Prostate cancer in men aged less than 50 years at diagnosis.

PURPOSE: Prostate cancer (CaP) in younger men (age ≤50 years) appears to present differently compared with older men. This study describes CaP characteristics and outcomes in Australian young men. METHODS: The South Australian Prostate Cancer Clinical Outcomes Collaborative database was used to identify men diagnosed with CaP 1998-2012. Men were stratified by age at diagnosis into groups ≤50, 50-70 and ≥70 years. Primary outcomes of cumulative biochemical recurrence (BCR) and cumulative prostate cancer-specific mortality (PCSM) were assessed at 5 and 10 years. RESULTS: In total, 7018 men were included. At time of diagnosis, 182 (2.6 %) were aged ≤50 years. Median follow-up exceeded 4 years. Younger men had a greater proportion of T stage <2 disease, lower median PSA and higher rates of Gleason score <7 (all p < 0.001). They were more likely to experience active surveillance (AS) (4.9, 3.1, 1.5 %) or radical prostatectomy (RP) (70, 55, 8 %) and less likely radiotherapy (13, 24, 29 %) as their principal modality (all p < 0.001). Although only 4.9 % underwent AS, 48 % of men ≤50 years were eligible for AS. Men ≤50 years had both the lowest unadjusted cumulative BCR and PCSM at 10 years. After multivariate analysis, BCR was not significantly different. Sample size limited multivariate analysis of PCSM. CONCLUSIONS: In our cohort, men ≤50 years with CaP had less aggressive clinical characteristics, but were more likely to undergo RP. They appear to experience lower unadjusted PCSM, but similar rates of adjusted BCR. Further studies are needed to assess whether AS is appropriately utilised in these men.

E-learning vs lecture: which is the best approach to surgical teaching?

AIM: Most medical teaching is still delivered by traditional face-to-face interaction. E-learning has the potential benefit of instilling deeper learning of topics by virtue of repeated and convenient access to content presented in a range of media. We aimed to evaluate objectively the benefit of educating medical students on a common surgical topic (haemorrhoids), through a website and podcast package vs a traditional lecture. METHOD: Baseline knowledge was established by a questionnaire given to two different groups of third-year medical students starting their first clinical attachment. Group A (n = 73) was given a lecture and group B (n = 75) was asked to use a website containing text and pictures augmented by a podcast. Students were reassessed using the same preintervention questionnaire, and satisfaction was acquired from details given in a feedback form. RESULTS: There was no difference in knowledge between the two groups at baseline. Both groups demonstrated significant gains in knowledge after intervention (P < 0.0001). Group B (Web/podcast) showed a significantly greater increase in knowledge (P < 0.05) than group A (lecture). Preintervention subjective assessment of knowledge rated by the students showed no difference between the groups. Both groups of students were equally satisfied with the educational method. CONCLUSION: E-learning supplemented with a podcast results in greater knowledge acquisition when compared with a traditional lecture, without a loss of satisfaction with teaching. Using augmented Web-based educational tools reduces demands on teaching time with no decrease in quality for selected parts of the curriculum.

Fiberoptic bronchoscopy in nonhospitalized patients.

Fiberoptic bronchoscopy is employed extensively in the examination of patients with pulmonary disease. As demonstrated in this large series, the procedure can be performed safely and efficiently on an outpatient basis, with the use of only topical anesthesia. Discomfort is minimal and readily accepted by most patients. Bronchial biopsies of suspected neoplastic lesions can be performed without difficulty. Outpatient bronchoscopy in an appropriate clinical setting can hasten the medical evaluation without compromising patient care.

Crystallochemical characteristics of the inorganic phase of human calcific aortic valvar stenosis.

