Seasonal changes in hydration in free-living Japanese children and adolescents.

Changes in hydration status occur throughout the day affecting physiological and behavioral functions. However, little is known about the hydration status of free-living Japanese children and the seasonality of this response. We evaluated hydration status estimated by urine osmolality (Uosm) in 349 children (189 boys and 160 girls, 9.5 ± 2.6 years, range: 6 to 15 years) upon waking at home and during a single school day in spring (April) and summer (July). Further, we assessed the efficacy of employing self-assessment of urine color (UC, based on an 8-point scale) by children to monitor their hydration status. Early morning Uosm was greater in the spring (903 ± 220 mOsm L-1; n = 326) as compared to summer (800 ± 244 mOsm L-1; n = 125) (P = 0.003, paired t-test, n = 104). No differences, however, were observed in Uosm during the school day (P = 0.417, paired t-test, n = 32). While 66% and 50% of children were considered underhydrated (Uosm ≧800 mOsm L-1) upon waking in the spring and summer periods respectively, more children were underhydrated (∼12%) during the school day. Self-reported UC was similar between seasons as assessed in the morning and school day (P ≧ 0.101, paired t-test), which differed from the pattern of responses observed with Uosm. We showed that a significant number of Japanese children are likely underhydrated especially in the spring period. Children do not detect seasonal changes in hydration from self-assessed UC, limiting its utility to manage hydration status in children.

Elaborate cooperation of poly(rC)-binding proteins 1/2 and glutathione in ferroptosis induced by plasma-activated Ringer's lactate.

Reactive species are involved in various aspects of neoplastic diseases, including carcinogenesis, cancer-specific metabolism and therapeutics. Non-thermal plasma (NTP) can directly provide reactive species, by integrating atmospheric and interjacent molecules as substrates, to represent a handy strategy to load oxidative stress in situ. NTP causes apoptosis and/or ferroptosis specifically in cancer cells of various types. Plasma-activated Ringer's lactate (PAL) is another modality at the preclinical stage as cancer therapeutics, based on more stable reactive species. PAL specifically kills malignant mesothelioma (MM) cells, employing lysosomal ·NO as a switch from autophagy to ferroptosis. However, the entire molecular mechanisms have not been elucidated yet. Here we studied cytosolic iron regulations in MM and other cancer cells in response to PAL exposure. We discovered that cells with higher catalytic Fe(II) are more susceptible to PAL-induced ferroptosis. PAL caused a cytosolic catalytic Fe(II)-associated pathology through iron chaperones, poly (rC)-binding proteins (PCBP)1/2, inducing a disturbance in glutathione-regulated iron homeostasis. PCBP1/NCOA4-mediated ferritinophagy started at a later phase, further increasing cytosolic catalytic Fe(II), ending in ferroptosis. In contrast, PCBP2 after PAL exposure contributed to iron loading to mitochondria, leading to mitochondrial dysfunction. Therapeutic effect of PAL was successfully applied to an orthotopic MM xenograft model in mice. In conclusion, PAL can selectively sensitize MM cells to ferroptosis by remodeling cytoplasmic iron homeostasis, where glutathione and PCBPs play distinct roles, resulting in lethal ferritinophagy and mitochondrial dysfunction. Our findings indicate the clinical application of PAL as a ferroptosis-inducer and the potential of PCBPs as novel targets in cancer therapeutics.

Perceptions of family medicine among long-term patients of a family medicine clinic in Japan: a mixed-methods study.

OBJECTIVES: To examine the perceptions of family medicine among patients attending a family medicine clinic for over 10 years in Japan and explore the unique characteristics of a family medicine which was developed in the local community in northern Japan. DESIGN: Explanatory, sequential mixed-method design comprising a survey by questionnaires and semistructured interviews. SETTING: The study was conducted at one of the oldest family medicine primary care clinics in Japan. We surveyed and interviewed participants from November 2019 to March 2020. PARTICIPANTS: 144 patients who have attended a family medicine clinic since April 2009 completed the questionnaires. Semistructured interviews with nine participants were conducted. RESULTS: Among the respondents, 131 (91%) reported having attended a family medicine clinic. In terms of their perceptions of what 'family physicians' do, 42 (35%) stated 'a doctor who treats various diseases with a general view', 34 (29%) stated 'a doctor who treats outpatients and visit patients' houses', and 15 (13%) stated 'a doctor whom one can consult for anything and is familiar with one's family and lifestyle'. The results of the qualitative analysis revealed two themes with regard to patients' perceptions of family medicine: 'seeing the whole person and referring suitably' and 'medical care at home'. Patients' perceptions of family medicine identified in the quantitative study were strongly associated with the characteristics extracted from the qualitative study. CONCLUSION: Patients attending the family medicine clinic had clear perceptions of what family physicians do. The two major perceptions of the characteristics of family medicine were identified as 'seeing the whole person and referring suitably' and 'medical care at home'.

