Characteristics of Attention-Deficit/Hyperactivity Disorder Subtypes in Children Classified Using Quantitative Electroencephalography.

Purpose: This study used quantitative electroencephalography (QEEG) to investigate the characteristics of attention-deficit/hyperactivity disorder (ADHD) subtypes in children. Patients and Methods: There were 69 subjects (42 with ADHD and 27 neurotypical (NT)) in this study. A semi-structured interview was conducted with each participant for psychiatric diagnostic evaluation. We measured the absolute and relative power in 19 channels and analyzed QEEG using the following frequency ranges: delta (1-4 Hz), theta (4-8 Hz), alpha 1 (8-10 Hz), alpha 2 (10-12 Hz), beta 1 (12-15 Hz), beta 2 (15-20 Hz), beta 3 (20-30 Hz), and gamma (30-45 Hz). Group analyses and EEG noise preprocessing were conducted using iSyncBrain, a cloud-based, artificial intelligence EEG analysis platform. Analysis of covariance adjusted for IQ, age, and sex was used. Results: QEEG analysis revealed three ADHD subtypes, characterized by (A) elevated relative fast alpha and beta power, (B) elevated absolute slow frequency (delta and theta power), or (C) elevated absolute and relative beta power. A significant difference was found in the Korean ADHD Rating Scale (K-ARS) among the four groups (df=3, F=8.004, p<0.001); group C had the highest score (25.31±11.16), followed by group A (21.67±13.18). The score of group B (12.64±7.84) was similar to that of the NT group (11.07±6.12) and did not reach the cut-off point of the K-ARS. In the Wender-Utah Rating Scale (WURS), group B score (55.82±23.17) was significantly higher than the NT group score (42.81±13.26). Conclusion: These results indicate that children with ADHD do not constitute a neurophysiologically homogenous group. Children with QEEG subtype B (elevated slow frequency) may be difficult to distinguish from normal children using the K-ARS, which is the most common screening tool for ADHD. Moreover, parents of children with this subtype may be less sensitive to observing ADHD symptoms.

The Cytoskeletal Transport Protein, Secretagogin, Is Essential for Diurnal Glucagon-like Peptide-1 Secretion in Mice.

The intestinal L-cell incretin, glucagon-like peptide-1 (GLP-1), exhibits a circadian pattern of secretion, thereby entraining diurnal insulin release. Secretagogin (Scgn), an actin-binding regulatory protein, is essential for the temporal peak of GLP-1 secretion in vitro. To interrogate the role of Scgn in diurnal GLP-1 secretion in vivo, peak and trough GLP-1 release were evaluated in knockout mice (Scgn-/-, Gcg-CreERT2/+; Scgnfl/fl and Vil-CreERT2/+; Scgnfl/fl), and RNA sequencing (RNA-Seq) was conducted in Scgn knockdown L-cells. All 3 knockout models demonstrated loss of the diurnal rhythm of GLP-1 secretion in response to oral glucose. Gcg-CreERT2/+; Scgnfl/fl mice also lost the normal pattern in glucagon secretion, while Scgn-/- and Vil-CreERT2/+; Scgnfl/fl animals demonstrated impaired diurnal secretion of the related incretin, glucose-dependent insulinotrophic polypeptide. RNA-Seq of mGLUTag L-cells showed decreased pathways regulating vesicle transport, transport and binding, and protein-protein interaction at synapse, as well as pathways related to proteasome-mediated degradation including chaperone-mediated protein complex assembly following Scgn knockdown. Scgn is therefore essential for diurnal L-cell GLP-1 secretion in vivo, likely mediated through effects on secretory granule dynamics.

Disrupted and Elevated Circadian Secretion of Glucagon-Like Peptide-1 in a Murine Model of Type 2 Diabetes.

