Granulysin produced by uterine natural killer cells induces apoptosis of extravillous trophoblasts in spontaneous abortion

Immune changes are known to occur in recurrent spontaneous abortion, but it is unclear whether either maternal natural killer (NK) cells or T cells attack fetus-derived trophoblasts. To clarify the immunological causes of spontaneous abortion, we examined the relationship between cytotoxic granule proteins in decidual lymphocytes, such as granulysin, granzyme B, and perforin, and the induction of apoptosis in extravillous trophoblasts (EVTs). The number of granulysin-positive CD56^bright NK cells increased significantly in the decidua basalis during spontaneous abortion compared with normal pregnancy; however, granzyme B- and perforin-positive cells did not change. Interestingly, the expression of granulysin was also detected in the nuclei of EVTs in spontaneous abortion samples. When IL-2-stimulated CD56^bright NK cells were cocultured with EVT cells (HTR-8/SV40neo), granulysin was found initially in the cytoplasm and then accumulated in the nuclei of the HTR-8/SV40neo cells. Furthermore, transfected cells expressing a GFP-granulysin fusion protein induced apoptosis in HTR-8/SV40neo cells independently of caspases. Our results suggest that granulysin-positive uterine NK cells attack EVTs; subsequently, the uNK-derived granulysin actively accumulates in the nuclei of EVTs, causing the death of EVTs due to apoptosis. These data support a new apoptosis pathway for trophoblasts via uNK-derived granulysin, suggesting that granulysin is involved in spontaneous abortion.     

Comparison of MR imaging with CT in depiction of tumour extension into the pterygopalatine fossa.

The computed tomography (CT) and magnetic resonance imaging (MRI) results of 30 consecutive patients with tumours in the skull base, and who had abnormalities of the pterygopalatine fossa (PPF) on CT or MRI, were retrospectively compared with respect to visualization of tumour infiltration into the PPF. CT did not depict the abnormalities in the PPF in five patients (17%), while unenhanced T1-weighted MR images depicted tumour infiltration in all patients. Obliteration of PPF fat was better visualized on CT than T2-weighted and proton density weighted MR images, as were bony abnormalities. On MRI, intracranial extension was seen in eight of 25 patients with extracranial tumour. MRI is a sensitive method of demonstrating both tumour infiltration of the PPF and perineural tumour spread.     

Interleukin-8 and glucose in amniotic fluid, fetal fibronectin in vaginal secretions and preterm labor index based on clinical variables are optimal predictive markers for preterm delivery in patients with intact membranes

AIM: Various predictive markers for preterm delivery have been proposed in previous studies. We investigated which marker is most reliable. METHODS: In 126 patients with preterm labor before 32 weeks of gestation and intact membranes, who had regular uterine contractions with cervical changes effaced > or =50%, we evaluated seven markers: interleukin (IL)-8, glucose, and granulocyte count in amniotic fluid (AF); fetal fibronectin (fFN) in vaginal secretions; IL-8 in cervical mucus; cervical length; and preterm labor index (PLI) based on clinical variables. The relationships of these variables to the occurrence of preterm delivery before 34 weeks were examined by logistic regression analysis. RESULTS: Values for AF IL-8, AF granulocyte count, fFN in vaginal secretions, and PLI were significantly higher, while the value for AF glucose was significantly lower, in patients delivering before 34 weeks than those in patients delivering at or following 34 weeks (P < 0.0001 for all). The most sensitive marker for predicting delivery before 34 weeks was AF IL-8 (sensitivity, 67.8%). The most specific markers were AF IL-8 (specificity, 95.5%) and PLI (specificity, 95.5%). By the logistic regression analysis, AF IL-8, AF glucose, fFN in vaginal secretions and PLI showed independent relationships with delivery before 34 weeks (P = 0.0009, P = 0.0032, P = 0.0131 and P = 0.0038, respectively). CONCLUSIONS: In preterm labor with intact membranes, AF IL-8, AF glucose, fFN in vaginal secretions and PLI were highly predictive markers for detecting preterm delivery before 34 weeks.     

