Cartilage thickness in the hip joint measured by MRI and stereology – a methodological study

AbstractObjectiveThe purpose of this study was to develop a precise and efficient method for estimating the thickness of the articular cartilage in the hip joint and hence three different stereologic methods were tested based on Magnetic Resonance Imaging.DesignTwenty two females and four males with hip dysplasia underwent MRI. The thickness of the femoral and acetabular cartilage was estimated.ResultsThe results for all three methods showed that the observed total variance on cartilage thickness is small. The mean thickness of the acetabular cartilage measured by the three different methods ranged between 1.15mm and 1.46mm. The mean thickness for the femoral cartilage measured by the three different methods ranged between 1.18mm and 1.78mm. The measurements took 15–20min per hip to carry out.ConclusionMethods 1 and 3 are as precise but we favour method 3 because the measurements are done on images obtained through the center of the femoral head which means that the cartilage surface is intersected perpendicular and partial volume effect avoided. We suggest that this method can be advantageous for assessing the progression of osteoarthritis in dysplastic hips after periacetabular osteotomy

Chronic hypoxemia in late gestation decreases cardiomyocyte number but does not change expression of hypoxia-responsive genes

BACKGROUND: Placental insufficiency is the leading cause of intrauterine growth restriction in the developed world and results in chronic hypoxemia in the fetus. Oxygen is essential for fetal heart development, but a hypoxemic environment in utero can permanently alter development of cardiomyocytes. The present study aimed to investigate the effect of placental restriction and chronic hypoxemia on total number of cardiomyocytes, cardiomyocyte apoptosis, total length of coronary capillaries, and expression of genes regulated by hypoxia. METHODS AND RESULTS: We induced experimental placental restriction from conception, which resulted in fetal growth restriction and chronic hypoxemia. Fetal hearts in the placental restriction group had fewer cardiomyocytes, but interestingly, there was no difference in the percentage of apoptotic cardiomyocytes; the abundance of the transcription factor that mediates hypoxia-induced apoptosis, p53; or expression of apoptotic genes Bax and Bcl2. Likewise, there was no difference in the abundance of autophagy regulator beclin 1 or expression of autophagic genes BECN1, BNIP3, LAMP1, and MAP1LC3B. Furthermore, fetuses exposed to normoxemia (control) or chronic hypoxemia (placental restriction) had similar mRNA expression of a suite of hypoxia-inducible factor target genes, which are essential for angiogenesis (VEGF, Flt1, Ang1, Ang2, and Tie2), vasodilation (iNOS and Adm), and glycolysis (GLUT1 and GLUT3). In addition, there was no change in the expression of PKC-ε, a cardioprotective gene with transcription regulated by hypoxia in a manner independent of hypoxia-inducible factors. There was an increased capillary length density but no difference in the total length of capillaries in the hearts of the chronically hypoxemic fetuses. CONCLUSION: The lack of upregulation of hypoxia target genes in response to chronic hypoxemia in the fetal heart in late gestation may be due to a decrease in the number of cardiomyocytes (decreased oxygen demand) and the maintenance of the total length of capillaries. Consequently, these adaptive responses in the fetal heart may maintain a normal oxygen tension within the cardiomyocyte of the chronically hypoxemic fetus in late gestation.Kimberley J. Botting, I. Caroline McMillen, Heather Forbes, Jens R. Nyengaard, Janna L. Morriso

Time-dependent changes induced by acute stress in function and architecture of excitatory synapses in prefrontal and frontal cortex

