Sirtinol Supresses Trophozoites Proliferation and Encystation of Acanthamoeba via Inhibition of Sirtuin Family Protein

The encystation of Acanthamoeba leads to the development of metabolically inactive and dormant cysts from vegetative trophozoites under unfavorable conditions. These cysts are highly resistant to anti-Acanthamoeba drugs and biocides. Therefore, the inhibition of encystation would be more effective in treating Acanthamoeba infection. In our previous study, a sirtuin family protein—Acanthamoeba silent-information regulator 2-like protein (AcSir2)—was identified, and its expression was discovered to be critical for Acanthamoeba castellanii proliferation and encystation. In this study, to develop Acanthamoeba sirtuin inhibitors, we examine the effects of sirtinol, a sirtuin inhibitor, on trophozoite growth and encystation. Sirtinol inhibited A. castellanii trophozoites proliferation (IC50=61.24 μM). The encystation rate of cells treated with sirtinol significantly decreased to 39.8% (200 μM sirtinol) after 24 hr of incubation compared to controls. In AcSir2-overexpressing cells, the transcriptional level of cyst-specific cysteine protease (CSCP), an Acanthamoeba cysteine protease involved in the encysting process, was 11.6- and 88.6-fold higher at 48 and 72 hr after induction of encystation compared to control. However, sirtinol suppresses CSCP transcription, resulting that the undegraded organelles and large molecules remained in sirtinol-treated cells during encystation. These results indicated that sirtinol sufficiently inhibited trophozoite proliferation and encystation, and can be used to treat Acanthamoeba infections.

The Pilot Study of Robot-Assisted Training for the Lower Extremity Rehabilitation of Burn Patients

Purpose@#Scar contracture influence the outcome of burn patients significantly. This study aims to investigate the feasibility of robot-assisted training for the lower extremity rehabilitation of burn patients. @*Methods@#This pilot study was conducted on 7 burn patients for 8 weeks between January 2019 and November 2019. Two of 7 patients withdrew from this study because one had skin abrasion on the legs which thigh fastening devices were applied on and the other was not participate in the assessment at 4 weeks after training. Final 5 patients received gait training with SUBAR Ⓡ and numeric rating scale (NRS), 6-minutes walking test, and range of motion in flexion and extension of knee and ankle joint were evaluated before training, 4 weeks and 12 weeks after training. @*Results@#The subjects had a mean age of 51.8±98 years, mean total burn surface area of 30.8±13.7%, mean duration from injury to 1 st assessment of 102.8±39.3 days. Anyone of 5 patients did not have musculoskeletal or cardiovascular side effects such as increased or decreased blood pressure or dizziness. The significant improvement in NRS, gait speed, and range of motion in knee extension and ankle plantarflexion after robotic training (all P＜0.05). @*Conclusion@#Robot-assisted training could be feasible for the rehabilitation of burn patients and it could improve muscle strength and range of motion in lower extremities, and gait function.

Wound Healing Potential of Low Temperature Plasma in Human Primary Epidermal Keratinocytes

BACKGROUND: Low temperature plasma (LTP) was recently shown to be potentially useful for biomedical applications such as bleeding cessation, cancer treatment, and wound healing, among others. Keratinocytes are a major cell type that migrates directionally into the wound bed, and their proliferation leads to complete wound closure during the cutaneous repair/regeneration process. However, the beneficial effects of LTP on human keratinocytes have not been well studied. Therefore, we investigated migration, growth factor production, and cytokine secretion in primary human keratinocytes after LTP treatment.METHODS: Primary cultured keratinocytes were obtained from human skin biopsies. Cell viability was measured with the EZ-Cytox cell viability assay, cell migration was evaluated by an in vitro wound healing assay, gene expression was analyzed by quantitative real-time polymerase chain reaction, and protein expression was measured by enzyme-linked immunosorbent assays and western blotting after LTP treatment.RESULTS: Cell migration, the secretion of several cytokines, and gene and protein levels of angiogenic growth factors increased in LTP-treated human keratinocytes without associated cell toxicity. LTP treatment also significantly induced the expression of hypoxia inducible factor-1α (HIF-1α), an upstream regulator of angiogenesis. Further, the inhibition of HIF-1α expression blocked the production of angiogenic growth factors induced by LTP in human keratinocytes.CONCLUSION: Our results suggest that LTP treatment is an effective approach to modulate wound healing-related molecules in epidermal keratinocytes and might promote angiogenesis, leading to improved wound healing.

