French maritime pine bark extract (pycnogenol) in association with triple combination cream for the treatment of facial melasma in women: a double-blind, randomized, placebo-controlled trial.

BACKGROUND: Melasma can be recalcitrant to treatment, and relapses are common. Pycnogenol has been reported to be effective in treating melasma. OBJECTIVE: To compare the efficacy, safety and tolerability of 75 mg pycnogenol taken orally twice a day vs. a placebo, in association with the triple combination and broad-spectrum sunscreen for the treatment of facial melasma. METHODS: A randomized, double-blind, parallel, placebo-controlled study was conducted on 44 women with facial melasma in a single centre from May 2019 through November 2019. Patients with melasma were randomly assigned to orally take 75 mg pycnogenol (PYC) or a placebo (PLAC) twice a day for 60 days. Both groups also received tinted sunscreen [Sun Protection Factor (SPF) 50; Persistent Pigment Darkening (PPD) 17] for daytime use and a topical triple combination at bedtime. The primary outcome was a change from the baseline Modified Melasma Area Severity Index (mMASI) score. Secondary outcomes were improvements in the patients' quality of life (MELASQoL), colorimetric indices and Global Aesthetic Improvement Scale (GAIS). RESULTS: All participants completed the trial. The mean (SD) age of the participants was 39 (7) years, and 91% were phototypes III-IV. Both groups exhibited a reduction in mMASI scores, MELASQoL scores and colour contrast (P < 0.01). The mean (CI 95%) reductions of the mMASI scores were 49% (36-61%) for PYC and 34% (16-47%) for PLAC. The reductions in mMASI scores and colorimetric contrast were superior for the PYC group (P < 0.05). The analysis of GAIS resulted in an improvement of 86% (CI 95%: 68-96%) for the participants in the PYC group and 55% (CI 95%: 32-73%) for those in the PLAC group. There were no adverse effects related to oral treatment. CONCLUSION: Pycnogenol is well-tolerated and increases the effectiveness of broad-spectrum sunscreen and the triple combination in the treatment of facial melasma in women.

Efficacy and safety of topical isobutylamido thiazolyl resorcinol (Thiamidol) vs. 4% hydroquinone cream for facial melasma: an evaluator-blinded, randomized controlled trial.

BACKGROUND: Melasma can be refractory to treatment, and relapses are frequent. Thiamidol is a new potent tyrosinase inhibitor that has been found effective as a cosmeceutical for the depigmenting of melasma. OBJECTIVE: This study compared the efficacy and tolerability of topical 0.2% Thiamidol vs. 4% hydroquinone for facial melasma. METHODS: Fifty women with facial melasma participated in a randomized, evaluator-blinded, controlled study from September through November 2020. Patients were randomly assigned to apply a double layer of 0.2% Thiamidol twice a day or 4% hydroquinone cream at bedtime, for 90 days. Both groups received tinted sunscreen (sun protection factor 60, PPD 20). The primary outcome was the change from the baseline Modified Melasma Area Seve:rity Index (mMASI) score. Secondary outcomes were improvements in the patients' quality of life [Melasma Quality of Life Index (MELASQoL)], colourimetry, and Global Aesthetic Improvement Scale (GAIS) evaluation. RESULTS: One participant, from the hydroquinone group, did not complete the study (unrelated to adverse effects). The mean (SD) age of the participants was 43 (6) years, and 86% were phototypes III-IV. Both groups exhibited a reduction in mMASI, MELASQoL, and colour contrast scores (P < 0.01). The mean [95% confidence interval (CI 95%)] reductions of the mMASI scores were 43% (35-50%) for Thiamidol and 33% (23-42%) for hydroquinone. There was no difference between the groups in the reductions in mMASI, MELASQoL, colourimetric contrast and GAIS scores (P ≥ 0.09). The GAIS analysis resulted in an improvement of 84% (CI: 95% 67-97%) for participants in the Thiamidol group and 74% (CI: 95% 61-93%) for those in the hydroquinone group. There were only mild adverse effects in the Thiamidol group, but allergic contact dermatitis was evidenced in two (8%) participants. CONCLUSION: The melasma improvement achieved using 0.2% Thiamidol did not differ from that of 4% hydroquinone cream. Thiamidol can be considered a suitable option for melasma patients with poor tolerability or treatment failure with hydroquinone.

