Vascular units as advanced living materials for bottom-up engineering of perfusable 3D microvascular networks.

The timely establishment of functional neo-vasculature is pivotal for successful tissue development and regeneration, remaining a central challenge in tissue engineering. In this study, we present a novel (micro)vascularization strategy that explores the use of specialized "vascular units" (VUs) as building blocks to initiate blood vessel formation and create perfusable, stroma-embedded 3D microvascular networks from the bottom-up. We demonstrate that VUs composed of endothelial progenitor cells and organ-specific fibroblasts exhibit high angiogenic potential when embedded in fibrin hydrogels. This leads to the formation of VUs-derived capillaries, which fuse with adjacent capillaries to form stable microvascular beds within a supportive, extracellular matrix-rich fibroblastic microenvironment. Using a custom-designed biomimetic fibrin-based vessel-on-chip (VoC), we show that VUs-derived capillaries can inosculate with endothelialized microfluidic channels in the VoC and become perfused. Moreover, VUs can establish capillary bridges between channels, extending the microvascular network throughout the entire device. When VUs and intestinal organoids (IOs) are combined within the VoC, the VUs-derived capillaries and the intestinal fibroblasts progressively reach and envelop the IOs. This promotes the formation of a supportive vascularized stroma around multiple IOs in a single device. These findings underscore the remarkable potential of VUs as building blocks for engineering microvascular networks, with versatile applications spanning from regenerative medicine to advanced in vitro models.

Factors predicting non-ventilated hospital-acquired pneumonia: systematic review and meta-analysis.

BACKGROUND: Hospital-acquired pneumonia (HAP) results in approximately 15-20% of all infections in hospitals, with more than two-thirds being in patients not using mechanical ventilation. The incidence of non-ventilated hospital-acquired pneumonia (NVHAP) is increasing, and it is associated with a longer length of stay, the need for intensive care unit hospitalization and mechanical ventilation use, and higher mortality. AIM: To identify, quantify, and summarize predictive factors for NVHAP in adult patients admitted to non-intensive care units as determined by previous observational studies. METHODS: PubMed, Embase, Scopus, and LILACS were systematically searched. Case-control and cohort studies were included, and a meta-analysis was performed for all factors studied more than once. National Institute of Health assessment tools were applied to assess the quality of the studies. FINDINGS: Thirty-eight articles showing 204 predictive factors were included. A meta-analysis was performed for 58 factors, 32 of which were significantly associated with NVHAP. When the sensitivity analysis was performed without poor-quality studies, 24 factors remained associated with NVHAP. CONCLUSION: Although there is a lack of good-quality studies to establish predictive factors for NVHAP, the results of this study showed 24 factors associated with the development of this infectious complication. Knowledge of the significant predictive factors for NVHAP will enable the identification of patients most likely to develop it.

Propagation phase-contrast micro-computed tomography allows laboratory-based three-dimensional imaging of articular cartilage down to the cellular level.

OBJECTIVE: High-resolution non-invasive three-dimensional (3D) imaging of chondrocytes in articular cartilage remains elusive. The aim of this study was to explore whether laboratory micro-computed tomography (micro-CT) permits imaging cells within articular cartilage. DESIGN: Bovine osteochondral plugs were prepared four ways: in phosphate-buffered saline (PBS) or 70% ethanol (EtOH), both with or without phosphotungstic acid (PTA) staining. Specimens were imaged with micro-CT following two protocols: 1) absorption contrast (AC) imaging 2) propagation phase-contrast (PPC) imaging. All samples were scanned in liquid. The contrast to noise ratio (C/N) of cellular features quantified scan quality and were statistically analysed. Cellular features resolved by micro-CT were validated by standard histology. RESULTS: The highest quality images were obtained using propagation phase-contrast imaging and PTA-staining in 70% EtOH. Cellular features were also visualised when stained in PBS and unstained in EtOH. Under all conditions PPC resulted in greater contrast than AC (p < 0.0001 to p = 0.038). Simultaneous imaging of cartilage and subchondral bone did not impede image quality. Corresponding features were located in both histology and micro-CT and followed the same distribution with similar density and roundness values. CONCLUSIONS: Three-dimensional visualisation and quantification of the chondrocyte population within articular cartilage can be achieved across a field of view of several millimetres using laboratory-based micro-CT. The ability to map chondrocytes in 3D opens possibilities for research in fields from skeletal development through to medical device design and treatment of cartilage degeneration.

