Pancreatic tumor microenvironmental acidosis and hypoxia transform gold nanorods into cell-penetrant particles for potent radiosensitization.

Coating nanoparticles with stealth epilayers increases circulation time by evading opsonization, macrophage phagocytosis, and reticuloendothelial sequestration. However, this also reduces internalization by cancer cells upon reaching the tumor. We designed gold nanorods (GNRs) with an epilayer that retains stealth properties in circulation but transforms spontaneously in the acidotic tumor microenvironment to a cell-penetrating particle. We used a customized stoichiometric ratio of l-glutamic acid and l-lysine within an amphiphilic polymer of poly(l-glutamic acid-co-l-lysine), or P(Glu-co-Lys), to effect this transformation in acidotic environments. P(Glu-co-Lys)-GNRs were internalized by cancer cells to facilitate potent in vitro radiosensitization. When administered intravenously in mice, they accumulate in the periphery and core of tumors without any signs of serum biochemical or hematological alterations, normal organ histopathological abnormalities, or overt deterioration in animal health. Furthermore, P(Glu-co-Lys)-GNRs penetrated the tumor microenvironment to accumulate in the hypoxic cores of tumors to potently radiosensitize heterotopic and orthotopic pancreatic cancers in vivo.

Knowledge and attitudes towards HIV/AIDS amongst Kuwait University dental students.

The HIV and AIDS have emerged as complex health threats to the world population. As future dentists, it is pertinent that the dental students have sufficient knowledge and a positive approach towards the disease. Accordingly, the aim of this study was to assess the HIV/AIDS related knowledge and attitudes amongst clinical dental students at Kuwait University. A cross-sectional survey was conducted amongst the clinical dental students using a structured questionnaire with 60 questions to examine their knowledge under various categories and 13 questions to examine their attitudes towards the disease. The survey revealed that almost 58% of the respondents demonstrated a high level of knowledge (mean score: 45.23 ± 4.35 SD). Majority of the students (63.6%) expressed negative attitude (mean score: 5.36 ± 2.56 SD). The mean knowledge score of the fifth year dental students was significantly higher (P = 0.022) than that of the final year dental students regarding the knowledge of virus and disease process. However, no significant difference was observed with respect to other knowledge categories. Despite their high level of knowledge, the majority displayed a negative attitude towards HIV/AIDS. Hence, the findings imply that there is a need to address, more clearly, the students' misconceptions and attitudes towards the disease.

Growth hormone receptor and IGF-I receptor immunoreactivity during orthodontic tooth movement in the prednisolone-treated rat.

Bone remodeling during tooth movement is regulated by local and systemic factors. Two regulators of bone metabolism are growth hormone (GH) and insulin-like growth factor-I (IGF-I). Their effects are mediated via binding to GH receptor (GHR) and IGF-I receptor (IGF-IR) in target tissues. Corticosteroids may affect the activity of these growth factors. This study examined the effect of prednisolone on GHR and IGF-IR expression in dental tissues following orthodontic tooth movement. The corticosteroid-treated group (N = 6) was administered prednisolone (1 mg/kg) daily and the control group (N = 6) received equivalent volumes of saline. An orthodontic force (30 g) was applied to the maxillary first molar. Animals were sacrificed 12 days postappliance insertion. Sagittal sections of the first molar were stained for GHR and IGF-IR immunoreactivity. GHR and IGF-IR cell counts were elevated following appliance-treatment. Orthodontic tooth movement appeared to up-regulate GHR and IGF-IR immunoreactivity, but this up-regulation was reduced following prednisolone treatment. The suppression of GHR and IGF-I immunoreactivity in steroid-treated animals infers the mechanism whereby bone resorption and deposition, necessary for orthodontic tooth movement, may be inhibited by prednisolone. However, at 12 days postappliance insertion, no difference in orthodontic tooth movement was observed following low-dose prednisolone treatment.

Effect of a static magnetic field on orthodontic tooth movement in the rat.

