Family planning (KB) practices and the impact on Papuan women reproductive health.

OBJECTIVE: This study aims to describe and analyze KB or family planning practices and their impact on Papuan women. A case study was conducted in Waena Village, one of the Kampung KB in Jayapura City, Papua. METHODS: The research method used is descriptive qualitative using an ethnographic approach. The determination of the informants was done purposively by assigning one of the key informants. Data collection includes in-depth interviews and observation. Data analysis includes data reduction, data description, and interpretation. RESULTS: The results showed that women of childbearing age carried out the practice of KB by using various forms of birth control. Some use birth control pills and spiral birth control (intrauterine contraception). In addition, the family planning program has provided education to increase knowledge about reproductive health, types of diseases so that women feel more valued and cared for by men with the existence of KB program. Moreover, the existence of the Kampung KB program also has an impact on community social activities, such as educate adolescents and children to have a clean and healthy lifestyle oriented toward family welfare. CONCLUSION: The Kampung KB program in Waena Village has educated the public about reproductive health, which significantly affects community order.

Family planning (KB) practices and the impact on Papuan women reproductive health

Objective: This study aims to describe and analyze KB or family planning practices and their impact on Papuan women. A case study was conducted in Waena Village, one of the Kampung KB in Jayapura City, Papua. Methods: The research method used is descriptive qualitative using an ethnographic approach. The determination of the informants was done purposively by assigning one of the key informants. Data collection includes in-depth interviews and observation. Data analysis includes data reduction, data description, and interpretation. Results: The results showed that women of childbearing age carried out the practice of KB by using various forms of birth control. Some use birth control pills and spiral birth control (intrauterine contraception). In addition, the family planning program has provided education to increase knowledge about reproductive health, types of diseases so that women feel more valued and cared for by men with the existence of KB program. Moreover, the existence of the Kampung KB program also has an impact on community social activities, such as educate adolescents and children to have a clean and healthy lifestyle oriented toward family welfare. Conclusion: The Kampung KB program in Waena Village has educated the public about reproductive health, which significantly affects community order. (AU)

Air assisted emulsification liquid-liquid microextraction based on deep eutectic solvent for preconcentration of methadone in water and biological samples.

Nowadays, a deep eutectic solvent (DES) has recently been considered as a green ion liquid analogue. In this study, a new compound of DES has been synthesized as an extraction solvent in air assistedemulsification liquid-liquid microextraction method (DES-AAELLME) for preconcentration and extraction of methadone followed by gas chromatographyflame ionization detector (GC-FID). To obtain an efficient water-miscible deep eutectic solvent, choline chloride (ch-cl) and 5,6,7,8-Tetrahydro-5,5,8,8-tetramethylnaphthalen-2-ol (TNO) were mixed at a molar ratio of 1:2 and tetrahydrofuran (THF) was used as a demulsifier solvent into homogeneous solution for providing a turbid state. The solution was rapidly sucked up and injected 10 times using a 10-mL glass syringe to enhance the turbidity of solution and disperse the aggregated DES droplets into aqueous phase. Some important parameters affecting extraction recovery were investigated. Under optimum conditions, the calibration curve was linear in the concentration range from 2 to 8000µgL-1. The limit of detection and the limit of quantification were found 0.7µgL-1 and 2.3µgL-1 respectively with preconcentration factor of 270. The precision, as the relative standard deviation (RSD) (n=6), was <6% respectively. This method was successfully applied to determine methadone in water and biological samples with an appropriate recovery about 98.4-101.2%.

Development of air assisted liquid phase microextraction based on switchable-hydrophilicity solvent for the determination of palladium in environmental samples.

A novel switchable-hydrophilicity solvent (SHS) in combination with air-assisted liquid-liquid microextraction (AALLME) which called AA-SHS-LPME, has been applied for preconcentration of Pd(II). A water-immiscible solvent, Triethylamine (TEA), was used as a green switchable solvent that switched reversibly between one form that was miscible with water to another that formed a biphasic mixture with water. CO2 was used as a stimulus for triggering the transformation to the water-miscible form and removal of CO2 to achieve the reverse. Separation was carried out by addition of sodium hydroxide, which produced a change on the ionization state of amine. AALLME led to the rapid formation of fine droplets of the extractant in the aqueous solution, and the contact surface between both immiscible liquids was significantly enlarged. Detection limit value, preconcentration factor and relative standard deviation (RSD, n=10) were found 0.07µgL(-1), 64 and 3.5% respectively. The recovery of the analyte in water, road dust and catalytic converter samples was in the range of 98.3-103.2%. The obtained results indicated that the developed method could be an efficient analytical method for the routine analysis in the environmental field.

