Intraocular pressure following 18 hours of systemic dehydration in ocular normotensive healthy subjects

Aim: To investigate the effect of dehydration on intraocular pressure (IOP). Subjects: The study population comprised 50 subjects (27 men and 23 women) with a mean age of 42.84 ± 2.64 years. Method: The subjects dehydrated for 18 hours. IOP was assessed in both eyes using the Perkins handheld applanation tonometer. Baseline measurement of IOP was taken before the subjects dehydrated for 18 hours and repeated after dehydration. Blood samples were taken before and after dehydration for the determination of plasma osmolality and plasma antidiuretic hormone (ADH) concentrations. Results: A statistically significant (p < 0.001) reduction in the mean weight of subjects occurred after dehydration. There was a significant reduction in IOP from 16.68 mmHg ± 0.32 mmHg to 13.08 mmHg ± 0.31 mmHg after dehydration (p < 0.001). Mean plasma osmolality showed a statistically significant increase (p < 0.05) from baseline of 290.02 mOsmol/kg ± 1.25 mOsmol/kg to 294.96 mOsmol/kg ± 1.33 mOsmol/kg after dehydration. There was also a significant increase (p < 0.05) in mean plasma ADH concentration from 5.36 pg/mL ± 0.21 pg/mL to 6.40 pg/mL ± 0.20 pg/mL after dehydration. There were no significant differences in both systolic and diastolic blood pressures before and after dehydration. Conclusion: Dehydration decreases IOP in healthy humans; this decrease probably results from an increase in plasma osmolality and plasma ADH concentrations

Thirst Perception and Drinking in Euhydrate and Dehydrated Humans

Summary: Studies on how the body senses the need to correct extracellular and intracellular volumes and ionic concentration changes is relatively scanty. The present studies were designed to determine the effect of oral distilled water (DW) and saline loads, gargling with DW and DW preload on thirst perception (TP) and drinking in euhydrate and dehydrated subjects. The subjects were healthy male volunteers between the ages of 17 and 35 years. Group A subjects were given DW or various concentrations of sodium chloride (NaCl) orally. Subjects in groups B, C and D were dehydrated for 18 hours before the experiment. Group B gargled 500ml of DW in divided volume of 50ml at five minutes interval over a period of 50 minutes. Group C gargled with DW and different concentrations of NaCl. Group D were preloaded with four volumes of DW before ad libitum DW intake. TP was rated using the Visual Analogue Scale. Results showed that in Group A, drinking DW reduced TP, suggesting that baseline TP in normal euhydrate subjects is slightly elevated. Drinking DW reduced TP more than drinking NaCl solutions. Gargling resulted in a gradual fall in TP. The decrease in TP was statistically significant after 30 minutes of gargling. Gargling with different concentrations of NaCl solutions resulted in significant reductions in TP in all the groups. There was a significant decrease in TP in the group preloaded with 1000ml of distilled water at 5 minutes of rehydration. At 20 minutes TP was abolished suggesting that approximately 1000ml of water was needed for the rehydration. These results show that baseline TP in euhydrates is elevated and that TP increases in dehydrated subjects. Gargling reduces TP, but did not abolish thirst. It is suggested that a fall in plasma osmolality due to drinking may be responsible for abolishing thirst

Polymerase-directed synthesis of C5-ethynyl locked nucleic acids

Modified nucleic acids have considerable potential in nanobiotechnology for the development of nanomedicines and new materials. Locked nucleic acid (LNA) is one of the most prominent nucleic acid analogues reported so far and we herein for the first time report the enzymatic incorporation of LNA-U and C5-ethynyl LNA-U nucleotides into oligonucleotides. Phusion High Fidelity and KOD DNA polymerases efficiently incorporated LNA-U and C5-ethynyl LNA-U nucleotides into a DNA strand and T7 RNA polymerase successfully accepted the LNA-U nucleoside 5′-triphosphate as substrate for RNA transcripts

Increasing the Analytical Sensitivity by Oligonucleotides Modified with Para- and Ortho-Twisted Intercalating Nucleic Acids – TINA