Ferroptosis induced by plasma-activated Ringer's lactate solution prevents oral cancer progression.

OBJECTIVE: This study aimed to investigate the effect of plasma-activated Ringer's lactate solution (PAL) on oral squamous cell carcinoma (OSCC) cells and carcinogenic processes with a particular focus on iron and collagenous matrix formation. MATERIALS AND METHODS: We used three OSCC cell lines, one keratinocyte cell line, and two fibroblast lines, and cell viability assays, immunoblotting, flow cytometry, and transmission electron microscopy were performed to evaluate the effect and type of cell death. The effect of PAL treatment on lysyl oxidase (LOX) expression was investigated in vitro and in vivo. Tamoxifen-inducible Mob1a/b double-knockout mice were used for the in vivo experiment. RESULTS: PAL killed OSCC cells more effectively than the control nontumorous cells and suppressed cell migration and invasion. Ferroptosis occurred and the protein level of LOX was downregulated in cancer cells in vitro and in vivo. Additionally, PAL improved the survival rate of mice and suppressed collagenous matrix formation. CONCLUSIONS: We demonstrated that PAL specifically kills OSCC cells and that ferroptosis occurs in vitro and in vivo. Furthermore, PAL can prevent carcinogenesis and improve the survival rate of oral cancer, especially tongue cancer, by changing collagenous matrix formation via LOX suppression.

Morphology of the Radial Attachment of the Radioulnar Ligament and Surrounding Landmarks.

PURPOSE: The triangular fibrocartilage complex (TFCC) is composed of dorsal and palmar radioulnar ligaments (RULs). A common injury pattern of the RUL is the avulsion of the ulnar insertion, which can be treated by arthroscopic or open repair. Although the general method of TFCC reconstruction is a tendon graft with a bone tunnel, detailed information regarding the radial attachment of the RUL is unclear. This study aimed to clarify the morphology of the radial attachments of the palmar radioulnar ligament (PRUL), dorsal radioulnar ligament (DRUL), and short radiolunate ligament (SRL) using three-dimensional imaging. METHODS: A total of 29 upper limbs (16 formalin-fixed and 13 embalmed by Thiel's embalming method) of Japanese cadavers were used. After gross observation, we marked the attachments of the PRUL, DRUL, and SRL using 0.7-mm diameter pins. We created three-dimensional images of the radius, outlining the PRUL, DRUL, and SRL attachments. The software application calculated the centers of the PRUL and DRUL attachments. RESULTS: The PRUL attachment was horizontally shaped. The center of the PRUL was 1.5 mm proximal and 5.8 mm radial to the tip of the palmar pyramid formed by the palmar cortex and the radioulnar and radiocarpal joint surfaces. The DRUL attachment was vertically shaped. The center of the DRUL was 2.0 mm proximal and 1.7 mm radial from the tip of the dorsal pyramid formed by the dorsal cortex and the radioulnar and radiocarpal joint surfaces. The length of the SRL was 9.2 mm. The SRL and PRUL were strongly conjoined. CONCLUSIONS: The anatomical center on the RUL attachment of the radius can be determined from osseous landmarks. CLINICAL RELEVANCE: The findings of this study contribute to the understanding of RUL attachment to the distal radius and may assist surgeons in performing anatomical reconstruction of TFCC.

Hypomagnesemia because of nedaplatin for cervical cancer: A case report.

A 76-year-old woman with cervical cancer was treated with nedaplatin, a platinum-based drug. After the initiation of the treatment, she became aware of numbness, dizziness, and loss of appetite. Exploration of the causes revealed no clues, but blood tests revealed hypocalcemia and hypomagnesemia. She was treated with intravenous calcium and magnesium, which resolved calcium, magnesium levels, and her symptoms. She was diagnosed with hypomagnesemia because of nedaplatin. Regular follow-up is necessary for patients during or after nedaplatin. Awareness of electrolyte disturbances may elucidate the accurate diagnosis even in patients with obscure symptoms, particular in undergoing or following anticancer therapies.

