Search

Although inhalation therapy represents a promising drug delivery route for the treatment of respiratory diseases, the real-time evaluation of lung drug deposition remains an area yet to be fully explored. To evaluate the utility of the photo reflection method (PRM) as a real-time non-invasive monitoring of pulmonary drug delivery, the relationship between particle emission signals measured by the PRM and in vitro inhalation performance was evaluated in this study. Symbicort® Turbuhaler® was used as a model dry powder inhaler. In vitro aerodynamic particle deposition was evaluated using a twin-stage liquid impinger (TSLI). Four different inhalation patterns were defined based on the slope of increased flow rate (4.9-9.8 L/s 2 ) and peak flow rate (30 L/min and 60 L/min). The inhalation flow rate and particle emission profile were measured using an inhalation flow meter and a PRM drug release detector, respectively. The inhalation performance was characterized by output efficiency (OE, %) and stage 2 deposition of TSLI (an index of the deagglomerating efficiency, St2, %). The OE × St2 is defined as the amount delivered to the lungs. The particle emissions generated by four different inhalation patterns were completed within 0.4 s after the start of inhalation, and were observed as a sharper and larger peak under conditions of a higher flow increase rate. These were significantly correlated between the OE or OE × St2 and the photo reflection signal (p < 0.001). The particle emission signal by PRM could be a useful non-invasive real-time monitoring tool for dry powder inhalers., (© 2024. The Author(s).)

Chemotherapy-induced oral mucositis (COM) is a common adverse effect of cancer chemotherapy. Several clinical studies reported that repetitive use of mouthwashes containing 2.5-6.25% Hangeshashinto (HST), a Kampo formula, relieves COM, but the effect is insufficient. To solve this problem, we produced an oral ointment of 12% HST extract (considered quantitatively equivalent to 20% commercially available HST), which will increase the local concentrations of its active ingredients and prolong the contact time with COM. In this study, we evaluated the pharmaceutical properties (spreadability and stability) of HST oral ointment. In addition, its safety (oral mucosal irritation) and therapeutic effects on 5-fluorouracil-induced oral mucositis were evaluated in male Syrian hamsters. The HST ointment showed good spreadability and stability for more than 8 weeks at 4 °C. In the oral mucosal irritation test, topical application of HST ointment (0.2 g) three times per day for 14 days had no adverse effect on the oral mucosa of hamsters. In hamsters treated with 5-fluorouracil (60 mg/kg) twice, COM was induced by a submucosal injection of 5% acetic acid into the cheek pouch. When HST ointment (50 µg) was topically applied to the mucositis area once per day for 12 days, the area and macroscopic score of mucositis were significantly decreased, and the depth of the wound tended to be reduced compared with the lactose ointment-treated control animals. These findings suggest that HST oral ointment shows good properties in spreadability, stability, and safety, and elicits a therapeutic effect in an animal model of COM., (© 2022. The Author(s) under exclusive licence to The Japanese Society of Pharmacognosy.)

Punicalagin, a natural polyphenolic compound classified as an ellagitannin, is a major ingredient of pomegranate (Punica granatum L.). Punicalagin has potent antioxidant and anti-inflammatory effects. Although the antinociceptive effects of orally administered pomegranate extracts have been reported, little is known about the effect of punicalagin on nociceptive transmission in the central nervous system. We examined whether punicalagin ameliorates neuropathic pain and inflammatory pain in the spinal cord. Male Sprague-Dawley rats were subjected to chronic constriction injury (CCI) of the sciatic nerve, and an intrathecal catheter was implanted for drug administration. The electronic von Frey test and cold-plate test were performed in CCI rats to evaluate mechanical and cold hyperalgesia in neuropathic pain, and the formalin test was performed in normal rats to evaluate acute and persistent inflammatory pain. An open-field test was conducted to explore whether punicalagin affects locomotor activity in CCI rats. Punicalagin administered intrathecally attenuated mechanical and cold hyperalgesia to the same degree as gabapentin in CCI rats and reduced pain-related behaviors in both the early and late phases in formalin-injected rats. Punicalagin did not affect motor function. These results suggest that punicalagin exerts an antinociceptive effect in the spinal cord without motor deficit, thus showing therapeutic potential for neuropathic pain and inflammatory pain., (© 2021. The Japanese Society of Pharmacognosy.)

Recent advances in cryo-electron microscopy (cryo-EM) have enabled protein structure determination at atomic resolutions. Cryo-EM specimens are prepared by rapidly freezing a protein solution on a metal grid coated with a holey carbon film; this results in the formation of an ice film on each hole. The thickness of the ice film is a critical factor for high-resolution structure determination; ice that is too thick degrades the contrast of the protein image while ice that is too thin excludes the protein from the hole or denatures the protein. Therefore, trained researchers need to manually select "good" regions with appropriate ice thicknesses for imaging. To reduce the time spent on such tasks, we developed a deep learning program consisting of a "detector" and a "classifier" to identify good regions from low-magnification EM images. In our method, the holes in a low-magnification EM image are detected via a detector, and the ice image on each hole is classified as either good or bad via a classifier. The detector detected more than 95% of the holes regardless of the type of samples. The classifier was trained for different types of samples because the appropriate ice thickness varies between sample types. The accuracies of the classifiers were 93.8% for a soluble protein sample (β-galactosidase) and 95.3% for a membrane protein sample (bovine heart cytochrome c oxidase). In addition, we found that a training data set containing ~ 2100 hole images from 300 low-magnification EM images was sufficient to obtain good accuracy, such as higher than 90%. We expect that the throughput of the cryo-EM data collection step will be greatly improved by using our method.

Respiratory rate, a sensitive indicator of respiratory status, is rarely measured during the field walking test. Our objective was to develop and validate a non-invasive, wearable monitoring system using stretchable strain sensors and an accompanying algorithm capable of providing real-time measurements of respiration during exercise. Twenty-four healthy volunteers wore stretchable sensors during a walking test protocol that included standing, sitting, walking, and walking with a stick. Sensors were placed on the ribcage and abdomen. The Bland-Altman method was used to assess the accuracy and precision of breath counts; total respiration time and inspiration time ratio were determined by custom algorithms and compared with measurements obtained with the standard flow sensor. The output signal from the stretchable sensor was highly synchronized with flow signals. The limits of agreement were within 3 breaths/min throughout the test protocol. Differences between sensors for total respiration time and inspiration time ratio were less than 14% and 26%, respectively. The agreement was maintained regardless of respiratory rate or volume. The wearable respiratory monitoring system yielded accurate and precise breath counts and total duration of respiratory cycle during moderate exercise in healthy young individuals, suggesting that it might be useful in clinical practice. Graphical abstract.

