Methotrexate-Induced Accelerated Nodulosis in a Patient with Systemic Lupus Erythematosus.

Methotrexate (MTX)-induced accelerated nodulosis (MIAN) reportedly occurs in patients with rheumatic arthritis receiving MTX therapy. However, it has also been reported in patients with other autoinflammatory conditions, such as systemic lupus erythematosus (SLE). A 38-year-old woman diagnosed with SLE presented with multiple movable, firm, flesh-colored nodules on both hands that had developed 3 years ago. She was taking oral medications, specifically hydroxychloroquine, azathioprine, and MTX. Histopathological examination revealed palisaded granulomatous inflammation, surrounded by histiocytes and lymphocytes, along the dermis to the subcutaneous fat layer. Fibrinoid degeneration was observed at the center of the granulomatous inflammation, and dermal mucin deposition was not observed. The patient was diagnosed with MIAN, and therefore discontinuation of MTX was recommended. Subsequently, the lesions almost completely disappeared with no signs of recurrence. MIAN exhibits clinicopathological features similar to those of rheumatoid nodules; therefore, it can be easily misdiagnosed. Herein, we report a case of MIAN in a patient with SLE to contribute to the accurate diagnosis and appropriate management.

Intradermal Low-Fat Spindle Cell Lipoma: A Case Report.

Spindle cell lipoma is a rare benign neoplasm that features a mixture of evenly aligned spindle cells, mature adipocytes, and ropey collagen. Most cases of spindle cell lipoma are found in the subcutaneous tissue, and intradermal spindle cell lipoma is rarely reported. We present a case of intradermal spindle cell lipoma in a 46-year-old female who presented with a 0.7-cm flesh-colored and dome-shaped nodule on the right temple that had developed 6 years ago. This mass was excised, and upon histopathologic examination, an unencapsulated lesion was located in the dermis, which consisted of bland spindle cells, scanty mature adipocytes, rare lipoblasts, and ropey collagen bundles with prominent basophilic myxoid stroma. Immunohistochemical staining showed diffuse positivity for CD34, negativity for the S-100 protein, and loss of retinoblastoma protein expression. Based on these features, intradermal low-fat spindle cell lipoma was diagnosed. No evidence of local recurrence was observed 4 months after excision. Intradermal low-fat spindle cell lipomas are extremely rare and can easily be mistaken for tumors that have similar clinical and histopathological findings. Herein, we report a globally rare case of an intradermal low-fat spindle cell lipoma.

Superficial CD34-Positive Fibroblastic Tumor: Two Case Reports.

Superficial CD34-positive fibroblastic tumor (SCPFT) is a recently described disease entity characterized by marked nuclear pleomorphism, low mitotic count, and diffuse CD34 positivity. It is a rare, distinctive, low-grade fibroblastic neoplasm. To date, only 44 cases have been reported in the English-language literature. Herein, we report two cases of SCPFT involving a 48-year-old male and a 22-year-old male with superficial tumors on the right and left thighs, respectively. Excision was performed in both cases. Histologically, both tumors showed spindle-to-epithelioid cells arranged in fascicular or sheet-like patterns. Most cells displayed granular or eosinophilic glassy cytoplasm, marked nuclear pleomorphism, and a low mitotic rate. On immunohistochemical staining, tumor cells were diffusely positive for CD34 and negative for S100 protein, smooth muscle actin, and desmin. After wide excision, neither patient experienced recurrence or metastasis after 16 months and 11 months of clinical follow-up, respectively. To the best of our knowledge, these are the first two cases of SCPFT reported in Korea. We believe these case reports would contribute to the clinicopathological understanding of SCPFT and assist clinicians in differentiating this tumor from other superficial soft tissue neoplasms.

Efficiency of Ppy-PA-pani and Ppy-PA composite adsorbents in Chromium(VI) removal from aqueous solution.

