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Abstract To evaluate the antibody response following the initial four doses of mRNA vaccines (BNT162b2 or mRNA-1273) in SARS-CoV-2-naïve healthy adults and investigate factors influencing antibody titer increases, this prospective cohort study was conducted in Japan from March 2021. The study included participants who received either the 1st and 2nd doses (n = 467), 3rd dose (n = 157), or 4th dose (n = 89). Blood samples were collected before and up to 6 months after each dose, and anti-receptor-binding domain antibody levels were measured. Multivariate analysis (usin multiple linear regression or linear mixed models) revealed several factors significantly associated with higher post-vaccination antibody levels, including mRNA-1273 vaccine (after the 1st and 2nd dose), male gender (after the 3rd and 4th doses), younger age (after the 1st and 2nd dose), non-smoking status (after the 2nd dose), non-use of immunosuppressive agents (after the 1st dose), higher pre-vaccination antibody titers (after the 2nd, 3rd, and 4th doses), and higher post-vaccination fever (after the 2nd and 4th doses). Furthermore, longer intervals since the last dose were significantly associated with higher antibody levels after the 3rd and 4th doses. These findings provide valuable insights for optimizing vaccination strategies.

Abstract Background Adults infected with Plasmodium spp. in endemic areas need to be re-evaluated in light of global malaria elimination goals. They potentially undermine malaria interventions but remain an overlooked aspect of public health strategies. Methods This study aimed to estimate the prevalence of Plasmodium spp. infections, to identify underlying parasite species, and to assess predicting factors among adults residing in an endemic area from the Democratic Republic of Congo (DRC). A community-based cross-sectional survey in subjects aged 18 years and above was therefore carried out. Study participants were interviewed using a standard questionnaire and tested for Plasmodium spp. using a rapid diagnostic test and a nested polymerase chain reaction assay. Logistic regression models were fitted to assess the effect of potential predictive factors for infections with different Plasmodium spp. Results Overall, 420 adults with an estimated prevalence of Plasmodium spp. infections of 60.2% [95% CI 55.5; 64.8] were included. Non-falciparum species infected 26.2% [95% CI 22.2; 30.5] of the study population. Among infected participants, three parasite species were identified, including Plasmodium falciparum (88.5%), Plasmodium malariae (39.9%), and Plasmodium ovale (7.5%) but no Plasmodium vivax. Mixed species accounted for 42.3% of infections while single-species infections predominated with P. falciparum (56.5%) among infected participants. All infected participants were asymptomatic at the time of the survey. Adults belonging to the “most economically disadvantaged” households had increased risks of infections with any Plasmodium spp. (adjusted odds ratio, aOR = 2.87 [95% CI 1.66, 20.07]; p

Abstract Background Chagas disease can lead to life-threatening cardiac manifestations. Regional factors, including genetic characteristics of circulating Trypanosoma cruzi (T. cruzi), have attracted attention as likely determinants of Chagas disease phenotypic expression and Chagas cardiomyopathy (CCM) progression. Our objective was to elucidate the differential transcriptomic signatures of cardiomyocytes resulting from infection with genetically discrete T. cruzi strains and explore their relationships with CCM pathogenesis and progression. Methods HL-1 rodent cardiomyocytes were infected with T. cruzi trypomastigotes of the Colombian, Y, or Tulahuen strain. RNA was serially isolated post-infection for microarray analysis. Enrichment analyses of differentially expressed genes (fold-change ≥ 2 or ≤ 0.5) highlighted over-represented biological pathways. Intracellular levels of reactive oxygen species (ROS) were compared between T. cruzi-infected and non-infected HL-1 cardiomyocytes. Results We found that oxidative stress-related gene ontology terms (GO terms), ‘Hypertrophy model’, ‘Apoptosis’, and ‘MAPK signaling’ pathways (all with P