Background: The lack of high-quality clinical evidences hindered broad consensus on optimal therapies for primary nephrotic syndromes. The aim of the present study was to compare prevalence of immunosuppressive drug use in patients with primary nephrotic syndrome across 6 regions in Japan., Methods: Between 2009 and 2010, 380 patients with primary nephrotic syndrome in 56 hospitals were enrolled in a prospective cohort study [Japan Nephrotic Syndrome Cohort Study (JNSCS)], including 141, 151, and 38 adult patients with minimal change disease (MCD), membranous nephropathy (MN), and focal segmental glomerulosclerosis (FSGS), respectively. Their clinical characteristics were compared with those of patients registered in a large nationwide registry of kidney biopsies [Japan Renal Biopsy Registry (J-RBR)]. The regional prevalence of use of each immunosuppressive drug was assessed among adult MCD, MN, and FSGS patients who underwent immunosuppressive therapy in the JNSCS (n = 139, 127, and 34, respectively). Predictors of its use were identified using multivariable-adjusted logistic regression models., Results: The clinical characteristics of JNSCS patients were comparable to those of J-RBR patients, suggesting that the JNSCS included the representatives in the J-RBR. The secondary major immunosuppressive drugs were intravenous methylprednisolone [n = 33 (24.6%), 24 (19.7%), and 9 (28.1%) in MCD, MN, and FSGS, respectively] and cyclosporine [n = 25 (18.7%), 62 (50.8%), and 16 (50.0%), respectively]. The region was identified as a significant predictor of use of intravenous methylprednisolone in MCD and MN patients., Conclusion: Use of intravenous methylprednisolone for MCD and MN differed geographically in Japan. Its efficacy should be further evaluated in a well-designed trial.

Estimated continuous cardiac output (esCCO), a noninvasive technique for continuously measuring cardiac output (CO), is based on modified pulse wave transit time, which in turn is determined by pulse oximetry and electrocardiography. However, its trending ability has never been evaluated in patients undergoing non-cardiac surgery. Therefore, this study examined esCCO's ability to detect the exact changes in CO, compared with currently available arterial waveform analysis methods, in patients undergoing kidney transplantation. CO was measured using an esCCO system and arterial pressure-based CO (APCO), and compared with a corresponding intermittent bolus thermodilution CO (ICO) method. Percentage error and statistical methods, including concordance analysis and polar plot analysis, were used to analyze results from 15 adult patients. The difference in the CO values between esCCO and ICO was -0.39 ± 1.15 L min(-1) (percentage error, 35.6 %). And corrected precision for repeated measures was 1.16 L min(-1) (percentage error for repeated measures, 36.0 %). A concordance analysis showed that the concordance rate was 93.1 %. The mean angular bias was -1.8° and the radial limits of agreement were ±37.6°. The difference between the APCO and ICO CO values was 0.04 ± 1.37 L min(-1) (percentage error, 42.4 %). And corrected precision for repeated measures was 1.37 L min(-1) (percentage error for repeated measures, 42.5 %). The concordance rate was 89.7 %, with a mean angular bias of -3.3° and radial limits of agreement of ±42.2°. This study demonstrated that the trending ability of the esCCO system is not clinically acceptable, as judged by polar plots analysis; however, its trending ability is clinically acceptable based on a concordance analysis, and is comparable with currently available arterial waveform analysis methods.

Purpose: Our aim was to determine whether the maximum standardized uptake value (SUVmax) of the primary lesion demonstrated by [(18)F]-fluoro-2-deoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) is associated with the prognosis of maxillary sinus cancer., Materials and Methods: The relationships of clinicopathological factors including age, T stage, N stage, histologic type, treatment strategy, and primary tumor SUVmax with progression-free (PFS) and overall (OS) survival were evaluated using the log-rank test and Cox method in 31 patients with maxillary sinus cancer before combined superselective intra-arterial chemotherapy using high-dose cisplatin with concurrent radiotherapy, or radiotherapy alone., Results: The median duration of follow-up was 55.4 (range 9.7-72.6) months. PFS and OS of patients exhibiting a high SUVmax (≥16 and ≥17, respectively) for the primary tumor were significantly lower than those of patients for whom the primary tumor SUVmax was low (p = 0.0010 and p = 0.033, respectively). Multivariate analyses showed that T stage (p = 0.0049) and primary tumor SUVmax (p = 0.026) were independently prognostic of poorer PFS and that only primary tumor SUVmax (p = 0.049) was independently prognostic of poorer OS., Conclusion: SUVmax of the primary tumor determined by FDG-PET/CT before treatment could be a good surrogate marker for prognostication of maxillary sinus cancer.

Background: A closed-cell stent called Enterprise has been used for stent-assisted coil embolization of cerebral aneurysms. The Enterprise stent tends to cause kinks and vessel wall malposition in curved vessels and may cause thromboembolic complications. We evaluated vessel wall apposition of a new closed-cell stent, Enterprise 2, compared with a previous Enterprise stent, using curved vascular silicone models., Methods: The Enterprise or Enterprise 2 stent was deployed in curved vascular models with various radii of approximately 5 to 10 mm. Stent deployment was performed 25 times in each stent. A push-pull technique was used to minimize incomplete wall apposition. To evaluate conformity of stents, gaps between a stent and a vessel wall were measured., Results: The gap ratio (gap / a wall diameter) was 15 % ± 17 % (mean ± standard deviation) and 41 % ± 15 % with the Enterprise 2 stent and the Enterprise stent, respectively. Taking gap ratios and radii of vessel curvature into consideration, the Enterprise 2 stent had significantly better wall apposition than the Enterprise stent (p = 0.005). In the same radius of vessel curvature, the Enterprise 2 stent had approximately half of the gap compared with the Enterprise stent. There were no significant differences in vessel straightening effects between the two stents., Conclusions: The Enterprise 2 stent has better wall apposition in curved vessels than the Enterprise stent. The gap between a vessel wall and the Enterprise 2 stent is approximately half that of the Enterprise stent. However, gaps and kinks are still present in curved vessels with a small radius. Caution should be taken for kinks and malposition in acutely curved vessels, such as the siphon of the internal carotid artery.