The Application of Three-Dimensional Printed Finger Splints for Post Hand Burn Patients: A Case Series Investigation

The application of three-dimensional (3D) printing is growing explosively in the medical field, and is especially widespread in the clinical use of fabricating upper limb orthosis and prosthesis. Advantages of 3D-printed orthosis compared to conventional ones include its lower cost, easier modification, and faster fabrication. Hands are the most common body parts involved with burn victims and one of the main complications of hand burns are finger joint contractures. Applying orthotic devices such as finger splints are a well-established essential element of burn care. In spite of the rapid evolution of the clinical use of 3D printing, to our knowledge, its application to hand burn patients has not yet been reported. In this study, the authors present a series of patients with hand burn injuries whose orthotic needs were fulfilled with the application of 3D-printed finger splints.

Molecular and Biochemical Properties of a Cysteine Protease of Acanthamoeba castellanii

Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

Diversity of vir Genes in Plasmodium vivax from Endemic Regions in the Republic of Korea: an Initial Evaluation

Variant surface antigens (VSAs) encoded by pir families are considered to be the key proteins used by many Plasmodium spp. to escape the host immune system by antigenic variation. This attribute of VSAs is a critical issue in the development of a novel vaccine. In this regard, a population genetic study of vir genes from Plasmodium vivax was performed in the Republic of Korea (ROK). Eighty-five venous blood samples and 4 of the vir genes, namely vir 27, vir 21, vir 12, and vir 4, were selected for study. The number of segregating sites (S), number of haplotypes (H), haplotype diversity (Hd), DNA diversity (π and Θw), and Tajima’s D test value were conducted. Phylogenetic trees of each gene were constructed. The vir 21 (S=143, H=22, Hd=0.827) was the most genetically diverse gene, and the vir 4 (S=6, H=4, Hd=0.556) was the opposite one. Tajima’s D values for vir 27 (1.08530, P>0.1), vir 12 (2.89007, P0.1) were positive, and that of vir 4 (−1.32162, P>0.1) was negative. All phylogenetic trees showed 2 clades with no particular branching according to the geographical differences and cluster. This study is the first survey on the vir genes in ROK, providing information on the genetic level. The sample sequences from vir 4 showed a clear difference to the Sal-1 reference gene sequence, whereas they were very similar to those from Indian isolates.

Extracorporeal Shock Wave Therapy For Treatment of Intractable Stump Pain

Amputation stump pains can be developed in amputation sites after high voltage electrical burn injuries. We experienced one case of these severe stump pains in an upper extremity amputation patient. A 35-year-old man had a 38% total body surface area high voltage electrical burn. The patient underwent skin grafting and left shoulder disarticulation. During the rehabilitation period, he complained about severe stump area pains and phantom pains. We injected 0.5% Bupivacaine and Triamcinolone on the stump neuroma site but the pain sustained. After extracorporeal shock wave therapy (ESWT,) the pain subsided and did not recur. The patient was satisfied with functional and pain outcomes, so we report this case.

Prevalence of Trichomonas vaginalis in Women Visiting 2 Obstetrics and Gynecology Clinics in Daegu, South Korea

This study explored epidemiological trends in trichomoniasis in Daegu, South Korea. Wet mount microscopy, PCR, and multiplex PCR were used to test for Trichomonas vaginalis in vaginal swab samples obtained from 621 women visiting 2 clinics in Daegu. Of the 621 women tested, microscopy detected T. vaginalis in 4 (0.6%) patients, PCR detected T. vaginalis in 19 (3.0%) patients, and multiplex PCR detected T. vaginalis in 12 (1.9%) patients. Testing via PCR demonstrated high sensitivity and high negative predictive value for T. vaginalis. Among the 19 women who tested positive for T. vaginalis according to PCR, 94.7% (18/19) reported vaginal signs and symptoms. Notably, more than 50% of T. vaginalis infections occurred in females younger than 30 years old, and 58% were unmarried. Multiplex PCR, which simultaneously detects pathogens from various sexually transmitted infections, revealed that 91.7% (11/12) of patients were infected with 2 or more pathogens. Mycoplasma hominis was the most prevalent co-infection pathogen with T. vaginalis, followed by Ureaplasma urealyticum and Chlamydia trachomatis. Our results indicate that PCR and multiplex PCR are the most sensitive tools for T. vaginalis diagnosis, rather than microscopy which has been routinely used to detect T. vaginalis infections in South Korea. Therefore, clinicians should take note of the high prevalence of T. vaginalis infections among adolescent and young women in order to prevent persistent infection and transmission of this disease.

