A laboratory study to test the responses of human dental pulp stem cells to extracts from three dental pulp capping biomaterials.

AIM: This laboratory study aimed to investigate the effects of three endodontic biomaterials; MTA-HP, iRoot-BP-Plus and ACTIVA on the proliferation, adhesion and osteogenic differentiation of human Dental Pulp Stem Cells (hDPSCs). METHODOLOGY: The hDPSCs were isolated from the dental pulps of 21 patients scheduled for surgical extraction of their impacted third molars. The MTT assay was used for assessing cellular proliferation. Ninety-six-well plates were used and the experiment was repeated four times under the same condition and the assay was done in triplicate. Four groups were assigned in which the hDPSCs were cultured in complete media only and considered as negative control. Whilst in the 2nd , 3rd and 4th groups, the cells were treated with CM supplemented with 1.5 µl MTA-HP (CM-MTA, iRoot-BP-Plus (CM-BP), and ACTIVA(CM-AC) extracts, respectively. Attachment adhesion and growth morphology of hDPSCs were observed using SEM and the osteogenic differentiation assay was evaluated by Alizarin red stain test (ARS). The data of proliferation and osteogenic differentiation were analysed using two-way ANOVA followed by Tukey's post hoc multiple comparison test. A p-value < 0.05 was considered significant to analyse the differences amongst the means of groups. RESULTS: Both CM-MTA and CM-BP groups were associated with a significant increase in hDPSC proliferation in comparison with CM-AC and CM groups (p = 0.001). hDPSCs exhibited a greater cellular attachment to iRoot-BP-Plus surfaces followed by MTA-HP, whilst less attachment was observed in the ACTIVA group. Moreover, at day 7 there was a significant difference in formation of mineralizing nodules; CM-BP, CM-MTA and CM-AC groups respectively (p = 0.001). Whilst there was no significance of difference between CM-AC and CM groups (p > 0.05). CONCLUSIONS: In a laboratory setting, ACTIVA, MTA-HP and iRoot-BP-Plus promoted hDPSCs proliferation, mineralization and attachment, which may explain their in-situ success as endodontic biomaterials.

Tuberculosis is a Mimicker of JIA: A Rare Case Report.

Cystic tuberculosis of the bone is a rare form of tuberculosis (TB). The condition presents like Juvenile idiopathic arthritis (JIA) of children. In children, the lesions symmetrically involve the peripheral skeleton, which are less sclerotic than adults. A case report is presented here where the patient presented with i) the extensive involvement of bones with cystic lesion, ii) Hand & feet involvement with multiple bony exostosis iii) Synovial swelling of multiple joints and 4) fever for 6 months. Swelling of the joints was disproportionately greater than pain. For the last 6 month patient developed low grade fever with evening rise of temperature & dry cough. There was associated anorexia & significant weight loss. Patient was moderately anaemic & there was cervical lymphadenopathy on both sides. The chest examination revealed features suggestive of consolidation in the right lung. Regarding MSK findings there was swelling of both knee & right wrist with G-II tenderness, sublaxation of both anterior and posterior cruciate ligament with mild effusion. Bony exostosis at the base of left index finger & at the base of right middle finger was found. Investigation shows low Hb, very high ESR, positive tuberculin test. X-ray Chest suggestive of consolidation, FNA of right cervical lymph node consistent with tuberculosis. X-ray Pelvis has shown expansile mixed sclerotic radiolucent areas with interval septation involving upper part of both femoral shafts. MRI findings of right knee joint were suggestive of tuberculous osteomyelitis. At this stage the patient was put on Anti-TB chemotherapy. After 2 month and 4 month of follow up with Anti-TB drug both MSK & lung condition was improved significantly. Ultimately the patient was diagnosed as cystic tuberculosis of bone & continued Anti-TB drugs.

Diagnostic Accuracy of High-Resolution 3D T2-SPACE in Detecting Cerebral Venous Sinus Thrombosis.

