Advanced Fluorescence Microscopy Techniques-FRAP, FLIP, FLAP, FRET and FLIM

Fluorescence microscopy provides an efficient and unique approach to study fixed and living cells because of its versatility, specificity, and high sensitivity. Fluorescence microscopes can both detect the fluorescence emitted from labeled molecules in biological samples as images or photometric data from which intensities and emission spectra can be deduced. By exploiting the characteristics of fluorescence, various techniques have been developed that enable the visualization and analysis of complex dynamic events in cells, organelles, and sub-organelle components within the biological specimen. The techniques described here are fluorescence recovery after photobleaching (FRAP), the related fluorescence loss in photobleaching (FLIP), fluorescence localization after photobleaching (FLAP), Forster or fluorescence resonance energy transfer (FRET) and the different ways how to measure FRET, such as acceptor bleaching, sensitized emission, polarization anisotropy, and fluorescence lifetime imaging microscopy (FLIM). First, a brief introduction into the mechanisms underlying fluorescence as a physical phenomenon and fluorescence, confocal, and multiphoton microscopy is given. Subsequently, these advanced microscopy techniques are introduced in more detail, with a description of how these techniques are performed, what needs to be considered, and what practical advantages they can bring to cell biological research

Evidence for autonomic function and its influencing factors in subjects with COPD: a systematic review

Cardiovascular autonomic neuropathy is one of the factors implicated in the high morbidity and mortality rate in patients with COPD. Thus, several studies and nonsystematic reviews have increasingly reported autonomic function impairment in these subjects. For a better understanding, this systematic review was performed to evaluate not only the evidence for autonomic function impairment, but also factors influencing it. The results of the studies reviewed showed a strong level of evidence to support the impairment of heart rate variability in the time domain. A similar evidence level was also found to support impairment in baroreceptor sensitivity and muscle sympathetic nerve activity. Furthermore, this review identified physical activity level, muscle function, and circadian rhythm as the major influencing factors (strong evidence) of autonomic function in subjects with COPD. However, no definite conclusion could be reached for factors such as dyspnea, anxiety, body composition, pulmonary function, age, breathing frequency, ventilatory effort, quality of life, and disease severity due to limited, conflicting, or lack of existing evidence. The results of this review highlight relevant clinical messages for clinicians and other health-care providers regarding the role autonomic function can play as an important physiological marker for prognostication and stratification. Hence, autonomic function outcomes should be identified and considered during management of patients with COPD. Moreover, this review can serve as basis for future research aimed at assessing the interventions for autonomic function abnormalities in these patients

Temporal Changes in Blood Biomarkers Associated with Sleep Apnea Severity: A Retrospective Cohort Study in Finland

