Sleep Disturbance in Children with Autism Spectrum Disorder: A Cross Sectional Study.

Sleep problems are commonly seen in children with Autism Spectrum Disorder (ASD). According to previous research, sensory problems and anxiety may be related to the development and maintenance of sleep problems in children with ASD. To determine pattern and severity sleep disturbance in children with autism spectrum disorder. This descriptive cross sectional study was done in Institute of Paediatric Neuro-disorder and Autism (IPNA) and Department of Paediatric Neurology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh from October 2017 to September 2018. A total of 59 children aged 3 to 15 year who were diagnosed as ASD according to the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) were enrolled in this study. Sleep disturbances were recorded in the standard questionnaire. The mean age was 49.78±20.69 months and male to female ratio was almost 4:1. More than two third (69.2%) patients had sleep awakening in mild, 7(36.8%) in moderate autism spectrum disorder. The difference was statistically significant (p<0.05) among three groups. More than half (56.4%) patients had sleep starting 12 am to 2 am in mild, 3(15.8%) in moderate autism spectrum disorder. The difference was statistically significant (p<0.05) among three groups. Sleep awakening and sleep starting time were significantly associated with autism spectrum disorder.

Relaparotomy Following Caesarean Section: A Cross Sectional Study.

Delivery by caesarean section (CS) is the most common obstetric procedure in daily practice and rate is increasing worldwide. In spite of huge appeal for this operation, there is significant rate of short & long term complications. Relaparotomy after caesarean section in early post-operative period is one of the rarest short term complications but the fatality rate is very high. The objective of this study was to find out indications, managements, risk factors and outcomes of patients undergoing relaparotomy after CS. Finally to improve the quality of care for preventing this dreadful complication and to reduce the maternal mortality and morbidity. Purposive sampling was done, all consecutive patients who underwent relaparotomy within 6 weeks of CS in Rajshahi Medical College Hospital (RMCH) during study period from January 2015 to December 2015 has been included for this study. This was a cross-sectional and observational study in a tertiary referral and teaching hospital RMCH. During study period total admitted obstetric patients at RMCH were 12688. There were 9802 deliveries where 53.89% (n=5282) had vaginal delivery and 46.11% (n=4520) underwent caesarean sections. Among these 46.11% (n=4520) CS, relaparotomy was needed 0.18% (n=8) cases. Total relaparotomy was done in 0.39% (n=50) cases out of 12688 obstetric patients. Out of 50 cases 42 had caesarean delivery in other hospitals and clinics outside RMCH. The indications of relaparotomy were secondary post-partum hemorrhage (PPH) 28% (n=14), primary PPH 12% (n=6), haemoperitoneum 22% (n=11), pyoperitoneum 18% (n=9), subrectal hematoma 16% (n=8) and burst abdomen 4% (n=2). Main surgeries performed were subtotal hysterectomy in 44% (n=22) cases, total abdominal hysterectomy in 10% (n=5) cases, re-suturing of uterine incision 8% (n=4), drainage of pus & peritoneal toileting 8% (n=4), ligation of bleeding vessels 6% (n=3), drainage of subrectalhaematoma in 16% (n=8) cases, repair of intestinal injury 4% (n=2) and repair of anterior abdominal wall in 4% (n=2) cases. Case fatality of relaparotomy was high 18% (n=9), majority were preventable. Identification of risk factors, adequate attention, expert decision, prompt intervention & proper management will improve the outcome.

Induction by Misoprostol In Case of Intra Uterine Fetal Death: A Cross Sectional Study.