BACKGROUND AND PURPOSE: Assessment of cerebral venous sinus thrombosis on MR imaging can be challenging. The aim of this study was to evaluate the diagnostic accuracy of high-resolution 3D T2 sampling perfection with application-optimized contrasts by using different flip angle evolution (SPACE) in patients with cerebral venous sinus thrombosis and to compare its performance with contrast-enhanced 3D T1-MPRAGE. MATERIALS AND METHODS: We performed a blinded retrospective analysis of T2-SPACE and contrast-enhanced MPRAGE sequences from patients with cerebral venous sinus thrombosis and a control group. The results were compared with a reference standard, which was based on all available sequences and clinical history. Subanalyses were performed according to the venous segment involved and the clinical stage of the thrombus. RESULTS: Sixty-three MR imaging examinations from 35 patients with cerebral venous sinus thrombosis and 51 examinations from 40 control subjects were included. The accuracy, sensitivity, and specificity calculated from the initial MR imaging examination for each patient were 100% each for T2-SPACE and 95%, 91%, and 98%, respectively, for contrast-enhanced MPRAGE. The interrater reliability was high for both sequences. In the subanalysis, the accuracy for each venous segment involved and if subdivided according to the clinical stage of thrombus was ≥95% and ≥85% for T2-SPACE and contrast-enhanced MPRAGE, respectively. CONCLUSIONS: Both T2-SPACE and contrast-enhanced MPRAGE offer high accuracy for the detection and exclusion of cerebral venous sinus thrombosis; however, T2-SPACE showed a better overall performance and thus could be a useful tool if included in a multiparametric MR imaging protocol for the diagnosis of cerebral venous sinus thrombosis, especially in scenarios where gadolinium administration is contraindicated.

Tuberculosis of knee joint and tubercular pyomyositis of gastrocnemius muscle.

Extra-spinal musculoskeletal tuberculosis (TB) is rare and tubercular pyomyositis is extremely rare. Tuberculosis of the knee-joint is a haematogenous infection secondary to a focus of active disease elsewhere in the body which may not be found. Tubercular pyomyositis usually caused by invasion from the adjacent structures rather than a secondary spread. Here we describe a 40 years old male patient who presented to us with pain in the right knee joint for one and half years and pain and swelling of right calf muscle for fifteen days. He was eventually diagnosed as a case of tuberculosis of the right knee joint and tubercular pyomyositis of right gastrocnemius muscle on the basis of fine needle aspiration from the right calf that showed caseation necrosis with clusters of epithelial cells despite absence of systemic symptoms, the absence of other foci of active tuberculosis and a normal chest radiograph.

Susceptibility-weighted MR imaging for diagnosis of capillary telangiectasia of the brain.

BACKGROUND AND PURPOSE: BCT is a benign entity, whose appearance on conventional MR imaging makes its differentiation from neoplastic, inflammatory, or subacute ischemic disease challenging. SWI is sensitive to susceptibility effects from deoxyhemoglobin with excellent spatial resolution. Only scarce case reports have described the utility of SWI in cases of BCT. Our aim was to show the diagnostic value of SWI applied to a larger series of cases. MATERIALS AND METHODS: This was an observational retrospective study of 33 BCTs in 27 consecutive patients examined from August 2009 to January 2011 with MR imaging, including SWI. Morphology, signal intensity characteristics, and additional vascular malformations were analyzed. Preceding or follow-up examinations were available in 18 patients with a median time interval of 14.5 months (range, 2-115 months). RESULTS: Twenty-five pontine and 8 supratentorial BCTs demonstrated distinct signal-intensity loss on SWI in combination with postcontrast enhancement. Mean lesion diameter was 4.9 mm (range, 1.5-17 mm). Thirty-nine percent showed slight signal-intensity changes on T1 and/or T2; the remainder were isointense to normal brain. In 30%, a prominent draining vessel was observed. Additional cerebral vascular malformations were found in 5 patients. CONCLUSIONS: SWI represents a valuable tool for confirmation of presumed BCT. Demonstration of signal-intensity loss on SWI in an enhancing focal brain lesion, otherwise unremarkable on conventional MR images, is highly specific for BCT, thus excluding serious pathology and reassuring the patient and referring physician. This is particularly helpful for BCT in less typical locations.