Synthesis, characterization and photocatalytic properties of nanostructured CoFe2O4 recycled from spent Li-ion batteries.

In this study, cobalt (Co) was recycled from spent lithium ion batteries (LIBs) and used to synthesize cobalt ferrite (CoFe2O4-LIBs), which was applied as a catalyst for heterogeneous photo Fenton reactions that discolored methylene blue (MB) dye. The co-precipitation method was used to synthesize CoFe2O4-LIBs and CoFe2O4-R nanoparticles with spinel structures using as raw materials of the LIB cathodes and commercial reagents. X-ray diffraction (XRD) identified the formation of spinel-type CoFe2O4, which formed clusters that could be seen under scanning electron microscopy (SEM) analysis and nanometric particles seen under transmission electron microscopy (TEM). Inductively Coupled Plasma Optical Emission Spectrometer (ICP OES) analysis was used to determine the concentrations of metals present in the ferrite, which reached 6.5% (w/w) of Co. The optimal conditions for discoloring the dye were evaluated using a factorial design. Using CoFe2O4 as a catalyst, the best conditions for catalytic reaction were pH 3, 30.0 mg of catalyst, and 8.0 mL of H2O2 73% (v/v). Discoloration efficiencies of 87.3% and 87.7% were obtained from CoFe2O4-R and CoFe2O4-LIBs, respectively. Therefore, CoFe2O4-LIBs proved to be an efficient catalyst for discoloring MB dye using heterogeneous photo-Fenton reactions. This work is of scientific, social, economic, and environmental interest. It investigates the process of synthesizing,characterizing CoFe2O4LIBs and the efficiency of degrading MB dye, subjects that have economic and environmental, and therefore, social interest. The work has scientific interest particularly because of the correlation between the structure of the recycled material and its catalytic properties.

Monomeric, porous type II collagen scaffolds promote chondrogenic differentiation of human bone marrow mesenchymal stem cells in vitro.

Osteoarthritis (OA) is a common cause of pain and disability and is often associated with the degeneration of articular cartilage. Lesions to the articular surface, which are thought to progress to OA, have the potential to be repaired using tissue engineering strategies; however, it remains challenging to instruct cell differentiation within a scaffold to produce tissue with appropriate structural, chemical and mechanical properties. We aimed to address this by driving progenitor cells to adopt a chondrogenic phenotype through the tailoring of scaffold composition and physical properties. Monomeric type-I and type-II collagen scaffolds, which avoid potential immunogenicity associated with fibrillar collagens, were fabricated with and without chondroitin sulfate (CS) and their ability to stimulate the chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells was assessed. Immunohistochemical analyses showed that cells produced abundant collagen type-II on type-II scaffolds and collagen type-I on type-I scaffolds. Gene expression analyses indicated that the addition of CS - which was released from scaffolds quickly - significantly upregulated expression of type II collagen, compared to type-I and pure type-II scaffolds. We conclude that collagen type-II and CS can be used to promote a more chondrogenic phenotype in the absence of growth factors, potentially providing an eventual therapy to prevent OA.

Expression patterns of α2,3-sialyltransferase I and α2,6-sialyltransferase I in human cutaneous epithelial lesions.

