Anti-Carbamylated Protein Antibodies in ACPA-Negative and ACPA-Positive Patients with Rheumatoid Arthritis.

The objective of this study was to assess the level of antibodies to carbamylated proteins and analyze the clinical and immunological associations in patients with ACPA-negative and ACPA-positive variants of rheumatoid arthritis. MATERIALS AND METHODS: . The study involved 150 patients with a reliable diagnosis of rheumatoid arthritis and 25 patients as healthy controls. Depending on ACPA values, two groups of patients were recruited: ACPA-positive (n = 75) and ACPA-negative (n = 75). RA activity was assessed by the DAS28 index. Determination of antibodies to carbamylated proteins was performed by enzyme-linked immunosorbent assay (BlueGene Biotech, China). Quantitative determination of ACPA in serum was performed by enzyme immunoassay using a commercial reagent kit (AxisShield, UK; upper limit of normal 5.0 U/mL; Orgentec, Germany; upper limit of normal 20.0 U/mL). RESULTS AND DISCUSSION: . Median anti-CarP in patients with RA was 126.2 [100.83; 157.41] ng/mL and was statistically significantly higher (p < 0.001) than in healthy controls (88.89 [70.53; 107.75] ng/mL). Among all patients with RA, 50 (33.3%) were anti-Carp-positive (22 (29.3%) in the ACPA(+) group and 28 (37.3%) in the ACPA(-) group), and one (2%) volunteer from healthy controls was anti-CarP(+) (p = 0.002). In ROC analysis performed to assess the diagnostic significance of anti-CarP for RA for all patients with RA, the area under the curve was 0.783 ± 0.047 with 95% CI: 0.691-0.874 (p < 0.001), with a cut-off point of 143 ng/mL, specificity 96%, sensitivity 36.7%. In the ACPA(+) RA group, the erosion count was statistically significantly higher (p = 0.044) in anti-CarP(+) patients than in anti-CarP(-) patients. A weak direct correlation between anti-CarP and DAS28 was found in the ACPA(-) RA group. CONCLUSIONS: . We studied the predictive value of anti-CarP as an auxiliary biomarker in ACPA(+) and ACPA(-) subtypes of RA. ACPA(+) anti-CarP(+) patients have a more "erosive" subtype of the disease than ACPA(+) anti-CarP(-) patients. In ACPA(-) patients, anti-CarP helps to identify a more erosive subtype of the disease, and among ACPA(-) patients it helps to reduce the proportion of seronegative patients. Further studies are required to determine the optimal standards for the laboratory diagnosis of anti-CarP and to clarify the diagnostic potential of these ABs as part of the differential diagnosis of arthritis in other rheumatic diseases.

Association between Ultradian Rhythms of Body Temperature in Small Mammals and Earth's Crust Stress.

Association was assessed between the data harvested by a long-baseline laser interference deformograph and the dynamics of body temperature (BT) in hamsters deprived of natural daily light-darkness changes. The power spectral data revealed the positive correlation between simultaneous time series of hamster BT and the Earth's crust deformation (ECD). The superposed epoch analysis established an association between abrupt upstrokes of hamster BT and ECD increments. Thus, the direct relationships between BT dynamics (reflecting predominance of sympathetic part of autonomic nervous system) and ECD (according to long-baseline laser interference deformography) were established. The study observed synchronization of the free-running circadian rhythm of hamster BT with the tidal stress in Earth's lithosphere. Further studies are needed to find the physical factor underlying the revealed relationships.

[Bone mineral density in women with rheumatoid arthritis: A link between immune and biochemical markers].