Metabolism and circadian rhythms are intimately linked, with circadian glucagon-like peptide-1 (GLP-1) secretion by the intestinal L-cell entraining rhythmic insulin release. GLP-1 secretion has been explored in the context of obesogenic diets, but never in a rodent model of type 2 diabetes (T2D). There is also considerable disagreement regarding GLP-1 levels in human T2D. Furthermore, recent evidence has demonstrated decreased expression of the ß-cell exocytotic protein secretagogin (SCGN) in T2D. To extend these findings to the L-cell, we administered oral glucose tolerance tests at 6 time points in 4-hour intervals to the high-fat diet/streptozotocin (HFD-STZ) mouse model of T2D. This revealed a 10-fold increase in peak GLP-1 secretion with a phase shift of the peak from the normal feeding period into the fasting-phase. This was accompanied by impairments in the rhythms of glucose, glucagon, mucosal clock genes (Arntl and Cry2), and Scgn. Immunostaining revealed that L-cell GLP-1 intensity was increased in the HFD-STZ model, as was the proportion of L-cells that expressed SCGN; however, this was not found in L-cells from humans with T2D, which exhibited decreased GLP-1 staining but maintained their SCGN expression. Gcg expression in isolated L-cells was increased along with pathways relating to GLP-1 secretion and electron transport chain activity in the HFD-STZ condition. Further investigation into the mechanisms responsible for this increase in GLP-1 secretion may give insights into therapies directed toward upregulating endogenous GLP-1 secretion.

The Relationship between the Drug Delivery Properties of a Formulation of Teriparatide Microneedles and the Pharmacokinetic Evaluation of Teriparatide Administration in Rats.

PURPOSE: Teriparatide is an effective drug for the treatment of osteoporosis. This study examines the relationship between the drug delivery properties of the solid formulation with teriparatide and the pharmacokinetic properties of teriparatide in vivo. METHODS: Teriparatide microneedles with different dissolution rates were prepared using sucrose and carboxymethylcellulose (CMC). There were three aspects of this study: (1) The dissolution rate of teriparatide from both formulations (sucrose and CMC) was measured in vitro. (2) After administration into porcine skin ex vivo, the diffusion rate of FITC-dextran was observed using a confocal microscope. (3) Pharmacokinetic studies were performed in rats and pharmacokinetic data compared with the release rate and the diffusion pattern. RESULTS: In the in vitro dissolution experiment, 80% of teriparatide was released within 30 min from the CMC MNs, whereas 80% of teriparatide was released within 10 min from the sucrose MNs. After 30 min, the fluorescence intensity on the surface of the MNs was 40% of the initial intensity for sucrose MNs and 90% for CMC MNs. In the pharmacokinetic study, the Cmax values of the CMC and sucrose MNs were 868 pg/mL and 6809 pg/mL, respectively, and the AUClast values were 6771 pg*hr/mL for the CMC MNs and 17,171 pg*hr/mL for the sucrose MNs. CONCLUSIONS: When teriparatide is delivered into the skin using microneedles, the release rate from the solid formulation determines the drug's pharmacokinetic properties. The diffusion pattern of fluorescence into the skin can be used to anticipate the pharmacokinetic properties of the drug.

Wireless graphene-based thermal patch for obtaining temperature distribution and performing thermography.

Thermal imaging provides information regarding the general condition of the human body and facilitates the diagnosis of various diseases. Heat therapy or thermotherapy can help in the treatment of injuries to the skin tissue. Here, we report a wearable thermal patch with dual functions of continuous skin temperature sensing and thermotherapy for effective self-care treatment. This system consists of a graphene-based capacitive sensor, a graphene thermal pad, and a flexible readout board with a wireless communication module. The wearable sensor continuously monitors the temperature variation over a large area of the skin (3 × 3cm2) with high resolution and sensitivity and performs thermotherapy via the graphene-based heater mounted at the bottom of the device. Animal studies prove that the proposed system can be used to diagnose various diseases. This technology could be useful in the development of convenient and wearable health care devices.

Complementary and antagonistic effects of combined glucagon-like peptide-2 and glucagon-like peptide-1 receptor agonist administration on parameters relevant to short bowel syndrome.