The aim of the study was to investigate nerve fibers (NF) in human fetal livers. An immunohistochemical study was performed. NF were classified into portal tract innervation (PoI) and parenchymal innervation (PaI). The hilum area showed many Pol NF at 7 GW, and NF increased with gestational week (GW). Direct innervations to biliary epithelium were recognized. In large portal tracts, a few NCAM-positive mesenchymal cells were seen at 8 GW and many mesenchymal cells were noted around 12 GW. Apparent NF emerged around 15 GW, and NF increased with GW. Many NF plexuses were seen in 30-40 GW. In small portal tracts, no NF were seen in 7-10 GW. A few NCAM-positive mesenchymal cells emerged in 11 GW, and they increased thereafter. Apparent NF were seen around 20 GW and NF increased with GW. At term (40 GW), PoI NF were still immature. Ductal plate (DP) was positive for NCAM, NSE, chromogranin and synaptophysin, and direct innervations to DP were seen. The direct innervations to developing bile ducts and peribiliary glands were also seen. PaI NF were first seen at 21 GW and was consistent until 40 GW in which a few NF were seen in PaI. These observations suggest that PoI NF arise from committed portal mesenchyme. PaI NF are very immature at 40 GW. There are direct innervations to bile ducts, peribiliary glands, portal veins, hepatic arteries, and DP.

Background: Flow patterns in cerebral aneurysms are clinically important. Information on inflow patterns into aneurysms is especially helpful in preventing a recurrence after coil embolization. Computational fluid dynamics (CFD) simulations of patient-specific cerebral aneurysms are feasible and provide information on flow patterns. However, flow visualization by CFD simulations is challenging for recurrent aneurysms after coil embolization because coils make it difficult to obtain precise geometry of the recurrent aneurysms. In this study, we assessed the feasibility of flow visualization of recurrent aneurysms using 3D phase-contrast magnetic resonance imaging (PC-MRI)., Method: Time-of-flight magnetic resonance angiography and 3D PC-MRI were performed in eight cases of recurrent aneurysms after coil embolization. We attempted to visualize flow inside the aneurysms using data of 3D PC-MRI and evaluated the visualization. Additionally, CFD simulations were performed in a single case., Results: Inflow into aneurysms was visualized in all eight cases (100%). Flow patterns inside aneurysms were visualized in six cases (75%), and these were associated with a large size of recurrent aneurysms (mean size, 10.3 mm for visualized cases vs. 4.8 mm for unvisualized cases; p = 0.046, Mann-Whitney test). Flow patterns were similar between PC-MRI and CFD simulations. PC-MRI was faster and easier for observing inflow patterns than CFD simulations., Conclusions: This is the first study to demonstrate that flow visualization of recurrent aneurysms by 3D PC-MRI is feasible. This technique may be more practical and easier than CFD simulations, and may provide clinically helpful information.

Background: Carotid artery stenting (CAS) requires follow-up imaging to assess in-stent restenosis (ISR). This study aimed to determine whether non-enhanced magnetic resonance angiography (NE-MRA) is useful for evaluating ISR., Method: Between 2009 and 2013, we performed 118 consecutive CAS procedures using the Precise stent (n = 78) and the Carotid Wallstent (n = 40). We reviewed 1.5 T NE-MRA and examined visualization of the stent lumen and the degree of ISR if present. Other imaging modalities were used as references., Results: NE-MRA performed just after CAS was not able to visualize the stent lumen in all patients because of metal artifacts. In the Carotid Wallstent group, follow-up NE-MRA was available in 22 patients. The stent lumen was visible more than three months after CAS in all patients. Among them, >40 % ISR was observed by other modalities in eight lesions. The degree of restenosis measured by NE-MRA (y%) had a linear relationship with that measured by conventional angiography (x%) (y = 0.97x-0.4, r = 0.79, P = 0.021). In one case among 17 without ISR (6 %), NE-MRA showed false ISR. In the Precise stent group, NE-MRA did not visualize the stent lumen in the follow-up period., Conclusions: NE-MRA can visualize the stent lumen in the Carotid Wallstent more than three months after CAS, but not in the Precise stent at follow-up. This delayed visualization might depend on endothelialization of the stent lumen. The degree of ISR measured by NE-MRA is comparable to that by conventional angiography. NE-MRA can evaluate ISR after CAS with the Carotid Wallstent (100 % sensitivity and 94 % specificity).

Evaluation of the estimated continuous cardiac output (esCCO) allows non-invasive and continuous assessment of cardiac output. However, the applicability of this approach in children has not been assessed thus far. We compared the correlation coefficient, bias, standard deviation (SD), and the lower and upper 95 % limits of agreement for esCCO and dye densitography-cardiac output (DDG-CO) measurements by pulse dye densitometry (PDD) in adults and children. On the basis of these assessments, we aimed to examine whether esCCO can be used in pediatric patients. DDG-CO was measured by pulse dye densitometry (PDD) using indocyanine green. Modified-pulse wave transit time, obtained using pulse oximetry and electrocardiography, was used to measure esCCO. Correlations between DDG-CO and esCCO in adults and children were analyzed using regression analysis with the least squares method. Differences between the two correlation coefficients were statistically analyzed using a correlation coefficient test. Bland-Altman plots were used to evaluate bias and SD for DDG-CO and esCCO in both adults and children, and 95 % limits of agreement (bias ± 1.96 SD) and percentage error (1.96 SD/mean DDG-CO) were calculated and compared. The average age of the adult patients (n = 10) was 39.3 ± 12.1 years, while the average age of the pediatric patients (n = 7) was 9.4 ± 3.1 years (p < 0.001). For adults, the correlation coefficient was 0.756; bias, -0.258 L/min; SD, 1.583 L/min; lower and upper 95 % limits of agreement for DDG-CO and esCCO, -3.360 and 2.844 L/min, respectively; and percentage error, 42.7 %. For children, the corresponding values were 0.904; -0.270; 0.908; -2.051 and 1.510 L/min, respectively; and 35.7 %. Due to the high percentage error values, we could not establish a correlation between esCCO and DDG-CO. However, the 95 % limits of agreement and percentage error were better in children than in adults. Due to the high percentage error, we could not confirm a correlation between esCCO and DDG-CO. However, the agreement between esCCO and DDG-CO seems to be higher in children than in adults. These results suggest that esCCO can also be used in children. Future studies with bigger study populations will be required to further investigate these conclusions.