Flexor tendon transfer for focal dystonia with involuntary flexion of the ring finger.

We report a case of task-specific dystonia characterized by involuntary flexion of the ring finger. After resecting the flexor of the ring finger, the flexor digitorum profundus of the middle finger was transferred to the resected site.

Hyperthermia maintains death receptor expression and promotes TRAIL-induced apoptosis.

BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand activates apoptotic pathways and could potentially be used in anticancer treatments. However, oral squamous cell carcinoma cells are known to be resistant to tumor necrosis factor-related apoptosis-inducing ligand-induced cell death. It has been previously reported that hyperthermia upregulates tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in other cancers. As such, we evaluated whether hyperthermia upregulates tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in a tumor necrosis factor-related apoptosis-inducing ligand-resistant oral squamous cell carcinoma cell line. METHODS: The oral squamous cell carcinoma cell line HSC3 was cultured and divided into hyperthermia and control groups. We investigated the antitumor effects of recombinant human tumor necrosis factor-related apoptosis-inducing ligand using cell proliferation and apoptosis assays. Additionally, we measured death receptor 4 and 5 levels, and determined death receptor ubiquitination status, as well as E3 ubiquitin ligase targeting of death receptor in both hyperthermia and control groups before recombinant human tumor necrosis factor-related apoptosis-inducing ligand administration. RESULTS: Treatment with recombinant human tumor necrosis factor-related apoptosis-inducing ligand produced greater inhibitory effects in the hyperthermia group than in the control group. Moreover, death receptor protein expression in the hyperthermia group was upregulated on the cell surface (and overall), although death receptor mRNA was downregulated. The half-life of death receptor was several hours longer in the hyperthermia group; concomitantly, E3 ubiquitin ligase expression and death receptor ubiquitination were downregulated in this group. CONCLUSION: Our findings suggested that hyperthermia enhances apoptotic signaling by tumor necrosis factor-related apoptosis-inducing ligand via the suppression of death receptor ubiquitination, which upregulates death receptor expression. These data suggest that the combination of hyperthermia and tumor necrosis factor-related apoptosis-inducing ligand has implications in developing a novel treatment strategy for oral squamous cell carcinoma.

Low-temperature plasma as magic wand to differentiate between the good and the evil.

Plasma is the fourth physical state of matter, characterized by an ionized gaseous mixture, after solid, liquid, and gas phases, and contains a wide array of components such as ions, electrons, radicals, and ultraviolet ray. Whereas the sun and thunder are typical natural plasma, recent progress in the electronics enabled the generation of body-temperature plasma, designated as low-temperature plasma (LTP) or non-thermal plasma since the 1990s. LTP has attracted the attention of researchers for possible biological and medical applications. All the living species on earth utilize water as essential media for solvents and molecular transport. Thus, biological application of LTP naturally intervenes water whether LTP is exposed directly or indirectly, where plasma-activated lactate (PAL) is a standard, containing H2O2, NO2- and other identified molecules. Electron spin resonance and immunohistochemical studies demonstrated that LTP exposure is a handy method to load local oxidative stress. Cancer cells are characterized by persistent self-replication and high cytosolic catalytic Fe(II). Therefore, both direct exposure of LTP and PAL can provide higher damage to cancer cells in comparison to non-tumorous cells, which has been demonstrated in a variety of cancer types. The cell death mode is either apoptosis or ferroptosis, depending on the cancer-type. Thus, LTP and PAL are expected to work as an additional cancer therapy to the established guideline protocols, especially for use in somatic cavities or surgical margins.

Exposure of low-temperature plasma after vaccination in tongue promotes systemic IgM induction against spike protein of SARS-CoV-2.