In this study, first time the combination of composites with Phytic acid (PA) as the organic binder cross-linker is reported. The novel use of PA with single and double conducting polymers (polypyrrole (Ppy) and polyaniline (Pani)) were tested against removal of Cr(VI) from wastewater. Characterizations (FE-SEM, EDX, FTIR, XRD, XPS) were performed to study the morphology and removal mechanism. The adsorption removal capability of Polypyrrole - Phytic Acid - Polyaniline (Ppy-PA-Pani) was deemed to be higher than Polypyrrole - Phytic Acid (Ppy-PA) due to the mere existence of Polyaniline as the extra polymer. The kinetics followed 2nd order with equilibration at 480 min, but Elovich model confirmed that chemisorption is followed. Langmuir isotherm model exhibited maximum adsorption capacity of 222.7-321.49 mg/g for Ppy-PA-Pani and 207.66-271.96 mg/g for Ppy-PA at 298K-318K with R2 values of 0.9934 and 0.9938 respectively. The adsorbents were reusable for 5 cycles of adsorption-desorption. The thermodynamic parameter, ΔH shows positive values confirmed the adsorption process was endothermic. From overall results, the removal mechanism is believed to be chemisorption through Cr(VI) reduction to Cr(III). The use of phytic acid (PA) as organic binder with combination of dual conducting polymer (Ppy-PA-Pani) was invigorating the adsorption efficiency than just single conducting polymer (Ppy-PA).

Experimentally and spectroscopically evidenced mechanistic study of butyl peroxyacid oxidative degradation of benzo[a]pyrene in soil.

Benzo[a]pyrene (BaP) is a major indicator of soil contamination and categorized as a highly persistent, carcinogenic, and mutagenic polycyclic aromatic hydrocarbon. An advanced peroxyacid oxidation process was developed to reduce soil pollution caused by BaP originating from creosote spills from railroad sleepers. The pH, organic matter, particle size distribution of soil, and concentrations of BaP and heavy metals (Cd, Cu, Zn, Pb, and As) in the BaP-contaminated soils were estimated. A batch experiment was conducted to determine the effects of organic acid type, soil particle size, stirring speed, and reaction time on the peroxyacid oxidation of BaP in the soil samples. Additionally, the effect of the organic acid concentration on the peroxyacid degradation of BaP was investigated using an oxidizing agent in spiked soil with and without hydrogen peroxide. The results of the oxidation process indicated that BaP and heavy metal residuals were below acceptable Korean standards. A significant difference in the oxidative degradation of BaP was observed between the spiked and natural soil samples. The formation of a peroxyacid intermediate was primarily responsible for the enhanced BaP oxidation. Further, butyric acid could be reused thrice without losing the efficacy (<90%). The systematic peroxyacid oxidative degradation mechanism of BaP was also discussed. A qualitative analysis of the by-products of the BaP reaction was conducted, and their corresponding toxicities were determined for possible field applications. The findings conclude that the developed peroxyacid oxidation method has potential applications in the treatment of BaP-contaminated soils.

Weight Change Alters the Small RNA Profile of Urinary Extracellular Vesicles in Obesity.

INTRODUCTION: Various kidney diseases reportedly show different urinary extracellular vesicle (EV) RNA profiles. Although obesity is one of the main causes of chronic kidney disease, the expression pattern of urinary EV RNA in obesity is uncertain. Our aim was to sequence the small RNA profiles of urinary EVs in obese patients before and after weight reduction and compare them to those of healthy volunteers (HVs). METHODS: We recruited age-sex-matched obese patients and HVs. The small RNA profiles of urinary EVs were analyzed using RNA sequencing. To evaluate the effect of weight reduction, small RNA profiles of urinary EVs 6 months after bariatric surgery were also analyzed. RESULTS: The proportion of urinary EVs transfer RNA and microRNA of obese patients differed from that of HVs. Obese patients showed differential expression of 1,343 small RNAs in urinary EVs compared to HVs (fold change ≥2 and p value <0.05). Among those, 61 small RNAs were upregulated in obese patients and downregulated after weight reduction, whereas 167 small RNAs were downregulated in obese patients and upregulated after weight reduction. RNA sequencing revealed the correlation between the specific urinary EV small RNAs and clinical parameters including body weight, low-density lipoprotein cholesterol, triglyceride, high-density lipoprotein cholesterol, serum glucose, estimated glomerular filtration rate, and albuminuria. CONCLUSION: Obese patients showed distinct urinary EV small RNA profiles compared to HVs. Weight reduction altered urinary EV small-RNA profiles in obese patients.