BACKGROUND: Short bowel syndrome (SBS) is characterized by malabsorption requiring parenteral nutrition. The intestinotrophic glucagon-like peptide (GLP)-2 receptor agonist, h[Gly2]GLP2, is used to treat patients with SBS. Evidence suggests that GLP-1 receptor agonists such as exendin-4 (Ex4) may be beneficial in SBS given their ability to increase intestinal growth and delay gastric emptying (GE). METHODS: Intestinal growth, body weight (BW), food intake (FI), GE, gastrointestinal (GI) transit, intestinal permeability, and glucose tolerance were investigated in male and female C57/BL6 mice following vehicle, h[Gly2]GLP2, or Ex4 treatment, alone or in combination at "low," "medium," and "high" doses (0.1, 0.5, 1.0 and 0.01, 0.05, 0.1 µg/g, respectively). RESULTS: Only the h[Gly2]GLP2 low/Ex4 high-dose combination additively increased small intestinal (SI) weight compared with vehicle and both monoagonists (P < 0.01-0.001), via increased villus height (P < 0.01) and SI length (P < 0.05). This combination had no effects on BW; FI; and fat, liver, spleen, heart, and kidney weights but reduced GI transit (P < 0.001) versus low-dose h[Gly2]GLP2 monotreatment and abrogated the inhibitory effects of high-dose Ex4 on GE (P < 0.01) and of low-dose h[Gly2]GLP2 on intestinal permeability (P < 0.05). Ex4-induced improvements in glucose homeostasis were maintained upon combination with h[Gly2]GLP2 (P < 0.001). CONCLUSIONS: These findings suggest that combining specific doses of GLP-2- and GLP-1 receptor agonists additively improves SI growth and GI transit without detrimental effects on BW, FI, GE, and glucose homeostasis, and may be useful for the treatment of patients with SBS.

Proteasomal activator 28 gamma stabilizes hepatitis B virus X protein by competitively inhibiting the Siah-1-mediated proteasomal degradation.

Proteasomal activator 28 gamma (PA28γ) upregulates the levels of HBx, a regulatory protein of hepatitis B virus (HBV) to stimulate HBV replication; however, the detailed mechanism remains unknown. Here, we found that PA28γ impaired the ability of seven in absentia homolog 1 (Siah-1) as an E3 ubiquitin ligase of HBx. PA28γ competitively inhibited the binding of Siah-1 to HBx in human hepatoma cells. Accordingly, PA28γ increased the stability of HBx and decreased HBx ubiquitination, abolishing the potential of Siah-1 to downregulate HBx levels. PA28γ also executed its role as an antagonist of Siah-1 during HBV replication, as demonstrated by an in vitro HBV replication system. The present study may provide insights into the mechanisms underlying the regulation of HBV replication.

Impairment of synaptic plasticity and novel object recognition in the hypergravity-exposed rats.

The gravity is necessary for living organisms to operate various biological events including hippocampus-related functions of learning and memory. Until now, it remains inconclusive how altered gravity is associated with hippocampal functions. It is mainly due to the difficulties in generating an animal model experiencing altered gravity. Here, we demonstrate the effects of hypergravity on hippocampus-related functions using an animal behavior and electrophysiology with our hypergravity animal model. The hypergravity (4G, 4 weeks) group showed impaired synaptic efficacy and long-term potentiation in CA1 neurons of the hippocampus along with the poor performance of a novel object recognition task. Our studies suggest that altered gravity affects hippocampus-related cognitive functions, presumably through structural and functional adaptation to various conditions of gravity shift.

Safe Coated Microneedles with Reduced Puncture Occurrence after Administration.

The goal of this study is the preparation of safer coated microneedles so that tips remaining after the initial use are less likely to be reinserted on a second use. Twelve groups of uncoated microneedles (u-MNs) were prepared from the combination of three different aspect ratios (height to base width) and four kinds of polymer (polyethylene (PE), polypropylene (PP), nylon and polylactic acid (PLA)). After coating the u-MNs with polyvinyl alcohol formulation to make coated MNs (c-MNs), the force displacement of the u-MNs and the c-MNs was measured. The aspect ratio was reduced from 2.2, 2.5 and 3.0 with u-MNs to 1.3, 1.4 and 1.6 with c-MNs, respectively, after the coating formulation was applied to the MNs. All PLA MNs had a puncture performance of more than 95%. However, the puncture performance of u-MNs made of PE and of PP with a 3.0 aspect ratio was only 8% and 53%, respectively, whereas the rates of c-MNs made of PE and of PP were 82% and 95%, respectively. In animal experiments with PP MNs with a 3.0 aspect ratio, the 59% rate of puncture performance with u-MNs increased to above 96% with c-MNs and fell to 13% for r-MNs. Safe c-MNs can overcome the disadvantages of standard c-MNs by reducing the probable contamination of remaining tips after use. Safe c-MNs have advantages over standard c-MNs in terms of humidity resistance, reasonable cost, sterilization process and short processing time through the separate process of u-MN preparation and simple dip-coating.

