Vascular endothelial growth factor (VEGF)-D deficiency had no significant effect on total body weight or on subcutaneous (SC) or gonadal (GON) adipose tissue mass of mice kept on a standard fat (SFD) or a high fat diet (HFD) for 15 weeks. The composition of SC and GON adipose tissues of VEGF-D deficient mice in terms of size and density of adipocytes or blood vessels was also comparable to that of wild-type control mice. Staining of lymphatic vessels in adipose tissue sections did not reveal marked differences between both genotypes. The absence of an effect of VEGF-D deficiency could not be explained by compensatory increases of VEGF-C expression in adipose tissues of the deficient mice. Thus, our data do not support an important role of VEGF-D in (lymph) angiogenesis or in adipose tissue development.

BACKGROUND

The Breathmobile, a specialty-based mobile asthma clinic, provides free care to underserved children. The cost of symptom-free day (SFD) improvement in this population has not been previously reported.

OBJECTIVE

To examine the clinical impact and cost-effectiveness of the Baltimore Breathmobile.

METHODS

Existing computerized data were analyzed for Breathmobile patient visits between 2002 and 2007. All SFDs were calculated, and direct medical cost savings attributable to decreased emergency department visits and hospitalizations (after program utilization vs the previous year) were compared with annual operating costs. Incremental cost-effectiveness ratios were determined by calculating the incremental costs of Breathmobile care per additional SFD gained per child per year.

RESULTS

The analysis included 255 patients enrolled in the program for at least 1 year. Most participants were black (93.3%), and 54.9% were male. At baseline, patients reported a mean (SD) of 199 (118) SFDs in the year before enrollment. After 1 year in the program, patients had a mean (SD) improvement of 44 (9) SFDs. The program resulted in overall cost savings of $79.43 per SFD gained, with greater cost savings for children aged 5 to 11 years (-$116.84 per SFD gained) and those with intermittent asthma (-$126.71 per SFD gained).

CONCLUSIONS

The Baltimore Breathmobile program has demonstrated significant improvement in SFDs, with direct medical cost savings of the program outweighing the operational costs. These data support the need to continue to sustain and expand Breathmobile programs for children at high risk for asthma exacerbations and to conduct a randomized clinical trial to estimate the cost-effectiveness of the Breathmobile.

OBJECTIVE

Test-retest reliability of an asthma paper diary versus an electronic diary (e-diary) with an integrated peak flow meter was investigated. The equivalence of the two modes was also evaluated.

METHODS

Prospective, randomized crossover study design in adolescents (12-17 years) and adults (≥18 years). Key inclusion criteria were persistent asthma, Asthma Control Test (ACT) scores ≥16, use of inhaled corticosteroid with or without long-acting beta-agonist for ≥12 weeks, nocturnal awakenings <2 times in the past week, and activity limitations <1 per week. Participants were randomized to either paper then e-diary or e-diary then paper, to be completed for 14 days each.

RESULTS

Forty-seven participants completed all study visits. Weekly percentage of symptom-free days (SFDs) and rescue-free days (RFDs) were calculated. Intraclass correlation coefficients (ICCs) of Week 1 mean SFD and RFD (test) and Week 2 mean SFD and RFD (retest), respectively, were estimated in three groups defined as stable: (i) minimal changes in asthma symptoms, as measured by the global patient reported symptom change question, (ii) less than 15% change (absolute value) in 1 second FEV(1) at adjacent study visits, and (iii) changes in ACT scores less than three points for each mode. SFD demonstrated acceptable ICC (≥0.70) using all three definitions of asthma stability for both modes.

CONCLUSIONS

Acceptable reproducibility of the percentage of RFD (ICC = 0.78) was only observed for the e-diary using the FEV(1) stability criterion. The ICCs for SFD and RFD were acceptable, 0.84 and 0.70, respectively, suggesting better reliability for the e-diary.

