic lateral sclerosis (ALS). In Awaji ALS criteria, fasciculation potentials are regarded as evidence of acute denervation in the presence of chronic neurogenic changes on needle electromyography.MethodsWe reviewed clinical and neurophysiological data of 113 consecutive patients who were suspected as suffering ALS. The six muscles (trapezius, biceps, first dorsal interosseous, T10-paraspinalis, vastus lateralis, and tibialis anterior muscles) were examined by EMG, focusing on the presence of fasciculation potentials. The sensitivity of revised El Escorial (R-EEC) and Awaji criteria was compared.ResultsProbable or definite ALS was diagnosed in 61% of the patients by R-EEC and 71% by Awaji criteria. By applying Awaji criteria; (1) 17 of the 44 patients categorized as possible ALS by R-EEC reached to probable/definite ALS, 11 of whom had bulbar onset, (2) in 48 patients with bulbar onset, the proportion of probable/definite ALS increased from 59% to 82%, (3) in 62 patients with limb onset, the proportion of probable/definite ALS was 61% (63% by R-EEC).ConclusionsAwaji criteria improve the sensitivity of ALS diagnosis in patients with bulbar onset, but not in those with limb onset.SignificanceAccepting fasciculation potentials as evidence of acute denervation increases the diagnostic sensitivity of ALS, particularly in patients with bulbar onset, and contributes to early diagnosis.     

Robust designation of meiotic crossover sites by CDK-2 through phosphorylation of the MutSγ complex

Crossover formation is essential for proper segregation of homologous chromosomes during meiosis. Here, we show that Caenorhabditis elegans cyclin-dependent kinase 2 (CDK-2) partners with cyclin-like protein COSA-1 to promote crossover formation by promoting conversion of meiotic double-strand breaks into crossover–specific recombination intermediates. Further, we identify MutSγ component MSH-5 as a CDK-2 phosphorylation target. MSH-5 has a disordered C-terminal tail that contains 13 potential CDK phosphosites and is required to concentrate crossover–promoting proteins at recombination sites. Phosphorylation of the MSH-5 tail appears dispensable in a wild-type background, but when MutSγ activity is partially compromised, crossover formation and retention of COSA-1 at recombination sites are exquisitely sensitive to phosphosite loss. Our data support a model in which robustness of crossover designation reflects a positive feedback mechanism involving CDK-2–mediated phosphorylation and scaffold-like properties of the MSH5 C-terminal tail, features that combine to promote full recruitment and activity of crossover–promoting complexes.