Drug addiction and alcoholism as predictors for tuberculosis treatment default in Brazil: a prospective cohort study.

This study aimed to evaluate the risk factors for tuberculosis (TB) treatment default in a priority city for disease control in Brazil. A cohort of TB cases diagnosed from 2008 to 2009 was followed up from patients' entry into three outpatient sites, in Juiz de Fora, Minas Gerais (Brazil), until the recording of the outcomes. Drug addiction, alcoholism and treatment site appeared to be independently associated with default. Current users of crack as the hardest drug (odds ratio (OR) 12·25, 95% confidence interval (CI) 3·04-49·26) were more likely to default than other hard drug users (OR 5·67, 95% CI 1·34-24·03), former users (OR 4·12, 95% CI 1·11-15·20) and those not known to use drugs (reference group). Consumers at high risk of alcoholism (OR 2·94, 95% CI 1·08-7·99) and those treated in an outpatient hospital unit (OR 8·22, 95% CI 2·79-24·21%) also were more likely to default. Our results establish that substance abuse was independently associated with default. National TB programmes might be more likely to achieve their control targets if they include interventions aimed at improving adherence and cure rates, by diagnosing and treating substance abuse concurrently with standard TB therapy.

A Self-Powered and Autonomous Fringing Field Capacitive Sensor Integrated into a Micro Sprinkler Spinner to Measure Soil Water Content.

We present here the design and fabrication of a self-powered and autonomous fringing field capacitive sensor to measure soil water content. The sensor is manufactured using a conventional printed circuit board and includes a porous ceramic. To read the sensor, we use a circuit that includes a 10 kHz triangle wave generator, an AC amplifier, a precision rectifier and a microcontroller. In terms of performance, the sensor's capacitance (measured in a laboratory prototype) increases up to 5% when the volumetric water content of the porous ceramic changed from 3% to 36%, resulting in a sensitivity of S = 15.5 pF per unity change. Repeatability tests for capacitance measurement showed that the θ v sensor's root mean square error is 0.13%. The average current consumption of the system (sensor and signal conditioning circuit) is less than 1.5 µ A, which demonstrates its suitability for being powered by energy harvesting systems. We developed a complete irrigation control system that integrates the sensor, an energy harvesting module composed of a microgenerator installed on the top of a micro sprinkler spinner, and a DC/DC converter circuit that charges a 1 F supercapacitor. The energy harvesting module operates only when the micro sprinkler spinner is irrigating the soil, and the supercapacitor is fully charged to 5 V in about 3 h during the first irrigation. After the first irrigation, with the supercap fully charged, the system can operate powered only by the supercapacitor for approximately 23 days, without any energy being harvested.

Meal patterns across ten European countries - results from the European Prospective Investigation into Cancer and Nutrition (EPIC) calibration study.

OBJECTIVE: To characterize meal patterns across ten European countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) calibration study. DESIGN: Cross-sectional study utilizing dietary data collected through a standardized 24 h diet recall during 1995-2000. Eleven predefined intake occasions across a 24 h period were assessed during the interview. In the present descriptive report, meal patterns were analysed in terms of daily number of intake occasions, the proportion reporting each intake occasion and the energy contributions from each intake occasion. SETTING: Twenty-seven centres across ten European countries. SUBJECTS: Women (64 %) and men (36 %) aged 35-74 years (n 36 020). RESULTS: Pronounced differences in meal patterns emerged both across centres within the same country and across different countries, with a trend for fewer intake occasions per day in Mediterranean countries compared with central and northern Europe. Differences were also found for daily energy intake provided by lunch, with 38-43 % for women and 41-45 % for men within Mediterranean countries compared with 16-27 % for women and 20-26 % for men in central and northern European countries. Likewise, a south-north gradient was found for daily energy intake from snacks, with 13-20 % (women) and 10-17 % (men) in Mediterranean countries compared with 24-34 % (women) and 23-35 % (men) in central/northern Europe. CONCLUSIONS: We found distinct differences in meal patterns with marked diversity for intake frequency and lunch and snack consumption between Mediterranean and central/northern European countries. Monitoring of meal patterns across various cultures and populations could provide critical context to the research efforts to characterize relationships between dietary intake and health.