The purpose of this study was to perform comprehensive measurements and testing of a Novalis Tx linear accelerator, and to develop technical guidelines for com-missioning from the time of acceptance testing to the first clinical treatment. The Novalis Tx (NTX) linear accelerator is equipped with, among other features, a high-definition MLC (HD120 MLC) with 2.5 mm central leaves, a 6D robotic couch, an optical guidance positioning system, as well as X-ray-based image guidance tools to provide high accuracy radiation delivery for stereotactic radiosurgery and stereotactic body radiation therapy procedures. We have performed extensive tests for each of the components, and analyzed the clinical data collected in our clinic. We present technical guidelines in this report focusing on methods for: (1) efficient and accurate beam data collection for commissioning treatment planning systems, including small field output measurements conducted using a wide range of detectors; (2) commissioning tests for the HD120 MLC; (3) data collection for the baseline characteristics of the on-board imager (OBI) and ExacTrac X-ray (ETX) image guidance systems in conjunction with the 6D robotic couch; and (4) end-to-end testing of the entire clinical process. Established from our clinical experience thus far, recommendations are provided for accurate and efficient use of the OBI and ETX localization systems for intra- and extracranial treatment sites. Four results are presented. (1) Basic beam data measurements: Our measurements confirmed the necessity of using small detectors for small fields. Total scatter factors varied significantly (30% to approximately 62%) for small field measurements among detectors. Unshielded stereotactic field diode (SFD) overestimated dose by ~ 2% for large field sizes. Ion chambers with active diameters of 6 mm suffered from significant volume averaging. The sharpest profile penumbra was observed for the SFD because of its small active diameter (0.6 mm). (2) MLC commissioning: Winston Lutz test, light/radiation field congruence, and Picket Fence tests were performed and were within criteria established by the relevant task group reports. The measured mean MLC transmission and dynamic leaf gap of 6 MV SRS beam were 1.17% and 0.36 mm, respectively. (3) Baseline characteristics of OBI and ETX: The isocenter localization errors in the left/right, posterior/anterior, and superior/inferior directions were, respectively, -0.2 ± 0.2 mm, -0.8 ± 0.2 mm, and -0.8 ± 0.4 mm for ETX, and 0.5 ± 0.7 mm, 0.6 ± 0.5 mm, and 0.0 ± 0.5 mm for OBI cone-beam computed tomography. The registration angular discrepancy was 0.1 ± 0.2°, and the maximum robotic couch error was 0.2°. (4) End-to-end tests: The measured isocenter dose differences from the planned values were 0.8% and 0.4%, measured respectively by an ion chamber and film. The gamma pass rate, measured by EBT2 film, was 95% (3% DD and 1 mm DTA). Through a systematic series of quantitative commissioning experiments and end-to-end tests and our initial clinical experience, described in this report, we demonstrate that the NTX is a robust system, with the image guidance and MLC requirements to treat a wide variety of sites - in particular for highly accurate delivery of SRS and SBRT-based treatments.

In aquaculture, disinfection of facilities, prevention of fish diseases, and stimulation of fish growth are priority goals and the most important sources of toxic substances to the environment, together with excretory products from fish. In the present study, embryos of two species of sea urchin (Paracentrotus lividus and Arbacia lixula) were exposed to serial dilutions of six antibiotics (amoxicillin (AMOX), ampicillin, flumequine (FLU), oxytetracycline (OTC), streptomycin (ST), and sulfadiazine [SFD]) and two disinfectants (sodium hypochlorite (NaClO) and formaldehyde [CH(2)O]). Alterations in larval development were studied, and the effective concentrations (ECs) were calculated to evaluate the toxicity of the substances. Both species showed similar sensitivities to all substances tested. Disinfectants (EC(50) = 1.78 and 1.79 mg/l for CH(2)O; EC(50) = 10.15 and 11.1 mg/l for NaClO) were found to be more toxic than antibiotics. AMOX, OTC, and ST caused <20 % of alterations, even at the highest concentrations tested. FLU was the most toxic to P. lividus (EC(50) = 31.0 mg/l) and SFD to A. lixula (EC(50) = 12.7 mg/l). The sea urchin bioassay should be considered within toxicity assessment-monitoring plans because of the sensitivity of larvae to disinfectants.

