Effects of Salinity Fluctuation Frequency on the Osmolarity,Na^+-K^+-ATPase Activity and HSP70 Expression in Juvenile Chinese Shrimp,Fenneropenaeus chinensis

Experiments were conducted to examine the effects of salinity fluctuation frequency on the osmolarity, Na^＋-K^＋-ATPase activity and HSP70 of Chinese shrimp Fenneropenaeus chinensis ruth initial wet body weight of 1.460g±0.091 g. The salinity in the control group （DO） was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8d, respectively. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8d, respectively, decreased abruptly to salinity 24, lasted for another 2 d, and then was raised to its initial value 28. This was a complete salinity fluctuation cycle that afterwards repeated itself. After 32 days, the osmolarity in treatments D2, D4, D6 and D8 was significantly lower than that in treatment DO （P〈0.05）. There were significant differences in both muscle and eyestalks HSP70 expression among groups. The HSP70 expressions in muscle and eyestalks in group D4 were 61.4% and 57.0% higher, respectively, than that in the control group DO （P〈0.05）. There were, however, no significant differences in gill or hepatopancreas Na^＋-K^＋-ATPase activity between the treatments and the control.

Comparison of the effects of first and second generation silicone hydrogel contact lens wear on tear film osmolarity

AIM:To compare the effects of first and second generation silicone hydrogel(SiH) contact lens wear on tear film osmolarity.METHODS:The healthy subjects who have never used contact lenses before were enrolled in the study.Tear film osmolarity values of 16 eyes(group 1) who wore first generation SiH contact lenses were compared with those of 18 eyes(group 2) who wore second generation SiH contact lenses after three months follow-up.RESULTS:Beforecontactlenswear,tearfilmosmolarity of groups 1 and 2 were 305.02±49.08 milliosmole(mOsm) and 284.66±30.18mOsm,respectively.After three months of contact lens wear,osmolarity values were found 317.74±60.23mOsm in group 1 and 298.40±37.77mOsm in group 2.Although osmolarity values for both groups of SiH contact lens wear after three months periods were slightly higher than before the contact lens wear,the difference was not statistically significant. CONCLUSION:Contact lens wear may cause evaporation from the tear film and can increase tear film osmolarity leading to symptoms of dry eye disease.In the current study,there is a tendency to increase tear film osmolarity for both groups of SiH contact lens wear,but the difference is not statistically significant.

Effects of salinity fluctuation amplitudes on growth,osmolarity,Na^+-K^+-ATPase activity and Hsp70 of juvenile Chinese shrimp Fenneropenaeus chinensis Osbeck

The effects of various salinity fluctuation amplitudes （2, 4, 6 and 8） on the growth, osmolarity, Na＋-K＋-ATPase activity and Hsp70 of juvenile Fenneropenaeus chinensis cultured in seawater with a salinity of 20 were studied. The results show that weight gain in the salinity fluctuation treatments was better than that in control; in particular, the weight gain of treatments S4 and SO, at 231.8% and 196.3%, respectively, was significantly different （P〈0.05）. The hemolymph osmolarity of treatments SO, S2, S4, S6 and S8 was 635.4, 630.8, 623.6, 614.4 and 600.3 mOsm/kg, respectively, and decreased with increasing salinity fluctuation amplitude. The level of Na＋-K＋-ATPase activity in gills ofE chinensis was higher than that in hepatopancreas, but there were no significant differences among all treatments, either in gills or hepatopancreas （P〉0.05）. The relative level of Hsp70 in treatment $4 was 48.4% and 40.4% higher than control in muscle and eyestalks, respectively, with the highest values being recorded under a salinity fluctuation amplitude of 4.

Disrupted ompC Causes Osmosis Sensitivity of Escherichia coli in Alkaline Medium

The Escherichia coli strain DH42 is sensitive to high osmolarity in an alkaline medium. Using mini-Tn5 mutagenesis, construction of mutant strains by homologous recombination and subcloning of DNA fragment techniques, gene ompC was identified as the key factor that, once disrupted, caused osmosis-sensitivity of E. coli strain DH42 grown in an alkaline medium. Through P1 transduction, a mutant strain, D9 （W3110 ompC：kan）, was constructed and growth comparison was performed between DH42 and D9 under different pHs and salt concentrations. The result showed that ompC was necessarily required for hyperosmotic adaptation of E. coli in the alkaline medium.