White mold resistance-associated quantitative trait loci in the Jalo x Small White common bean population.

We aimed to identify simple sequence repeat (SSR) markers linked to quantitative trait loci (QTLs) associated with white mold resistance in a segregating population derived from a cross between common bean cultivars Jalo and Small White, in the Southern State of Minas Gerais. Parents were crossed to obtain the F2 generation of 190 plants. From these, F2:3 and F2:4 progenies were obtained for phenotypic evaluation. DNA was extracted from F2 plants and parents for genotyping with SSR primers. For phenotypic assessment by the straw test, we used 190 F2:3 progenies and six lines in a triple-lattice design of 14 x 14 m. Nine trials were conducted for phenotyping by the oxalic acid method to evaluate 177 F2:4 progenies, in addition to the two parents and two common treatments represented by the Jalo and Corujinha lines. The experimental design for the oxalic acid method was completely randomized with three replicates. Adjusted means of evaluations related to F2:3 and F2:4 were used to identify QTLs by using the moving away method from the marker under Bayesian analysis. The markers GATS91, BM197, and X60000 stood out with high heritability and with effects on disease reduction by the straw test; therefore, these markers are promising for selection. The markers BM183, BM189, and SSR-IAC143 were associated with the QTLs identified by oxalic acid, providing greater effects in white mold resistance with high heritability. Based on the oxalic acid and straw test methods, the most prominent marker was PVBR189.

ESPGHAN position paper on management of percutaneous endoscopic gastrostomy in children and adolescents.

OBJECTIVES: This European Society for Pediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) position statement provides a comprehensive guide for health care providers to manage percutaneous endoscopic gastrostomy tubes in a safe, effective, and appropriate way. METHODS: Relevant literature from searches of PubMed, CINAHL, and recent guidelines was reviewed. In the absence of evidence, recommendations reflect the expert opinion of the authors. Final consensus was obtained by multiple e-mail exchange and during 3 face-to-face meetings of the gastroenterology committee of the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition. RESULTS: Endoscopically placed gastrostomy devices are essential in the management of children with feeding and nutritional problems. The article focuses on practical issues such as indications and contraindications. CONCLUSIONS: The decision to place an endoscopic gastrostomy has to be made by an appropriate multidisciplinary team, which then provides active follow-up and care for the child and the device.

Management guidelines of eosinophilic esophagitis in childhood.

OBJECTIVES: Eosinophilic esophagitis (EoE) represents a chronic, immune/antigen-mediated esophageal disease characterized clinically by symptoms related to esophageal dysfunction and histologically by eosinophil-predominant inflammation. With few exceptions, 15 eosinophils per high-power field (peak value) in ≥1 biopsy specimens are considered a minimum threshold for a diagnosis of EoE. The disease is restricted to the esophagus, and other causes of esophageal eosinophilia should be excluded, specifically proton pump inhibitor-responsive esophageal eosinophilia. This position paper aims at providing practical guidelines for the management of children and adolescents with EoE. METHODS: Relevant literature from searches of PubMed, CINAHL, and recent guidelines was reviewed. In the absence of an evidence base, recommendations reflect the expert opinion of the authors. Final consensus was obtained during 3 face-to-face meetings of the Gastroenterology Committee and 1 teleconference. RESULTS: The cornerstone of treatment is an elimination diet (targeted or empiric elimination diet, amino acid-based formula) and/or swallowed, topical corticosteroids. Systemic corticosteroids are reserved for severe symptoms requiring rapid relief or where other treatments have failed. Esophageal dilatation is an option in children with EoE who have esophageal stenosis unresponsive to drug therapy. Maintenance treatment may be required in case of frequent relapse, although an optimal regimen still needs to be determined. CONCLUSIONS: EoE is a chronic, relapsing inflammatory disease with largely unquantified long-term consequences. Investigations and treatment are tailored to the individual and must not create more morbidity for the patient and family than the disease itself. Better maintenance treatment as well as biomarkers for assessing treatment response and predicting long-term complications is urgently needed.

