The map kinase ERK regulates renal activity of cyclin-dependent kinase 2 in experimental glomerulonephritis.

BACKGROUND: In vitro, the extracellular signal-regulated kinase (ERK) is an intracellular convergence point of multiple stimuli, which affect the cell cycle. However, the role of ERK in cell cycle regulation in vivo is unknown. METHODS: To address this issue, ERK activity was blocked both in vitro in mesangial cells (MC) and in vivo in experimental glomerulonephritis (GN) by a pharmacological inhibitor (U0126) of the ERK-activating kinase. RESULTS: In stimulated MC, inhibition of ERK reduced cyclin-dependent kinase 2 (CDK2) phosphorylation, CDK2 activity and cyclin E/A expression, whereas downregulation of CDK inhibitor p27(Kip1) expression was inhibited. In vivo, U0126 was given to rats in the acute phase of anti-Thy 1.1 GN. We previously showed that glomerular cell proliferation was reduced by 67% upon treatment with the inhibitor compared to nephritic controls. Now, we detected a significant increase in renal CDK2-activity/phosphorylation in the nephritic controls, that was significantly and dose-dependently reduced by ERK inhibition. CDK2 activation was accompanied by an increase in renal expression of cyclins E/A and the enhanced binding of these cyclins to CDK2 in the nephritic controls. These changes were blunted by U0126 treatment. Finally, we noted an increased expression and CDK2-binding of p27(KIP1) protein in the nephritic controls which was decreased in U0126 treated rats. CONCLUSIONS: Our observations provide the first evidence that ERK is an intracellular regulator of renal CDK2 activity in vivo in a glomerulonephritis model.     

Sentinel node sampling limits lymphadenectomy in stage I non-small cell lung cancer.

OBJECTIVE: It is controversial whether a systematic mediastinal lymph node dissection (MLND) needs to be performed in all patients with stage I lung cancer. The present study was done to examine the new sentinel lymph nodes hypothesis based on the lobe of the primary tumor. METHODS: In our first study, the lymph node (LN) metastases were assessed in 291 stage I non-small cell lung cancer (NSCLC) patients who had a major lung resection with a systematic mediastinal lymph node dissection. We evaluated the validity of using our new sentinel lymph nodes method based on the lobe of the primary tumor as follows: the pretracheal (#3), tracheobronchial (#4), and hilar nodes (#10) for right upper lobe tumors; #4, subcarinal (#7), and #10 for middle lobe tumors; the subaortic (#5), paraaortic (#6), and #10 for left upper lobe tumors; and the #7, #10, and interlobar nodes (#11) for tumors in either lower lobes. In the second study, we performed a lobectomy with new sentinel node sampling in 64 patients with preoperative complications. If all of the sampling nodes showed no metastases on frozen section diagnosis, systematic node dissections were not performed. RESULTS: Six of 291 patients in the first study had skip metastases that did not involve the new sentinel nodes; 5 of the 6 patients had macroscopic pleural invasion. Thus, we defined pleural invasion as an exclusion criterion for the second study. In the second study, the median follow-up time was 39 months. Metastatic lymph nodes were detected in 11 of 64 patients. Fifty-three patients (83%) had no metastasis in the sampled nodes, and, therefore, a mediastinal lymph node dissection was not done. The morbidity rate in the sampling group was 36%, and there was no mortality. In the sampling group, local recurrences were observed in two patients, distant metastases in eight, and carcinomatous pleuritis in one; the overall 5-year survival rate was 82%. CONCLUSIONS: We found that it is possible to perform a less invasive lymphadenectomy for patients with stage I lung cancer using intra-operative sampling of new sentinel lymph nodes.     

Two cases of atraumatic iliopsoas hematoma

We report two cases of atraumatic iliopsoas hematoma. First patient was a 76-year-old man admitted to our hospital from appetite loss. Blood transfusion did not improve his anemia. Five days after admission, suddenly he went into shock. CT scan revealed ileopsoas hematoma. He died from hemorrhagic shock in spite of conservative therapy. Second patient was a 70-year-old man admitted because of acute heart failure. Continuous hemodiafiltration was required to relieve anuria. The next day, he developed left leg and hip pain. CT scan revealed ileopsoas hematoma and he received CT guided aspiration drainage for decompression, but almost 7 days were needed to achieve successful pain control. In a case of iliopsoas hematoma, early diagnosis and adequate choise of therapy are necessary to improve prognosis of patients.     