Orthodontic tooth movement may be enhanced by the application of a magnetic field. Bone remodelling necessary for orthodontic tooth movement involves clastic cells, which are tartrate-resistant acid phosphatase (TRAP) positive and which may also be regulated by growth hormone (GH) via its receptor (GHR). The aim of this study was to determine the effect of a static magnetic field (SMF) on orthodontic tooth movement in the rat. Thirty-two male Wistar rats, 9 weeks old, were fitted with an orthodontic appliance directing a mesial force of 30 g on the left maxillary first molar. The appliance incorporated a weight (NM) or a magnet (M). The animals were killed at 1, 3, 7, or 14 days post-appliance insertion, and the maxillae processed to paraffin. Sagittal sections of the first molar were stained with haematoxylin and eosin (H&E), for TRAP activity or immunohistochemically for GHR. The percentage body weight loss/gain, magnetic flux density, tooth movement, width of the periodontal ligament (PDL), length of root resorption lacunae, and hyalinized zone were measured. TRAP and GHR-positive cells along the alveolar bone, root surface, and in the PDL space were counted. The incorporation of a SMF (100-170 Gauss) into an orthodontic appliance did not enhance tooth movement, nor greatly alter the histological appearance of the PDL during tooth movement. However significantly greater root resorption (P = 0.016), increased width of the PDL (P = 0.017) and greater TRAP activity (P = 0.001) were observed for group M at day 7 on the compression side. At day 14 no differences were observed between the appliance groups.

The effect of glucocorticosteroid treatment on dentine formation in the Lewis rat, a histological study.

Glucocorticosteroids are widely used in the treatment of chronic illnesses and have been reported to cause premature obliteration of the pulp space. During the active stages of dentinogenesis, odontoblasts are growth hormone receptor (GHr) positive. The aims of this study were to determine if the glucocorticosteroid, prednisone, affected the rate of dentine deposition and odontoblast expression of GHr in the rat molar. Following subcutaneous injection of 0.05 mg/kg, 1.0 mg/kg or 5.0 mg/kg prednisone for 20 days, immature and mature molars from rats aged 3 and 6 weeks respectively, were examined histologically. Distribution of GHr expression was determined immunohistochemically. No morphological differences were observed in molars from prednisone treated animals. Prednisone did not appear to enhance dentine deposition in immature molars but in mature molars significantly increased dentine deposition on the roof of the pulp chamber at a dosage of 5.0 mg/kg (p < 0.001). In all immature molars, odontoblasts and pulp cells expressed GHr immunoreactivity. In mature molars, odontoblasts and pulpal cells from controls did not show GHr immunoreactivity. However, odontoblasts and pulp cells were GHr immunoreactive in mature molars from animals treated with prednisone.

Insulin-like growth factor-I (IGF-I) and IGF-I receptor (IGF-IR) immunoreactivity in normal and osteopetrotic (toothless, tl/tl) rat tibia.

Insulin-like growth factor-I (IGF-I) plays a major role in regulating cell growth. This study examined the immunohistochemical distribution of IGF-I and IGF-I receptor (IGF-IR) in tibias from normal and osteopetrotic (toothless, tl/tl) rats, following treatment with colony stimulating factor-1 (CSF-1). In normal rats, immunoreactivity for IGF-I and IGF-IR was detected in cells of the articular and epiphyseal cartilage, secondary ossification centres, zones of resting and proliferating chondrocytes and bone marrow. Bone marrow cells immunoreactive for IGF-I and IGF-IR were significantly reduced in the tl/tl rat (p < 0.001) compared with normal animals. Treatment of tl/tl rats with CSF-1 increased immunoreactivity for IGF-I and IGF-IR in bone marrow cells as well as the number of TRAP positive osteoclasts. This increase was the result of recruitment of a range of hematopoietic cell types, including eosinophils, polymorphs and a substantial number of monocyte-like cells demonstrating strong immunoreactivity to IGF-I/IGF-IR. The differences in relative immunoreactivity for IGF-I/IGF-IR by bone marrow cells in untreated and CSF-1-treated tl/tl rats indicate a CSF-1-dependent recruitment of cells bearing surface IGF-IRs which may be mediated by an increase in local or systemic IGF-I.

Pelvic arteriovenous malformation mimicking seminal vesicle cyst.

Congenital pelvic arteriovenous malformations are rare entities, especially in males. Presenting symptoms, if any at all, are commonly a mass, thrill, bruit, or pain. Treatment options include surgical extirpation, embolization, or a combination of both. This case provides support for the last option in a patient presenting with symptoms localized to the seminal vesicles.

Cat-scratch disease: a case report.

