Background and Purpose
To investigate the diagnostic performance of diffusion-weighted imaging (DWI) and contrast-enhanced imaging in combination with T2-weighted imaging (T2WI) for magnetic resonance imaging (MRI) evaluation of intrapelvic recurrence of gynecological malignancies.
Materials and Methods
Sixty-two patients with suspected intrapelvic recurrence of gynecological malignancies underwent pelvic MRI including T2WI DWI, and contrast-enhanced imaging. Diagnostic performance for detection of local recurrence, pelvic lymph node and bone metastases, and peritoneal lesions was evaluated by consensus reading of two experienced radiologists using a 5-point scoring system, and compared among T2WI with unenhanced T1-weighted imaging (T1WI) (protocol A), a combination of protocol A and DWI (protocol B), and a combination of protocol B and contrast-enhanced imaging (protocol C). Final diagnoses were obtained by histopathological examinations, radiological imaging and clinical follow-up for at least 6 months. Receiver operating characteristic (ROC) analysis and McNemar test were employed for statistical analysis.
Locally recurrent disease, lymph node recurrence, peritoneal dissemination and bone metastases were present in 48.4%, 29.0%, 16.1%, and 6.5% of the patients, respectively. The patient-based sensitivity, specificity, accuracy, and area under the ROC curve (AUC) for detection of intrapelvic recurrence were 55.0, 81.8, 64.5% and 0.753 for protocol A, 80.0, 77.3, 79.0% and 0.838 for protocol B, and 80.0, 90.9, 83.9% and 0.862 for protocol C, respectively. The sensitivity, accuracy, and AUC were significantly better for protocols B and C than for protocol A (p<0.001). There was no significant difference between protocols B and C.
MRI using a combination of DWI and T2WI gives comparatively acceptable results for assessment of intrapelvic recurrence of gynecological malignancies.
Recent studies on the large Maf transcription factors have shown that Mafb and Mafa have respective and distinctive roles in β-cell development and maturation. However, whether this difference in roles is due to the timing of the gene expression (roughly, expression of Mafb before birth and of Mafa after birth) or to the specific function of each gene is unclear. Our aim was to examine the functional differences between these genes that are closely related to β cells by using an in vivo model of β-like cell generation. We monitored insulin gene transcription by measuring bioluminescence emitted from the liver of insulin promoter-luciferase transgenic (MIP-Luc-VU) mice. Adenoviral gene transfers of Pdx1/Neurod/Mafa (PDA) and Pdx1/Neurod/Mafb (PDB) combinations generated intense luminescence from the liver that lasted for more than 1 week and peaked at 3 days after transduction. The peak signal intensities of PDA and PDB were comparable. However, PDA but not PDB transfer resulted in significant bioluminescence on day 10, suggesting that Mafa has a more sustainable role in insulin gene activation than does Mafb. Both PDA and PDB transfers ameliorated the glucose levels in a streptozotocin (STZ)-induced diabetic model for up to 21 days and 7 days, respectively. Furthermore, PDA transfer induced several gene expressions necessary for glucose sensing and insulin secretion in the liver on day 9. However, a glucose tolerance test and liver perfusion experiment did not show glucose-stimulated insulin secretion from intrahepatic β-like cells. These results demonstrate that bioluminescence imaging in MIP-Luc-VU mice provides a noninvasive means of detecting β-like cells in the liver. They also show that Mafa has a markedly intense and sustained role in β-like cell production in comparison with Mafb.
Mutually exclusive expression of T-bet and Eomes drives the development of distinct NK cell lineages with complementary functions.
Trail+DX5−Eomes− natural killer (NK) cells arise in the mouse fetal liver and persist in the adult liver. Their relationships with Trail−DX5+ NK cells remain controversial. We generated a novel Eomes-GFP reporter murine model to address this question. We found that Eomes− NK cells are not precursors of classical Eomes+ NK cells but rather constitute a distinct lineage of innate lymphoid cells. Eomes− NK cells are strictly dependent on both T-bet and IL-15, similarly to NKT cells. We observed that, in the liver, expression of T-bet in progenitors represses Eomes expression and the development of Eomes+ NK cells. Reciprocally, the bone marrow (BM) microenvironment restricts T-bet expression in developing NK cells. Ectopic expression of T-bet forces the development of Eomes− NK cells, demonstrating that repression of T-bet is essential for the development of Eomes+ NK cells. Gene profile analyses show that Eomes− NK cells share part of their transcriptional program with NKT cells, including genes involved in liver homing and NK cell receptors. Moreover, Eomes− NK cells produce a broad range of cytokines, including IL-2 and TNF in vitro and in vivo, during immune responses against vaccinia virus. Thus, mutually exclusive expression of T-bet and Eomes drives the development of different NK cell lineages with complementary functions.