Background: In Y-stent-assisted coil embolization for cerebral aneurysms, open or closed cell stents are used. Different microcatheters for coil insertion are available. We investigated which microcatheter could be navigated into an aneurysm through a Y-stent with different stents., Methods: Double Neuroform open-cell stents or double Enterprise closed-cell stents were deployed in Y-configuration in a silicon model of a bifurcation aneurysm. Two endovascular neurosurgeons independently tried to navigate an SL-10 microcatheter for 0.010" coils or a PX Slim microcatheter for 0.020" Penumbra coils into the aneurysm through the Y-stent. In addition, we measured lengths of stent pores of the Y-stents with double Enterprise stents deployed in the model by micro-computed tomography., Results: It was feasible to navigate an SL-10 microcatheter into the aneurysm through the Y-stent with Enterprise or Neuroform stents. Navigation of a PX Slim microcatheter was feasible in the Y-stents only with Neuroform stents. In the Y-stent with double Enterprise stents, the lengths of the second stent pores were significantly smaller than those of the first stent (0.41 ± 0.18 mm vs 0.69 ± 0.20 mm; P = 0.008). The SL-10 microcatheter was smaller than approximately 80% of the stent pores of the first stent and 30% of those of the second stent. The PX Slim microcatheter was smaller than 20% of the stent pores of the first stent and 0% of those of the second stent., Conclusions: It was feasible to insert an SL-10 microcatheter into the aneurysm through Y-stents with Enterprise or Neuroform stents. Navigation of a PX Slim microcatheter for 0.020" Penumbra coils was feasible in Y-stents with Neuroform stents, but not with double Enterprise stents. The measurements of stent pores by micro-computed tomography supported this feasibility study. These results may be helpful to select appropriate stents and microcatheters in Y-stent-assisted coil embolization, especially in case of retreatments.

Background: Carotid artery stenting (CAS) is generally performed via a transfemoral approach. A transbrachial approach is usually chosen as an alternative when CAS via a transfemoral approach is difficult. At our institutions, a transcervical approach is chosen when the previous two approach routes are not available. We reviewed CAS cases treated via the transcervical route in our 1,067 CAS series to investigate the safety, feasibility, and frequency of this procedure as an approach route of CAS., Methods: We performed 1,067 CAS procedures in 1,067 consecutive cases between December 2002 and June 2011. Initially, a transfemoral route was chosen, and secondarily a transbrachial route, the last choice was a transcervical route. A transbrachial approach was chosen in 96 (9.0%) cases and a transcervical approach in ten (0.9%). We reviewed the characteristics and outcomes of CAS performed via a transcervical approach., Results: CAS was successfully performed on all ten transcervical-approach patients. Eight procedures were performed under local anesthesia and two under general anesthesia. Perioperative morbidity and mortality were both 0%. The modified Rankin scale (mRS) showed no deterioration at 3 months except for one case whose mRS became five because of an embolic stroke after aortic valve replacement., Conclusions: CAS via a transcervical approach was safe and feasible, and its frequency chosen as an approach route was 0.9%. This procedure can be an alternative to transfemoral or transbrachial approaches when CAS via either of these approaches is too difficult.

Background: Dual antiplatelet therapy for stent-assisted coiling of cerebral aneurysms is essential to prevent thromboembolic complications. There is concern that Y-stent-assisted coiling may increase thromboembolic complications compared with coiling with a single stent. Several reports have demonstrated that cilostazol may improve clopidogrel responsiveness. We investigated whether triple antiplatelet therapy with addition of cilostazol to aspirin plus clopidogrel for Y-stents can prevent thromboembolic events., Methods: Between July 2010 and October 2012, we treated 40 consecutive aneurysms with coil embolization using Enterprise stents. At the peri-procedural period, dual antiplatelet agents (100 mg aspirin and 75 mg clopidogrel) were used for the single stent group (n = 36), and triple antiplatelet agents (addition of 200 mg cilostazol) were used for the Y-stent group (n = 4). We evaluated post-operative diffusion-weighted imaging (DWI) and any complications. We assessed the following for statistical analysis: age, sex, aneurysm location, shape, and size, neck size, size of parent vessels, and stent length., Results: We found two neurological peri-procedural complications: one transient ischemic attack and one infarction. Both complications belonged to the Y-stent group, which was a significant factor of thromboembolic events (P = 0.008). There were no other significant factors related to neurological complications or positive DWI. For subgroup analysis of the single stent group, stent length was significantly longer in positive DWI than negative DWI (P = 0.04). In the follow-up period of 20 ± 8.6 months, there were no symptomatic late complications in any patients., Conclusions: Although the number of patients in the Y-stent group is small, this group had a significantly higher risk of thromboembolic complications. While our protocol of a routine dose of dual antiplatelet therapy may be sufficient for single stent therapy, our protocol of a routine dose of triple antiplatelet therapy for Y-stents may not prevent thromboembolic events. This suggests that evaluation of platelet function may be essential, especially for Y-stents.

Computational fluid dynamics (CFD) studies on cerebral aneurysms have attempted to identify surrogate hemodynamic parameters to predict rupture risk. We present a case of bilateral mirror image aneurysms, one of which ruptured soon after imaging. Wall shear stress values of the ruptured aneurysm changed by 20-30% after rupture because of change in the aneurysm shape. Findings from our case suggest that CFD studies comparing unruptured and ruptured aneurysms may not yield valid estimation on aneurysm rupture risk because of changes in aneurysm shape after rupture. Changes in aneurysm shape after rupture should be considered in CFD research.

