Infrapatellar Fat Pad Gene Expression and Protein Production in Patients with and without Osteoarthritis.

Osteoarthritis (OA) is one of the most common joint disorders. Evidence suggests that the infrapatellar fat pad (IFP) is directly involved in OA pathology. However, a comparison between OA versus non-OA IFP is still missing. Thus, the aim of this study was to compare IFP molecular, adipocytes and extracellular matrix characteristics of patients affected by OA, and patients undergoing anterior cruciate ligament (ACL) reconstruction. We hypothesized that not only inflammation but also changes in adipocytes and extracellular matrix (ECM) composition might be involved in OA pathogenesis. Fifty-three patients were enrolled. IFP biopsies were obtained, evaluating: (a) lymphocytic infiltration and vascularization; (b) adipocytes area and number; (c) adipo-cytokines and extracellular matrix gene expression levels; (d) IL-6 and VEGF protein production; (e) collagen fibers distribution. OA IFP was more inflamed and vascularized compared to ACL IFP. OA IFP adipocytes were larger and numerically lower (1.3-fold) than ACL IFP adipocytes. An increase of gene expression of typical white adipose tissue genes was observed in OA compared to ACL IFP. Collagen-types distribution was different in the OA IFP group compared to controls, possibly explaining the change of the biomechanical characteristics found in OA IFP. Statistical linear models revealed that the adipocyte area correlated with BMI in the OA group. In conclusion, inflammation and fibrotic changes of OA IFP could represent novel therapeutic targets to counteract OA.

Infrapatellar fat pad features in osteoarthritis: a histopathological and molecular study.

Objective: The infrapatellar fat pad (IFP) is considered a local producer of adipocytokines, suggesting a potential role in OA. The objective of this study was to evaluate the histopathological and molecular characteristics of OA IFPs compared with controls. Methods: The histopathological characteristics of IFPs were evaluated in patients undergoing total knee replacements and in control patients (without OA), considering the following parameters: presence of inflammatory cells, vascularization, adipose lobules dimension and thickness of the interlobular septa. Immunohistochemistry was performed to evaluate VEGF, monocyte chemotactic protein 1 (MCP-1) and IL-6 proteins. Quantitative real time PCR was performed to evaluate the expression levels of adipocytokines in the OA IFPs. Results: OA IFPs showed an increase in inflammatory infiltration, vascularization and thickness of the interlobular septa compared with controls. VEGF, MCP-1 and IL-6 proteins were higher in OA IFPs compared with in controls. Inflammatory infiltration, hyperplasia, vascularization and fibrosis were increased in OA IFP synovial membranes compared with in those of controls. VEGF protein levels were associated with an increased number of vessels in the OA IFPs, while MCP-1 and IL-6 protein levels were associated with higher grades of inflammatory infiltration. Leptin levels were positively correlated with adiponectin and MCP-1expression, while adiponectin positively correlated with peroxisome proliferative activated receptor gamma, MCP-1 and IFP vascularity. MCP-1 showed a positive correlation with peroxisome proliferative activated receptor gamma. IFP lobules dimensions were positively correlated with IL-6 expression and negatively with thickness of interlobular septa. VEGF mRNA levels were positively correlated with increased synovial vascularity. Conclusions: OA IFPs and synovial membranes are more inflamed, vascularized and fibrous compared with those of control patients (without OA).

Fractal analysis of alveolarization in hyperoxia-induced rat models of bronchopulmonary dysplasia.

No papers are available about potentiality of fractal analysis in quantitative assessment of alveolarization in bronchopulmonary dysplasia (BPD). Thus, we here performed a comparative analysis between fractal [fractal dimension (D) and lacunarity] and stereological [mean linear intercept (Lm), total volume of alveolar air spaces, total number of alveoli, mean alveolar volume, total volume and surface area of alveolar septa, and mean alveolar septal thickness] parameters in experimental hyperoxia-induced models of BPD. At birth, rats were distributed between the following groups: 1) rats raised in ambient air for 2 wk; 2) rats exposed to 60% oxygen for 2 wk; 3) rats raised in normoxia for 6 wk; and 4) rats exposed to 60% hyperoxia for 2 wk and to room air for further 4 wk. Normoxic 6-wk rats showed increased D and decreased lacunarity with respect to normoxic 2-wk rats, together with changes in all stereological parameters except for mean alveolar volume. Hyperoxia-exposed 2-wk rats showed significant changes only in total number of alveoli, mean alveolar volume, and lacunarity with respect to equal-in-age normoxic rats. In the comparison between 6-wk rats, the hyperoxia-exposed group showed decreased D and increased lacunarity, together with changes in all stereological parameters except for septal thickness. Analysis of receiver operating characteristic curves showed a comparable discriminatory power of D, lacunarity, and total number of alveoli; Lm and mean alveolar volume were less discriminative. D and lacunarity did not show significant changes when different segmentation thresholds were applied, suggesting that the fractal approach may be fit to automatic image analysis.