Despite convincing evidence that 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)—a heterocyclic amine generated by cooking meats at high temperatures—is carcinogenic in animal models, it remains unclear whether PhIP exposure leads to increased cancer risk in humans. PhIP-DNA adduct levels were measured in specimens from 534 prostate cancer case-control pairs nested within a historical cohort of men with histopathologically benign prostate specimens. We estimated the overall and race-stratified risk of subsequent prostate cancer associated with higher adduct levels. PhIP-DNA adduct levels in benign prostate were significantly higher in Whites than African Americans (0.274 Optical Density Units (OD) ±0.059 vs. 0.256 OD ±0.054; p<0.0001). Prostate cancer risk for men in the highest quartile of PhIP-DNA adduct levels was modestly increased (Odds Ratio (OR) = 1.25; 95% confidence interval (CI) = 0.76-2.07). In subset analyses, the highest risk estimates were observed in White patients diagnosed more than 4 years after cohort entry (OR=2.74; 95% CI=1.01-7.42) or under age 65 (OR=2.80; 95% CI=0.87-8.97). In Whites, cancer risk associated with high grade prostatic intraepithelial neoplasia combined with elevated PhIP-DNA adduct levels (OR=3.89; 95% CI=1.56-9.73) was greater than risk associated with either factor alone. Overall, elevated levels of PhIP-DNA adducts do not significantly increase prostate cancer risk. However, our data show that White men have higher PhIP-DNA adduct levels in benign prostate tissue than African American men, and suggest that in certain subgroups of White men high PhIP-DNA adduct levels may predispose to an increased risk for prostate cancer.
dna adducts; nested case-control study; immunohistochemistry; carcinogens; imidazoles; biopsy, needle
The postnatal proliferation and maturation of insulin-secreting pancreatic β-cells are critical for glucose metabolism and disease development in adults. Elucidation of the molecular mechanisms underlying these events will be beneficial to direct the differentiation of stem cells into functional β-cells. Maturation of β-cells is accompanied by increased expression of MafA, an insulin gene transcription factor. Transcriptome analysis of MafA knockout islets revealed MafA is required for the expression of several molecules critical for β-cell function, including Glut2, ZnT8, Granuphilin, Vdr, Pcsk1 and Urocortin 3, as well as Prolactin receptor (Prlr) and its downstream target Cyclin D2 (Ccnd2). Inhibition of MafA expression in mouse islets or β-cell lines resulted in reduced expression of Prlr and Ccnd2, and MafA transactivated the Prlr promoter. Stimulation of β-cells by prolactin resulted in the phosphorylation and translocation of Stat5B and an increased nuclear pool of Ccnd2 via Prlr and Jak2. Consistent with these results, the loss of MafA resulted in impaired proliferation of β-cells at 4 weeks of age. These results suggest that MafA regulates the postnatal proliferation of β-cells via prolactin signaling.
Paroxysmal kinesigenic dyskinesia is characterized by sudden attacks of involuntary movements. It is often misdiagnosed clinically as psychogenic illness, which distresses the patients to a great extent. A correct diagnosis will improve the quality of life in patients with paroxysmal kinesigenic dyskinesia because treatment with low doses of anticonvulsants is effective for eliminating the clinical manifestations. Paroxysmal kinesigenic dyskinesia can occur independently of or concurrently with benign infantile convulsion. Identification of PRRT2 as the causative gene of benign infantile convulsion and paroxysmal kinesigenic dyskinesia allows genetic confirmation of the clinical diagnosis.
We describe the clinical features of a Japanese family with either paroxysmal kinesigenic dyskinesia or benign infantile convulsion. A PRRT2 missense mutation (c.981C > G, p.Ile327Met) was identified in two patients with benign infantile convulsion and three patients with paroxysmal kinesigenic dyskinesia as well as in two unaffected individuals. Allowing incomplete penetrance in the mutation carriers, this mutation co-segregated completely with the phenotype. The patients with paroxysmal kinesigenic dyskinesia had been misdiagnosed with psychogenic illness for many years. They were correctly diagnosed with paroxysmal kinesigenic dyskinesia when their children visited a pediatrician for benign infantile convulsion. Treatment with carbamazepine controlled their involuntary movements completely.
Paroxysmal kinesigenic dyskinesia is a treatable movement disorder that is often misdiagnosed clinically as psychogenic illness. It is important to note that two clinically distinct disorders, benign infantile convulsion and paroxysmal kinesigenic dyskinesia, are allelic conditions caused by PRRT2 mutations. Paroxysmal kinesigenic dyskinesia should be suspected in families with a child with benign infantile convulsion.