Sexually reproducing organisms rely on proper chromosome segregation during meiosis to produce gametes with a complete genome. During meiotic prophase I, chromosomes pair and undergo crossover recombination with their homologous partners. This process, together with sister chromatid cohesion, leads to the formation of physical linkages between the homologs and enables their separation during meiosis I. Defects in crossover formation are disastrous, leading to miscarriages and congenital disorders, such as Down syndrome (1).Meiotic recombination initiates with the generation of programmed DNA double-strand breaks (DSBs) by the topoisomerase-like enzyme Spo11 (2). DSBs are resected to yield two 3′-end single-stranded DNA (ssDNA) overhangs, which are rapidly coated by RecA recombinases Dmc1 and Rad51. This nucleoprotein filament then seeks out homology and invades a homologous template, forming a metastable single-end invasion intermediate (D-loop) (3). The invading strand primes DNA synthesis and extends the D-loop. If the extended D-loop is captured by ssDNA on the other side of DSBs in a process known as second-end capture, a double Holliday junction (dHJ) forms (4). While dHJs can be resolved biochemically as either crossovers or non–crossovers (5), during meiosis, the majority of dHJs are specifically resolved as crossovers through the activity of MutLγ (MLH1-MLH3) (6–8) or other structure-selective endonucleases. Although a multitude of DSBs are generated during meiotic prophase, strikingly few are ultimately selected to become crossovers. Early recombination intermediates pare down in pachytene until each homolog pair receives at least one crossover, while the majority of DSBs are repaired as non–crossovers via synthesis-dependent strand annealing (9). However, how meiotic DSBs are chosen to become crossovers remains poorly understood.Throughout eukaryotes, crossover recombination is primarily controlled by a group of proteins collectively termed “ZMM” (10). Notably, homologs of the yeast RING (Really interesting new gene) domain protein Zip3 [ZHP-1, ZHP-2, ZHP-3, and ZHP-4 in Caenorhabditis elegans (11–14), Drosophila Vilya and Narya/Nenya (15, 16), Hei10 in Arabidopsis (17), and Hei10 and RNF212 in mammals (18, 19)] initially localize as abundant foci or long stretches along the synaptonemal complex (SC) but eventually concentrate at crossover sites in late pachytene (20). These SUMO or ubiquitin ligases appear to promote crossover designation by stabilizing the ZMM proteins at crossover sites while removing them from other recombination intermediates (13, 14, 18, 19, 21, 22). Although many meiotic proteins are shown to be SUMO modified (23), key targets of the Zip3 family proteins remain largely unknown.The meiosis-specific MutS homologs MSH4 and MSH5 form a heterodimeric MutSγ complex and play essential roles in crossover formation in diverse eukaryotes (24–31). MutSγ localizes to recombination intermediates as numerous foci but ultimately accumulates at sites that are destined to become crossovers (32, 33). Biochemical analyses using recombinant MSH4 and MSH5 have shown that MutSγ recognizes single-end invasion intermediates and HJs in vitro (34, 35). HJs activate the ATP hydrolysis of MutSγ and promote the exchange of bound ADP for ATP, inducing the formation of a sliding clamp that dissociates from HJs (35, 36). By iterative loading and embracing DNA duplexes within a dHJ, MutSγ is thought to stabilize crossover–specific recombination intermediates (33, 35). In addition, MutSγ recruits and activates the resolvase activity of MutLγ, enabling biased processing of dHJs into crossovers during meiosis (37, 38).A genetic screen in C. elegans identified a cyclin-like protein COSA-1 as a component essential for processing meiotic DSBs into crossovers (39). The mammalian ortholog CNTD1 was subsequently identified (40), and both COSA-1 and CNTD1 have been shown to localize to crossover sites (39, 41, 42). In the absence of COSA-1/CNTD1, MutSγ components persist as numerous foci in pachytene, and crossover formation is eliminated or severely compromised (33, 40), demonstrating a crucial role of COSA-1/CNTD1 in converting early recombination intermediates into crossovers. Because both COSA-1 and CNTD1 are members of the cyclin family, it is plausible that they form a complex with a cyclin-dependent kinase (CDK) and regulate the recombination process through phosphorylation.Several lines of evidence have suggested that CDK2 might be a relevant kinase partner for CNTD1. CDK2 interacts with CNTD1 in yeast-two hybrid assays (41, 42) and localizes to interstitial chromosome sites (43, 44) in a CNTD1-dependent manner (40). Reduced CDK2 activity leads to a failure in crossover formation, while a hyperactive form of CDK2 causes an increased number of MLH1 foci (45). However, due to its requirement at telomeres in tethering chromosomes to the nuclear envelope, deletion of CDK2 leads to severe defects in SC assembly between paired homologs (synapsis) and pachytene arrest (46–49). Further, while a full-length CNTD1-specific protein of the excepted size was detected (using CNTD1 antibodies) in one study (42), a short CNTD1 isoform that cannot interact with CDK2 was the predominant isoform detected in another study (using hemagglutinin antibodies in Cntd1^FH/FH mice with an epitope tag sequence inserted into the endogenous Cntd1 locus) (41), raising questions regarding the extent to which CDK2 and CNTD1 might act as functional partners. Thus, it has been difficult to determine the role of CDK2 in crossover recombination. Moreover, key meiotic targets of CDK2 have not yet been identified.We reasoned that CDK-2, the C. elegans homolog of CDK2, might also localize and function at crossover sites. However, global knockdown of C. elegans CDK-2 by RNA interference leads to cell cycle arrest of mitotically proliferating germ cells (50), thereby precluding the analysis of its requirement during meiotic prophase. To overcome this limitation and establish the meiotic function of CDK-2, we use the auxin-inducible degradation system to deplete CDK-2 from the adult germline, demonstrating that CDK-2 partners with COSA-1 to promote crossover formation during C. elegans meiosis. Moreover, we identify MSH-5 as a key substrate for CDK-2 and provide evidence that CDK-2 and COSA-1 partner to promote crossover designation through phosphorylation and activation of the MutSγ complex.     

A Successful Case of Hepatocellular Carcinoma Treated with Atezolizumab Plus Bevacizumab with Multisystem Immune-related Adverse Events

A 63-year-old man with hepatitis C was treated with atezolizumab plus bevacizumab for unresectable diffuse hepatocellular carcinoma (HCC). After four cycles of atezolizumab plus bevacizumab, the diffuse HCC markedly shrank; however, he complained of general fatigue, loss of appetite, and slight loss of muscle strength in the lower legs. He was diagnosed with isolated adrenocorticotropic hormone deficiency (IAD), hypothyroidism, and myopathy, suggesting multisystem immune-related adverse events (irAEs). After administration of hydrocortisone, the clinical symptoms rapidly disappeared. Patients with multisystem irAEs can have favorable outcomes; thus, to continue immune-checkpoint inhibitors therapy, a correct diagnosis and management of multisystem irAEs are important.     

Usefulness of Dapagliflozin for Nephrotic Syndrome Secondary to Diabetic Kidney Disease

An 81-year-old woman with a medical history of type 2 diabetes mellitus and diabetic nephropathy was admitted with a diagnosis of multiple cerebellar infarctions. Proteinuria and leg edema were observed on the day after admission and diagnosed as nephrotic syndrome. Furosemide and spironolactone were started but showed no diuretic effect, and the renal function deteriorated. These agents were then replaced with dapagliflozin, which resulted in a positive diuretic effect and subsequent improvement of hypoalbuminemia and renal dysfunction. This case report demonstrates the utility of dapagliflozin for nephrotic syndrome to achieve a positive diuretic effect and improve hypoalbuminemia without deteriorating the renal function.     

