Prostaglandin E2 is a major soluble factor produced by stromal cells for preventing inflammatory cytokine production from dendritic cells

Dendritic cells (DCs) are specialized antigen-presenting cells that play pivotal roles in initiating immune responses. However, DC maturation is usually strongly restricted by the stromal microenvironment, especially in non-lymphoid tissues, such as skin and mucosa. Although suppression of DC maturation by stromal cells has been well documented, the molecular basis of this suppression has not been established. In this study, we examined the role of fibroblasts for DC maturation in vitro. The mouse embryonic fibroblasts (MEFs) strongly suppressed LPS-induced DC maturation. Although suppression of class II MHC and CD40 required DC-MEF contact, soluble factors in the culture supernatant of MEFs were sufficient for the suppression of IL-12 and tumor necrosis factor-alpha production. Using molecular-size selection and HPLC, we determined that prostaglandin E2 (PGE2) is a major soluble inhibitory factor secreted by MEFs. This was confirmed by the fact that cyclooxygenase inhibitors inhibited the production of the suppressive factor by MEFs. These results suggest that PGE2 is a major soluble factor produced by MEFs for the suppression of inflammatory cytokine production from DCs, while a contact mechanism between MEFs and DCs is required for the suppression to induce T cell-stimulating molecules.     

Computer-aided diagnosis for the classification of focal liver lesions by use of contrast-enhanced ultrasonography

The authors developed a computer-aided diagnostic (CAD) scheme for classifying focal liver lesions (FLLs) as liver metastasis, hemangioma, and three histologic differentiation types of hepatocellular carcinoma (HCC), by use of microflow imaging (MFI) of contrast-enhanced ultrasonography. One hundred and three FLLs obtained from 97 cases used in this study consisted of 26 metastases (15 hyper- and 11 hypovascularity types), 16 hemangiomas (five hyper- and 11 hypovascularity types) and 61 HCCs: 24 well differentiated (w-HCC), 28 moderately differentiated (m-HCC), and nine poorly differentiated (p-HCC). Pathologies of all cases were determined based on biopsy or surgical specimens. Locations and contours of FLLs on contrast-enhanced images were determined manually by an experienced physician. MFI was obtained with contrast-enhanced low-mechanical-index (MI) pulse subtraction imaging at a fixed plane which included a distinctive cross section of the FLL. In MFI, the inflow high signals in the plane, which were due to the vascular patterns and the contrast agent, were accumulated following flash scanning with a high-MI ultrasound exposure. In the initial step of our computerized scheme, a series of the MFI images was extracted from the original cine clip (AVI format). We applied a smoothing filter and time-sequential running average techniques in order to reduce signal noise on the single MFI image and cyclic noise on the sequential MFI images, respectively. A kidney, vessels, and a liver parenchyma region were segmented automatically by use of the last image of a series of MFI images. The authors estimated time-intensity curves for an FLL by use of a series of the temporally averaged MFI images in order to determine temporal features such as estimated replenishment times at early and delayed phases, flow rates, and peak times. In addition, they extracted morphologic and gray-level image features which were determined based on the physicians' knowledge of the diagnosis of the FLL, such as the size of lesion, vascular patterns, and the presence of hypoechoic regions. They employed a cascade of six independent artificial neural networks (ANNs) by use of extracted temporal and image features for classifying five types of liver diseases. A total of 16 temporal and image features, which were selected from 43 initially extracted features, were used for six different ANNs for making decisions at each decision in the cascade. The ANNs were trained and tested with a leave-one-lesion-out test method. The classification accuracies for the 103 FLLs were 88.5% for metastasis, 93.8% for hemangioma, and 86.9% for all HCCs. In addition, the classification accuracies for histologic differentiation types of HCCs were 79.2% for w-HCC, 50.0% for m-HCC, and 77.8% for p-HCC. The CAD scheme for classifying FLLs by use of the MFI on contrast-enhanced ultrasonography has the potential to improve the diagnostic accuracy in the histologic diagnosis of HCCs and the other liver diseases.     