Benign infantile convulsion; Mutation; Paroxysmal kinesigenic dyskinesia; PRRT2; Seizures
Activation of peroxisome proliferator-activated receptor (PPAR) α disrupts growth-related activities in a variety of human cancers. This study was designed to determine whether fenofibrate, a PPARα agonist, can suppress 4-nitroquinoline 1-oxide (4-NQO)-induced proliferative lesions in the lung of obese hyperlipidemic mice. Male Tsumura Suzuki Obese Diabetic mice were subcutaneously injected with 4-NQO to induce lung proliferative lesions, including adenocarcinomas. They were then fed a diet containing 0.01% or 0.05% fenofibrate for 29 weeks, starting 1 week after 4-NQO administration. At week 30, the incidence and multiplicity (number of lesions/mouse) of pulmonary proliferative lesions were lower in mice treated with 4-NQO and both doses of fenofibrate compared with those in mice treated with 4-NQO alone. The incidence and multiplicity of lesions were significantly lower in mice treated with 4-NQO and 0.05% fenofibrate compared with those in mice treated with 4-NQO alone (p < 0.05). Both doses of fenofibrate significantly reduced the proliferative activity of the lesions in 4-NQO-treated mice (p < 0.05). Fenofibrate also significantly reduced the serum insulin and insulin-like growth factor (IGF)-1 levels, and decreased the immunohistochemical expression of IGF-1 receptor (IGF-1R), phosphorylated Akt, and phosphorylated Erk1/2 in lung adenocarcinomas. Our results indicate that fenofibrate can prevent the development of 4-NQO-induced proliferative lesions in the lung by modulating the insulin-IGF axis.
4-nitroquinoline-1-oxide; lung neoplasms; carcinogenesis; hyperlipidemia; hyperinsulinemia; chemoprevention
We have established a transgenic rat for adenocarcinoma of the prostate (TRAP) model that features uniform adenocarcinoma development in prostatic lobes at high incidence within a short experimental period. However, no invasive carcinomas with reactive stroma characteristics similar to those in man were observed. We therefore have focused on a new model for invasive carcinoma of the prostate using TRAP rats. In experiment 1, male TRAP rats in groups 1 and 2 were treated with orchiectomy at day 0 of the experiment. Rats in groups 1–3 underwent testosterone propionate (TP) implantation from weeks 1 to 4 and from weeks 6 to 16. Rats in groups 1 and 3 were given 3,2’-dimethyl-4-aminobiphenyl (DMAB) after TP implantation. The rats of group 4 served as controls. In experiment 2, the rats were divided into three groups, none of which received DMAB or orchiectomy, treated with TP continuously or with the treatment withdrawn once or twice. In experiment 1, invasive adenocarcinomas with abundant collagenous stroma were found in the dorsolateral and anterior prostate, some of which showed perineural space invasion at week 16. The number of invasive carcinoma foci was most frequent in group 3. In experiment 2, invasive adenocarcinoma development in the lateral prostates was correlated with the number of TP administration/withdrawal cycles. In conclusion, our newly established rat model for invasive adenocarcinoma of the prostate could serve as a useful preclinical model for evaluating the in vivo efficacy of preventive and therapeutic agents targeting of the tumor microenvironment.
prostate cancer; animal model; cancer invasion; transgenic rat; testosterone propionate; intermittent administration
Cyclin-dependent kinase 5 (Cdk5) is essential for brain development and function, and its deregulated expression is implicated in some of neurodegenerative diseases. We reported earlier that the forebrain-specific Cdk5 conditional knockout (cKO) mice displayed an early lethality associated with neuroinflammation, increased expression of the neuronal tissue-type plasminogen activator (tPA), and neuronal migration defects.
In order to suppress neuroinflammation in the cKO mice, we first treated these mice with pioglitazone, a PPARγ agonist, and analyzed its effects on neuronal loss and longevity. In a second approach, to delineate the precise role of tPA in neuroinflammation in these mice, we generated Cdk5 cKO; tPA double knockout (dKO) mice.
We found that pioglitazone treatment significantly reduced astrogliosis, microgliosis, neuronal loss and behavioral deficit in Cdk5 cKO mice. Interestingly, the dKO mice displayed a partial reversal in astrogliosis, but they still died at early age, suggesting that the increased expression of tPA in the cKO mice does not contribute significantly to the pathological process leading to neuroinflammation, neuronal loss and early lethality.