Sandy grassland restoration is a vital process including re-structure of soils, restoration of vegetation, and soil functioning in arid and semi-arid regions. Soil fungal community is a complex and critical component of soil functioning and ecological balance due to its roles in organic matter decomposition and nutrient cycling following sandy grassland restoration. In this study, soil fungal community and its relationship with environmental factors were examined along a habitat gradient of sandy grassland restoration: mobile dunes (MD), semi-fixed dunes (SFD), fixed dunes (FD), and grassland (G). It was found that species abundance, richness, and diversity of fungal community increased along with the sandy grassland restoration. The sequences analysis suggested that most of the fungal species (68.4 %) belonged to the phylum of Ascomycota. The three predominant fungal species were Pleospora herbarum, Wickerhamomyces anomalus, and Deconica Montana, accounting for more than one fourth of all the 38 species. Geranomyces variabilis was the subdominant species in MD, Pseudogymnoascus destructans and Mortierella alpine were the subdominant species in SFD, and P. destructans and Fungi incertae sedis were the dominant species in FD and G. The result from redundancy analysis (RDA) and stepwise regression analysis indicated that the vegetation characteristics and soil properties explain a significant proportion of the variation in the fungal community, and aboveground biomass and C:N ratio are the key factors to determine soil fungal community composition during sandy grassland restoration. It was suggested that the restoration of sandy grassland combined with vegetation and soil properties improved the soil fungal diversity. Also, the dominant species was found to be alternative following the restoration of sandy grassland ecosystems.

OBJECTIVE

There is scarcity of information on impact of iodine deficiency on growth and development of newborns, both pre-term and term babies. The present study was carried out to see the impact of iodine deficiency (ID) on the intrauterine foetal growth and development in terms of birth weight (BW) in an African population living in an iodine deficient zone.

METHODS

The maternal and cord serum thyroid parameters at term delivery (38-40 wk of gestation) were measured and correlated with the birth weights of the corresponding newborns, and the results compared between those assessed in an ID zone (Jengere region of Bassa district, Plateau State, Nigeria) and with that of non ID (Control) zone, Jos, the State capital.

RESULTS

The level of maternal and neonatal thyroid function were significantly reduced in the group with low birth weights (<2.5 kg) as compared to the level seen in normal birth weights category in both control (Jos) and ID (Jengere) regions. Both in non ID and ID regions the maternal serum thyroxine (T4), thyroxine binding globulin (TBG) and free thyroxine index (FT4I) were positively correlated (P<0.005) with BW of term babies while thyroid stimulating hormone (TSH) (P<0.005) and thyroxine binding capacity (TBK) (P<0.05) showed negative correlation. Maternal T3 and T3/T4 ratio did not show any correlation with BW in Jos, while in the ID region of Bassa the BW showed a positive correlation (P<0.005) with the maternal serum T3 and T3/T4 ratio. Cord serum analysis of the term babies revealed that the BW was positively correlated with its T4, triiodothyronine (T3), TBG, FT4I and T4/TBG ratio (Jos only) (P<0.005) in both ID region of Bassa and non ID control zone of Jos. Cord serum TSH and TBK showed negative correlation (P<0.005) with BW. Thyrotrophin releasing hormone (TRH) stimulation test revealed that mothers with small for date (SFD) babies at term were more hypothyroid compared to the level of thyroid function seen in the women delivering normal babies.

CONCLUSIONS

Maternal thyroid hormone plays a significant role in the intrauterine foetal growth and development, and the smaller babies and their corresponding mothers, in an ID affected area are functionally sub-thyroid in greater proportion due to prevailing long standing environmental iodine deficiency (EID). Maternal T3 is an important factor in the defence of foetus in ID regions.

Right ventricular (RV) function is a strong predictor of outcome in cardiovascular diseases. Two components of RV function, longitudinal and transverse motion, have been investigated in pulmonary hypertension (PH). However, their individual clinical significance remains uncertain. The aim of this study was to determine the factors associated with transverse and longitudinal RV motion in patients with PH. In 149 treatment-naive patients with PH and 16 patients with suspected PH found to have mean pulmonary arterial pressure of <20 mmHg, cardiovascular magnetic resonance imaging was performed within 24 hours of right heart catheterization. In patients with PH, fractional longitudinal motion (fractional tricuspid annulus to apex distance [f-TAAD]) was significantly greater than fractional transverse motion (fractional septum to free wall distance [f-SFD]; P = 0.002). In patients without PH, no significant difference between f-SFD and f-TAAD was identified (P = 0.442). Longitudinal RV motion was singularly associated with RV ejection fraction independent of age, invasive hemodynamics, and cardiac magnetic resonance measurements (P = 0.024). In contrast, transverse RV motion was independently associated with left ventricular eccentricity (P = 0.036) in addition to RV ejection fraction (P = 0.014). In conclusion, RV motion is significantly greater in the longitudinal direction in patients with PH, whereas patients without PH have equal contributions of transverse and longitudinal motion. Longitudinal RV motion is primarily associated with global RV pump function in PH. Transverse RV motion not only reflects global pump function but is independently influenced by ventricular interaction in patients with PH.

