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a significant increase in errors due to background noise in operating

Introduction: Discomfort due to ambient noise has been noted among laboratory employees (1).The effects of noise on the human body are not limited to hearing alone; they also include systemic and mental effects (2).Among the adverse health effects of noise, there is evidence suggesting that it can lead to irritability, sleep disorders, increased risk of cardiovascular disease, and attention disturbances (3).In addition to physiological stress caused by noise, studies have shown a significant increase in errors due to background noise in operating rooms (4).While noise pollution in hospitals has been overlooked for a long time, it has recently become the subject of more research (5,6).Although noise in laboratories has been considered in risk assessments, the exposure of employees and the investigation of noise in terms of laboratory safety have not been explored (7,8).Existing studies in the literature have been conducted either outside or inadequately according to standards (5,6,8,9).
Various publications have discussed noise measurements in hospital environments and the adverse effects of noise on hospital personnel (6,9,10).However, there is a lack of studies that provide a standardized analysis of noise levels and employee exposure to noise in laboratory environments, specifically in microbiology or biochemistry laboratories.This study aims to investigate whether the data obtained from measurements in our hospital, evaluated according to regulations, can determine whether noise poses a significant risk to laboratory safety (11).
This study presents the exposure of laboratory employees at Samsun Gazi State Hospital to noise, along with comparisons of ambient noise levels and measurements from other selected units.

Study design:
As part of the periodic risk analysis of Samsun Gazi State Hospital, accredited by TETRA-Test in Izmir, Türkiye, ambient noise and exposure measurements were conducted in various sections, including the laboratory, on December 29, 2022.The measurements were performed using the TS EN ISO 9612 and TS EN ISO 1996-2 methods and were reported on March 23, 2023.
A Delta Ohm HD-2110 portable noise measurement device was calibrated using the CESVA Model CB006-Acoustic Calibrator before conducting ambient noise measurements (according to TS-EN-ISO/9612 standards).In each laboratory room, a Svantec SV104 noise dosimeter sensor was placed on the collar of a volunteer employee for personal task-based noise exposure measurements (according to TS-ISO/1996-2 standards).Ambient noise measurements were conducted at a height of 1.2 m from the ground, with sensors placed on tripods at least 1 m away from walls, while doors and windows were closed for 2 hours.
Exposure measurements were performed on the collar of a staff member in each environment, approximately 10 cm from the external auditory canal, for the entire shift duration (420 minutes).
Records were analyzed by the relevant company in a computerized environment, and equivalent continuous sound pressure level (Leq: dBA) and weighted noise level exposure normalized to an 8-hour working day (LEX,8h: dBA), C-weighted peak sound pressure level (LcPk: dBC, Lpeak: dBC) were reported.Laboratory measurement results were evaluated according to regulations, and the results were compared with measurements from other areas.The level, duration, and frequency of noise were determined to assess whether the noise was harmful and disturbing.Noise levels (Leq) and their time-weighted averages (LEX,8h) in the report were compared.Additionally, the number of noise-generating devices in the laboratory unit areas and the laboratory unit areas were compared.

Results
Laboratory areas and device numbers are shown in Table 1.The ceiling heights of all areas are 2.52 m, with suspended ceilings in place, consisting of vinyl-coated plaster panel coverings without noise-absorbing properties.The measurement results are shown in Table 2.

Table 2. Noise Measurement Results
exposure difference in these areas.The relatively higher exposure in Biochemistry and Bacteriology laboratories can be explained by continuous operation and a higher number of devices per unit area.
The ambient measurement value in the sterilization center is similar to that in the laboratories.However, the high exposure values can be explained by the narrowness of the working area and the high number of devices.
In a study conducted in Greece in hospital environments, ambient noise values of 73 dBA were found in the laundry unit, lower than our study, and ambient noise values of 66 dBA in the bio-pathology laboratory were similar to our measurements (13).