The Infrapatellar Adipose Body: A Histotopographic Study.

The infrapatellar fat pad (IFP) can be regarded as a peculiar form of fibro-adipose tissue localized close to the synovial membrane and articular cartilage. The aims of the present study were to analyze the microscopic anatomy of the IFP through histological and ultrastructural methods, comparing it with that of the subcutaneous tissue of the abdomen and of the knee. Ten specimens of IFP were sampled from bodies of the Donation Program of the University of Padua without a history of osteoarthritis. The IFP consisted of white adipose tissue, of lobular type, with lobules delimited by thin connective septa. The IFP lobule areas were smaller (p < 0.05) and the interlobular septa were thicker (p > 0.05) than those of subcutaneous tissues of the abdomen, whereas the IFP lobule areas were larger (p < 0.05) and the interlobular septa were thinner than those of the subcutaneous tissue of the knee (p < 0.05). The IFP adipocytes present a mean area of 3,708 ± 976 µm2 with a large intercellular space, whereas the mean area of the abdominal tissues was greater (6,082 ± 628 µm2; p < 0.05). At scanning electron microscopy the IFP adipocytes were covered by thick fibrillary sheaths, creating a basket around the adipocytes. The structural characteristics of the IFP (lobular aspect of the adipose tissue, thickness of the septa with scarce elastic fibers) could act as a plastic portion aimed at the absorption of pressure variation during knee articular activity. The extensive distribution of nerves suggests a possible role of the IFP as a mechanoreceptor, corresponding to a tridimensional connective mesh working in the proprioceptive regulation of the activity of the knee joint.

Tissue Dynamics of the Carotid Body Under Chronic Hypoxia: A Computational Study.

The carotid body (CB) increases in volume in response to chronic continuous hypoxia and the mechanisms underlying this adaptive response are not fully elucidated. It has been proposed that chronic hypoxia could lead to the generation of a sub-population of type II cells representing precursors, which, in turn, can give rise to mature type I cells. To test whether this process could explain not only the observed changes in cell number, but also the micro-anatomical pattern of tissue rearrangement, a mathematical modeling approach was devised to simulate the hypothetical sequence of cellular events occurring within the CB during chronic hypoxia. The modeling strategy involved two steps. In a first step a "population level" modeling approach was followed, in order to estimate, by comparing the model results with the available experimental data, "macroscopic" features of the cell system, such as cell population expansion rates and differentiation rates. In the second step, these results represented key parameters to build a "cell-centered" model simulating the self-organization of a system of CB cells under a chronic hypoxic stimulus and including cell adhesion, cytoskeletal rearrangement, cell proliferation, differentiation, and apoptosis. The cell patterns generated by the model showed consistency (from both a qualitative and quantitative point of view) with the observations performed on real tissue samples obtained from rats exposed to 16 days hypoxia, indicating that the hypothesized sequence of cellular events was adequate to explain not only changes in cell number, but also the tissue architecture acquired by CB following a chronic hypoxic stimulus.

The cubital tunnel: a radiologic and histotopographic study.

Entrapment of the ulnar nerve at the elbow is the second most common compression neuropathy in the upper limb. The present study evaluates the anatomy of the cubital tunnel. Eighteen upper limbs were analysed in unembalmed cadavers using ultrasound examination in all cases, dissection in nine cases, and microscopic study in nine cases. In all cases, thickening of the fascia at the level of the tunnel was found at dissection. From the microscopic point of view, the ulnar nerve is a multifascicular trunk (mean area of 6.0 ± 1.5 mm(2) ). The roof of the cubital tunnel showed the presence of superimposed layers, corresponding to fascial, tendineous and muscular layers, giving rise to a tri-laminar structure (mean thickness 523 ± 235 µm). This multilayered tissue was hyperechoic (mean thickness 0.9 ± 0.3 mm) on ultrasound imaging. The roof of the cubital tunnel is elastic, formed by a myofascial trilaminar retinaculum. The pathological fusion of these three layers reduces gliding of the ulnar nerve during movements of the elbow joint. This may play a role in producing the symptoms typical of cubital tunnel syndrome. Independent from the surgical technique, decompression should span the ulnar nerve from the triceps brachii muscle to the flexor carpi ulnaris fascia.