The Relationship Between Breast Density Change During Menopause and the Risk of Breast Cancer in Korean Women.

BACKGROUND: The aim of this study was to investigate the relationship between changes in breast density during menopause and breast cancer risk. METHODS: This study was a retrospective, longitudinal cohort study for women over 30 years of age who had undergone breast mammography serially at baseline and postmenopause during regular health checkups at Samsung Medical Center. None of the participants had been diagnosed with breast cancer at baseline. Mammographic breast density was measured using the American College of Radiology Breast Imaging Reporting and Data System. RESULTS: During 18,615 person-years of follow-up (median follow-up 4.8 years; interquartile range 2.8-7.5 years), 45 participants were diagnosed with breast cancer. The prevalence of dense breasts was higher in those who were younger, underweight, had low parity or using contraceptives. The cumulative incidence of breast cancer increased 4 years after menopause in participants, and the consistently extremely dense group had a significantly higher cumulative incidence (CI) of breast cancer compared with other groups [CI of extremely dense vs. others (incidence rate per 100,000 person-years): 375 vs. 203, P < 0.01]. CONCLUSION: Korean women whose breast density was extremely dense before menopause and who maintained this density after menopause were at two-fold greater risk of breast cancer. PREVENTION RELEVANCE: Extremely dense breast density that is maintained persistently from premenopause to postmenopause increases risk of breast cancer two fold in Korean women. Therefore, women having risk factors should receive mammography frequently and if persistently extremely dense breast had been detected, additional modalities of BC screening could be considered.

Prevalence and Characteristics of Onychomycosis in Patients with Knee Osteoarthritis: A Single-centre Prospective Cross-sectional Study.

Knee disorders that compromise patients' lower leg movements and self-care may put these patients at greater risk of onychomycosis. However, little is known about the prevalence of onychomycosis in patients with knee diseases. This study evaluated the prevalence and characteristics of onychomycosis in patients with knee osteoarthritis. A total of 520 consecutive patients with symptomatic knee osteoarthritis who visited the Department of Orthopedics for a potential knee surgery were evaluated for onychomycosis by PCR-based reverse blot hybridization assay. Of the 520 patients, 308 (59.2%) were diagnosed with onychomycosis. Age (p = 0.004), male sex (p = 0.015), and being barefooted (p = 0.031) were statistically significant risk factors for onychomycosis. Knee disease severity, based on Kellgren-Lawrence grade, was associated with severity of onychomycosis. The impairment of physical function and self-care caused by knee disorders may increase the prevalence of onychomycosis in these patients.

Facile synthesis of economical feasible fly ash-based zeolite-supported nano zerovalent iron and nickel bimetallic composite for the potential removal of heavy metals from industrial effluents.