New approaches for diagnosis of dry eye disease

We reviewed the literature for different diagnostic approaches for dry eye disease(DED) including the most recent advances, contradictions and promising diagnostic tools and technique. We performed a broad literature search for articles discussing different methods for diagnosis of DED including assessment of tear osmolarity, tear film stability, ocular biomarkers and others. Articles indexed in PubMed and google scholar were included. With the growing cosmetic industry, environmental pollution, and booming of digital screens, DED is becoming more prevalent. Its multifactorial etiology renders the diagnosis challenging and invites the emergence of new diagnostic tools and tests. Diagnostic tools can be classified, based on the parameter they measure, into tear film osmolarity, functional visual acuity, tear volume, tear turnover, tear film stability, tear film composition, ocular biomarkers and others. Although numerous methods exist, the most accurate diagnosis can be reached through combining the results of more than one test. Many reported tests have shown potential as diagnostic/screening tools, however, require more research to prove their diagnostic power, alone or in combination. Future research should focus on identifying and measuring parameters that are the most specific to DED diagnosis.

Shellfish allergy and relation to iodinated contrast media: United Kingdom survey

AIM: To assess current practice of United Kingdom cardiologists with respect to patients with reported shellfish/iodine allergy, and in particular the use of iodinated contrast for elective coronary angiography.Moreover we have reviewed the current evidence-base and guidelines available in this area.METHODS: A questionnaire survey was send to 500senior United Kingdom cardiologists(almost 50% cardiologists registered with British Cardiovascular Society)using email and first 100 responses used to analyze practise. We involved cardiologists performing coronary angiograms routinely both at secondary and tertiary centres. Three specific questions relating to allergy were asked:(1) History of shellfish/iodine allergy in pre-angiography assessment;(2) Treatments offeredfor shellfish/iodine allergy individuals; and(3) Any specific treatment protocol for shellfish/iodine allergy cases. We aimed to establish routine practice in United Kingdom for patients undergoing elective coronary angiography. We also performed comprehensive PubMed search for the available evidence of relationship between shellfish/iodine allergy and contrast media.RESULTS: A total of 100 responses were received, representing 20% of all United Kingdom cardiologists. Ninety-three replies were received from consultant cardiologists, 4 from non-consultant grades and 3 from cardiology specialist nurses. Amongst the respondents, 66% routinely asked about a previous history of shellfish/iodine allergy. Fifty-six percent would pre-treat these patients with steroids and anti-histamines. The other 44% do nothing, or do nonspecific testing based on their personal experience as following:(1) Skin test with 1 mL of subcutaneous contrast before intravenous contrast;(2) Test dose 2 mL contrast before coronary injection;(3) Close observation for shellfish allergy patients; and(4) Minimal evidence that the steroid and anti-histamine regime is effective but it makes us feel better.CONCLUSION: There is no evidence that allergy to shellfish alters the risk of reaction to intravenous contrast more than any other allergy and asking about such allergies in pre-angiogram assessment will not provide any additional information except propagating the myth.

Osmotic pressure of serum and cerebrospinal fluid in patients with suspected neurological conditions

Interstitial fluid movement in the brain parenchyma has been suggested to contribute to sustaining the metabolism in brain parenchyma and maintaining the function of neurons and glial cells.The pulsatile hydrostatic pressure gradient may be one of the driving forces of this bulk flow.However,osmotic pressure- related factors have not been studied until now.In this prospective observational study,to elucidate the relationship between osmolality (mOsm/kg) in the serum and that in the cerebrospinal fluid (CSF),we simultaneously measured the serum and CSF osmolality of 179 subjects with suspected neurological conditions.Serum osmolality was 283.6 ± 6.5 mOsm/kg and CSF osmolality was 289.5 ± 6.6 mOsm/kg.Because the specific gravity of serum and CSF is known to be 1.024–1.028 and 1.004–1.007,respectively,the estimated average of osmolarity (mOsm/L) in the serum and CSF covered exactly the same range (i.e.,290.5–291.5 mOsm/L).There was strong correlation between CSF osmolality and serum osmolality,but the difference in osmolality between serum and CSF was not correlated with serum osmolality,serum electrolyte levels,protein levels,or quotient of albumin.In conclusion,CSF osmolarity was suggested to be equal to serum osmolarity.Osmolarity is not one of the driving forces of this bulk flow.Other factors such as hydrostatic pressure gradient should be used to explain the mechanism of bulk flow in the brain parenchyma.This study was approved by the Institutional Review Board of the Tohoku University Hospital (approval No.IRB No.2015-1-257) on July 29,2015.

