Periodontitis severity in obstructive sleep apnea patients.

OBJECTIVES: This cross-sectional study investigated the stages of periodontitis in obstructive sleep apnea (OSA) patients and risk factors associated with periodontitis severity among them. MATERIALS AND METHODS: A total of 194 patients underwent a polysomnography/polygraphy and were referred to periodontal examination. According to apnea-hypopnea index (AHI), patients were classified as mild OSA (AHI < 15) and moderate to severe OSA (AHI ≥ 15), whereas periodontitis severity was determined by the clinical attachment level (CAL) according to the recent Classification of Periodontal Diseases and Conditions. Patients were grouped into two categories: stages 1 and 2, and stages 3 and 4. RESULTS: Higher AHI values were reported in OSA patients exhibiting periodontitis stages 3 and 4 compared to OSA patients with periodontitis stages 1 and 2 (p = 0.043) and the non-periodontitis group (p = 0.044). A positive correlation was found between AHI and mean CAL (r = 0.215; p = 0.004), and between AHI and plaque scores (r = 0.292; p < 0.001). Following a multivariable regression analysis, AHI was a significant predictor of mean CAL (ß = 0.169; p = 0.031), explaining 16.4% of variability in mean CAL (adjusted R2 = 0.164; p < 0.001). Older patients had higher odds for an increased mean CAL (ß = 0.266; p = 0.001), as well as patients smoking or formerly smoking (ß = 0.305; p < 0.001) whereas visiting a dental medicine doctor once a year or more often was associated with a decreased mean CAL (ß = - 0.182; p = 0.02). CONCLUSIONS: OSA was associated with severe stages of periodontitis along with increased age, smoking, low frequency of dental visits, and poor oral hygiene. CLINICAL RELEVANCE: Screening for periodontitis is recommended for patients with more severe forms of OSA.

The construction of the Split Sleep Questionnaire on sleep habits during the COVID-19 pandemic in the general population.

AIM: To construct a single-format questionnaire on sleep habits and mood before and during the COVID-19 pandemic in the general population. METHODS: We constructed the Split Sleep Questionnaire (SSQ) after a literature search of sleep, mood, and lifestyle questionnaires, and after a group of sleep medicine experts proposed and assessed questionnaire items as relevant/irrelevant. The study was performed during 2021 in 326 respondents distributed equally in all age categories. Respondents filled out the SSQ, the Pittsburgh Sleep Quality Index (PSQI), and State Trait Anxiety Inventory (STAI), and kept a seven-day sleep diary. RESULTS: Cronbach alpha for Sleep Habits section was 0.819, and 0.89 for Mood section. Test-retest reliability ranged from 0.45 (P=0.036) for work-free day bedtime during the pandemic to 0.779 (P<0.001) for sleep latency before the pandemic. Workday and work-free day bedtime during the COVID-19 pandemic assessed with SSQ were comparable to the sleep diary assessment (P=0.632 and P=0.203, respectively), as was the workday waketime (P=0.139). Work-free day waketime was significantly later than assessed in sleep diary (8:19±1:52 vs 7:45±1:20; P<0.001). No difference in sleep latency was found between the SSQ and PSQI (P = 0.066). CONCLUSION: The SSQ provides a valid, reliable, and efficient screening tool for the assessment of sleep habits and associated factors in the general population during the COVID-19 pandemic.

The COVID-19 lockdown promotes changes in sleep habits in the Croatian general population.

AIM: To investigate the effects of the coronavirus disease 2019 (COVID-19) lockdown on sleep habits in the Croatian general population. METHODS: In this cross-sectional study, 1173 respondents from the general population (809 women) completed a self-report online questionnaire that gathered demographic data and data on sleep habits and mood changes before and during the COVID-19 lockdown. RESULTS: During the lockdown, bedtime (from 23:11±1:07 to 23:49±1:32 h, P<0.001) and waketime were delayed (from 6:51±1:09 to 7:49±1:40 h, P<0.001). Sleep latency increased from 10 (5-20) to 15 (10-30) minutes (P<0.001). Bedtime and waketime delays were more pronounced in women and respondents younger than 30. Compared with other age groups, respondents younger than 30 more frequently reported insomnia for the first time during the lockdown and had less frequent night-time awakenings (P<0.001), less common problems falling asleep (P<0.001), less frequently felt calm (P<0.001) and rested (P<0.001), but more frequently felt sadness (P<0.001) and fear (P=0.028). CONCLUSION: The effect of the lockdown on sleep needs to be better understood. Sleep hygiene education could serve a first-line lifestyle intervention for people in lockdown experiencing sleep disruption.