The expression of NCAM (CD56), chromogranin A, synaptophysin, c-KIT (CD117) and PDGFRA in normal non-neoplastic skin and basal cell carcinoma (BCC) has rarely been investigated. The author immunohistochemically examined the expression of these molecules in 66 consecutive cases of BCC. In non-tumorous skin, NCAM chromogranin A, synaptophysin, c-KIT and PDGFRA expression was seen in the basal cell of the epidermis. NCAM, c-KIT and PDGFRA expression was also seen in the sweat glands and outer cells of hair follicles, but chromogranin and synaptophysin expression was not identified in these structures. In BCC, NCAM expression was seen in 95 % (63/66 cases). Its expression was membranous. Chromogranin A expression was recognized in 27 % (18/66 cases). Its expression was cytoplasmic. Synaptophysin expression was seen in 18 % (12/66 cases). Its expression was membranous and cytoplasmic. c-KIT expression was noted in 93 % (61/66 cases). Its expression was membranous and focally cytoplasmic. PDGFRA expression was seen in 65 % (43/66 cases). Its expression was membranous and cytoplasmic. The expression of these molecules in normal non-tumorous skin is a new finding. The expression of c-KIT and PDGFRA in BCC is also a new finding. In conclusion, the author described the normal (non-neoplastic) distribution of NCAM, chromogranin A, synaptophysin, c-KIT and PDGFRA. In addition, the author showed that, in cutaneous BCC, the expression of NCAM and c-KIT was high (95 and 93 % respectively), PDGFRA was intermediate (65 %), and chromogranin A and synaptophysin was relatively low (27 and 18 %, respectively).

Although several case series of primary signet-ring cell adenocarcinoma (SRCA) of the urinary bladder (UB) have been published, primary pure SRCA of the UB composed exclusively of signet-ring cells is rare. In addition, there have been no comprehensive studies of immunohistochemical studies of primary SRCA of the UB. This report shows immunohistochemical profiles of 3 cases of primary SRCA of UB. The age of patients was 37, 43, and 84 years. The male-to-female ratio was 2:1. One specimen was cystectomy, and other two specimens were transurethral bladder tumorectomy. Two patients died of bladder tumor 12 and 16 months after the diagnosis, and one patient is now alive 6 months after the diagnosis. Histologically, all the three tumors were composed exclusively of SRCA cells. Histochemically, all tumors contained neutral and acidic mucins. Immunohistochemically, consistent findings were positive cytokeratin (CK) AE1/3, CK CAM5.2, CK8, CK18, CK19, CK20, p53, Ki-67 (30, 50, 70%), MUC1, and MUC2. CK34BE12, CK5/6, CK14, EMA, p63, vimentin, and MUC6 were consistently negative. CK7 was positive in 1/3, CA19-9 1/3, and MUC5AC 2/3 cases. These data may provide basic knowledge of SRCA of the UB. In conclusion, the author presents three cases of primary pure SRCA with an emphasis of immunohistochemical findings.

Background: Human serum albumin is composed of human mercaptoalbumin (HMA) with cysteine residues having reducing powers and of oxidized human non-mercaptoalbumin. Previously, we reported that a lower HMA level is closely related to serious cardiovascular disease (CVD) incidence and mortality among hemodialysis patients. However, the relationship between HMA level and CVD incidence among peritoneal dialysis (PD) patients is unclear., Methods: We measured the redox state of human serum albumin using high-performance liquid chromatography in 30 continuous ambulatory PD patients. The association between HMA and incidental CVD events was evaluated., Results: Eight patients experienced symptomatic CVD events (5 patients died) at the 5-year follow-up. The concentration and fraction of HMA (cHMA and f(HMA), respectively) showed significantly lower values in patients with CVD than those without CVD (cHMA 1.58 ± 0.39 and 2.16 ± 0.43 g/dL, f(HMA) 48.9 ± 5.4 and 56.4 ± 8.6%, respectively). Multiple forward stepwise regression analysis using cHMA and f(HMA) as the criterion variables was performed, and C-reactive protein and hemoglobin were adopted as significant explanatory variables in the former equation, whereas urea nitrogen was adopted in the latter equation. Multiple logistic regression analysis revealed that cHMA is a statistically, and f(HMA) is a marginally significant explanatory variable of CVD incidence (p = 0.0369, R = -0.260 and p = 0.0580, R = -0.214, respectively)., Conclusions: Lower HMA level, which might be caused by chronic inflammation, anemia and accumulation of dialyzable uremic toxin(s), is closely related to serious CVD incidence among PD patients.

A 64-year-old woman presented with left occipital headache and right dissociated sensory loss due to hematomyelia on the left ventral side of C1 caused by rupture of an aneurysm on one of the feeders extending from the anterior spinal artery to complex epidural or dural and intradural arteriovenous fistulas (AVFs). Branches from the left occipital and ascending pharyngeal arteries and those from the left C2 radicular, left posterior spinal and anterior spinal arteries formed these multiple shunts, linking with a common venous drain flowing into the right petrosal vein. Surgical interception of all the shunts was achieved, making it unnecessary to directly treat the aneurysm in the spinal cord. The feeders, aneurysm and AVFs were not visualized on postoperative angiography, and the patient returned to a normal working life.

The author investigated histopathology of malignant changes of gastric foveolar hyperplastic polyps (GFHP). A total of 497 GFHP from 412 patients were retrospectively examined. Malignant changes were present in 11 GHP (2.2%). They were focal malignancies and well-differentiated adenocarcinoma without apparent invasion. In all the 11 GFHP, dysplastic glands were present in the vicinity of carcinomatous foci. Focal intestinal metaplasia was recognized in one case and was absent in the remaining 10 GFHP. Focal dysplastic glands were recognized in 51 GFHP (10%). Of these, four GFHP were associated with focal intestinal metaplasia, but the remaining 47 GFHP were not associated with intestinal metaplasia. Intestinal metaplasia alone was recognized in 23 GFHP (5%). Immunohistochemically, all carcinomatous foci within the 11 GFHP with malignant transformation showed positive p53 expression and high Ki-67 labeling. Of the 51 GFHP with dysplasia, 42 GFHP were positive for p53 protein, and the remaining 9 GFHP were negative for p53 protein. The dysplastic lesions of the 51 GFHP showed relatively high Ki-67 index. Intestinal metaplasia within GFHP was negative for p53 protein and showed low Ki-67 labeling. These results suggest that malignant transformation of GFHP may occur in 2.2% of cases, and that malignant changes develop via hyperplasia-dysplasia-carcinoma sequence in GFHP. Intestinal metaplasia within GFHP was not associated with carcinogenesis of GFHP.