AIM: To study the association of bone mineral density (BMD) with serum biochemical and immunological markers in postmenopausal women with rheumatoid arthritis (RA). MATERIALS AND METHODS: The study included 173 women with RA (age 61.0 [56.0; 66.0] years). A survey, dual-energy X-ray absorptiometry to measure the BMD of the lumbar spine (LI-LIV), femoral neck (FN) and total hip (TH), routine blood chemistry, measurement of C-reactive protein (CRP), rheumatoid factor, cyclic citrullinated peptide antibodies (CCPA), parathyroid hormone (PTH), vitamin D3, myostatin, follistatin, interleukin-6 (IL-6), IL-6 receptors, insulin-like growth factor 1, adiponectin, leptin, fibroblast growth factor 23, and tumor necrosis factor SF12 were performed. RESULTS: PTH (ß=-0.22, -0.35 and -0.30 for LI-LIV, FN and TH, respectively), CRP (ß=-0.18, 0.23 and -0.22 for LI-LIV, FN and TH, respectively) and leptin (ß=0.35, 0.32 and 0.42 for LI-LIV, FN and TH, respectively) were shown a significant association with BMD in all sites of measurement. It was independent of age, body mass index and postmenopause duration. Associations were also found between adiponectin and BMD of LI-LIV and TH (ß=-0.36 and -0.28, respectively), CCPA and BMD of FN and TH (ß=-0.21, -0.24, respectively) and IL-6 and BMD of FN (ß=0.37). CONCLUSION: The study of biochemical and immunological markers in women with RA demonstrated that CRP, CCPA, PTH, IL-6, adiponectin, and leptin influenced BMD.

Influence of Melatonin on Maturation of Male Germ Cells.

We studied the influence of epiphysectomy and administration of melatonin to epiphysectomized outbred white rats on the level and daily dynamics of damage to the genetic material of developing male germ cells. Epiphysectomy leads to an increase in the level of damage in the DNA structure and the disappearance of the circadian rhythm of the activity of repair enzyme PARP-1 and the apoptosis-inducing enzyme caspase-3. The administration of melatonin to animals after epiphysectomy reduces the level of DNA damage, restores the circadian rhythm of activity of PARP-1 and caspase-3. These fundings suggest that melatonin can indirectly protect DNA of maturing male gametes.

Intensity of 40K Decay Fluctuations an Indicator of an Environmental Factor Determining Infradian Rhythms of Mouse Motor Activity, Circulating Cortisol in Rabbits, and Proliferation of L-929 Cultured Cells.

The multi-day dynamics of the intensity of 40K decay fluctuations was compared with motor activity of laboratory mice, circulating cortisol in rabbits, and proliferation of fibroblast-like cultured cells L-929. A positive correlation was established between the intensity of decay fluctuations on the one hand, and total daily motor activity in mice and plasma cortisol level in rabbits, on the other hand. In addition, a negative correlation was observed between 40K decay fluctuations and proliferative activity of the cultured cells. Interestingly, these close correlations were observed not only against the background 4-day rhythm, but also in the time when any rhythmicity was absent. Thus, the intensity of 40K decay fluctuations is an indicator of a biotropic environmental factor that augments motor activity and raises the circulating cortisol in animals and down-regulates proliferation of cultured cells.

Effect of Heliogeophysical and Atmospheric Factors on the Degree of Synchronization of Ultradian Rhythms of Body Temperature in Mice.

In a long-term (8 months) study, we examined the degree of synchronization of ultradian body temperature oscillations of two isolated groups of mice kept under constant dim illumination. In most cases, the periods of increased activity accompanied by rapid elevation of body temperature coincided in these groups of mice, but in some days, no significant synchronization between the examined parameters was observed. Analysis of the effects of environmental factors on the degree of synchronization of ultradian rhythms in mice revealed association of this parameter with the dynamics of atmospheric pressure (AtmP) and to a lesser extent with the vertical component of interplanetary magnetic field Bz. The loss in synchronicity of ultradian rhythms of mouse activity occurred after a rapid drop of AtmP or during pronounced negative Bz. Therefore, these factors can be viewed as desynchronizers of the biological ultradian rhythms.

Analysis of Association of Ultradian Body Temperature Rhythms in Animals with Intensity of Fluctuations of Radioactive Decay of Natural 40K Isotope.

The study monitored the long-term body temperature (BT) oscillations of C57BL/6 mice and outbred starlings (Sturnus vulgaris) to compare them with fluctuation in decay rate of radioactive natural 40K isotope. The spectrum analysis revealed simultaneous changes of the predominant periods in BT spectra of the animals and those in fluctuation in 40K decay rate. A positive correlation was established between BT dynamics and fluctuation in decay rate. The superposed epoch analysis revealed predominant coincidence of the moments of BT and fluctuation in 40K decay rate. The novel data indicate association between BT ultradian rhythms with quasirhythmic variations of fluctuation in 40K decay rate.