To determine the elemental composition of the mineral deposits found in human calcific aortic valvar stenosis and to verify by x ray diffraction crystal class previously reported by electron diffraction 138 quantitative energy dispersive analyses and x ray diffraction of pooled specimens were performed on samples of aortic valves excised from 12 patients. The principal components of the calcified portions of the valves were 38.15% calcium and 17.68% phosphorus. The weight percentage calcium to phosphorus ratio was 2.16 and an elemental distribution related to the simplest ideal unit cell formulation Ca10(PO4)6(OH)2 was determined as: (Ca0.952Mg0.044Na0.097K0.005)10 (P0.952S0.025C0.023)6O23.96(Cl0.0184O0. 982H)2. The hexagonal unit cell was found to have an a axis of 9.424 A(0.9424 nm); c axis of 6.787 A(0.6787 nm); volume of 522 A3(0.522 nm3); and density of 3.2 g.cm-3. On the bases of the composition, the axial dimensions, and the density the material was classified as the dahllite form of apatite. There was no evidence of crystalline cholesterol. Variations in elemental composition showed no significant correlation with age or sex of the patient; statistically, neither the cusp involved nor the location on the leaflet affected the mineral composition.

Physical chemical evidence of structural weakness in coronary arterial calcification.

Physical-chemical properties of calcified human coronary arteries were obtained through 857 electron probe microanalyses of samples of endarterectomy tissue from 12 patients and by X-ray and electron diffraction from separate samples. The calcific phase exhibited a weight percentage calcium to phosphorus ratio of 2.5 (SEM 0.15), a density of 2.95 (0.02) g.cm-3, and a crystalline unit cell volume of 530 (0.5) A3, or 53 (0.05) nm3. The phase has been characterised as a phosphorus deficient dahllite variant of apatite in which the crystal structure has been weakened by the incorporation of water molecules and tetrahedral hydroxyl groups in isomorphic substitution conforming with the McConnell-type defect. Isomorphic substitution within the apatitic cell appears sufficient to suggest that the crystal structure of the mineral deposit is weakened to the extent that means might be sought to attack the early formation of the pathological deposition.

Follicular growth and atresia during the last half of the luteal phase of the guinea pig estrous cycle: relation to serum progesterone and estradiol levels and utero-ovarian blood flow.

The relationships among follicular growth, changes in serum progesterone (P) and estradiol (E) levels, and utero-ovarian blood flow (OBF) through the guinea pig ovary were examined during days 7-16 of the luteal phase (day 0 = estrus) of the estrous cycle and during the subsequent ovulatory (proestrous-estrous) period. Follicles were classified as either viable or atretic based on strict criteria and grouped according to diameter. No changes in follicular growth were observed between days 7-9 of the cycle when serum P levels were elevated and OBF was at low rates. Between days 13-15 when serum P levels were low and both OBF and serum E levels were rising, there was a dramatic increase in the number of viable follicles present in all follicle classes. As the percentage of viable follicles increased, the number of atretic follicles in each size population decreased. Peaks in the number of large follicles (300-450 microns), OBF, and serum E levels were observed during the subsequent ovulatory period. These data suggest that as luteal activity declines during the last half of the estrous cycle, follicular recruitment and growth are stimulated. The concomitant elevations in OBF suggest a supportive role for this parameter in follicular development. In turn, the subsequent elevation in serum E levels serves as an index of follicular maturation. These data suggest that the elevated P levels during the luteal phase of the estrous cycle may either directly or indirectly, through the regulation of gonadotropin secretion, regulate follicular growth in the guinea pig.

Cell attachment to, and contraction of, the retina in vitro.