Differentiation of male chick embryo gonads during the last week of incubation in organ and tissue culture.

15th day chick embryo testes, were cultivated in organ and tissue culture for 5 days, in semisynthetic media. The histological examination of the cultured testes shows a noticeable retardation in development compared with normal testes at the same age. Microscopic examination of cultured testicular cells, shows two types of cells, epithelial and fibroblasts. The most remarkable result, the absence of germ cells from the tissue culture.

The effect of adrenocorticotropic hormone (ACTH) on embryonic chicken gonads.

Chicken embryos were exposed to a single dose of adrenocorticotropic hormone (ACTH) on the 15th day of the incubation period. Five days later, the female embryos showed a significant increase in ovarian weight, ovarian thickness and oocyte count. In contrast, the male embryos showed a significant decreased in testicle weight, seminiferous cord diameter and gonocyte number. The opposite response of testes and ovaries is discussed.

The overlapping effects of thyrotropin and gonadotropins on chick embryo gonads in vitro.

Pieces of 12- and 15-day-old chick embryo testes and ovaries were cultured in vitro in the presence of thyrotropin (TSH), gonadotropins (FSH + LH) and adrenocorticotropin (ACTH) for different periods. All the explants of treated gonads differentiated into typical testes or ovaries according to their genetic sex. The gonads of 12-and 15-day-old chick embryos showed a good response to both thyrotropic and gonadotropic stimulation. On the other hand, they did not respond to adrenocorticotropic stimulation. Fifteen-day-old chick embryo testes were grown in tissue culture in the presence of the said hormones. Gonadotropins and TSH enhanced the growth and migration of testicular cells as compared with the control or ACTH treated group. In addition, they maintained the germ cells on the upper surface of epithelial cells. These results have confirmed our previous results in vivo in that gonadotropins and thyrotropin hormones accelerated the development of 12- or 15-day-old chick embryo gonads.

Electron microscopic study of the overlapping effect of thyrotropin and gonadotropins on the Sertoli cells of chick embryo.

The ultrastructural features of the Sertoli cells of the embryonic chick testis have been studied following the administration of thyrotropin (TSH) and gonadotropins (FSH + LH), on the 8th or 15th day of embryonic life. The principal changes observed in the 15-day TSH-treated embryos were the increased quantities of Sertoli cell organelles, particularly the agranular endoplasmic reticulum, mitochondria and Golgi complex. Although the same changes were observed in the gonadotropins-treated embryos, it is worthwhile to mention that the Sertoli cells of 15-day TSH-treated embryos contain abundant granular endoplasmic reticulum more than that in the gonadotropin treated ones. The most striking ultrastructure feature, the occurrence of coated vesicles in Sertoli cells in all groups including control, however they appear much numerous in 15-day treated embryos. The findings of the present investigation support the hypothesis that TSH has an extrathyroidal role and demonstrate the functional responsiveness of 15-day chick embryo Sertoli cells to this hormone. The FSH-like activity of TSH agrees with our earlier hypothesis: namely that hormones of similar molecular structure may bind to the same binding sites as a consequence of receptor immaturity.

Impact of a single neonatal gonadotropin (FSH + LH) or thyrotropin (TSH) treatment on the sexual behaviour of the adult male rat.

A single gonadotropin (FSH + LH) treatment of neonatal male rats resulted in depression of sexual activity in adulthood. It appears that not only steroids, but also gonadotropins may alter adult sexual behaviour by a single neonatal exposure. The chemically related hormone thyrotropin (TSH) had a similar, but much less pronounced, effect on adult sexual activity.

