Auxiliary roles of nardilysin in the early diagnosis of acute coronary syndrome: a prospective cohort study, the Nardi-ACS study.

Acute coronary syndrome (ACS) includes myocardial infarction (MI) and unstable angina (UA). MI is defined by elevated necrosis markers, preferably high-sensitivity cardiac troponins (hs-cTn). However, it takes hours for cTn to become elevated after coronary occlusion; therefore, difficulties are associated with diagnosing early post-onset MI or UA. The aim of this prospective cohort study was to examine the diagnostic ability of serum nardilysin (NRDC) for the early detection of ACS. This study consisted of two sequential cohorts, the Phase I cohort, 435 patients presenting to the emergency room (ER) with chest pain, and the Phase II cohort, 486 patients with chest pain who underwent coronary angiography. The final diagnosis was ACS in 155 out of 435 patients (35.6%) in the phase I and 418 out of 486 (86.0%) in the phase II cohort. Among 680 patients who presented within 24 h of onset, 466 patients (68.5%) were diagnosed with ACS. Serum NRDC levels were significantly higher in patients with ACS than in those without ACS. The sensitivity of NRDC in patients who presented within 6 h after the onset was higher than that of hsTnI, and the AUC of NRDC within 1 h of the onset was higher than that of hsTnI (0.718 versus 0.633). Among hsTnI-negative patients (300 of 680 patients: 44.1%), 136 of whom (45.3%) were diagnosed with ACS, the sensitivity and the NPV of NRDC were 73.5 and 65.7%, respectively. When measured in combination with hsTnI, NRDC plays auxiliary roles in the early diagnosis of ACS.

Validation of the severe COVID-19 prognostic value of serum IL-6, IFN-λ3, CCL17, and calprotectin considering the timing of clinical need for prediction.

Although biomarkers to predict coronavirus disease 2019 (COVID-19) severity have been studied since the early pandemic, no clear guidelines on using them in clinical practice are available. Here, we examined the ability of four biomarkers to predict disease severity using conserved sera from COVID-19 patients who received inpatient care between January 1, 2020 and September 21, 2021 at the National Center for Global Health and Medicine, collected at the appropriate time for prediction. We predicted illness severity in two situations: 1) prediction of future oxygen administration for patients without oxygen support within 8 days of onset (Study 1) and 2) prediction of future mechanical ventilation support (excluding non-invasive positive pressure ventilation) or death of patients within 4 days of the start of oxygen administration (Study 2). Interleukin-6, IFN-λ3, thymus and activation-regulated chemokine, and calprotectin were measured retrospectively. Other laboratory and clinical information were collected from medical records. AUCs were calculated from ROC curves and compared for the predictive ability of the four biomarkers. Study 1 included 18 patients, five of whom had developed oxygen needs. Study 2 included 45 patients, 13 of whom required ventilator management or died. In Study 1, IFN-λ3 showed a good predictive ability with an AUC of 0.92 (95% CI 0.76-1.00). In Study 2, the AUC of each biomarker was 0.70-0.74. The number of biomarkers above the cutoff showed the possibility of good prediction with an AUC of 0.86 (95% CI 0.75-0.97). When two or more biomarkers were positive, sensitivity and specificity were 0.92 and 0.63, respectively. In terms of biomarker testing at times when prognostication may be clinically useful, IFN-λ3 was predictive of oxygenation demand and a combination of the four biomarkers was predictive of mechanical ventilator requirement.

Less Invasive Management of Tissue Deficits for Deep Sternal Wound Infections.