The exposure values of our laboratory environments were similar to a study conducted by Loupa et al. in 2013 in an automotive repair shop (69,3 ± 3.4) (14).
Intensive care units, patient wards, and outpatient clinics are generally investigated in hospital environments (15)(16)(17).It has been suggested that noise pollution in these units is a factor affecting the physical and mental health of patients and caregivers (16).Our study shows similar ambient noise levels to these units.The noise standards for these units are lower (45 dB).However, laboratory areas are considered industrial spaces, and higher standards are set (80 dB).Nevertheless, specific limit values and action values for the laboratory environment are not currently established.
Contrary to many studies, in our study, we tried to provide complete information about measurement equipment, device calibration, device location, and recording duration.We avoided taking averages and reported them as Leq values (18,19).Wallis et al. drew attention to this issue (8).
Improve occupational health is an important topic in the EFLM Guidelines For Green And Sustainable Medical Laboratories (20).There is a need to expand its scope to include improvement of laboratory environment noise and exposure.
Our study has some limitations.The hearing loss levels due to noise exposure of the employees were not included in this study.A long-term, multidisciplinary cohort study monitored by long-term measurements is needed for such a study.

Conclusions
In our laboratory design and device configuration, measurements were below the minimum exposure action values specified in the regulations, which is 80 dBA (Lex,8h), and did not require the use of personal protective equipment such as headphones.However, personal task-based noise exposure, except for nuclear medicine, was higher than ambient measurement values in other laboratory sections.Employees are likely to experience relatively high noise exposure due to continuous and intense work pace.It was concluded that our laboratories are not high-risk areas for hearing loss related to noise exposure.However, existing noise levels are at an ergonomic level that can affect employee performance, increase stress, and cause attention disturbances.Attention should be paid to this issue when preparing regulations.Laboratories are not entirely safe environments in terms of noise.Noise control of sources should be implemented to completely or partially eliminate the harmful effects of noise.
This study conducted noise measurements and cross-sectional analysis in the laboratory environment using methods compliant with standards.However, for determining the physical, social, and psychological effects of noise in the laboratory environment, further studies with long-term personal observations are needed.Laboratory architectural planning, device selection, and placement should consider the noise factor.Employee rotation should be ensured.Periodic measurements should be conducted to monitor ambient noise levels and personal exposures.

Research ethics:
The study was conducted with the approval of the  [4,5].A progressive decrease in glomerular filtration rate, glomerular hypertension, increased urinary albumin excretion and renal failure or nephrotic syndrome are the outcomes of DN, which is characterized by thickness of the basement membrane of the glomerulus, glomerular sclerosis, and mesangial hypertrophy [5,6].When blood glucose levels exceed renal capacity, glucose cannot be reabsorbed by renal ultrafiltration and glucose levels in the fluid increase in the early stages of type 2 diabetes (T2DM).Increased glucose levels also increase both osmotic pressure and urine volume [4].The primary risk factors associated with the development of DN are chronic hyperglycemia and hypertension.Typically, annual screening for microalbuminuria should start 5 years after the patient has been diagnosed with type 1 diabetes, and then annually thereafter.For T2DM, screening should take place at the time of diagnosis and annually thereafter.It is important to adhere to these recommendations.The etiology of DN is characterized by a multifaceted and intricate process that remains inadequately comprehended [3].
Dyslipidaemia is a significant risk factor for the development of cardiovascular diseases and DN.Cardiovascular disease is recognized as the primary cause of mortality in individuals with DN [7].Monitoring the lipid profile as well as glycaemic control may delay the development of DN or other advanced complications of DM [8].Dyslipidaemia contributes to the course of nephropathy by impairing the activity of many proteins involved in cholesterol metabolism (such as cholesteryl ester transfer protein (CETP)), resulting in increased LDL cholesterol, increased triglyceride levels, and decreased HDL cholesterol.It also hastens inflammation by increasing extracellular matrix synthesis and the development of proteinuria [9].