Abstract Context The Democratic Republic of Congo (DRC), one of the most malaria-affected countries worldwide, is a potential hub for global drug-resistant malaria. This study aimed at summarizing and mapping surveys of malaria parasites carrying molecular markers of drug-resistance across the country. Methods A systematic mapping review was carried out before July 2023 by searching for relevant articles through seven databases (PubMed, Embase, Scopus, African Journal Online, African Index Medicus, Bioline and Web of Science). Results We identified 1541 primary studies of which 29 fulfilled inclusion criteria and provided information related to 6385 Plasmodium falciparum clinical isolates (collected from 2000 to 2020). We noted the PfCRT K76T mutation encoding for chloroquine-resistance in median 32.1% [interquartile interval, IQR: 45.2] of analyzed malaria parasites. The proportion of parasites carrying this mutation decreased overtime, but wide geographic variations persisted. A single isolate had encoded the PfK13 R561H substitution that is invoked in artemisinin-resistance emergence in the Great Lakes region of Africa. Parasites carrying various mutations linked to resistance to the sulfadoxine–pyrimethamine combination were widespread and reflected a moderate resistance profile (PfDHPS A437G: 99.5% [IQR: 3.9]; PfDHPS K540E: 38.9% [IQR: 47.7]) with median 13.1% [IQR: 10.3] of them being quintuple IRN–GE mutants (i.e., parasites carrying the PfDHFR N51I–C59R–S108N and PfDHPS A437G–K540E mutations). These quintuple mutants tended to prevail in eastern regions of the country. Among circulating parasites, we did not record any parasites harboring mutations related to mefloquine-resistance, but we could suspect those with decreased susceptibility to quinine, amodiaquine, and lumefantrine based on corresponding molecular surrogates. Conclusions Drug resistance poses a serious threat to existing malaria therapies and chemoprevention options in the DRC. This review provides a baseline for monitoring public health efforts as well as evidence for decision-making in support of national malaria policies and for implementing regionally tailored control measures across the country.

More people with a history of prior infection are receiving SARS-CoV-2 vaccines. Understanding the level of protection granted by ‘hybrid immunity’, the combined response of infection- and vaccine-induced immunity, may impact vaccination strategies through tailored dosing. A total of 36 infected (‘prior infection’) and 33 SARS-CoV-2 ‘naïve’ individuals participated. Participants provided sera six months after completing a round of BNT162b2 vaccination, to be processed for anti-spike antibody measurements and the receptor binding domain-ACE2 binding inhibition assays. The relationships between antibody titer, groups and age were explored. Anti-spike antibody titers at 6 months post-vaccination were significantly higher, reaching 13- to 17-fold, in the ‘prior infection’ group. Semi-log regression models showed that participants with ‘prior infection’ demonstrated higher antibody titer compared with the ‘naïve’ even after adjusting for age. The enhancement in antibody titer attributable to positive infection history increased from 8.9- to 9.4- fold at age 30 to 19- to 32-fold at age 60. Sera from the ‘prior infection’ group showed higher inhibition capacity against all six analyzed strains, including the Omicron variant. Prior COVID-19 led to establishing enhanced humoral immunity at 6 months after vaccination. Antibody fold-difference attributed to positive COVID-19 history increased with age, possibly because older individuals are prone to symptomatic infection accompanied by potentiated immune responses. While still pending any modifications of dosing recommendations (i.e. reduced doses for individuals with prior infection), our observation adds to the series of real-world data demonstrating the enhanced and more durable immune response evoked by booster vaccinations following prior infection.

This study assessed the immunogenicity and safety of the BNT162b2 mRNA vaccine in lung cancer patients receiving anticancer treatment. We enrolled lung cancer patients receiving anticancer treatment and non-cancer patients; all participants were fully vaccinated with the BNT162b2 vaccine. Blood samples were collected before the first and second vaccinations and 4 ± 1 weeks after the second vaccination. Anti-severe respiratory syndrome coronavirus-2 (SARS-CoV-2) spike protein S1 subunit receptor-binding domain antibody titers were measured using the Architect SARS-CoV-2 IgG II Quant and Elecsys Anti-SARS-CoV-2 S assays. Fifty-five lung cancer patients and 38 non-cancer patients were included in the immunogenicity analysis. Lung cancer patients showed significant increase in the geometric mean antibody concentration, which was significantly lower than that in the non-cancer patients after the first (30 vs. 121 AU/mL, p