Skin tumors have become one of the most common cancers in the world and their carcinogenesis is frequently associated with altered glycosylation patterns. The aberrant sialylation, a type of glycosylation, can mediate pathophysiological key events during various stages of tumor progression, including invasion and metastasis. Sialyltransferases play a key role in a variety of biological processes, including cell-cell communication, cell-matrix interaction, adhesion, and protein targeting. In this study, it was evaluated the expression of ST3Gal I and ST6Gal I in cutaneous epithelial lesions that include actinic keratosis (n=15), keratoacanthoma (n=9), squamous cell carcinoma (n=22) and basal cell carcinoma (n=28) in order to evaluate if sialyltransferases expression is different in premalignant and in malignant tumors. The expression of ST3Gal I was observed in actinic keratosis (53%), keratoacanthoma (78%), squamous cell carcinoma (73%) and basal cell carcinoma (32%) with statistic differences between basal cell carcinoma and keratoacanthoma (P=0.0239) and basal cell carcinoma and squamous cell carcinoma (P=0.0096); for ST6Gal I, cytoplasmic expression was noted in actinic keratosis (40%), heterogeneous and cytoplasmic expression was noted in keratoacanthoma (67%), squamous cell carcinoma (41%) and basal cell carcinoma (7%) with statistic differences between basal cell carcinoma and squamous cell carcinoma (P=0.0061) and basal cell carcinoma and keratoacanthoma (P=0.0008). In summary, our results showed that the high expression of ST3Gal I and ST6Gal I, in skin tumors, is associated with tumors with greater potential for invasion and metastasis, as in the case of squamous cell carcinoma, and this may be related to their behavior.

Sialic acid differential expression in non-melanoma skin cancer biopsies.

Altered sialylation has been observed during oncogenic transformation and has been implicated in tumor progression and metastases. This pattern may aid the biological behavior of many tumors. Skin cancer is the most common cancer worldwide and their diagnosis becomes difficult, in some cases, due to variety of factors that affect the accuracy of the nowadays exams, such as huge spectrum of tumors and their variants. So, this study investigates the changes in expression and distribution of α2,3 and α2,6-linked sialic acid in non-melanomas skin cancer to identify the sialylation pattern which may be useful in the differential diagnosis of this tumor. Lectin histochemistry was used to examine the expression and distribution of sialic acid in different types of non-melanoma skin cancers. We applied Maackia amurensis lectin, which interacts with α2,3-linked sialic acid and Sambucus nigra lectin specific for α2,6-linked sialic acid. The histochemical analysis showed that α2,3 and α2,6-linked sialic acid vary their expression according with the tumor type analyzed. The distribution of α2,3-linked sialic was differentially expressed in between basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) (p < 0.0001), BCC and actinic keratosis (p = 0.0033) and BCC and keratoacanthoma (p < 0.0001). In the case of α2,6-linked sialic acid its expression was also different between BCC and SCC (p < 0.0001), BCC and actinic keratosis (p = 0.0002) and BCC and keratoacanthoma (p < 0.0362). Lectin histochemistry showed a different expression of both sialic acid linkages types between pre-malign and malign tumors and between malign tumors. Although preliminary, these findings are promising for the development of diagnostic techniques to help in the differential diagnosis of non-melanoma skin tumors using lectin histochemistry as an auxiliary tool.

Surveillance Programme for Healthcare Associated Infections in the State of São Paulo, Brazil. Implementation and the first three years' results.

Governmental programmes should be developed to collect and analyse data on healthcare associated infections (HAIs). This study describes the healthcare setting and both the implementation and preliminary results of the Programme for Surveillance of Healthcare Associated Infections in the State of São Paulo (PSHAISP), Brazil, from 2004 to 2006. Characterisation of the healthcare settings was carried out using a national database. The PSHAISP was implemented using components for acute care hospitals (ACH) or long term care facilities (LTCF). The components for surveillance in ACHs were surgical unit, intensive care unit and high risk nursery. The infections included in the surveillance were surgical site infection in clean surgery, pneumonia, urinary tract infection and device-associated bloodstream infections. Regarding the LTCF component, pneumonia, scabies and gastroenteritis in all inpatients were reported. In the first year of the programme there were 457 participating healthcare settings, representing 51.1% of the hospitals registered in the national database. Data obtained in this study are the initial results and have already been used for education in both surveillance and the prevention of HAI. The results of the PSHAISP show that it is feasible to collect data from a large number of hospitals. This will assist the State of São Paulo in assessing the impact of interventions and in resource allocation.

Low body mass index and declining sex steroids explain most age-related bone loss in Brazilian men.