Proprotein convertase subtilisin/kexin type 9 (PCSK9), a zymogen, is mainly produced by hepatocytes within the endoplasmic reticulum, as well as in the kidney and gut.It functions as a serine kinase.Following autocatalytic cleavage, it is released into the plasma [10].The PCSK9 protein, encoded by the PCSK9 gene, controls the number of LDL receptors.PCSK9 is significantly involved in the metabolism of LDL cholesterol and cardiovascular well-being by stimulating the process of lysosomal degradation of the LDL receptor [11].PCSK9 gain-of-function polymophisms cause cardiovascular disease by accelerating LDL receptor degradation and therefore increasing plasma LDL levels [12,13].The rs505151 polymorphism located within the cysteine-rich C-terminal domain of exon 12 c.2009A>G (E670G) is implicated in regulating self-processing PCSK9.The removal of this specific genetic region has been observed to result in the buildup of processed protein.Therefore, single nucleotide polymorphisms (SNPs) within this region may potentially be linked to modified enzyme activity and may contribute to the variability in the expression of PCSK9 [13].The role of circulating levels of PCSK9 in DN patients has been previously studied, but the effect of the rs505151 polymorphism has not been studied.
CETP protein, encoded by the CETP gene, is involved in the regulation of plasma HDL, LDL and lipoprotein (a) levels and is also a therapeutic target [14].Variants in the CETP gene may affect these cholesterol levels.By transferring cholesteryl esters from HDL particles to apolipoprotein B-containing particles, and partially exchanging them for triglycerides, CETP plays a crucial role in HDL cholesterol metabolism [15].The CETP rs5882 (I405V, A>G) and rs708272 (TaqIB, G>A) polymorphisms located within the highly polymorphic CETP gene locus demonstrate an association with decreased CETP levels.Furthermore, this association is correlated with elevated HDL levels and decreased LDL levels [23,26].TaqIB and I405V polymorphisms are found in non-coding regions of the CETP gene but affect enzyme activity [16,17].
Given all of this knowledge, our objective was to assess the potential impact of rs505151, rs708272, and rs5882 polymorphisms in DN patients by performing a case-control design.Genotype Distributions: Genomic DNA was extracted from the collected blood specimens using the GeneAll Exgene Blood SV DNA isolation kit (Cat.No. 105-101) following the manufacturer's protocol.Then, the concentrations of the DNA samples were measured with a fluorometric device (Denovix DS-11, USA).

Methods
Detecting variants of PCSK9 gene rs505151 (A>G), CETP gene rs708272 (G>A) and rs5882 (A>G) performed by the touchdown polymerase chain reaction (PCR) method.A total of 25 µl PCR mix containing, 20 µl of "Before PCR Mix" and 5 µl genomic DNA samples were prepared for DNA amplification.A thermal cycler (C1000™; Bi-oRad, CA, USA) was utilized to conduct PCR amplifications in a total volume of 25 μl.The PCR mixture was subjected to incubation for 3 minutes at a temperature of 95°C, which was pursued by 16 cycles of 20 seconds at 95°C, 25 seconds at 60-52°C (-0.5°C/cycle), and 50 seconds at 72°C.Subsequently, there were 20 cycles of 15 seconds at 95°C, 20 seconds at 52°C, and 40 seconds at 72°C.The final step was executed at 72°C for five minutes.Then, 3 μl of the "post-PCR" solution was added to the mixture amplified in the thermal cycler.The mixture was analyzed using the BioRad CFX96 Touch, a real-time quantitative PCR device.The mixture was incubated for 1 minute at 95°C and 5 minutes at 60°C for denaturation.SNPs were determined based on the melting curves.
Statistical Analysis: Statistical analysis of the study employed IBM SPSS (Statistical Package for Social Sciences) Statistics 26.0 software.Data were evaluated using descriptive statistical approaches such as mean, standard deviation, and minimum-maximum.Data distribution normality was tested using the Shapiro-Wilk examination.In scenarios where the distribution was non-normal, a Mann-Whitney U test was carried out to compare the two variables.The uniqueness of patient and control clusters was determined by implementing Pearson's Chi-Square test.The results were then analyzed based on a significance level of p<0.05, alongside a 95% confidence interval.