Diffractive multifocal intraocular lens implantation in patients with monofocal intraocular lens in the contralateral eye.

AIM: To evaluate clinical outcomes of unilateral implantation of a diffractive multifocal intraocular lens (IOL) in patients with contralateral monofocal IOL. METHODS: Twenty-two patients who already had implantation of a monofocal IOL in unilateral eye underwent implantation of a diffractive multifocal IOL in contralateral eye were enrolled. After 1, 6, and 12mo, uncorrected and distant corrected distant visual acuity (UCDVA and DCDVA), uncorrected and distant corrected intermediate-visual acuity (UCIVA and DCIVA), uncorrected and distant corrected near visual acuity (UCNVA and DCNVA), and contrast sensitivity were obtained. Halo/glare symptoms, spectacle dependence, and patient satisfaction were also evaluated. RESULTS: The mean age was 67.86±7.25y and the average interval between two IOL implantations was 645.82±878.44d. At 1mo, binocular UCDVA was lower than 0.20 logMAR in 76% of patients (mean 0.12±0.13 logMAR), which increased to 90% by 6 and 12mo. The binocular UCDVA was significantly better than the monocular results (P<0.05) at 1, 6, and 12mo. Additionally, UCNVA was lower than 0.40 logMAR in 82% of patients, increasing to 90% by 6 and 12mo. Mean UCNVA in the multifocal IOL implanted eye was statistically significantly better than that in the monofocal IOL implanted eye (P<0.05) at 1, 6, and 12mo. About 5% of patients at 1 and 6mo, reported "severe glare or halo". Patient satisfaction rates were 95% and 91% at 6 and 12mo, respectively. CONCLUSION: Unilateral implantation of multifocal IOL in patients with a contralateral, monofocal IOL implantation results in high patient satisfaction rate, with low severe glare or halo rate during follow-up. It can represent a good option for patients who have previously had a monofocal IOL implantation regardless of two year interval duration between two IOL implantations.

Preclinical study of influenza bivalent vaccine delivered with a two compartmental microneedle array.

Multiple vaccines can be mixed into a single combination to be a single product. However, combination vaccines have problems of complexity. In this study, microneedles were utilized in a compartmental microneedle array (CMA) to deliver two influenza vaccine strains without mixing. In this study, the CMA had two compartments, and two rectangular structures were attached to each end of the array to enable integration of the compartments with the coating equipment. The coating solution, which contained influenza vaccines for B/Yamagata (B-Y) and B/Victoria (B-V), was filled into the two reservoirs of the container. The CMA was aligned with the container for dipping the first compartment of the array into the first reservoir and the second compartment into the second reservoir. The CMA containing B-Y and B-V separately was administered to mice, and weight change and survival were compared with other groups of mice administered (a) combination vaccines with microneedles, (b) two monovalent vaccines with microneedles, (c) intramuscularly with a combination vaccine, and (d) intramuscularly with two monovalent vaccines. Plaque reduction neutralization tests were also performed to compare the CMA group with the other groups. The CMA showed a relative standard error of less than 7% between samples in dose uniformity. It also showed comparable antibody-forming efficacy compared to other groups, especially by B/Yamagata virus challenge. The CMA mice group showed better survival and weight change than mice that received intramuscular (IM) injection of the combination vaccine. In the neutralizing antibody experiment, all microneedle groups showed a higher neutralizing antibody than the IM groups. Vaccines were administered without mixing by a single administration using a CMA, and the CMA showed comparable efficacy with IM administration of the combination vaccine. Multivalent vaccines can be delivered without mixing as a single product by using a CMA.

Ultrasound-assisted gatifloxacin delivery in mouse cornea, in vivo.