Sonographic femur length to trunk cross area ratio: prediction of fetal outcome in 30 cases in which micromelia was suspected

AIM: The purpose of the present study was to investigate the potential value of fetal routine sonographic biometry in evaluating micromelias. METHODS: Thirty fetuses had a presumptive diagnosis of micromelia from antepartum ultrasound examinations during the period between 1 April 1996 and 31 March 2005. The postnatal clinical features, final diagnoses and outcomes were examined to retrospectively compare these cases with biometric parameters obtained from routine antepartum ultrasound examinations. RESULTS: Final diagnoses included skeletal dysplasia (16), small-for-dates (SFD) infant without any abnormalities (seven), chromosomal abnormality (three), pyruvate dehydrogenase complex deficiency (one), Marden-Walker syndrome (one), and suspected Freeman-Sheldon syndrome (one). One turned out to be a healthy infant. All cases were divided on the basis of the final diagnoses into three groups: skeletal dysplasia (16 fetuses), SFD and healthy infant (eight fetuses) and others (six fetuses). The ratios of femur length (FL) to mean FL at a given gestational age (%FL) and of FL to biparietal diameter (FL/BPD) were significantly lower in the skeletal dysplasia group than those in the other groups. Moreover, in the skeletal dysplasia group, when the lethal cases were excluded, the ratio of FL to fetal trunk cross area (FL/FTA) was significantly lower than that in the other groups. CONCLUSIONS: FL/FTA appears to be a useful parameter to help differentiate fetuses with non-lethal skeletal dysplasia from anatomically normal fetuses either with constitutionally short limbs or with intrauterine growth restriction (IUGR).     

Target cells for an immunosuppressive cytokine, glycosylation-inhibiting factor.

Receptors for bioactive glycosylation-inhibiting factor (GIF) were demonstrated using a bioactive mutant of recombinant human (rh) GIF, which is comparable to the suppressor T (Ts) cell-derived bioactive GIF in its affinity for the receptors on helper T (Th) hybridoma cells. Both naive T and B cells in normal mouse spleen lacked GIF receptors. However, presentation of specific antigen to naive T cells resulted in the expression of the receptors on activated T cells. Furthermore, activation of small resting B cells with F(ab')2 fragments of anti-mouse IgM plus IL-4, lipopolysaccharide (LPS) plus IL-4 or LPS plus dextran sulfate induced the expression of the receptors within 48 h of B cell stimulation. It was also found that NK T cells freshly isolated from mouse spleen, but not conventional NK cells, expressed receptors for GIF. CD4(+) and CD4(-) subpopulations of NK T cells showed a similar binding capability. Mature dendritic cells derived from bone marrow did not bear the receptors. The dissociation constant (Kd) of the interaction between the bioactive rhGIF mutant and the high-affinity receptors was 10-100 pM, whereas inactive wild-type rhGIF failed to bind to the receptors. A bioactive derivative of rhGIF suppressed both IgG1 and IgE synthesis by purified B cells activated by LPS and IL-4, indicating that the binding of bioactive GIF to its receptors on activated B cells results in suppression of their differentiation.     

Migrainous infarction in an adult: evaluation with serial diffusion-weighted images and cerebral blood flow studies

We report the case of a 64-year-old man with migrainous infarction, giving special attention to chronological changes in neuroimaging findings. Five days after the onset, diffusion-weighted imaging showed slightly high intensity, and the apparent diffusion coefficient map showed increased diffusion in the right occipital lobe, which indicated vasogenic edema. Perfusion magnetic resonance imaging (MRI) and MR angiography demonstrated hyperperfusion of the ipsilateral hemisphere. Follow-up MRI showed irreversible brain damage. These images may reflect chronological changes in cerebral edema due to prolonged hyperperfusion with migraine.     

Discrimination of a nerve fiber that is the origin of a cauda equina tumor using acetylcholinesterase staining

Spinal nerve sheath tumors are well known to typically originate from dorsal sensory nerve roots. However, it is difficult to anatomically identify the origin in the case of cauda equina tumors. In this study, we aimed to ascertain whether a cauda equina nerve root removed with a nerve sheath tumor was a motor nerve using acetylcholinesterase (AchE) staining. Nerve rootlet sections removed with tumors were stained for AchE using the AchE Rapid Staining Kit. Additionally, we performed intraoperative motor‐evoked potential (MEP) monitoring following either transcranial electrical stimulation (TES) or electrical stimulation of nerve rootlets. The muscular strength of the lower extremities was assessed bilaterally before and after surgery using manual muscle testing. An AchE‐positive motor nerve rootlet that was the origin of a cauda equina tumor was observed in one of the 12 patients. In this patient, a MEP in the right quadriceps evoked by electrical stimulation of this rootlet was detected. TES‐MEP showed a 30% decrease in the amplitude in the right quadriceps evoked after tumor resection with this nerve rootlet. However, the motor strength in both lower extremities did not change after surgery. AchE staining and intraoperative MEP monitoring could detect the motor nerve rootlet that was the origin of a cauda equina tumor. Nerve sheath tumors originating from the motor nerve might be rare even in cauda equina.     