COVID-19 has been pandemic since 2020 with persistent generation of new variants. Cellular receptor for SARS-CoV-2 is angiotensin-converting enzyme 2 (ACE2), where transmembrane serine protease-2 (TMPRSS2) is essential for viral internalization. We recently reported abundant expression of ACE2 and TMPRSS2 in the oral cavity of humans and mice. Therefore, oral cavity may work for COVID-19 infection gates. Here we undertook to evaluate whether vaccination in the tongue harbors any merit in comparison to subcutaneous injection. Low-temperature plasma (LTP) is the fourth physical state of matters with ionization above gas but at body temperature. LTP provides complex chemistry, eventually supplying oxidative and/or nitrosative stress on the interface. LTP-associated cellular death has been reported to cause apoptosis and/or ferroptosis. However, there is few data available on immunogenicity retention after LTP exposure. We therefore studied the effect of LTP exposure after the injection of keyhole limpet hemocyanin (KLH) or spike 2 protein of SARS-CoV-2 to the tongue of six-week-old male BALB/c mice, compared to subcutaneous vaccination. Whereas LTP did not change the expression of ACE2 and TMPRSS2 in the tongue, repeated LTP exposure after tongue vaccination significantly promoted systemic and specific IgM production at day 11. In contrast, repeated LTP exposure after subcutaneous vaccination of KLH decreased systemic IgM production. Of note, tongue injection produced significantly higher titer of IgM and IgG in the case of KLH. In conclusion, LTP significantly reinforced humoral immunity by IgM after tongue injection. Vaccination to the tongue can be a novel strategy to acquire immediate immunity.

Intraosseous Schwannoma of the Distal Phalanx.

Intraosseous schwannomas are extremely rare and only a few cases involving the proximal phalanx and metacarpal of the hand have been reported. We report a patient with an intraosseous schwannoma of the distal phalanx. Radiographs showed lytic lesions in the bony cortex and enlarged soft shadows of the distal phalanx. The lesion was hyperintense to fat on T2-weighted magnetic resonance imaging (MRI) and strongly enhanced after gadolinium (Gd) administration. Surgical findings revealed that the tumour had developed from the palmar side of the distal phalanx and the medullary cavity was filled with a yellow tumour. The histological diagnosis was schwannoma. A definitive diagnosis of intraosseous schwannoma using radiography is difficult. In our case, a high signal was observed on Gd-enhanced MRI and histological findings showed areas with a high cellular area. Thus, Gd-enhanced MRI may help in the diagnosis of intraosseous schwannomas of the hand. Level of Evidence: Level V (Therapeutic).

Field-effect transistor antigen/antibody-TMDs sensors for the detection of COVID-19 samples.

We fabricated sensors by modifying the surface of MoS2 and WS2 with COVID-19 antibodies and investigated their characteristics, including stability, reusability, sensitivity, and selectivity. Thiols and disulfanes in antibodies strongly interact with vacant Mo or W sites of MoS2 or WS2, yielding durable devices that are stable for several days in the air or water. More importantly, detachment of the antibodies is suppressed even during the aggressive cleaning process of the devices at pH 3, which allows reusing the same device in several experiments without appreciable loss of sensitivity. Therefore, the nanodevice may be employed in samples of different patients. Further, we found a limit of detection (LOD) of 1 fg ml-1 at room temperature, time responses of 1 second, and selectivity against interferences such as KLH protein or Albumin.

Load Distribution in Dorsally-Angulated Distal Radius Deformity Using Finite Element Analysis.

PURPOSE: The load axis of the carpals is located on the volar side of the normal distal radius. A volar lunate facet fracture (VLFF) is exposed to volar-shearing stress, which can cause volar displacement of the carpus. A previous biomechanical study reported that the load at the scaphoid fossa was located more dorsally and the pressure at the lunate fossa decreased in a dorsally-angulated model. However, the distal radius load distribution for various volar tilts remains unclear. We speculate that if the volar tilt decreases, the load distribution moves dorsally and decreases the stress on the VLFF. Therefore, we analyzed a dorsally-angulated distal radius model to evaluate changes in the load distribution using finite element analysis. METHODS: A 3-dimensional finite element wrist model was developed using computed tomography images. The ligaments were modeled as tension-only spring elements. We considered the intact wrist model for a volar tilt of 15° and created 5 additional models for volar tilts of 10°, 5°, 0°, -5°, and -10°. RESULTS: As the dorsal angulation increased, the stress distribution moved from volar to dorsal and from the lunate fossa toward the scaphoid fossa. The maximum stress on the volar lunate facet was reduced as volar tilt decreased. The maximum stress was higher on the lunate fossa for volar tilts from 15° to 5°. In contrast, the maximum stress was higher on the scaphoid fossa for volar tilts of ≤0°. CONCLUSIONS: Load transmission moved from volar to dorsal and from the lunate fossa to the scaphoid fossa when the volar tilt decreased. Therefore, a decrease in the volar tilt would reduce the load on the VLFF. CLINICAL RELEVANCE: This study provides surgeons accurate knowledge regarding load distribution of the distal radius for various volar tilts that could be helpful in treating patients with VLFFs.

