Razlika između proteina sjemene plazme i proteina sperme za dobru i lošu sposobnost smrzavanja ejakulata nerasta

The present study was performed to compare the expression of sperm proteins, i.e. triosephosphate isomerase (TPI) and acrosin binding protein (ACRBP) and seminal plasma proteins, i.e. glutathione peroxidase 5 (GPX5) and fibronectin 1 (FN1), in boar semen with good, moderate and poor freezability. The study was conducted by determining the protein contents in 32 sperm samples and 38 seminal plasma samples of semen. The ejaculated semen was divided into two portions: the first portion was centrifuged to separate the pellet of sperm from the seminal plasma and the second portion was cryopreserved. After thawing, the ejaculates were classified into three groups according to their post-thawed sperm motility: good (60.2 ± 1.7%), moderate (29.3 ± 2.0%) and poor (16.6 ± 2.2%) freezabilities. The expressions of GPX5 and FN1 in seminal plasma and TPI and ACRBP in sperm were determined using Western blot analysis. It was found that, for sperm proteins, the level of TPI was negatively correlated with the post-thawed total sperm motility (r = -0.38, P = 0.029). For seminal plasma proteins, the level of FN1 in the seminal plasma was positively correlated with the post-thawed total sperm motility (r = 0.37, P = 0.021) and progressive motility (r = 0.39, P = 0.016). The expression of GPX5 was not correlated with any of the frozen–thawed sperm qualities (P > 0.05). In conclusions, boar semen containing a high level of FN1 in seminal plasma has better freezability. Frozen–thawed sperm motility was positively correlated with the level of FN1 in boar seminal plasma and negatively correlated with TPI in boar spermatozoa.Ova studija provedena je u svrhu usporedbe ekspresije proteina sperme, tj. trioza-fosfat izomeraze (TPI) i akrozin-vezujućeg proteina (ACRBP) te proteina sjemene plazme, tj. glutation peroksidaze 5 (GPX5) i fibronektina 1 (FN1), u sjemenu nerasta s dobrom, umjerenom i lošom sposobnošću smrzavanja. Studija je provedena ustvrđivanjem sadržaja proteina u 32 uzorka sperme i 38 uzoraka sjemene plazme sjemena. Ejakulirano sjeme podijeljeno je u dva dijela: prvi dio je centrifugiran za odvajanje taloga sperme od sjemene plazme, a drugi je dio je krioprezerviran. Nakon odmrzavanja u skladu s pokretljivošću spermija nakon odmrzavanja ejakulati su klasificirani u tri skupine: dobra (60,2 ± 1,7%), umjerena (29,3 ± 2,0%) i loša (16,6 ± 2,2%) sposobnost smrzavanja. Ekspresije GPX5 i FN1 u sjemenoj plazmi te TPI i ACRBP u spermi ustvrđene su „Western blot“ analizom. Za proteine sperme je otkriveno da je razina TPI nakon odmrzavanja negativno povezana s ukupnom pokretljivošću sperme (r = -0,38, P = 0,029). Za proteine sjemene plazme, razina FN1 u sjemenoj plazmi nakon odmrzavanja pozitivno je povezana s ukupnom pokretljivošću sperme nakon odmrzavanja (r = 0,37, P = 0,021) i progresivnom pokretljivošću (r = 0,39, P = 0,016). Ekspresija GPX5 nije povezana ni sa kakvim kvalitetama smrznute pa odmrznute sperme (P > 0,05). Zaključno, sjeme nerasta koje sadrži visoku razinu FN1 u sjemenoj plazmi ima bolju sposobnost smrzavanja. Pokretljivost sperme koja je smrznuta pa odmrznuta pozitivno je povezana s razinom FN1 u sjemenoj plazmi nerasta, a negativno s razinom TPI u spermijima nerasta

Application of a novel rectangular filtering microfluidic device for microfilarial detection

The rectangular filtering microfluidic chip was invented using microfluidics device fabrication technology and can separate living microfilariae from blood samples without a syringe pump. The diagnostic results are highly effective. The device is based on the principle of separating millions of blood cells from microfilariae using a rectangular filter structure. It disperses fluid evenly into the flow-passage channel, and its rectangular filter structure is the key to success in reducing the pressure and separating blood cells from microfilariae effectively. The flow rate and blood cell concentration were optimized in our study. The chip is intended to be a point-of-care device that can reduce the use of superfluous instrumentation in the field. The technology is designed to be rapid, accurate, and easy-to-use for all users, especially those in remote areas