Heavy metals contamination of water is one of the environmental issue globally. Thus prepared fly ash-based zeolite (FZA)-supported nano zerovalent iron and nickel (nZVI/Ni@FZA) bimetallic composite from low-cost fly ash waste for the potential treatment of anion (Cr(VI) and cation Cu(II)) heavy metals from industrial effluents at pH 3 and 5, respectively in this study. The systematic interaction between FZA and nZVI/Ni and the adsorptive removal mechanism was studied. The mean surface area of the nZVI/Ni@FZA (154.11 m2/g) was much greater than that of the FZA (46.6 m2/g) and nZVI (4.76 m2/g) independently, as determined by BET-N2 measurements. The effect of influence factors on the removal of Cr(VI) and Cu(II) by nZVI/Ni@FZA, such as pH effect, initial concentration effect, time effect, temperature effect, coexisting metals, and ionic strength, and cumulative loading ability, were discussed. The maximum adsorption capacity of nZVI/Ni@FZA was 48.31 mg/g and 147.06 mg/g towards Cr(VI) and Cu(II), respectively. These were higher than those of nZVI@FZA and FZA. It demonstrated that Ni could play an important role in enhancing the reduction ability of nZVI. Furthermore, isothermal and kinetic results revealed that both heavy metal adsorption processes were rate limiting monolayer Langmuir adsorption on homogeneous surfaces. Thermodynamic results suggested that the adsorptive removal of metal ions was endothermic with spontaneity. The applicability of nZVI/Ni@FZA on real industrial wastewater treatment results demonstrate that the concentration of heavy metals were removed under the acceptable standard levels. Further the adsorption capacity of nZVI/Ni@FZA was higher than the nZVI@FZA and FZA. The overall results demonstrated that nZVI/Ni@FZA was a promising, efficient, and economically feasible sorbent for potential wastewater treatment. Moreover this is first report on the preparation nZVI/Ni@FZA bimetallic composite.

Enhancement of Liposomal Plasmid DNA and siRNA Delivery by Itraconazole through Intracellular Cholesterol Accumulation.

PURPOSE: Efficient and safe vehicle that can enhance gene transfer is still needed. Since intracellular cholesterol is known to have an important role in gene delivery and itraconazole alters intracellular cholesterol trafficking, we investigated the effect of itraconazole on pDNA and siRNA delivery. METHODS: The pDNA and Bcl2 siRNA transfection efficiency was measured by luciferase assay and cytotoxicity. Cellular cholesterol was observed using filipin staining, and intracellular uptake was analyzed by flow cytometry. Lipoplex localization was observed by fluorescent labeling of DNA and lysosome after treatment of itraconazole or co-treatment of itraconazole and bafilomycin A1. RESULTS: Itraconazole enhanced the transfection efficiency of pDNA and siRNA compared to that of control through the accumulation of cholesterol. Bafilomycin A1 diminished the effect of itraconazole on gene delivery and the increment of cholesterol. Itraconazole did not increase the cellular uptake of lipoplex, but increased free pDNA during the endosome-lysosome pathway was observed during the endosome-lysosome pathway. Treating cells with both imipramine and itraconazole caused an additive effect in pDNA and siRNA delivery. CONCLUSIONS: Itraconazole enhanced gene delivery of pDNA and siRNA, and it can be used to potentiate nucleic acid therapeutics.

Clinical scoring system for the prediction of survival of patients with advanced gastric cancer.

OBJECTIVE: In this study, we established a risk scoring system using easily obtained clinical characteristics at the time of initiating palliative chemotherapy to predict accurate overall survival of patients with advanced gastric cancer after first-line treatment with fluoropyrimidine-platinum combination chemotherapy. METHODS: A total of 1733 patients treated at the Samsung Medical Center, Korea were included in the study, and clinicopathological and laboratory data were retrospectively analysed. The dataset was split into a training set (n=1156, 67%) and a validation set (n=577, 33%). Top-ranked variables were identified using the random forest survival algorithm and integrated into a Cox regression model, thereby constructing the scoring system for predicting the overall survival of patients with advanced gastric cancer. RESULTS: The following five variables were finally included in the scoring system: serum neutrophil-lymphocyte ratio, alkaline phosphatase level, albumin level, performance status and histologic differentiation. The scoring system determined four distinct risk groups in the validation dataset with median overall survival of 17.1 months (95% CI=14.9 to 20.5 months), 12.9 months (95% CI=11.4 to 14.6 months), 8.1 months (95% CI=5.3 to 12.3 months) and 3.9 months (95% CI=1.5 to 8.2 months), respectively. The area under the curve to estimate the discrimination performance of the scoring system was 66.1 considering 1 year overall survival. CONCLUSIONS: We developed a simple and clinically useful predictive scoring model in a homogeneous population with advanced gastric cancer treated with fluoropyrimidine-containing and platinum-containing chemotherapy. However, additional independent validation will be required before the scoring model can be used commonly.