Comparison of Traditional and Industrial Sausages Baranjski Kulen and Kulenova Seka Using Comprehensive Proteome, Peptidome and Metabolome Techniques.

Research background: Baranjski kulen is one of the most popular fermented meat sausages originating from Croatia. It has protected geographical indication, and is traditionally produced in the Baranja region of Croatia. Kulenova seka is a fermented sausage very similar to baranjski kulen, but it has a different calibre and consequently, a shorter time of production. In recent decades, due to the high demand and popularity of these products, industrially produced baranjski kulen and kulenova seka have become available on the market. This work aims to identify specific characteristics of traditional and industrial sausages baranjski kulen and kulenova seka on proteome, peptidome and metabolome level, which could potentially lead to better optimization of the industrial production process in order to obtain an equivalent to the traditional product. Experimental approach: Protein profiles of baranjski kulen and kulenova seka (traditional and industrial) were analysed using two-dimensional gel electrophoresis followed by differential display analysis and protein identification by mass spectrometry. Peptidomic profile was analysed via liquid chromatography-tandem mass spectrometry. Furthermore, aroma profiles were investigated via headspace solid phase microextraction and gas chromatography-mass spectrometry. Results and conclusions: The major identified characteristics of each product were: industrial baranjski kulen: specific degradation of myosin-1 and titin, overabundance of stress-related proteins and increased phenylalanine degradation; traditional baranjski kulen: decreased concentration of phenylalanine and overabundance of fructose-bisphosphate aldolase A and carbonic anhydrase 3; industrial kulenova seka - specific myosin-4 and haemoglobin subunit alpha degradation process; traditional kulenova seka - overabundance of dihydropyrimidine dehydrogenase [NADP(+)] and myosin light chain 1/3, skeletal muscle isoform, degradation of albumin and myoglobin, decreased concentrations of almost all free amino acids and increased amounts of smoke-derived volatile compounds. Presented results showed that potential product type-specific quality markers for each sausage could be found. Novelty and scientific contribution: In this preliminary communication, the first insights into protein degradation processes and generation of peptides, free amino acids and aroma compounds of industrial and traditional baranjski kulen and kulenova seka are presented. Although further research is needed to draw general conclusions, the specific profile of proteins, peptides, amino acids and volatile compounds represents the first step in the industrial production of sausages that meet the characteristics of traditional flavour.

The effect of COVID-19 lockdown on lifestyle and mood in Croatian general population: a cross-sectional study.

AIM: To investigate the effect of the coronavirus 2019 (COVID-19) lockdown on lifestyle behaviors and mood changes in the Croatian general population. METHODS: During ten days of the COVID-19 lockdown in Croatia, 3027 respondents (70.3% female) from the general population completed an online, self-report questionnaire. Demographic data and data on lifestyle habits and mood changes before and during the COVID-19 lockdown were collected. RESULTS: A total of 95.64% of respondents reported to follow most or all restrictions, with female sex (P<0.001) and higher education level (P<0.001) being associated with higher restriction compliance. Women smoked an increased number of cigarettes (P<0.001). The proportion of respondents of both sexes who did not drink or drank 7 drinks per week or more increased (P<0.001). Women also reported lower frequency (P=0.001) and duration of physical exercise (P<0.001). In total, 30.7% of respondents gained weight, with female sex (OR, 2.726) and higher BMI (OR, 1.116; both P<0.001) being associated with an increased likelihood of gaining weight. Both men and women felt more frequently afraid (P<0.001), discouraged (P<0.001), and sad (P<0.001). CONCLUSION: Public health authorities should promote the adoption of healthy lifestyles in order to reduce long-term negative effects of the lockdown.

The effect of age and gender on cognitive and psychomotor abilities measured by computerized series tests: a cross-sectional study.