Results: No significant differences in genotype distribution were found between the patient and control groups concerning the rs505151 (p= 0.305), rs708272 (p=0.657), and rs5882 (p=0.391)polymorphisms.In addition, the allele frequency distributions of PCSK9 (rs505151), CETP (rs708272), and (rs5882) did not exhibit a statistically significant difference between patient and control groups (Table 1).-The data are given as n (%) (total number of genotypes (percentage of genotype)).
p: Pearson's Chi-Square test.Significance is set at p < 0.05 Genotype and allelic distributions of the PCSK9 and CETP polymorphisms are shown in Figure 1 and Figure 2.  The pathophysiology of DN is characterized by a complex interplay of several factors.Among these factors, dyslipidemia plays a significant role in the progression and development of DN.PCSK9 plays an important role in the maintenance of normal pancreatic islet function and diabetes progression.Islet autocrine lack of PCSK9 may result in increased LDL receptor number and LDL uptake therefore reducing β-cells' ability to secrete insulin [19].Plasma levels of the PCSK9 protein were elevated in patients with T2DM in contrast to the control group [20].
In a study by Feng et al, they reported that increased PCSK9 accelerated inflammation in the kidneys of high-fat-fed diabetic mice.Additionally, it was shown by the researchers that the focused intervention of PCSK9 has the potential to significantly mitigate inflammation associated with DN and impede its advancement [21].Another study has shown that serum PCSK9 levels are associated with renal dysfunction in patients with T2DM and that low PCSK9 levels in some patients may help to reduce chronic kidney disease [10].
PCSK9 levels are higher in patients with nephrotic syndrome, which is associated with hypercholesterolemia and hypertriglyceridemia, and PCSK9-lowering therapies are likely to help reduce proteinuria seen in nephrotic syndrome [22].The study found that DN patients had higher levels of LDL cholesterol, total cholesterol, and triglycerides, as well as lower levels of HDL cholesterol [23].High levels of both LDL cholesterol and remnant cholesterol were found to increase cardiovascular mortality in a study, and remnant cholesterol interacted with LDL cholesterol in people with T2DM and nephropathy [24].Therefore, LDL cholesterol levels have an important effect in nephropathic patients.Increased circulating PCSK9 levels were positively correlated with inflammatory and oxidative stress markers in DN patients.Additionally, it was proposed that PCSK9 might be a non-conventional diabetes biomarker that might be used to identify Indian individuals who are at risk of having secondary problems from their diabetes [25].
According to a study, there is a higher prevalence of the minor allele G of the PCSK9 rs505151 polymorphism among individuals diagnosed with cardiovascular disease [26].In another study, it was found that AG polymorphism was more common in people with diabetic coronary artery disease and PCSK9 expression levels were higher in G allele carriers than in A allele carriers [27].In a study examining the effect of rs505151 polymorphism on lipid profile in healthy individuals, it was shown that there was no significant relationship, but it was shown to be affected based on gender [28].
CETP is a protein that has both atherogenic properties in terms of decreasing HDL cholesterol level and anti-atherogenic role in terms of mediating reverse cholesterol transport [18].The utilization of CETP inhibitors has demonstrated efficacy in mitigating the likelihood of developing new-onset diabetes, as well as enhancing glucose tolerance and insulin sensitivity [29].
In a study in which CETP mass was analyzed in patients with T2DM and DN, it was observed that CETP mass was higher and HDL cholesterol levels were lower in patients with T2DM; while HDL cholesterol levels were still low in patients with DN, CETP mass did not change significantly.Elevated CETP mass in type 2 diabetes is probably not responsible for the reduction of HDL cholesterol in nephropathy [30].