A practical guide for the diagnosis of primary enteric nervous system disorders.

OBJECTIVE: Primary gastrointestinal neuropathies are a heterogeneous group of enteric nervous system (ENS) disorders that continue to cause difficulties in diagnosis and histological interpretation. Recently, an international working group published guidelines for histological techniques and reporting, along with a classification of gastrointestinal neuromuscular pathology. The aim of this article was to review and summarize the key issues for pediatric gastroenterologists on the diagnostic workup of congenital ENS disorders. In addition, we provide further commentary on the continuing controversies in the field. RESULTS: Although the diagnostic criteria for Hirschsprung disease are well established, those for other forms of dysganglionosis remain ill-defined. Appropriate tissue sampling, handling, and expert interpretation are crucial to maximize diagnostic accuracy and reduce interobserver variability. The absence of validated age-related normal values for neuronal density, along with the lack of correlation between clinical and histological findings, result in significant diagnostic uncertainties while diagnosing quantitative aberrations such as hypoganglionosis or ultrashort Hirschsprung disease. Intestinal neuronal dysplasia remains a histological description of unclear significance. CONCLUSIONS: The evaluation of cellular quantitative or qualitative abnormalities of the ENS for clinical diagnosis remains complex. Such analysis should be carried out in laboratories that have the necessary expertise and access to their own validated reference values.

Diagnostic approach and management of cow's-milk protein allergy in infants and children: ESPGHAN GI Committee practical guidelines.

OBJECTIVES: This guideline provides recommendations for the diagnosis and management of suspected cow's-milk protein allergy (CMPA) in Europe. It presents a practical approach with a diagnostic algorithm and is based on recently published evidence-based guidelines on CMPA. DIAGNOSIS: If CMPA is suspected by history and examination, then strict allergen avoidance is initiated. In certain circumstances (eg, a clear history of immediate symptoms, a life-threatening reaction with a positive test for CMP-specific IgE), the diagnosis can be made without a milk challenge. In all other circumstances, a controlled oral food challenge (open or blind) under medical supervision is required to confirm or exclude the diagnosis of CMPA. TREATMENT: In breast-fed infants, the mother should start a strict CMP-free diet. Non-breast-fed infants with confirmed CMPA should receive an extensively hydrolyzed protein-based formula with proven efficacy in appropriate clinical trials; amino acids-based formulae are reserved for certain situations. Soy protein formula, if tolerated, is an option beyond 6 months of age. Nutritional counseling and regular monitoring of growth are mandatory in all age groups requiring CMP exclusion. REEVALUATION: Patients should be reevaluated every 6 to 12 months to assess whether they have developed tolerance to CMP. This is achieved in >75% by 3 years of age and >90% by 6 years of age. Inappropriate or overly long dietary eliminations should be avoided. Such restrictions may impair the quality of life of both child and family, induce improper growth, and incur unnecessary health care costs.

First Report of Tomato chlorosis virus in Potato in Brazil.