The objective of this study was to find out the effectiveness and safety of vaginal misoprostol in delivering the dead fetus in cases of intrauterine fetal death (IUFD). This cross sectional study was undertaken among all consecutive patients admitted at the Department of Obstetrics and Gynecology, Dhaka Medical College Hospital, Dhaka, Bangladesh, from October 2012 to September 2013, were included for this study. Vaginal misoprostol was applied in 50 cases that were admitted and diagnosed as a case of IUFD. After taking informed written consent from patients 50µgm of tablet misoprostol was introduced per vaginally into the posterior fornix. Doses were repeated in every 4 hours up to effective contraction to a maximum 6 doses. Follow up was done in hourly interval. Those who did not respond, decision for other methods like oxytocin infusion or LUCS were done. Study showed 60% (n=30) patients belonged to age group of 18-24 years. Among study population 44% (n=22) of the patients belonged to 22-32 weeks of gestation, 30% (n=15) were 33-37 weeks and 26% (n=13) were 38-42 weeks. The causes of intrauterine death were unidentified among 60% (n=30) of cases. Regarding antenatal check up 54% (n=27) had no checkup, 26% (n=13) were irregular and 20% (n=10) were regular. 60% (n=30) of the patients had less Bishop Score <3. Ninety six percent (96%) (n=48) patients responded with vaginal misoprostol and only 4% (n=2) were non responder group. Most 84% (n=42) of the cases needed 2-3 doses, only 8% (n=4) cases needed single dose and 8% (n=4) cases needed 4 to 6 doses. The induction delivery interval varied from 6 to 23 hours and maximum 52% (n=26) of the patients delivered within 12 hours. Adverse effects such as uterine hyper stimulation, tachysystole were not detected. The major complications observed like postpartum hemorrhage 4% (n=2), reduced platelet count 2% (n=1), and chorioamnionitis 4% (n=2). However minor complications like nausea, shivering and mild fever also observed in few cases. Study showed successful induction of labor by misoprostol in a shorter time with significantly less side effects.

Childhood Neuro-developmental Disorder (Rett Syndrome): A Case Report.

Rett syndrome is a disorder of early brain development which is clinically characterized by arrested neuro-development. We found one such 5.5 years old girl whose physical and mental development was normal up to 17 months of age, followed by regression. She had lost her already acquired purposeful hand movements, appearance of stereotyped hand movements, along with development of epilepsy. To our knowledge such case is being reported for the first time from Bangladesh. The purpose of this case report is to increase awareness of this syndrome among physicians specially paediatricians, thereby aiding early diagnosis and treatment.

Neuro-Imaging Changes in Cerebral Palsy: A Cross Sectional Study.

Cerebral palsy (CP) is a non- progressive disorder of movement and posture due to a lesion of the developing brain. It is the commonest physical disability in childhood that affects function and development. Neuro imaging is currently recommended as a standard evaluation in children with cerebral palsy. This hospital based cross-sectional study was conducted in Paediatric Neurology out-patient department of Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh from July 2015 to December 2015 to see the frequency and pattern of neuro-imaging findings in children with cerebral palsy. A total of 130 cases those who were attended and diagnosed as cerebral palsy based on history, clinical examination and neuro developmental assessment included in this study. All patients were sent to radiology & imaging department of same hospital for CT scan of brain. Among total 130 cerebral palsy patients male were more affected than female (88 boys and 42 girls) with male to female ratio 2.09: 1. Their ages ranged between 6-72 months with a mean age 25.6 months. The commonest age group was 6-24 months (46.9%). Common mode of delivery was normal vaginal delivery (62.3%) & Perinatal asphyxia (PNA) occurred in 66.9% cases. The commonest type of cerebral palsy was spastic form. Among them most cases were quadriplegic type, 64 cases (53.3%). Other cases were hemiplegic 27(20.7%) diplegic 13(10.0%). Total 84.7% had documented cerebral neuroimaging abnormalities; among them, diffuse cortical atrophy (46.9%), encephalomalacic change (19.9%), malformation (6.1%), and calcification (5.3%). CT scan was normal in 15.3% cases of cerebral palsy. The commonest co morbidity was speech delay (50%). Most of the patient with CP had abnormal CT scan finding though some patient had normal CT scan. Diffuse cerebral atrophy and encephalomalacic changes constitute frequent CT neuroimaging findings and commonly found in quadriplegic type of cerebral pulsy patients. Though diagnosis of cerebral palsy is essentially clinical, neuro imaging improves the understanding of the neuro-anatomical basis for function in CP. Etiology, type of CP and extent of motor impairments can easily be identified by the neuro imaging.