AIM: To assess age- and gender-associated differences in cognitive and psychomotor abilities measured by the Complex Reactionmeter Drenovac (CRD-series) tests. METHODS: This cross-sectional study, conducted between 2009 and 2019, enrolled 3420 participants (2012 women) in the age ranging from 18 to 88 years. The participants solved three CRD-series chronometric tests: discrimination of the light signal position (CRD311), complex psychomotor coordination (CRD411), and simple arithmetic operations (CRD11). We analyzed total test solving time (TTST), minimum single task solving time (MinT), number of errors, initial dissociation, and start, end, and total ballasts as measures of wasted time in the first half of the test, second half of the test, and total test time, respectively. RESULTS: Age was positively associated with MinT and TTST in all used tests (P<0.001), while initial dissociation, start ballast, and end ballast significantly increased with age (P<0.001). On the CRD11 test, men had shorter TTST than women (P=0.012), shorter start, end, and total ballasts (P<0.001), and made fewer errors than women (P<0.001). On the CRD311 test, women had shorter start (P=0.002), end, and total ballast (P<0.001) than men. On the CRD411 test, men performed better than women on all variables (P<0.001). CONCLUSION: Decreased cognitive and psychomotor abilities measured by the CRD-series tests were associated with advanced age. Men performed better than women on simple arithmetic and complex psychomotor coordination tests, whereas women lost less time on the test of light signal position discrimination.

Effects of CPAP therapy on cognitive and psychomotor performances in patients with severe obstructive sleep apnea: a prospective 1-year study.

STUDY OBJECTIVES: We prospectively investigated the effects of continuous positive airway pressure (CPAP) on long-term cognitive and psychomotor performances, and excessive daytime sleepiness in severe obstructive sleep apnea (OSA) patients. METHODS: A total of 40 patients were recruited and 23 patients with severe OSA fully completed the study protocol to investigate the effects of CPAP therapy on psychomotor performance at 1, 3, and 6 months and 1 year following initiation of the therapy. Psychomotor CRD-series tests measuring reaction times of light stimulus perception, solving simple arithmetic operations, and complex psychomotor limb coordination, were used in this study. The data collected following CPAP therapy were compared to baseline values prior to the CPAP treatment for each patient. RESULTS: All of the measured variables improved following CPAP treatment. However, the most pronounced effect was observed in improvement of reaction times to complex psychomotor limb coordination test (p < 0.05). Self-reported evaluation of excessive daytime sleepiness measured by Epworth Sleepiness Scale (ESS) showed significant decrease from 10.0 ± 1.1 before to 3.5 ± 0.5 (p < 0.001), after 1 year on CPAP therapy. CONCLUSIONS: The CPAP therapy improved cognitive and psychomotor performance on CRD-series tests with the most significant improvement observed in complex psychomotor limb coordination of severe OSA patients.

Phrenic long-term depression evoked by intermittent hypercapnia is modulated by serotonergic and adrenergic receptors in raphe nuclei.

Intermittent hypercapnia evokes prolonged depression of phrenic nerve activity (phrenic long-term depression, pLTD). This study was undertaken to investigate the role of 5-HT and α2-adrenergic receptors in the initiation of pLTD. Adult male urethane-anesthetized, vagotomized, paralyzed, and mechanically ventilated Sprague-Dawley rats were exposed to a protocol of acute intermittent hypercapnia (AIHc; 5 episodes of 15% CO2 in air, each episode lasting 3 min). The experimental group received microinjection of the selective 5-HT1A receptor agonist 8-hydroxy-2-(dipropylamino)tetralin hydrobromide (8-OH-DPAT), the broad-spectrum 5-HT antagonist methysergide, or the α2-adrenergic antagonist yohimbine, whereas the control group received microinjection of 0.9% saline into the caudal raphe region. Peak phrenic nerve activity (pPNA) and burst frequency ( f) were analyzed during baseline (T0), during 5 hypercapnic episodes (THc1-THc5), and at 15, 30, and 60 min after the end of the last hypercapnic episode. In the control group, pPNA decreased 60 min after the end of the last hypercapnic episode compared with baseline values, i.e., pLTD developed ( P = 0.023). In the 8-OH-DPAT group, pPNA significantly decreased at T15, T30, and T60 compared with baseline values, i.e., pLTD developed ( P = 0.01). In the methysergide and yohimbine groups, AIHc did not evoke significant changes of the pPNA at T15, T30, and T60 compared with baseline values. In conclusion, activation of 5-HT1A receptors accentuated induction of pLTD, whereas blockade of α2-adrenergic receptors prevented development of pLTD following AIHc in anesthetized rats. These results suggest that chemical modulation of 5-HT and α2-adrenergic receptors in raphe nuclei affects hypercapnia-induced pLTD, offering important insights in understanding the mechanisms involved in development of respiratory plasticity. NEW & NOTEWORTHY Hypercapnia is a concomitant feature of many breathing disorders, including obstructive sleep apnea. In this study, acute intermittent hypercapnia evoked development of phrenic long-term depression (pLTD) 60 min after the last hypercapnic episode that was preserved if the selective 5-HT1A receptor agonist 8-hydroxy-2-(dipropylamino)tetralin hydrobromide was microinjected in the caudal raphe region before the hypercapnic stimulus. This study highlights that both 5-HT and adrenergic receptor activation is needed for induction of pLTD in urethane-anesthetized rats following intermittent hypercapnia exposure.