SUMMARY: Osteoporosis in men is underestimated, but our data point to an increasing prevalence rate in those over 70 years old with body mass index (BMI) <25 kg/m(2), bioavailable testosterone <2.7 nmol/L, bioavailable estradiol <40 pmol/L, and high bone turnover, defined in this study as serum carboxyterminal cross-linked telopeptide of type I collagen (ICTP) >4.3 microg/L. INTRODUCTION: The association of sex steroids and osteoporosis was evaluated in 104 men, aged 50-93 years old. METHODS: Bone mineral density (BMD), bone turnover (ICTP), testosterone (T), and estradiol (E(2)) were measured; free and bioavailable hormones (free testosterone index [FTI], BioT, free estradiol index [FEI], and BioE(2)) were calculated from T, E(2), sex hormone-binding globulin (SHBG), and albumin. Nonparametric analysis and Poisson regression models were used. RESULTS: Significant increases in SHBG and ICTP and decreases in femoral neck BMD, FTI, FEI, BioT, and BioE(2) were observed with each additional decade of age. Femoral neck BMD was inversely correlated with ICTP, and both were significantly associated with SHBG, FTI, BioT, FEI, and BioE. There was a direct and graded association between age and osteoporosis prevalence rate (OP PR; p = 0.028). Compared to participants less than 70 years old, the crude OP PR of those 80 years and older was 3.2 (95%CI = 1.4-7.3). Adjusting sequentially for BMI and bioavailable sex hormones attenuated the association between age and osteoporosis prevalence by 55% and 77%, respectively. CONCLUSION: Our data support the view that low BMI and declining sex steroids explain most of the association between aging, increased bone turnover, and osteoporosis in men.

Detection of oncogenic human papillomavirus in sporadic retinoblastoma.

PURPOSE: To investigate the presence of human papillomavirus (HPV) DNA in tumour tissue from patients with unilateral retinoblastoma. METHODS: Samples of paraffin-embedded tumour tissue from 43 children with unilateral retinoblastoma were collected to investigate the presence of HPV DNA using polymerase chain reaction (PCR) and dot blot hybridization. RESULTS: Oncogenic HPV DNA types 16 and 35 were detected in 12 (27.9%) of 43 tumour specimens. A higher frequency of differentiated tumours (63.3%) was observed among the HPV-positive tumours. CONCLUSIONS: Future studies are necessary to demonstrate an association between HPV and sporadic retinoblastoma.

Virus Coat Protein Transgenic Papaya Provides Practical Control of Papaya ringspot virus in Hawaii.

Since 1992, Papaya ringspot virus (PRSV) destroyed nearly all of the papaya hectarage in the Puna district of Hawaii, where 95% of Hawaii's papayas are grown. Two field trials to evaluate transgenic resistance (TR) were established in Puna in October 1995. One trial included the following: SunUp, a newly named homozygous transformant of Sunset; Rainbow, a hybrid of SunUp, the nontransgenic Kapoho cultivar widely grown in Puna, and 63-1, another segregating transgenic line of Sunset. The second trial was a 0.4-ha block of Rainbow, simulating a near-commercial planting. Both trials were installed within a matrix of Sunrise, a PRSV-susceptible sibling line of Sunset. The matrix served to contain and trace pollen flow from TR plants, and as a secondary inoculum source. Virus infection was first observed 3.5 months after planting. At a year, 100% of the non-TR control and 91% of the matrix plants were infected, while PRSV infection was not observed on any of the TR plants. Fruit production data of SunUp and Rainbow show that yields were at least three times higher than the industry average, while maintaining percent soluble solids above the minimum of 11% required for commercial fruit. These data suggest that transgenic SunUp and Rainbow, homozygous and hemizygous for the coat protein transgene, respectively, offer a good solution to the PRSV problem in Hawaii.

Temporal effects of estradiol (E) on luteinizing hormone-releasing hormone (LHRH) and LH release in castrated male sheep.