Cat-scratch disease is the most common cause of benign lymphadenopathy in children and young adults. A 21-year-old patient presented with a 14-day history of asymptomatic left submental swelling. Clinical and radiographic examination revealed no dental cause. An enlarged, firm nodule was excised. The histopathologic appearance was consistent with cat-scratch disease. This article summarizes the relevant literature and discusses diagnosis and treatment of cat-scratch disease.

Tooth movement and vascularity of the dental pulp: a pilot study.

The effect of orthodontic tooth movement on the dental pulp was assessed histologically in twelve subjects. The participants in this study required the extraction of at least two maxillary first premolars for orthodontic treatment. They were asked to wear a maxillary removable appliance that acted to move a randomly determined premolar in a buccal direction. The appliance was designed to avoid contacting the contra-lateral tooth that was used as the matched control. The appliance was initially worn for a week to ensure patient comfort and cooperation. The appliance was then activated and the patient dismissed. After two weeks, the appliance was reactivated. Both the control and experimental teeth were extracted three weeks later, on the thirty-fifth day of activated appliance wear. The teeth were fixed, decalcified and sectioned. The sections were stained with haematoxylin and eosin for histological examination. This investigation demonstrated that orthodontic tooth movement did have an effect upon the dental pulp, causing vasodilation in the pulp of an orthodontically stressed tooth.

Ameloblast apoptosis and IGF-1 receptor expression in the continuously erupting rat incisor model.

Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are "hard wired" for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.

Identification of a breast cancer-specific gene, BCSG1, by direct differential cDNA sequencing.

A high-throughput direct-differential cDNA sequencing approach was employed to identify genes differentially expressed in normal breast as compared with breast cancer. Approximately 6000 expressed sequence tags (ESTs) from cDNA libraries of normal breast and breast carcinoma were selected randomly and subjected to EST-sequencing analysis. The relative expression levels of more than 2000 unique EST groups were quantitatively compared in normal versus cancerous breast. Of many putative differentially expressed genes, a breast cancer-specific gene, BCSGC1, which was expressed in high abundance in a breast cancer cDNA library but scarcely in a normal breast cDNA library, was identified as a putative breast cancer marker. In situ hybridization analysis demonstrated stage-specific BCSG1 expression as follows: BCSG1 was undetectable in normal or benign breast lesions, showed partial expression in ductal carcinoma in situ, but was expressed at an extremely high level in advanced infiltrating breast cancer. The predicted amino acid sequence of BCSG1 gene has a significant sequence homology to the non-amyloid beta protein fragment of the Alzheimer's disease amyloid protein. BCSG1 overexpression may indicate breast cancer malignant progression from benign breast or in situ carcinoma to the highly infiltrating carcinoma.

Non-expression of insulin-like growth factor-I receptor is associated with apoptosis: an ultrastructural study on rat ameloblasts.

Insulin-like growth factor-I (IGF-I) is a pleiotrophic polypeptide which appears to have roles both as a circulating endocrine hormone and as a locally synthesized paracrine or autocrine tissue factor. IGF-I plays a major role in regulating the growth of cells in vivo and in vitro and initiates metabolic and mitogenic processes in a wide variety of cell types by binding to specific type I receptors in the plasma membrane. In this study, we report the distribution of IGF-I receptors in odontogenic cells at the ultrastructural level using the high resolution protein A-gold technique. In the pre-secretory stage, very little gold label was visible over the ameloblasts and odontoblasts. During the secretory stage the label was mostly seen in association with the cell membranes and endoplasmic reticulum of the ameloblasts. Lysosome-like elements in the post-secretory stage were labelled as well as multivesicular dense bodies. Very little labelling was encountered in the ameloblasts in the transitional stage, where apoptotic bodies were clearly visible. The maturation stage also exhibited labelling of the secretory-like granules in the distal surface. The presence of gold particles over the plasma membrane is an indication that IGF-I receptor is a membrane-bound receptor. Furthermore, the intracellular distribution of the label over the endoplasmic reticulum supports the local synthesis of the IGF-I receptor. The absence of labelling over the transitional ameloblasts suggests that the transitional stage may require the non-expression of IGF-I as a prerequiste or even a trigger for apoptosis.

The glandular odontogenic jaw cyst: report of a case.

A case of a rare odontogenic cyst arising in the lateral periodontal membrane in the mandible in a 14 year old girl is reported. This lesion appeared to be a new entity and has been named glandular odontogenic cyst (GOC) or sialo-odontogenic cyst. Histologically the lesion was lined by mucous producing cuboidal epithelium containing several areas of thickening and numerous duct-like structures. The cyst recurred with the same histology two years postoperatively.