Potato plants (Solanum tuberosum cv. Ágata) exhibiting symptoms of leaf roll and interveinal chlorosis, especially on older leaves, were found in a commercial crop in the County of Cristalina, State of Goiás, Brazil in June 2011. The crop was severely infested by whitefly Bemisia tabaci biotype B. Four potato tubers from symptomatic plants were indexed for the presence of the following viruses: Tomato chlorosis virus (ToCV), Potato leaf roll virus (PLRV), Tomato severe rugose virus (ToSRV), and Potato virus Y (PVY). Total RNA was extracted separately from each tuber and used for reverse transcription (RT)-PCR using the HS-11/HS-12 primer pair, which amplifies a fragment of 587 bp from the highly conserved region of the heat shock protein (HSP-70) homolog gene reported for ToCV. The RT-PCR product was subsequently tested by nested-PCR for detection of ToCV with specific primers ToC-5/ToC-6 (2). Amplicons of 463 bp, amplified from total RNA separately extracted from three tubers, were purified and directly sequenced. Comparisons among the three consensus sequences of 448 bp (GenBank Accession Nos. JQ288896, JQ288897, and JQ288898) revealed respectively, 98, 100, and 100% identity with the reported sequence of a tomato isolate of ToCV from Brazil (GenBank Accession No. EU868927) (1). For ToSRV detection, total DNA was extracted from two tubers and a fragment of approximately 820 bp was amplified by PCR with specific primers (3). PLRV and PVY were indexed in two and three tubers, respectively, by double-antibody sandwich-ELISA (SASA, Edinburg, Scotland). Virus-free B. tabaci biotype B were separately transferred to potato and tomato leaves infected with ToCV for an acquisition access period of 24 h. Groups of 30 viruliferous whitefly were transferred to four, young, sprout-grown potato plants cv. Ágata (two plants per virus isolate) for 24-h inoculation access period. After 37 days of inoculation, one plant inoculated with the potato and tomato isolates of ToCV, respectively exhibited symptoms of leaf roll and interveinal chlorosis on order leaves, which were similar to that induced by PLRV. Experimental infection of potato plants with ToCV, which induced leaf roll symptoms resembling PLRV infection, was reported in the United States by Wisler et al. (4). The potato isolate of ToCV was also transmitted by B. tabaci to one of two inoculated tomato plants. The presence of ToCV in all inoculated plants was detected by nested-RT-PCR as described above. To our knowledge, this is the first report on detection of ToCV in field potato plants in the world. Considering that ToCV occurs in innumerous countries around the world, it is transmitted by a cosmopolitan insect, and it induces symptoms similar to PLRV, this finding triggers an alert to field dependent seed-potato multiplication, virus inspector, and certification system. References: (1) J. C. Barbosa et al. Plant Dis. 92:1709, 2008. (2) C. I. Dovas et al. Plant Dis. 86:1345, 2002. (3) F. R. Fernandes et al. Trop. Plant Pathol. 35:43, 2010. (4) G. C. Wisler et al. Plant Dis. 82:270, 1998.

Tomato severe rugose virus: Another Begomovirus Causing Leaf Deformation and Mosaic Symptoms on Potato in Brazil.

Over the past 10 years, Tomato yellow vein streak virus (ToYVSV) has been a major begomovirus in the main solanaceous crop region of Campinas, São Paulo, which includes counties of Sumaré, Monte Mor, Elias Fausto, and Indaiatuba. The top leaves of potato plants (Solanum tuberosum) having deforming mosaic symptoms (dms), which includes a yellow mosaic or mottling on distorted and deformed leaflets, were associated with this geminivirus (4). Recently, a table potato crop (cv. Agata) from Sumaré, with a record of a few or no white flies (Bemisia tabaci), during the winter season of June to September 2006 had 5 to 7% dms, suggestive of seed potato tuber borne virus infection. Double-antibody sandwich (DAS)-ELISA for Potato virus Y (PVY), Potato leaf roll virus (PLRV), Potato virus X (PVX), and Potato virus S (PVS) (SASA kits and protocols, Edinburgh, Scotland) gave negative results for four field collected potato plants showing dms. Bioassays (mechanical transmission from potato leaf extracts in phosphate buffered saline, 1:5 w/v) with test plants of Nicotiana tabaccum cvs. Turkish and TNN, Gomphrena globosa, Chenopodium quinoa, Datura metel, Solanum tuberosum, and a Physalis sp. were all negative. Inoculated D. stramonium developed symptoms resembling ToYVSV infection including vein clearing and mild mottling on new leaves 2 to 3 weeks postinoculation. Using primers PAC1v1978/PAV1c715 for begomovirus detection (3), the predicted PCR amplified fragment of 1,320 bp was obtained from leaf DNA extracted from all four of the dms field potato plants, as well as the inoculated and symptomatic D. stramoniium test plants. Sequence analysis indicated 100% nt identity among the 1.3-kb PCR fragments obtained from potato and D. stramonium infected plants. Sequences of 96 cloned amplicons (pGEM-T Easy Kit; Promega, Madison, WI) from symptomatic plants in the Sumaré potato field were 98 to 99% identical to Tomato severe rugose virus (ToSRV). BLAST analysis of a consensus sequence (Sequencher 3.1; Gene Codes Corporation, Ann Arbor, MI) revealed more than 95 and 99% identity with ToSRV isolates from Uberlandia (Accession No. AY029750) and Goias (Accession No. DQ207749), respectively. The DNA-based phylogenetic dendrogram confirmed the highest similarity with ToSRV and the lowest similarity with ToYVSV (72%), which was located in another cluster. These results indicate that ToSRV was the causal agent producing dms in potato plants from Sumaré. Therefore, similarly to ToYVSV (4), potato dms can be caused by ToSRV. Preliminary tests revealed that ToSRV was transmitted via seed tubers. Thus, it is of concern for seed potato certification in Brazil, especially in the major seed-potato-producing state of Santa Catarina where an outbreak of ToSRV was recently reported in tomato crops (1). Although ToSRV has been identified in other solanaceous crops in Brazil, especially tomato (Lycopersicon esculentum) and sweet pepper (Capsicum annum) (2), to our knowledge, this is the first report of ToSRV in potato in Brazil. Reference: (1) A. T. M. Lima et al. Fitopatol. Bras. 31:224, 2006. (2) D. N. Nozaki et al. Summa Phytopathol. 33:93, 2007. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993. (4) J. A. C. Souza-Dias et al. CultivarHF 5(26):22, 2004.