The sensitivity and specificity of clinical diagnostic assays using DNA hybridization techniques are limited by the dissociation of double-stranded DNA (dsDNA) antiparallel duplex helices. This situation can be improved by addition of DNA stabilizing molecules such as nucleic acid intercalators. Here, we report the synthesis of a novel ortho-Twisted Intercalating Nucleic Acid (TINA) amidite utilizing the phosphoramidite approach, and examine the stabilizing effect of ortho- and para-TINA molecules in antiparallel DNA duplex formation. In a thermal stability assay, ortho- and para-TINA molecules increased the melting point (Tm) of Watson-Crick based antiparallel DNA duplexes. The increase in Tm was greatest when the intercalators were placed at the 5′ and 3′ termini (preferable) or, if placed internally, for each half or whole helix turn. Terminally positioned TINA molecules improved analytical sensitivity in a DNA hybridization capture assay targeting the Escherichia coli rrs gene. The corresponding sequence from the Pseudomonas aeruginosa rrs gene was used as cross-reactivity control. At 150 mM ionic strength, analytical sensitivity was improved 27-fold by addition of ortho-TINA molecules and 7-fold by addition of para-TINA molecules (versus the unmodified DNA oligonucleotide), with a 4-fold increase retained at 1 M ionic strength. Both intercalators sustained the discrimination of mismatches in the dsDNA (indicated by ΔTm), unless placed directly adjacent to the mismatch – in which case they partly concealed ΔTm (most pronounced for para-TINA molecules). We anticipate that the presented rules for placement of TINA molecules will be broadly applicable in hybridization capture assays and target amplification systems

FRA-1 protein overexpression is a feature of hyperplastic and neoplastic breast disorders

BACKGROUND: Fos-related antigen 1 (FRA-1) is an immediate early gene encoding a member of AP-1 family of transcription factors involved in cell proliferation, differentiation, apoptosis, and other biological processes. fra-1 gene overexpression has an important role in the process of cellular transformation, and our previous studies suggest FRA-1 protein detection as a useful tool for the diagnosis of thyroid neoplasias. Here we investigate the expression of the FRA-1 protein in benign and malignant breast tissues by immunohistochemistry, Western blot, RT-PCR and qPCR analysis, to evaluate its possible help in the diagnosis and prognosis of breast neoplastic diseases. METHODS: We investigate the expression of the FRA-1 protein in 70 breast carcinomas and 30 benign breast diseases by immunohistochemistry, Western blot, RT-PCR and qPCR analysis. RESULTS: FRA-1 protein was present in all of the carcinoma samples with an intense staining in the nucleus. Positive staining was also found in most of fibroadenomas, but in this case the staining was present both in the nucleus and cytoplasm, and the number of positive cells was lower than in carcinomas. Similar results were obtained from the analysis of breast hyperplasias, with no differences in FRA-1 expression level between typical and atypical breast lesions; however the FRA-1 protein localization is mainly nuclear in the atypical hyperplasias. In situ breast carcinomas showed a pattern of FRA-1 protein expression very similar to that observed in atypical hyperplasias. Conversely, no FRA-1 protein was detectable in 6 normal breast tissue samples used as controls. RT-PCR and qPCR analysis confirmed these results. Similar results were obtained analysing FRA-1 expression in fine needle aspiration biopsy (FNAB) samples. CONCLUSION: The data shown here suggest that FRA-1 expression, including its intracellular localization, may be considered a useful marker for hyperplastic and neoplastic proliferative breast disorders

Effect of chemical modifications on modulation of gene expression by duplex antigene RNAs that are complementary to non-coding transcripts at gene promoters