Effect of combined gonadotropin-thyrotropin treatment on development of testis and ovarium in the chickling.

Both TSH and gonadotropins stimulate the gonads in the perinatal period. Gonadotropins influence primarily the parenchyma and they induce an increase in the diameter of seminiferous tubules, and of the thickness of the ovarian cortex. TSH acts primarily on the interstice, but it acts also on the parenchyma and this action is more pronounced in the testis than in the ovary. The hormones can replace each other, and gonadotropins enhance the effect of TSH on the interstice.

Effect of gonadotropins and thyrotropin on the testes and ovaries of the newly hatched chicken.

In newly hatched male and female chickens thyrotropin (TSH) and gonadotropins (FSH, LH) were found to increase the number of spermatogonia or Sertoli cells in the seminiferous cord, and the number of granulosa cells in the ovarian follicle. The effect of gonadotropins was more pronounced than that of TSH. Regarding the overlapping effect of thyrotropin and gonadotropins, in both male and female neonatal gonads the hormones similar in structure seem to be able to substitute each other. They do not however, change each other's effect on the parenchymatous tissue (number of spermatogonia and Sertoli cells in the male, and number of follicles in the female) if they are administered one after the other.

The overlapping effect of gonadotropins and TSH on embryonic chicken gonads.

Determination of gonadal weight, seminiferous canalicule diameter, thickness of ovarian cortex and medulla, as well as of gonadal cell counts in 20-day chicken embryos exposed to gonadotropin (FSH and LH) or TSH at 15 days of pre-hatching life has shown that the examined hormones were able to evoke gonadal response also prenatally. TSH was found to overlap gonadotropin effect in causing increase of seminiferous canalicule diameter, testicular weight and gonocyte counts. In respect of the latter two parameters TSH was even more active than an identical dose of gonadotropin. This indicates that the interstitial and germinal cell receptors of the embryonic testis are not able to differentiate between chemically related, but - in adulthood - functionally different hormones. The ovarian effect of TSH was significant in respect of enhancing organ weight and oocyte mitosis, but not significant for the other parameters.

Histological analysis of the overlapping effect of hypophyseal hormones on the cockerel's testes.

Histological examination of cockerel testes revealed the overlapping effects of FSH and TSH. Within the dose-range studied (20-160 microgram FSH and 27.5-220 microgram TSH), both hormones increased the cell number in the seminiferous cords including the number of spermatogonia, primary spermatocytes and pre-Sertoli cells. They also enhanced the mitotic activity of spermatogonia and accelerated spermatogenesis. Peak effects were observed after treatment with 160 microgram FSH and 55 microgram TSH dose. Liquefaction of cords due to hormone treatment was indicative of an acceleration of testicular ontogeny. Cross-effects of the two hormones were explained by receptor immaturity i.e. in the early stage of ontogenesis the receptors can bind both hormones due to the similarities in their structure. The maximum effects of the hormones were different, that of FSH being more marked.

Overlapping effect of follicle stimulating hormone (FSH) and thyrotropin (TSH) on the ultrastructure of immature chicken testes.

In newly hatched chicken testes the gonadotropin receptors due to their immaturity are not specific but still structurally versatile and so they can bind both FSH and TSH which have a chemically related structure. The functional overlapping effect of FSH and TSH on the ultrastructure of Sertoli and Leydig cells of immature chicken testes was investigated. The activity of Sertoli cells was increased by FSH treatment and this increase correlated well with the amount of SER and RER in cells and with their increased surface activity. The vacuolization and degeneration observed at the apical part of the cells may refer to the formation of testicular tubules. After TSH treatment cell activity increased and in addition a considerable increase in RER and lipid droplets was observed. Fenestrated cisternae were often found in the Sertoli cells of the treated animals. In the Leydig cells, both hormones increased lysosomal activity and the number of lipid droplets. After FSH treatment the amount of SER increased while after TSH treatment the Golgi activity.