We tested the hypothesis that rapidly expressed inhibitory effects of estradiol (E) on luteinizing hormone (LH) release in the male are attributable, in part, to suppression of luteinizing hormone-releasing hormone (LHRH) release. Hypophyseal-portal cannulated, castrated male sheep were infused with E (15 ng/kg/hr) or vehicle. Portal and jugular blood samples were collected at 10-min intervals for 4 hr before, and for either 12 hr (E, n = 4; vehicle, n = 4) or 24 hr (E, n = 8; vehicle, n = 3) after the start of infusion. In animals sampled for 16 hr, temporal changes in both LHRH and LH were assessed. In animals sampled for 28 hr, only LH data were analyzed. Before either the 12-hr or 24-hr infusion, LHRH and/or LH mean concentrations, pulse amplitude and interpulse interval (IPI) did not differ between E- and vehicle-infused animals. In animals sampled for 16 hr, no effects of time or steroid x time interactions were detected for mean LHRH and LHRH pulse amplitude; however, both were greater (P < 0.01) in vehicle-infused than in E-infused males. LHRH IPI was unaffected by infusion. In contrast, both mean LH and LH pulse amplitude declined (P < 0.01) within 4-8 hr after the start of E infusion, whereas mean LH IPI was unaffected. In animals sampled for 28 hr, an effect of time (P < 0.01) and a steroid x time interaction (P < 0.01) was detected for mean LH, and there was an effect of time (P < 0.01) on LH pulse amplitude. Mean LH IPI was not affected. Our results show that in male sheep E rapidly reduces LH release in the absence of a detectable change in LHRH release.

Effects of dialyzing gamma-aminobutyric acid receptor antagonists into the medial preoptic and arcuate ventromedial region on luteinizing hormone release in male sheep.

We investigated the effects of microdialyzing alpha-aminobutyric acid (GABA) receptor antagonists into either the medial preoptic area (mPOA) or the arcuate-ventromedial region (ARC-VMR) on LH secretion. Bicuculline methiodide (BMI, GABA(A) receptor antagonist), and either 2-hydroxysaclofen (SAC) or CGP 55845A (CGP, GABA(B) receptor antagonists) were used. In experiment 1, castrated rams received 4-h dialysis into either the mPOA (n = 5) or ARC-VMR (n = 4) of artificial cerebrospinal fluid (aCSF) followed by 4 h of either BMI (aCSF-BMI, 375 microM in mPOA, 1 mM in the ARC-VMR for 2-1/2 h), or aCSF-SAC (5 mM). In experiment 2, castrated rams received dialysis only in the ARC-VMR (n = 5) of aCSF-aCSF, aCSF-BMI (375 microM), or aCSF-CGP (50 microM). In experiment 3, untreated or testosterone (T)-treated castrated rams (n = 6/group) received dialysis only in the ARC-VMR of aCSF-aCSF, aCSF-BMI (375 microM), or aCSF-CGP (500 microM). Jugular blood was collected at 10-min intervals. In experiment 1, BMI suppressed mean plasma LH (p < 0.05) and increased interpulse interval (IPI, p < 0.05) at both sites. In experiment 2, BMI significantly reduced mean LH and increased IPI (p < 0.01). In experiment 3, BMI reduced mean LH in both the presence (p < 0.05) and absence of T (p < 0.01) and increased IPI (p < 0.01) in the absence of T. SAC, CGP, and aCSF did not affect LH in any experiment. These results show that dialysis of BMI, into either the mPOA or the ARC-VMR of either castrated or T-treated castrated rams decreased LH release, whereas dialysis of GABA(B) antagonists at these sites was without detectable effect.

Effect of infusing gamma-aminobutyric acid receptor agonists and antagonists into the medial preoptic area and ventromedial hypothalamus on prolactin secretion in male sheep.

We investigated the effects of gamma-aminobutyric acid (GABA) agonists muscimol and baclofen (GABA(A) and GABA(B) agonists, respectively) and antagonists bicuculline methiodide (BMI, GABA(A) antagonist) or 2-hydroxysaclofen (SAC) and CGP 55845A (GABA(B) antagonists) on prolactin (PRL) secretion in castrated rams. The drugs were applied by microdialysis into either the medial preoptic area (mPOA) or ventromedial hypothalamus (VMH). Dialysis of baclofen into the mPOA significantly increased mean PRL (p < 0.05), whereas SAC caused a small, but significant decrease (p < 0.01). Dialysis of either muscimol or BMI into the mPOA had no effect on prolactin. In the VMH, baclofen significantly increased (p < 0.01) mean PRL but SAC and CGP 55845A were ineffective, whereas dialysis of either muscimol or BMI increased mean prolactin (p < 0.01). These results show that infusion into the mPOA of drugs that affect GABA(B) receptor alter PRL release, whereas infusion of a GABA(A) agonists and antagonist was without effect on PRL release. In contrast, infusion of both GABA(A) and GABA(B) agonists and a GABA(A) antagonist into the VMH altered PRL secretion. This suggest that GABAergic neurons in both regions participate in regulating PRL secretion, but by different receptor systems.