A blood blister-like (BBL) or dissecting aneurysm should be carefully considered if located at a non-branching site of the supra-clinoid internal carotid artery (ICA). Several surgical and endovascular treatment methods have been proposed but they all carry a relatively high risk of morbidity and mortality. This study evaluated the effectiveness of a novel Silk flow-diverting device (SFD) placed in the early acute stage. Three patients presenting with acute subarachnoid haemorrhage caused by small blister-like aneurysms of the carotid siphon were treated within 48 hours after admission by placement of SFDs. More than one device was placed to cover the lesion. None of the patients were premedicated and started anti-platelet therapy during the procedure. All aneurysms were successfully occluded. A good outcome was observed in two out of three treated patients. No thromboembolic or haemorrhagic event occurred during or after the procedures, or during follow-up (6-14 months). SFD prevented rebleeding and the use of these devices could be proposed as an option to treat fragile uncoilable BBL aneurysms, even in the early acute phase without anti-platelet premedication. Larger studies and long-terms results are necessary.

OBJECTIVE

A case of choroidal neovascularization (CNV) secondary to Sorsby fundus dystrophy (SFD) treated with systemic bevacizumab (Avastin).

METHODS

A 41-year-old woman presented with CNV secondary to SFD in her better eye. The patient received three initial infusions of bevacizumab at a dose of 5 mg/kg at 2 week intervals and one additional infusion because of CNV recurrence at the 7 month follow-up.

RESULTS

At 16 month follow-up, visual acuity had improved from 20/50 at baseline to 20/25; optical coherence tomography and fluorescein angiography showed no evidence of CNV activity.

CONCLUSIONS

Systemic bevacizumab seems to be a promising treatment option for CNV secondary to SFD.

BACKGROUND

The DSM-5 working group on the somatoform (SFD) and factitious (FD) disorders has recommended substantial revisions of these categories. The recommendations are based, in part, on anecdotal evidence that the diagnoses are infrequently used.

OBJECTIVE

To assess the assignment rates for SFD, FD, and related diagnoses among general medical inpatients.

METHODS

The National Hospital Discharge Survey was queried for instances of SFD and FD, along with related diagnoses identifying medical cases in which psychological factors play a role. Diagnoses of major depression and generalized anxiety disorder were queried for comparison purposes.

RESULTS

The target diagnoses were assigned far less frequently than published prevalence and recognition rates suggest. Nearly half of the assigned target diagnoses were generic diagnoses (esp. physiological malfunction due to psychological factors) other than SFD or FD. However, the apparent degree of underassignment of the target diagnoses was not dramatically greater than the underassignment observed for major depression and generalized anxiety disorder.

CONCLUSIONS

The results provide empirical support for the impression that physicians do not assign SFD and FD diagnoses in recognized cases, but do not strongly support the assertion that these diagnoses are uniquely problematic.

OBJECTIVE

To determine whether musculoskeletal pain was associated with impaired sexual function in a population sample of middle-aged and older men.

METHODS

The European Male Ageing Study (EMAS), a multicenter population-based study of men aged 40-79 years, was used to investigate this hypothesis. A questionnaire asked about the presence and duration of musculoskeletal pain, allowing subjects to be classified into 1 of 3 groups: those reporting chronic widespread pain (CWP), those reporting pain but not CWP ("some pain"), and those with no pain. Subjects completed a sexual function questionnaire from which 3 domains were considered: overall sexual functioning (OSF), sexual functioning-related distress (SFD), and change in sexual functioning compared to 1 year ago (CSF).

RESULTS

A total of 3206 men [mean age 60 (SD 11) yrs] had complete data on pain status. Of these, 8.7% had CWP and 50.34% had "some pain." Pain was associated with lower OSF, and higher SFD and CSF scores. After adjustment for putative confounding factors, the associations became non-significant with OSF and CSF but persisted for SFD. Associations between pain status and some items within the sexual functioning domains, including frequency of sexual intercourse, frequency of morning erections, sexual desire, and orgasm were also significant, although these associations varied by pain status.