Good and poor sleepers among OSA patients: sleep quality and overnight polysomnography findings.

Previous studies aimed to determine if Pittsburgh sleep quality index (PSQI) is a valid screening instrument for obstructive sleep apnea, indicating its disadvantages. However, the rationale of PSQI use in sleep clinics is not the screening, but the assessment of sleep quality itself. Therefore, the aims of this study were to investigate the sleep quality in obstructive sleep apnea patients and to identify the cutoff point for differentiation of "good" and "poor" sleepers among them. We constructed the Croatian version of PSQI and assessed its psychometric properties. The protocol of the study included the assessment of sleep quality in 130 obstructive sleep apnea patients and 75 healthy control subjects. All subjects completed the Croatian version of the PSQI, and the patients underwent overnight polysomnography screening. Obstructive sleep apnea patients had higher values of the global PSQI component score, indicating lower sleep quality, compared to a healthy control group (p < 0.001). The psychometric properties of PSQI scores in the prediction of normal sleep efficiency indicate that the cutoff score of 9.5 differentiates patients in total sleep time (p <  0.001), REM duration (p = 0.014), sleep efficiency (p = 0.001), time spent awake during sleep (p = 0.006), after sleep (p = 0.024), and after sleep onset (p = 0.001). In OSA patients, a PSQI cutoff score of 9.5 differentiated good and poor sleepers significantly in total sleep time, REM duration, time spent awake during sleep, and WASO time. Current findings enhance the interpretability of PSQI results in a population of OSA patients.

Intermittent hypercapnia-induced phrenic long-term depression is revealed after serotonin receptor blockade with methysergide in anaesthetized rats.

NEW FINDINGS: What is the central question of this study? Intermittent hypercapnia is a concomitant feature of breathing disorders. Hypercapnic stimuli evoke a form of respiratory plasticity known as phrenic long-term depression in experimental animals. This study was performed to investigate the putative role of serotonin receptors in the initiation of phrenic long-term depression in anaesthetized rats. What is the main finding and its importance? Phrenic nerve long-term depression was revealed in animals pretreated with the serotonin broad-spectrum antagonist, methysergide. This study highlights that serotonin receptors modulate respiratory plasticity evoked by acute intermittent hypercapnia in anaesthetized rats. This study was performed to test the hypothesis that intermittent hypercapnia can evoke a form of respiratory plasticity known as long-term depression of the phrenic nerve (pLTD) and that 5-HT receptors play a role in the initiation of pLTD. Adult male urethane-anaesthetized, vagotomized, paralysed, mechanically ventilated Sprague-Dawley rats were exposed to an acute intermittent hypercapnia protocol. One group received i.v. injection of the non-selective 5-HT receptor antagonist methysergide and another group received i.v. injection of the selective 5-HT1A receptor antagonist WAY-100635 20 min before exposure to intermittent hypercapnia. A control group received i.v. injection of saline. Peak phrenic nerve activity and respiratory rhythm parameters were analysed at baseline (T0), during each of five hypercapnic episodes, and 15, 30 and 60 min (T60) after the last hypercapnia. Intravenous injection of methysergide before exposure to acute intermittent hypercapnia induced development of amplitude pLTD at T60 (decreased by 46.1 ± 6.9%, P = 0.003). Conversely, in control and WAY-100635-pretreated animals, exposure to acute intermittent hypercapnia did not evoke amplitude pLTD. However, a long-term decrease in phrenic nerve frequency was evoked both in control (42 ± 4 breaths min(-1) at T0 versus 32 ± 5 breaths min(-1) at T60; P = 0.036) and in methysergide-pretreated animals (42 ± 2 breaths min(-1) at T0 versus 32 ± 3 breaths min(-1) at T60; P = 0.028). In WAY-100635 pretreated animals, frequency pLTD was prevented. These results suggest that 5-HT receptors modulate respiratory plasticity induced by acute intermittent hypercapnia in anaesthetized rats.