Necrobiotic Xanthogranuloma Coexists with Diffuse Normolipidemic Plane Xanthoma and Multiple Myeloma.

Necrobiotic xanthogranuloma (NXG), is a rare multisystem disease that manifests as cutaneous inflammatory lesions, and is commonly associated with lymphoproliferative disease. Diffuse normolipemic plane xanthoma (NX), is also a rare, acquired disease that is often associated with systemic diseases such as lymphoproliferative disease. Both of these diseases have been reported to be associated with monoclonal gammopathy (MG). However, there are few cases in which these diseases co-exist. A 78-year-old female, who had a known case of NX on the neck and axillary area, presented with an asymptomatic erythematous plaque on her left supraclavicular area. Histopathological examination showed lymphoid aggregates, necrobiotic areas, and granulomatous inflammation in the dermis. Numerous foreignbody and Touton type giant cells were noticed. Serum protein immunoelectrophoresis showed an IgG kappa type MG. Lipid profile of the patient was normal. Bone marrow examination showed plasma cell myeloma. Based on these histologic and laboratory results, we diagnosed this lesion as NXG coexisting with NX and multiple myeloma. She was started on treatment with bortezomib and melphalan for multiple myeloma, and high-dose systemic corticosteroid and triamcinolone intralesional injection for the skin lesion. After 3 months of treatment, the NXG skin lesion and MG improved.

Combination of Doxorubicin with Gemcitabine-Incorporated G-Quadruplex Aptamer Showed Synergistic and Selective Anticancer Effect in Breast Cancer Cells.

Doxorubicin (DOX) is one of the most effective anticancer agents used for the treatment of multiple cancers; however, its use is limited by its short half-life and adverse drug reactions, especially cardiotoxicity. In this study, we found that the conjugate of DOX with APTA12 (Gemcitabine incorporated G-quadruplex aptamer) was significantly more cancer selective and cytotoxic than DOX. The conjugate had an affinity for nucleolin, with higher uptake and retention into the cancer cells than those of DOX. Further, it was localized to the nucleus, which is the target site of DOX. Owing to its mechanism of action, DOX has the ability to intercalate into the nucleotides thus making it a suitable drug to form a conjugate with cancer selective aptamers such as APTA12. The conjugation can lead to selectively accumulate in the cancer cells thus decreasing its potential nonspecific as well as cardiotoxic side effects. The aim of this study was to prepare a conjugate of DOX with APTA12 and assess the chemotherapeutic properties of the conjugate specific to cancer cells. The DOX-APTA12 conjugate was prepared by incubation and its cytotoxicity in MCF-10A (non-cancerous mammary cells) and MDA-MB-231 (breast cancer cells) was assessed. The results indicate that DOX-APTA12 conjugate is a potential option for chemotherapy especially for nucleolin expressing breast cancer with reduced doxorubicin associated side effects.

Variability in Doctors' Usage Paths of Mobile Electronic Health Records Across Specialties: Comprehensive Analysis of Log Data.