CONCLUSIONS

Musculoskeletal pain is associated with several aspects of sexual functioning. These relationships differ depending on the extent of the pain (chronic or not) and are also largely confounded by other health-related factors, primarily depression.

A comparative study of patients with premature thelarche and patients with idiopathic true precocious puberty was conducted. The age at the first visit tended to be lower for those with precocious puberty. In comparison with normative data for children, the frequency of low birthweight and small for date (SFD) status was greater in the 55 patients with premature thelarche, but SFD was also frequent in the 18 patients with precocious puberty. The height, weight and Kaup's index were all within the normal range for these two groups. The ratios for bone age/chronologic age and bone age/height age tended to be high in both groups. In the patients with premature thelarche, the blood luteinizing hormone (LH) level showed a normal response, and the blood follicle stimulating hormone (FSH) level a hyper-response, to stimulation with luteinizing hormone-releasing hormone (LH-RH). In contrast, both the blood LH and FSH levels showed a normal response to LH-RH stimulation in most of the patients with precocious puberty, and a hyper-response was rare among them. Although the blood estradiol (E2) level was higher in patients with precocious puberty than in those with premature thelarche, about 50% and 90% of the patients in the respective groups had normal levels. These results suggest that normal responses of blood LH and excessive responses of blood FSH to LH-RH loading may be useful in some patients for diagnosing premature thelarche at an early stage.

BACKGROUND

In 2003, an ICRC-SFD Ponseti program was introduced in southern Vietnam. Additional programs were introduced by the Prosthetics Outreach Foundation and independently by physicians trained at our center. The purpose of this study was to evaluate the impact, progress and challenges facing Ponseti practitioners and patients' family members in Vietnam. In addition, web-conferencing (Ponseti Virtual Forum) for continued medical education in the method was also assessed.

METHODS

Multiple questionnaires were developed to conduct face-to-face practitioner interviews, focus group interviews, and parental interviews. Observation was done at multiple site clinics to determine or confirm additional challenges faced by practitioners. Web conferencing was introduced to sites in Ho Chi Minh City and Da Nang City.

RESULTS

The number of clubfoot patients treated with the Ponseti method has increased over time with approximately 1,252 infants treated between 2003 and 2010. Specific challenges were identified relating to communication, networking, distance and transportation, and finances for both practitioners and parents. The PVF was not only found to facilitate rapid, relevant dissemination of medical knowledge--thus increasing physician and patient satisfaction--but it may also be found to act as an interface in which medical culture, insight, and compassion are shared benefiting all virtual forum participants.

CONCLUSIONS

The identified progress and challenges mirrored that of similar studies done in other countries with several factors affecting progress. Focusing on improving communication channels and networking while working with the ministry of health may improve the facilitation of the Ponseti method in Vietnam. Further implementation and evaluation of the PVF may act as a guide for current and future programs in Vietnam or other countries.

Dome-shaped macula (DSM) was described by Gaucher et al as a convex protrusion of macula within a staphyloma in highly myopic eyes that causes visual impairment associated with serous foveal detachment (SFD). We describe a patient with spontaneous resolution of SFD in DSM documented by serial spectral domain optical coherence tomography (SD-OCT). A 28-year-old female Japanese patient presented with blurred vision in both eyes. Upon examination, her best-corrected visual acuity was 1.2 with -8.0 sphere and 0.5 with -8.0 sphere in the right and left eyes, respectively. SD-OCT in both eyes showed an anterior bulge of the macula with SFD. The diagnosis of DSM with SFD was made on the basis of characteristic findings. Three months later, the patient achieved spontaneous resolution of SFD without treatment. One month later, there was an occurrence of SFD in the right eye, but the SFD had disappeared in 1 month. In our case, her visual impairment was proven to be a consequence of SFD in relation to DSM. SD-OCT was useful for the detection and follow-up of SFD in DSM.