Stressful life events impact on brain and bodily function and represent major risk factors for stress-related neuropsychiatric disorders. The response to stressful events can promote adaptive plasticity and improved cognition, when the physiological stress response is efficiently activated and inactivated in due time, or maladaptive and harmful effects, when the response is overused or dysregulated. In turn, the outcome of a maladaptive stress response can be associated with the triggering of brain, systemic and metabolic disorders. Chronic stress has been shown to induce reduction of density of synapses and dendrites in prefrontal and frontal cortex (PFC/FC), with concomitant impairments in neuronal activity and cognitive functions. Instead, the early and rapid effects of acute stress on synaptic function and plasticity are often opposite, with enhancement of glutamate release/transmission, increased number of spines and synapses, enhancement of synaptic strength. However, the delayed effects of acute stress have not been investigated, although this could give crucial information on the time- dependent changes in the brain stress response. We have previously characterized the synaptic effects of acute footshock (FS)-stress, which induces enhancement of glutamate release/transmission in PFC/FC, due to the increase of the readily releasable pool (RRP), in turn mediated by rapid non-genomic corticosterone action at synapses (Mol. Psy., 19:433-443, 2014). Here we have analyzed the effects of acute FS-stress in the PFC/FC of rats at different times after completion of the stress protocol. We found that acute stress induced early and sustained increase of RRP over time in excitatory perforated synapses, while the number of non-perforated and axo-spinous synapses was increased (without changes in vesicle pools). The total number of synaptic spines was increased up to 24 h, while apical dendrites showed decreased density 2 weeks after acute stress (with no significant changes at earlier times). In behavioral tests for working memory, FS-stress improved performance 2 h after stress and impaired it after 24 h. Changes in glutamate release, RRP, number of synapses and spines are blocked or attenuated by prior chronic treatment with the antidepressant desipramine. The different glutamatergic modifications in functional and morphological plasticity suggest a bi-phasic process, during which the stress response in PFC/FC may turn from early increased excitatory activation into its opposite. The identification of these points and the players involved in the switch are crucial for the understanding of the dynamics of stress-related pathology

Low density of CD3+, CD4+ and CD8+ cells is associated with increased risk of relapse in squamous cell cervical cancer

The purpose of this study was to investigate the prognostic value of the primary in situ cellular immune response in cervical squamous cell carcinoma. A study of 102 women treated for stage IB and IIA disease, between 1990 and 2000, was performed. Paraffin-embedded cervical tissue processed at the time of diagnosis was immunostained for CD3+ (T cells), CD4+ (T helper/regulatory T cells) and CD8+ (cytotoxic T cells) cells. Immune cell profile densities were estimated using stereology. Both intra- and peritumoural cell densities were estimated. Using Cox's proportional hazards regression modelling we found an increase in cell density to decrease the risk of relapse for all three cell types. The density of peritumoural CD3+ cells seems to have the strongest potential for predicting relapse. An increase in CD3+ cell density from 795 to 2043 cells per mm2 (25–75 percentile) reduced the hazard ratio to 0.27

Increased blood pressure in adult offspring of families with Balkan Endemic Nephropathy: a prospective study

BACKGROUND: Previous studies have linked smaller kidney dimensions to increased blood pressure. However, patients with Balkan Endemic Nephropathy (BEN), whose kidneys shrink during the course of the disease, do not manifest increased blood pressure. The authors evaluated the relationship between kidney cortex width, kidney length, and blood pressure in the offspring of BEN patients and controls. METHODS: 102 offspring of BEN patients and 99 control offspring of non-BEN hospital patients in the Vratza District, Bulgaria, were enrolled in a prospective study and examined twice (2003/04 and 2004/05). Kidney dimensions were determined using ultrasound, blood pressure was measured, and medical information was collected. The parental disease of BEN was categorized into three groups: mother, father, or both parents. Repeated measurements were analyzed with mixed regression models. RESULTS: In all participants, a decrease in minimal kidney cortex width of 1 mm was related to an increase in systolic blood pressure of 1.4 mm Hg (p = 0.005). There was no association between kidney length and blood pressure. A maternal history of BEN was associated with an increase in systolic blood pressure of 6.7 mm Hg (p = 0.03); paternal BEN, +3.2 mm Hg (p = 0.35); or both parents affected, +9.9 mm Hg (p = 0.002). There was a similar relation of kidney cortex width and parental history of BEN with pulse pressure; however, no association with diastolic blood pressure was found. CONCLUSION: In BEN and control offspring, a smaller kidney cortex width predisposed to higher blood pressure. Unexpectedly, a maternal history of BEN was associated with average increased systolic blood pressure in offspring