Hyperbaric air exposure at 2.5 ATA does not affect respiratory mechanics and lung histology in the rat.

BACKGROUND: We previously demonstrated that the exposure to hyperbaric hyperoxia increased respiratory system elastance and both the "ohmic" and viscoelastic components of inspiratory resistances, probably because of increased oxygen tension toxic effects. We presently investigated the possible consequences of a single exposure to 2.5-atmospheres absolute air (hyperbarism) lasting 90 min. METHODS: We used the end-inflation occlusion method on anesthetized rats after about 15 min from previous exposure to hyperbarism. The method allows the measurements of respiratory system elastance and of the ohmic and viscoelastic components of airway resistance, which respectively depend on the Newtonian pressure dissipation due to the ohmic airway resistance to airflow and on the viscoelastic pressure dissipation caused by respiratory system tissue stress relaxation. The expressions of inducible NO synthase (iNOS) and endothelial NO synthase (eNOS) in the lung's tissues were also investigated, together with the histological characteristics of lung tissue. Data were compared with those obtained in control animals and in previously studied animals exposed to hyperoxic hyperbarism. RESULTS: Unlike with hyperoxic hyperbarism, hyperbarism per se did not change significantly the parameters of respiratory mechanics in the control animals (respiratory system elastance and ohmic and viscoelastic resistances were 2.01 ± 0.17 vs. 1.74 ± 0.08 cm H(2)O/ml, and 0.13 ± 0.02 vs. 0.13 ± 0.03 and 0.425 ± 0.04 vs. 0.33 ± 0.03 cm H(2)O/ml s(-1) in control vs. experimental animals, respectively, none significantly different), nor did it induce evident effects on lung histology. An increment of both iNOS and eNOS expressions was documented instead (0.50 ± 0.05 vs. 0.75 ± 0.07 and 1.04 ± 0.1 and 1.4 ± 0.15, respectively). CONCLUSION: Our results indicate that, at variance with hyperoxic hyperbarism, the acute exposure to only hyperbarism does not affect either the elastic or the resistive respiratory system properties, or lung histology.

Spexin is expressed in the carotid body and is upregulated by postnatal hyperoxia exposure.

Spexin is a recently identified peptide which is expressed in many different endocrine and nervous tissues. Due to the absence of data regarding spexin expression in the carotid body, the first aim of the present study was to investigate, through immunohistochemistry and Real-Time PCR, the expression and distribution of spexin in the rat and human carotid body. Moreover, the carotid body is known to undergo various structural and functional modifications in response to hyperoxic stimuli during the first postnatal period. Thus, we also evaluated if hyperoxia during the first postnatal weeks may produce changes in the spexin expression. Materials consisted of carotid bodies obtained at autopsy from five human adult subjects and sampled from 10 six-weeks old Sprague-Dawley rats. Five rats were maintained in normoxia for the first six postnatal weeks; five rats were exposed to 60% hyperoxia for 2 weeks and then maintained in normoxia for other 4 weeks. Diffuse anti-spexin immunoreactivity was found in type I cells of both humans and rats. No spexin immunoreactivity was visible in the type II cells. Hyperoxia exposure during the first 2 weeks of postnatal life caused a reduction of volume in the carotid body still apparent after 4 weeks of normoxia. Using real-time PCR, spexin expression was 6-7 times higher in hyperoxia-exposed rats than in normoxia-exposed ones. The expression of spexin in type I cells suggests a possible modulator role in peripheral chemoreception. Moreover, the ascertained role of spexin in the regulation of cell proliferation in other tissues (e.g., adrenal gland cortex) suggests a possible role of spexin also in the hyperoxia-induced plasticity of the carotid body.

High-quality Digital 3D Reconstruction of Microscopic Findings in Forensic Pathology: The Terminal Pathway of a Heart Stab Wound.