A survey of the smoking habits of 3,025 couples, whose infants were receiving medical examinations, was conducted before and after pregnancy to determine the effect of smoking patterns of the parents on the newborn infant. Of the husbands 55.2% were smoking before pregnancy; 53.4% during pregnancy; and 51.8% after delivery. During pregnancy proportion of smokers decreased by only 1.8%. The percentages for wives, on the other hand, were 13.9%, 4.4%, and 5.3% respectively showing a reduction of 9.5% during pregnancy. Young couples, under 24 years of age, those with low educational backgrounds, and self-employed husbands showed the highest proportion of smokers. Examination of the relation between parents' smoking habits and their effect on the newborn infants, showed no difference in weeks of gestation nor in height. However, the infants of smoking parents were on the average 99.6 grams lighter at birth (p less than 0.02) and the rate of SFD was higher (p less than 0.05). The relative risk for SFD appearance was 1.21 with a smoking father, and 1.39 with both parents smoking. Husbands who decreased or stopped smoking during the pregnancy were largely in the 30 to 34 year old category, university graduates, and those whose wife was having their first baby. A remarkable decrease was observed in the average weight at birth of infants with a mother smoking 6 cigarettes a day or with father smoking 20 cigarettes a day compared to non-smoking parents, especially in the case of female infants.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiological studies suggest that infants of low birth weight show poor neonatal growth and increased susceptibility to adult diseases such as diabetes and lung disease. Uncoupling protein 2 and 3 (UCP2 and UCP3) have been implicated in the development of such diseases; pigs provide an ideal model to examine the influence of birth weight due to the natural variance in piglet weight within a litter. This study examined whether birth weight influences the expression of UCP2 and UCP3 in adipose tissue, skeletal muscle, and lung. Piglets from 11 litters were ranked according to birth weight and three from each litter assigned to small (SFD), normal (NFD), or large for dates (LFD) groups. Blood samples and morphometric measurements were taken over the first 14 days of life, and tissue samples were taken on day 7 or 14. Plasma hormone and metabolite concentrations and the expression of UCP2 and UCP3 mRNA in adipose tissue, skeletal muscle, and lung were measured. UCP2 and UCP3 expression in adipose tissue was lower in the SFD compared with the LFD group on day 7. UCP3 expression in skeletal muscle was higher than that of adipose tissue. Lung UCP2 and skeletal muscle UCP3 mRNA expression were unaffected by size at birth. Regression analysis indicated that UCP3 expression was differentially associated with IGF-1, leptin, and insulin. In conclusion, low birth weight is associated with tissue-specific effects on UCP expression. It remains to be established whether these subsequently contribute to pathological conditions such as diabetes.

Sorsby fundus dystrophy (SFD) is an autosomal dominant macular degeneration of late onset. A key feature of the disease is the thickening of Bruch's membrane, an ECM structure located between the RPE and the choroid. SFD is caused by mutations in the gene encoding the ECM-associated tissue inhibitor of metalloproteases-3 (TIMP3). We have recently generated two Timp3 gene-targeted mouse lines, one deficient for the murine gene (Timp3-/-) and one carrying an SFD-related S156C mutation. Based on extracts and cell cultures derived from tissues of these animals we now evaluated TIMP3 functionality and its contribution to SFD. We show that the activity levels of TIMP3 target proteases including TACE, ADAMTS4/5 and aggrecan-cleaving MMPs are similar in Timp3S156/+ and Timp3S156C/S156C mice when compared to controls. In Timp3-/- mice, a significant enhancement of enzyme activity was observed for TACE but not for ADAMTS4/5 and MMPs indicating a compensatory effect of other inhibitors regulating the latter two groups of proteases. Fibrin bead assays show that angiogenesis in Timp3S156/+ and Timp3S156C/S156C mice is not altered whereas increased formation of capillary tubes was observed in Timp3-/- animals over controls. Rescue experiments using recombinant proteins demonstrate that the inhibitory activities of TIMP3 towards TACE and aggrecan-cleaving MMPs as well as the anti-angiogenic properties of TIMP3 are not impaired by SFD mutation S156C. We finally demonstrate that wild-type and S156C-TIMP3 proteins block the binding of VEGF to its receptor VEGFR2 to a similar extent. Taken together, this study shows that S156C-TIMP3 retains its known functional properties suggesting that causes other than an imbalance in protease or angiogenic activities represent the primary molecular defect underlying SFD.