Hypothalamic sites of action for testosterone, dihydrotestosterone, and estrogen in the regulation of luteinizing hormone secretion in male sheep.

Testosterone (T) inhibits LH secretion partly by acting at unknown sites within the brain to inhibit GnRH secretion. We tested the hypothesis that the preoptic area (POA) and arcuate-ventromedial region (ARC/VMR), areas rich in androgen and estrogen (E) receptors, are neural sites at which T and the T metabolites, dihydrotestosterone (DHT) and estrogen (E), act to suppress LH secretion. Bilateral guide cannulae were surgically implanted into either the POA or ARC/VMR of castrated male sheep. Experiments were conducted under a long day photoperiod to maximize the inhibitory effect of the steroids. In Exp 1, all sheep (n = 6/site) sequentially received bilateral implants of cholesterol (CHOL), T, or E at each site. Jugular blood samples were taken at 10-min intervals for 4 h both immediately before implant insertion and 5 days later. In Exp 2, all sheep (n = 6/site) sequentially received bilateral implants of CHOL, DHT, or E at each site according to a latin square design. Blood samples were taken before and 7 days after implant insertion. In Exp 3, which followed the same design as Exp 2, implants of E, T, or DHT were placed only in the ARC/VMR. In the final experiment, the effects of T and CHOL implants in the ARC/VMR were compared. Neither T, DHT, nor CHOL implants at either site affected LH secretion. In contrast, E treatment in the ARC/VMR suppressed mean plasma LH levels (P < 0.01), primarily due to an increase in interpulse interval (P < 0.01). Estrogen implants in the POA caused a small, but nonsignificant (P > 0.05), decrease in mean LH levels in the first experiment and an increase in LH interpulse interval (P < 0.05) in the second experiment. These results suggest that the ARC/VMR and possibly the POA are sites at which E acts to reduce GnRH secretion in male sheep.

Pharmacokinetics of extracellular melatonin in Siberian hamster forebrain.

In vivo brain microdialysis was used to characterize the pharmacokinetics of subcutaneously injected melatonin in the anterior hypothalamic-preoptic area (AH-POA) of the male Siberian hamster. Animals with a microdialysis probe implanted in the AH-POA were treated with a subcutaneous melatonin injection at 0900 h (3 h after lights-on) or 2000 h (2 h prior to lights-off). Treatment with 2.5 or 0.25 mg/kg dosages of melatonin in saline vehicle induced peak concentrations of melatonin in AH-POA microdialysates within 20 min of injection (165 +/- 34 and 18 +/- 8 pg/20 min, respectively). For the 2.5 and 0.25 mg/kg dosages, the half-life of the absorbed melatonin (t 1/2 elimination) was less than 20 min, and the concentrations fell to baseline within 60 min after injection. There were no significant time of day effects on the kinetic profile of extracellular melatonin associated with either of these dosages. These results confirm the rapid accumulation and clearance of extracellular melatonin in the vicinity of its putative target tissues.

Differential regulation of luteinizing hormone release by gamma-aminobutyric acid receptor subtypes in the arcuate-ventromedial region of the castrated ram.