Clinical and Neuro-radiological Presentation of Tuberous Sclerosis Complex in Tertiary Care Centre.

Tuberous sclerosis complex (TSC) is a common neurocutaneous disorder characterized by hamartomatous changes in the lungs, brain, kidneys, skin, heart, and other organs. This retrospective study was done to see the clinical presentation and neuro imaging pattern of TSC in a tertiary care centre of Bangladesh and was conducted at Pediatric Neurology Unit of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from January 2013 to December 2013. Among total 10 patients male-female ratio was 3:2. Commonest age group was 1-5 year in 50%. Common cutaneous abnormality was facial angiofibroma in 100%. Commonest neurological presentation was epilepsy in100%, ASD in 20% patient. Common renal manifestation was multicystic kidney disease (20%). CT/MRI findings were periventricular calcifications in 70% patients. In this study facial angiofibroma is the most common skin manifestation and epilepsy is the common neurological presentationandperiventricular calcification is the most common neuro-radiological findings in tuberous sclerosis complex.

Non-migratory tumorigenic intrinsic cancer stem cells ensure breast cancer metastasis by generation of CXCR4(+) migrating cancer stem cells.

Although the role of metastatic cancer stem cells (mCSCs) in tumor progression has been well documented, our study reveals a hitherto unidentified role of tumorigenic intrinsic CSCs (iCSCs) in breast cancer metastasis. We show that unlike highly migratory mCSCs residing in the breast tumor disseminating/peripheral regions, iCSCs populate the inner mass of the tumor and are non-migratory. However iCSCs, via paracrine signaling, induce conversion of non-stem cancer cells to CSCs that (i) are identical to the previously reported mCSCs, and (ii) in contrast to iCSCs, express chemokine receptor, chemokine (C-X-C motif) receptor 4 (CXCR4), which is crucial for their metastatic potential. These mCSCs also demonstrate high in vivo tumorigenicity. Physical non-participation of iCSCs in metastasis is further validated in vivo, where only mCSCs are found to exist in the metastatic sites, lymph nodes and bone marrow, whereas the primary tumor retains both iCSCs and mCSCs. However, iCSCs ensure metastasis since their presence is crucial for deliverance of highly metastatic CXCR4(+) mCSCs to the migrating fraction of cells. Cumulatively, these results unveil a novel role of iCSCs in breast cancer metastasis as parental regulators of CXCR4(+) mCSCs, and highlight the therapeutic requisite of targeting iCSCs, but not CXCR4(+) mCSCs, to restrain breast cancer metastasis from the root by inhibiting the generation of mCSCs from iCSCs. Considering the pivotal role of iCSCs in tumor metastasis, the possibility of metastasis to be a 'stem cell phenomena' is suggested.

Bickerstaff's Brainstem Encephalitis - A Case Report.

A 11 years male child was presented with fever, headache, vertigo, repeated seizure for 8 days. He had also complains of hemiperesis of right side and impaired vision. On examination patient was drowsy and somnolent. Paralysis of circular muscles of eye ball, exaggerated deep reflexes, extensor planter reflexes and right sided ataxia were present. All routine investigations revealed normal findings except CSF showed slight pleocytosis. MRI of brain revealed hyper intensity of midbrain in left side. The diagnosis of Bickerstaff's brainstem encephalitis (BBE) was strongly suggested from the medical history, clinical examination, biochemical and radiological (MRI) findings of brain. The purpose of our report is to highlight this very rare disease to all.

Semaphorin 3A upregulates FOXO 3a-dependent MelCAM expression leading to attenuation of breast tumor growth and angiogenesis.