The suppression of neuroinflammation in Cdk5 cKO mice ameliorates gliosis and neuronal loss, thus suggesting the potential beneficial effects of the PPARγ agonist pioglitazone for the treatment for neurodegenerative diseases.
Neuroinflammation; Cdk5; Pioglitazone; tPA; Cdk5 conditional knockout mice
T helper (Th)2 cells have been proposed to play a neuroprotective role in multiple sclerosis (MS). This is mainly based on “loss-of-function” studies in an animal model for MS, experimental autoimmune encephalomyelitis (EAE), using blocking antibodies against Th2 related cytokines, and knockout mice lacking Th2-related molecules. We tested whether an increase of Th2 responses (“gain-of-function” approach) could alter EAE, the approach of novel GATA binding protein 3 (GATA3)-transgenic (tg) mice that overexpress GATA3, a transcription factor required for Th2 differentiation. In EAE induced with myelin oligodendrocyte glycoprotein (MOG)35–55 peptide, GATA3-tg mice had a significantly delayed onset of disease and a less severe maximum clinical score, compared with wild-type C57BL/6 mice. Histologically, GATA3-tg mice had decreased levels of meningitis and demyelination in the spinal cord, and anti-inflammatory cytokine profiles immunologically, however both groups developed similar levels of MOG-specific lymphoproliferative responses. During the early stage, we detected higher levels of interleukin (IL)-4 and IL-10, with MOG and mitogen stimulation of regional lymph node cells in GATA3-tg mice. During the late stage, only mitogen stimulation induced higher IL-4 and lower interferon-γ and IL-17 production in GATA3-tg mice. These results suggest that a preexisting bias toward a Th2 immune response may reduce the severity of inflammatory demyelinating diseases, including MS.
autoimmune demyelinating diseases; GATA3 transcription factor; autoimmunity; animal models; paraffin; histology; oligodendrocyte-myelin glycoprotein; Th1-Th2 assays; Luxol fast blue; Th17; incomplete Freund’s adjuvant
Ulcerative colitis (UC) is an inflammatory bowel disease, and its pathogenesis includes
genetic, environmental, and immunological factors, such as T helper cells and their
secreted cytokines. T helper cells are classified as Th1, Th2, and Th17 cells. However, it
is unclear which T helper cells are important in UC. Dextran sulfate sodium (DSS)-induced
colitis is a commonly used model of UC. In this study, we induced DSS colitis in Th1
dominant (T-bet transgenic (Tg)) mice, Th2 dominant (GATA-3 Tg) mice, and Th17 dominant
(RORγt Tg) mice to elucidate the roles of T helper cell in DSS colitis. The results showed
that GATA-3 Tg mice developed the most severe DSS colitis compared with the other groups.
GATA-3 Tg mice showed a significant decreased in weight from day 1 to day 7, and an
increased high score for the disease activity index compared with the other groups.
Furthermore, GATA-3 Tg mice developed many ulcers in the colon, and many neutrophils and
macrophages were detected on day 4 after DSS treatment. Measurement of GATA-3-induced
cytokines demonstrated that IL-13 was highly expressed in the colon from DSS-induced
GATA-3 Tg mice. In conclusion, GATA-3 overexpression in T-cells and IL-13 might play
important roles in the development of DSS colitis.
dextran sulfate sodium; GATA-3; IL-13; inflammatory bowel disease; T helper cell
Cre/loxP system-mediated site-specific recombination is utilized to study gene function
in vivo. Successful conditional knockout of genes of interest is
dependent on the availability of Cre-driver mice. We produced and characterized pancreatic
β cell-specific Cre-driver mice for use in diabetes mellitus research. The gene encoding
Cre was inserted into the second exon of mouse Ins1 in a bacterial
artificial chromosome (BAC). Five founder mice were produced by microinjection of
linearized BAC Ins1-cre. The transgene was integrated between
Mafa and the telomere on chromosome 15 in one of the founders, BAC
Ins1-cre25. To investigate Cre-loxP recombination, BAC Ins1-cre25 males were crossed with
two different Cre-reporters, R26R and R26GRR females. On gross observation, reporter
signal after Cre-loxP recombination was detected exclusively in the adult pancreatic
islets in both F1 mice. Immunohistological analysis indicated that Cre-loxP
recombination-mediated reporter signal was colocalized with insulin in pancreatic islet
cells of both F1 mice, but not with glucagon. Moreover, Cre-loxP recombination
signal was already observed in the pancreatic islets at E13.5 in both F1
fetuses. Finally, we investigated ectopic Cre-loxP recombination for
Ins1, because the ortholog Ins2 is also expressed in the
brain, in addition to the pancreas. However, there was no Cre-loxP recombination-mediated
reporter signal in the brain of both F1 mice. Our data suggest that BAC
Ins1-cre25 mice are a useful Cre-driver C57BL/6N for pancreatic β cell-specific Cre-loxP
recombination, except for crossing with knock-in mice carrying floxed gene on chromosome
cre-driver mice; cre-loxP recombination; diabetes; insulin1; pancreatic β cells
Fluorescent proteins with light wavelengths within the optical window are one of the
improvements in in vivo imaging techniques. Near-infrared (NIR)
fluorescent protein (iRFP) is a stable, nontoxic protein that emits fluorescence within
the NIR optical window without the addition of exogenous substrate. However, studies
utilizing an in vivo iRFP model have not yet been published. Here, we
report the generation of transgenic iRFP mice with ubiquitous NIR fluorescence expression.