The Interaction of N-Acylhomoserine Lactone Quorum Sensing Signaling Molecules with Biological Membranes: Implications for Inter-Kingdom Signaling

The long chain N-acylhomoserine lactone (AHL) quorum sensing signal molecules released by Pseudomonas aeruginosa have long been known to elicit immunomodulatory effects through a process termed inter-kingdom signaling. However, to date very little is known regarding the exact mechanism of action of these compounds on their eukaryotic targets.The use of the membrane dipole fluorescent sensor di-8-ANEPPS to characterise the interactions of AHL quorum sensing signal molecules, N-(3-oxotetradecanoyl)-L-homoserine lactone (3-oxo-C14-HSL), N-(3-oxododecanoyl)homoserine-L-lactone (3-oxo-C12-HSL) and N-(3-oxodecanoyl) homoserine-L-lactone (3-oxo-C10 HSL) produced by Pseudomonas aeruginosa with model and cellular membranes is reported. The interactions of these AHLs with artificial membranes reveal that each of the compounds is capable of membrane interaction in the micromolar concentration range causing significant modulation of the membrane dipole potential. These interactions fit simple hyperbolic binding models with membrane affinity increasing with acyl chain length. Similar results were obtained with T-lymphocytes providing the evidence that AHLs are capable of direct interaction with the plasma membrane. 3-oxo-C12-HSL interacts with lymphocytes via a cooperative binding model therefore implying the existence of an AHL membrane receptor. The role of cholesterol in the interactions of AHLs with membranes, the significance of modulating cellular dipole potential for receptor conformation and the implications for immune modulation are discussed.Our observations support previous findings that increasing AHL lipophilicity increases the immunomodulatory activity of these quorum compounds, while providing evidence to suggest membrane interaction plays an important role in quorum sensing and implies a role for membrane microdomains in this process. Finally, our results suggest the existence of a eukaryotic membrane-located system that acts as an AHL receptor

Spatial characterisation of molecular interactions with cell membranes

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Serum protein profiles and C-reactive protein in natural canine filariasis

Background and Aim: Canine filariasis is caused by several species of filarial worms. The pathophysiological response to infection is mainly due to the filaria lifecycle. Laboratory detection methods to assess the pathological alterations characteristic of filariasis are needed urgently. Serum protein profiles and C-reactive protein (CRP) levels are used widely to diagnose several animal diseases. This study aimed to determine the serum protein profiles and CRP levels in dogs infected with Dirofilaria immitis or Brugia pahangi or both parasites. Materials and Methods: Blood samples were collected from 980 dogs presenting at animal hospitals and veterinary clinics in Bangkok and its vicinity. The presence of microfilaria in samples was determined using a buffy coat smear and staining with Wright–Giemsa. The sheathed and unsheathed microfilaria species were identified by acid phosphatase staining. Forty positive samples were tested. The serum protein profiles were identified by agarose gel electrophoresis. The CRP concentration was measured using a fluorescent immunoassay. Results: Albumin levels and albumin-to-globulin ratios were significantly lower, and total protein, β2 globulin, and γ globulin levels were significantly elevated in dogs infected with D. immitis and B. pahangi compared with reference values in normal dogs. The average CRP concentrations in dogs infected with D. immitis or B. pahangi were 69.9 and 12.9 mg/L, respectively. Conclusion: The total protein and γ globulin levels increased in canine filariasis compared with the normal reference range. The CRP concentration in dogs infected with D. immitis was extremely high, whereas that in dog infected with B. pahangi was normal

Microfilaria filtering microfluidic chip - Preliminary study

This paper provides a preliminary experiment of microfluidic filtering chip, designed to be used in an area that lacks of laboratory equipment. Without blood preparation involved sophisticated machines, the blood sample was used without rinsing or hemorrhaging before feeding into the chip. This chip contained with a wedge shape zone and detectable zone, which was in a cylindrical shape. The tests were performed with different sizes of inlet and length of the chip. From experimental results, the 5-μm micro capillaries allowed blood to flow through and was be able to filter a larger substance inside. However, appropriate blood dilution-ratio and filtering efficiency are needed to be clarified in the further experiment.</p