The CETP rs708272 AA allele was associated with lower CETP levels than the GG (wild-type) and GA (heterozygote) alleles.In a study conducted in patients with metabolic syndrome, rs708272 polymorphism was not found to be significant, while CETP levels were found to be higher [31].Since the rs708272 polymorphism is situated in the CETP gene's first intron, it might be difficult to assess how CETP variation affects the likelihood of developing DN.Hadjadj et al discovered no significant differences in rs708272 polymorphisms in DN patients [32].Although lower HDL cholesterol levels and HDL/LDL ratio were observed in women who developed diabetic kidney disease (compared those not developed), no difference was observed in terms of CETP rs708272 polymorphism [33].
In their study, Dizaji et al. observed that individuals with the rs5882 AA genotype had a protective effect against metabolic syndrome.However, this genotype did not demonstrate any significant impact on CETP protein levels [34].A study showed that CETP polymorphisms (rs5882 and rs708272) did not affect on postprandial triglyceride levels in well-controlled T2DM patients [35].
In summary, our investigation did not yield statistically significant disparities between the rs505151, rs708272, and rs5882 polymorphisms in individuals with diabetic nephropathy (DN) when compared to the control group.The present investigation was subject to many limitations.There is a scarcity of studies that have examined the possible impact of PCSK9 and CETP gene polymorphisms in patients with DN.This study is characterized by a single-center design and a limited sample size, rendering it cross-sectional.Additionally, in our study, we focused on examining only PCSK9 and CETP gene polymorphisms without looking at circulating PCSK9 and CETP.The effect of circulating PCSK9 and CETP proteins on DN disease should also be investigated with a larger sample size.Our study has limited power to detect variants with small effect sizes; therefore, we cannot eliminate the possibility of a role for PCSK9 and CETP genes, which are crucial to the dyslipidemia process, in the pathogenesis of DN.We suggest further analysis in larger independent cohorts to confirm these findings.
Introduction: Diabetic nephropathy is a microvascular complication of diabetes mellitus characterized by the presence of persistent proteinuria (high microalbuminuria and macroalbuminuria).Diabetic nephropathy is a kidney disease with complications in the form of various forms of secondary glomerular disease with functional and structural changes, the most significant of which are three histological changes in the glomeruli: mesangial expansion, thickening of the glomerular (tubular) basement membrane, and glomerulosclerosis (nodular and diffuse) [1].The pathophysiological pathway of diabetic nephropathy is based on hyperglycemia, which leads to the production of reactive oxygen species and the activation of pathways, including protein kinase C, polyol, hexosamine, and advanced glycation end products.A significant feature is pronounced inflammation manifested by an increase in cytokines and chemokines, including IL-6, MCP-1 (monocyte chemoattractant protein-1), TGF-beta (transforming growth factor-beta) and VEGF (vascular endothelial growth factor), causing inflammation fibrosis and increased vascular permeability.Podocytopathy occurs, resulting in albuminuria.The resulting systemic and intraglomerular hypertension results in proteinuria.Proteinuria causes epithelial-mesenchymal cell transformation leading to fibroblasts and chronic tubular injury [2].
Most guidelines recommend screening with urine albumin/creatinine ratio (ACR; normal >30mg/g creatinine), either from the first morning urine (preferably) or from random samples.The pathological result is repeated once or twice over several months for consistency.Screening begins at diagnosis of type 2 diabetes and usually 5 years after the onset of type 1 diabetes [3].
Diabetic nephropathy is defined by evidence of proteinuria ≥300mg/day in diabetics.Although urinary albumin is recognized as an early marker, significant glomerular damage has already occurred when albumin appears in the urine.Therefore, new urinary biomarkers are needed to identify patients who are at risk of developing kidney damage.A proteomic study of a condition collectively called non-albumin proteinuria (NAP) identified several putative early biomarkers such as α-1 microglobulin, β-1 microglobulin, nephrin and cystatin C. Morning urine protein precipitation and subsequent resolution by 2D electrophoresis also identified another putative urinary biomarker is kininogen-1 [4].