Primary clear cell adenocarcinoma of the lung is extremely rare. A 63-year-old Japanese man consulted to our hospital because of cough and sputum. Imaging modalities including XP, CT and MRI revealed a tumor of the right middle lobe. They did not identify other tumors in the body. Because clinical cytology and biopsy showed malignant cells, segmentectomy of the lobe was performed. Grossly, the tumor was whitish tan tumor measuring 5 × 6 × 5 cm. Histologically, the tumor was composed entirely of clear cells arranged in papillary and tubular patterns. The tumor cells had hyperchromatic nuclei, and mitotic figure and nuclear stratification were scattered. Histochemically, glycogen and mucins were absent in tumor cell cytoplasm. Immunohistochemically, the tumor cells were positive for pancytokeratin (AE1/3, CAM5.2), cytokeratin (CK) 7, CK 8, CK18, CK19, EMA, CEA, CA19-9, CA125, p53, Ki-67 (labeling = 80%), TTF-1 and surfactant apoprotein A. In contrast, the tumor cells were negative for CK5/6, CK 34βE12, CK 14, CK 20, vimentin, desmin, S100 protein, α-smooth muscle actin, AFP, HMB45, CD10, CD34, HER2/neu, CD56, p63, and synaptophysin. The tumor recurred 6 months after the operation, and right middle lobectomy was performed. Postoperative imaging modalities showed no tumors. The patient is alive free from tumor 9 years after the first manifestation.

Neuroendocrine carcinoma of the esophagus including small-cell carcinoma and large-cell carcinoma is a very rare entity. The author herein reports a neuroendocrine carcinoma of the esophagus with an emphasis on KIT and PDGFRA. An 81-year-old man presented as dysphagia. An endoscopic examination revealed an ulcerated large tumor in the middle esophagus. A biopsy of the esophageal tumor identified an undifferentiated carcinoma consisting of malignant cells with hyperchromatic nuclei without differentiations. An immunohistochemical study revealed that the tumor cells were positive for cytokeratins, neuron-specific enolase, synaptophysin, p53 protein, KIT, and platelet-derived growth factor receptor-α (PDGFRA). The Ki-67 labeling was 100%. A pathologic diagnosis of neuroendocrine carcinoma with expression of KIT and PDGFRA was made. A genetic analysis using PCR direct sequencing method identified no mutations of KIT (exons 9, 11, 13, and 17) and PDGFRA (exons 12 and 18) genes. The patient underwent radiation and cisplatin-based chemotherapy. However, he developed systemic metastases and died of respiratory failure 8 months after the first manifestation. The present case is the first one of esophageal neuroendocrine carcinoma with an examination of protein expressions and gene mutations of KIT and PDGFRA.

The patient was a 30-year-old-pregnant woman (para 1, gradiva 1) without in utero exposure to diethylstilbestrol. A papillary and flat lesion was found by colposcopy in the uterine cervix in the routine examination at 10 pregnancy weeks. Biopsies showed papillary and tubular proliferation of malignant cells with clear glycogen-rich cytoplasm and malignant hobnail cells and were diagnosed as clear cell adenocarcinoma. The patient was treated by radical hysterectomy. The lesion of the cervix (10 × 15 × 5 mm) was clear cell adenocarcinoma with mild invasion into shallow myometrium (pTIb). An immunohistochemical study showed that the tumor cells were positive for pancytokeratins (AE1/3, CAM5.2, KL-1, and polyclonal wide), cytokeratin (CK) 7, CK8, CK18, CK19, epithelial membrane antigen, CA 125, p53 protein, and Ki-67 antigen (labeling = 95%). The tumor cells were negative for high-molecular weight CK (34βE12), CK5/6, CK14, CK20, vimentin, AFP, CEA, desmin, S100 protein, α-smooth muscle antigen, CD34, estrogen receptor, progesterone receptor, chromogranin, synaptophysin, neuron-specific enolase, and CD56. The patient is now of free of tumor. In the English literature, the present case is the second case of cervical clear cell adenocarcinoma during pregnancy, and the first case of cervical clear cell carcinoma investigating a wide range of immunoreactive antigens.

Primary lymphoma of the heart is extremely rare and almost always occurs in immunodeficient persons. The author reports a very rare autopsy case of primary diffuse large B-cell lymphoma in an immunocompetent woman. A 55-year-old woman presented with chest pain and arrhythmia. Various imaging modalities including US, CT, MRI, and gallium-scintigraphy revealed a mass in the right atrium and pericardial effusion. No tumor formations were recognized in other organs. Pericardial effusion cytology revealed malignant large lymphoid cells of B-cell type positive for CD20. Laboratory data did not reveal immunosuppression, and human immunodeficiency virus was negative. Therefore, a clinical diagnosis of primary heart lymphoma was made. The patient was treated with chemo-radiation, but died of heart failure 4 years after the first presentation. An autopsy revealed a tumor formation (4 × 5 × 6 cm) in the right atrium and ventricle of the heart. No tumor formation was recognized in other organs. Histological and immunohistochemical studies showed that the cardiac tumor was diffuse large B-cell lymphoma of non-germinal center B-cell type; tumor cells were positive for CD45, CD20, CD79α, bcl-2, and λ-chain, but negative for EBV latent membrane antigen 1, CD10, bcl-6 and MUM1. Epstein-Barr virus protein and DNA were negative by immunohistochemistry and in situ hybridization. It was suggested that primary lymphoma of the heart may occur in immunocompetent persons. This is the first report of diffuse large B-cell lymphoma of the heart with a classification of non- or germinal center B-cell type.