Fluctuations in the Rate of Radioactive Decay of the Natural Potassium-40 Isotope Can Potentially Affect the Periodic "Spontaneous" Activity of Animals.

Body temperature rhythms of C57Bl/6 laboratory mice and common greenfinches (Chloris chloris) and feeding periods of common starlings (Sturnus vulgaris) were compared with the intensity of fluctuations in 40К radioactive decay. Body temperature changes in greenfinches and mice were found to positively correlate with the intensity of fluctuations in 40K radioactive decay. Superposed epoch analysis showed that an increase in mouse body temperature, which reflects the start of the active phase in the sleep-wake cycle, and food intake in starlings coincide with an increase in the intensity of 40K radioactive decay. Thus, animal activity in the ultradian range of periods may be related to external quasi-rhythmic physical influences, rather than being determined only by endogenous processes. Given the extremely low dose of natural 40К exposure, a factor responsible for radioactivity fluctuations may act as a biotropic factor.

Ultradian Near 2-4-h Rhythms of Body Temperature in Laboratory Rodents Depend on External Environmental Heliogeophysical Factor Reflected by Neutron Monitor Count Rate.

The study collated oscillations of body temperature of С57BL/6 mice and Roborovski hamsters (Phodopus roborovskii) in the period range of 100-240 min and fluctuations of the secondary cosmic rays reflected by neutron count rate of a neutron monitor at the Earth's surface. Cross-correlation of body temperature and neutron count rate data revealed their strong association only for simultaneous measurements. Collation of the phase of neutron count rate fluctuations with maxima of body temperature oscillations revealed the coincidence of the phases of both processes. There was a temporary connection between the periods of body temperature elevation and neutron count rate rise. Thus, not only the spectral parameters of biological and physical processes, but also their phases coincided, which corroborates the hypothesis on stimulating effect of increasing neutron count rate on activity of the animals.

Association of Infradian Rhythms of Motor Activity, Concentration of Glucocorticoid Hormones, and One-Minute-Step Oscillations of Body Temperature with Intensity of Fluctuations of Secondary Cosmic Rays.

The study compared the daily mean intensity of one-minute-step fluctuations in intensity of the secondary cosmic rays reflected by neutron count rate and dynamics of body temperature, motor activity, as well as concentration of glucocorticoid hormones in birds and rodents. A positive correlation was established between body temperature oscillations and neutron count rate fluctuations. A similar correlation was observed between physical parameter (neutron count rate), on the one hand, and daily mean motor activity and concentration of glucocorticoid hormones in the animals. The periods and phases of these processes presented in synchronous time series coincided. The facts of simultaneous variations or disturbances of the periods in dynamics of biological and physical parameters attest to their relationships. The study concluded that the infradian rhythms with the periods of 3-5 days depend on some external environmental factor related to fluctuations in intensity of secondary cosmic rays.

About 4-Day Rhythm of Proliferative Activity of L-929 Cells in Culture Correlates with the Intensity of Secondary Cosmic Radiation Fluctuations.

The dynamics of proliferative activity of the L-929 cell culture of mouse fibroblast-like cells in the phase of logarithmic growth was compared with some heliogeophysical parameters (Ap and ULF indexes of geomagnetic activity, vertical component of the interplanetary magnetic field, and intensity of fluctuations of secondary cosmic radiation estimated by the neutron monitoring near the Earth's surface). Among the considered heliogeophysical parameters, only the magnitude of fluctuations of minute-to-minute changes in the neutron monitor indicator reliably and negatively correlates with the rate of cell culture reproduction. Considering that the amplitude of secondary cosmic fluctuations is about 5%, which is 0.1% of the total ray flux, and proliferative activity varies in the range of 30-50%, the probability of a direct biotrophic effect of this physical factor is extremely low. It seems likely that proliferative activity of L-929 cell culture is directly affected by another environmental factor, the marker of which is the intensity of neutron counting rate fluctuations.