BACKGROUND: The frequency of sternomyelitis after cardiovascular surgery has been reported to be 0.4% -5%. METHODS: The treatment method used for 47 patients (29 male and 18 female) who developed sternomyelitis after sternotomy with tissue defects in the chest was examined retrospectively. RESULTS: Of the original conditions, the most frequent was coronary artery disease undergoing bypass grafting (22 cases, 46.8%), followed by acute aortic dissection (10 cases, 21.3%). The number of times debridement was performed was: once, 35 cases; twice, 11 cases; 7 times, 1 case; and unknown, 2 cases. The most frequent time of occurrence of sternomyelitis was within 2 weeks after surgery (12 patients, 25.5%). A residual internal thoracic artery remained on both sides in 28 cases (59.6%), and only on the right side in 17 cases (36.2%); there was no remaining one in 2 cases (4.2%). The reconstruction method was a pectoralis major musculocutaneous flap in 31 cases (66.0%), internal mammary artery perforator flap in 7 cases (14.9%), rectus abdominis musculocutaneous flap in 4 cases (8.5%), omentum transplant in 3 cases (6.4%), superior epigastric artery perforator flap in 2 cases (4.3%), external abdominal oblique muscle flap in 1 case (2.1%), and latissimus dorsi musculocutaneous flap in 1 case (2.1%). The internal mammary artery perforator flap and the superior epigastric artery perforator flap have been effective treatment. CONCLUSIONS: In 47 patients, our method of treatment for tissue defects of the chest wall after sternal osteomyelitis was examined, and an algorithm using less invasive management was proposed.

Serum Nardilysin, a Surrogate Marker for Epithelial-Mesenchymal Transition, Predicts Prognosis of Intrahepatic Cholangiocarcinoma after Surgical Resection.

PURPOSE: Few studies have investigated prognostic biomarkers in patients with intrahepatic cholangiocarcinoma (ICC). Nardilysin (NRDC), a metalloendopeptidase of the M16 family, has been suggested to play important roles in inflammation and several cancer types. We herein examined the clinical significance and biological function of NRDC in ICC.Experimental Design: We measured serum NRDC levels in 98 patients with ICC who underwent surgical resection in two independent cohorts to assess its prognostic impact. We also analyzed NRDC mRNA levels in cancerous tissue specimens from 43 patients with ICC. We investigated the roles of NRDC in cell proliferation, migration, gemcitabine sensitivity, and gene expression in ICC cell lines using gene silencing. RESULTS: High serum NRDC levels were associated with shorter overall survival and disease-free survival in the primary (n = 79) and validation (n = 19) cohorts. A correlation was observed between serum protein levels and cancerous tissue mRNA levels of NRDC (Spearman ρ = 0.413; P = 0.006). The gene knockdown of NRDC in ICC cell lines attenuated cell proliferation, migration, and tumor growth in xenografts, and increased sensitivity to gemcitabine. The gene knockdown of NRDC was also accompanied by significant changes in the expression of several epithelial-mesenchymal transition (EMT)-related genes. Strong correlations were observed between the mRNA levels of NRDC and EMT-inducing transcription factors, ZEB1 and SNAI1, in surgical specimens from patients with ICC. CONCLUSIONS: Serum NRDC, a possible surrogate marker reflecting the EMT state in primary tumors, predicts the outcome of ICC after surgical resection.

In Situ Splitting of a Rib Bone Graft for Reconstruction of Orbital Floor and Medial Wall.

In situ splitting of rib bone graft was conducted in 22 patients for the repair of orbital fracture with no other complicating fractures. A bone graft was harvested from the sixth or seventh rib in the right side. The repair of the orbital floor and medial wall was successful in all the cases. Ten patients had bone grafting to the orbital floor, eight had it done onto medial wall, and 4 onto both floor and wall after reduction. The mean length of in situ rib bone graft was 40.9 mm (range, 20-70 mm), the mean width of these was 14.9 mm (range, 8-20 mm). The bone grafting was done by one leaf for 15 cases and two leafs for 7 cases in size of defects. The technique of in situ splitting of a rib bone graft for the repair of the orbital floor and medial wall is a simple and safe procedure, easily taking out the in situ splitting of a rib, and less pain in donor site. It has proved to be an optimal choice in craniofacial reconstruction, especially the defects of orbital floor and medial wall.

Nardilysin promotes hepatocellular carcinoma through activation of signal transducer and activator of transcription 3.