Treatment of diabetic nephropathy targets four areas: cardiovascular risk reduction, glycemic control, blood pressure control, and inhibition of the renin-angiotensin system (RAS).Modification of risk factors, including smoking cessation and optimal lipid control strategies, is essential to reduce cardiovascular risk.A target value of HbA1C of 7% in type 2 diabetes leads to a lower risk of microvascular complications, including nephropathy [2].
The aim of this study is to examine the differences between laboratory and physical parameters between diabetics on oral and insulin therapy, as well as between diabetics with and without diabetic nephropathy.
Methods:This cross-sectional, observational study was conducted between July and August 2022 at the Health Center of Sarajevo Canton in Sarajevo, Bosnia and Herzegovina.

Participants
The study included 100 patients of both sexes over the age of 18, who suffer from diabetes mellitus type 2 and who use oral antidiabetics and those who are on insulin therapy of varying duration.The criteria for inclusion in the study were patients over 18 years old with diabetes mellitus type 2, patients with complete medical documentation of proven diabetes mellitus type 2 and diabetic nephropathy.The criteria for exclusion from the study were patients with a present urinary infection, patients with acute inflammatory disease and malignant diseases, patients younger than 18 years and pregnant women.The duration of the disease was determined for all patients by reviewing their medical records.

Samples
For the analysis, blood samples from the cubital vein were used primarily to determine the level of glucose in the blood, HbA1c, lipid status and creatinine of the patient, as well as urine samples that were further analyzed in order to assess the kidney function of the patients.Both types of samples were taken in the early morning hours.
A biochemical analyzer Olympus AU400e, Beckmann Coulter was used to determine blood glucose, creatinine and lipids and a biochemical analyzer Adams A1c, HA-8180T, Arkray, Inc. was used to determine HbA1c.A fully automated urine analyzer test strip UC-3500 (Sysmex, Kobe, Japan) was used for quantitative measurement of protein concentration in urine.Test tapes (Meditape UC-9A and UC-11A, Sysmex, Kobe, Japan) were used in the analysis.All patients had their blood pressure measured during the doctor's examination using a mercury sphygmomanometer.Patients' glomerular filtration was calculated using the formula: GF [ml/min/1.73m2]= 186 x (serum creatinine [μmol/l] x 0.0113) -1.154 x age [years] -0.203 (x 0.742 for women).

Ethical statement
This study was approved by the Ethics Committee of Health Center of Canton Sarajevo (No. 01-06-3245-2/22) and all principles of the Declaration of Helsinki were followed.

Statistical analysis
The collected data were archived in Microsoft Office Excel 2016 and the statistical program IBM SPSS Statistics 25.00 was used for statistical data processing.T-test was used to assess significant statistical differences between two groups.The threshold of statistical significance was set at the conventional level of p ≤ 0.05.
Results: Out of a total of 100 patients, 69 were using insulin therapy, while the rest of the patients used oral therapy that included oral antidiabetics.Table 1.shows the age, physical and biochemical characteristics between patients using oral or insulin therapy.Significance was only observed when comparing the presence of hypertension in these two groups of patients (p = 0,02).In our study, 77% patients was with diabetic nephropathy and those patients were older and with longer duration of diabetes mellitus type 2. As shown in table 2, big difference was shown in the concentration of total proteins between patients with and without diabetic nephropathy with statistical significance of 0,0004, respectively.Significance was also shown in systolic blood pressure and glomerular filtration, p = 0,0005 and p = <0,05, respectively.Abbreviations: DMT2 -diabetes mellitus type 2; SBP -systolic blood pressure; DBP -diastolic blood pressure; GF -glomerular filtration.