iRFP expression was observed in approximately 50% of the offspring from a matings between
iRFP transgenic and WT mice. The serum and blood cell indices and body weights of iRFP
mice were similar to those of WT mice. Red fluorescence with an excitation wavelength of
690 nm and an emission wavelength of 713 nm was detected in both newborn and adult iRFP
mice. We also detected fluorescence emission in whole organs of the iRFP mice, including
the brain, heart, liver, kidney, spleen, lung, pancreas, bone, testis, thymus, and adipose
tissue. Therefore, iRFP transgenic mice may therefore be a useful tool for various types
of in vivo imaging.
fluorescent protein; in vivo imaging; iRFP; optical window
Orexin/hypocretin neurons have a crucial role in the regulation of sleep and wakefulness. To help determine how these neurons promote wakefulness, we generated transgenic mice in which orexin neurons expressed halorhodopsin (orexin/Halo mice), an orange light-activated neuronal silencer. Slice patch-clamp recordings of orexin neurons that expressed halorhodopsin demonstrated that orange light photic illumination immediately hyperpolarized membrane potential and inhibited orexin neuron discharge in proportion to illumination intensity. Acute silencing of orexin neurons in vivo during the day (the inactive period) induced synchronization of the electroencephalogram and a reduction in amplitude of the electromyogram that is characteristic of slow-wave sleep (SWS). In contrast, orexin neuron photoinhibition was ineffective during the night (active period). Acute photoinhibition of orexin neurons during the day in orexin/Halo mice also reduced discharge of neurons in an orexin terminal field, the dorsal raphe (DR) nucleus. However, serotonergic DR neurons exhibited normal discharge rates in mice lacking orexin neurons. Thus, although usually highly dependent on orexin neuronal activity, serotonergic DR neuronal activity can be regulated appropriately in the chronic absence of orexin input. Together, these results demonstrate that acute inhibition of orexin neurons results in time-of-day-dependent induction of SWS and in reduced firing rate of neurons in an efferent projection site thought to be involved in arousal state regulation. The results presented here advance our understanding of the role of orexin neurons in the regulation of sleep/wakefulness and may be relevant to the mechanisms that underlie symptom progression in narcolepsy.
To evaluate the diagnostic value of retrospective image fusion from pelvic magnetic resonance imaging (MRI) and 18F-fluorodeoxyglucose positron emission tomography (PET) in detecting intra-pelvic recurrence of gynecological tumor.
Thirty patients with a suspicion of recurrence of gynecological malignancy underwent inline contrast-enhanced PET/computed tomography (CT) and pelvic contrast-enhanced MRI for restaging. Diagnostic performance about the local recurrence, pelvic lymph node and bone metastasis and peritoneal lesion of PET/low-dose non-enhanced CT (PET/ldCT), PET/full-dose contrast-enhanced CT (PET/ceCT), contrast-enhanced MRI, and retrospective image fusion from PET and MRI (fused PET/MRI) were evaluated by two experienced readers. Final diagnoses were obtained by histopathological examinations, radiological imaging and clinical follow-up for at least 6 months. McNemar test was employed for statistical analysis.
Documented positive locally recurrent disease, pelvic lymph node and bone metastases, and peritoneal dissemination were present in 53.3, 26.7, 10.0, and 16.7 %, respectively. Patient-based sensitivity for detecting local recurrence, pelvic lymph node and bone metastasis and peritoneal lesion were 87.5, 87.5, 100 and 80.0 %, respectively, for fused PET/MRI, 87.5, 62.5, 66.7 and 60.0 %, respectively, for contrast-enhanced MRI, 62.5, 87.5, 66.7 and 80.0 %, respectively, for PET/ceCT, and 50.0, 87.5, 66.7 and 60.0 %, respectively, for PET/ldCT. The sensitivity of diagnosing local recurrence by fused PET/MRI was significantly better than that of PET/ldCT (p = 0.041). The patient-based sensitivity, specificity and accuracy for the detection of intra-pelvic recurrence/metastasis were 91.3, 100 and 93.3 % for fused PET/MRI, 82.6, 100 and 86.7 % for contrast-enhanced MRI, 82.6, 100 and 86.7 % for PET/ceCT and 78.3, 85.7 and 80.0 % for PET/ldCT.