Magnetic Graphene-Dispersive Solid-Phase Extraction for Preconcentration and Determination of Lead and Cadmium in Dairy Products and Water Samples.

A magnetic-dispersive solid-phase extraction (MDSPE) was used for precocentration of Pb(II) and Cd(II) in milk, yoghurt and water samples. An appropriate amount of suspension containing the magnetic graphene and Triton X-114 was injected rapidly into the aqueous sample by a syringe. Triton X-114 was used to achieve stable suspension of graphene in solution. The structure of the resulting products was confirmed by Fourier transform infrared (FT-IR) spectra, X-ray diffraction (XRD) spectrometry. The effects of various parameters were studied. A detection limit of 0.16 and 0.50 µg L(-1) for Cd(II) and Pb(II) was obtained, respectively. The relative standard deviations (RSDs, n = 10) of 50 µg L(-1) of Pb and Cd were 3.3 % and 2.1 %, respectively. The results indicated that the present method can be reliably used for determination of Pb(II) and Cd(II) in dairy products and water samples with good recoveries.

Interaction of 5-Fluorouracil and its derivatives with bovine serum albumin.

Fluorouracil (5-FU) and its derivatives are the most commonly used drugs to treat many types of cancer. Two dual functional agents, FUPAE and FUPAP, derived from 5-Fluorouracil (5-FU) have shown radiosensitizing activity but unlike their components were not cytotoxic. This study was designed to examine the interaction of BSA with 5-Fluorouracil (5-FU) and two of its derivatives; FUPAE and FUPAP at physiological conditions, using a constant protein concentration and various drug contents. FTIR, UV-Vis spectroscopic methods as well as molecular modelling were used to determine the drugs binding mode, the binding constants and the effects of drug complexation on BSA stability and conformation. Structural analysis showed that 5-Fluorouracil, FUPAE and FUPAP bind BSA via polypeptide polar groups with overall binding constants of K(5-FU-BSA)=3.02(±0.09)×10(3), K(FUPAE-BSA)=1.08(±0.04)×10(4), K(FUPAP-BSA)=1.21(±0.06)×10(4) M(-1). BSA conformation was altered by a major reduction of α-helix from 69% (free BSA) to 34% with 5-FU, 40% with FUPAE, 38% with FUPAP. These results suggest that serum albumins might act as carrier proteins for FUPAE and FUPAP in delivering them to target tissues.

Determination of some phenolic compounds in Crocus sativus L. corms and its antioxidant activities study.

It is well known that phenolic compounds are constituents of many plants. In this study, the total phenolics content in Crocus sativus L. corms in dormancy and waking stages were determined by the Folin-Ciocalteu method. Analysis was carried out by gas chromatography-mass spectrometry (GC-MS) after silylation by N-methyl-N-trimethylsilyl trifluroacetamide (MSTFA) + %1 trimethyl iodosilane (TMIS). Numerous compounds were detected and 11 compounds were identified. The highest phenolics content in waking corms was observed for gentisic acid (5.693 ± 0.057 µg/g) and the lowest for gallic acid (0.416 ± 0.006 µg/g); also these two phenolic compounds are the highest (0.929 ± 0.015 µg/g) and lowest (0.017 ± 0.001 µg/g) phenolics in dormant corms, respectively. The results from quantization and GC-MS analysis showed a high concentration of phenolic compounds in waking corms than the dormant stage. Furthermore, the radical scavenging activities of saffron corms were studied by 1,1-diphenyl-2-pycrylhydrazyl (DPPH) test and EC (50)values were determined about 2055 ppm and 8274 ppm for waking and dormant corms, respectively.

Biological treatment of toluene contaminated wastewater by Alcaligenese faecalis in an extractive membrane bioreactor; experiments and modeling.

Conventional wastewater treatment methods are not efficient in treating wastewaters contaminated with volatile hydrocarbons such as benzene, toluene and xylenes (BTX). The aim of this study is to enhance the efficiency of an extractive membrane bioreactor (EMBR) in treating toluene contaminated wastewater by usage of pure culture of Alcaligenese faecalis. Toluene was used as a model of toxic contaminant because of its wide presence in wastewaters contaminated with petrol derivatives. The Haldane kinetic model adequately described the dynamic behavior of the toluene biodegradation by the strain of A. faecalis over a wide range of initial toluene concentrations (50-1,000 mg L(-1)) with kinetic constants micro(max) = 0.066 h(-1), k(s) = 91.7 mg/L and k(I) = 278.2. Overall mass transfer coefficient has been measured and described as resistance in the series model. No biofilm formed on the exterior surface of the membrane; however in previous works the layer of the biofilm on the exterior surface of the membrane acts as a mass transfer resistance. A mathematical model was developed to predict the pollutant concentration profile along the tube side of the membrane modules.

