Co-administration of conjugated linoleic acid and rosiglitazone increases atherogenic co-efficient and alters isoprenaline-induced vasodilatation in rats fed high fat diet.

The cis(c)-9, trans(t)-11 (c9,t11) and t10,c12 isomers of conjugated linoleic acid (CLA) have been reported as agonists of peroxisome proliferator-activated receptor (PPAR) and beneficial in lipidemia and glycemia. However, it is unclear whether CLA isomers enhance or antagonize effects of conventional drugs targeting PPAR. Male Sprague-Dawley rats were fed high fat diet (HFD) for 8 weeks and treated without or with CLA, rosiglitazone or both for 4 weeks. Oral glucose tolerance and surrogate markers of insulin resistance were not significantly different for all treatments compared to untreated normal diet (ND) or HFD group, except lipoprotein levels. The combination of CLA and rosiglitazone had suppressed levels of low and high density lipoproteins (46 % and 25 %, respectively), compared to HFD-alone. Conversely, the atherogenic co-efficient of the animals received HFD or HFD+rosiglitazone+CLA was 2-folds higher than ND, HFD+rosiglitazone or HFD+CLA. Isolated aortic rings from the combined CLA and rosiglitazone treated animals were less sensitive to isoprenaline-induced relaxation among endothelium-denuded aortas with a decreased efficacy and potency (R(max)=53+/-4.7 %; pEC50=6+/-0.2) compared to endothelium-intact aortas (R(max)=100+/-9.9 %; pEC50=7+/-0.2). Our findings illustrate that the combination of CLA and rosiglitazone precede the atherogenic state with impaired endothelium-independent vasodilatation before the onset of HFD-induced insulin resistance.

Evaluation of the new red cell parameters on Beckman Coulter DxH800 in distinguishing iron deficiency anaemia from thalassaemia trait.

INTRODUCTION: The new red blood cell (RBC) parameters such as reticulocyte haemoglobin content and percentage of hypochromic red cells or equivalent, although useful in the laboratory assessment of iron deficiency anaemia (IDA), are confounded by thalassaemia trait (TT). We aim to evaluate the new red cell parameters on the Beckman Coulter DxH800 in distinguishing between IDA and TT. METHODS: A total of 246 normal subjects, 102 patients with IDA and 115 subjects with TT were accrued for the study. The parameters studied were red blood cell size factor (RSF), low haemoglobin density (LHD%), microcytic anaemia factor (MAF), standard deviation of conductivity of the nonreticulocyte population (SD-C-NRET) and unghosted cell (UGC). Comparison between groups was performed by Student's t-test, and the diagnostic performance was determined by receiver operating characteristic (ROC) curve analysis. RESULTS: Both the LHD% and RSF were significantly higher in IDA than TT, whereas MAF and SD-C-NRET were significantly lower. The SD-C-NRET showed the best diagnostic performance as a single parameter. A formula, [(RBC + Hb) × (HCT + SD-C-NRET)]/RDW-SD, was devised to distinguish between IDA and TT. With a cut-off value of 23, the area under the curve (AUC) was 0.995 (95% CI of 0.99-1.00), the sensitivity was 97%, and the specificity was 99.1%. CONCLUSIONS: The new RBC parameters on Beckman Coulter DxH800 provide useful information in distinguishing between IDA and TT, which is important for clinical decision-making and for streamlining laboratory testing. A new formula is devised that performs better than other discriminant functions in the literature.