PURPOSE: To examine the behavior of fibroblasts and retinal pigment epithelial cells after attachment to the retinal surface in vitro to elucidate the pathobiology of the early stages of epiretinal membrane formation. METHODS: Human retinal pigment epithelial (HRPE) cells and bovine Tenon's capsule fibroblasts (BTFs) were seeded onto the surface of bovine retinal explants maintained in organ culture. The changes induced in the underlying retina, including contraction, were assessed during a period of up to 10 days. Immunohistochemistry was used to assess proliferation of the seeded cells and to determine deposition of extracellular matrix. RESULTS: Explants of bovine neuroretina were maintained in organ culture, with good morphologic preservation of the inner limiting lamina and inner retinal layers, for 7 to 10 days. The HRPE cells and the BTFs attached to the retinal surface and exerted tractional forces, producing partial- and full-thickness retinal folding. Contraction commenced within 24 hours of attachment of the cells and continued for several days, with most of the contraction occurring within the next 48 to 72 hours. The HRPE cells and BTFs were found to be equally contractile. Deposition of cellular fibronectin (but not collagen type I) was demonstrated. CONCLUSIONS: The contractile cellular membranes generated in this organ culture system exhibit many of the morphologic and functional features of epiretinal membranes found in the early stages of proliferative vitreoretinopathy.

Expression of a single pair of desmosomal glycoproteins renders the corneal epithelium unique amongst stratified epithelia.

PURPOSE: To determine desmosomal glycoprotein isoform expression in bovine corneal, limbal, and conjunctival epithelium and desmosomal profile and distribution during corneal re-epithelialization. METHODS: Immunofluorescence (IF) for desmosomal components on cryostat sections of fresh epithelia was supported by immunoblot analysis of tissue lysates. Wounded corneas maintained in organ culture were examined by IF at times up to full re-epithelialization (96 hours). RESULT: Immunofluorescence for desmoplakin confirmed desmosome presence throughout all three epithelia. Plakoglobin was also ubiquitous. Of the desmosomal glycoproteins, desmocollin 2 (Dsc2) and desmoglein 2 (Dsg2) were expressed throughout, but Dsc3 and Dsg3 were confined to the limbus and conjunctiva, and Dscl and Dsgl were absent. Dsc2 and Dsg2 IFs were stronger in superficial layers, but Dsc3 and Dsg3 were stronger basally, fading suprabasally. Glycoprotein expression in cornea and conjunctiva was confirmed by immunoblot analysis. No change in glycoprotein expression occurred during re-epithelialization. CONCLUSIONS: Uniquely among stratified epithelia, cornea expresses only a single pair of desmosomal glycoproteins, Dsc2 and Dsg2. Expression of Dsc3 and Dsg3 in limbus and conjunctiva coincides with their association with cell proliferation in other epithelia, but corneal epithelial cells did not express Dsc3 or Dsg3 during re-epithelialization. Absence of Dscl and Dsgl correlates with lack of keratinization in ocular epithelia. These expression patterns may have significance for the specific properties and differentiation patterns of the epithelia. Presence of desmosomes throughout re-epithelialization raises the question of how migrating cells mutually re-position.

31P nuclear magnetic resonance (NMR) identification of sugar phosphates in isolated rat ovarian follicular granulosa cells and the effects of follicle-stimulating hormone.

Immature female rats (23-30 days old) were implanted subcutaneously with diethylstilbestrol (DES) in silastic capsules. After 48 h their ovaries were removed and the granulosa cells isolated (Foreman et al. (1984) Life Sci. 35, 1273-1279). The cells were incubated in Hepes balanced saline buffer with substrates with or without follicle-stimulating hormone (FSH). At the end of incubation perchloric acid extracts were made for 31P NMR spectroscopy. The resonances of fructose 1-phosphate, fructose 6-phosphate, glucose 1-phosphate, and ribose 5-phosphate were identified in the granulosa cell extracts. The relative intensities of fructose 6-phosphate to ribose 5-phosphate decreased after incubation with FSH in vitro. This suggests that FSH increases the activity of the pentose pathway within 1 h. Thus, FSH can acutely activate those metabolic pathways which provide nicotinamide-adenine dinucleotide phosphate (NADPH) to be used in steroid synthesis and cholesterol mobilization.

Efficacy of 67Ga-scintigraphy in predicting the diagnostic yield of transbronchial lung biopsy in pulmonary sarcoidosis.