BACKGROUND: With the emergence of mobile devices, mobile electronic health record (mEHR) systems have been utilized by health care professionals (HCPs), including doctors, nurses, and other practitioners, to improve efficiency at the point of care. Although several studies on mEHR systems were conducted, including those analyzing their effects and HCPs' usage frequency, only a few considered the specific workflows of doctors based on their specialties in which the work process differs greatly. OBJECTIVE: This study aimed to investigate the differences in mEHR usage paths across clinical specialties. METHODS: We collected the log data of 974 doctors who worked from August 2016 to August 2017 and used the mEHR system at the Samsung Medical Center, one of the biggest hospitals in South Korea. The doctors were classified into 3 groups based on their specialty: the physician, the surgeon, and other hospital-based physician (OHBP) groups. We used various descriptive and visualization methods to understand and compare doctors' usage paths of mEHRs. First, the average numbers of log-ins per day and features used per log-in were examined over different specialties and positions. Second, the number of features used by each doctor was visualized via a heat map to provide an overview of mEHR usage across feature types and doctors' specialties. Third, we conducted a path analysis via a Sankey diagram to describe main usage paths and association rule mining to find frequent paths in mEHR usage. RESULTS: The physician group logged on most frequently, whereas the OHBP group logged on least frequently. In fact, the number of log-ins per day of residents in the physician group was 4.4 times higher than that of staff members in the other groups. The heat map visualization showed a visible difference among specialty groups. The physician group used more consultation-related features, whereas the surgeon group used more surgery-related features. Generally, 50% of the doctors spent about 15 seconds at a time when using mEHRs. In the Sankey diagram, the physician group showed diverse usage patterns with higher complexity compared with the other 2 groups; in particular, their paths contained more loops, which reflected repetitive checks on multiple patients. The most frequent path included inpatient summary, which means that most users stopped at the point of summary and did not proceed to view more details. CONCLUSIONS: The usage paths of mEHRs showed considerable differences among the specialty groups. Such differences can be accommodated into an mEHR design to enhance the efficiency of care.

The First Reported Case of Fibrous Hamartoma of Infancy with Hyperhidrosis and Hypertrichosis in Korea.

Fibrous hamartoma of infancy (FHI) is a rare entity with a benign nature. The typical clinical features are a single, slowly growing, painless mass on the trunk that appears within the first 2 years of life. We report a 13-month-old boy who presented with a plaque on the lower back since 4 months of age. The plaque had gradually become larger and firm, and hyperhidrosis and hypertrichosis were noticed. No visible connection between the spinal cord and the lesion was found in radiologic studies, indicating a disease other than spinal dysraphism. Histopathological findings showed well-defined fibrous trabeculae, mature adipose tissue, and primitive mesenchymal cells, all consistent with FHI. This is the first case of FHI presenting with hyperhidrosis and hypertrichosis reported in Korea.

Synthesis of cholesteryl doxorubicin and its anti-cancer activity.

Doxorubicin (dox) has been used as anti-cancer agent, but there are disadvantages such as rapid excretion, short retention time and cardiotoxicity. For giving lipophilic properties to dox, it was modified with cholesterol derivatives that were validated as a component of liposomal gene delivery. This article describes the synthesis of dox derivatives (lipo-dox A-D), their cytotoxicity and cellular uptake. In A549, HeLa, MCF7 and MDA MB 231 cell lines, lipo-dox A and lipo-dox B substituted at alcohol group showed similar anti-cancer effect as dox, but lipo-dox C and lipo-dox D substituted at amino group did not. As a result, the amino group of dox seems an important site for its cancer cell inhibition. Lipophilic property of lipo-dox A and lipo-dox B induced more accumulation in cells compared to parent drug. Therefore, the newly synthesized lipo-dox A and lipo-dox B would be a good candidate for anti-cancer agent.

Immunohistochemical expression of cytokeratin 15, cytokeratin 19, follistatin, and Bmi-1 in basal cell carcinoma.