Apocynum venetum leaf extract, an antihypertensive herb, inhibits rat aortic contraction induced by angiotensin II: a nitric oxide and superoxide connection.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves extract of Apocynum venetum (AVLE), also known as "luobuma", have long been used in traditional Chinese medicine to treat hypertension and depression in parts of China and it has been shown to possess anti-oxidant and anti-lipid peroxidation effects. AVLE (10 µg/ml) has been reported to have a long-lasting endothelium-dependent relaxant effect and this effect has been proposed to be due to its nitric oxide(NO)-releasing and superoxide anion(SOA)-scavenging properties. AIM OF THE STUDY: The present study seeks to evaluate the differential actions of AVLE extract between Ang II- and PE-induced vasoconstriction and the involvement of superoxide anions. MATERIALS AND METHODS: Single dose of Ang II (100 nM and 1 nM)- or PE (0.1 µM)-induced contraction were assessed in both endothelium-intact and -denuded aortic rings after pre-incubation of AVLE (10 µg/ml) for 15 min. The experiment was repeated in either the presence of NO synthase inhibitor, L-NAME (300 µM) or selective AT(1) receptor inhibitor, losartan (0.1 nM), or superoxide scavenger, tiron (1 mM) or a combination of L-NAME and AVLE. Superoxide production was measured by using enhanced-chemiluminescence assay. RESULTS: We have demonstrated that AVLE (10 µg/ml) effectively suppressed the Ang II-induced contraction (100 nM and 1 nM) of both endothelium-intact and -denuded rat aortic rings. In endothelium-intact rings, L-NAME, reversed AVLE-induced inhibition of Ang II-contraction. PE-induced contraction was significantly inhibited by AVLE in endothelium-intact rings, but not in endothelium-denuded rings. The inhibition by AVLE of PE-induced contraction was totally abolished in the presence of L-NAME. Ang II-induced SOA production concentration dependently with the optimal effect seen at 100 nM of Ang II, and AVLE (0.3, 1, 10 µg/ml) reduced this effect. SOA production in Ang II-stimulated rings was significantly higher than unstimulated control rings, while PE did not stimulate SOA production at all. SOA formation in the presence of Ang II was also inhibited in the presence of SOD (superoxide scavenger), DPI (NADPH inhibitor) and losartan (specific AT(1) receptor antagonist). CONCLUSION: These results collectively suggest that the ability of AVLE in inhibiting Ang II-induced contraction via its SOA scavenging properties and nitric oxide releasing effect may account for its usage as an antihypertensive treatment in traditional folk medicine.

Osteoclast-like cells in soft tissue leiomyosarcomas.

Giant cell-rich leiomyosarcoma of soft tissues is an unusual variant of malignant smooth muscle tumor characterized by the presence of numerous multinucleated giant cells (MNGCs). The nature of MNGCs and the cellular mechanisms underlying their accumulation in this tumor are poorly understood. Analysis of the expression of osteoclast, macrophage, and smooth muscle markers in two cases of giant cell-rich leiomyosarcoma revealed that the MNGCs in giant cell-rich leiomyosarcoma were negative for smooth muscle markers and that these cells expressed an osteoclast-like phenotype, being positive for CD45, CD68, tartrate-resistant acid phosphatase, and CD51 but negative for CD14 and HLA-DR. Scattered tumor-associated macrophages (TAMs) also expressed this phenotype. Leiomyosarcoma tumor cells strongly reacted for CD51 but were negative for CD14, CD45, and CD68. An analysis of 25 conventional (nongiant cell-containing) leiomyosarcomas found isolated CD68(+) MNGCs in three cases (12%), all of which were grade II/III leiomyosarcomas containing a prominent TAM infiltrate. Leiomyosarcoma-derived TAMs in the presence of receptor activator for nuclear factor kappa B ligand (RANKL) and macrophage colony-stimulating factor were capable of differentiating into osteoclast-like cells capable of resorbing bone. Reverse transcription polymerase chain reaction studies showed that RANKL, osteoprotegerin, and TNF-related apoptosis-inducing ligand were expressed by leiomyosarcoma cells. Our findings indicate that the giant cells found in leiomyosarcomas are osteoclast-like and that they are formed from TAMs by a RANKL-dependent mechanism.

Treatment of soft tissue contour defects by a combination of surgical subcision with a Beaver tympanoplasty blade and autologous fat grafting.

This report describes a technique for treating depressed scars and soft tissue contour deformities. Surgical subcision with a Beaver tympanoplasty blade is undertaken for depressed or adherent scars to release the fibrous attachments beneath the scar before autologous fat grafting. Satisfactory results were observed, with an improvement in surface contour for 16 patients over a 3-year period. The authors recommend the described technique as a safe, minimally invasive, and precise method for subcutaneous dissection of scar tissue before fat injections.

Endurance exercise promotes cardiorespiratory rehabilitation without neurorestoration in the chronic mouse model of parkinsonism with severe neurodegeneration.