Novel mutations of the GLA gene in Japanese patients with Fabry disease and their functional characterization by active site specific chaperone

Fabry disease is an X-linked recessive inborn metabolic disorder caused by a deficiency of the lysosomal enzyme alpha-galactosidase A (EC 3.2.1.22). The causative mutations are diverse, include both large rearrangements and single-base substitutions, and are dispersed throughout the 7 exons of the alpha-galactosidase A gene (GLA). Mutation hotspots for Fabry disease do not exist. We examined 62 Fabry patients in Japan and found 24 GLA mutations, including 11 novel ones. A potential treatment reported for Fabry disease is active site specific chaperone (ASSC) therapy using 1-deoxygalactonojirimycin (DGJ), an inhibitor of alpha-galactosidase A, at subinhibitory concentrations. We transfected COS-7 cells with the 24 mutant GLAs and analyzed the alpha-galactosidase A activities. We then treated the transfected COS-7 cells with DGJ and analyzed its effect on the mutant enzyme activities. The activity of 11 missense mutants increased significantly with DGJ. Although ASSC therapy is useful only for misfolding mutants and therefore not applicable to all cases, it may be useful for treating many Japanese patients with Fabry disease.     

Spatiotemporal recapitulation of central nervous system development by murine embryonic stem cell-derived neural stem/progenitor cells

Neural stem/progenitor cells (NS/PCs) can generate a wide variety of neural cells. However, their fates are generally restricted, depending on the time and location of NS/PC origin. Here we demonstrate that we can recapitulate the spatiotemporal regulation of central nervous system (CNS) development in vitro by using a neurosphere-based culture system of embryonic stem (ES) cell-derived NS/PCs. This ES cell-derived neurosphere system enables the efficient derivation of highly neurogenic fibroblast growth factor-responsive NS/PCs with early temporal identities and high cell-fate plasticity. Over repeated passages, these NS/PCs exhibit temporal progression, becoming epidermal growth factor-responsive gliogenic NS/PCs with late temporal identities; this change is accompanied by an alteration in the epigenetic status of the glial fibrillary acidic protein promoter, similar to that observed in the developing brain. Moreover, the rostrocaudal and dorsoventral spatial identities of the NS/PCs can be successfully regulated by sequential administration of several morphogens. These NS/PCs can differentiate into early-born projection neurons, including cholinergic, catecholaminergic, serotonergic, and motor neurons, that exhibit action potentials in vitro. Finally, these NS/PCs differentiate into neurons that form synaptic contacts with host neurons after their transplantation into wild-type and disease model animals. Thus, this culture system can be used to obtain specific neurons from ES cells, is a simple and powerful tool for investigating the underlying mechanisms of CNS development, and is applicable to regenerative treatment for neurological disorders.     

G-CSF-producing malignant pleural mesothelioma: an autopsy case report with literature review

This study reports a 54-year-old man who was a carpenter by occupation. He suffered from left chest and back pain and left pleural effusion. Peripheral blood showed granulocytosis and high serum titers of granulocyte-colony stimulating factor (G-CSF) and CYFRA. He died 20 months later. At autopsy, a pleural tumor located around the left lung and thickening of the pericardium, diaphragm, and esophagus by tumor infiltration was seen. The tumor proliferated in papillary and solid alveolar patterns by neoplastic cells. They were positive for calretinin, D2-40, CK5/6, HBME-1, G-CSF, CK19, and E-cadherin. He was diagnosed with G-CSF-producing epithelioid malignant pleural mesothelioma.     

Anococcygeal raphe revisited: a histological study using mid-term human fetuses and elderly cadavers

Purpose

We recently demonstrated the morphology of the anococcygeal ligament. As the anococcygeal ligament and raphe are often confused, the concept of the anococcygeal raphe needs to be re-examined from the perspective of fetal development, as well as in terms of adult morphology.