Effects of a Modified Hand Compression Bandage for Treatment of Post-Burn Hand Edemas

OBJECTIVE: To evaluate the effect of a modified hand compression bandage in patients with a post-burn hand edema. METHODS: Patients were recruited from burn centers. We classified the patients into two groups: the modified hand compression bandage group comprising of 22 patients who had a modified hand compression bandage and received conventional physical therapy and the conventionally treated group, comprising of 20 patients who received only conventional physical therapy during the 4-week period post-burn. Hand circumference, hand skin thickness, and hand function were evaluated by grip strength, active range of motion (ROM), Jebsen hand function test, and visual analogue scale (VAS). These assessments were used to evaluate treatment effectiveness prior to the first treatment, 2 weeks after the first treatment, 4 weeks after the first treatment, and 4 months after the first treatment. RESULTS: As a result of repeated-measures analysis of variance on hand circumference, skin thickness, VAS, and each metacarpophalangeal joint ROM, we found significant differences that corresponded to time effect (p0.05). CONCLUSION: The modified hand compression bandage will be clinically useful for the treatment of patients with post-burn hand edemas.

Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

Development of Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Azumiobodo hoyamushi (Kinetoplastea)

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

Mirror Writing after Thalamic Hemorrhagic Stroke: A Case Report

Mirror-writing is the process of reversing individual letter and composing word sequence in opposite direction. Unintentional mirror writing has been observed in young children learning to write and interpreted as the manifestation of different cognitive impairments. It is very rare that normal right-handed adult shows mirror writing. However, the acquired "mirror writing" may be shown in left hemispheric stroke, neurodegenerative disease, and diffuse cerebral disorders. To explain this event, some assumption have been indicated such as the motor, the visual dominance, the supplementary motor area, the visio-spatial, the visual word-form, the hemispatial factor or directional and the reflected graphemic representation hypotheses. It is reported that the lesions which causes the "Mirror-writing" are the parietal lobe, basal ganglia, thalamus and right supplementary motor area, etc. We reported a case of "mirror-writing" with left thalamic hemorrhagic stroke.

Epigallocatechin-3-gallate Inhibits the Expression of Adhesion Molecules by Blocking Nuclear Factor Kappa B Signaling in Intestinal Epithelial Cells

BACKGROUND/AIMS: Epigallocatechin-3-gallate (EGCG) is the main polyphenol in green tea and has anti-inflammatory and anti-oxidative effects. The aim of this study was to determine the impact of EGCG on the expression of adhesion molecules and lipopolysaccharide (LPS)-induced nuclear factor-kappa B (NF-kappaB) signaling in rat intestinal epithelial (RIE) cells. METHODS: The effect of EGCG on LPS-induced NF-kappaB signaling and expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 was examined by reverse transcription polymerase chain reaction, western blotting, immunofluorescence and electrophoretic mobility shift assay. RESULTS: LPS-induced expression of ICAM-1 and VCAM-1 mRNA was inhibited by EGCG treatment in RIE cells. LPS-induced inhibitor of kappa B alpha degradation and NF-kappaB nuclear translocation were blocked by EGCG in RIE cells. EGCG blocked LPS-induced NF-kappaB DNA-binding activity in RIE cells. The pharmacological NF-kappaB inhibitor Bay11-7082 suppressed the LPS-induced expression of ICAM-1 and VCAM-1 mRNA in RIE cells. CONCLUSIONS: These results indicate that EGCG inhibits LPS-induced ICAM-1 and VCAM-1 expression by blocking NF-kappaB signaling in intestinal epithelial cells.