Nardilysin (NRDC) is a metalloendopeptidase of the M16 family. We previously showed that NRDC activates inflammatory cytokine signaling, including interleukin-6-signal transducer and activator of transcription 3 (STAT3) signaling. NRDC has been implicated in the promotion of breast, gastric and esophageal cancer, as well as the development of liver fibrosis. In this study, we investigated the role of NRDC in the promotion of hepatocellular carcinoma (HCC), both clinically and experimentally. We found that NRDC expression was upregulated threefold in HCC tissue compared to the adjacent non-tumor liver tissue, which was confirmed by immunohistochemistry and western blotting. We also found that high serum NRDC was associated with large tumor size (>3 cm, P = 0.016) and poor prognosis after hepatectomy (median survival time 32.0 vs 73.9 months, P = 0.003) in patients with hepatitis C (n = 120). Diethylnitrosamine-induced hepatocarcinogenesis was suppressed in heterozygous NRDC-deficient mice compared to their wild-type littermates. Gene silencing of NRDC with miRNA diminished the growth of Huh-7 and Hep3B spheroids in vitro. Notably, phosphorylation of STAT3 was decreased in NRDC-depleted Huh-7 spheroids compared to control spheroids. The effect of a STAT3 inhibitor (S3I-201) on the growth of Huh-7 spheroids was reduced in NRDC-depleted cells relative to controls. Our results show that NRDC is a promising prognostic marker for HCC in patients with hepatitis C, and that NRDC promotes tumor growth through activation of STAT3.

Comparison of malignant skin tumor thickness and relative depth of invasion estimates from preoperative MR-microscopy and pathological evaluation.

BACKGROUND: Tumor thickness and relative depth of invasion are prognostic parameters considered when developing treatment plans for malignant skin tumors. Although conventional magnetic resonance (MR) imaging techniques cannot identify small tumors, use of microscopy coils considerably improves spatial resolution. Some studies have shown that this technique is efficacious in preoperative assessment of relative depth of invasion; however, its ability to provide accurate measurements of tumor thickness remains unconfirmed. OBJECTIVE: The purpose of this pilot study was to evaluate the usefulness of preoperative MR-microscopy in determining tumor thickness and relative depth of invasion of malignant skin tumors. METHODS AND MATERIALS: Magnetic resonance images of malignant skin tumors in seven female patients (six with basal cell carcinoma and one with malignant melanoma) were obtained using a 1.5 T system and a 47-mm or 23-mm microscopy coil. Tumors were then excised, fixed, dehydrated, embedded, and stained with hematoxylin and eosin. We then compared MR-microscopy and pathology values for thickness and relative depth of invasion of each tumor. RESULTS: Both techniques produced similar measurements of tumor thickness and relative depth of invasion. CONCLUSIONS: MR-microscopy is very useful for accurate preoperative estimation of not only relative depth of invasion, but also thickness of malignant skin tumors.

Growth and poliovirus production of Vero cells on a novel microcarrier with artificial cell adhesive protein under serum-free conditions.

A microcarrier is used for the three-dimensional (3D) culture of adhesion-dependent mammalian cells. We developed a novel microcarrier by binding ProNectin F, an artificial cell adhesive protein synthesized by genetically engineered Escherichia coli to a polyacrylic superabsorbent polymer. The microcarrier is characterized by containing no animal-derived components. The serum-free culture of Vero cells for vaccine production using the microcarrier increased the number of Vero cells by approximately 30% compared with the existing dextran beads coated with porcine Type I collagen, which resulted in approximately a 30% to 40% increase in the infectivity titer of the Sabin 2 strain of poliovirus. These results suggested that the developed microcarrier should be unprecedented in permitting high-yield vaccine production by means of a serum-free culture.

Elastic fiber assembly is disrupted by excessive accumulation of chondroitin sulfate in the human dermal fibrotic disease, keloid.