Simplified dispatch-assisted CPR instructions outperform standard protocol.

OBJECTIVE: Dispatch-assisted chest compressions only CPR (CC-CPR) has gained widespread acceptance, and recent research suggests that increasing the proportion of compression time during CPR may increase survival from out-of-hospital cardiac arrest. We created a simplified CC-CPR protocol to reduce time to start chest compressions and to increase the proportion of time spent delivering chest compressions. This simplified protocol was compared to a published protocol, Medical Priority Dispatch System (MPDS) Version 11.2, recommended by the National Academies of Emergency Dispatch. METHODS: Subjects were randomized to the MPDS v11.2 protocol or a simplified protocol. Data was recorded from a Laerdal Resusci Anne Skillreporter manikin. A simulated emergency medical dispatcher, contacted by cell phone, delivered standardized instructions for both protocols. Outcomes included chest compression rate, depth, hand position, full release, overall proportion of compressions without error, time to start of CPR and total hands-off chest time. Proportions were analyzed by Wilcoxon's Rank Sum tests and time variables with Welch ANOVA and Wilcoxon's Rank Sum test. All tests used a two-sided alpha-level of 0.05. RESULTS: One hundred and seventeen subjects were randomized prospectively, 58 to the standard protocol and 59 to the simplified protocol. The average age of subjects in both groups was 25 years old. For both groups, the compression rate was equivalent (104 simplified versus 94 MPDS, p = 0.13), as was the proportion with total release (1.0 simplified versus 1.0 MPDS, p = 0.09). The proportion to the correct depth was greater in the simplified protocol (0.31 versus 0.03, p < 0.01), as was the proportion of compressions done without error (0.05 versus 0.0, p = 0.16). Time to start of chest compressions and total hands-off chest time were better in the simplified protocol (start time 60.9s versus 78.6s, p < 0.0001; hands-off chest time 69 s versus 95 s, p < 0.0001). The proportion with correct hand position, however, was worse in the simplified protocol (0.35 versus 0.84, p < 0.01). CONCLUSIONS: The simplified protocol was as good as, or better than the MPDS v11.2 protocol in every aspect studied except hand position, and the simplified protocol resulted in significant time savings. The protocol may need modification to ensure correct hand position. Time savings and improved quality of CPR achieved by the new set of instructions could be important in strengthening critical links in the cardiac chain of survival.

Precision analog demodulation technique for open-loop Sagnac fiber optic gyroscopes.