Semaphorin 3A (Sema 3A), a member of semaphorin family, serves as a guidance clue during embryonic development and is known as a candidate tumor suppressor that attenuates breast tumor progression by binding with its co-receptor, neuropilin-1 (NRP-1). However, the underlying mechanism by which Sema 3A suppresses breast tumor growth is still unexplored. In this study, we report that Sema 3A regulates phosphorylation and nuclear translocation of phosphatase and tensin homolog (PTEN) and FOXO 3a. Moreover, Sema 3A controls NRP-1-mediated PTEN-dependent FOXO 3a activation. Overexpression of PTEN and FOXO 3a enhances Sema 3A-induced attenuation of breast cancer cell migration. Chromatin immunoprecipitation and electrophoretic mobility shift assay data revealed that FOXO 3a regulates MelCAM at the transcriptional level. Furthermore, Sema 3A induces NRP-1-mediated MelCAM expression through PTEN and FOXO 3a. The data also showed that vascular endothelial growth factor-induced angiogenesis is inhibited by Sema 3A. Loss of or gain in function study revealed that Sema 3A modulates phosphorylation of PTEN and FOXO 3a and expression of MelCAM, leading to suppression of tumor growth and angiogenesis using in vivo mice model. Clinical specimen analysis revealed that reduced expression of Sema 3A and p-PTEN are correlated with enhanced breast cancer progression, further strengthening our in vitro and in vivo findings. Correlation of relapse-free survival of breast cancer patients (n=2878) with expression levels of Sema 3A, NRP-1, FOXO 3a and MelCAM were studied by Kaplan-Meier analysis. Statistical analysis revealed a close association between reduced expression of Sema 3A and MelCAM with that of poor patient's survival. Our study demonstrated a novel mechanism of regulation of tumor suppression by Sema 3A in coordination with a chain of tumor-suppressor genes, which in turn inhibits breast cancer cell migration, tumor growth and angiogenesis.

Biocompatible amphiphilic pentablock copolymeric nanoparticles for anti-cancer drug delivery.

A pentablock copolymer of Poly(Lactide-co-Glycolide) and Pluronic F68 was synthesized using ring-opening polymerization and characterized by NMR and FTIR for confirming the structure of the block copolymer. TG-DTA studies showed PLGA:Pluronic ratio to be 4:1. As the PLGA-PEO-PPO-PEO-PLGA Pentablock Copolymer (PPPC) prepared is amphiphilic, its Critical Vesicular Concentration, was measured, which was lower at 37 degrees C than at 25 degrees C, which could provide better stability to the system at physiological temperature. The nanoparticles of PPPC vary in topographyand range from 150 to 500 nm in size, according to the synthesis route used viz Emulsion Solvent Evaporation and simple dialysis. Pentablock copolymer nanoparticles were found to entrap about 84% of hydrophobic drug, docetaxel. Drug release profile of docetaxel showed about 50% release in first 2 hours at pH 4.6 and about 80% docetaxel was released at pH 7.4, at the end of 2 days. The PPPC nanoparticles was found to be biocompatible to L929 cell lines up to 1 mg/ml concentration. Preliminary in vitro cytotoxic effect of docetaxel loaded PPPC nanoparticles against four different cancer cell lines showed 50% inhibitory concentration of 6 nM in A431 (Squamous cell carcinoma), 250 nM in HeLa (Cervical carcinoma), 800 nM in PC3 (Prostate carcinoma) and 1 microM in KB (Epidermoid carcinoma) cells.

Hypoxia-driven osteopontin contributes to breast tumor growth through modulation of HIF1α-mediated VEGF-dependent angiogenesis.