Daytime Sleepiness from Preschool Children's and Parents' Perspectives: Is There a Difference?

This cross-sectional study investigated the level of daytime sleepiness and sleep-related behaviors in preschool children and compared their self-evaluations with the evaluations of their parents. It was conducted in Split-Dalmatian County, Croatia, among 196 preschool children aged 6-7 years seen at regular medical examinations, accompanied by their parents, using the Epworth sleepiness scale for children and parents/caregivers. Compared to their child's reports, parents tended to underestimate their child's sleepiness while sitting in a classroom at school (p = 0.001) and overestimate their child's sleepiness when lying down to rest or nap in the afternoon (p < 0.001). Boys were sleepier while sitting in a classroom at school during the morning than girls (p = 0.032). As much as 48.2% of preschool children had their own cellphones/tablets. Boys used video games (p < 0.001) and cellphones/tablets more than girls did (p = 0.064). Parental estimation of children playing video games at bedtime was lower than the child's report (p < 0.001). Children who had a TV in their bedroom reported more daytime sleepiness (p = 0.049), and those who played video games at bedtime went to sleep later during the weekend (p = 0.024). Also, children owning cellphone/tablets had longer sleep latency during the weekend compared to children not owning a cellphone (p = 0.015). This study confirmed that parents tend to underestimate children's habits of playing video games at bedtime and children's sleepiness during morning classes. Preschool children who use electronic devices at bedtime more frequently have prolonged sleep latency. These findings provide further evidence of the effects of electronic media devices on preschoolers' sleep patterns and daytime sleepiness.

Comparison between enhanced MALDI in-source decay by ammonium persulfate and N- or C-terminal derivatization methods for detailed peptide structure determination.

Amino acid sequencing and more detailed structure elucidation analysis of peptides and small proteins is a very difficult task even if state-of-the-art mass spectrometry (MS) is employed. To make this task easier, chemical derivatization methods of the N terminus with 4-sulfophenyl-isothiocyanate (SPITC) or the C terminus with 2-methoxy-4,5-dihydro-1H-imidazole (Lys-tag) can enhance peptide fragmentation or fragment ionizability, via proton mobility/localization mechanisms making tandem MS (MS(2)) spectra more informative and less demanding for structural interpretation. Observed disadvantages related to both derivatization methods are sample- and time-consuming procedures and the increased number of reaction byproducts. A novel, sulfate radical in-source formation method of matrix-assisted laser desorption ionization (MALDI) MS based on chemically enhanced in-source decay (ISD) can be accomplished by simple addition of ammonium persulfate (APS) in the matrix solution. This method enables effective decomposition of peptide ions already in the first stage of MS analysis where a large number of fragment ions are produced. The resultant MALDI-ISD mass spectra (MS after APS → MALDI-ISD MS) are almost equivalent to conventional, collision-induced dissociation (CID) MS(2) spectra. These fragment ions are further subjected to the second stage of the MS, and consequently, MS(3) spectra are produced, which makes the sequence analysis more informative and complete (CID MS(2) is thus equivalent to CID MS(3)). Multiply stage MS after APS addition showed enhanced sensitivity, resolution, and mass accuracy compared to peptide derivatization (SPITC and Lys-tag) or conventional MS and MS(2) analyses and offered more detailed insight into peptide structure.

The effect of starvation stress on Lactobacillus brevis L62 protein profile determined by de novo sequencing in positive and negative mass spectrometry ion mode.