Discussion:
The aim of this study is to examine the differences between laboratory and physical parameters between diabetics on oral and insulin therapy, as well as between diabetics with and without diabetic nephropathy.This study included 100 patients who suffer from diabetes mellitus type 2 and who use oral antidiabetics and those using insulin therapy.All of them suffered from diabetes mellitus type 2 more than 10 years.Comparing the presence of hypertension in these two groups of patients significance was observed (p = 0,02).Study of Sun D et al. follows our results, which concluded that diabetes mellitus type 2 may causally affect hypertension, whereas the relationship from hypertension to diabetes mellitus type 2 is unlikely to be causal [5].Study which included 321 type − 2 diabetic patients showed high prevalence of hypertension in type -2 diabetes, elevated blood pressure was detected in 70.5% of the patients [6).Several studies from our country follow our result too and already presented the occurrence of hypertension in diabetics [7,8,9].
In our study out of 100 diabetics, 77 patients developed diabetic nephropathy.Extensive meta-analysis conducted in China included more than 79 thousands participants showed that prevalence of diabetic nephropathy is high.Their results showed that prevalence of diabetic nephopathy was 21,8% [10].As shown in table 2, most patients had already defined risk factors for the development of diabetic nephropathy [11], which in this case are duration of diabetes, hypertension, dyslipidemia and hyperglycemia.Comparing the biochemical and physical parameters between diabetics with and without diabetic nephropathy statistical significance was shown in total proteins, systolic blood pressure and glomerular filtration.Glomerular filtration in patients with diabetic nephropathy was 58,74 ± 23,81, which would place those patients in stage 4, according to the staging of Gheith O et al. [12].Based on the latest data, it was established that the opposite time trend in the prevalence of albuminuria and the decrease in glomerular filtration values in diabetics, despite the regression of microalbuminuria (reduced prevalence of albuminuria), continues the decline in glomerular filtration.This increased divergence between albuminuria and reduced values of glomerular filtration differs from the classic view, that albuminuria always precedes and leads to a progressive decrease in glomerular filtration [13].
In patients with diabetes mellitus type 2, hypertension usually exists before the onset of kidney disease.Obesity is assumed to be a common risk factor responsible for hypertension.It has been proven that hypertension additionally affects impaired kidney function and directly contributes to the deterioration of cardiovascular conditions.Proteinuria always precedes the hypertensive phase in diabetics, and then deterioration of renal functions contributes to the degradation of cardiovascular functions.The severity of high blood pressure in patients with diabetic nephropathy increases with each stage of chronic kidney disease, which in turn worsens renal function and ultimately 90% of patients approach end-stage renal disease.An individual's susceptibility to developing high blood pressure and kidney disease is caused by a variety of metabolic and hemodynamic changes shared by most diabetics [14].Based on the American Diabetes Association's (ADA) study, a 10 mmHg reduction in systolic blood pressure was associated with a reduction in diabetic microvascular complications, including nephropathy.ADA recommends a blood pressure reduction target of <140/90 mmHg [15].High blood pressure and high HbA1c are selected as significant risk factors for increasing proteinuria, which is a significant predictor of glomerular filtration rate (GFR) decreasing in patients with diabetic nephropathy [16].
Our study has some limitations, it is a single-center study and it included patients from only one area of our country.
In further research, it would be necessary to include a larger number of patients with diabetic nephropathy from more health institutions and in the future to use more laboratory parameters or some of the new markers to monitor the progression of the disease.
To conclude, diabetics who used insulin therapy had higher values of HbA1c, glomerular filtration, a higher incidence of hypertension and hyperlipidemia than patients who used oral antidiabetics.Diabetic patients who developed renal nephropathy had significantly higher concentrations of protein in urine and lower values of SBP, DBP, lipids, HbA1c, blood sugar and GF in contrast to patients without renal nephropathy.Our results emphasize the importance of monitoring the mentioned parameters in diabetics for the purpose of timely actions.

P042 THE INVESTIGATION of KISSPEPTIN, SPEXIN and GALANIN in EUTHYROID WOMEN with HAS-HIMOTO'S THYROIDITIS
Ümmügülsüm Can The contribution of circulating levels of galanin, kisspeptin, and spexin to such alterations in patients with hypothyroidism and euthyroidism remains unclear.