Nineteen consecutive patients with clinically suspected sarcoidosis underwent 67Ga-scintigraphy prior to transbronchial lung biopsy (TBLB) to determine if 67Ga uptake in lung parenchyma would increase the diagnostic yield of the biopsy procedure. Biopsies were obtained from the areas showing parenchymal uptake on the 67Ga scan in 13 of the 19 patients. In the six patients not demonstrating uptake of 67Ga in the lung parenchyma, biopsies were obtained at random from the right lower lobe. There was no correlation between 67Ga uptake in hilar nodes or pulmonary parenchyma tissue and the diagnostic yield from TBLB. We conclude that 67Ga scanning is neither efficacious nor cost-effective in predicting the diagnostic yield of TBLB in sarcoidosis.

Diffusion of strontium from desensitizing agents into human dentin as determined by energy-dispersive analysis.

Strontium-containing desensitizing agents were applied to one set of tooth surfaces and were burnished onto another set of diametric surfaces. Concentration of strontium at dentin depth was determined by X-radiation dispersive analysis. Burnishing produced deeper strontium penetration. Fick's Second Law provided an estimate of relative effective time for equal strontium distributions by the two methods.

Optical method for articulator comparison and study of occlusal determinants.

A method of articulator comparison and a technique demonstrating the effects of three occlusal determinants were demonstrated and proved successful. The techniques used could be used in dental research circumstances and in dental schools as a means to study the determinants of occlusion.

Electron microscope comparison of freeze-substitution and conventional chemical fixation of undecalificied human dentin.

Within the stated limits of cell fine structure preservation and within the observed anatomical limits specified in the Results and Discussion sections, a freeze-substitution method using an ethylene glycol-Hank's solution eutectic mixture with a glutaraldehyde additive can be used to effectively prepare undecalcified human dentin for electron microscopy. The ultrastructual appearance of the odontoblast cell body and the odontoblastic process subjected to freeze-substitution differs from that seen with conventional chemical fixation. Artifacts produced by freeze-substitution differ in appearance and frequency of occurrence from those produced by glutaraldehyde-osmium tetroxide sequential double fixation. The cellular component of dentin shows greater structural preservation of protein when it is subjected to freeze-substitution that when it is prepared by conventional chemical fixation. The absence of ice crystal defects in the odontoblastic process in calcified dentin and the presence of ice crystal defects in the odontoblast cell body suggest that intracellular water in the odontoblastic process in the calcified dentin may exist in a more highly structured state than intracellular water in the odontoblast cell body. If intracellular water exists in a more highly structured state in the odontoblastic process of the calcified dentin than in the cell body, the ratio of protein molecules to cytoplasmic volume may be greater in the odontoblastic process than in the cell body. After glutaraldehyde-osmium tetroxide sequential double fixation, the use of graded alcohol dehydration obtained cell fine structure preservation and artifact control superior to that obtained by use of ethylene glycol cryodehydration. Further refinements of the freeze-substitution technique, as it applies to the preparation of undecalcified human dentin, are necessary to increase the amount of cellular preservation, to decrease the number of ice crystal artifacts, and to improve the overall quality of cell fine structure preservation.

VEGF localisation in diabetic retinopathy.

AIM: To determine the staining pattern of vascular endothelial growth factor (VEGF) at different stages of diabetic retinopathy (including post-laser photocoagulation) and to compare staining in excised fibrovascular and fibrocellular (non-diabetic) preretinal membranes. METHODS: Immunohistochemical localisation of VEGF, using antibodies raised against VEGF165 and VEGF121,165,189, was carried out on specimens of normal human retina (n = 15), diabetic retinas ((a) with no overt retinopathy (n = 19), (b) with intraretinal vascular abnormalities but no proliferative retinopathy (n = 6), (c) with active proliferative retinopathy (n = 6), (d) with no residual proliferative retinopathy after photocoagulation therapy (n = 15)), excised diabetic fibrovascular membranes (n = 19), and non-diabetic fibrocellular membranes (n = 7). The degree and pattern of immunostaining was recorded. RESULTS: In general, VEGF was absent from the majority of normal retinas. VEGF staining was apparent in most diabetic tissues but the staining pattern was dependent on both the specificity of the antibody used and the category of tissue. Staining with the VEGF165 antibody was generally confined to endothelial cells adn perivascular regions while the VEGF121,165,189 antibody was also associated with extravascular components of the inner retina. Intensity of immunostaining of diabetic eyes was dependent on the severity of retinopathy being least in diabetics with no overt retinopathy and greatest in retinas with proliferative retinopathy. Interestingly, the intensity of immunostaining in diabetic retinas which had undergone laser surgery for proliferative retinopathy was reduced to basal levels. Moderate to intense immunostaining was observed in all fibrovascular and fibrocellular membranes examined. CONCLUSIONS: This study supports a circumstantial role for VEGF in the pathogenesis of both the preclinical and proliferative stages of diabetic retinopathy.