Physical rehabilitation with endurance exercise for patients with Parkinson's disease has not been well established, although some clinical and laboratory reports suggest that exercise may produce a neuroprotective effect and restore dopaminergic and motor functions. In this study, we used a chronic mouse model of Parkinsonism, which was induced by injecting male C57BL/6 mice with 10 doses of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (25 mg/kg) and probenecid (250 mg/kg) over 5 weeks. This chronic parkinsonian model displays a severe and persistent loss of nigrostriatal neurons, resulting in robust dopamine depletion and locomotor impairment in mice. Following the induction of Parkinsonism, these mice were able to sustain an exercise training program on a motorized rodent treadmill at a speed of 18 m/min, 0 degrees of inclination, 40 min/day, 5 days/week for 4 weeks. At the end of exercise training, we examined and compared their cardiorespiratory capacity, behavior, and neurochemical changes with that of the probenecid-treated control and sedentary parkinsonian mice. The resting heart rate after 4 weeks of exercise in the chronic parkinsonian mice was significantly lower than the rate before exercise, whereas the resting heart rate at the beginning and 4 weeks afterward in the control or sedentary parkinsonian mice was unchanged. Exercised parkinsonian mice also recovered from elevated electrocardiogram R-wave amplitude that was detected in the parkinsonian mice without exercise for 4 weeks. The values of oxygen consumption, carbon dioxide production, and body heat generation in the exercised parkinsonian mice before and during the Bruce maximal exercise challenge test were all significantly lower than that of their sedentary counterparts. Furthermore, the exercised parkinsonian mice demonstrated a greater mass in the left ventricle of the heart and an increased level of citrate synthase activity in the skeletal muscles. The amphetamine-induced, dopamine release-dependent locomotor activity was markedly inhibited in the sedentary parkinsonian mice and was also inhibited in the exercised parkinsonian mice. Finally, neuronal recovery from the loss of nigrostriatal tyrosine hydroxylase expression and dopamine levels in the severe parkinsonian mice after exercise was not evident. Taken all together, these data suggest that 4 weeks of treadmill exercise promoted physical endurance, resulting in cardiorespiratory and metabolic adaptations in the chronic parkinsonian mice with severe neurodegeneration without demonstrating a restorative potential for the nigrostriatal dopaminergic function.

Cellular and humoral mechanisms of osteoclast formation in Ewing's sarcoma.

Cellular mechanisms that account for tumour osteolysis associated with Ewing's sarcoma are uncertain. Osteoclasts are marrow-derived multinucleated cells (MNCs) that effect tumour osteolysis. Osteoclasts are known to form from macrophages by both receptor activator for nuclear factor-kappaB (RANK) ligand (RANKL)-dependent and -independent mechanisms. In this study, our aim has been to determine whether tumour-associated macrophages (TAMs) isolated from Ewing's sarcoma are capable of differentiating into osteoclasts and to characterise the cellular and humoral mechanisms whereby this occurs. Tumour-associated macrophages were isolated from two Ewing's sarcomas and cultured on both coverslips and dentine slices for up to 21 days with soluble RANKL and macrophage colony stimulating factor (M-CSF). Osteoclast formation from TAMs (CD14+) was evidenced by the formation of tartrate-resistant acid phosphatase (TRAP) and vitronectin receptor (VNR)-positive MNCs, which were capable of carrying out lacunar resorption. This osteoclast formation was inhibited by the addition of bisphosphonates. Both Ewing's sarcoma-derived fibroblasts and some bone stromal cells expressed RANKL and supported osteoclast formation by a contact-dependent mechanism. We also found that osteoclast differentiation occurred when Ewing's TAMs were cultured with tumour necrosis factor (TNF)-alpha in the presence of M-CSF and that TC71 Ewing's sarcoma cells stimulated osteoclast formation through the release of a soluble factor, the action of which was abolished by an antibody to TNF-alpha. These results indicate that TAMs in Ewing's sarcoma are capable of osteoclast differentiation by both RANKL-dependent and TNF-alpha-dependent mechanisms and that Ewing's sarcoma cells produce osteoclastogenic factor(s). Our findings suggest that anti-resorptive and anti-osteoclastogenic therapies may be useful in inhibiting the osteolysis of Ewing's sarcoma.

RANKL-dependent and RANKL-independent mechanisms of macrophage-osteoclast differentiation in breast cancer.