ABSTRACT Past severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is an important determinant of protection from reinfection and of postvaccine immune responses. Herein, we conducted a follow-up analysis of health care workers previously infected with coronavirus disease 2019 (COVID-19) with the aim of evaluating different immunoassays for their capability in detecting the waning anti-SARS-CoV-2 immune responses and accuracy in documenting past SARS-CoV-2 infections. We evaluated serum antinucleocapsid antibody levels in convalescent individuals following a 1.5-year interval from SARS-CoV-2 infection. Three different commercial immunoassays that qualitatively measure serum antibodies targeting the SARS-CoV-2 nucleocapsid protein, namely, the Abbott Architect SARS-CoV-2 IgG, the Euroimmun anti-SARS-CoV-2 NCP enzyme-linked immunosorbent assay (ELISA) IgG, and the Roche Elecsys anti-SARS-CoV-2, were tested for comparison of detectability. A total of 38 individuals consented to participating in this follow-up analysis. From assay to assay, seropositivity rate at 18 months from infection varied from lowest at 42% to highest at 92%. The Roche Elecsys immunoassay, dependent on the dual-antigen antibody detection method and tuned for the detection of high avidity antibodies, was most capable of accurately documenting past SARS-CoV-2 infections. Different immunoassays showed variable capability of determining previous infection status under waning antibody concentrations. Immunoassays with lower detection limits are to be selected, and adjusted thresholds are to be considered in order to maximize the tests’ performance. IMPORTANCE Past SARS-CoV-2 infection is an important determinant of protection from reinfection and of postvaccine immune responses. Our results show that different immunoassays, by design, harbor variable capability of tracking SARS-CoV-2 infection under waning antibody concentrations. With each recovered patient standing at a unique time point along the decline curve of antibodies, precise estimation of COVID-19 cumulative incidence remains a challenge. Since future surveillance studies will be targeting more than ever heterogenous cohorts, selecting the appropriate immunoassay is crucial in order to assure reliable decisions about an individual’s previous infection status.

ABSTRACT The prompt rollout of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mRNA vaccine is facilitating population immunity, which is becoming more dominant than natural infection-mediated immunity. In the midst of coronavirus disease 2019 (COVID-19) vaccine deployment, understanding the epitope profiles of vaccine-elicited antibodies will be the first step in assessing the functionality of vaccine-induced immunity. In this study, the high-resolution linear epitope profiles of Pfizer-BioNTech COVID-19 mRNA vaccine recipients and COVID-19 patients were delineated by using microarrays mapped with overlapping peptides of the receptor binding domain (RBD) of the SARS-CoV-2 spike protein. The vaccine-induced antibodies targeting the RBD had a broader distribution across the RBD than that induced by the natural infection. Half-maximal neutralization titers were measured in vitro by live virus neutralization assays. As a result, relatively lower neutralizability was observed in vaccine recipient sera, when normalized to a total anti-RBD IgG titer. However, mutation panel assays targeting the SARS-CoV-2 variants of concern have shown that the vaccine-induced epitope variety, rich in breadth, may grant resistance against future viral evolutionary escapes, serving as an advantage of vaccine-induced immunity. IMPORTANCE Establishing vaccine-based population immunity has been the key factor in attaining herd protection. Thanks to expedited worldwide research efforts, the potency of mRNA vaccines against the coronavirus disease 2019 (COVID-19) is now incontestable. The next debate is regarding the coverage of SARS-CoV-2 variants. In the midst of vaccine deployment, it is of importance to describe the similarities and differences between the immune responses of COVID-19 vaccine recipients and naturally infected individuals. In this study, we demonstrated that the antibody profiles of vaccine recipients are richer in variety, targeting a key protein of the invading virus, than those of naturally infected individuals. Vaccine-elicited antibodies included more nonneutralizing antibodies than infection-elicited antibodies, and their breadth in antibody variations suggested possible resilience against future SARS-CoV-2 variants. The antibody profile achieved by vaccinations in naive individuals provides important insight into the first step toward vaccine-based population immunity.

ABSTRACT We describe the results of testing health care workers, from a tertiary care hospital in Japan that had experienced a coronavirus disease 2019 (COVID-19) outbreak during the first peak of the pandemic, for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific antibody seroconversion. Using two chemiluminescent immunoassays and a confirmatory surrogate virus neutralization test, serological testing revealed that a surprising 42% of overlooked COVID-19 diagnoses (27/64 cases) occurred when case detection relied solely on SARS-CoV-2 nucleic acid amplification testing (NAAT). Our results suggest that the NAAT-positive population is only the tip of the iceberg and the portion left undetected might potentially have led to silent transmissions and triggered the spread. A questionnaire-based risk assessment was further indicative of exposures to specific aerosol-generating procedures (i.e., noninvasive ventilation and airway suctioning) having mediated transmission and served as the origins of the outbreak. Our observations are supportive of a multitiered testing approach, including the use of serological diagnostics, in order to accomplish exhaustive case detection along the whole COVID-19 spectrum. IMPORTANCE We describe the results of testing frontline health care workers, from a hospital in Japan that had experienced a COVID-19 outbreak, for SARS-CoV-2-specific antibodies. Antibody testing revealed that a surprising 42% of overlooked COVID-19 diagnoses occurred when case detection relied solely on PCR-based viral detection. COVID-19 clusters have been continuously striking the health care system around the globe. Our findings illustrate that such clusters are lined with hidden infections eluding detection with diagnostic PCR and that the cluster burden in total is more immense than actually recognized. The mainstays of diagnosing infectious diseases, including COVID-19, generally consist of two approaches, one aiming to detect molecular fragments of the invading pathogen and the other to measure immune responses of the host. Considering antibody testing as one trustworthy option to test our way through the pandemic can aid in the exhaustive case detection of COVID-19 patients with variable presentations.