Clinical Course and Autopsy Findings of a Patient with Clival Chordoma Who Underwent Multiple Surgeries and Radiation during a 10-Year Period.

The management of clival chordoma remains problematic. We present the case of a 48-year-old woman with clival chordoma who underwent multiple surgeries and radiation therapy, including gamma knife stereotactic radiosurgery (GK-SRS), during a 10-year clinical course. The tumor was initially removed by gross total resection via the trans-sphenoidal approach, followed by external linac radiation therapy. The tumor recurred at the clivus 5 years after the initial operation. After repeated trans-sphenoidal removal of recurrent tumors, she twice underwent GK-SRS for a tumor remnant adjacent to the brainstem. Although this part of the tumor was controlled by GK-SRS, there was further tumor extension toward the sphenoid and maxillary sinuses. Ultimately, lower cranial nerve dysfunction developed due to tumor extension into the lower part of the clivus and the patient died of respiratory failure. Autopsy revealed the tumor to extend from the lower clivus to the bilateral middle fossae. The lower part of the tumor extended to the nasal cavity and to the posterior wall of the pharynx, resulting in compression of the upper pharyngeal region. The tumor around the jugular foramen compressed the lower cranial nerves bilaterally. Tumor cells did not, however, invade the intradural space microscopically. Although chordoma is not biologically malignant, this tumor can show massive extension with destruction of bony structures and extracranial invasion of connective tissues. Therefore, the optimal treatment strategy is to remove the tumor mass as extensively as possible, including normal bony structures and connective tissues surrounding the tumor, using skull base surgical techniques.     

Therapeutic potential of transgenic mesenchymal stem cells engineered to mediate anti–high mobility group box 1 activity: targeting of colon cancer

Background

Mesenchymal stem cells (MSCs) are being developed as a new clinically relevant stem cell type to be recruited into and to repair injured tissue. A number of studies have focused on the therapeutic potential of MSCs by virtue of their immunomodulatory properties. Systemically administered MSCs can also migrate to sites of malignancies. Because of this latter phenomenon, we transfected human MSCs to secrete anti–high mobility group box (HMGB) 1 proteins. They were then injected into mice bearing human colon cancer to evaluate their efficacy as an antineoplastic agent.

Materials and methods

The ABOX gene was used in this model, which encodes part of the HMGB1 protein and acts as an HMGB1 antagonist. It was cotransduced by electroporation with a FLAG-tag to visualize the secreted ABOX protein, levels of which in supernatants from cultured transfected MSCs were quantified by immunofluorescence imaging using an anti-FLAG antibody. Antiangiogenic effects were evaluated in vitro using a novel optical assay device for the quantitative measurement of cellular chemotaxis assessing the velocity and direction of endothelial cell movement stimulated by supernatant from tumor cells. We found that ABOX proteins released from transfected MSCs suppressed migration in this assay. Finally, MSCs were injected subcutaneously into Nonobese diabetic/severe combined immunodeficiency mice bearing human colon cancer from a cell line, which secreted large amounts of HMGB1. Ten days after MSC injection, mice were sacrificed and tumors evaluated by immunohistochemistry.

Results

From 12 ho through 7 d after gene transfection, ABOX proteins secreted from MSCs could be detected by immunofluorescence and enzyme-linked immunosorbent assay. Quantitative measurement of cellular chemotaxis demonstrated that ABOX proteins secreted from transfected MSCs decreased the velocity and interfered with the direction of movement of vascular endothelial cells. Moreover, in an in vivo human colon cancer xenograft model, injection of anti-HMGB1–transfected MSCs resulted in a decreased tumor volume due to the antiangiogenic properties of the secreted ABOX proteins.

Conclusions

MSC modified to secrete HMGB1 antagonist proteins have therapeutic antineoplastic potential. These findings may contribute to future novel targeting strategies using autologous bone marrow–derived cells as gene delivery vectors.     