This paper presents an analog technique for the demodulation of interferometric fiber optic gyroscopes signals, based on the calculation of the mean value of the negative cycles of the measured photodetector signal from sinusoidally modulated Sagnac gyroscopes. The developed signal processing circuit was implemented and tested in a laboratory, and it was possible to detect signals as low as 5 microrad. Experimental results measured in a 890 m fiber optic coil gyroscope equipped with the developed circuit showed that the proposed circuit, when compared to the widely used and mature lock-in amplifier technique, presented a 5.1 dB higher signal-to-noise ratio. The circuit is also very stable with the temperature, since a very low drift of 0.04 degrees /h degrees C was measured in a complete gyroscope system. The Brazilian research sounding rocket VSB-30 is currently being equipped with the developed demodulator circuit.

Vascular endothelial growth factor concentrations in the serum and peritoneal fluid of women with endometriosis.

OBJECTIVES: To investigate whether there is an association between vascular endothelial growth factor (VEGF) levels in serum and peritoneal fluid, and the presence of pelvic endometriosis and its clinical symptoms. METHODS: Blood and peritoneal fluid sample levels of VEGF were measured in 46 women undergoing laparoscopy: 32 with suspected endometriosis and 14 with confirmed endometriosis. Data were analyzed according to phase of the menstrual cycle, symptoms, disease stage, and disease site. RESULTS: There were no significant associations between serum and peritoneal fluid levels of VEGF and the presence of endometriosis, even when controlling for the menstrual phase. However, among the women with confirmed endometriosis, there was a significant increase (P=0.002) in the mean peritoneal VEGF level in those in the late secretory phase compared with those in the proliferative and early secretory phases. CONCLUSIONS: Measuring VEGF levels in symptomatic patients is not helpful to differentiate those with endometriosis from those with a different condition. However, in the late secretory and menstrual phases, mean VEGF levels were higher in women with confirmed endometriosis than in those suspected of having the disease.

Effects of the polyamine spermine on binding of follicle-stimulating hormone to membrane-bound immature bovine testis receptors.

The effect of the polyamine, spermine, on the interaction of human 125I-labeled FSH with membrane-bound receptors derived from bovine calf testes has been examined. Concentrations of spermine less than 0.01 M resulted in a slight but insignificant (P greater than 0.10) enhancement of FSH concentrations of 0.01 M and above caused a progressive reduction of FSH binding. Membrane receptors incubated in the presence of spermine at concentrations inhibitory to human 125I-FSH binding (0.01-0.04 M) resulted in an 8-50% decrease in the apparent FSH receptor concentration and a 10-65% decrease in the affinity constant as determined by computerized analysis of the isothermic ligand-binding data. Within the temperature range 4-20 degrees C, simultaneous addition of spermine (0.025 M) increased the reversibility of human 125I-FSH binding approx. 10% (P less than 0.005). Delayed addition of spermine (0.01-0.04 M) resulted in a dose-related dissociation of human 125I-FSH already bound to its receptor (P less than 0.05). However, preincubation of membrane receptors with spermine (0.002-0.04 M) at 4 degrees C or 34 degrees C followed by washing and addition of human 125I-FSH, resulted in an increase in hormone binding (P less than 0.05) over that of controls. If membrane receptor was incubated at 34 degrees C with spermine in the absence of radioligand, the usual loss of hormone binding was reduced (P less than 0.05), while membrane receptor incubated with spermine at 4 degrees C exhibited hormone binding greater (P less than 0.05) than that observed before treatment. Thus, the mechanism of inhibition of human 125I-FSH binding to membrane receptors appears to be correlated with an increase in reversibility of the membrane receptor-human 125I-FSH complex and is expressed as a decrease in the calculated receptor concentration and affinity constant of that interaction. Second, spermine appears to stabilize the membrane receptor in the absence of ligand, presumably through a membrane effect. These data suggest that spermine, and possibly other polyamines, which are endogenous to eukaryotic cells and undergo increases in concentration following stimulation by trophic hormone may play a role in the modulation of the ligand-membrane receptor interaction, in part, through direct effects on the membrane and/or the receptor.