Infradian 4-Day Rhythm of Proliferative Activity of L-929 Mouse Fibroblast Culture in the Logarithmic Growth Phase Is Exogenous.

We studied the dynamic of proliferative activity of cultured mouse transformed fibroblast-like L-929 cells in the logarithmic growth phase. During a long period (December 5-23, 2020), we revealed a 4-day rhythm of daily increase in the number of L-929 cells with an amplitude not lower than in a culture of embryonic fibroblast-like cells. Hence, the formation of the 4-day rhythm is not associated with the molecular mechanisms of inhibition of proliferation, which are absent in transformed cells. Daily thawing of samples of one culture over 17 days showed the presence of a 4-day rhythm synchronous between all thawed samples and the control cell culture. As deep freezing leads to the cessation of all life processes in cells, the formation of a 4-day rhythm of proliferative activity of cell culture is determined by an exogenous mechanism.

Phase Analysis of Ultradian Rhythms of Body Temperature in Laboratory Mice Maintained under Constant Illumination at Different Longitudinal Locations.

The study examined the rhythmic oscillations of body temperature with the period ranging 100-400 min in three groups of laboratory mice maintained under persistent artificial illumination in Moscow and Ulyanovsk. The difference in the moments of sunrise or sunset in these towns is about 1 h. The greatest rhythmic oscillations of body temperature in examined mice had the periods of 100-400 min. The phase analysis of 100-200-min rhythms revealed their synchronicity with local but not universal time despite the mice had no photic signs indicating alternation of day and night. Of them, the most pronounced were the rhythms with the periods of 121, 143, 151, and 186 min. The present data suggest existence of an external environmental synchronizer of body temperature ultradian rhythms related to local solar time.

Amplitude of One-Minute Fluctuations of Secondary Cosmic Rays as a Marker of Environmental Factor Determining Ultradian Rhythms in Body Temperature of Laboratory Rats.

The study examined association between oscillations of body temperature of laboratory Wistar rats maintained under constant illumination with the amplitude of fluctuations of secondary cosmic rays reported by neutron count rate provided by neutron monitors and geomagnetic undulations. In contrast to geomagnetic undulations, neutron count rate variations and body temperature oscillations in rats assessed by spectrum analysis of the corresponding step functions at 1-min intervals demonstrated almost permanent variations with the periods ranging from 100 to 400 min. Under conditions of constant illumination inducing changes in the period of circadian rhythm and predominance of the ultradian rhythms, an association between neutron count rate fluctuations and body temperature oscillations was observed perpetually during the day- and nighttime.

Association between Rhythmic Variations of Mouse Body Temperature with 4-16-Minute Periods and Geomagnetic Undulations in Corresponding Frequency Range Pc5/Pi3.

The rhythmic variations of body temperature in two groups of mice located in Moscow and Novosibirsk were compared with geomagnetic undulations within Pc5/Pi3 (1-5 mHz) range measured in Borok and Novosibirsk middle-latitude geophysical observatories located at the same longitudes as the examined mice. The spectral maxima of geomagnetic undulations and body temperature variations were observed with greatest probability at the frequencies of 1.6, 2.3, and 3.1 mHz. At this, the mean square of the distance between the frequencies of spectral maxima of simultaneous oscillations was smaller than that of random intervals favoring the hypothesis on geomagnetic undulations as the synchronizer of biorhythms. In both groups of mice, the share of intervals with a high spectral coherency between geomagnetic undulations and body temperature variations had common maximum in the pre-midnight sector of local time and it was higher for the large-scale geomagnetic undulations. Dependence of geomagnetic undulations on spatial scale suggests that body temperature is affected by electrotelluric field.

Possible External Factors Determining Ultradian (4-20-min) Rhythms of Body Temperature.