Materials and Methods

We examined the horizontal sections of 15 fetuses as well as adult histology. From cadavers, we obtained an almost cubic tissue mass containing the dorsal wall of the anorectum, the coccyx and the covering skin. Most sections were stained with hematoxylin and eosin or Masson-trichrome solution.

Results

The adult ligament contained both smooth and striated muscle fibers. A similar band-like structure was seen in fetuses, containing: 1) smooth muscle fibers originating from the longitudinal muscle coat of the anal canal and 2) striated muscle fibers from the external anal sphincter (EAS). However, in fetuses, the levator ani muscle did not attach to either the band or the coccyx. Along and around the anococcygeal ligament, we did not find any aponeurotic tissue with transversely oriented fibers connecting bilateral levator ani slings. Instead, in adults, a fibrous tissue mass was located at a gap between bilateral levator ani slings; this site corresponded to the dorsal side of the ligament and the EAS in the immediately deep side of the natal skin cleft.

Conclusion

We hypothesize that a classically described raphe corresponds to the specific subcutaneous tissue on the superficial or dorsal side of the anococcygeal ligament.     

Existence of a neuropathic pain component in patients with osteoarthritis of the knee

Purpose

Pain from osteoarthritis (OA) is generally classified as nociceptive (inflammatory). Animal models of knee OA have shown that sensory nerve fibers innervating the knee are significantly damaged with destruction of subchondral bone junction, and induce neuropathic pain (NP). Our objective was to examine NP in the knees of OA patients using painDETECT (an NP questionnaire) and to evaluate the relationship between NP, pain intensity, and stage of OA.

Materials and Methods

Ninety-two knee OA patients were evaluated in this study. Pain scores using Visual Analogue Scales (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), painDETECT, duration of symptoms, severity of OA using the Kellgren-Lawrence (KL) system, and amount of joint fluid were evaluated and compared using a Spearman''s correlation coefficient by rank test.

Results

Our study identified at least 5.4% of our knee OA patients as likely to have NP and 15.2% as possibly having NP. The painDETECT score was significantly correlated with the VAS and WOMAC pain severity. Compared with the painDETECT score, there was a tendency for positive correlation with the KL grade, and tendency for negative correlation with the existence and amount of joint fluid, but these correlations were not significant.

Conclusion

PainDETECT scores classified 5.4% of pain from knee OA as NP. NP tended to be seen in patients with less joint fluid and increased KL grade, both of which corresponded to late stages of OA. It is important to consider the existence of NP in the treatment of knee OA pain.     

A Pilot Study of the Water Quality of the Yarra River, Victoria, Australia, Using In Vitro Techniques

A pilot study was initiated to provide the first information on the recombinant receptor-reporter gene bioassay (hormonal) activity of freshwaters in Victoria. The project involved the collection of water samples from six stations on the main stem of the Yarra River in and upstream of the city of Melbourne, Australia in April 2008 and April 2009. Samples were prepared for measurement of sample toxicity using a modified photobacterium test, genotoxicity using a high-throughput luminescent umu test method, and human and medaka estrogen receptor (hERα and medERα), retinoic acid receptor (RAR), aryl hydrocarbon receptor (AhR) and thyroid receptor (TR) assay activity using the relevant yeast-based bioassays. Most samples were only weakly or moderately toxic, with no relationship observed to location along the river. The data for 2008 suggests that at that time the Yarra River samples contained few compounds that were, in and of themselves, genotoxic. No estrogenic or thyroid, and <1?ng/L retinoic acid receptor activity was observed. AhR activity increased with progressed downstream. AhR activity was higher in April 2009 than at the same time in 2008, perhaps as a result of extensive bush fires in the catchment in the months immediately prior to sampling. About 24% of the total AhR activity observed was associated with suspended solids.