The cellular and humoral mechanisms accounting for tumour osteolysis in metastatic breast cancer are uncertain. Osteoclasts, the specialised multinucleated cells responsible for tumour osteolysis, are derived from monocyte/macrophage precursors. Breast cancer-derived tumour-associated macrophages (TAMs) are capable of osteoclast differentiation but the cellular and humoral mechanisms controlling this activity are uncertain. In this study, TAMs were isolated from primary breast cancers and cultured in the presence and absence of cytokines/growth factors influencing osteoclastogenesis. Extensive TAM-osteoclast differentiation occurred only in the presence of RANKL and M-CSF; this process was inhibited by OPG and RANK:Fc, decoy receptors for RANKL. Breast cancer-derived fibroblasts and human bone stromal cells expressed mRNA for RANKL, OPG and TRAIL, and co-culture of these fibroblasts with human monocytes stimulated osteoclast formation by a RANKL-dependent mechanism. Osteoclast formation and lacunar resorption also occurred by a RANKL-independent mechanism when the conditioned medium from breast cancer cells, MDA-MB-231 and MCF-7, was added (with M-CSF) to monocyte cultures. Our findings indicate that TAMs in breast cancer are capable of osteoclast differentiation and that breast cancer-derived fibroblasts and breast cancer cells contribute to this process by producing soluble factors that influence osteoclast formation by RANKL-dependent and RANKL-independent mechanisms respectively.

An insight into burns in a developing country: a Sri Lankan experience.

Burn injuries represent a diverse and varied challenge to medical and paramedical staff. The management of burns and their sequelae in a well-equipped, modern burns unit remains demanding despite advances in surgical techniques and development of tissue-engineered biomaterials; in a developing country, these difficulties are amplified many times. Sri Lanka has a high incidence of burn-related injuries annually due to a combination of adverse social, economic and cultural factors. The management of burn injuries remains a formidable public health problem. The epidemiology of burns, challenges faced in their management and effective strategies specific to Sri Lanka, such as the Safe Bottle Lamp campaign, are highlighted in this paper.

Malignant melanoma and bone resorption.

The cellular and humoral mechanisms accounting for osteolysis in skeletal metastases of malignant melanoma are uncertain. Osteoclasts, the specialised multinucleated cells that carry out bone resorption, are derived from monocyte/macrophage precursors. We isolated tumour-associated macrophages (TAMs) from metastatic (lymph node/skin) melanomas and cultured them in the presence and absence of osteoclastogenic cytokines and growth factors. The effect of tumour-derived fibroblasts and melanoma cells on osteoclast formation and resorption was also analysed. Melanoma TAMs (CD14+/CD51-) differentiated into osteoclasts (CD14-/CD51+) in the presence of receptor activator for nuclear factor kappaB ligand (RANKL) and macrophage-colony stimulating factor. Tumour-associated macrophage-osteoclast differentiation also occurred via a RANKL-independent pathway when TAMs were cultured with tumour necrosis factor-alpha and interleukin (IL)-1alpha. RT-PCR showed that fibroblasts isolated from metastatic melanomas expressed RANKL messenger RNA and the conditioned medium of cultured melanoma fibroblasts was found to be capable of inducing osteoclast formation in the absence of RANKL; this effect was inhibited by the addition of osteoprotegerin (OPG). We also found that cultured human SK-Mel-29 melanoma cells produce a soluble factor that induces osteoclast differentiation; this effect was not inhibited by OPG. Our findings indicate that TAMs in metastatic melanomas can differentiate into osteoclasts and that melanoma fibroblasts and melanoma tumour cells can induce osteoclast formation by RANKL-dependent and RANKL-independent mechanisms, respectively.

Early signs of neuronal apoptosis in the substantia nigra pars compacta of the progressive neurodegenerative mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model of Parkinson's disease.

Parkinson's disease is associated with a progressive loss of substantia nigra pars compacta dopaminergic neurons. The cellular and molecular mechanisms underlying Parkinson's disease neurodegeneration have not been fully determined. Clinical investigations and subacute in vivo studies using the neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine have generated some observations suggesting that apoptosis is involved in neurodegeneration; however, this view remains equivocal. In this study, the substantia nigra pars compacta neurodegenerative process was examined in the chronic mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model of Parkinson's disease treated with 10 doses of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (25 mg/kg) and probenecid (250 mg/kg) over five weeks. One day after chronic treatment, numerous terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells were detected specifically in the substantia nigra pars compacta displaying shrunken volume, chromatin condensation, and DNA fragmentation. The number of apoptotic cells declined over time. No terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells were found in untreated or probenecid-treated control animals. Cytomorphometric analysis of substantia nigra pars compacta nuclear loci revealed eccentric nucleoli dislocation and vesicular degranulation in all of the apoptotic neurons for the mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model for Parkinson's disease. The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells phenotypically showed neuronal origin (NeuN-positive) with a loss of tyrosine hydroxylase immunoreactivity. While the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells were not co-localized with astroglial (GFAP-positive) cells, some apoptotic cells were clearly associated with the activated microglial (macrophage antigen complex-1 and isolectin B(4)-positive) cells suggesting an active process of dead cell removal. In the one-day and seven-day post-treated mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model for Parkinson's disease, marked depression of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and striatum was observed, which was correlated with significant reductions of striatal dopamine content and uptake. These results suggest that initial neuronal apoptosis and morphological changes are involved, at least in part, in the chronic neurodegeneration of mouse 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid model for Parkinson's disease.