Introduction: In severe extremity trauma involving large tissue defects, early closure (e.g., free-flap surgery) of the defects is an essential step for good functional reconstruction; however, in some cases, early closure may be difficult. Highly concentrated carbon dioxide bathing, used to improve blood flow in ischemic limbs and skin ulcers, can also be applied in wound bed preparation for severe limb trauma. Patients and Methods: The three cases in this study required an average of 13 weeks of highly concentrated carbonated bathing, which led to significantly better wound bed preparation, even in the exposed bone and tendon regions. Results: We successfully achieved good functional limb reconstruction in patients with deep burns and severe open fractures by reducing wound infection and facilitating good wound bed preparation. Conclusions: Highly concentrated carbon dioxide bathing was sufficient to prevent frequent wound infections, even in severe extremity trauma involving large soft-tissue defects such as deep crush burns and Gustilo Anderson classification ≥3b open fractures of the extremities. To our knowledge, such interventions have not been reported in the past and are valuable as new procedures for wound bed preparation in severe extremity trauma from both cost and wound infection control perspectives.

Objective: To examine morphological changes of the lip and nose during resting and lip protrusion in patients with unilateral cleft lip. Methods: A total of 41 patients with unilateral cleft lip who underwent primary lip repair were included in the study population. The distortion of the lip and nose was measured and evaluated using photographs of the preoperative and postoperative resting lips and the postoperative protruding lips. Results: The position of the labial commissure on the affected side was cranially deviated before lip repair, and the deviation remained after surgery. The degree of cranial deviation was increased by lip protrusion. The position of the alar base on the affected side was caudally deviated before lip repair and improved after surgery; however, it was displaced cranially by lip protrusion. Horizontal displacement of the lip and nose was improved after surgery, and there was no change during lip protrusion. Conclusions: The characteristic facial distortion in patients with unilateral cleft lip were confirmed. Patients with cleft lip had distortions in the labial commissure and the nose. One of the important future challenges in cleft lip treatment is to focus on changes caused by muscle movement and provide natural and symmetrical facial expressions.

Large bilateral symmetrical vulvar tumors are rare. A 30-year-old multiparous Asian woman presented with severe discomfort during urination and intercourse owing to a large bilateral symmetrical lipoma in the labia majora. En bloc resection through a pre-existing Caesarean scar achieved good cosmetic and symptomatic results. This alternative and unique approach may be helpful in similar rare cases.

Background The effect of artificial highly concentrated carbon dioxide bathing on heat-press-injured wounds has been clinically observed. Case presentation A 46-year-old male patient with a heat press injury of the right palmar region achieved complete necrotic tissue removal and sufficient granulation of the skin graft by continuous hand bathing in a solution containing highly concentrated carbon dioxide for four weeks. The patient successfully underwent a full-thickness skin graft from the plantar to the palmar region, and his right hand function improved to the extent that it did not interfere with daily life. Conclusions Heat press injuries are often refractory to treatment owing to the synergistic effects of heat and crush injuries. Highly concentrated carbon dioxide bathing has long been known to increase skin and muscle blood flow, and its clinical applications in ischemic limbs, skin ulcers, and osteomyelitis have already been reported. In this case, the results suggest an effect of improved blood flow leading to maturation of the graft bed at the heat-press injury site. Lay Summary Highly concentrated carbon dioxide bathing is not only clinically applicable to ischemic extremities, skin ulcers, and osteomyelitis, due to increased cutaneous and muscular blood flow, but also to the reliable debridement of heat press injury wounds and the granulation of graft beds. We successfully used this novel combination of procedures to establish a graft bed with a good blood flow despite the deep heat press injury occurring in the palmar region, leading to an excellent functional recovery. The cost per procedure is low, at about $1(USD), making it a wound care option that should be considered from a healthcare economic perspective.