In the present study we report results concerning 5-hydroxytryptamine (5-HT) metabolism in two groups of small for date (SFD) human babies (gestational age 36 and 3 weeks), who suffered intrauterine nutritional restriction. A complementary study in the brain of rat fetuses with two types of intrauterine deprivation, in which brain L-tryptophan (L-Trp), tryptophan-5-hydroxylase (T5-H) activity and 5-HT content were determined on days 17, 19 and 21 of gestation. The same parameters studied prenatally were followed in both species during the immediate postnatal period. In the SFD babies the results were: (a) the free fraction of plasma L-Trp was significantly elevated; (b) plasma neutral amino acids were not substantially modified; (c) the bound fraction of L-Trp and plasma proteins were significantly low, as compared to controls. In the fetal brain of intrauterine malnourished rats, L-Trp, activity of T5-H and 5-HT content, were significantly elevated, since day 17, as related to normal littermates. These alterations in 5-HT metabolism persisted during the early postnatal period in both species. Elevation of the free fraction of plasma L-Trp in early malnourished SFD human babies suggest an increased transport of this amino acid to the brain with a possible enhancement of serotonin synthesis, during a critical period of brain differentiation.

The tissue inhibitor of metalloproteinases-3 (TIMP3) is a multifunctional protein tightly associated with the extracellular matrix (ECM). A specific type of mutation in TIMP3 which results in potentially unpaired cysteine residues at the C-terminus of the protein has been shown to cause Sorsby fundus dystrophy (SFD), an autosomal dominant retinopathy of late onset. An early finding in SFD is a striking accumulation of protein and lipid material in Bruch's membrane, a multilayered ECM structure located between the choroid and the RPE. To study the molecular mechanisms underlying SFD pathology, we recently generated two mouse lines, one deficient in Timp3 (Timp3(-/-)) and one carrying an SFD-related mutation in the orthologous murine Timp3 gene (Timp3(S156C/S156C)). We now established immortalized fibroblast cells from the mutant mouse strains and provide evidence that the various cell lines display distinct morphological and physiological features that are dependent on the mutational status of the Timp3 protein in the secreted ECM. We show that matrix metalloproteinase (MMP) activity and inhibitory properties of Timp3 are not affected by the SFD-associated mutation. We further demonstrate that Timp3(S156C) protein accumulates in the ECM of the mutant fibroblast cells and that this accumulation is not due to a prolonged turnover rate of mutant vs. normal Timp3. We also show that the relative abundance of mutant and normal Timp3 in the ECM has no measurable effects on cellular phenotypes. Together, these findings suggest (i) a functional role of normal Timp3 in pathways determining cellular morphology and (ii) a loss of this particular function as a consequence of the Ser156Cys mutation. We therefore hypothesize that SFD pathogenesis is due to a loss-of-function mutation in TIMP3.

The vacuolar proton pump of clathrin-coated vesicles is composed of two general sectors, a cytosolic, ATP hydrolytic domain (V1) and an intramembranous proton channel, V0. V1 is comprised of 8-9 subunits including polypeptides of 50 and 57 kDa, termed SFD (Sub Fifty-eight-kDa Doublet). Although SFD is essential to the activation of ATPase and proton pumping activities catalyzed by holoenzyme, its constituent polypeptides have not been separated to determine their respective roles in ATPase functions. Recent molecular characterization of these subunits revealed that they are isoforms that arise through an alternative splicing mechanism (Zhou, Z., Peng, S.-B., Crider, B.P., Slaughter, C., Xie, X.S., and Stone, D.K. (1998) J. Biol. Chem. 273, 5878-5884). To determine the functional characteristics of the 57-kDa (SFDalpha)1 and 50-kDa (SFDbeta) isoforms, we expressed these proteins in Escherichia coli. We determined that purified recombinant proteins, rSFDalpha and rSFDbeta, when reassembled with SFD-depleted holoenzyme, are functionally interchangeable in restoration of ATPase and proton pumping activities. In addition, we determined that the V-pump of chromaffin granules has only the SFDalpha isoform in its native state and that rSFDalpha and rSFDbeta are equally effective in restoring ATPase and proton pumping activities to SFD-depleted enzyme. Finally, we found that SFDalpha and SFDbeta structurally interact not only with V1, but also withV0, indicating that these activator subunits may play both structural and functional roles in coupling ATP hydrolysis to proton flow.