Renal histomorphology in dogs with pyometra and control dogs, and long term clinical outcome with respect to signs of kidney disease

<p>Abstract</p> <p>Background</p> <p>Age-related changes in renal histomorphology are described, while the presence of glomerulonephritis in dogs with pyometra is controversial in current literature.</p> <p>Methods</p> <p>Dogs with pyometra were examined retrospectively for evidence of secondary renal damage and persisting renal disease through two retrospective studies. In Study 1, light microscopic lesions of renal tissue were graded and compared in nineteen dogs with pyometra and thirteen age-matched control bitches. In Study 2, forty-one owners of dogs with pyometra were interviewed approximately 8 years after surgery for evidence ofclinical signs of renal failure in order to document causes of death/euthanasia.</p> <p>Results</p> <p>Interstitial inflammation and tubular atrophy were more pronounced in dogs with pyometra than in the control animals. Glomerular lesions classified as glomerular sclerosis were present in both groups. No unequivocal light microscopic features of glomerulonephritis were observed in bitches in any of the groups.</p> <p>Two bitches severely proteinuric at the time of surgery had developed end stage renal disease within 3 years. In five of the bitches polyuria persisted after surgery. Most bitches did not show signs of kidney disease at the time of death/euthanasia.</p> <p>Conclusion</p> <p>Tubulointerstitial inflammation was observed, but glomerular damage beyond age-related changes could not be demonstrated by light microscopy in the dogs with pyometra. However, severe proteinuria after surgery may predispose to development of renal failure.</p

A genetically modified minipig model for Alzheimer's disease with SORL1 haploinsufficiency

The established causal genes in Alzheimer’s disease (AD), APP, PSEN1, and PSEN2, are functionally characterized using biomarkers, capturing an in vivo profile reflecting the disease’s initial preclinical phase. Mutations in SORL1, encoding the endosome recycling receptor SORLA, are found in 2%–3% of individuals with early-onset AD, and SORL1 haploinsufficiency appears to be causal for AD. To test whether SORL1 can function as an AD causal gene, we use CRISPR-Cas9-based gene editing to develop a model of SORL1 haploinsufficiency in Göttingen minipigs, taking advantage of porcine models for biomarker investigations. SORL1 haploinsufficiency in young adult minipigs is found to phenocopy the preclinical in vivo profile of AD observed with APP, PSEN1, and PSEN2, resulting in elevated levels of β-amyloid (Aβ) and tau preceding amyloid plaque formation and neurodegeneration, as observed in humans. Our study provides functional support for the theory that SORL1 haploinsufficiency leads to endosome cytopathology with biofluid hallmarks of autosomal dominant AD

Developments in cell biology for quantitative immunoelectron microscopy based on thin sections: a review

Quantitative immunoelectron microscopy uses ultrathin sections and gold particle labelling to determine distributions of molecules across cell compartments. Here, we review a portfolio of new methods for comparing labelling distributions between different compartments in one study group (method 1) and between the same compartments in two or more groups (method 2). Specimen samples are selected unbiasedly and then observed and expected distributions of gold particles are estimated and compared by appropriate statistical procedures. The methods can be used to analyse gold label distributed between volume-occupying (organelle) and surface-occupying (membrane) compartments, but in method 1, membranes must be treated as organelles. With method 1, gold counts are combined with stereological estimators of compartment size to determine labelling density (LD). For volume-occupiers, LD can be expressed simply as golds per test point and, for surface-occupiers, as golds per test line intersection. Expected distributions are generated by randomly assigning gold particles to compartments and expressing observed/expected counts as a relative labelling index (RLI). Preferentially-labelled compartments are identified from their RLI values and by Chi-squared analysis of observed and expected distributions. For method 2, the raw gold particle counts distributed between compartments are simply compared across groups by contingency table and Chi-squared analysis. This identifies the main compartments responsible for the differences between group distributions. Finally, we discuss labelling efficiency (the number of gold particles per target molecule) and describe how it can be estimated for volume- or surface-occupiers by combining stereological data with biochemical determinations