Obstruktiivinen uniapnea on yleinen unihäiriö, johon liittyy unenaikaisia hengityskatkoksia, jotka usein johtavat happisaturaation laskuun ja unen häiriintymiseen. Uniapnean tiedetään vaikuttavan moniin fysiologisiin parametreihin, kuten hematologisiin ja lipidiarvoihin. Tämän tutkimuksen tavoitteena on tutkia CPAP-hoidon vaikutuksia uniapneapotilaiden laboratorioarvoihin. Toteutimme retrospektiivisen tutkimuksen, jossa käytimme Helsingin ja Uudenmaan sairaanhoitopiirin (HUS) aineistoa, joka kattoi 30 722 vuosina 2005–2020 hoidettua aikuista uniapneapotilasta. Tutkimuksessa hyödynsimme sanahakua, jonka avulla etsimme aineistosta tietoa potilaiden apnea-hypopnea-indeksistä (AHI), CPAP-hoidon käytöstä ja tunnistaaksemme hoidosta kieltäytyneet potilaat. Hematologiset ja metaboliset laboratorioarvot kerättiin kolme vuotta ennen potilaan ensimmäistä uniapneadiagnoosia ja kolme vuotta diagnoosin jälkeen. Kovarianssianalyysiä käytettiin vertailemaan parametrien vaihtelua uniapnean eri vaikeusasteiden välillä vakioituna iällä, sukupuolella ja BMI:llä. Parittaista T-testiä käytettiin analysoimaan eroja ennen ja jälkeen uniapneadiagnoosin. Uniapnean eri vaikeusasteita esiintyi aineistossa seuraavasti: 14,8 % lieväasteisia, 32,6 % keskivaikea-asteisia ja 52,6 % vaikea-asteisia. Keskimääräinen diagnosointi-ikä oli 55,0 ja keskimääräinen BMI 32,4. Kliinisesti merkittävimmät muutokset havaittiin veren lipidiarvoissa kolesteroli-, HDL- ja LDL-tasojen kohentumisena (P < 0,05) mitattuna ennen CPAP-hoidon aloitusta ja hoidon aloittamisen jälkeen. Glukoositasot sen sijaan nousivat seurantajakson aikana. Vastaavasti hematokriitti ja hemoglobiini laskivat merkitsevästi CPAP-hoidon aloituksen jälkeen. Sukupuolikohtaisissa analyyseissä merkittävää parantumista kolesteroli-, HDL- ja LDL-tasoissa havaittiin molemmilla sukupuolilla. Triglyseriditasot parantuivat miespotilailla, mutta naispotilailla ne päinvastoin nousivat seurantajakson aikana. CPAP-hoito kohentaa kolesteroli-, HDL- ja LDL-tasoja merkittävästi molemmilla sukupuolilla sekä alentaa hematokriitti- ja hemoglobiinitasoja. Tämä tutkimus korostaa uniapnean systeemisiä vaikutuksia ja alleviivaa hematologisen ja lipidiprofiilin arvioinnin tärkeyttä uniapnean hallinnassa.Obstructive sleep apnea (OSA) is a common sleep disorder associated with breathing interruptions during sleep, often leading to oxygen level drops and sleep disturbances. OSA is known to impact various physiological parameters, including hematological and lipid profiles. This study aims to investigate the effect of continuous positive airway pressure (CPAP) therapy on laboratory values in patients with OSA. We conducted a retrospective study using data from Finland's largest hospital district, including 30,722 adult OSA patients treated between 2005 and 2020. We implemented a text search algorithm within the patient chart data to extract the apnea-hypopnea index (AHI) and the usage of CPAP therapy, along with identifying patients who had declined treatment. Hematological and metabolic laboratory values were collected three years before and after the first OSA diagnosis. Analysis of covariance (ANCOVA) was employed to compare parameter variations across severity levels, adjusted for age, sex, and BMI. T-test for repeated measurements was used to analyze the differences between three years prior and three years after the first OSA diagnosis. Our study of 30,722 OSA patients showed varying severity levels in OSA patients: 14.8% mild, 32.6% moderate, and 52.6% severe, with an average diagnosis age of 55.0 years and a mean BMI of 32.4. The most clinically significant changes were observed in lipid profile markers, with improvements in cholesterol, HDL, and LDL levels (P < 0.05), measured before CPAP treatment initiation and after the treatment began. Conversely, glucose levels increased during the follow-up period. Similarly, hematocrit and hemoglobin decreased significantly after initiation of the CPAP treatment. In sex-specific analyses, significant improvements in cholesterol, HDL, and LDL levels were found in both sexes. Triglyceride levels improved in male patients, in contrast to female participants, whose triglyceride levels increased during the follow-up period. CPAP therapy significantly improves cholesterol, HDL, and LDL levels in both sexes and reduces hematocrit and hemoglobin levels. This study highlights the systemic effects of OSA and underscores the importance of evaluating hematological and lipid profiles in OSA management

The quality of duck eggs at different lengths of storage by the addition of Tilapia fish oil and noni fruit extract to the feed

This study aimed to determine the effect of the addition of tilapia oil and noni fruit extract to duck feed on egg quality at different storage times. A total of 80 duck eggs were used as research material using the factorial complete randomized design (CRD) method. In this study, there were two types of treatments: factor A was the diet, which included adding tilapia fish oil and noni fruit extract. The treatments were A1 (no added fish oil or noni fruit extract), A2 (0.5 percent fish oil and 0.75 percent noni fruit extract), A3 (0.75% fish oil and 0.75% noni fruit extract), and A4 (1% fish oil and 1% noni fruit extract). Factor B was the length of time the eggs were stored, with B1 (no storage), B2 (storage for 7 days), B3 (storage for 14 days), and B4 (stored for 21 days). The results showed that the Haugh unit value, the egg white index value, and the percentage of egg weight shrinkage in the storage treatment were significantly different at each length of storage time; the yolk index value and the yolk pH value were significantly different at day 14 and day 21 storage; and the egg white pH value at day 0 storage was significantly different from days 7, 14, and 21 storage. This study concludes there was no interaction between the two treatment factors and the egg quality at different storage times

The quality of duck eggs at different lengths of storage by the addition of Tilapia fish oil and noni fruit extract to the feed

This study aimed to determine the effect of the addition of tilapia oil and noni fruit extract to duck feed on egg quality at different storage times. A total of 80 duck eggs were used as research material using the factorial complete randomized design (CRD) method. In this study, there were two types of treatments: factor A was the diet, which included adding tilapia fish oil and noni fruit extract. The treatments were A1 (no added fish oil or noni fruit extract), A2 (0.5 percent fish oil and 0.75 percent noni fruit extract), A3 (0.75% fish oil and 0.75% noni fruit extract), and A4 (1% fish oil and 1% noni fruit extract). Factor B was the length of time the eggs were stored, with B1 (no storage), B2 (storage for 7 days), B3 (storage for 14 days), and B4 (stored for 21 days). The results showed that the Haugh unit value, the egg white index value, and the percentage of egg weight shrinkage in the storage treatment were significantly different at each length of storage time; the yolk index value and the yolk pH value were significantly different at day 14 and day 21 storage; and the egg white pH value at day 0 storage was significantly different from days 7, 14, and 21 storage. This study concludes there was no interaction between the two treatment factors and the egg quality at different storage times