Internal fixation using plates remains the gold standard for facial fracture fixation. External wound fixators are rarely used. In the present study, we successfully treated multiple comminuted facial fractures with midfacial depressions by using a Hoffman-type external fixator and achieved good functional and cosmetic results. Internal fixation using plates is sometimes problematic in comminuted facial fractures, such as those associated with high-energy trauma. Depending on the fracture pattern, a Hoffman-type external fixator, which is a single pillar type of external fixator, can be effective.

Abstract Background In recent years, the number of patients with ischemic skin ulcers due to diabetes mellitus and arteriosclerosis obliterans are increasing. Accordingly, endovascular therapy, drugs, and various wound dressings have been developed and applied to diabetic foot ulcers, and negative-pressure wound therapy, which often requires expensive and burdensome procedures for medical personnel, has also become popular. So simple and minimal invasive home treatment by the patient or their caregiver is required. Case presentation The present patient (77 years old, male, Asian) had developed left sole ulcers with draining pus that were resistant to conventional treatment, and he suffered from gait disturbance. We report a case of metatarsal osteomyelitis in a patient with diabetes mellitus and arteriosclerosis obliterans, in whom artificial carbon dioxide foot bathing and povidone–iodine sugar ointment were used continuously to promote bone and joint regeneration, and skin ulcer healing. Conclusions A simple therapeutic intervention with artificial carbon dioxide foot bathing and povidone–iodine sugar ointment can improve not only ischemic skin ulcers, but also the bone and joint regeneration of ischemic limbs. This therapy can lead to a reduction in healthcare costs for a huge number of diabetic patients.

We herein report a 57-year-old man afflicted with multiple malignant tumors arising from a giant sebaceous nevus on his left parieto-temporal scalp and neck. The upper eyelid was resected in all layers, and the auricle was resected as well, leaving part of the external auricular cartilage; the parotid gland area was also resected over the parotid fascia. The lost part of the left eyebrow was reconstructed using an anterior forehead skin flap, and the residual defect was covered with skin graft and expanded scalp flap. This is probably the first case report of giant sebaceous nevus associated with three malignancies: sebaceous carcinoma, apocrine adenocarcinoma, and basal cell carcinoma.

A 22-year-old woman sustained ocular laceration complicated by extensive an eyelid degloving injury when she accidentally fell and the drinking glass in her hand broke and cut her in the right eye area. She immediately noticed significant vision loss; thus, early surgical intervention was performed in collaboration with ophthalmologists and plastic surgeons. At 16 months after the injury, a good visual prognosis was achieved (visual acuity >20/40: IOL) without eye movement disorder or eyelid opening disorder, and phthisis bulbi and the need for prosthetic eye wear were avoided. Functional and cosmetic reconstruction can be achieved with concurrent emergency surgical intervention by ophthalmologists and plastic surgeons, even in cases of ocular laceration complicated by extensive eyelid degloving injury, where blindness and phthisis bulbi are predicted.

Introduction: Giant congenital melanocytic nevus (GCMN) is a large melanocytic nevus, and its full-thickness removal is usually difficult due to the lack of skin available for reconstruction. Curettage is an alternative approach in cases of GCMN to remove the superficial dermis above the cleavage plane with a curette in the neonatal period, and its major complications include repigmentation, retarded epithelization, and hypertrophic scar formation. In Japan, the JACE® cultured epidermal autograft (CEA) was approved and covered by public healthcare insurance for the treatment of congenital melanocytic nevus (CMN) that is difficult to treat with conventional methods in 2016. We have used CEA for wounds after curettage in the neonatal period or following ablation after the neonatal period in combination with laser therapies to reduce the above-mentioned complications. Methods: In this study, we summarized all consecutive CMN patients treated using CEA from December 2016 to April 2019 and evaluated the duration required for epithelialization, incidence of hypertrophic scar, and color change in the target nevus by comparing the L∗ values one year later between the Curettage group, the non-Curettage group with initial treatment or the subsequent group. Results: No significant differences were seen in the epithelization period or incidence of hypertrophic scars among the groups, but the color of the target nevus was improved significantly in the Curettage group (p

A 48-year-old man sustained a severe punching hole injury of 8 cm in diameter on the palm of his left hand using a punching instrument (for making the bottom of drink cans), and his middle and ring fingers were almost amputated. A flow-through type immediate ALT (anterolateral thigh) flap was used to bypass the distal blood flow and a titanium plate for mandibular reconstruction and plantar glabrous skin were used to preserve the floating amputated fingers. As there has been no report of reconstruction of a punching hole in the palmar region that exceeds 8 cm in diameter, this report is novel and educational.