Keloid is a fibrotic disease characterized by abnormal accumulation of extracellular matrix in the dermis. The keloid matrix contains excess collagen and glycosaminoglycans (GAGs), but lacks elastic fiber. However, the roles of these matrix components in the pathogenesis of keloid are largely unknown. Here, we show that elastin and DANCE (also known as fibulin-5), a protein required for elastic fiber formation, are not deposited in the extracellular matrix of keloids, due to excess accumulation of chondoitin sulfate (CS), although the expression of elastin and DANCE is not affected. Amount of CS accumulated in the keloid legion was 6.9-fold higher than in normal skin. Fibrillin-1, a scaffold protein for elastic fiber assembly, was abnormally distributed in the keloid matrix. Addition of purified CS to keloid fibroblast culture resulted in abnormal deposition of fibrillin-1, concomitant with significantly decreased accumulation of elastin and DANCE in the extracellular matrix. We propose that CS plays a crucial role in the development of keloid lesions through inhibition of elastic fiber assembly.

Silklike materials constructed from sequences of Bombyx mori silk fibroin, fibronectin, and elastin.

Two silklike proteins, [TGRGDSPAGG(GAGAGS)3AS]5 (FS5) and [TGRGDSPA-(GVPGV)2GG(GAGAGS)3AS]8 (FES8) were designed to demonstrate the superior performance as biomaterials of silklike proteins. The former protein consists of the crystalline domain sequence, (GAGAGS)n from Bombyx mori silk fibroin and cell-adhesive sequence TGRGDSPA coming from fibronectin-containing RGD triplet. The additional sequence (GVPGV)n from elastin was included in the latter protein. The considerably higher cell-adhesion activities of these proteins for NHDF and VERO cells were observed by comparing with those of silklike materials without RGD sequences and also the crystalline fraction of B. mori silk fibroin. This tendency was independent of the treatments, 4.5M LiClO4 or formic acid (FA), on silklike proteins. Their activities are also higher than those of commercial Fibronectin F for NHDF cell. Their structural characterization was studied using 13C solid-state NMR. Although the overlapped peaks in usual 13C CP/MAS NMR spectra make the detailed structural analysis difficult, the methyl resonance regions observed using dipolar dephasing NMR were very useful for the analysis. The presence of both random coil and beta-sheet structures was observed in these proteins clearly. The content of beta-sheet structure in both proteins increases after FA treatment when compared with the lyophilized samples. The production of electrospun nanofibers from their hexafluoroacetone solution was also tried. The silklike protein FES8 could prepare nonwoven silk fibers although FS5 could not.

Design of silk-like biomaterials inspired by mussel-adhesive protein.

To develop biomaterials for tissue engineering, a silk-like protein inspired by mussel-adhesive proteins (MAPs) was designed and prepared. The primary structure of this silk-like protein is designed as TS[AKPSYPPTYKAS (GAGAGS)(3)](10) by combining the sequences (GAGAGS)(3), the crystalline region of Bombyx mori silk fibroin, and AKPSYPPTYK, the adhesive sequence of MAP from Mytilus edulis. This protein was synthesized by the genetic engineering method. Solid-state (13)C NMR spectra showed that this silk-like protein adopts flexible conformation due to introduction of the sequence AKPSYPPTYK. Cell assay indicated that this silk-like protein has significantly higher cell adhesion activities in response to normal human dermal fibroblasts (NHDFs) than Pronectin F, which is available as commercialized cell-adhesive silk-like protein. Thus, combination of remarkably high cell-adhesive activity from MAP with superiority of silk fibroin provides potentiality for application to the field of biomaterials.

Nonmicrosurgical replantation using a subcutaneous pocket for salvage of the amputated fingertip.

PURPOSE: The pocket principle suggested by Brent in 1979 is an alternative method for use when microsurgical replantation is not feasible. The application and the amputation level for which the method is available, however, have not been well examined. METHODS: Between 1999 and 2003 we treated 6 patients (7 fingers) by nonmicrosurgical replantation using a subcutaneous pocket (the Brent technique). All patients had sustained complete fingertip amputations across or proximal to the lunula in digits other than the thumb. In every case the amputation was a crush or avulsion-type injury and microsurgical replantation was not feasible; however, cosmetic symmetry was desired strongly by the patient. RESULTS: Of the 7 fingers only one survived completely but became atrophic after 4 months. One finger developed necrosis involving less than half of the replant but a hooked nail deformity developed. Two fingers developed partial necrosis involving more than half of the replant but both fingers were missing the fingernail and the cosmetic results were not acceptable. Three fingers developed total necrosis. In addition a slight flexion contracture not improved with therapy in the digits was noted in 4 patients. CONCLUSIONS: The Brent technique should be performed scrupulously for fingertip amputation across or proximal to the lunula because of the poor survival rate and the possibility of contracture in the digits or other proximal joints.