Hypoxia is a salient feature of most solid tumors, and hypoxic adaptation of cancer cells has crucial implications in propagation of malignant clonal cell population. Osteopontin (OPN) has been identified as a hypoxia-responsive gene, but the mechanistic and regulatory role of OPN under hypoxia is less characterized. The present study identifies the existence of a positive inter-regulatory loop between hypoxia and OPN. We have shown that hypoxia induces OPN expression in breast cancer cells; however, the expression was found to be HIF1α independent. OPN enabled transcriptional upregulation of HIF1α expression both under normoxia and hypoxia, whereas stability of HIF1α protein in breast cancer cells remained unaffected. Moreover, we have shown that OPN induces integrin-linked kinase (ILK)/Akt-mediated nuclear factor (NF)-κB p65 activation leading to HIF1α-dependent vascular endothelial growth factor (VEGF) expression and angiogenesis in response to hypoxia. These in vitro data are biologically important as OPN expressing cells induce greater tumor growth and angiogenesis through enhanced expressions of proangiogenic molecules as compared with control. Immunohistochemical analysis of human breast cancer specimens revealed significant correlation between OPN and HIF1α but not HIF2α. Elevated expression of HIF1α and OPN was observed in pre-neoplastic and early stage infiltrating ductal carcinoma implicating the role of these proteins in neoplastic progression of breast cancer. Together, our results substantiate the prime role of OPN in cellular adaptation through ILK and NF-κB-mediated HIF1α-dependent VEGF expression in response to hypoxia that ultimately controls breast cancer progression and angiogenesis. Our study reinforces the fact that targeting OPN and its regulated signaling network hold important therapeutic implications.

Osteopontin signaling upregulates cyclooxygenase-2 expression in tumor-associated macrophages leading to enhanced angiogenesis and melanoma growth via α9ß1 integrin.

Tumor-associated macrophages (TAMs) have multifaceted roles in tumor development, particularly linked with tumor angiogenesis and invasion, but the molecular mechanism underlying this association remains unclear. In this study, we report that lack of osteopontin (OPN) suppresses melanoma growth in opn(-/-) mice and macrophages are the crucial component responsible for OPN-regulated melanoma growth. In tumor microenvironment, OPN activates macrophages and influences angiogenesis by enhancing cyclooxygenase-2 (COX-2)-dependent prostaglandin E2 (PGE2) production in an autocrine manner. Furthermore, we identify α9ß1 integrin as a functional receptor for OPN that mediates its effect and activates ERK and p38 signaling, which ultimately leads to COX-2 expression in macrophages. The major role played by OPN and PGE2 in angiogenesis are further amplified by upregulation of MMP-9. OPN-activated macrophages promote the migration of endothelial and cancer cells via PGE2. These findings provide evidence that TAMs serve as source of key components such as OPN and COX-2-derived PGE2 and MMP-9 in melanoma microenvironment. Clinical specimens analyses revealed that increased infiltration of OPN-positive TAMs correlate with melanoma growth and angiogenesis. These data provide compelling evidence that OPN and COX-2 expressing macrophages are obligatory factors in melanoma growth. We conclude that OPN signaling is involved in macrophage recruitment into tumor, and our results emphasize the potential role of macrophage in modulation of tumor microenvironment via secretion of OPN, PGE2 and MMP-9, which trigger angiogenesis and melanoma growth. Thus, blockade of OPN and its regulated signaling network provides unique strategy to eradicate melanoma by manipulating TAMs.

Peak expiratory flow rate of normal school going children in Mymensingh municipality.

This cross sectional study was conducted to measure the PEFR values of normal school going children of Mymensingh, Bangladesh. Peak expiatory flow rate (PEFR) is a lung function test which is easily measurable and reproducible but the base line values of PEFR have not been studied in large scale among Bangladeshi children. In Mymensingh municipality out of 79 schools 4 schools were selected purposively. A total of 984 (499 boys and 485 girls, nearly equal in ratio) normal children (6-15 years), were selected randomly by using lottery technic in classroom. To obtain peak expiratory flow rate (PEFR) mini Wright peak flow meter was used. The highest of three readings was taken as the correct value. Anthropometric parameters including body weight and height were recorded by bathroom scale and stadiometer respectively and body surface area was calculated. According to age interval (10 months) in boys and girls the mean difference of PEFR values showed that among age categories of 6, 7 & 8 years had no significant (p were <0.18, <0.73 and <0.70 accordingly) and all other age categories the mean difference of PEFR between boys and girls had significant (p<0.01). The correlation coefficient (r values) and the level of significance between different anthropometric parameters and PEFR in case of boys and girls were significant (p<0.001). Correlation of height (boys r=0.961 & girls r=0.954) with PEFR was the highest in comparison to other anthropometric parameters (age, sex and body weight). Boys had significantly higher values of PEFR than the girls at any height.