Abstract We previously showed that high hydrostatic pressure (HHP) treatment at 200 MPa for 10 min induced complete cell death in skin and skin tumors via necrosis. We used this technique to treat a giant congenital melanocytic nevus and reused the inactivated nevus tissue as a dermis autograft. However, skin inactivated by HHP promoted inflammation in a preclinical study using a porcine model. Therefore, in the present study, we explored the pressurization conditions that induce apoptosis of the skin, as apoptotic cells are not believed to promote inflammation, so the engraftment of inactivated skin should be improved. Using a human dermal fibroblast cell line in suspension culture, we found that HHP at 50 MPa for ≥ 36 h completely induced fibroblast cell death via apoptosis based on the morphological changes in transmission electron microscopy, reactive oxygen species elevation, caspase activation and phosphatidylserine membrane translocation. Furthermore, immunohistochemistry with terminal deoxynucleotidyl transferase dUTP nick-end labeling and cleaved caspase-3 showed most cells in the skin inactivated by pressurization to be apoptotic. Consequently, in vivo grafting of apoptosis-induced inactivated skin resulted in successful engraftment and greater dermal cellular density and macrophage infiltration than our existing method. Our finding supports an alternative approach to hydrostatic pressure application.

Abstract Background Human adipose-derived stem cells (hASCs) are a subset of mesenchymal stem cells (MSCs); it has been regarded as one of the most promising stem cells. We previously found that fibroblast growth factor-2 (FGF-2) enhanced the proliferation and differentiation of hASC. However, the mechanisms involved in the growth of hASCs by FGF-2 have not been investigated. Methods Human adipose-derived stem cells (hASCs) were cultured with FGF-2, and cell growth was assessed. Effects of FGF Receptor (FGFR) inhibitor (NVP-BGJ398), ERK1/2 inhibitor (PD98059), PI3K/Akt inhibitor (LY294002), JNK inhibitor (SP600125), and p38 MAPK inhibitor (SB203580) and Src inhibitor (PP1) on the proliferation were investigated. At the same time, we assessed the effect of FGFR inhibitor on several signaling enzymes such as ERK1/2, JNK, p38, and Akt, in protein level. The involvement of Src activation by FGF-2 was also examined. Results FGF-2 markedly promoted proliferation of hASCs at concentrations lower than 10 ng/ml and stimulated cell progression to the S and G2/M phases. Proliferation was blocked by the FGFR inhibitor (NVP-BGJ398) and various signaling pathway inhibitors, such as Erk1/2 inhibitor (PD98059), PI3K/Akt inhibitor (LY294002), JNK inhibitor (SP600125), and p38MAPK inhibitor (SB203580). The FGFR inhibitor reduced the activation of protein kinases, such as AKT, Erk1/2, JNK, and p38, in several signaling pathways. The downstream kinase of FGFR, Src, was activated by FGF-2, and its activation was canceled by the FGFR inhibitor. MEK1/2, a downstream kinase of Src, was parallelly regulated by FGF-2. The Src inhibitor (PP1) markedly blocked the proliferation of hASCs via inhibition of Src and MEK1/2. Conclusion Src activation is indispensable for FGF-2-mediated proliferation of ASCs, as well as the subsequent activation of multi-signaling pathways.

Introduction: Human platelet lysate (hPL) part of the growth factor cocktail derived from human platelets, which has been applied as a cell growth supplement. The production process is easier in comparison to platelet-rich plasma; thus, hPL is now considered for use in wound healing therapy. However, methods for preserving hPL for more than several months that maintain its bioactivity must be considered, especially for chronic wound treatment. The present study compared the effects of preservation for 9 months using a refrigerator or deep freezer. Methods: We investigated three preservation conditions. In the C-hPL group, hPL was stored at −80 °C in a deep freezer for 9 months; in the CL-hPL group, hPL was cryopreserved for 9 months at −80 °C in a deep freezer then lyophilized; in the L-hPL group, lyophilized hPL was refrigerated at 4 °C for 9 months. The quantity and quality of growth factors in these three groups were measured by an ELISA and in fibroblast cell cultures. Then, gelatin hydrogel discs were impregnated with hPL and its effects with regard to the promotion of wound healing in mice were evaluated by histologic examinations. Results: The PDGF-BB concentration in C-hPL, CL-hPL and L-hPL was 18,363 ± 370 pg/ml, 11,325 ± 171 pg/ml, and 12,307 ± 348 pg/ml, respectively; the VEGF concentration was 655 ± 23 pg/ml, 454 ± 27 pg/ml, and 499 ± 23 pg/ml, respectively; and the TGF-β1 concentration was 97,363 ± 5418 pg/ml, 73,198 ± 2442 pg/ml, and 78,034 ± 3885 pg/ml, respectively. In cell culture medium, fibroblast cell cultures were better supported in the hPL groups than in the fetal bovine serum group. In the histologic examination of the wound healing process, no differences were observed among the three preserved hPL groups with regard to epithelialization, or granulation tissue or capillary formation. The wounds in all groups had almost healed by day 14. Conclusions: The stability of growth factors contained in lyophilized hPL is maintained at 4 °C for up to 9 months. This was a versatile preservation method that can be applied in clinical practice. Keywords: Platelet lysate, Gelatin hydrogel, Cryopreservation, Lyophilization, Long-term preservation, Wound healing