An extremely rare case of primary small cell carcinoma of the pleura with an emphasis on KIT and platelet-derived growth factor receptor-α (PDGFRA) genes is reported here. A 67-year-old man underwent left testicular orchiectomy because of a testicular tumor. The tumor was immunohistochemically shown to be diffuse large B-cell lymphoma. The patient was treated with chemotherapy and radiation and followed up in our hospital. Eight years after the orchiectomy, the patient (75 years old) developed left pleural tumor and pleural effusion, and a biopsy was performed. The biopsy revealed a medullary malignant tumor consisting of small round and spindle cells. The following three possibilities were considered: recurrent lymphoma, mesothelioma, and small cell carcinoma. Immunohistochemically, the tumor cells were positive for cytokeratin, synaptophysin, CD56, KIT, and PDGFRA, but negative for CEA, cytokeratin 5/6, neuron-specific enolase, chromogranin, CD45, CD3, CD20, CD45RO, CD15, CD30, calretinin, WT-1, B72.3, D2-40, and TTF-1. Therefore, a diagnosis of pleural small cell carcinoma was made. A molecular genetic analysis using PCR-direct sequencing identified no mutations of KIT (exons 9, 11, 13, and 17) and PDGFRA (exons 12 and 18) genes. The patient was treated with chemotherapy (cisplatin) and radiation (50 Gray). The present case is the first reported case of primary small cell carcinoma of the pleura with an examination of KIT and PDGFRA expressions and KIT and PDGFRA gene mutations.

The author reports a rare case of sarcomatoid carcinoma with an emphasis on immunohistochemical features. A 79-year-old man was admitted to our hospital because of hematuria. An endoscopy revealed a large polypoid tumor in the bladder, and urine cytology demonstrated malignant cells. A cystectomy was performed. The patient is now alive without metastasis 4 months after the operation. Grossly, a large polypoid tumor (5 x 6 x 5 cm) was present in the bladder. Microscopically, the tumor consisted of high-grade transitional cell carcinoma element (10% in area) and sarcomatoid element (90% in area). There was a gradual transition between the two. The tumor cells were invaded into peribladder tissue (pT3b). Immunohistochemically, the sarcomatoid element was positive for four types of pancytokeratins, high-molecular weight cytokeratin (CK), CK5/6, CK7, CK18, CK19, epithelial membrane antigen (EMA), vimentin, p53 protein, p63, Ki-67 (labeling = 92%), neuron-specific enolase (NSE), and platelet-derived growth factor receptor-alpha (PDGFRA). It was negative for CK14, CK20, melanosome, carcinoembryonic antigen (CEA), desmin, S100 protein, myoglobin, alpha-smooth muscle antigen (ASMA), CD34, chromogranin, synaptophysin, CD56, CD68, and KIT. The transitional cell carcinoma element showed similar immunoreactivity except for negative CK5/6, positive CK20, and negative vimentin. A molecular genetic analysis of KIT gene (exons 9, 11, 13, and 17) and PDGFRA (exons 12 and 18) gene with the use of PCR-direct sequencing showed no mutations. The present case is the first report of sarcomatoid carcinoma of the urinary bladder demonstrating extensive immunohistochemistry and mutational status of KIT and PDGFRA genes. The sarcomatoid carcinoma in the present case may be derived from sarcomatous differentiation of high-grade transitional cell carcinoma.

Background: Oxidative stress is enhanced in advanced chronic kidney disease (CKD) patients and recognized as a main contributor to cardiovascular disease. Carbonyl stress is also known to be enhanced in advanced CKD; however the precise relationship between oxidative stress and carbonyl stress is not clear. The aim of this study was to investigate potential relationships between oxidative stress, carbonyl stress, and renal function among predialysis patients with CKD., Methods: A total of 32 predialysis CKD patients (22 male, 10 female) were divided into four groups according to their values for creatinine clearance (Ccr) (group A, > or =60 ml/min; group B, 45-59 ml/min; group C, 30-44 ml/min; group D, < or =29 ml/min). As main markers of oxidative and carbonyl stresses, the redox state of Cys-34 (free thiol group) of human serum albumin [HSA(Cys-34)-redox] and the carbonyl content of serum proteins were employed, respectively., Results: The values for the fraction of both reversibly oxidized HSA [f(HNA-1)] and irreversibly oxidized HSA [f(HNA-2)] significantly increased with a decrease in renal function (group A, 21.0 +/- 3.4 and 1.8 +/- 0.3%; group D, 31.1 +/- 4.1 and 2.7 +/- 0.9%, respectively). The value for carbonyl content also significantly increased with a decrease in renal function (group A, 0.7 +/- 0.1 nmol/mg protein; group D, 1.1 +/- 0.2 nmol/mg protein). There was a significant positive correlation between carbonyl content and the f(HNA-2) value, while such a correlation was not observed between carbonyl content and the f(HNA-1) value, suggesting that there is a close relationship between serum protein carbonylation and irreversible albumin thiol oxidation., Conclusions: There is a close relationship between oxidative stress and carbonyl stress and these are enhanced in correlation with the level of renal dysfunction among predialysis CKD patients.

The author reports a rare case of primary large cystic extragastrointestinal stromal tumor (eGIST) of the transverse mesocolon with genetic analyses of the c-kit and platelet-derived growth factor receptor-alpha (PDGFRA) genes. A 78-year-old man was found to have a large cystic tumor in the abdomen, and the tumor was resected. Grossly, the tumor was located in the transverse mesocolon, and cystic. Microscopically, the tumor consisted of epithelioid cells with atypia. Mitotic figures were noted in five of 50 high power fields. Immunohistochemically, the tumor cells were positive for KIT, CD34, PDGFRA, and vimentin, but negative for cytokeratins, neuron specific enolase, desmin, S100 protein, alpha-smooth muscle actin, p53 protein, HMB45, CD68, CEA, factor VIII-related antigen, chromogranin, and synaptophysin. Ki67 labeling was 5%. Genetically, the tumor showed a point mutation (GAC --> GTC) at codon 842 of exon 18 of the PDGFRA gene. Exon 12 of the PDGFRA gene and exons 9, 11, 13, and 17 of the c-kit gene showed no mutations. No recurrence is noted 3 years after the operation. This case shows that eGIST may occur in the transverse mesocolon.