High-quality digital three-dimensional (3D) reconstructions of microscopic findings have been used in anatomical and histopathologic research, but their use in forensic pathology may also be of interest. This paper presents an application of these methods to better characterize the pathway of a stab wound of the anterior surface of the heart in a case of suicide. A portion of the heart wall including the stab wound was serially sectioned for microscopic analysis along the full extent of the wound. Histologic sections were digitally acquired, and a 3D reconstruction was created with ImageJ software for 3D computer graphics. This showed a full-thickness wound path extending to the endocardial surface of the left ventricle, curvilinear in appearance. After correction for shrinkage, 3D reconstruction allowed estimation of the dimensions of the myocardial injury and comparison of the appearance of the wound with the suspected knife used. The curvilinear appearance was considered to reflect injury during myocardial contraction. Complete microscopic sectioning and 3D reconstruction may allow virtual sectioning through various orientations and also provide useful forensic information for selected injuries.

Varicocele is associated with an increase of connective tissue of the pampiniform plexus vein wall.

OBJECTIVE: The purpose of this study is to describe the normal structure of spermatic vein wall and the morphological alterations in patients with grades 1-3 varicocele. METHODS: We dissected and analyzed a 2-3 cm tract of a pampiniform plexus vein of 30 patients undergoing antegrade scrotal sclerotherapy for left varicocele and of 13 consecutive patients undergoing radical orchidectomy for left testis cancer. The histological and morphological examination was performed with hematoxylin-eosin and Azan-Mallory staining through a computer image analysis approach. Vasa vasorum number was evaluated through the anti-CD31 monoclonal antibody. RESULTS: In normal veins, the tunica media had a circular smooth muscle layer, the tunica adventitia a longitudinal one; oblique fibers connected the longitudinal with the circular layer. In grade 1 varicocele, we found an increase in the connective tissue of the adventitia (39 vs. 1%, P < 0.001) and of the tunica media (13 vs. 0.5%, P = 0.001) and a decrease of oblique fibers. In grade 2 varicocele, the adventitia and media showed greater connective tissue (40 and 15%, P < 0.001). In grade 3 varicocele, adventitia and media showed a further increase in connective tissue (52 and 28%, respectively, <0.001). No oblique fibers were found in these veins. Vasa vasorum were decreased in number in patients with grades I and II varicocele, and were absent in patients with grade III varicocele. CONCLUSION: The wall of pampiniform plexus veins is a complex structure. Varicocele induces a decrease of vasa vasorum number, a fibrotic degeneration of the adventitia and media, and a decrease of the oblique running fibers.

Correction: Saccharomyces boulardii CNCM I-745 supplementation reduces gastrointestinal dysfunction in an animal model of IBS.

[This corrects the article DOI: 10.1371/journal.pone.0181863.].

Investigation of biomechanical response of Hoffa's fat pad and comparative characterization.

The infrapatellar adipose body (Hoffa's fat pad, IFP) is situated between the patellar tendon, the femoral condyle and the tibial plateau. The IFP consists of lobules of white adipose tissue delimited by thin connective septa. The actual structural functionality of the IFP is debated and should pertain to a cushioning role in the knee joint, providing to distribute and to damp mechanical stresses during articular activity. The present study is aimed to analyze the correlation between histological configuration and mechanical properties of the IFP, compared to other adipose tissues, partially differentiated by composition and conformation. Histological and ultrastructural methods were exploited to analyze the microscopic anatomies of IFP, knee (KSF) and abdominal (ASF) subcutaneous fat tissues. Numerical micro-models of the different tissues were developed by using histo-morphometric data, as the size of adipose lobules, the thickness of the septa and their composition. Numerical analyses made it possible to evaluate the mechanical functionality of the different fat tissues considering the characteristic loading conditions, as compressive and shear actions. The results pointed out the actual mechanical relevance of IFP and KSF, while ASF exhibited different mechanical properties. Furthermore, the contribution of connective septa and adipose lobules to compressive and shear mechanical behavior was elucidated. This preliminary investigation represents the basis for biomechanical interpretation and the definition of more refined model to be developed on the acquisition of additional histological and morphometric data.

Saccharomyces boulardii CNCM I-745 supplementation reduces gastrointestinal dysfunction in an animal model of IBS.