The PNB classification for treatment of fingertip injuries: the boundary between conservative treatment and surgical treatment.

The PNB classification, which was advocated by Evans and Bernadis, separates the injuries into their effects on 3 components of the fingertip: pulp, nail, and bone. Because each component is subdivided into 7 or 8 items, this can describe fingertip injuries more precisely. Between 1997 and 2003, we treated 381 fingertip injuries (279 males, 102 females; average age, 41.2 years) in our facilities. A 3-digit number was provided for each of the 381 cases in accordance with the PNB classification. We extracted patients in whom amputated tissues did not exist, and predicted the boundary between conservative treatment and surgical treatment by individually comparing the curative results of the same type of injuries. In conclusion, PNB 355-366 and PNB 455-466 were most suitable for surgical treatment, and the boundaries between surgical treatment and conservative treatment were PNB 386 and 666 and 700. The results, which are the criteria for surgical treatment, are summarized as follows; 1) More than two thirds of the distal phalanx remains. 2)The nail bed defect ranges from one third to half. If the defect is more or less than the criteria, the surgical treatment is less significant. Recognition of the boundary and prevention from unnecessary surgical treatment leads to minimum invasive surgery for fingertip injuries.

A method for correcting an inverted nipple with an artificial dermis.

Various methods have been reported to correct an inverted nipple. Although a satisfactory outcome has been reported with most techniques, each method carries a drawback inherent in the technique itself, including complicated operative technique, sensory disturbance of the nipple, marked scarring of the nipple areola and other donor regions, destruction of breast function, and incomplete correction. This report describes a simple method for correcting an inverted nipple. It incorporates a new concept of using artificial dermis for tissue augmentation and is performed without sacrificing any donor site and complex design. It was applied to four nipples in two nulliparous cases. For all four corrected inverted nipples, good results were obtained, and there have been no complications. There were no deformities of the nipples or the areolas after this procedure, and the surgical scars were inconspicuous.

Production and characterization of a silk-like hybrid protein, based on the polyalanine region of Samia cynthia ricini silk fibroin and a cell adhesive region derived from fibronectin.

There are a variety of silkworms and silk fibroins produced by them. Silks have many inherent suitable properties for biomaterials. In this paper, a novel silk-like hybrid protein, [DGG(A)(12)GGAASTGRGDSPAAS](5), which consists of polyalanine region of silk fibroin from a wild silkworm, Samia cynthia ricini, and cell adhesive region including Arg-Gly-Asp (RGD) sequence, derived from fibronectin, was designed and produced. The genes encoding the hybrid protein were constructed and expressed in Escherichia coli. The main conformation of the polyalanine region, that is, either alpha-helix or beta-sheet, could be easily controlled by treatment with different acidic solvents, trifluoroacetic acid or formic acid, respectively. This structural change was monitored with 13C CP/MAS NMR. Higher cell adhesive and growth activities of the hybrid protein compared with those of collagen were obtained.

Successful replantation of an amputated leg with severe crush and avulsion injury in an elderly patient: case report.

An amputated leg with severe crush and avulsion injury was replanted in a 57-year-old man. Since the skin and soft tissue around the ankle became necrotic 11 days postoperatively, a flow-through forearm free flap was transferred into the defect, preserving the blood circulation to the replanted leg. Seven weeks after the replantation, sural nerve cable grafts were interposed between both amputated edges of the tibial nerve. As the function of the ankle joint did not recover, arthrodesis of the ankle joint was performed 7 months after the replantaion. As a result, the patient could walk 1 year postoperatively.