Gatifloxacin is a 4th generation fluoroquinolone antibiotic used in the clinic to treat ocular infection. One limitation of gatifloxacin is its relatively poor corneal penetration, and the increase of its trans-corneal delivery would be beneficial to reduce the amount or frequency of daily dose. In this study, ultrasound treatment was applied to enhance the trans-corneal delivery of gatifloxacin without damage. Experiments were conducted on mouse eyes in ex vivo and in vivo conditions. Ultrasound waves with 1 MHz in frequency, 1.3 W/cm2 in intensity were applied onto the mouse cornea for 5 minutes, and then gatifloxacin ophthalmic solution was instilled and left there for 10 minutes. 3D gatifloxacin distribution in the cornea was measured by two-photon microscopy (TPM) imaging based on its intrinsic fluorescence. Longitudinal TPM imaging of ultrasound treated mouse corneas showed the increase of initial gatifloxacin intensities on the corneal surface compared to untreated mouse corneas by 67%, and then the increased gatifloxacin delivery into the cornea from the surface at later time. The delivered gatifloxacin in the corneal epithelium stayed longer in the ultrasound treated corneas than in the untreated corneas. The enhanced trans-corneal delivery and extended stay of gatifloxacin in the mouse cornea by ultrasound treatment could be beneficial for therapeutic effects. This study demonstrated the detail process of enhanced trans-corneal gatifloxacin delivery by ultrasound treatment.

HBx natural variants containing Ser-101 instead of Pro-101 evade ubiquitin-dependent proteasomal degradation by activating proteasomal activator 28 gamma expression.

Proteasomal activator 28 gamma (PA28γ) is frequently overexpressed in hepatocellular carcinoma; however, its underlying mechanism and role in hepatitis B virus (HBV) replication are largely unknown. Here, we found that HBV X protein (HBx) natural variants containing Ser-101 instead of Pro-101 increase reactive oxygen species levels in the mitochondria and activate the ataxia telangiectasia mutated/checkpoint kinase 2 pathway in the nucleus, resulting in the phosphorylation of p53 at Ser-15 and Ser-20 and the subsequent upregulation of its protein levels. Therefore, HBx variants containing Ser-101 induced p53-dependent activation of PA28γ expression in human hepatoma cells. The elevated PA28γ levels upregulated HBx levels through the inhibition of seven in absentia homologue 1-dependent proteasomal degradation. The self-amplifying ability of HBx variants containing Ser-101 via a positive feedback loop involving p53 and PA28γ was accurately reproduced in both a 1.2-mer HBV replicon and in vitro HBV infection systems, which also provided evidence for the stimulation of HBV replication by these HBx variants. In conclusion, the ability of HBx to upregulate PA28γ levels via p53 activation, in a Ser-101-dependent pathway, is critical for the stimulation of HBV replication.

Development of Botulinum Toxin A-Coated Microneedles for Treating Palmar Hyperhidrosis.

Hyperhidrosis is a disorder that is characterized by the production of excess amounts of sweat. The botulinum neurotoxin A (BoNT/A) has been used to treat hyperhidrosis through multiple intradermal injections at the site of the condition. However, because of BoNT/A toxicity, it is important to precisely deliver the proper dose of the toxin to the target site. In addition, the use of a conventional hypodermic needle for multiple injections in the palm makes the approach undesirable and painful. Here, we designed a BoNT/A-coated microneedle (BoNT-MN) array and tested its efficacy as a substitute pain-free method to treat hyperhidrosis. BoNT-MNs were prepared by coating polylactic acid microneedles with a BoNT/A formulation and were found to successfully penetrate into a thick skin in vitro. The coating formulations were then tested for their stability at 4, 25, and 37 °C for 24 h. BoNT-MNs were found to be much more stable than BoNT/A in a liquid state. Additionally, we carried out in vivo experiments by treating the right paws of mice with BoNT-MNs and found that the treatment induced a significant reduction in the sweating response in the mouse foot pad. Thus, BoNT/A treatment using microneedles is beneficial and may be used as a more efficient and less painful approach to treat hyperhidrosis.

Preparation of H1N1 microneedles by a low-temperature process without a stabilizer.