BACKGROUND: We evaluated the effect of Saccharomyces boulardii CNCM I-745 on intestinal neuromuscular anomalies in an IBS-type mouse model of gastrointestinal motor dysfunctions elicited by Herpes Simplex Virus type 1 (HSV-1) exposure. METHODS: Mice were inoculated intranasally with HSV-1 (102 PFU) or vehicle at time 0 and 4 weeks later by the intragastric (IG) route (108 PFU). Six weeks after IG inoculum, mice were randomly allocated to receive oral gavage with either S. boulardii (107 CFU/day) or vehicle. After 4 weeks the following were determined: a) intestinal motility using fluorescein-isothiocyanate dextran distribution in the gut, fecal pellet expulsion, stool water content, and distal colonic transit of glass beads; b) integrity of the enteric nervous system (ENS) by immunohistochemistry on ileal whole-mount preparations and western blot of protein lysates from ileal longitudinal muscle and myenteric plexus; c) isometric muscle tension with electric field and pharmacological (carbachol) stimulation of ileal segments; and d) intestinal inflammation by levels of tumor necrosis factor α, interleukin(IL)-1ß, IL-10 and IL-4. RESULTS: S. boulardii CNCM I-745 improved HSV-1 induced intestinal dysmotility and alteration of intestinal transit observed ten weeks after IG inoculum of the virus. Also, the probiotic yeast ameliorated the structural alterations of the ENS induced by HSV-1 (i.e., reduced peripherin immunoreactivity and expression, increased glial S100ß protein immunoreactivity and neuronal nitric oxide synthase level, reduced substance P-positive fibers). Moreover, S. boulardii CNCM I-745 diminished the production of HSV-1 associated pro-inflammatory cytokines in the myenteric plexus and increased levels of anti-inflammatory interleukins. CONCLUSIONS: S. boulardii CNCM I-745 ameliorated gastrointestinal neuromuscular anomalies in a mouse model of gut dysfunctions typically observed with irritable bowel syndrome.

Double origin of the superior cerebellar artery associated with homolateral haemorrhagic infarction of cerebellum.

The superior cerebellar artery (SCA) shows the least variable course and the lowest incidence of anatomical variations among cerebellar arteries. In the present study, an 84-year-old woman was affected by a cerebellar infarction which underwent haemorrhagic evolution in the following days. Neuroimaging investigations also showed a probable double origin of the left SCA. Neuropathological examination confirmed the presence of a large haemorrhagic infarction at the level of the superior portion of the left cerebellar hemisphere and vermis. The left SCA arose from two small arteries arising from the left aspect of the basilar artery and joining together after a course of 9 mm. Previous studies have reported the association of cerebrovascular pathologies, such as intracranial aneurysms, with fenestrations and double origins of the posterior inferior cerebellar artery. In the present case, the occurrence of an haemorrhagic infarction in the vascular field of an SCA with double origin is intriguing in suggesting a possible pathophysiological association.

Effects of postnatal hyperoxia exposure on the rat dentate gyrus and subventricular zone.

Premature newborns may be exposed to hyperoxia in the first postnatal period, but clinical and experimental works have raised the question of oxygen toxicity for the developing brain. However, specific analysis of hyperoxia exposure on neurogenesis is still lacking. Thus, the aim of the present study was to evaluate possible changes in the morphometric parameters of the main neurogenic sites in newborn rats exposed to 60 or 95 % oxygen for the first 14 postnatal days. The optical disector, a morphometric method based upon unbiased sampling principles of stereology, was applied to analyse cell densities, total volumes, and total cell numbers of the dentate gyrus (DG) and subventricular zone (SVZ). Apoptosis and proliferation were also studied by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling method and anti-ki67 immunohistochemistry, respectively. Severe hyperoxia increased the percentage of apoptotic cells in the DG. Moderate and severe hyperoxia induced a proliferative response both in the DG and SVZ, but the two neurogenic sites showed different changes in their morphometric parameters. The DG of both the hyperoxic groups showed lower volume and total cell number than that of the normoxic one. Conversely, the SVZ of newborn rats exposed to 95 % hyperoxia showed statistically significant higher volume and total cell number than SVZ of rats raised in normoxia. Our findings indicate that hyperoxia exposure in the first postnatal period affects both the neurogenic areas, although in different ways, i.e. reduction of DG and expansion of SVZ.