Abstract Background Platelet-rich plasma (PRP) is an autologous blood product that contains a high concentration of several growth factors. Platelet-derived growth factor (PDGF)-BB is a potential mitogen for human adipose-derived stem cells (hASCs). PRP stimulates proliferation of hASCs; however, the signaling pathways activated by PRP remain unclear. Methods hASCs were cultured with or without PRP or PDGF-BB, and proliferation was assessed. hASCs were also treated with PRP or PDGF-BB with or without imatinib, which is a PDGF receptor tyrosine kinase inhibitor, or sorafenib, which is a multikinase inhibitor. Inhibition of cell proliferation was examined using anti-PDGF antibody (Abcam, Cambridge, UK), by cell counting. We assessed the effects of inhibitors of various protein kinases such as ERK1/2, JNK, p38, and Akt on the proliferation of hASCs. Results The proliferation was remarkably promoted in cells treated with either 1% PRP or 10 ng/ml PDGF-BB, and both imatinib and sorafenib inhibited this proliferation. Anti-PDGF antibody (0.5 and 2 μg/ml) significantly decreased the proliferation of hASCs compared with control. PRP-mediated hASC proliferation was blocked by inhibitors of ERK1/2, Akt, and JNK, but not by an inhibitor of p38. Conclusions PRP promotes hASC proliferation, and PDGF-BB in PRP plays a major role in inducing the proliferation of hASCs. PRP promotes hASC proliferation via ERK1/2, PI3K/Akt, and JNK signaling pathways.

Raman spectroscopy can be used for analysis of objects by detecting the vibrational spectrum using label-free methods. This imaging method was applied to analysis of peripheral nerve regeneration by examining the sciatic nerve in vitro and in vivo. Raman spectra of intact nerve tissue had three particularly important peaks in the range 2800 - 3000 cm-1. Spectra of injured sciatic nerves showed significant changes in the ratio of these peaks. Analysis of cellular spectra suggested that the spectrum for sciatic nerve tissue reflects the axon and myelin components of this tissue. Immunohistochemical analysis showed that the number of axons and the myelinated area were reduced at 7 days after injury and then increased by 28 days. The relative change in the axon to myelin ratio showed a similar initial increase, followed by a decrease at 28 days after injury. These changes correlated with the band intensity ratio and the changes in distribution of axon and myelin in Raman spectral analysis. Thus, our results suggest that label-free biochemical imaging with Raman spectroscopy can be used to detect turnover of axon and myelin in peripheral nerve regeneration. [ABSTRACT FROM AUTHOR]

In general, internal plate fixation is mainly performed in order to reduce multiple facial bone fractures and rarely uses an external fixator. Although it would be ideal to successfully achieve or preserve the occlusal relationship of the upper and lower jaws, it is difficult to successfully manage multiple bone fragments in order to use them to achieve internal fixation, especially when treating a severely comminuted facial fracture. In addition, it is also important to prevent the onset of severe infection after sub-periosteal dissection, due to the presence of avascular bony fragments and an internal fixation implant which thus represents a foreign body. In order to treat extensive comminuted fracture without internal plate fixation, we performed external fixation using an Ilizarov-type external fixator. The Ilizarov-type external fixator is characterized by a few circular external structures and threaded rods. This device can be modified for use in the field of orthopedic surgery to correct three-dimensional deformities caused by fragile complicated fractures and bone defects. The advantages of the Ilizarov-type external fixator for treating panfacial fractures include a low invasiveness, utility in various fracture situations, easy adjustment of threaded rods, and a low potential cost. We herein report the first case of a severe panfacial fracture that was successfully treated using an Ilizarov-type external fixator. Keywords: Panfacial fracture, Comminuted facial fracture, Implant infection, Ilizarov-type external fixator, External fixation devices, Bone extending devices