RATIONALE: We describe a novel negative chemically activated fragmentation/positive chemically activated fragmentation (CAF-/CAF+) technique for protein identification. The technique was used to investigate Lactobacillus brevis adaptation to nutrient deprivation. METHODS: The CAF-/CAF+ method enables de novo sequencing of derivate peptides with negative and positive ion mode matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry (MS/MS). Peptide sequences obtained from MS/MS spectra were matched against the National Center for Biotechnology Information (NCBI) non-redundant (nr) database and confirmed by the mass spectrometry data of elucidated peptide mass sequences derived from the annotated genome. This improved protein identification method highlighted 36 differentially expressed proteins in the proteome of L. brevis after 75 days of starvation. RESULTS: The results revealed the key differences in the metabolic pathways that are responsible for the survival of L. brevis in a hostile environment. Proteomics analysis demonstrated that numerous proteins engaged in glucose and amino-acid catabolizing pathways, glycerolipid metabolizing pathways, and stress-response mechanisms are differentially expressed after long-term starvation. Amino acid and proteomics analysis indicated that starved L. brevis metabolized arginine, glycine, and histidine from dead cells as alternative nutrient sources. The production of lactic acid also varied between the parent cells and the starved cells. CONCLUSIONS: Differentially expressed proteins identified exclusively by peptide sequence reading provided promising results for CAF-/CAF+ implementation in a standard proteomics workflow (e.g., biomarker and mutation discovery and biotyping). The practical performance of a reliable de novo sequencing technique in routine proteomics analysis is emphasized in this article.

The Associations between Results in Different Domains of Cognitive and Psychomotor Abilities Measured in Medical Students.

We aimed to investigate the associations between intelligence quotient test scores obtained using the Raven's Advanced Progressive Matrices (APM) and psychomotor testing using the Complex Reactionmeter Drenovac (CRD) test battery, while taking into account previous theoretical approaches recognizing intelligent behavior as the cumulative result of a general biological speed factor reflected in the reaction time for perceptual detections and motor decisions. A total of 224 medical students at the University of Split School of Medicine were recruited. Their IQ scores were assessed using Raven's APM, while the computerized tests of CRD-series were used for testing the reaction time of perception to visual stimulus (CRD311), psychomotor limbs coordination task (CRD411), and solving simple arithmetic operations (CRD11). The total test-solving (TTST) and the minimum single-task-solving (MinT) times were analyzed. On the CRD11 test, task-solving times were shorter in students with higher APM scores (r = -0.48 for TTST and r = -0.44 for MinT; p < 0.001 for both). Negative associations between task-solving times and APM scores were reported on CRD311 (r = -0.30 for TTST and r = -0.33 for MinT, p < 0.001 for both). Negative associations between task-solving times in CRD411 and APM scores (r = -0.40 for TTST and r = -0.30 for MinT, p < 0.001 for both) were found. Faster reaction time in psychomotor limbs coordination tasks, the reaction time of perception to visual stimulus, and the reaction time of solving simple arithmetic operations were associated with a higher APM score in medical students, indicating the importance of mental speed in intelligence test performance. However, executive system functions, such as attention, planning, and goal weighting, might also impact cognitive abilities and should be considered in future research.

Sleep and Lifestyle Habits of Medical and Non-Medical Students during the COVID-19 Lockdown.

It has been shown that the measures of social distancing and lockdown might have had negative effects on the physical and mental health of the population. We aim to investigate the sleep and lifestyle habits as well as the mood of Croatian medical (MS) and non-medical students (NMS) during the COVID-19 lockdown. The cross-sectional study included 1163 students (21.6% male), whose lifestyle and sleep habits and mood before and during the lockdown were assessed with an online questionnaire. The shift towards later bedtimes was more pronounced among NMS (~65 min) compared to MS (~38 min), while the shift toward later wake-up times was similar in both MS (~111 min) and NMS (~112 min). All students reported more frequent difficulty in falling asleep, night-time awakenings and insomnia (p < 0.001) during lockdown. A higher proportion of MS reported being less tired and less anxious during lockdown compared to pre-lockdown (p < 0.001). Both student groups experienced unpleasant moods and were less content during lockdown compared to the pre-lockdown period (p < 0.001). Our results emphasize the need for the promotion of healthy habits in the youth population. However, the co-appearance of prolonged and delayed sleep times along with decreased tiredness and anxiety among MS during lockdown reveals their significant workload during pre-lockdown and that even subtle changes in their day schedule might contribute to the well-being of MS.

Not-So-Sweet Dreams: Plasma and IgG N-Glycome in the Severe Form of the Obstructive Sleep Apnea.