Cyclosporine and hyperoxia-induced lung damage in neonatal rats.

Cyclosporine effects on hyperoxia-induced histopathological and functional changes in the rat adult lung are controversial and the newborn lung has not been studied. Thus, we evaluated the effects of cyclosporine in young rats after 60% hyperoxia exposure postnatally. Experimental categories included: (1) room air for the first 5 postnatal weeks with daily subcutaneous injections of saline from postnatal day (PN)15 to PN35; (2) room air with daily injections of cyclosporine from PN15 to PN35; (3) 60% oxygen from PN0 to PN14 and then daily saline injections during the following three weeks; (4) 60% oxygen from PN0 to PN14 followed by cyclosporine treatment from PN15 to PN35. Hyperoxia significantly reduced the number of secondary crests and microvessel density, and it increased the mean alveolar size and septa thickness. Cyclosporine treatment did not significantly modify the hyperoxia-induced changes. Conversely, in normoxia, cyclosporine reduced microvessel density and the number of secondary crests. In conclusion, cyclosporine did not modify alveolar and microvascular parameters in hyperoxia exposure, although it caused some changes in normoxia.

Quality management of Body Donation Program at the University of Padova.

Quality management improvement has become a recent focus of attention in medical education. The program for the donation of bodies and body parts (Body Donation Program) at the University of Padova has recently been subjected to a global quality management standard, the ISO 9001:2008 certification. The aim of the present work is to show how the above standard is useful in enhancing the efficiency of body donation procedures and the quality and output of medical education. The program is managed by means of the following interlinked procedures: the collection of body donations, death certificates, data, and body parts from living donors; the transportation and identification of cadavers; the management of bodies, body parts, equipment, instruments, purchasing of necessary materials, and setting up anatomical training sessions; the management of preventive and corrective actions; the management of documents and registration; the management of internal and external quality audits; and the review of outcomes and improvement planning. Monitoring indicators are identified in the numbers of donors and of donated body parts per year, education sessions, and satisfaction of learners and donors, as evaluated by questionnaires. The process management approach, the integrated involvement of medical, technical, and administrative staff in defining procedures, and the application of monitoring indicators allow quality improvement in all aspects of the Body Donation Program.

Detection of apoptosis in human brainstem by TUNEL assay.

The aim of the present study was to evaluate the distribution of apoptotic neurons in the nuclei of the brainstem. Apoptosis was studied by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay, in human brainstems taken at autopsy from 15 adults (age range: 25-58 years). The nuclei examined included the hypoglossal nucleus (XII), dorsal motor nucleus of the vagus (DMNV), solitary tract nucleus (STN), vestibular nucleus (Ve), cuneate nucleus (Cu), nucleus of the spinal trigeminal tract (NSTT), principal inferior olivary nucleus (PION), medial inferior olivary nucleus (MION) and dorsal inferior olivary nucleus (DION). For each nucleus, the apoptotic index was expressed as the percentage of TUNEL-positive neurons out of the total number of neurons counted. Statistical analysis was performed with one-way ANOVA and Student-Newman-Keuls test for multiple comparisons. One-way ANOVA revealed that the differences in apoptotic index among the nuclei were statistically significant (P<0.001). Cu had the highest index, corresponding to 29.5 +/- 14.2%. The Student-Newman-Keuls test revealed significant differences when this value was compared with those of XII (14.1 +/- 12.0%; P<0.05), PION (12.8 +/- 13.8%; P<0.01), DION (12.0 +/-12.4%; P<0.01), MION (12.0 +/- 15.7%; P<0.01), STN (8.5 +/- 11.9%; P<0.001) and DMNV (5.4 +/- 6.8%; P<0.001). The differences in apoptotic index between Cu and NSTT (27.1 +/- 18.1%) and between Cu and Ve (23.7 +/- 12.6%) were not statistically significant (P>0.05). The apoptotic index in the NSTT was higher than in DMNV (P<0.001), STN (P<0.01) and nuclei of the inferior olivary complex (P<0.05). Ve showed a higher apoptotic index with respect to DMNV (P<0.01) and STN (P<0.05). Apoptotic processes found in the brainstems were ascribed to hypoxic-ischemic injury, and heterogeneity in apoptotic index among the nuclei supports the hypothesis of the differing vulnerability of the nuclei of the brainstem.