In situ hybridization evidence for a paracrine/autocrine role for insulin-like growth factor-I in tooth development.

Insulin-like growth factor-I(IGF-I) has both metabolic and growth-promoting activities in many cell and tissue types. Although IGF-I is present in serum, it is also thought to have important autocrine and paracrine functions. Immunohistochemistry for IGF-I and its receptor have shown that IGF-I is synthesised locally by the tooth forming cells which exhibit both the IGF-I and the growth hormone receptors. This concept required to be tested by in situ hybridization. Using a digoxigenin-labelled synthetic oligodeoxyribonucleotide probe for IGF-I, we investigated the distribution of IGF-I mRNA in the continuously erupting rat incisor by in situ hybridization. The distribution and intensity of the hybridization signal varied with the developmental stage of the rat incisor. The cells of the apical loop expressed a positive hybridization signal, but the earliest polarised odontoblasts and pre-ameloblasts did not show any positive signal. The onset of enamel secretion was accompanied by a strong hybridization signal in the secretory ameloblasts as well as the odontoblasts. Maturation ameloblasts also demonstrated IGF-I message in their cytoplasm as well as their nuclei. The cells of the pulp and the dental follicle were consistently negative. However, in the adjacent alveolar bone, the signal was high in the osteoblasts and osteoclasts. These findings support the notion of paracrine or autocrine function for IGF-I in tooth development.

Review article: The role of oncogenes, tumour suppressor genes and growth factors in oral squamous cell carcinoma: a case of apoptosis versus proliferation.

Mutation, deactivation and disregulated expression of oncogenes and tumour-suppressor genes may be involved in the pathogenesis of oral squamous cell carcinoma (SCC). Deactivation of the p53 tumour-suppressor gene allows cell proliferation and blocks apoptosis of malignant oral keratinocytes. Mutation in the ras oncogene results in persistent mitogenic signalling. Upregulatioed c-Myc expression, in the presence of growth factors, provides an additional proliferative signal. Loss of retinoblastoma tumour-suppressor gene (Rb) function may contribute to oral keratinocyte hyperproliferation and recent evidence suggests that simultaneous deactivation of both p53 and Rb is required for tumourigenesis. Enhanced Bcl-2 and reduced Fas expression inhibit tumour cell apoptosis and may convey resistance to cytotoxic drugs and T cell-mediated cytotoxicity, respectively. Exogenous mutagens such as tobacco, alcohol and viral oncogenes may cause altered expression of oncogenes and tumour-suppressor genes in some cases of oral SCC. The impact of these mechanisms on future therapies for oral SCC is highlighted.

Expression and localization of sulphated glycoprotein-2 mRNA in the rat incisor tooth ameloblasts: relationships with apoptosis.

The expression of sulphated glycoprotein-2 (SGP-2) is associated with the onset of cellular atrophy and death in many rodent tissues. This gene has a multifunctional involvement that includes apoptosis, spermatogenesis, promotion of cell-cell interactions, modulation of complement systems and tissue regeneration and remodelling. Using decalcified mandibles, mRNA for SGP-2 in rat incisor tooth ameloblasts was examined by in situ hybridization using 35S riboprobes. The rat incisor is unique in that, at one time, all stages of the complex life cycle of the ameloblasts are represented along the length of the enamel forming aspect of the tooth. The pre-ameloblasts only secrete enamel matrix after mitosis. When the full thickness of the enamel has been formed, a remarkable transition in phenotype takes place in the ameloblast. This transition is accompanied by apoptosis or programmed cell death of approximately 25% of ameloblasts. An additional 25% of ameloblasts undergo apoptosis when maturation of enamel matrix takes place with removal of water and protein from the increasingly mineralized matrix. In the present study, expression of SGP-2 was localized most often in the post-secretory transition and maturation ameloblasts. In contrast, the presecretory and secretory ameloblasts did not demonstrate specific hybridization signals. Consistently, neither the odontoblasts nor the pulp demonstrated hybridization signals. Hence our results support other published results which show that increased expression of SGP-2 is associated with apoptosis. The exact function of the SGP-2 gene and its products is not fully defined.(ABSTRACT TRUNCATED AT 250 WORDS)

Prenatal expression of growth hormone receptor/binding protein and insulin-like growth factor-I (IGF-I) in the enamel organ. Role for growth hormone and IGF-I in cellular differentiation during early tooth formation?