Sexual and urinary outcome after surgical treatment for low rectal cancer.

Carcinoma involving the lower part of the rectum is now successfully managed by sphincter saving surgery with less morbidity and uneventful recovery. This study was designed to observe the sexual and urinary dysfunction in both sexes of the patients suffering from cancer of the lower third of the rectum managed by surgical intervention with preservation of sphincter. A comparative study was carried out on 54 patients with low rectal cancer who underwent ultra-low anterior resection in the department of surgery, Bangabandhu Sheikh Mujib Medical University, Dhaka from January, 2009 to December, 2010. Patients were divided into two groups depending on the tumor distance from anal verge. Thirty one (57%) patients were in Group A (Experimental) where tumor distance was 5cm from anal verge and upper 1cm of anal sphincter was sacrificed during surgical intervention. Twenty three (43%) patients were in Group B (Control) where tumor distance was 6cm from anal verge and whole length (4cm) of anal sphincter was preserved during surgical intervention. The mean±SD age of the patients was 45.96±14.41 years. During surgery, ultra low anterior resection was performed to remove the tumor in all patients and for anastomosis double stapling technique was performed in 52(96%) patients and hand sewn technique was performed in 2(4%) patients irrespective of tumor distance from anal verge. Covering ileostomy was fashioned in all but one patient. During post-operative follow up Sexual activity in both groups of male patients (Potency, P=0.17; ejaculation; satisfaction and loss of libido, P=0.15) and in female patients (Satisfaction and loss of libido, P=0.15) was not significantly hampered following surgery. Urinary function was assessed by incontinence, increased frequency (P=0.54) and retention (n=0) which were not impaired significantly following surgery. Sexual and urinary function was not significantly impaired in both sexes after low rectal cancer surgery. Sphincter saving surgery can be performed in a very low rectal cancer with preservation of anal sphincter.

Chromium (VI) induced acute toxicity and genotoxicity in freshwater stinging catfish, Heteropneustes fossilis.

Acute toxicity and genotoxicity of Chromium(VI) as K2Cr2O7 were evaluated in freshwater stinging catfish, Heteropneustes fossilis. Fish showed behavioral alterations after K2Cr2O7 exposure and 96h-LC50 was 35.724mg/L in semi-static bioassay. Fish were exposed to three sub-lethal concentrations (concentration I=1/4th of 96h-LC50, concentration II=1/10th of 96h-LC50 and concentration III=environmental concentration of Cr reported in the river Buriganga). Blood, liver and gill samples were collected after 48h, 96h and 192h. Micronucleus (MN) assay was conducted in blood erythrocytes and DNA damage was evaluated by comet assay in whole blood, gill and liver tissues. Cr(VI) significantly (p<0.05) induced MN frequency and tail DNA (percent) which increased in a concentration depended manner in all types of tissues. Frequency of MN and tail DNA (percent) increased after 48 and 96h of exposure which decreased after 192h of exposure. The liver was the most sensitive to chromium (VI) exposure among the tissues with highest tail DNA (33.70±0.68 percent) at 9.0mg/L after 96h. This study found MN and comet assays in combination as an adequate approach for ecotoxicological monitoring and Cr(VI) as potential genotoxic agent.

Andrographolide inhibits osteopontin expression and breast tumor growth through down regulation of PI3 kinase/Akt signaling pathway.

Breast cancer is one of the most common cancers among women in India and around the world. Despite recent advancement in the treatment of breast cancer, the results of chemotherapy to date remain unsatisfactory, prompting a need to identify natural agents that could target cancer efficiently with least side effects. Andrographolide (Andro) is one such molecule which has been shown to possess inhibitory effect on cancer cell growth. In this study, Andro, a natural diterpenoid lactone isolated from Andrographis paniculata has been shown to inhibit breast cancer cell proliferation, migration and arrest cell cycle at G2/M phase and induces apoptosis through caspase independent pathway. Our experimental evidences suggest that Andro attenuates endothelial cell motility and tumor-endothelial cell interaction. Moreover, Andro suppresses breast tumor growth in orthotopic NOD/SCID mice model. The anti-tumor activity of Andro in both in vitro and in vivo model was correlated with down regulation of PI3 kinase/Akt activation and inhibition of pro-angiogenic molecules such as OPN and VEGF expressions. Collectively, these results demonstrate that Andro may act as an effective anti-tumor and anti-angiogenic agent for the treatment of breast cancer.

