High-resolution spiral microfluidic channel integrated electrochemical device for isolation and detection of extracellular vesicles without lipoprotein contamination.

Recent studies have indicated significant correlation between the concentration of immune checkpoint markers borne by extracellular vesicles (EVs) and the efficacy of immunotherapy. This study introduces a high-resolution spiral microfluidic channel-integrated electrochemical device (HiMEc), which is designed to isolate and detect EVs carrying the immune checkpoint markers programmed death ligand 1 (PD-L1) and programmed death protein 1 (PD-1), devoid of plasma-abundant lipoprotein contamination. Antigen-antibody reactions were applied to immobilize the lipoproteins on bead surfaces within the plasma, establishing a size differential with EVs. A plasma sample was then introduced into the spiral microfluidic channel, which facilitated the acquisition of nanometer-sized EVs and the elimination of micrometer-sized lipoprotein-bead complexes, along with the isolation and quantification of EVs using HiMEc. PD-L1 and PD-1 expression on EVs was evaluated in 30 plasma samples (10 from healthy donors, 20 from lung cancer patients) using HiMEc and compared to the results obtained from standard tissue-based PD-L1 testing, noting that HiMEc could be utilized to select further potential candidates. The obtained results are expected to contribute positively to the clinical assessment of potential immunotherapy beneficiaries.

Diffusion tensor imaging findings of optic radiation in patients with putaminal hemorrhage.

OBJECTIVES: Little is known about optic radiation (OR) injury in intracerebral hemorrhage (ICH). We attempted to investigate OR injury in patients with ICH by diffusion tensor imaging (DTI). METHODS: Forty-three consecutive patients with putaminal hemorrhage and 40 normal healthy control subjects were recruited. DTI data were acquired at the beginning of rehabilitation (average 34 days after onset). DTI-Studio software was used to reconstruct the OR. Fractional anisotropies (FA) and fiber numbers of the ORs were measured. FA values and fiber numbers of affected ORs were described as abnormal when they were more than 2.5 SD lower than those of normal controls. RESULTS: Thirty (70%) of the 43 patients showed an OR abnormality in the affected hemisphere. In 13 (30%) patients, the affected OR was disrupted or nonreconstructable. On the other hand, of the 20 patients with preserved OR integrity, 14 (33%) had a low FA value and 3 (7%) a low FA and fiber number. The other 13 (30%) of the 43 patients had no abnormal OR findings. CONCLUSION: Seventy percent of patients showed any abnormality of OR in the affected hemisphere on DTI. This result suggests that patients with putaminal hemorrhage are at high risk of OR injury.

Persistent detrusor overactivity in rats after relief of partial urethral obstruction.

Detrusor overactivity (DO) persists after prostatectomy in 20% to 25% of patients with benign disease. Assuming that nonvoiding contractions (NVCs) can be used as a surrogate for DO in humans, the rat model of obstruction/deobstruction may allow us to study the pathophysiology of persistent DO after deobstruction. We investigated bladder function, with a special focus on NVCs, in rats by use of a new, modified method of obstruction and deobstruction and compared these results with those obtained by use of the conventional method. Seventy female Sprague-Dawley rats underwent 1) sham operation (n = 10), 2) obstruction by a modified method (Modif-Obs; n = 12), 3) obstruction/deobstruction by the conventional method (Conv-Obs/Deobs; n = 13), or 4) obstruction/deobstruction by the modified method (Modif-Obs/Deobs; n = 35). The Modif-Obs/Deobs animals were divided into subgroups with (DO+) and without (DO-) NVCs. Two weeks after partial urethral obstruction, the animals were deobstructed, and 1 wk later cystometry was performed with recording of intravesical and intra-abdominal pressures. NVCs were shown in all groups: Modif-Obs (80%), Conv-Obs/Deobs (100%), and Modif-Obs/Deobs (40%). In the Modif-Obs/Deobs group, bladder weight and the muscle-to-collagen ratio were higher in DO+ than in DO- rats. The Modif-Obs/Deobs group showed no mortality compared with 25% mortality in the Conv-Obs/Deobs group. The modified method may be more adequate for studying persistent DO after deobstruction, because it resulted in pressure/volume- and DO-related parameters similar to those found in the clinical situation. The persistence of DO after deobstruction may partly be due to irreversible changes in the bladder caused during the period of obstruction.

Urodynamic findings in an awake chemical cystitis rat model observed by simultaneous registrations of intravesical and intraabdominal pressures.