During the manufacture of H1N1 microneedles, a stabilizer is usually added to maintain the antigenicity of the vaccine. However, finding a suitable stabilizer is difficult, and the addition of a stabilizer can limit the antigen dose and the addition of an adjuvant because of the limited volume of the microneedles. In this study, the authors evaluated whether H1N1 microneedles could be fabricated without a stabilizer by keeping the production environment at a low temperature. H1N1 microneedle patches without a stabilizer were prepared in a process that involved maintaining a low temperature of 10 °C. The protective immune response to this method of drug application was investigated by comparing it with traditional intramuscular (IM) immunization and with the use of H1N1 microneedles with a stabilizer. A process-sensitive antigen, H1N1, was stabilized without the use of a stabilizer in a process that maintained a low temperature of 10 °C. The preparation process consisted of coating and drying processes. In animal experiments, mice were immunized using an array of low-temperature H1N1 microneedles without a stabilizer (LT-MN), and they showed strong antibody responses. Compared to three other application methods of traditional IM immunization, low-temperature H1N1 microneedles with a stabilizer (LT-MN-T), and room-temperature H1N1 microneedles with a stabilizer (RT-MN-T), LT-MN produced comparable results in inducing protective immunity. A plaque reduction neutralization test found that LT-MN and LT-MN-T provided greater immunity compared with IM and RT-MN-T. A process in which the temperature is maintained at 10 °C can provide successful vaccination with H1N1 microneedles without the addition of a stabilizer. This process can be applied to various temperature-sensitive biologics.

Thermogenesis-independent metabolic benefits conferred by isocaloric intermittent fasting in ob/ob mice.

Intermittent fasting (IF) is an effective dietary intervention to counteract obesity-associated metabolic abnormalities. Previously, we and others have highlighted white adipose tissue (WAT) browning as the main underlying mechanism of IF-mediated metabolic benefits. However, whether IF retains its efficacy in different models, such as genetically obese/diabetic animals, is unknown. Here, leptin-deficient ob/ob mice were subjected to 16 weeks of isocaloric IF, and comprehensive metabolic phenotyping was conducted to assess the metabolic effects of IF. Unlike our previous study, isocaloric IF-subjected ob/ob animals failed to exhibit reduced body weight gain, lower fat mass, or decreased liver lipid accumulation. Moreover, isocaloric IF did not result in increased thermogenesis nor induce WAT browning in ob/ob mice. These findings indicate that isocaloric IF may not be an effective approach for regulating body weight in ob/ob animals, posing the possible limitations of IF to treat obesity. However, despite the lack of improvement in insulin sensitivity, isocaloric IF-subjected ob/ob animals displayed improved glucose tolerance as well as higher postprandial insulin level, with elevated incretin expression, suggesting that isocaloric IF is effective in improving nutrient-stimulated insulin secretion. Together, this study uncovers the insulinotropic effect of isocaloric IF, independent of adipose thermogenesis, which is potentially complementary for the treatment of type 2 diabetes.

Hepatitis B Virus X Protein Stimulates Virus Replication Via DNA Methylation of the C-1619 in Covalently Closed Circular DNA.

Methylation of HBV cccDNA has been detected in vivo and in vitro; however, the mechanism and its effects on HBV replication remain unclear. HBx derived from a 1.2-mer HBV replicon upregulated protein levels and enzyme activities of DNA methyltransferase 1 (DNMT1), 3a, and 3b, resulting in methylation of the negative regulatory region (NRE) in cccDNA, while none of these effects were observed with an HBx-null mutant. The HBx-positive HBV cccDNA expressed higher levels of HBc and produced about 4-fold higher levels of HBV particles than those from the HBx-null counterpart. For these effects, HBx interrupted the action of NRE binding protein via methylation of the C-1619 within NRE, resulting in activation of the core promoter. Treatment with 5-Aza-2'dC or DNMT1 knock-down drastically impaired the ability of HBx to activate the core promoter and stimulate HBV replication in 1.2-mer HBV replicon and in vitro infection systems, indicating the positive role of HBx-mediated cccDNA methylation in HBV replication.

Hepatitis B virus X protein activates proteasomal activator 28 gamma expression via upregulation of p53 levels to stimulate virus replication.