Obstructive sleep apnea (OSA) is a prevalent disease associated with increased risk for cardiovascular and metabolic diseases and shortened lifespan. The aim of this study was to explore the possibility of using N-glycome as a biomarker for the severe form of OSA. Seventy subjects who underwent a whole-night polysomnography/polygraphy and had apnea-hypopnea index (AHI) over 30 were compared to 23 controls (AHI under 5). Plasma samples were used to extract 39 glycan peaks using ultra-high-performance liquid chromatography (UPLC) and 27 IgG peaks using capillary gel electrophoresis (CGE). We also measured glycan age, a molecular proxy for biological aging. Three plasma and one IgG peaks were significant in a multivariate model controlling for the effects of age, sex, and body mass index. These included decreased GP24 (disialylated triantennary glycans as major structure) and GP28 (trigalactosylated, triantennary, disialylated, and trisialylated glycans), and increased GP32 (trisialylated triantennary glycan). Only one IgG glycan peak was significantly increased (P26), which contains biantennary digalactosylated glycans with core fucose. Patients with severe OSA exhibited accelerated biological aging, with a median of 6.9 years more than their chronological age (p < 0.001). Plasma N-glycome can be used as a biomarker for severe OSA.

The disulfide bond of an RGD4C motif inserted within the Hi loop of the adenovirus type 5 fiber protein is critical for retargeting to αv -integrins.

BACKGROUND: The α(v) -integrin binding motif RGD4C (CDCRGDCFC) has been used extensively to circumvent inefficient adenovirus type 5 (Ad5) transduction of cells expressing low levels of the coxsackie and adenovirus receptor. However, until now, it has been unclear whether disulfide bonds in the RGD4C motif influence the retargeting potential of RGD4C-modified Ad5. METHODS: Replication deficient Ad5 bearing wild-type fiber (Ad5wt) or RGD4G, RGD4C and RGD2C2G insertions within the HI loop of the fiber protein (Ad5RGD4G, Ad5RGD4C and Ad5RGD2C2G, respectively) were used to transduce a panel of cancer cell lines, with or without previous treatment of these Ad5s with the reducing agent dithiothreitol (DTT). In parallel, native and DTT-treated fiber proteins isolated from purified Ad5RGD4C were compared by mass spectrometry. RESULTS: Ad5RGD4C transduced all studied cell lines much more efficiently than Ad5wt, whereas Ad5RGD4G transduced cells only slightly more efficiently than Ad5wt. DTT treatment had no effect on cell transduction by wild-type Ad5wt and Ad5RGD4G but abolished the increased transduction efficacy of Ad5RGD4C in a dose-dependent manner. The mass spectra of native and DTT-reduced tryptic digests of the Ad5RGD4C fiber protein are consistent with the presence of a C(547) -C(549) linkage in the C(547) DC(549) RGDC(553) FC(555) motif. Finally, the high transduction efficacy of Ad5RGD4C is conserved in Ad5RGD2C2G. CONCLUSIONS: We provide genetic and biochemical data strongly suggesting that cysteines C(547) and C(549) from the C(547) DC(549) RGDC(553) FC(555) motif inserted in the HI loop of the Ad5 fiber form a single disulfide bond, with this disulfide bond being crucial for Ad5RGD4C retargeting to av-integrins.

Acute intermittent hypoxia induces phrenic long-term facilitation which is modulated by 5-HT1A receptor in the caudal raphe region of the rat.

Obstructive sleep apnoea (OSA) is characterized by periods of upper airway collapse accompanied by repeated episodes of hypoxia. In experimental animals repeated bouts of hypoxia may evoke sustained augmentation of phrenic nerve activity, known as phrenic long-term facilitation (pLTF). This form of physiological compensation might contribute to stable breathing, minimizing the occurrence of apnoeas and/or hypopnoeas during sleep in patients with OSA. Serotonin (5-HT) has been shown to modulate respiratory neuronal activity, possibly via projections originating in the raphe nuclei. Our model focuses on the effects of 5-HT1A receptors blockade by selective antagonist WAY-100635 into the caudal raphe region on phrenic long-term facilitation after exposure to acute intermittent hypoxia (AIH) episodes. Adult, male, urethane-anaesthetized, vagotomized, paralyzed and mechanically ventilated Sprague-Dawley rats were exposed to AIH protocol. Experimental group received microinjection of WAY-100635 into the caudal raphe nucleus, whereas the control group received saline into the same site. Peak phrenic nerve activity and respiratory rhythm parameters were analysed during five hypoxic episodes, as well as at 15, 30 and 60 min after the end of hypoxias. In the control group, 1 h post-hypoxia pLTF was developed. Microinjections of selective 5-HT1A receptor antagonist WAY-100635 into the raphe nuclei prior to the AIH protocol prevented induction of pLTF. These results suggest that 5-HT1A receptor activation at supraspinal level is important for induction of pLTF, which is suggested to be an important respiratory neuroplasticity model in animal studies that possibly correlates with OSA in humans.