Fused PET/MRI combines the individual advantages of MRI and PET, and is a valuable technique for assessment of intra-pelvic recurrence of gynecological cancers.
Fused PET/MRI; PET/CT; MRI; Restaging; Gynecological tumor
Myoepithelial carcinoma (malignant myoepithelioma) of the breast is a rare tumor, for which only a limited number of reports have been published. Most of the reports emphasized diagnosis and pathology but not biological behavior and treatment. We report a 61-year-old patient with breast myoepithelial carcinoma who developed locoregional and distant metastases and received many chemotherapy regimens. She presented with an elastic hard mass of the left breast. Breast conserving surgery was performed as part of both diagnosis and treatment. From the results of histological and immunohistochemical examinations, this case was considered to be a myoepithelial carcinoma. Fifteen months after the completion of adjuvant radiotherapy, distant metastasis of the left parasternal lymph node metastasis developed. She was treated by further excision and received a total of four regimens of chemotherapy including a combination of doxorubicin and cyclophosphamide. She received chemotherapy for 20 months after the diagnosis of metastasis.
Embryogenesis in placental mammals is sustained by exquisite interplay between the embryo proper and placenta. UTF1 is a developmentally regulated gene expressed in both cell lineages. Here, we analyzed the consequence of loss of the UTF1 gene during mouse development. We found that homozygous UTF1 mutant newborn mice were significantly smaller than wild-type or heterozygous mutant mice, suggesting that placental insufficiency caused by the loss of UTF1 expression in extra-embryonic ectodermal cells at least in part contributed to this phenotype. We also found that the effects of loss of UTF1 expression in embryonic stem cells on their pluripotency were very subtle. Genome structure and sequence comparisons revealed that the UTF1 gene exists only in placental mammals. Our analyses of a family of genes with homology to UTF1 revealed a possible mechanism by which placental mammals have evolved the UTF1 genes.
To evaluate the efficacy of using 3-tesla (T) magnetic resonance imaging (MRI) diagnosis of extracapsular extension (ECE) for decision-making about neurovascular bundle (NVB) preservation in robot-assisted radical prostatectomy (RARP) for prostate cancer (PC).
Materials and Methods
We prospectively collected data on PC patients (n=67) who underwent preoperative 3-T MRI before RARP. The choice between nerve sparing or resection was based on 3-T MRI findings of ECE. We compared the MRI findings with the pathological data on surgical margins. Our clinical staging in this study was defined only by MRI.
When the data were divided by prostate lobe (right lobe or left lobe, n=134), 3-T MRI showed 28 positive cases of ECE in 134 prostate lobes, allowing NVB preservation in 42 cases (31.3%). Nerve-sparing surgery was achieved in 38.7% of cases in which clinical T2 staging by MRI was reported. The pathological data revealed that 10 of 134 prostate lobes had positive ECE. The overall sensitivity, specificity, positive predictive value, and negative predictive value for predicting stage T3 (positive ECE) by side were 60.0% (12 of 20 sides), 86.0% (98 of 114 sides), 42.9% (12 of 28 sides), and 92.5% (98 of 106 sides), respectively.
Three-T MRI prior to RARP enables the use of ECE diagnosis to guide decision-making about NVB preservation, with comparatively high specificity and negative predictive value. Further prospective studies are underway to reach more definitive conclusions.
Diagnoses; Magnetic resonance imaging; Prostatectomy; Robotics
Introduction. The biological role of amyloid precursor protein (APP) is not well understood, especially in testicular germ cell tumors (TGCTs). Therefore, we aimed to investigate the immunoreactivity (IR) and expression of APP in TGCTs and evaluated its clinical relevance. Materials and Methods. We performed an analysis of immunohistochemistry and mRNA expression of APP in 64 testicular specimens and 21 snap-frozen samples obtained from 1985 to 2004. We then evaluated the association between APP expression and clinicopathological status in TGCTs. Results. Positive APP IR was observed in 9.8% (4/41) of seminomatous germ cell tumors (SGCTs) and 39.1% (9/23) of nonseminomatous germ cell tumors (NGCTs). NGCTs showed significantly more cases of positive IR (P = 0.00870) and a higher mRNA expression level compared with those of SGCTs (P = 0.0140). Positive APP IR was also significantly associated with α-fetoprotein (αFP) elevation (P = 0.00870) and venous invasion (P = 0.0414). Conclusion. We observed an elevated APP expression in TGCTs, especially in NGCTs. APP may be associated with a more aggressive cancer in TGCTs.