We investigated the effects on LH secretion of infusing gamma-aminobutyric acid (GABA) agonists muscimol and baclofen (GABAA and GABAB receptor agonists, respectively) into either the medial preoptic area (mPOA) or the arcuate-ventromedial region (ARC-VMR) of the hypothalamus of castrated rams during the nonbreeding season. Bilateral microdialysis of artificial cerebrospinal fluid for 4 h followed by treatment with artificial cerebrospinal fluid, baclofen (1 mM), or muscimol (1 mM in the ARC-VMR, 250 microM in the mPOA) for 4 h was carried out on three separate occasions in random order. Simultaneously, jugular venous blood was collected at 10-min intervals, and plasma later was assayed for LH. The estimated dose of baclofen delivered to each unilateral microdialysis site was 7.9 micrograms; for muscimol, it was 1.1 micrograms for the mPOA and 4.5 micrograms for the ARC-VMR. In the mPOA, baclofen had no detectable effect, whereas muscimol had a delayed suppressive effect on mean LH (P < 0.01). In the ARC-VMR muscimol rapidly reduced mean LH (P < 0.01). In contrast, baclofen increased mean LH (P = 0.01) and LH pulse amplitude (P = 0.05) without altering the LH interpulse interval (P > 0.10). These results support the involvement of GABAA receptors in the mPOA in regulating LH secretory patterns. More importantly, both GABAA and GABA(B) receptors in the ARC-VMR appear to differentially modulate LH and, presumably, GnRH release. Whether GABA acts directly on the GnRH neuron or indirectly through other neural systems remains to be determined.

Diurnal variation in 5-hydroxyindole-acetic acid output in the suprachiasmatic region of the Siberian hamster assessed by in vivo microdialysis: evidence for nocturnal activation of serotonin release.

In vivo brain microdialysis was used to characterize the daily pattern of 5-hydroxyindole-acetic acid (5-HIAA) release in the region of the suprachiasmatic nuclei (SCN) in freely behaving male Siberian hamsters housed under 16L:8D. A marked diurnal variation in the concentration of extracellular 5-HIAA was apparent, with peak levels (147 +/- 5% of the daily mean; p < 0.05) occurring 2-3 h after lights-off. Smaller nocturnal rises in extracellular 5-HIAA were observed in the posterior hypothalamus and preoptic area (128 +/- 4 and 123 +/- 8% of the daily mean, respectively; both p < 0.05 vs. average daytime levels). Tryptophan loading increased 5-HIAA in SCN microdialysates by 44 +/- 6%, and this response was enhanced by localized perfusion with tetrodotoxin (TTX; 5 microM). Localized applications of KCl (150 mM) or veratridine (100 microM) decreased 5-HIAA by 62 +/- 5 or 49 +/- 11%, respectively. The effect of KCl was not significantly affected by specific calcium channel blockers. Perfusion with TTX markedly decreased SCN 5-HIAA during the dark phase, but had little effect during the light phase (42 +/- 8 vs. 12 +/- 5% suppression, respectively; p < 0.01). Addition of serotonin (3 microM) to the perfusate significantly stimulated 5-HIAA output. This treatment increased the release of 5-HIAA more during the dark than during the light phase (61 +/- 8 vs. 25 +/- 5%, respectively; p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

[Mitral valve prolapse in the 8th and 9th decades of life. Anatomo-pathologic study of 3 cases]. / Prolapso de valva mitral nas 8a. e 9a. décadas de vida. Estudo anátomo-patológico em três casos.

Different anatomo-clinic aspects from three mitral valve prolapse cases are compared to those commonly presented in the literature and are also utilized as a basis for a new classification of this disease. The patients are more than 77 years old, what is in contrast with the current concept of MVP as a disease of young-middle aged women. The first case shows marked ostial dilation and many ruptured chordae: as a consequence, this patient showed severe cardiac dysfunction. The anterior, rather than the posterior leaflet, presented intense myxoid degeneration. In the second case, no ruptured chordae were detected and, consequently, the degree of heart failure was lesser than the first one, in spite of the same degree of ostial dilation. Both leaflets showed the same degree of myxoid degeneration. The third patient, who does not have heart failure, showed myxomatous degeneration of both cusps, but no ostial dilation or chordal rupture were present. These aspects reinforce the impression that isolated mixomatous degeneration of the cusps is not so deleterious when compared to those cases where the mitral valvar ring is dilated or its chordae are also involved by that degenerative process. Therefore ostial dimension (normal or enlarged) and the state of the chordae (with or without rupture) appears to be important points to be considered in MVP.