Low-Volume Walking Program Improves Cardiovascular-Related Health in Older Adults

Although numerous sources of evidence show that regular physical activity is beneficial to health, most individuals do not engage in a sufficient amount of physical activity to meet the guidelines set out by expert panels. In addition, the minimum amount of physical activity associated with reduced cardiovascular disease risk markers is not clear in older adults. The purpose of this study was to determine the effects of a 12-week walking program involving an exercise volume below the current minimum physical activity recommendation on cardiovascular disease risk markers in older adults. The participants were recruited from the following two groups separately: a walking group (n = 14) and a control group (n = 14). In the walking group, participants walked 30 to 60 minutes per session on 2 days per week for 12 weeks (average walking time, 49.4 ± 8.8 min/session). Plasma oxidised low-density lipoprotein concentrations tended to be lower than baseline values in the walking group after 12 weeks (paired t-test, p = 0.127). The ratio of oxidised low-density lipoprotein to high-density lipoprotein cholesterol was significantly lower than the baseline ratio in the walking group after 12 weeks (paired t-test, p = 0.035). Resting systolic blood pressure and diastolic blood pressure were significantly lower than baseline values in the walking group after 12 weeks (paired t-tests, p = 0.002, p < 0.0005, respectively). Our findings demonstrate that a 12-week walking program comprising a low volume of physical activity confers a benefit to cardiovascular-related health in older adults.

Key Points

• It is important to consider baseline physical activity levels when evaluating physical activity program.
• Being physically active is important to reduce the potential risk marker of cardiovascular disease in older adults.
• These data imply that a small volume of 12-week walking program confers a benefit to cardiovascular-related health in older adults.

Key words: blood pressure, exercise, lipid metabolism, older adults, oxidised low-density lipoprotein     

Screw perforation features in 129 consecutive patients performed computer-guided cervical pedicle screw insertion

Study design

A cross-sectional study of the data retrospectively collected by chart review.

Objectives

This study aimed to clarify screw perforation features in 129 consecutive patients treated with computer-assisted cervical pedicle screw (CPS) insertion and to determine important considerations for computer-assisted CPS insertion.

Summary of background data

CPS fixation has been criticized for the potential risk of serious injury to neurovascular structures. To avoid such serious risks, computed tomography (CT)-based navigation has been used during CPS insertion, but screw perforation can occur even with the use of a navigation system.

Methods

The records of 129 consecutive patients who underwent cervical (C2–C7) pedicle screw insertion using a CT-based navigation system from September 1997 to August 2013 were reviewed. Postoperative CT images were used to evaluate the accuracy of screw placement. The screw insertion status was classified as grade 1 (no perforation), indicating that the screw was accurately inserted in pedicle; grade 2 (minor perforation), indicating perforation of less than 50 % of the screw diameter; and grade 3 (major perforation), indicating perforation of 50 % or more of the screw diameter. We analyzed the direction and rate of screw perforation according to the vertebral level.

Results

The rate of grade 3 pedicle screw perforations was 6.7 % (39/579), whereas the combined rate of grades 2 and 3 perforations was 20.0 % (116/579). No clinically significant complications, such as vertebral artery injury, spinal cord injury, or nerve root injury, were caused by the screw perforations. Of the screws showing grade 3 perforation, 30.8 % screws were medially perforated and 69.2 % screws were laterally perforated. Of the screws showing grades 2 and 3 perforation, 21.6 % screws were medially perforated and 78.4 % screws were laterally perforated. Furthermore, we evaluated screw perforation rates according to the vertebral level. Grade 3 pedicle screw perforation occurred in 6.1 % of C2 screws; 7.5 % of C3 screws; 13.0 % of C4 screws; 6.5 % of C5 screws; 3.2 % of C6 screws; and 4.0 % of C7 screws. Grades 2 and 3 pedicle screw perforations occurred in 12.1 % of C2 screws, 22.6 % of C3 screws, 31.5 % of C4 screws, 22.2 % of C5 screws, 14.4 % of C6 screws, and 12.1 % of C7 screws. C3–5 screw perforation rate was significantly higher than C6–7 (p = 0.0024).