Development of a newly immunoassay specific for mouse presepsin (sCD14-ST).

Presepsin (sCD14-ST) is used as a marker for sepsis diagnosis. The production mechanism of presepsin is unique in that it is produced through phagocytosis of microorganisms. However, some studies have demonstrated that non-infected patients had increased presepsin levels and that presepsin is related to the risk or severity of diseases. This study was designed to describe a sensitive sandwich enzyme-linked immunosorbent assay for mouse presepsin developed to investigate the association of presepsin with diseases. Polyclonal antibodies were generated from peptide-immunized rabbit antiserum. Mouse presepsin standard was prepared using the recombinant method as an Fc-fusion protein. The linear detection range of the method was 4.7-300 pg/mL with a detection limit of 1.4 pg/mL. The assay detected mouse presepsin where mouse soluble CD14 (sCD14) was digested by cathepsin D proteinase and the cross-reactivity of sCD14 was not observed. The normal levels of mouse presepsin and sCD14 were compared; 65.9 ± 21.4 pg/mL and 43.2 ± 7.2 ng/mL were determined, respectively. Moreover, the levels of presepsin and sCD14 were compared with a lipopolysaccharide (LPS)-injected sepsis mouse model. The newly developed analytical method had high specificity to presepsin and is an efficient tool for studying the association between presepsin and diseases.

Mechanical Properties of Ti-Nb-Cu Alloys for Dental Machining Applications.

Titanium has excellent biocompatibility and good corrosion resistance and is extensively used in dental implants and denture bases. However, pure titanium lacks the strength for use in dental prostheses that require relatively high strength. We developed 15 different types of Ti-Nb-Cu alloys and investigated their alloy phases and mechanical properties, including tensile and yield strength, elongation after fracture, and Vickers hardness. The alloy phases of Ti-8%Nb-2%Cu and Ti-13%Nb-2%Cu were α + ß, while those of Ti-5%Nb-5%Cu and Ti-10%Nb-5%Cu were α + Ti2Cu. The tensile strength and hardness of these alloys were significantly higher than those of titanium; however, their elongation was less. In particular, the yield strength of these alloys was more than twice that of titanium. These differences in mechanical properties are attributable to solid-solution strengthening and precipitation strengthening. Other compositions with an alloy phase of α + ß + Ti2Cu or ß + Ti2Cu had high hardness but not high strength. These results suggest that the Ti-8%Nb-2%Cu, Ti-5%Nb-5%Cu, Ti-13%Nb-2%Cu, and Ti-10%Nb-5%Cu alloys can be applied to dental prostheses, which are subject to very high forces from accessories such as long-span bridges, clasps, implant-retained superstructures, and narrow-diameter implants.

Radiographic study of the distal ulna in rheumatoid arthritis with extensor digitorum communis ruptures.

OBJECTIVES: Extensor digitorum communis rupture of the wrist often occurs in patients with rheumatoid arthritis. Early operation is desirable for patients with a high risk of rupture; therefore, rheumatologists should diagnose it during daily examinations. This study aimed to clarify radiographic changes in the distal ulna and related factors associated with extensor digitorum communis rupture in patients with rheumatoid arthritis. METHODS: We analysed plain radiographs of 40 patients with rheumatoid arthritis associated with extensor digitorum communis rupture and 62 healthy controls. We investigated the deformation of the distal ulna, Larsen grades, and radiological parameters such as ulnar variance, ulnar bowing angle, dorsal protrusion, and dorsal bowing angle. RESULTS: The ratios of the ulna head deformation, Larsen grades, ulnar variance, dorsal protrusion, and dorsal bowing angle were significantly larger in the ruptured group than in the control group. Multiple logistic regression analysis revealed that dorsal protrusion and Larsen grades were significantly associated with extensor digitorum communis rupture. CONCLUSIONS: Deformity of the distal ulna is evident in patients with an extensor digitorum communis rupture. Ulnar head deformation, high Larsen grades, and large dorsal protrusion are potential risk factors for extensor digitorum communis rupture.

Association of alcohol intake and female gender with high expression of TMPRSS2 in tongue as potential risk for SARS-CoV-2 infection.