Antigen-activated T cells induce IL-12p75 production from dendritic cells in an IFN-γ-independent manner.

The addition of IL-12p75 to naïve CD4(+) T cells promotes their differentiation towards a TH1-type cytokine pattern. Dendritic cells stimulated by LPS generate IL-12p75, but only if the environment also contains IFN-γ. Thus, it appears that IFN-γ is needed to start the response that will result in further production of IFN-γ. We previously reported that paradoxically DCs produce IL-12p75 only after engaging primed, but not naïve T cells. This study examines the mechanism by which primed T cells trigger IL-12p75 secretion and asks whether this induction is also dependent on the presence of IFN-γ. Here, we show that, in contrast to LPS, primed T cells induce IL-12p75 in an IFN-γ-independent manner. Addition of rIFN-γ to cocultures of naïve T cells with DCs did not induce IL-12p75. Moreover, antigen-activated CD4(+) T cells from wild type or IFN-γ-deficient mice both initiated IL-12p75 production from DCs. Surprisingly, we found that synergies between three T-cell-derived factors - CD40 Ligand, IL-4 and GM-CSF - were necessary and sufficient for IL-12p75 production. These results suggest that there are at least two distinct pathways for IL-12p75 production in vivo. Furthermore, the T-cell-dependent pathway of IL-12p75 production employs molecules that are not classically associated with a TH1-type response.

Decreased polyunsaturated and increased saturated fatty acid concentration in spermatozoa from asthenozoospermic males as compared with normozoospermic males.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied fatty acid composition of the phospholipids in spermatozoa in asthenozoospermic and normozoospermic men and determined the ratio of polyunsaturated fatty acids (PUFAs) to saturated fatty acids of spermatozoa of these two groups. Fatty acid concentration of spermatozoa was determined in 15 asthenozoospermic and eight normozoospermic semen samples by thin layer chromatography and gas chromatography. The most abundant polyunsaturated and saturated fatty acids in normozoospermic samples were docosahexaenoic acid (DHA 22 : 6 omega3, 98.5 +/- 4.5 nmol per 10(8) spermatozoa, mean +/- SE) and palmitic acid (103 +/- 17 nmol per 10(8) spermatozoa) respectively. The mean +/- SE values of DHA and palmitic acid in asthenozoospermic samples were 53.9 +/- 11.6 and 145 +/- 14.7 nmol per 10(8) spermatozoa respectively. Compared with normozoospermic samples, asthenozoospermic samples showed lower levels of PUFA and higher amount of saturated fatty acids. The mean +/- SE ratios of sperm PUFA/saturated fatty acids in asthenozoospermic and normozoospermic samples were 0.66 +/- 0.06 and 1.45 +/- 0.16 (P < 0.001) respectively. This study demonstrates that spermatozoa of asthenozoospermic men have lower levels of PUFA compared with saturated fatty acids. This may be contributory to the poor motility noted in samples from these men.

T-cell control of IL-12p75 production.

It is currently thought that IL-12, produced by dendritic cells (DC) early after stimulation by bacterial pathogens or lipopolysaccharide (LPS), acts as a pro-inflammatory cytokine bridging the innate and adaptive immune responses. We found, however, that it is only the p40 subunit and not the IL-12p75 heterodimer that is secreted early in copious amounts in response to LPS. Neither naïve T cells, nor a variety of microbial products, were able to induce IL-12p75 production unless the DC were conditioned by the presence of interferon-gamma (IFN-gamma) or by encounter with previously activated T cells. The inability of naïve T cells or of bacterial products to induce IL-12 argues against its early role as the initiator of innate and adaptive immune responses.

IL-12: the role of p40 versus p75.