The study examined the effect of passive magnetic shielding on the magnitude of rhythmic oscillations of body temperature (BT) with 4-20 min periods in mice and their correlation with similar oscillations in unshielded control group. A magnetic permalloy screen that 35-fold attenuates the total geomagnetic field and decreased the undulations of magnetic field with the periods of few minutes by 5 times, produced no effect on the mean amplitude of BT oscillations within the same period range, their spectral power, and the cross-spectral density of examined rhythms in comparison with the control (unshielded) mice. Thus, either the mice possess a very sensitive magnetic sensory system or there exists an external non-magnetic factor affecting rhythmicity of BT. The study advanced intensity of thermal neutron radiation near the Earth's surface known to reflect the flow of accelerated particles generated by the secondary cosmic rays as the external factor, which strongly correlates with BT rhythms revealed by cross-spectrum analysis.

Ultradian Rhythms of Body Temperature Are Associated with Temperament in Greenfinch (Chloris chloris, Fringillidae, Aves).

Previously, in male Greenfinch, we have described ultradian rhythms of body temperature with periods of 3-6, 8-10, 12-20, and 40-60 min. There are individual variations in this rhythmicity. Rhythms with a period of 3-7 min are more characteristic of some males but not for the others. These males had small values of the ratio of the harmonic spectral density in the range of 12-20 min to the harmonic spectral density in the range of 3-7 min. In this study, we hypothesized that ultradian rhythmicity of body temperature fluctuations is related to the nervous system activity in general and to temperament in particular. We have conducted two sets of experiments for estimating the aggression level (n = 12 males) and reaction to novelty (n = 17). Individuals with predominant rhythms with a period of 3-7 min are less aggressive and more slowly adapt to new conditions. Thus, the ultradian body temperature rhythmicity in Greenfinch is related to their behavioral features and temperament.

Changes in Body Temperature of Small Mammals and Birds in a Few Minutes Range as Reflection of Environmental Influences.

The study examined the changes in intraperitoneal body temperature of laboratory mice, Jungar hamsters, European greenfinch Chloris chloris, and starlings. In a few minutes range, these changes significantly correlated not only between the animals of the same species, but also between the different classes such as birds and mammals, which were isolated from each other and maintained under different illumination regimen. This phenomenon indicates some external influence(s) on the central mechanisms of the thermal control system not related to illumination regiment. In 80% cases, the phases of most pronounced rhythms of body temperature oscillating with the periods of 8-9 and 12-13 min coincided with those of geomagnetic field within the accuracy of ±1 min. However, the amplitude of body temperature oscillations did not depend on the amplitude of geomagnetic field (GMF) oscillations. Synchronicity of the changes in body temperature and GMF was observed at the amplitude of GMF oscillation of 0.4 nT, which is extremely low value. In contrast, there was no reaction of body temperature to greater (6-10 nT) but irregular and abrupt perturbations of GMF.

Is the Periodic "Spontaneous" Activity of Animals Determined by a Quasi-Rhythmic Factor of the External Environment?

We have studied the dynamics of the singing of garden bunting (Emberiza hortulana) and the dispersal of a flock of starlings (Sturnus vulgaris) for the night rest. The results were compared with the body temperature fluctuations measured in parallel in a group of laboratory mice. The validity of such comparisons was determined by the presence of a stable correlation between the indicators of minutely changes in body temperature in mice and greenfinches (Chloris chloris). The increase in the frequency of chanting of garden buntings was observed simultaneously with the increase in body temperature in mice. Moreover, the exact moments of the flights into the birdhouse for the night in starlings coincided with the maximums in the dynamics of minutely changes in body temperature of mice. These facts suggest an external synchronizer of "spontaneous" fluctuations of the activity and associated changes in body temperature, which are probably determined by the tone of the sympathetic nervous system.

Infradian Modulation of Circadian Rhythm in Small Mammals and Birds under Constant Light.

The study examined the infradian rhythms in total motor activity and the periods of "freerunning" circadian rhythm in male C57BL/6 mice and European greenfinch Chloris chloris exposed to constant light. In birds and mice, the motor activity was characterized with inphase 4-day-long rhythm. The periods of free-running circadian rhythms in birds and mice were 23 and 25 h, respectively, and they were modulated with 4-day-long periodicity. The maximal deviations in the period of free-running circadian rhythm from the individual mean levels coincided with the acrophase of 4-day-long rhythm of motor activity in both species.