Osteopontin stimulates tumor growth and activation of promatrix metalloproteinase-2 through nuclear factor-kappa B-mediated induction of membrane type 1 matrix metalloproteinase in murine melanoma cells.

Matrix metalloproteinases (MMPs) degrade the extracellular matrix (ECM) and play critical roles in tissue repair, tumor invasion, and metastasis. MMPs are regulated by different cytokines, ECM proteins, and other factors. However, the molecular mechanisms by which osteopontin (OPN), an ECM protein, regulates ECM invasion and tumor growth and modulates MMP activation in B16F10 cells are not well defined. We have purified OPN from human milk and shown that OPN induces pro-MMP-2 production and activation in these cells. Moreover, our data revealed that OPN-induced membrane type 1 (MT1) MMP expression correlates with translocation of p65 (nuclear factor-kappaB (NF-kappaB)) into the nucleus. However, when the super-repressor form of IkappaBalpha (inhibitor of NF-kappaB) was transfected into cells followed by treatment with OPN, no induction of MT1-MMP expression was observed, indicating that OPN activates pro-MMP-2 via an NF-kappaB-mediated pathway. OPN also enhanced cell migration and ECM invasion by interacting with alpha(v)beta(3) integrin, but these effects were reduced drastically when the MMP-2-specific antisense S-oligonucleotide was used to suppress MMP-2 expression. Interestingly, when the OPN-treated cells were injected into nude mice, the mice developed larger tumors, and the MMP-2 levels in the tumors were significantly higher than in controls. The proliferation data indicate that OPN increases the growth rate in these cells. Both tumor size and MMP-2 expression were reduced dramatically when anti-MMP-2 antibody or antisense S-oligonucleotide-transfected cells were injected into the nude mice. To our knowledge, this is the first report that MMP-2 plays a direct role in OPN-induced cell migration, invasion, and tumor growth and that demonstrates that OPN-stimulated MMP-2 activation occurs through NF-kappaB-mediated induction of MT1-MMP.

Insight into the physiological function(s) of uteroglobin by gene-knockout and antisense-transgenic approaches.

To determine the physiological function(s) of uteroglobin (UG), a steroid-inducible, homodimeric, secreted protein, we have generated transgenic mice that either are completely UG-deficient due to UG gene-knockout (UG-KO) or are partially UG-deficient due to the expression of UG antisense RNA (UG-AS). Both the UG-KO and UG-AS mice develop immunoglobulin A (IgA) nephropathy (IgAN), characterized by microhematuria, albuminuria, and renal glomerular deposition of IgA, fibronectin (Fn), collagen, and C3 complement. This phenotype of both UG-KO and UG-AS mice is virtually identical to that of human IgAN, the most common primary glomerulopathy worldwide. The molecular mechanism by which UG prevents this disease in mice appears to center around UG's interaction with Fn. Since Fn, IgA, and UG are present in circulation and high plasma levels of IgA-Fn complex have been reported in human IgAN, we sought to determine whether UG interacts with Fn and prevents Fn-Fn and/or IgA-Fn interactions, essential for abnormal tissue deposition of Fn and IgA. Our coimmunoprecipitation studies uncovered the formation of Fn-UG heteromers in vitro and these heteromers are detectable in the plasma of normal mice, but not UG-KO mice. Further, high plasma levels of IgA-Fn complex, a characteristic of human IgAN patients, were also found in UG-KO mice. Finally, coadministration of UG + Fn or UG + IgA to UG-KO mice prevented glomerular deposition of Fn and IgA, respectively. Our results define a possible molecular mechanism of IgAN and provide insight into at least one important physiological function of UG in maintaining normal renal function in mice.