We developed a novel skin regeneration therapy combining nevus tissue inactivated by high hydrostatic pressure (HHP) in the reconstruction of the dermis with a cultured epidermal autograft (CEA). The issue with this treatment is the unstable survival of CEA on the inactivated dermis. In this study, we applied collagen/gelatin sponge (CGS), which can sustain the release of basic fibroblast growth factor (bFGF), to the inactivated skin in order to accelerate angiogenesis. Murine skin grafts from C57BL6J/Jcl mice (8 mm in diameter) were prepared, inactivated by HHP and cryopreserved. One month later, the grafts were transplanted subcutaneously onto the back of other mice and covered by CGS impregnated with saline or bFGF. Grafts were harvested after one, two and eight weeks, at which point the engraftment was evaluated through the histology and angiogenesis-related gene expressions were determined by real-time polymerase chain reaction. Histological sections showed that the dermal cellular density and newly formed capillaries in the bFGF group were significantly higher than in the control group. The relative expression of FGF-2, PDGF-A and VEGF-A genes in the bFGF group was significantly higher than in the control group at Week 1. This study suggested that the angiogenesis into grafts was accelerated, which might improve the engraftment of inactivated dermis in combination with the sustained release of bFGF by CGSs.

Summary:. Nevus lipomatosus cutaneous superficialis is a benign hamartoma characterized by the presence of mature ectopic adipocytes in the dermis. Here, we report a case of giant nevus lipomatosus cutaneous superficialis, measuring 24 × 14 cm in diameter, on the buttock. The tumor was resected, and the cutaneous defect was closed with a rotation flap under general anesthesia. No complication or recurrence occurred after surgery. Giant nevus lipomatosus cutaneous superficialis is very rare, and repair of the skin defect required plastic and reconstructive surgery.

Summary:. For medial canthal tendon reattachment, many kinds of surgical technique have previously been reported. They are complicated and generally require technical skill and prolonged operating times. We, for the first time, describe a novel device, the Caraji Anchor Suture System, used for reattaching the medial canthal tendon to the medial orbital wall after removal of squamous cell carcinoma. The medial canthal tendon was excellently fixed with this rapid and simple method. No complications, such as local infection or exposure, have occurred up to the present. This technique appears to be safe, fast, and effective for reconstruction of the medial canthus after tendon avulsion or loss from excision of cutaneous carcinoma in plastic and reconstructive surgery.

Summary:. Cultured epithelial autograft (CEA) is an epithelial sheet prepared from a patient’s own skin using cell culture. In Japan, CEA (JACE; Japan Tissue Engineering Co., Ltd., Gamagori, Japan) was approved and covered by public health care insurance for use in the treatment of giant congenital melanocytic nevus (GCMN) in 2016. There are several treatment options for GCMN; however, the complete removal of a GCMN is usually difficult due to the lack of skin. In this report, we describe a case of GCMN in the occipital region that was treated using CEA after curettage. A 2-month-old boy had a GCMN of 13 × 21 cm in his occipital region. We used full-thickness skin taken from the back of the right auricle to prepare CEA under general anesthesia at 4 months of age. Three weeks after preparing CEA, we performed curettage of the right half of the GCMN, and CEA was grafted onto the wound afterward. CEA took completely, and epithelization was observed at 10 days after surgery. We then performed curettage with subsequent grafting of CEA on the left half of the GCMN at 7 months of age. CEA took completely, and epithelization was observed in this procedure as well with no hair loss at 8 months of age. Whether or not curettage can reduce the risk of malignant transformation into malignant melanoma of GCMN is unclear; however, the application of CEA after curettage may be a promising option for obtaining early epithelization.

Summary:. We repaired a long horizontal defect in the lower lip caused by the resection of squamous cell carcinoma by reconstructing the white lip with a V-Y advancement flap and vermilion with a tongue flap. During this procedure, we deepithelialized the V-Y flap, lifted the upper margin of the flap and sutured it to the remaining upper margin of the lip, and then covered it with the tongue flap, resulting in the successful reconstruction of a thick lip. There were no postoperative complications in articulation or swallowing. This novel procedure is a simple method to aesthetically and functionally reconstruct a thick lower lip.