BACKGROUNDS: Basal cell carcinoma (BCC) is the most common cutaneous neoplasm. Despite numerous previous studies, the origin of BCC is still controversial. OBJECTIVE: The aim of this study is to evaluate whether BCC arises from the hair follicle rather than the epidermal basal cell. METHODS: The labeling index and differences of expression of the immunohistochemical (IHC) stainings of CK15, CK19, follistatin, and Bmi-1 were evaluated in 20 cases of BCC, 13 cases of trichoblastoma, 21 cases of actinic keratosis (AK), and 19 cases of squamous cell carcinoma (SCC) which were biopsied in the department of dermatology of the Yeungnam University Medical Center. RESULTS: For CK15, labeling indexes of BCC (83.0%) and trichoblastoma (84.4%) were significantly higher than those of AK (15.9%) and SCC (15.8%). For CK19, labeling indexes of BCC (8.1%), trichoblastoma (6.6%), AK (3.5%), and SCC (14.8%) revealed no difference. For follistatin, labeling indexes of BCC (51.1%) and trichoblastoma (70.1%) were significantly higher than those of AK (0.9%) and SCC (8.5%). For Bmi-1, labeling indexes of BCC (74.4%) and trichoblastoma (84.7%) were significantly higher than those of AK (24.7%) and SCC (18.6%). CONCLUSION: In this study, strong positivity of CK15, follistatin, and Bmi-1 was more frequently observed in BCC and trichoblastoma than in SCC and AK. These findings suggest that BCC and trichoblastoma share the same differentiation toward the hair follicle. Furthermore, CK15, follistatin, and Bmi-1 can be useful as markers to differentiate BCC from SCC.

Epidemiology of deep cutaneous fungal infections in Korea (2006-2010).

Deep cutaneous fungal infections (DCFI) occur worldwide and their prevalence is influenced by personal factors of the affected patients and the geographic and cultural features. Surveillance studies of DCFI with respect to the various clinical backgrounds of affected patients can ultimately help to improve their outcome. Expanding on our previous study, we performed a retrospective analysis of patients with DCFI who were treated in a group of university teaching hospitals in Korea to determine the trends within a 5-year period. A retrospective medical record review of patients with DCFI treated between 2006 and 2010 at 16 university teaching hospitals located throughout Korea was performed. Among the 51 cases of DCFI (median patient age, 47.0 years), opportunistic infections in immunocompromised hosts accounted for half. Patients in this group included 11 who were transplant recipients and 12 with malignancies. Overall, Candida (13/51) was the most common causative organism, followed by Sporothrix (12) and Aspergillus (6). Papuloplaques and nodular lesions were the typical presentation, with maculopatches and ulcers also occurring in considerable numbers. Ten patients had systemic involvement. Eight immunocompromised patients did not recover from the disease despite systemic antifungal treatment. Our results highlight the equal involvement of opportunistic and primary pathogens in DCFI, as determined in cases from a 5-year period. Especially in immunocompromised hosts with non-specific skin findings, clinical suspicion is important because failure to diagnose a DCFI causes significant morbidity and possibly even death.

DOTAP/DOPE ratio and cell type determine transfection efficiency with DOTAP-liposomes.

The effects of lipid compositions on their physicochemical properties and transfection efficiencies were investigated. Four liposome formulations with different 1,2-dioleoyl-3-trimethylammoniumpropane (DOTAP) to dioleoylphosphatidylethanolamine (DOPE) weight ratios were investigated, that is, weight ratios 1:0 (T1P0), 3:1 (T3P1), 1:1 (T1P1), and 1:3 (T1P3). Mean sizes of liposomes were influenced by their lipid composition and the preparation concentration at the time of sonication. Zeta potentials of liposomes were inversely correlated with their liposome sizes. However, neither liposome sizes nor zeta potentials were correlated with transfection efficiency. The optimum composition of liposomes was cell-line dependent (T1P0 and T3P1 for Huh7 and AGS, T3P1 and T1P1 for COS7, and T1P1 and T1P3 for A549). The shape of lipoplexes was changed from lamellar to inverted hexagonal structure according to the increased ratio of DOPE, but there was no definite advantage of specific structure in transfection efficiency throughout all used cell lines. However, cellular internalization was consistently faster in T1P0, T3P1, T1P1 compared to T1P3 in all cell lines, suggesting the importance of endosomal escape. Our findings show that the transfection efficiency of DOTAP liposomes is mainly influenced by lipid composition and cell type, and not by size or zeta potential.