Immunohistochemistry was used to study the ontogeny of GH receptor/binding protein (GHR/BP) and IGF-I from the 13-day-old embryo (E13) to the E19 rat fetus in the developing incisor and molar. Analysis of serial sections revealed diffuse staining of GHR/BP and IGF-I at the bud and early cap stages within both the mesenchyme of the dental papilla and the ectodermal-derived enamel organ. Just before transition to the cap stage, immunoreactivity of GHR/BP and IGF-I increased in the epithelial bud and extended to the condensed dental mesenchyme. At the cap stage, the dental epithelium showed an intense expression of GHR/BP and IGF-I, whereas the dental mesenchymal cells showed very weak staining. The inner enamel epithelium and the outer enamel epithelium were positive for both GHR/BP and IGF-I in the bell stage. Differentiating ameloblasts, odontoblasts and the secretory ameloblasts and odontoblasts continued to express GHR/BP and IGF-I in incisors. These findings support the premise that growth hormone and IGF-I may play a role in embryonic tooth development by regulating the epithelial-mesenchymal interactions that influence events in growth and cytodifferentiation.

Insulin-like growth factor-I receptor in the cell biology of the ameloblast: an immunohistochemical study on the rat incisor.

The distribution of IGF-I receptor is reported in the odontogenic epithelium and mesenchyme of the continuously erupting mandibular incisor of the rat by immunohistochemistry using a polyclonal antibody specific to the IGF-I receptor. Odontogenic epithelium is a unique odontogenic sequence in that all stages of the complex life cycle of the ameloblast are represented along the length of the enamel-forming aspect of the tooth. Pre-ameloblasts become post-mitotic before secreting enamel matrix. When the full thickness of the enamel has been formed, a remarkable transition in phenotype takes place in the ameloblast. It changes from a protein secretory cell to one active in maturation of enamel matrix by removal of water and protein from the increasingly mineralized matrix. The distribution and intensity of IGF-I receptor expression varied with the phenotypic stages of the ameloblasts. Diffuse cellular staining for IGF-I receptor was found during the active secretory phase of amelogenesis. However, towards the end of this phase, the staining was confirmed to granular or vesicular structures within the cytoplasm. These granular deposits gradually decreased as the ameloblasts made the transition towards enamel maturation. This transition is accompanied by programmed cell death (apoptosis) of approximately 25% of the ameloblasts and cells in this zone did not stain for IGF-I receptor. With the onset of enamel maturation, diffuse staining of the ameloblast layer was re-established gradually and staining remained evident right up to the reduced enamel epithelium, which joins with the oral epithelium. Strong IGF-I receptor immunoreactivity was observed in the stratum basale and stratum spinosum of the adjacent labial gingival epithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression and regulation of insulin-like growth factor-I in the rat incisor.

Growth factors play an important role in the regulation of cell growth, division and differentiation. In this study the distribution and regulation of insulin-like growth factor-I (IGF-I) in the continuously erupting rat incisor was determined by immunohistochemistry. Results were evaluated both visually and with a computer-based image analysis system. The distribution and intensity of IGF-I immunoreactivity varied with developmental stage of the rat incisor. Strong IGF-I immunoreactivity was observed in differentiating odontoblasts and ameloblasts. The most intense immunoreactivity was observed in secretory ameloblasts, secretory odontoblasts and in maturation ameloblasts. Staining was weak or absent in post-secretory ameloblasts but persisted in post-secretory odontoblasts. Weak to moderate immunoreactivity was also seen in cells of the stratum intermedium and in the reduced enamel epithelium. Surrounding alveolar bone showed strong IGF-I immunoreactivity in osteoblasts and in the stratum basale and stratum spinosum of the adjacent labial gingival epithelium. In order to assess the role of GH in IGF-I expression, GH (65 micrograms/100 g bw) was administered for six days to dwarf GH deficient rats, producing a significant increase in body weight (P < 0.01). Measurements at different stages of odontogenesis showed that the staining intensity of secretory ameloblasts (P < 0.01) and maturation ameloblasts (P < 0.001) was significantly different between untreated and treated animals. These results indicate that IGF-I is present in cell populations of the enamel organ of the rat incisor found previously to exhibit growth hormone receptors, and that expression of IGF is GH dependent.(ABSTRACT TRUNCATED AT 250 WORDS)