Differentiation between Treatment-Induced Necrosis and Recurrent Tumors in Patients with Metastatic Brain Tumors: Comparison among 11C-Methionine-PET,FDG-PET,MR Permeability Imaging,and MRI-ADC—Preliminary Results

BACKGROUND AND PURPOSE:In patients with metastatic brain tumors after gamma knife radiosurgery, the superiority of PET using ¹¹C-methionine for differentiating radiation necrosis and recurrent tumors has been accepted. To evaluate the feasibility of MR permeability imaging, it was compared with PET using ¹¹C-methionine, FDG-PET, and DWI for differentiating radiation necrosis from recurrent tumors.MATERIALS AND METHODS:The study analyzed 18 lesions from 15 patients with metastatic brain tumors who underwent gamma knife radiosurgery. Ten lesions were identified as recurrent tumors by an operation. In MR permeability imaging, the transfer constant between intra- and extravascular extracellular spaces (/minute), extravascular extracellular space, the transfer constant from the extravascular extracellular space to plasma (/minute), the initial area under the signal intensity–time curve, contrast-enhancement ratio, bolus arrival time (seconds), maximum slope of increase (millimole/second), and fractional plasma volume were calculated. ADC was also acquired. On both PET using ¹¹C-methionine and FDG-PET, the ratio of the maximum standard uptake value of the lesion divided by the maximum standard uptake value of the symmetric site in the contralateral cerebral hemisphere was measured (¹¹C-methionine ratio and FDG ratio, respectively). The receiver operating characteristic curve was used for analysis.RESULTS:The area under the receiver operating characteristic curve for differentiating radiation necrosis from recurrent tumors was the best for the ¹¹C-methionine ratio (0.90) followed by the contrast-enhancement ratio (0.81), maximum slope of increase (millimole/second) (0.80), the initial area under the signal intensity–time curve (0.78), fractional plasma volume (0.76), bolus arrival time (seconds) (0.76), the transfer constant between intra- and extravascular extracellular spaces (/minute) (0.74), extravascular extracellular space (0.68), minimum ADC (0.60), the transfer constant from the extravascular extracellular space to plasma (/minute) (0.55), and the FDG-ratio (0.53). A significant difference in the ¹¹C-methionine ratio (P < .01), contrast-enhancement ratio (P < .01), maximum slope of increase (millimole/second) (P < .05), and the initial area under the signal intensity–time curve (P < .05) was evident between radiation necrosis and recurrent tumor.CONCLUSIONS:The present study suggests that PET using ¹¹C-methionine may be superior to MR permeability imaging, ADC, and FDG-PET for differentiating radiation necrosis from recurrent tumors after gamma knife radiosurgery for metastatic brain tumors.