Proteasomal activator gamma (PA28γ), frequently overexpressed in hepatocellular carcinoma, is believed to play important roles in tumourigenesis. However, the underlying mechanism of PA28γ overexpression and its possible roles in hepatitis B virus (HBV) replication are largely unknown. In the present study, we found that hepatitis B virus X protein (HBx) activates PA28γ expression by upregulating p53 levels in human hepatoma cells. The elevated PA28γ levels in turn repressed seven in absentia homologue 1 expression via downregulation of p53 levels, thereby inhibiting ubiquitin-dependent proteasomal degradation of HBx, which ultimately led to upregulation of HBx levels. The correlation among HBx, p53 and PA28γ was exactly reproduced in a 1.2-mer HBV replicon system, mimicking the natural course of HBV infection. In particular, knockdown of either p53 or PA28γ in HepG2 cells downregulated HBx levels and thereby inhibited HBV replication, whereas overexpression of p53 or PA28γ in Hep3B cells upregulated HBx levels, which stimulated HBV replication, indicating that p53 and PA28γ act as activators of HBV replication. In conclusion, HBx levels are upregulated via a positive feedback loop involving p53 and PA28γ to stimulate HBV propagation.

Local dermal delivery of cyclosporin A, a hydrophobic and high molecular weight drug, using dissolving microneedles.

Dissolving microneedles (MN) containing cyclosporin A (CyA) were prepared for local delivery of CyA into the dermal layer. The efficacy of using MN to deliver CyA, an insoluble and high molecular weight drug, was observed and compared with oral administration of solubilized CyA. Microneedles containing CyA (CyA MN) were prepared using a closed, low-temperature molding process. The mechanical properties of CyA MN and phase separation were studied regarding the content of CyA. CyA MN were inserted into porcine skin for a predetermined time, and the dissolution and delivered amount of CyA were measured in vitro with an optical microscope and high-performance liquid chromatography (HPLC) analysis of the extracted CyA. A pharmacokinetic study of CyA MN was performed in vivo by administering 10% CyA MN, and the pharmacokinetic profile was compared with that of orally administered CyA. Pyramidal CyA MN (600 µm long, 250 µm wide) were prepared. CyA MN penetrated skin successfully with up to 50% CyA content. When 10% CyA MN were pressed into porcine skin for 60 min, 65% of MN length was dissolved and 34 ±â€¯6.5 µg of CyA in MN was delivered into the skin. Under the same conditions with 10% CyA MN administered to rats, CyA MN showed Tmax of 8 h, Cmax of 15.9 ng/ml, and area under curve (AUC) of 686, compared to Tmax of 2 h, Cmax of 18.205 ng/ml, and AUC of 254 for oral administration of solubilized CyA. A therapeutic dose of CyA for treatment of psoriasis was delivered via MN into the skin layer without solubilization of CyA. Due to the hydrophobic properties and high molecular weight of CyA, the safety of CyA delivery was improved using dissolving microneedles because the slow systemic absorption and local treatment enabled CyA to remain in the skin for a longer time. Microneedles are an effective method with high bioavailability for local dermal delivery of insoluble drugs.

Efficacy of a bleomycin microneedle patch for the treatment of warts.

Warts are a common skin disease caused by infection of the human papilloma virus. Most treatments involving physical destruction of the infected cells, such as cryotherapy and electrocautery, are limited by intense pain, failure, or recurrences. Our aim was to compare the therapeutic effects of a newly developed bleomycin microneedle patch with cryotherapy in the treatment of warts. Forty-two patients with more than two wart lesions were included in the study. The two treatment modalities were randomly applied to different warts on each patient. Treatment efficacy was assessed using the Physician's Global Assessment (PGA) and the Patient's Global Assessment (PaGA). Mean PGA and PaGA scores were not significantly different between cryotherapy and bleomycin microneedle patch treatment. It was also determined that the mean size of all the warts treated with either modality shrank about equally at weeks 8 and 16 after initial treatment. Thus, treatment efficacy of the bleomycin microneedle patch was comparable to that of conventional cryotherapy. According to a visual analogue scale of pain, bleomycin microneedle patch treatment was significantly less painful than cryotherapy (p < .0001). In addition, use of the bleomycin microneedle patch was more tolerable for patients who were reluctant to receive the painful treatment. Thus, the bleomycin microneedle patch can be an effective, convenient, and innovative treatment modality for warts.