PURPOSE: The aim of this study was to investigate the effect of urinary bladder inflammation on bladder function in a rat chemical cystitis model. We also histologically confirmed the effects of inflammation in the detrusor on chronically inflamed bladder in rats. MATERIALS AND METHODS: A total of 13 female Sprague-Dawley rats were used in this study. In seven rats, intravesical instillation of HCl induced chemical cystitis, and the other rats with intravesical instillation of saline were used as the sham. After 2 weeks, cystometrograms were obtained with additional intraabdominal pressure measurements in all unanesthetized, unrestrained rats in metabolic cages. The rats were killed just after cystometry. The bladders were removed and examined histologically for mast cells and inflammatory changes. RESULTS: The rats with acute injury by HCl showed no differences in pressure parameters, including basal pressure, threshold pressure, and maximum bladder pressure, compared with the sham rats. They showed significantly increased bladder capacity, micturition volume, residual volume, and micturition interval compared with the sham group. They also showed an increased frequency of detrusor overactivity compared with the sham group. The percent of detrusor overactivity was 56.3% among the total intravesical pressure rises above 2 cmH(2)O. The histological findings of the rats with acute injury by HCl were consistent with chemical cystitis. CONCLUSIONS: Overlapping patterns of lower urinary tract symptoms and pelvic pain are common disease characteristics among interstitial cystitis patients. The situation in an animal model of interstitial cystitis is similar, as observed in this study by the histologic and awake cystometric examinations. However, the interstitial cystitis model showed detrusor overactivity during the filling phase without a decrease in bladder capacity and micturition intervals, which differs from the characteristics of overactive bladder patients.

Selection of a control rat for conscious spontaneous hypertensive rats in studies of detrusor overactivity on the basis of measurement of intra-abdominal pressures.

AIMS: We investigated which animal model is appropriate as a control for the spontaneous hypertensive rat (SHR) in studies of detrusor overactivity (DO). METHODS: Age-matched SHRs (n = 10), Wistar-Kyoto (WKY) rats (n = 9), Wistar-ST (Wistar) rats (n = 10), and Sprague-Dawley (SD) rats (n = 10) were studied. A balloon-fitted catheter was positioned in the abdominal cavity to record intra-abdominal pressure (IAP). Cystometric pressure- and volume-related parameters and DO-related ones during the filling phase were investigated. Of the intravesical pressure (IVP) rises above 2 cmH(2)O, DO was defined as a rise in IVP without a simultaneous change in IAP and was counted during the filling phase. RESULTS: Compared with SHRs, the WKY rats did not show a significant difference in pressure or volume parameters, but Wistar rats and SD rats showed a significant increase in volume parameters, such as bladder capacity, micturition volume, and micturition interval. Typical DO was seen in SHRs and WKY rats during the filling phase, but not in Wistar and SD rats. Furthermore, the frequency and increased amplitude of the DO were similar in WKY rats and SHRs. CONCLUSION: Although WKY rats are an appropriate control for SHRs in studies of hypertension, they may be inappropriate as controls in studies of DO, because they have inherent DO of similar frequency and amplitude as found in SHRs.

Urodynamic Effects of Propiverine on Detrusor Overactivity and Abdominal Straining during Voiding in Awake Rats with Intravesical Prostaglandin E(2) Instillation.

PURPOSE: We investigated the effect of propiverine on cystometric parameters based on intraabdominal pressure (IAP) in awake rats in an overactive bladder (OAB) model induced by intravesical instillation of prostaglandin E(2) (PGE(2)). MATERIALS AND METHODS: Twenty-two female Sprague-Dawley rats were used. Polyethylene catheters were implanted into the bladder to record the intravesical pressure (IVP) and into the femoral artery to administer medication. A balloon-fitted catheter was positioned in the abdominal cavity to record the IAP. Awake cystometries were performed before and after intraarterial administration of propiverine 1 mg/kg (n=6), intravesical administration of 50microM PGE(2) only (n=6), or intravesical PGE(2) plus 1 mg/kg (n=4) or 3 mg/kg (n=6) of intraarterial propiverine. Cystometric pressure and volume parameters and variables related to detrusor overactivity (DO) were investigated. RESULTS: Rats administered intravesical PGE(2) showed increased pressure parameters and decreased volume parameters comparable to the DO model, which was effectively prevented by propiverine (1 or 3 mg/kg). Typical DO shown during the filling phase was decreased by intraarterial propiverine (3 mg/kg) injection. After propiverine (3 mg/kg) injection, IAP was increased at the time of micturition pressure with or without threshold pressure (p<0.05, p<0.01) depending on the dose administered. CONCLUSIONS: Propiverine improved pressure- and volume-related parameters in an OAB model. Furthermore, it also decreased the frequency of DO. However, higher concentrations of propiverine induced straining voiding.

A Case of Skin and Soft Tissue Infection Caused by Mycobacterium abscessus.