Abutment Coating With Diamond‐Like Carbon Films to Reduce Implant–Abutment Bacterial Leakage

Background: The influence of diamond‐like carbon (DLC) films on bacterial leakage through the interface between abutments and dental implants of external hexagon (EH) and internal hexagon (IH) designs was evaluated. Methods: Film deposition was performed by plasma‐enhanced chemical vapor deposition. Sets of implants and abutments (n = 30 per group, sets of 180 implants) were divided according to connection design and treatment of the abutment base: 1) no treatment (control); 2) DLC film deposition; and 3) Ag‐DLC film deposition. Under sterile conditions, 1 μL Enterococcus faecalis was inoculated inside the implants, and abutments were tightened. The sets were tested for immediate external contamination, suspended in test tubes containing sterile culture broth, and followed for 5 days. Turbidity of the broth indicated bacterial leakage. At the end of the period, the abutments were removed and the internal content of the implants was collected with paper points and plated in Petri dishes. After 24‐hour incubation, they were assessed for bacterial viability and colony‐forming unit counting. Bacterial leakage was analyzed by χ² and Fisher exact tests (α = 5%). Results: The percentage of bacterial leakage was 16.09% for EH implants and 80.71% for IH implants (P <0.0001). The bacterial load was higher inside IH implants (P = 0.000). The type of implant significantly influenced the results (P = 0.000), whereas the films did not (P = 0.487). Conclusion: IH implants show a higher frequency of bacterial leakage; and DLC and Ag‐DLC films do not significantly reduce the frequency of bacterial leakage and bacteria load inside the implants.     

Contact immunotherapy enhances the therapeutic effects of nivolumab in treating in‐transit melanoma: Two cases reports

Because nivolumab significantly prolongs survival in patients with metastatic melanoma, enhancing its antitumor immune response is of great interest to dermato‐oncologists. In this report, we describe two cases of metastatic melanoma successfully treated with nivolumab in combination with contact immunotherapy, using contact sensitizing agents, such as squaric acid dibutylester and diphencyprone. In addition, immunohistochemical staining supported one of the possible mechanisms of this combination therapy. Our present cases suggested a possible therapy for metastatic melanoma using nivolumab in combination with contact immunotherapy.     

Skin gangrene as an extraintestinal manifestation of inflammatory bowel disease

Inflammatory bowel diseases can commonly present many cutaneous lesions which can contribute to the diagnosis of the disease or its activity. The most frequent cutaneous or mucocutaneous manifestations suggesting ulcerative rectocolitis activity are erythema nodosum (3-10%), pyoderma gangrenosum (5-12%) and aphthous stomatitis (4%). Other reactive skin manifestations related to immunological mechanisms associated with the inflammatory bowel disease are: Sweet''s syndrome, arthritis-dermatitis syndrome associated with inflammatory bowel disease and leukocytoclastic vasculitis. We describe the case of a young man with diagnosis of ulcerative rectocolitis, which presented an extensive cutaneous gangrene secondary to microvascular thrombosis. The case represents a dermatologic rarity and should be recognized as a cutaneous manifestation related to the hypercoagulability state observed in the disease''s activity.     

Anti-inflammatory effects of embelin in A549 cells and human asthmatic airway epithelial tissues

Objective: Allergic asthma is the most common type in asthma, which is defined as a chronic inflammatory disease of the lung. In this study, we investigated whether embelin (Emb), the major component of Ardisia japonica BL. (AJB), exhibits anti-inflammatory effects on allergic asthma via inhibition of NF-κB activity using A549 cells and asthmatic airway epithelial tissues.

Methods: Inflammation was induced in A549 cells, a human airway epithelial cell line, by IL-1β (10?ng/ml) treatment for 4?h. The effects of Emb on NF-κB activity and COX-2 protein expression in inflamed airway epithelial cells and human asthmatic airway epithelial tissues were analyzed via western blot. The secretion levels of NF-κB-mediated cytokines/chemokines, including IL-4, 6, 9, 13, TNF-α and eotaxin, were measured by a multiplex assay.

Results: Emb significantly blocked NF-κB activity in IL-1β-treated A549 cells and human asthmatic airway epithelial tissues. COX-2 expression was also reduced in both IL-1β-treated A549 cells and asthmatic tissues Emb application. Emb significantly reduced the secretion of IL-4, IL-6 and eotaxin in human asthmatic airway epithelial tissues by inhibiting activity of NF-κB.

Conclusions: The results of this study suggest that Emb may be used as an anti-inflammatory agent via inhibition of NF-κB and related cytokines.     

Case of erythema nodosum associated with granulomatous mastitis probably due to Corynebacterium infection

Granulomatous mastitis (GM) is a rare chronic inflammatory breast disease that can be mistaken for a variety of inflammatory and neoplastic disorders of the breast. Erythema nodosum (EN) is a common panniculitis, and is often associated with a variety of diseases, yet coexistence of EN and GM is very rare. In this paper, we present a young Japanese woman with EN associated with GM probably due to Corynebacterium infection. Both lesions of EN and GM were successfully treated with oral minocycline and surgical drainage. To the best of our knowledge, this is the first case of EN associated with GM due to Corynebacterium infection in the English‐language published work.