Antigene RNAs (agRNAs) are small RNA duplexes that target non-coding transcripts rather than mRNA and specifically suppress or activate gene expression in a sequence-dependent manner. For many applications in vivo, it is likely that agRNAs will require chemical modification. We have synthesized agRNAs that contain different classes of chemical modification and have tested their ability to modulate expression of the human progesterone receptor gene. We find that both silencing and activating agRNAs can retain activity after modification. Both guide and passenger strands can be modified and functional agRNAs can contain 2′F-RNA, 2′OMe-RNA, and locked nucleic acid substitutions, or combinations of multiple modifications. The mechanism of agRNA activity appears to be maintained after chemical modification: both native and modified agRNAs modulate recruitment of RNA polymerase II, have the same effect on promoter-derived antisense transcripts, and must be double-stranded. These data demonstrate that agRNA activity is compatible with a wide range of chemical modifications and may facilitate in vivo applications

Blood pressure patterns in rural, semi-urban and urban children in the Ashanti region of Ghana, West Africa

BACKGROUND: High blood pressure, once rare, is rapidly becoming a major public health burden in sub-Saharan/Africa. It is unclear whether this is reflected in children. The main purpose of this study was to assess blood pressure patterns among rural, semi-urban, and urban children and to determine the association of blood pressure with locality and body mass index (BMI) in this sub-Saharan Africa setting. METHODS: We conducted a cross-sectional survey among school children aged 8–16 years in the Ashanti region of Ghana (West-Africa). There were 1277 children in the study (616 boys and 661 females). Of these 214 were from rural, 296 from semi-urban and 767 from urban settings. RESULTS: Blood pressure increased with increasing age in rural, semi-urban and urban areas, and in both boys and girls. The rural boys had a lower systolic and diastolic blood pressure than semi-urban boys (104.7/62.3 vs. 109.2/66.5; p < 0.001) and lower systolic blood pressure than urban boys (104.7 vs. 107.6; p < 0.01). Girls had a higher blood pressure than boys (109.1/66.7 vs. 107.5/63.8; p < 0.01). With the exception of a lower diastolic blood pressure amongst rural girls, no differences were found between rural girls (107.4/64.4) and semi-urban girls (108.0/66.1) and urban girls (109.8/67.5). In multiple linear regression analysis, locality and BMI were independently associated with blood pressure in both boys and girls. CONCLUSION: These findings underscore the urgent need for public health measures to prevent increasing blood pressure and its sequelae from becoming another public health burden. More work on blood pressure in children in sub-Saharan African and other developing countries is needed to prevent high blood pressure from becoming a major burden in many of these countries

Variations in the prevalence of point (pre)hypertension in a Nigerian school-going adolescent population living in a semi-urban and an urban area

<p>Abstract</p> <p>Background</p> <p>Hypertension has been shown to start in early life and to track into adulthood. Detecting adolescents with hypertension and prehypertension will aid early intervention and reduce morbidity and mortality from the disorders. This study reports the point-prevalence of the two disorders in a semi-urban and an urban population of school-going adolescents in Nigeria.</p> <p>Methods</p> <p>A total of 843 adolescents from two places of domicile were studied. Their blood pressures and anthropometric indices were measured using standard protocol. Point-hypertension and point-prehypertension were defined with respect to each subject's gender, age and height. The prevalence of the disorders was calculated and reported age-wise and nutritional status-wise.</p> <p>Results</p> <p>The prevalence of point-prehypertension in the semi-urban area was 22.2% (20.7% for girls and 23.1% for boys) while it was 25.0% (21.8% for girls and 29.2% for boys) in the urban area. The prevalence of point-hypertension was 4.6% (4.1% for girls and 4.8% for boys) in the semi-urban area and 17.5% (18.0% for girls and 16.9% for boys) in the urban area. Point-prehypertension was not detected among the thin subjects of both places of domicile. The prevalence of point-prehypertension was similar in both the urban and semi-urban areas among the subjects who had normal BMI-for-age, and over-weight/obese subjects respectively. From the semi-urban to the urban area, the prevalence of point-hypertension increased approximately 3-folds among thin and normal BMI-for-age subjects, and 10-folds among overweight/obese subjects. Systolic hypertension was more preponderant in both the semi-urban and urban areas.</p> <p>Conclusions</p> <p>The prevalence of both disorders is considerably high in the studied populations. Urgent pediatric public health action is needed to address the situation.</p