Mycobacterium abscessus (M. abscessus) is an acid-fast bacillus that's classified as a pathogenic "rapid growing" nontuberculous mycobacteria. It is an uncommon cause of human disease, but it can cause skin and soft tissue infection after skin injury following inoculation, minor trauma and surgery. The single most important factor for determining the course and prognosis of a M. abscessus infection is the underlying immune status of the host. We report here on a 71-year-old female who presented with multiple painful erythematous cutaneous nodules on her left forearm. She had diabetes mellitus and had taken oral steroid by herself for two years because of her osteoarthritis. Histologically, granulomas and inflammatory cell infiltration were observed and M. abscessus was identified via the mycobacterial culture. We performed curettage and drainage, followed by 6 months of oral clarithromycin and the patient's disease completely healed.

Substantial detrusor overactivity in conscious spontaneously hypertensive rats with hyperactive behaviour.

OBJECTIVE: The spontaneously hypertensive rat (SHR) is a useful model for studying the mechanisms of detrusor overactivity (DO). However, owing to their confounding phenotypic characteristics of hyperactive behaviour, there could be some problems with the interpretation of cystometric data, which is significantly confused by abdominal straining (AS), causing changes in intravesical pressure (IVP). MATERIAL AND METHODS: IVP and intra-abdominal pressure (IAP) were recorded simultaneously to evaluate true DO in conscious healthy male SHRs and Wistar/ST (Wistar) rats. Intravesical pressure rises (IVPRs), defined as increments that exceeded 2 cmH(2)O from baseline, were counted, and those that were caused by AS or DO were determined according to the presence of simultaneous changes in IAP. RESULTS: Compared with Wistar rats, SHRs had a shorter filling phase (p<0.01) and greater frequency of IVPRs (p<0.01). SHRs, but not Wistar rats, showed DO. The substantial DO represented up to 76% of total IVPRs, with the remainder caused by AS. The amplitude (p<0.05) but not the frequency of AS (p>0.05), was higher in SHR than Wistar rats. CONCLUSION: In conscious SHRs, variations in IAP due to hyperactive behaviour may cause misinterpretation of pressure parameters and result in false reports of DO. Thus, simultaneous registration of IAP and IVP is needed for accurate recording of substantial DO in these animals.

Adipose-derived stem cells and their secretory factors as a promising therapy for skin aging.

BACKGROUND: Adipose-derived stem cells (ADSCs) and their secretory factors can stimulate collagen synthesis and migration of fibroblasts during the wound healing process. Conventional treatments for skin aging, such as lasers and topical regimens, induce new collagen synthesis via activation of dermal fibroblasts or growth factors. Considering the results of our previous study, ADSCs can also be used for the treatment of skin aging. OBJECTIVE: The aim was to verify the effectiveness of ADSCs for the treatment of skin aging. METHODS: We analyzed secretory factors of ADSCs and intradermally injected ADSCs (1 x 10(6) cells in 1 mL of Hanks' buffered salt solution) and conditioned media of ADSCs on the back of a micropig. In addition, as a pilot study, intradermal injections of purified autologous processed lipoaspirate (PLA) cells were tried with the photoaged skin of one patient. RESULTS: We demonstrated that ADSCs produce many useful growth factors, increase collagen production in animal study, and reverse skin aging in human trial. CONCLUSION: ADSCs and their secretory factors show promise for application in cosmetic dermatology, especially in the treatment of skin aging.

Medial reorganization of motor function demonstrated by functional MRI and diffusion tensor tractography.

PURPOSE: Peri-lesional reorganization is a motor recovery mechanism after brain injury. The object of this study was to demonstrate the peri-lesional reorganization, using functional MRI (fMRI) and diffusion tensor tractography (DTT). METHODS: Six control subjects and a 53 year-old woman with left primary sensori-motor cortex (SM1) and underlying deep white matter injury were evaluated. The patient presented with complete paralysis of the right hand after the resection of a meningioma on the left SM1. The motor function of the affected hand had recovered slowly the extent of her being able to overcome some resistance for 6 months. At 2 years after the operation, fMRI was performed at 1.5 T in parallel with timed finger flexion-extension movements in all subjects and DTT was performed only in the patient. RESULTS: The contralateral SM1 centered on the precentral knob was activated during hand movements of unaffected (left) side or control subjects. However, the medial area of the injured SM1 was activated during affected (right) hand movements. DTT of the affected (left) hemisphere originated from the medial area of the injured SM1 and descended along the medial side of injured deep white matter. CONCLUSIONS: It seems that the motor function of the affected hand was reorganized into the medial area of the injured SM1 and deep white matter in this patient.