Epigallocatechin-3-gallate Inhibits LPS-Induced NF-kappaB and MAPK Signaling Pathways in Bone Marrow-Derived Macrophages

BACKGROUND/AIMS: Epigallocatechin-3-gallate (EGCG), the primary catechin in green tea, has anti-inflammatory and anti-oxidative properties. The aim of the current study was to characterize the impact of EGCG on lipopolysaccharide (LPS)-induced innate signaling in bone marrow-derived macrophages (BMMs) isolated from ICR mice. METHODS: The effect of EGCG on LPS-induced pro-inflammatory gene expression and nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinase (MAPK) signaling was examined using reverse transcription-polymerase chain reaction, Western blotting, immunofluorescence, and the electrophoretic mobility shift assay. RESULTS: EGCG inhibited accumulation of LPS-induced IL-12p40, IL-6, MCP-1, ICAM-1, and VCAM-1 mRNA in BMMs. EGCG blocked LPS-induced IkappaBalpha degradation and RelA nuclear translocation. EGCG blocked the DNA-binding activity of NF-kappaB. LPS-induced phosphorylation of ERK1/2, JNK, and p38 was inhibited by EGCG. U0126 (an inhibitor of MEK-1/2) suppressed the LPS-induced IL-12p40, IL-6, MCP-1, ICAM-1, and VCAM-1 mRNA accumulation in BMMs. CONCLUSIONS: These results indicate that EGCG may prevent LPS-induced pro-inflammatory gene expression through blocking NF-kappaB and MAPK signaling pathways in BMMs.

Effect of Decannulation on Pharyngeal and Laryngeal Movement in Post-Stroke Tracheostomized Patients

OBJECTIVE: To investigate effects of tracheostomy tube on the movement of the hyoid bone and larynx during swallowing by quantitative analysis of videofluoroscopic swallowing study. METHOD: 19 adult stroke patients with tracheostomies, who met the criteria of decannulation participated. Serial videofluroscopic swallowing studies were done over 14 days before decannulation, within 24 hours before decannulation, within 24 hours after decannulation, and over 14 days after decannulation. The kinematic parameter such as pharyngeal transition time, stage transition duration, maximal hyoid bone movement, and maximal laryngeal prominence movement were obtained by 2-D quantitative analysis of videofluoroscopic swallowing study. RESULTS: Pharyngeal transition time and stage transition duration were not significantly changed all the time. The maximal hyoid bone movement and maximal laryngeal prominence just after decannulation were improved significantly compared to just before decannulation (p<0.05), especially on vertical movement. CONCLUSION: The hypothesis that a tracheostomy tube disturbs the hyoid bone and laryngeal movement during swallowing may be supported by this study.

Obturator Prosthesis for Velopharyngeal Insufficiency after Treatment of Soft Palate Cancer: A Case Report

Velopharyngeal insufficiency after surgical resection of soft palate cancer can be troublesome. This report concerns a male suffered from severe dysphagia following combined treatment for soft palate cancer. Sequential videofluoroscopic swallowing studies (VFSS) were used to assess his swallowing function and plan the interventional strategies. Initial VFSS showed huge nasal regurgitation, increased oral transit time, residues in oral cavity, delayed swallowing reflex, pharyngeal residue, impaired laryngeal elevation, and aspiration in semisolid and liquid trials. Obturator prosthesis was fabricated to minimize velopahryngeal insufficiency. After application of obturator prosthesis, swallowing dysfunction in oral and pharyngeal stages was markedly improved. Nasal regurgitation was not shown. Oral residue, oral transit time in oral stage also improved. Residue on vallaculae and pyriform sinuses decreased in pharyngeal stage. Aspiration also decreased. We reported successful obturator prosthesis application with sequential changes of clinical and VFSS findings in our case.