Effects of mammalian hypophyseal hormones: gonadotropins, thyrotropin and adrenocorticotropin, on the embryonic development of 8-days chick embryo gonads.

When injected into chick eggs on the 8th day of incubation, gonadotropins (FSH + LH) were shown to stimulate the gonadal development on the 20th day of embryogenesis in both sexes. An increase in the testicular weight, in the seminiferous cord diameter as well as in germ and Sertoli cell number was observed in males. Similarly an increase in ovarian weight, in ovarian width, in cortical thickness and in the number of oocytes was reported in females. By contrast, thyrotropin (TSH) and adrenocorticotropin (ACTH) caused a noticeable decrease in gonadal development in both males and females as compared to the controls or to the gonadotropin-treated groups. It is concluded that the gonads of 8 days chick embryo have the capacity to respond to gonadotropic stimulation and not to the other hypophyseal hormones (TSH or ACTH). This suggests the possibility that at this early stage (8 days of incubation) the external specific hormone could induce its future hormone receptors.

Permanent amplifying effect on the cockerel gonad of hypophysis hormones (FSH, TSH) administered at hatching.

The effect of TSH and FSH was studied by quantitative histological analysis of the gonads in five-week-old cockerels pre-treated with these hormones at hatching. It was concluded that (1) hypophyseal hormones of similar molecular structure are not fully specific in immature animals. At five weeks both FSH and TSH enhance spermatogonium division and spermatogenesis, and increase the number of Sertoli cells. (2) While a single post-hatching dose of hormone has no effect on germinal cells as studied in the fifth week it increases the number of Sertoli cells (3) post hatching FSH-treatment facilitates the effect of hormone treatment at five weeks, amplifying the effect of FSH slightly more than that of TSH. Post hatching TSH-treatment also amplifies the effect of FSH administered at five weeks but does not influence the effect of five-week TSH-treatment on germinal cells. (4) The present results have corroborated earlier findings based on the measurement of gonadal weight and tubule diameter.

The ultrastructure of adrenocortical (interrenal) cells of normal and ACTH-treated chick embryos.

The ultrastructural features of the interrenal cells have been studied in 20-days-old chick embryos following the administration of adrenocorticotropin (ACTH), on the 8th or 15th day of incubation. The interrenal cells of normal embryos contain more SER than RER, mitochondria with tubular cristae, lipid droplets and Golgi complex. Adjacent cells had numerous regions of pentalaminar fusion and intermediate junctions. Neither a precise organization of medullary tissue in relation to interrenal tissue nor any structural differences between interrenal cells were found. Changes in the fine structure of the interrenal cells following ACTH treatment were extensive. The organelles that are known to be involved in the biosynthesis of steroids displayed structural modifications. These were mainly SER, mitochondria, and lipid droplets. Changes were also noticeable in the Golgi complexes, membrane-bound dense bodies (especially in 15 days treated embryos). These results indicate the well developed organelles in the interrenal cells of 20-days-old embryos, and their capacity to respond to ACTH stimulation as early as the 8th day of embryogenesis.

Hepatic fibrosis in hamsters experimentally infected with S. mansoni.

Adult males Schistosoma mansoni were detected in the liver of experimentally infected hamsters induced a lesser intense granulomatous reaction than that induced by living bilharzial ova. This pathological reaction may be induced by immune complex formation.

Influence of the neonatal (clomiphene) inhibition of estradiol imprinting on gonadal development and gonadal and thyroid hormone production later in life.

Treatment of newborn rats with clomiphene citrate during the first 5 days of life gave rise to a marked decrease in body mass and to a still greater decrease in gonadal mass. A decrease was also observed in the testicular diameter of the males. The females showed a 43% increase in their estradiol levels over the control and an increase in the sensitivity to gonadotropins. Thyroxine level, which was also determined in view of the known gonadotropin-thyrotropin overlap, showed no change 6 weeks after pretreatment with clomiphene, while the thyroid gland responded to gonadotropin in the same manner as to thyrotropin.