Stereotactic radiosurgery such as gamma knife radiosurgery (GK) and CyberKnife (Accuray, Sunnyvale, California) is an effective method for treating intracranial neoplasms.^1,2 For metastatic tumors of the brain, stereotactic radiosurgery has generally been the main tool used in therapeutic regimens.^3,4 Although stereotactic radiosurgery is an effective treatment method, it has a risk of radiation necrosis. Radiation necrosis after stereotactic radiosurgery for metastatic tumors of the brain is more common than previously reported.^5,6 It generally occurs 3–12 months after therapy⁷ and often resembles recurrent tumors on conventional imaging techniques, such as MR imaging,^8–11 CT,¹² and SPECT.¹³ Differentiating radiation necrosis and recurrent tumor is extremely important because of the different treatment implications. Histologic examination from a biopsy or resection may aid in differentiating these 2 events. However, a noninvasive method is needed for diagnosing whether a contrast-enhanced lesion with surrounding edema on conventional MR imaging is radiation necrosis or a recurrent tumor.Advanced MR imaging techniques including MR spectroscopy,¹⁴ DWI,¹⁵ and DTI¹⁶ have been used for differentiation of radiation necrosis and recurrent tumors. The CTP technique has also been reported as promising in this field.¹⁷ CTP has the advantage of using widely available CT scanners, though x-ray exposure and administration of ionizing contrast material limit the clinical use. In radionuclide studies, SPECT with ²⁰¹TI-chloride,¹⁸ technetium Tc99m-sestamibi,^{19 123}I-alfa-methyl-L-tyrosine,²⁰O-(2-[¹⁸F]-fluoroethyl)-L-tyrosine (FET-PET),^21,22 6-[¹⁸F]-fluoro-L-dopa (FDOPA),²³ and FDG-PET^24–26 have been reported to differentiate between radiation necrosis and recurrent tumors. Compared with those studies, the superiority of PET with ¹¹C-methionine (MET) for differentiating radiation necrosis and recurrent tumors has been accepted because of the high sensitivity and specificity.^27–31 However, MET-PET is not widely available. Dynamic contrast-enhanced MR imaging with a contrast agent has been used to characterize brain tumors^32,33 and stroke.³⁴MR permeability imaging with dynamic contrast-enhanced–MR imaging based on the Tofts model³⁵ has recently been developed and used for evaluating cerebrovascular diseases,³⁶ brain tumors,^37–39 nasopharyngeal carcinomas,^40,41 rectal carcinomas,⁴² and prostate carcinomas.⁴³ The endothelial permeability of vessels in brain tumors can be quantitatively acquired with MR permeability imaging. The vascular microenvironment in tumors can be measured by parameters such as influx transfer constant, reverse transfer constant, and the extravascular extracellular space.⁴⁴ These parameters may reflect tissue characteristics including vascular density, a damaged blood-brain barrier, vascularity, and neoangiogenesis.⁴⁴ If the feasibility of MR permeability imaging for differentiating radiation necrosis and recurrent tumors could be demonstrated, this technique may contribute to the management of patients after stereotactic radiosurgery and conventional radiation therapy because MR permeability imaging is widely available. To evaluate the feasibility of MR permeability imaging in the present study, we compared it with MET-PET, FDG-PET, and DWI for differentiating radiation necrosis from recurrent tumor after GK in patients with metastatic brain tumors.     

Two cases of colorectal cancer patients with poor performance status who had unresectable liver metastasis effectively treated by cetuximab

We reported two cases of colorectal cancer patients with EGFR-positive unresectable synchronous liver metastasis effectively treated by cetuximab after the progression of the prior chemotherapy. Case 1: A 49-year-old female with unresectable synchronous liver metastasis from colon cancer received cetuximab monotherapy as fifth-line therapy. Then, abdominal CT showed shrinkage of the liver metastasis (PR) and the performance status was improved from 3 to 0 as upper abdominal pain reduced. Case 2: A 67-year-old female with unresectable liver metastasis from colon cancer received cetuximab with CPT-11 combined therapy as fourth-line therapy. After that, liver metastasis also decreased (PR), and upper abdominal pain and PS were improved from 2 to 0. These two cases of KRAS status on cancer tissue also showed wild-type, and in these cases cetuximab proved effective.     

Analogy between sphere forming ability and stemness of human hepatoma cells

Increasing evidence suggests that cancers contain a small subset of cancer-initiating cells, so-called cancer stem cells (CSCs) that are capable of regenerating a tumor after chemoradiation therapy. Sphere forming ability is known to be one of properties of CSCs, but the significance remains unclear. The present study focused on sphere formation of human hepatoma cells in three-dimensional culture in order to evaluate the analogy between sphere forming ability and stemness of cancer cells in vitro. Under three-dimensional culture condition, HepG2, Hep3B and PLC/PRF/5 cells demonstrated the sphere formation while SK-Hep1 and Huh-7 cells did not. The population of G0/G1 phase increased in the spheres compared with the monolayer (67 vs. 38%). In spite of no significant difference in stem cell surface markers (CD44, CD90, CD133, EpCAM and ABCG2), remarkable up-regulation of p27 CDK inhibitor was observed in sphere forming cells. Immunofluorescence analysis revealed the nuclear expression of p27 in the whole of the sphere, but weak expression of p21 only at the peripheral area. The spheres acquired chemoresistance to cisplatin compared with the monolayers (58.9 vs. 16.2 μM in IC50). This model was useful for assessment of the role of cell-cycle quiescence in the stemness and chemoresistance of cancer cells.