The author reports herein an extremely rare case of primary small cell carcinoma of the mediastinum with an emphasis on KIT and PDGFRA genes. A 66-year-old man was found to have a mediastinal tumor on a routine chest X-ray examination, and was admitted to our hospital. Imaging modalities revealed a 5 x 4 cm tumor in the middle mediastinum near the bronchial carina. No other tumors were detected in the body including the lungs. Video-assisted thoracoscopy confirmed the mediastinal tumor, and a large incisional biopsy was performed. The tumor was histologically small cell carcinoma. An immunohistochemical study revealed positive reactions for cytokeratins (AE1/3, polyclonal), synaptophysin, neuron-specific enolase, CD56, KIT, and PDGFRA, and negative reactions for chromogranin, CEA, CD45, CD20, and CD3. Ki-67 labeling showed a value of 80%. A molecular genetic analysis using PCR-direct sequencing identified no mutations of KIT (exons 9, 11, 13, and 17) and PDGFRA (exons 12 and 18) genes. The patient received radiation and chemotherapy, and the tumor was fully resolved. The patient has remained free of recurrence for 6 years after the first presentation. The present case is the first reported case of primary small cell carcinoma of the mediastinum with an examination of KIT and PDGFRA expressions and KIT and PDFGRA gene mutations.

Monomeric carbonyl reductases (CBRs) are enzymes that catalyze the reduction of many endogenous and xenobiotic carbonyl compounds, including steroids and prostaglandins. There are two monomeric CBR genes in the human genome, cbr1 and cbr3, which exhibit high homology in their amino acid sequences. Human CBR1 (hCBR1) is known as prostaglandin 9-keto reductase and 15-hydroxy dehydrogenase, and regulates the metastasis of cancer cells through the regulation of prostaglandin metabolism. However, there is little information concerning the molecular and enzymatic characteristics of human CBR3 (hCBR3). The present study demonstrated the tissue and cellular localization, and catalytic activity of hCBR3. Semi-quantitative PCR revealed the ubiquitous but lower expression of hCBR3 compared with that of hCBR1. Bacterially expressed hCBR3 exhibited limited catalytic activity toward menadione, 4-benzoylpyridine, and 4-nitrobenzaldehyde. Similar results were obtained when the cell lysates of CBR-overexpressing HEK293 cells were examined. Additionally, neither the prostaglandin 9-keto reductase nor the 15-hydroxy dehydrogenase activities of hCBR3 were significant. Immunofluorescence staining revealed that ectopically expressed hCBR3 proteins were localized in the cytosol of HEK293 cells. These results suggested that hCBR3 and hCBR1 play distinct physiological roles. This study expands our understanding of the relationship between the two monomeric hCBRs and prostaglandin metabolism.

'Lymphomatosis cerebri' (LC) is a term indicating a diffusely infiltrating form of primary central nervous system lymphoma (PCNSL) without evidence of a mass lesion. Not infrequently, diagnostic confusion is caused by its presentation on cranial magnetic resonance images (MRI), which is characterized by diffuse leukoencephalopathy without contrast enhancement. In this report, we describe a 53-year-old, immunocompetent man who had an insidiously progressive dementia and right weakness. On serial MRI in 4 months duration, diffuse white matter lesions without contrast enhancement gradually progressed, which was clinically consistent with his worsening condition. Biopsy specimen demonstrated nondestructive, diffusely infiltrating, large B-cell lymphoma, diagnosing LC. After the biopsy, corticosteroids were initiated, which dramatically alleviated his symptoms. Afterwards, he was treated by whole brain irradiation (total 36Gy) and discharged without noticeable deficits. Diagnosis of LC requires additional examinations generally not performed in the other white matter disorders. In suspected cases, biopsy should be performed to avoid deferring adequate cytostatic treatment.

H(+)-coupled peptide transporter 1 (PEPT1) and the basolateral peptide transporter mediate the absorption of small peptides and peptide-like drugs in the small intestine. Recently, we constructed a mathematical model to simulate glycylsarcosine (Gly-Sar) transport in Caco-2 cells. In this study, we attempted to adjust our model to a change in the expression level of PEPT1. To obtain cell lines expressing PEPT1 at different levels, recloning of Caco-2 cells was performed, and nine clones were isolated. Compared with parental cells, clones 1 and 9 exhibited the lowest and the highest levels of [(14)C]Gly-Sar uptake from the apical side, respectively, whereas activities of the basolateral peptide transporter were comparable. Kinetic analysis demonstrated that the difference in the activity of PEPT1 was accounted by variations in V (max). Moreover, PEPT1 mRNA level was positively related to the activity of [(14)C]Gly-Sar uptake (r=0.55). Based on these findings, the V (max) value of PEPT1 was defined as a variable using the amount of PEPT1 mRNA as an index of the expression level. With this improved model, Gly-Sar transport in clones 1 and 9 was well-predicted, suggesting that our model can simulate Gly-Sar transport in cells expressing PEPT1 at different levels.

Small peptides and some pharmacologically active compounds are absorbed from the small intestine by the apical H(+)-coupled peptide transporter 1 (PEPT1) and the basolateral peptide transporter. Here we investigated the efflux properties of the basolateral peptide transporter in Caco-2 cells using two strategies, efflux measurements and a kinetic analysis of transepithelial transport of glycylsarcosine (Gly-Sar). [(14)C]Gly-Sar efflux through the basolateral membrane was not affected significantly by the external pH. Both approaches revealed that the basolateral peptide transporter was saturable in the efflux direction, and that the affinity was lower than that in the influx direction. For two peptide-like drugs, there was no difference in substrate recognition by the basolateral peptide transporter between the two sides of the membrane. Using the kinetic parameters of PEPT1 and the basolateral peptide transporter, a computational model of Gly-Sar transport in Caco-2 cells was constructed. The simulation fitted the experimental data well. Our findings suggested that substrate affinity of the basolateral peptide transporter is apparently asymmetric, but pH-dependence and substrate specificity are symmetric for the two directions of transport. The behaviour of Gly-Sar in Caco-2 cells could be predicted by a mathematical model describing the peptide transporters.