Bone stromal cells in pagetic bone and Paget's sarcoma express RANKL and support human osteoclast formation.

Paget's disease is a focal disorder of bone remodelling, in which there is an increase in osteoclast formation. A rare complication of Paget's disease is the development of a sarcoma, most commonly an osteosarcoma. Osteoclast formation occurs in the presence of macrophage-colony stimulating factor and receptor activator for nuclear factor-kappaB ligand (RANKL), and it has been shown that bone stromal cells in Paget's disease can influence osteoclast formation by modulating the expression of RANKL and its decoy receptor, osteoprotegerin (OPG). In this study we show that pagetic bone stromal cells express RANKL and that these cells promote osteoclast formation by a RANKL-dependent mechanism. Osteoclast formation in co-cultures of monocytes and either pagetic bone stromal cells or Paget's sarcoma stromal cells was not only induced by a contact-dependent mechanism but also occurred via the release of a soluble factor. In contrast to bone stromal cells isolated from normal controls, stromal cells isolated from morphologically normal bone in one patient with Paget's disease also stimulated osteoclast formation in this way; this osteoclastogenesis was inhibited by OPG. Our results indicate that Paget's bone stromal cells support osteoclast formation by a RANKL-dependent process which involves not only cell-cell contact but also secretion of soluble RANKL.

Surgical treatment of in-growing toenails performed by senior house officers: are they good enough?

BACKGROUND: Although the surgical treatment of in-growing toenails is a common procedure, the success of ingrown toenail surgery is extremely variable and recurrences frequently impair the quality of lift of those who have this condition. In most hospitals this procedure is commonly performed by junior surgical trainees who may have little experience. AIM: We proposed to find out if the success of this procedure is operator-dependent by comparing the results of standard of toenail surgery performed by basic surgical trainees (BSTs) in our hospitals with already published data. METHODS: A retrospective analysis of nail bed ablation surgery performed by BSTs under local anaesthesia over a 15-month period in a district general hospital was conducted. RESULTS: 106 phenol ablations and 46 germinal matrix excisions were prformed. Symptomatic recurrence rates 12 months following the procedure were 5.7% for phenol ablation and 4.3% for germinal matrix excisions. CONCLUSION: Our results are comparable to published data, and we conclude that toenail ablation surgery can be just as successfully performed by junior surgeons after relatively little training in the procedure.

Audit of plastic surgeons' understanding of pathology reports of skin neoplasia.

This audit was set up to investigate Plastic Surgeons' perception of minimum standards of reporting of skin neoplasia at the time of publication of the Royal College of Pathologists' guidelines. Eight histopathology case reports of skin neoplasia were formulated; four had the minimum information required and four had vital information omitted. Surgeons were asked to evaluate and suggest management. Marks were allocated for awareness of substandard reporting and for patient management. Forty-three out of 60 questionnaires were returned. Consultants had a significantly higher total score compared to the other grades (p<0.05) A high knowledge score was correlated with a high management score. However, awareness of minimum data was relatively poor across all grades. The majority of participants themselves were unaware of the new guidelines (38/43). Awareness of the guidelines and increased understanding of basic pathology should correct these weaknesses.

Hypercytolipidemia-induced nuclear lipoapoptosis: cytochemical analysis and integrated review of hypogonadal, diabetes-obesity syndrome-induced female reproductive axis disruption.