Immunolocalisation of the VEGF receptors FLT-1, KDR, and FLT-4 in diabetic retinopathy.

AIM: To determine the spatial and temporal changes in the staining pattern of the VEGF receptors FLT-1, KDR, and the putative receptor FLT-4 during the pathogenesis of diabetic retinopathy. METHODS: Immunohistochemical localisation of VEGF receptors, using antibodies against FLT-1, FLT-4, and KDR, was carried out on specimens of normal human retina (n = 10), diabetic retinas (a) with no overt retinopathy (n = 12), (b) with intraretinal vascular abnormalities but no proliferative retinopathy (n = 5), (c) with active proliferative retinopathy (n = 6), and (d) with no residual proliferative retinopathy after scatter photocoagulation therapy (n = 14), and surgically excised diabetic fibrovascular membranes (n = 11). The degree and pattern of immunostaining was recorded. RESULTS: FLT-1 staining was apparent in the retinas from both non-diabetic and diabetic retinas; weak to moderate staining was generally confined to the inner nuclear layer, the ganglion cell layer, and the retinal vessels during all stages of the disease process. Staining of the retinal vessels was raised in diabetic tissue compared with non-diabetic tissue. The preretinal vessels of the diabetic subjects stained moderately to intensely for FLT-1. In contrast with FLT-1 staining minimal immunostaining for KDR was demonstrated in the non-diabetic eyes and the unlasered eyes; however, weak staining for KDR was observed in the inner nuclear layer and the ganglion cell layer of the unlasered eyes with diabetic changes. In those retinas with preretinal neovascularisation KDR immunoreactivity was moderate to intense in the intra- and preretinal vessels. However, in the excised membranes, where the vessels may have been in a quiescent state, the levels of KDR were weak to moderate. After apparently successful laser treatment KDR staining was reduced in the intraretinal vessels. Minimal FLT-4 staining was observed throughout normal eyes while weak to moderate FLT-4 staining was generally confined to the inner nuclear layer and the ganglion cell layer of the unlasered diabetic eyes. Weak to moderate levels of FLT-4 staining were observed in the intraretinal vessels except after apparently successful laser treatment where reduced levels of staining were observed. Weak to moderate staining was observed in the preretinal vessels. CONCLUSIONS: This study supports a role for FLT-1, KDR, and possibly FLT-4 in the pathogenesis of diabetic retinopathy; however, their specific roles in the progression of the disease may differ.

Three dimensional analysis of the retinal vasculature using immunofluorescent staining and confocal laser scanning microscopy.

AIM: To undertake a qualitative and quantitative analysis in three dimensions of the human retinal vasculature. METHOD: Fixed and excised whole retinas were permeabilised and subjected to immunofluorescent staining for blood vessel components followed by confocal laser scanning microscopy. Single projection and stereoimages were constructed using computer software. XZ sections through the retina were constructed and the vasculature analysed using appropriate software. RESULTS: Immunofluorescent staining with no discontinuities was present in vessels of all sizes, the confocal images of the capillary network being free of out of focus blur at all depths. Quantitative analysis of XZ sections confirmed the qualitative impression of sharp delineation of the deep retinal capillary plexus, an absence of laminar arrangement of capillaries within the inner retina, and a truncated cone of capillaries around the foveal avascular zone (FAZ) wherein the superficial capillaries approached the FAZ more closely than those in the deeper retina. CONCLUSION: Immunofluorescent staining of the retina and confocal laser scanning microscopy were shown to be useful in analysing accurate three dimensional reconstructions of the normal retinal vasculature without affecting overall tissue architecture.