Survival of rat sciatic nerve segments preserved in storage solutions ex vivo assessed by novel electrophysiological and morphological criteria

Most organ or tissue allografts with viable cells are sto red in solutions ex vivo for hours to seve ral days.Most allografts then require rapid host revascula rization upon transplantation to maintain donor-cell functions(e.g.,cardiac muscle contra ctions,hepatic secretions).In contrast,peripheral nerve allografts stored ex vivo do not require revascularization to act as scaffolds to guide outgrowth by host axons at 1-2 mm/d,likely aided by viable donor Schwann cells.Using current storage solutions and protocols,axons in all these donor orga n/tissue/nerve transplants are expected to rapidly become non-viable due to Wallerian degeneration within days.Therefore,ex vivo storage solutions have not been assessed for preserving normal axonal functions,i.e.,conducting action potentials or maintaining myelin sheaths.We hypothesized that most or all organ storage solutions would maintain axonal viability.We examined several common organ/tissue storage solutions(University of Wisconsin Cold Storage Solution,Normosol-R,Normal Saline,and La ctated Ringe rs) for axonal viability in rat sciatic nerves ex vivo as assessed by maintaining:(1) conduction of artificially-induced compound action potentials;and(2) axonal and myelin morphology in a novel assay method.The ten diffe rent storage solution conditions for peripheral nerves with viable axons(PNVAs) diffe red in their solution composition,osmolarity(250-318 mOsm),temperature(4℃ vs.25℃),and presence of calcium.Compound action potentials and axonal morphology in PNVAs were best maintained for up to 9 days ex vivo in calcium-free hypotonic diluted(250 mOsm) Normosol-R(dNR) at 4℃.Surprisingly,compound action potentials were maintained for only 1-2 days in UW and NS at 4℃,a much shorter duration than PNVAs maintained in 4℃ dNR(9 days) or even in 25℃ dNR(5 days).Viable axons in peripheral nerve allografts are critical for successful polyethylene glycol(PEG)-fusion of viable proximal and distal ends of host axons with viable donor axons to repair segmental-loss peripheral nerve injuries.PEG-fusion repair using PNVAs prevents Wallerian degeneration of many axons within and distal to the graft and results in excellent recovery of sensory/motor functions and voluntary behaviors within weeks.Such PEG-fused PNVAs,unlike all other types of conventional donor transplants,are immune-tolerated without tissue matching or immune suppression.Preserving axonal viability in sto red PNVAs would enable the establishment of PNVA tissue banks to address the current shortage of transplantable nerve grafts and the use of stored PEG-fused PNVAs to repair segmentalloss peripheral nerve injuries.Furthermore,PNVA storage solutions may enable the optimization of ex vivo storage solutions to maintain axons in other types of organ/tissue transplants.

Optimized Conditions for the Delivery of Small Membrane Impermeable Compounds into Human Cells Using Hypotonic Shift

Cell-based assays represent a major end point of high throughput screening (HTS) but a key limitation of such assays is the potentially poor membrane permeability of test compounds. In this study, we optimized the conditions for the delivery of the membrane impermeable compound 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (HPTS) into human cells using hypotonic shift;a method that can promote the uptake of molecules from the extracellular fluid into cell cytoplasm via endocytosis. We showed that uptake of HPTS by cells was a function of hypotonic buffer osmolarity and that delivery was highly efficient with almost 100% of cells displaying uptake. Delivery of HPTS was equally effective at 25°C and 37°C, with delivery of compound proportional to incubation time and concentration of HPTS within the loading medium. The experimental conditions identified in this study could be applied to HTS drug discovery studies providing an effective method of delivering small membrane impermeable compounds into cells.

From Russia with Polyuria

Introduction: Polyuria is a sign for many disease processes, including diabetes mellitus and diabetes insipidus. Urine osmolarity helps distinguish osmotic diuresis caused by diabetes mellitus from water dieresis induced by diabetes insipidus. Case Presentation: We report a case of a 48-year-old woman who presented with polyuria, polydipsia, nocturia, and weight loss after a return from a visit to Russia, during which she received a five-day course of antibiotic Demeclocycline, a tetracycline derivative for dental treatment. She recovered from all clinical manifestations by 8 weeks. Conclusion: Manifestation of transient nephrogenic diabetes insipidus is induced by Demeclocycline.