The evaluation of the Croatian version of the Epworth sleepiness scale and STOP questionnaire as screening tools for obstructive sleep apnea syndrome.

PURPOSE: Growing awareness of obstructive sleep apnea syndrome (OSAS) has increased the need for concise and reliable screening tools. The Epworth sleepiness scale (ESS) has been validated in numerous languages and ethnic groups, since it was originally designed for the English-speaking population. The STOP questionnaire was developed as a novel OSAS screening tool in surgical patients, but has not been validated in the general population. The present study was undertaken to provide reliable and validated ESS in the Croatian language and to evaluate the ESS and STOP as screening instruments for OSAS. METHODS: The Croatian version of ESS and STOP questionnaire was administered to 217 patients referred to the Split Sleep Medicine Center and 208 healthy control subjects. Test-retest reliability was investigated in 20 healthy subjects. RESULTS: The ESS score was significantly higher for the patients referred to the Split Sleep Medicine Center compared to the control group (8.2 ± 5.0 vs. 5.9 ± 3.8, p < 0.001). Cronbach's alpha coefficient for the ESS Croatian version was 0.84 indicating an excellent internal consistency. Reproducibility revealed no significant difference in each item or in the total ESS scores. Receiver operating curve of the ESS for identification of cases with AHI >5/h was 0.64, and for the STOP questionnaire, it was 0.84. CONCLUSIONS: Both ESS and STOP questionnaires successfully distinguished healthy subjects from subjects with OSAS. The STOP questionnaire had better probability to correctly predict high-risk patients for OSAS compared to ESS. We propose that the STOP questionnaire could be used as an easy-to-use and accurate screening tool in identification of patients with risk for OSAS in the general population, but it has not been tested in the Croatian population yet.

Salivary parameters and periodontal inflammation in obstructive sleep apnoea patients.

The aim of this cross-sectional study was to objectively assess the salivary flow rate and composition and periodontal inflammation in obstructive sleep apnoea (OSA) patients. The subjects, who underwent whole-night polysomnography or polygraphy, were referred for saliva sampling and periodontal examination. According to the severity of OSA based on the Apnoea Hypopnea Index (AHI) value, the subjects were classified into groups: no OSA (AHI < 5; N = 17), mild to moderate OSA (AHI 5-29.9; N = 109), and severe OSA (AHI > 30; N = 79). Salivary flow rate, pH, salivary electrolytes, and cortisol were measured from collected saliva samples. Periodontal examination included assessment of the number of teeth, dental plaque, bleeding on probing and periodontal measurements: gingival recession, probing pocket depth, clinical attachment level (CAL) and periodontal inflamed surface area (PISA) score. There were no significant differences in salivary flow rate, salivary pH, salivary electrolyte concentrations or electrolyte ratios among the groups classified according to the severity of OSA. However, subjects without OSA had higher salivary cortisol concentrations than OSA groups (p < 0.001). Increased plaque scores were associated with a higher AHI (r = 0.26; p = 0.003). According to the salivary flow rate, subjects with hyposalivation and reduced salivation had higher concentrations of salivary electrolytes and lower salivary pH than subjects with normal salivation. Subjects with hyposalivation had an increased Mg/PO4 ratio (p < 0.001) and a reduced Ca/Mg ratio (p < 0.001). Furthermore, subjects with severe OSA tended to have higher CALs and plaque volumes. In conclusion, under pathological conditions, such as OSA, multiple interactions might impact salivary flow and electrolyte composition. Complex interrelationships might affect the integrity of oral health, especially considering OSA severity, inflammation, concomitant diseases and medications.