Interleukin (IL)-12p75 is a heterodimeric cytokine composed of the product of two different genes that specify p35 and p40 subunits. The prevailing view is that IL-12 acts as a proinflammatory cytokine that bridges the innate and adaptive immune responses and skews T-cell reactivity toward a TH1 cytokine pattern. Though the terms IL-12 and IL-12p40 are often used interchangeably, and measurements of the p40 chain are often interpreted as measurements of the intact p75 heterodimer, such interchangeable usage may be incorrect. In the following discussion, I will delineate an alternative hypothesis for the roles of the p40 and p75 proteins, suggesting specifically, that: (1) in vivo, secretion of free p40 precedes that of p75 in response to pathogens; (2) induction of p40 is a T-independent response by antigen presenting cells (APCs) to early host-pathogen interactions; and (3) IL-12p75 is a late product, whose induction requires T-dependent signals. It is made as a result, rather than as a cause, of TH1 differentiation. Thus, it is the p40 protein, either alone or paired with other polypeptides, rather than p75, that acts as an interface between the innate and adaptive immune responses.

CTL generation in the presence of IL-4 is inhibited by free p40: evidence for early and late IL-12 function.

The cytokine requirements for the generation of a CTL response were examined using thymocyte responders stimulated with Con A. In agreement with previous reports, IL-4 alone can generate a strong CTL response. To determine whether IL-12 was required for this response, we added either a neutralizing Ab against IL-12 or an antagonist of IL-12 function, p40. Both anti-IL-12 and p40 were found to inhibit the generation of CTL in the presence of IL-4. Consistent with these data, exogenous IL-12 was found to synergize with IL-4, particularly, when IL-4 was added at levels unable to generate a CTL response alone. This synergistic response was observed whether IL-12 was added at the beginning of culture along with IL-4 or after the first day to cultures initiated with IL-4. Likewise, the combination of IL-2 plus IL-12 evoked a strong synergistic response when cultures were initiated with IL-2 and IL-12 was added initially or after the first day of culture. Addition of either IL-2 or IL-12 alone did not generate a response. In contrast to these data showing that IL-12 was able to signal late in culture, p40 inhibited CTL generation in response to IL-4 only when added at the beginning of culture. These results imply that CTL induction involves a crucial early IL-12 signal, which once delivered permits cells to respond to a later IL-12 signal. These data are consistent with the idea that free circulating p40 could inhibit the generation of CTL by antagonizing the early IL-12 "differentiation" signal.

LFA-1 and L-selectin regulation of recirculating lymphocyte tethering and rolling on lung microvascular endothelium.

Recirculating lymphocytes migrate into areas of lung inflammation by binding to microvascular endothelium and transmigrating into extravascular tissue. In this report, we examined the multiple-step paradigm using a unique system: recirculating lymphocytes from sheep peripheral lymphatics adhering to activated lung microvascular endothelium in conditions of physiologic flow. Video microscopy demonstrated that recirculating lymphocytes formed abrupt adhesions, without requisite rolling, on the lung microvascular endothelial cells. Lymphocyte velocity was unchanged within 100 ms of the development of firm adhesions. To dissect the adhesion mechanism, the lymphocytes were pretreated with anti-LFA-1 or anti-L-selectin monoclonal antibody (mAb). Both mAb decreased the incidence of firm adhesions. The mechanism of this inhibition was investigated using time-lapse topographic reconstructions of cell movement after pretreatment with mAb. Time-lapse analysis of the movement of lymphocytes pretreated with anti-LFA-1 mAb suggested that abortive adhesion was manifested by a characteristic saltatory movement and a sustained reduction in cell velocity (rolling) to <25 microns/s. In contrast, abortive adhesions of lymphocytes pretreated with anti-L-selectin mAb demonstrated transient arrest (tethering) but minimal rolling before resumption of baseline velocity in the flow stream. These observations provide insights into selectin and integrin regulation of lymphocyte transmigration into the lung. Further, the results of mAb inhibition suggest that the mechanism of lymphocyte migration may have some unique features not observed in studies of neutrophil transmigration.

Expression of membrane glycoconjugates on sheep lung endothelium.

BACKGROUND: Carbohydrates play an important role in both the regulation and expression of endothelial cell surface molecules. EXPERIMENTAL DESIGN: To investigate the molecular phenotype of endothelial cells in the sheep lung, we used a panel of 39 lectins and carefully titrated lectin histochemistry to identify small vessel glycoconjugates. Expression of glycoconjugates on isolated cells was studied by flow cytometry and lectin Western blotting. RESULTS: Eight lectins stained small vessel endothelium with minimal background staining. To confirm cell surface binding, endothelial cells from the peripheral lung were isolated and tested for lectin recognition by flow cytometry. Three lectins (Datura stramonium, Griffonia simplicifolia-1, Lycopersicon esculentum) also stained isolated lung cells by flow cytometry. A lectin Western transfer technique demonstrated common binding to a high molecular weight (100 to 130 kD) band. CONCLUSIONS: The use of lectins as probes of cell surface carbohydrate expression supports the possibility of selective glycoconjugate expression on sheep endothelium.