In diabetes research, bioluminescence imaging (BLI) has been applied in studies of β-cell impairment, development, and islet transplantation. To develop a mouse model that enables noninvasive imaging of β cells, we generated a bacterial artificial chromosome (BAC) transgenic mouse in which a mouse 200-kbp genomic fragment comprising the insulin I gene drives luciferase expression (Ins1-luc BAC transgenic mouse). BLI of mice was performed using the IVIS Spectrum system after intraperitoneal injection of luciferin, and the bioluminescence signal from the pancreatic region analyzed. When compared with MIP-Luc-VU mice [FVB/N-Tg(Ins1-luc)VUPwrs/J] expressing luciferase under the control of the 9.2-kbp mouse insulin I promoter (MIP), the bioluminescence emission from Ins1-luc BAC transgenic mice was enhanced approximately 4-fold. Streptozotocin-treated Ins1-luc BAC transgenic mice developed severe diabetes concomitant with a sharp decline in the BLI signal intensity in the pancreas. Conversely, mice fed a high-fat diet for 8 weeks showed an increase in the signal, reflecting a decrease or increase in the β-cell mass. Although the bioluminescence intensity of the islets correlated well with the number of isolated islets in vitro, the intensity obtained from a living mouse in vivo did not necessarily reflect an absolute quantification of the β-cell mass under pathological conditions. On the other hand, adenovirus-mediated gene transduction of β-cell-related transcription factors in Ins1-luc BAC transgenic mice generated luminescence from the hepatic region for more than 1 week. These results demonstrate that BLI in Ins1-luc BAC transgenic mice provides a noninvasive method of imaging islet β cells and extrapancreatic activity of the insulin gene in the liver under normal and pathological conditions.
Systemic lupus erythematosus is considered to be under the control of polygenic inheritance, developing according to the cumulative effects of susceptibility genes with polymorphic alleles; however, the mechanisms underlying the roles of polygenes based on functional and pathological genomics remain uncharacterized. In this study, we substantiate that a CD72 polymorphism in the membrane-distal extracellular domain impacts on both the development of glomerulonephritis and vasculitis in a lupus model strain of mice, MRL/MpJ-Faslpr, and the reactivity of BCR signal stimulation. We generated mice carrying a bacterial artificial chromosome transgene originating from C57BL/6 (B6) mice that contains the Cd72b locus (Cd72B6 transgenic [tg]) or the modified Cd72b locus with an MRL-derived Cd72c allele at the polymorphic region corresponding to the membrane-distal extracellular domain (Cd72B6/MRL tg). Cd72B6 tg mice, but not Cd72B6/MRL tg mice, showed a significant reduction in mortality following a marked improvement of disease associated with decreased serum levels of IgG3 and anti-dsDNA Abs. The number of splenic CD4−CD8− T cells in Cd72B6 tg mice was decreased significantly in association with a reduced response to B cell receptor signaling. These results indicate that the Cd72 polymorphism affects susceptibility to lupus phenotypes and that novel functional rescue by a bacterial artificial chromosome transgenesis is an efficient approach with wide applications for conducting a genomic analysis of polygene diseases.
The Cre/loxP system is a strategy for controlling temporal and/or spatial gene expression
through genome alteration in mice. As successful Cre/loxP genome alteration depends on
Cre-driver mice, Cre-reporter mice are essential for validation of Cre gene expression
in vivo. In most Cre-reporter mouse strains, although the presence of
reporter product indicates the expression of Cre recombinase, it has remained unclear
whether a lack of reporter signal indicates either no Cre recombinase expression or
insufficient reporter gene promoter activity. We produced a novel ROSA26 knock-in
Cre-reporter C57BL/6N strain exhibiting green emission before and red after Cre-mediated
recombination, designated as strain R26GRR. Ubiquitous green fluorescence and no red
fluorescence were observed in R26GRR mice. To investigate the activation of tdsRed,
EGFP-excised R26GRR, R26RR, mice were produced through the crossing of
C57BL/6N mice with R26GRR/Ayu1-Cre F1 mice. R26RR mice showed extraordinarily
strong red fluorescence in almost all tissues examined, suggesting ubiquitous activation
of the second reporter in all tissues after Cre/loxP recombination. Moreover, endothelial
cell lineage and pancreatic islet-specific expression of red fluorescence were detected in
R26GRR/Tie2-Cre F1 mice and R26GRR /Ins1-Cre F1 mice, respectively.