Shenfu decoction (SFD) can be used to treat patients with sign of Yangqi decline or Yang exhaustion related to chronic heart failure (CHF). We conducted a gas chromatography with time-of-flight mass spectrometer (GC/TOF-MS)-based metabolomic study to increase the understanding of CHF and assess the efficacies and mechanisms of SFD in treating CHF induced by coronary artery ligation in rats. Based on unsupervised principal component analysis, there was a clear separation between the CHF and sham surgery group, which revealed that CHF disturbed the metabolism of endogenous substances and significantly altered the urine metabolite fingerprints. After SFD treatment, the metabolomics profile found in CHF was significantly reversed, shifting much closer to normal controls and sham surgery group, indicating that SFD has therapeutic effects in CHF, which is in accordance with the hemodynamic assay results. Metabolomic pathway analysis demonstrated that several pathways including fatty acid biosynthesis, fatty acid elongation, steroid biosynthesis, galactose metabolism, and amino acid metabolism were significantly altered in CHF rats. Therefore, we may infer that SFD shows therapeutic efficacy in CHF by restoring these disturbed metabolic pathways, especially those related to energy metabolism. This study offers new methodologies for increasing the understanding of CHF and systematically characterizing the efficacies and mechanisms of SFD in treating CHF.

This experiment was designed to study in rats the implications of the dietary type of fat at two levels of vitamin E on the life span as well as on several biochemical and anatomopathological age-related changes. For this purpose, six different isoenergetic diets containing 15% coconut oil (SFD), safflower oil (UFD) or a combination of both (CFD) with 2 or 200 mg% of dl-alpha-tocopherol were offered ad libitum to outbred Wistar male rats from weaning to senescence. The results indicated that up to 9--12 months the body weights of rats consuming the CFD or the UFD increased generally faster than those fed the SFD, and that all rats developed moderate degrees of obesity. Age-dependent changes in organ weights (kidneys, testes, spleen, brain, liver and heart) were unaffected by diet. Serum levels of vitamin E generally reflected the corresponding dietary levels, but were also influenced by the type of dietary fat. Serum cholesterol levels were not significantly affected by the type of diet or by age. Only transient hypotriglyceridemic and hypophospholipidemic effects of the UFD were observed and, while the levels of triglycerides decreased with age up to the 18th month followed by an increase at 24 months, the levels of serum phospholipids remained unchanged. Neither diet nor age modified the serum albumin/globulin ratios. While no differences in maximum life span were found between dietary groups, the 50% survival time of rats fed the UFD at high level of vitamin E was significantly longer than in all the other groups. This beneficial effect was related to postponement of the onset and reduction of incidence of malignant neoplasms, but was apparently not related to any particular influence on the incidence or severity of chronic nephropathy which practically developed in all rats. Various neoplastic, degenerative and inflammatory diseases encountered in rats dying during the course of the experiment were tabulated and compared with similar findings reported by others in different strains of rats. Pituitary and adrenocortical adenomas as well as adrenocortical and renal carcinomas were the most frequent tumors found in this study. All the pathological changes provided useful baseline information for the evaluation of data presented in this and subsequent communications of this series of studies.

The gene corticotropin releasing hormone (CRH) is mapped on bovine chromosome 14 (BTA14), where more than 30 fat-related quantitative trait loci (QTLs) have been reported in dairy and beef cattle. The gene product regulates secretion of adrenocorticotrophin hormone, the hypothalamic-pituitary-adrenal axis, and multiple hypothalamic functions; therefore, we hypothesized that CRH is a promising candidate gene for beef marbling score (BMS) and subcutaneous fat depth (SFD) in a Wagyu x Limousin F2 population. Two pairs of primers were designed and a total of 5 single nucleotide polymorphisms (SNPs) were identified: g.9657C>T, c.10718G>C, c.10841G>A, c.10893A>C, and c.10936G>C (AAFC03076794.1). Among the 4 cSNPs, c.10718G>C, c.10841G>A, and c.10936G>C are missense mutations leading to amino acid changes from arginine to proline, from serine to asparagine, and from aspartic acid to histidine, respectively. These 5 SNPs were genotyped on ~250 F2 progeny, but only 4 were selected as tagging SNPs for association analysis because no historical recombination was observed between c.10718G>C and c.10893A>C. Statistical analysis showed that g.9657C>T, c.10718G>C, and c.10936G>C and their haplotypes had significant effects on SFD, but only c.10936G>C was significantly associated with BMS. The SNP in the promoter (g.9657C>T) led to gain/loss of a CpG site and 4 potential regulatory binding sites. Different haplotypes among the 4 cSNPs significantly affected mRNA secondary structures but were not associated with phenotypes. Overall, our results provide further evidence that CRH is a promising candidate gene for a concordant QTL related to lipid metabolism in mammals.