Foot Deformity in Charcot Marie Tooth Disease According to Disease Severity

OBJECTIVE: To investigate the characteristics of foot deformities in patients with Charcot-Marie-Tooth (CMT) disease compared with normal persons according to severity of disease. METHOD: Sixty-two patients with CMT disease were recruited for this study. The normal control group was composed of 28 healthy people without any foot deformity. Patients were classified into a mild group and a moderate group according to the CMT neuropathy score. Ten typical radiological angles representing foot deformities such as pes equinus and pes varus were measured. The CMT group angles were compared with those of the normal control group, and those of the mild group were also compared with those of the moderate group. RESULTS: The lateral (Lat.) talo-first metatarsal angle, anteroposterior talo-first metatarsal angle, Lat. calcaneal-first metatarsal angle, Lat. naviocuboid overlap, Lat. calcaneal pitch, Lat. tibiocalcaneal angle, and Lat. talocalcaneal angle in the CMT group showed a significant difference compared to the normal control group (p<0.05). These findings revealed CMT patients have pes cavus, forefoot adduction, midfoot supination and pes varus deformity. Compared to the mild group, the moderate group significantly showed an increased Lat. calcaneal pitch and decreased Lat. calcaneal-first metatarsal angle, Lat. tibiocalcaneal angle, Lat. talocalcaneal angle, and Lat. talo-first metatarsal angle (p<0.05). These findings revealed that the pes cavus deformity of CMT patients tend to be worse with disease severity. CONCLUSION: The characteristic equinovarus foot deformity patterns in CMT patients were revealed and these deformities tended to be worse with disease severity. Radiographic measures may be useful for the investigation of foot deformities in CMT patients.

Rick Factors Associated with Aspiration in Patients with Head and Neck Cancer

OBJECTIVE: To determine the major risk factors and abnormal videofluoroscopic swallowing study (VFSS) findings associated with aspiration in patients with head and neck cancer (HNC). METHOD: Risk factors associated with aspiration were investigated retrospectively in 241 patients with HNC using medical records and pre-recorded VFSS. Age, gender, lesion location and stage, treatment factors, and swallowing stage abnormalities were included. RESULTS: Aspiration occurred in 50.2% of patients. A univariate analysis revealed that advanced age, increased duration from disease onset to VFSS, higher tumor stage, increased lymph node stage, increased American Joint Committee on Cancer (AJCC) stage, operation history, chemotherapy history, and radiotherapy history were significantly associated with aspiration (p<0.05). Among them, advanced age, increase AJCC stage, operation history, and chemotherapy history were significantly associated with aspiration in the multivariate analysis (p<0.05). Delayed swallowing reflex and reduced elevation of the larynx were significantly associated with aspiration in the multivariate analysis (p<0.05). CONCLUSION: The major risk factors associated with aspiration in patients with HNC were advanced age, higher AJCC stage, operation history, and chemotherapy history. A VFSS to evaluate aspiration is needed in patients with NHC who have these risk factors. Delayed swallowing reflex and reduced elevation of the larynx were major abnormal findings associated with aspiration. Dysphagia rehabilitation should focus on these results.

Neural Substrates of Motor Learning

Motor Learning is a relatively permanent change in the capability for skilled motor performance as a result of practice or experience. Rehabilitation is fundamentally a process of relearning. With advanced neuro-imaging, the roles of brain areas related to motor learning have been revealed. However, results of a single study represent only parts of a puzzle of the neuronal changes underlying motor learning. The neural substrates of motor learning with a combined view of functional imaging data were reviewed.

Assessment of Post-Stroke Cognitive Dysfunction Using 3-Dimensional Virtual Reality Program

OBJECTIVE: To test the feasibility of newly developed 3- dimensional virtual reality (VR) program for assessing the post-stroke cognitive dysfunction. METHOD: Thirty five post-stroke patients and twenty normal healthy subjects were recruited in this study, and post-stroke patients were classified into three groups according to the severity of cognitive dysfunction. We developed three dimensional virtual reality program to assess the cognitive function with virtual subway environment from taking a subway to arriving at one's destination. The total score, number of success and cue were obtained during completing virtual tasks. We investigated the test-retest reliability, and the parameters of the 3-dimensional VR program were compared with Korean Mini-mental status examination. RESULTS: All parameters of the VR program were significantly correlated with MMSE score (p<0.01), and showed the significant difference between patient subgroups and control group (p<0.05). The test-retest reliability of the VR program was significantly high (p<0.01). CONCLUSION: Three dimensional virtual reality program may be helpful to assess the cognitive function in patients with stroke.