The properties of the secreted gamma-glutamyl hydrolases from H35 hepatoma cells.

gamma-Glutamyl hydrolase has been partially purified and characterized from conditioned culture medium of H35 hepatoma cells. Evidence for heterogeneity of the enzyme is derived from its elution as three distinct peaks of enzymatic activity when the enzyme is purified by TSK-butyl-Sepharose column chromatography. These three enzyme fractions appear to have identical catalytic properties but, as yet, the basis for their resolution is not understood. A rapid, sensitive and simple assay based on reverse-phase HPLC fluorescent detection with pre-column derivatization using o-phthalaldehyde (OPA) was developed to separate OPA-derivatives of poly-gamma-glutamates and glutamic acid. Using this assay and the standard HPLC assay for pteroylpolyglutamates, the enzyme appears to be an endopeptidase with respect to pteroylpenta-gamma-glutamate (PteGlu5), methotrexate penta-gamma-glutamate (4-NH2-10-CH3PteGlu5) and p-aminobenzoyl-penta-gamma-glutamate (pABAGlu5). The initial products are PteGlu1 (or 4-NH2-10-CH3PteGlu1 or pABAGlu1) and intact tetra-gamma-glutamate, which is subsequently degraded to glutamic acid. When penta-gamma-glutamate is the substrate, the cleavage of the gamma-bonds by the enzyme is less ordered, with the early appearance of mono-, di-, tri- and tetraglutamate. Poly-alpha-glutamate is not a substrate nor are pABA-gamma-Glu5 or penta-gamma-glutamate covalently linked to albumin. 4-NH2-10-CH3PteGlu2 or Glu5 bound to dihydrofolate reductase is not a substrate for the enzyme, offering further evidence that protein-associated poly-gamma-glutamates are poor substrates for gamma-glutamyl hydrolase from H35 hepatoma cells.

Progress and approaches in mapping the surfaces of human follicle-stimulating hormone: comparison with the other human pituitary glycoprotein hormones.

The pituitary glycoprotein hormones, including human follicle-stimulating hormone (hFSH), are involved in the physiologic functions of receptor binding, in subunit association during assembly and processing, and in untoward effects such as antibody formation during clinical intervention. These activities derive from specific macromolecular associations; a major research trend has been to map the surfaces of these glycoprotein hormones (hFSH, hLH, hCG, and hTSH) into discrete areas responsible for each activity by using a variety of biochemical approaches. Each surface domain or determinant of the hormone is likely to comprise discontinuous amino acid sequences, from one or both subunits, assembled into a precise, unique, macromolecular surface. The paradigm of antigen-antibody interaction may help to explain how the surfaces are assembled, how the common alpha-subunit combines with the unique beta-subunit of each, and how the receptor interacts with heterodimeric hormone.

Protein synthesis is required for testosterone to decrease ornithine decarboxylase messenger RNA levels in rat Sertoli cells.

We have previously shown that pretreatment of Sertoli cells, derived from 21-day-old Wistar rats, with 5 x 10(-7) M testosterone decreases ornithine decarboxylase (ODC) mRNA levels. In the present study we examined the mechanism of this inhibition. Pretreatment of Sertoli cells for 48 h with 5 x 10(-7) M testosterone decreased ODC mRNA levels 30% at all time points examined. Addition of 25 micrograms/ml cycloheximide (CHX) to the testosterone-pretreated cells resulted in a loss of the testosterone-mediated decrease in ODC mRNA levels by 4 h. Surprisingly, a further 1.8-fold increase in ODC mRNA was observed at 8 h compared to that in untreated cells. Addition of CHX to control cells also resulted in a measurable increase in ODC mRNA levels by 4 h, with a further 1.6-fold increase at 8 h. Identical effects were observed using the protein synthesis inhibitor puromycin (200 micrograms/ml), which has a mechanism of action different from that of CHX. Analysis of the half-life of ODC mRNA after actinomycin-D (5 micrograms/ml) inhibition demonstrated that testosterone did not alter ODC mRNA half-life compared to that in control cells (2.63 vs. 2.55 h). These results demonstrate that the mechanism by which testosterone decreases ODC mRNA levels requires continual protein synthesis, since the effect can be abolished by treating the cells with CHX or puromycin. The data also demonstrate that the testosterone effect is exerted primarily at the transcriptional level, since the half-life of ODC mRNA was not affected by testosterone.