COVID-19 is pandemic since 2020 and further information is necessary on the risk factors associated with the infection of SARS-CoV-2. As an entry mechanism, SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) as receptor and transmembrane serine protease 2 (TMPRSS2) to activate fusion with host plasma membrane. Because dysgeusia is an early symptom of COVID-19, we here studied the expression of ACE2 and TMPRSS2 in the tongue and the associated tissues of mice and humans with immunohistochemistry and immunoblot analysis. ACE2 expression was low in the human tongue but was observed in the squamous epithelium, perineurium, arterial wall, salivary glands as well as taste buds. In contrast, mice showed high expression. In sharp contrast, TMPRSS2 expression was high in all the cells mentioned above in humans but relatively low in mice except for salivary glands. We then performed semi-quantitation of immunohistochemistry data of human ACE2 and TMPRSS2 and analyzed for age, sex, alcohol intake, and smoking habit with logistic regression analysis. We found that alcohol intake and female gender were the significant risk factors for increasing TMPRSS2 expression. In conclusion, TMPRSS2 is an important factor to be considered regarding SARS-CoV-2 entry and amplification in the oral cavity, which is promoted through drinking habit.

Three-dimensional imaging of the distal radius with reference to volar locking plate surgery.

BACKGROUND: The watershed line is widely accepted as the distal limit of the volar locking plate (VLP); however, the VLP placement could vary depending on the plate contour and morphology of the distal radius. The aim of this study was to investigate the morphology of the distal radius and VLP fitting using 3D images. HYPOTHESIS: We hypothesized that attachment of the VLP would affect the contour of the volar prominence of the distal radius. PATIENTS AND METHODS: Variable-angle LCP two-column volar distal radius Plate 2.4 and 16 formalin-fixed cadavers were studied. The plate and forearm were scanned using a computed tomography scanner. The plate was fixed to the radial shaft in the most distal position without flexor pollicis longus tendon contact. Postero-antero and lateral radiographs were obtained using fluoroscopy. Postero-antero radiographs were superimposed on a 3D image of the distal radius. The virtual plate was attached to the distal radius in the computer simulations and the plate was adjusted in the sagittal plane. In the postero-antero radiographs, the distance between the plate and distal end of the radius (DPR) was measured. In the sagittal plane, the height of the volar lunate facet (VLF) and the plate-to-bone distance of each locking screw hole was measured. The volar cortical angle (VCA) was measured as the angle formed by a line drawn along the volar surface and a line drawn on the radial shaft on the sagittal plane at each locking screw plane. RESULTS: A significant correlation was observed between the height of the VLF and the DPR and between the height of the VLF and the VCA. The plate-to-bone distance at the ulnar screw hole was significantly smaller than that of the other screw holes. DISCUSSION: Our study revealed that the higher the VLF, the more proximal is the VLP. The plate fits on the bone surface at the ulnar side, whereas the radial side has more space between the plate and bone. LEVEL OF EVIDENCE: III, diagnostic Level.

Diluted aqueous extract of heat-not-burn tobacco product smoke causes less oxidative damage in fibroblasts than conventional cigarette.

Smoke from conventional cigarettes (C-cigarettes) contains various reactive oxygen species and toxic chemicals, which potentially cause oxidative damage not only to airways but also to the whole body, leading eventually to diseases, including emphysema, advanced atherosclerosis, and cancer. Many heat-not-burn tobacco products (HTPs) have been commercialized recently in Japan to maintain the smoking population by advertising that HTPs are less toxic. However, there were few studies reported from neutral organizations whether HTPs are indeed less damaging. To evaluate the potential capacity of HTPs to induce oxidative stress, we here compared two different HTPs with two types of C-cigarettes, using human fibroblast IMR90SV cells and 5% aqueous extracts in 10-ml phosphate-buffered saline (50-ml smoke/10 s). HTPs exhibited significantly lower oxidative toxicity in comparison to C-cigarettes. Whereas C-cigarettes induced ferroptosis in fibroblasts, the effects of HTPs were significantly reduced by measuring the levels of peroxides, pro-inflammatory cytokine expression, autophagy, catalytic Fe(II) and 8-hydroxy-2'-deoxyguanosine. Notably, major portions of C-cigarettes-induced pathogenic responses were inhibited by catalase. However, HTPs still induced p62 autophagy-adaptor at 5%-dilution and caused lethal effects to fibroblasts with undiluted solution. In conclusion, HTPs smoke per se can be toxic despite less toxicity in comparison to C-cigarettes, which warrants further investigation.