Conclusions

Careful insertion of pedicle screws is necessary, especially at C3 to C5, even when using a CT-based navigation system. Pedicle screws tend to be laterally perforated.     

Cloning and characterization of three PHEX homologues in <Emphasis Type="Italic">Drosophila</Emphasis>

Inactivating mutations and/or deletions of PHEX (Phosphate-regulating gene with Homologies to Endopeptidase on the X chromosome) are responsible for X-linked hypophosphatemic rickets in humans. In the present study, three Drosophila PHEX homologues (dPHEX-1, -2, -3) were isolated by the screening of a Drosophila cDNA library and expressed sequence tag (EST) database. The structural region involving motif II: ⁴⁵⁶WMXXXTKXXAXXK⁴⁶⁸ (numbered according to human PHEX), motif VI: ⁶⁰²WW⁶⁰³, and motif VIII: ⁷⁴⁶CXLW⁷⁴⁹ was conserved in the dPHEX family. Zinc-coordinating motifs (HEFTH and GENIADNGG) were also conserved in the dPHEX family. All three dPHEX genes were expressed during all stages of Drosophila development. The expression of dPHEX-1 was suppressed by dietary phosphate deprivation, but the expression of dPHEX-2 and that of dPHEX-3 were not affected. In-situ hybridization showed a ubiquitous distribution of dPHEX-1 and dPHEX-2, while dPHEX-3 was highly expressed in the larval brain. In an analysis of subcellular localization, dPHEX-1 was localized to intracellular organelles and dPHEX-3 was localized predominately in the plasma membrane of Drosophila embryonic S2 cells. Homozygosity of a dPHEX-1 mutation, a transposon insertion in the dPHEX-1 promoter region, was completely lethal at an early stage of embryonic development. The present study indicates that three homologues are likely involved in the phosphate homeostasis of Drosophila.     

Diagnosis of deep vein thrombosis using multi-detector helical CT

PURPOSE: To evaluate the usefulness of multi-detector helical CT (MDHCT) with contrast medium in the diagnosis of deep vein thrombosis (DVT). Materials and Methods: The bilateral veins of the dorsal pedis in 45 patients (12 men, 33 women; average age, 64 years) under clinical suspicion of DVT were first punctured using 22-G needles. Then CT scanning from the level of the foot to the inferior vena cava was started 20 sec after the initial injection of 200 mL of dilute contrast medium (50 mL nonionic iodinated contrast medium of 300 mgI/mL and 150 mL saline) at a rate of 5 mL/sec. RESULTS: Two patients were excluded because of unsuccessful venous puncture. The average scanning time in 43 patients was 38.5 +/- 7.9 seconds. Images of veins from the foot to the inferior vena cava were clearly demonstrated in each case. MDHCT showed DVT in 32 cases and patent deep vein in 11 cases. Simultaneous venography of the lower extremity in 18 patients clearly visualized DVT at the same level detected by contrast MDHCT. CONCLUSION: MDHCT for the diangosis of DVT has the advantages of wider scanning range, shorter scanning time, and finer Z-axis resolution than the other diagnostic modalities.     

The retention indices of 201Tl-SPECT in brain tumors

OBJECTIVE: The aim of this study was to assess the utility of 201Tl SPECT in the differential diagnosis of intracranial tumors and to determine the relationship between 201Tl uptake and histological types. METHODS: Thirty-eight patients (19 males and 19 females) with thirty-eight brain tumors were evaluated with 201Tl-SPECT. The early and delayed 201Tl uptake ratio was calculated, and the retention index (RI) was applied as follows; RI = delayed uptake ratio/early uptake ratio. RESULTS: The RI of malignant tumors was higher (0.72 +/- 0.18) than that of benign tumors (0.50 +/- 0.16) and the difference was statistically significant (p = 0.00045). The difference between high-grade glioma (0.80 +/- 0.15) and metastatic tumors (0.64 +/- 0.19) was statistically significant (p = 0.039). CONCLUSION: 201Tl-SPECT may add useful biochemical information and could differentiate malignant brain tumors from benign lesions, but the RI of metastatic tumors varied depending on the organs with the primary lesion and histological types.