Intravitreal growth factors in proliferative diabetic retinopathy: correlation with neovascular activity and glycaemic management.

AIM: Many growth factors are implicated in proliferative diabetic retinopathy (PDR). It was decided to test the hypothesis that no one factor is predominant but that a regular profile of levels of different growth factors might be operating, and that the profile might differ according to whether or not insulin therapy was part of the patient's glycaemic management. The levels of several growth factors in vitrectomy samples were therefore determined from diabetic patients with tractional, non-haemorrhagic sequelae of PDR and these levels were correlated with (a) each other (growth factor profile), (b) neovascular activity, and (c) the method of glycaemic management (insulin treated (IT) or non-insulin treated (NIT)). METHODS: 72 samples of vitreous were obtained from either diabetic patients with PDR (n = 51) or non-diabetic (control) patients (n = 21). Levels of bFGF, IGF-I, EGF, and insulin were determined by radioimmunoassay; levels of TGF-beta 2 by ELISA; and levels of IGF-I binding protein by western ligand blotting. The data were analysed using appropriate statistics. RESULTS: There was no regular growth factor profile. bFGF levels were significantly greater in vitreous from NIT patients compared with IT patients and controls. The highest levels of bFGF were found in NIT patients with actively vascularised membranes. TGF-beta 2 levels were significantly greater in vitreous from IT patients compared with NIT patients and controls The highest levels of TGF-beta 2 were found in IT patients with actively vascularised membranes. IGF-I levels were significantly greater in diabetics (irrespective of insulin treatment) than non-diabetics and the highest levels of IGF-I were found in IT patients with actively vascularised membranes. A 34 kDa IGFBP was the predominant IGFBP identified in vitreous and was found to be elevated in diabetics patients. CONCLUSION: In PDR there is a correlation between intravitreal growth factor levels and both disease state (whether active or fibrotic) and method of glycaemic management.

The effects of gonadotropins on metabolism of isolated rat granulosa cells.

FSH in vitro, but not LH, increased the O2 uptake of isolated granulosa cells from 23 day old rats previously treated with DES or with DES and FSH. Dose response studies showed that the cells were most sensitive to FSH when the cellular binding of FSH was highest. LH increased the O2 uptake of granulosa cells of untreated 30 day old rats. DES treatment inhibited the LH induced rise in O2 uptake when the rats were implanted with DES capsules unless FSH was injected to induce LH receptors. Addition of dbcAMP in vitro increased O2 uptake of granulosa cells from 30 day old rats at concentrations 10X lower than those required to stimulate O2 uptake in cells from 23 day old rats treated with DES alone.

Histologic evaluation of periodontal attachment apparatus following the insertion of a biodegradable copolymer barrier in humans.

The purpose of this study was to histologically evaluate the effectiveness of polylactide:polyglycolide 50:50 (DL-PLGA) as a barrier to prevent epithelial migration and to promote new connective tissue attachment. Mucoperiosteal flaps were performed on 17 human teeth. DL-PLGA membrane was placed over the roots and alveolar bone. DL-PLGA was not placed over control teeth. Ninety days following surgery, block sections were obtained and processed for histologic evaluation. All specimens revealed a mean length of epithelial attachment within physiologic limits and recession of the gingival margin. There was little or no reattachment of the connective tissue. It is concluded that this copolymer barrier did not prevent epithelial migration nor enhance connective tissue attachment to human roots with severe horizontal bone loss and active periodontal disease.