These results indicated that R26GRR mice are a useful novel Cre-reporter mouse strain. In
addition, R26GRR mice with a pure C57BL/6N background represent a valuable source of
green-to-red photoconvertible cells following Cre/loxP recombination for application in
transplantation studies. The R26GRR mouse strain will be available from RIKEN BioResource
CAG promoter; Cre-reporter mouse; EGFP; Rosa26; tdsRed
In humans, genetic variation and dietary factors may alter the biologic effects of exposure to 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), one of the major heterocyclic amines generated from cooking meats at high temperatures that has carcinogenic potential through the formation of DNA adducts. Previously, we reported grilled red meat consumption associated with PhIP-DNA adduct levels in human prostate. In the present study, we expanded our investigation to estimate the associations between beverage consumption and PhIP-DNA adduct levels in prostate for 391 prostate cancer cases. Of the 15 beverages analyzed, red wine consumption had the strongest association with PhIP-DNA adduct levels showing an inverse correlation in both tumor (p=0.006) and non-tumor (p=0.002) prostate cells. Red wine consumption differed significantly between African-American and white cases, but PhIP-DNA adduct levels in prostate did not vary by race. In African Americans compared with whites, however, associations between red wine consumption and PhIP-DNA adduct levels were not as strong as associations with specific (e.g., SULT1A1 and UGT1A10 genotypes) and non-specific (e.g., African ancestry) genetic variation. In a multivariable model, the covariate for red wine consumption explained a comparable percentage (13-16%) of the variation in PhIP-DNA adduct levels in prostate across the two racial groups, but the aforementioned genetic factors explained 33% of the PhIP-DNA adduct variation in African-American cases, while only 19% of the PhIPDNA adduct variation in whites. We conclude that red wine consumption may counteract biologic effects of PhIP exposure in human prostate, but genetic factors may play an even larger role, particularly in African Americans.
compounds, heterocyclic; resveratrol; UDP-glucuronosyltransferase; sulfotransferases; African Americans; chemoprevention
Cyclin-dependent kinase 5 (Cdk5) plays a pivotal role in neuronal migration and differentiation, and in axonal elongation. Although many studies have been conducted to analyze neuronal functions of Cdk5, its kinase activity has also been reported during oligodendrocyte differentiation, which suggests Cdk5 may play an important role in oligodendrocytes. Here, we describe a hypomyelination phenotype observed in Emx1-cre mediated Cdk5 conditional knockout (cKO) mice (Emx1-cKO), in which the Cdk5 gene was deleted in neurons, astrocytes and oligodendrocyte -lineage cells. In contrast, the Cdk5 gene in CaMKII cKO mice was deleted only in neurons. Because the development of mature oligodendrocytes from oligodendrocyte precursor cells is a complex process, we performed in situ hybridization using markers for the oligodendrocyte precursor cell and for the differentiated oligodendrocyte. Our results indicate that hypomyelination in Emx1-cKO is due to the impaired differentiation of oligodendrocytes, rather than to the proliferation or migration of their precursors. The present study confirmed the in vivo role of Cdk5 in oligodendrocyte differentiation.
Cyclin-dependent kinase 5; Oligodendrocyte differentiation; Hypomyelination; Conditional knockout mice
During vertebrate lens development, the anterior, ectoderm-derived lens vesicle cells differentiate into a monolayer of epithelial cells that retain proliferative potential. Subsequently, they exit the cell cycle and give rise to posterior lens fiber cells that form the lens body. In the present study, we demonstrate that the transcription factor GATA-3 is expressed in the posterior lens fiber cells during embryogenesis, and that GATA-3-deficiency impairs lens development. Interestingly, expression of E-cadherin, a premature lens vesicle marker, is abnormally prolonged in the posterior region of Gata3 homozygous mutant lenses. Furthermore, expression of γ-crystallin, a differentiation marker for fiber cells, is reduced. This suppressed differentiation is accompanied by an abnormal cellular proliferation, as well as with diminished levels of the cell-cycle inhibitors Cdkn1b/p27 and Cdkn1c/p57 and increased Ccnd2/cyclin D2 abundance. Thus, these observations suggest that GATA-3 is essential for lens cells differentiation and proper cell cycle control.
GATA-3; crystallin; lens fiber; differentiation; cell cycle; apoptosis