Expression of the diabetes (db/db) mutation (i.e., leptin receptor defect) in C57BL/KsJ mice results in the functional suppression of the female pituitary-gonadal axis accompanied by premature utero-ovarian lipocytoatrophy. The current studies define the cytostructural, metabolic and endocrine disturbances associated with hypercytolipidemia and coincident nuclear lipoapoptosis following expression of the db/db-mutation. Adult, female C57BL/KsJ control (+/+ and +/? genotypes) and db/db mutant littermates were monitored for systemic alterations in blood glucose, insulin, luteinizing hormone (LH) and 17-B-estradiol (E2) concentrations associated with db/db-enhanced cytolipid depositions and TUNEL-labeled 3'-DNA fragmentation indexed nuclear lipoapoptosis. Obesity, hyperglycemia and hyperinsulinemia, in addition to depressed LH and E2 concentrations, characterized all db/db-mutants relative to control indices. Structural and cytochemical analysis of basophilic gonadotroph cells, ovarian follicular granulosa cells and uterine endometrial epithelial layers indicated that db/db mutants demonstrated prominent hypercytolipidemia relative to control cytoarchitecture profiles. Vasolipidemia and interstitial cytoadiposity were prominent in all db/db tissue compartments. In each affected cell type within the db/db pituitary-reproductive tract axis, hypercytolipidemia was localized with pronounced nuclear lipo-infiltration and 3'-DNA TUNEL-labeled fragmentation. These data indicate that coincident cytostructural, endocrine and metabolic disturbances associated with hypogonadal pituitary-reproductive tract hypercytolipidemia are functional manifestations of the expressed diabetes-obesity syndrome in db/db-mutants. The progressive vaso-, interstitial-, and cyto-lipidemic alterations in cytoarchitecture correlated with the coincident nuclear lipoapoptotic dissolution and pronounced organo-involution, alterations which contributed to the functional disruption of the pituitary-hypogonadal axis in C57BL/KsJ-db/db mice.

Hypophyseal lipoapoptosis: diabetes (db/db) mutation-associated cytolipidemia promotes pituitary cellular disruption and dysfunction.

Expression of the diabetes (db/db) mutation in C57BL/KsJ mice suppresses the female pituitary-gonadal axis via progressive cytolipidemic disruption of hypophyseal gonadotropin release, culminating in premature involution of the reproductive tract and manifest infertility. The current studies define the systemic, endocrine, cytochemical and structural apoptotic changes that result from pituitary hypercytolipidemia induced by db/db mutation expression in this Type II diabetes-obesity syndrome (DOS) model. Adult female C57BL/KsJ control (+/? genotype) and db/db littermates were monitored for systemic and cellular alterations in LH-, FSH- and gonadal steroid-secretion, and coincident pituitary apoptosis, as indexed by TUNEL labeled 3' nuclear DNA-fragmentation, associated with cytolipid depositions. Obesity, hyperglycemia and hyperinsulinemia characterized all db/db-mutants relative to +/? groups. Serum progesterone (P) and estradiol (E2) concentrations were suppressed in db/db mutants coincident with decreased plasma LH and FSH concentrations relative to +/? values. Cytochemical analysis of anterior (AP) pituitary cell subtypes indicated that db/db mutants demonstrated prominent hypercytolipidemia relative to +/? pituitary cytoarchitecture. Cytolipidemic vacuoles were localized within protein vesiculated db/db hypophyseal basophilic and acidophilic cell populations. Hypophyseal cytoadiposity in db/db AP cells was co-localized with prominent cellular apoptotic TUNEL labeling of nuclear 3'-DNA fragments in cells demonstrating vesicular depopulation and cytolytic vacuolization. These data represent the first demonstration of co-localized hypercytolipidemic and cytoapoptotic disruptive events occurring concurrently in a hypopituitary-hypogonadal syndrome model following expression of the Type II (NIDDM) diabetes-obesity syndrome in db/db-mutants. The coincident and progressive vascular-, interstitial- and cyto-lipidemic alterations in hypophyseal cytoarchitecture correlated with the concurrent apoptotic disruption of pituitary endocrine cytoarchitecture and supressed gonadal steroid synthesis, influences which collectively contribute to the premature involution of the pituitary-gonadal axis in C57BL/KsJ- db/db mice.

Lysosomal malfunction accompanies alpha-synuclein aggregation in a progressive mouse model of Parkinson's disease.

We have detected granular and filamentous inclusions that are alpha-synuclein- and ubiquitin-immunoreactive in the cytoplasm of dopaminergic and cortical neurons of C57/black mice treated chronically with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and probenecid. The immunoreactive aggregates only become evident several weeks after large-scale dopaminergic cell death and a downregulation of alpha-synuclein gene expression. Numerous lipofuscin granules accumulate alpha-synuclein in the nigral and limbic cortical neurons of treated mice. These data provide evidence that insoluble proteins, such as alpha-synuclein, build up as granular and filamentous inclusions in dopaminergic neurons that survive the initial toxic MPTP insult. They further suggest that defective protein degradation rather than altered gene expression underlies deposition of alpha-synuclein and that abundant lysosomal compartments are present to seal off the potentially toxic material.