In situ fluorescence labeling of sheep lung microvascular endothelium.

Endothelial cells are intimately involved in a variety of biological processes such as inflammatory disorders, wound healing, and tumor invasion. The finding of endothelial heterogeneity in various tissues has led to major efforts to isolate and culture microvascular endothelial cells in human and animal tissue. In this report we have used phosphatidyl ethanolamine (PE)-labeled liposomes to fluorescently label the sheep lung microvasculature in situ. Using normotensive perfusion pressure, the PE-labeled liposomes did not extravasate into extravascular lung tissue. Mechanical and enzymatic digestion of the lung tissue demonstrated that the PE-labeled liposomes provided a stable label of the vascular lining cells during ex vivo processing. After digestion, the overwhelming majority of the fluorescent label appeared in cellular aggregates. Approximately 80% of these cells demonstrated an in vitro phenotype consistent with microvascular endothelium. A novel monoclonal antibody selective for sheep endothelial cells was developed to confirm the presence of lung endothelium in the fluorescently labeled cellular aggregates. We conclude that in situ fluorescence labeling of vascular lining cells provides an anatomic marker for relevant vascular lining cells and an opportunity to study these cells in vitro.

Hybridoma screening using an amplified fluorescence microassay to quantify immunoglobulin concentration.

The early identification of antibody-secreting hybridomas significantly reduces the time and resources devoted to unproductive colonies. To quantify the immunoglobulin concentration in hybridoma supernatants within 2 weeks of fusion, we used immunomagnetic microspheres to capture immunoglobulin in the hybridoma culture supernatant. The captured immunoglobulin was detected using a goat anti-mouse second antibody liked to beta-galactosidase. With data transformation to correct for the nonlinear accumulation of the fluorescent reaction product, the enzymatic hydrolysis of fluorescein digalactoside permitted the reliable detection of less than 10 pg of immunoglobulin per milliliter. To determine the value of quantifying immunoglobulin concentration within 2 weeks of fusion, the amplified fluorescence microassay was applied to the evaluation of 3 consecutive fusions and more than 1200 growing hybridoma colonies. Using antibody concentrations greater than 10 ng/ml as a threshold for routine subculture, the selection of hybridoma colonies based on antibody secretion was threefold more efficient than selection based on colony growth alone. These results suggest the utility of the early determination of immunoglobulin concentration in the selection of hybridoma colonies.

Affinity membrane identification of immunoglobulin subclass in hybridoma screening.

Monoclonal hybridomas secrete immunoglobulins with a single antigen specificity and distinct class/subclass structure. Hybridoma management has commonly incorporated tests of antigen specificity into early screening procedures, but has not typically utilized assays of immunoglobulin structure. In this article, we describe a technique of class/subclass typing using polyvinylidene difluoride affinity membranes and a colorigenic enzymatic amplification system. The typing of monoclonal antibody structure was sufficiently sensitive to permit its routine use within several weeks of hybridoma fusion. The information obtained from early and routine class/subclass determinations included a semiquantitative assessment of monoclonal antibody concentration. In addition, the detection of a single immunoglobulin class/subclass in a microtiter well supernatant supported the possibility that the colony was monotypic. The application of class/subclass typing and Poisson statistics to hybridoma fusions provided a numerical estimate of the probability of colony monotypia.

Membrane glycoconjugates expressed on sheep airway epithelium.

In mammalian lung, selective airway inflammatory reactions have been associated with viral infections, transplant rejection, and autoimmune diseases. Although the molecular basis for this selective reactivity is unknown, the importance of carbohydrates in immunologic processes suggests a potential role for membrane glycoconjugates in tissue-specific inflammatory reactions. In the present work we examined a panel of 39 lectins for their pattern of reactivity in the peripheral airways of the sheep lung. The size of the panel facilitated a comprehensive description of the glycoconjugate localization on the airway epithelium. Four lectins (agglutinins for Helix aspersa, Psophocarpus tetragonolobus, Trichosanthes kirilowii, and Griffonia simplifolia II) revealed selective reactivity with the small airway epithelium. On lectin Western blotting, these four lectins demonstrated a common low molecular weight banding profile that was distinct from control lectins. The histochemical staining patterns and Western blotting profiles provided evidence for the selective expression of membrane glycoconjugates in the peripheral airways of the sheep lung.