The gene of kisspeptin-1 (KiSS-1) encodes a family of peptides known as kisspeptins, which were first identified as metastasis suppressor peptides.It has been proposed that the hormone leptin, derived from adipocytes, is capable of controlling the expression of KiSS-1 in the hypothalamus [2].By controlling hypothalamic gonadotropin-releasing hormone (GnRH) neurons through the kisspeptin receptor, kisspeptin influences pituitary-gonadal function, which in turn mediates fertility.Furthermore, it has been observed that kisspeptins are crucial in controlling body composition, feeding behavior, glucose homeostasis, and cardiac function [3].
It has been demonstrated that kisspeptin and galanin can counteract IR by boosting insulin sensitivity and energy metabolism.Kisspeptin inhibits obesity by acting as an anorexigenic factor, whereas galanin increases appetite by promoting food intake and BW.The peripheral tissues like adipose tissue, macrophages, skeletal and cardiac muscles, and pancreatic islets, as well as the central and peripheral nervous systems, all exhibit high levels of galanin expression.By binding to its three subtype galanin receptors (GalR1, GalR2, and GalR3), galanin also controls the release of insulin and pituitary hormones, inflammation, learning, memory, pain, nutrition, and reproduction [4].
Spexin, a peptide hormone with 14 amino acids, is extensively expressed in both peripheral and central tissues.
When it comes to metabolic stress, spexin is secreted into the bloodstream and involved in various metabolic processes, including blood pressure, blood glucose and lipid metabolism, food intake, energy balance, and BW [5].
HT may influence metabolic parameters and heighten the risk of obesity.As far as we are concerned, no research has looked into galanin, kisspeptin, or spexin in females with euthyroid hypothyroidism.Thus, we tried to find out if HT influences the levels of circulating kisspeptin, spexin, and galanin in the current study.Additionally, in euthyroid HT women, we sought to assess the correlation between serum concentrations of kisspeptin, spexin, and galanin and metabolic parameters like IR and lipid parameters, as well as thyroid autoimmunity markers like anti-TPO and anti-Tg antibodies.

Methods:
The study comprised 45 euthyroid HT women (mean age 38.4±10.5 years) and 45 healthy control women (mean age 38.1±9.4 years).Individuals having previously experienced thyroid dysfunction, diabetes mellitus (DM), hypertension, liver or lung disorders, cancer, renal, coronary heart, or rheumatologic diseases, as well as those receiving thyroid medication or other medications, were not included in the study.Furthermore, the subjects exhibiting abnormal concentrations of thyroid-stimulating hormone (TSH) or free triiodothyronine-4 (fT4) were excluded.Thyroid ultrasonography (USG) and positive anti-thyroid antibodies were the basis for the diagnosis of hypothyroidism (HT).Individuals diagnosed with HT had moderate-to-severe parenchymal heterogeneity on thyroid USG and concurrent positive anti-TPO and anti-Tg antibody tests.Even so, the participants in the control group were in good health and had no prior history of thyroid or other diseases.
The calculation of body mass index (BMI) was detected with the division of weight (kg) by height (m 2 ), and the calculation known as fasting serum glucose (mmol L) x fasting serum insulin (μU mL)/22.5 was used to determine the homeostatic model assessment for insulin resistance (HOMA-IR).IR ≥2.5 was accepted for HOMA-IR scores [6].
Blood samples: After a 12-hour fasting, all blood samples were drawn from antecubital veins, and the sera were separated and frozen at 70°C until the analyses were performed.The chemiluminescence method (Cobas E 601 Hormone Auto-analyser, Roche Diagnostic System, Rotkreuz, Switzerland) was used to measure the serum fasting insulin, TSH, fT3, fT4, anti-TPO, and anti-Tg values.Commercially available kits based on standard procedures were used to measure serum fasting glucose, triglycerides (Tg), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels on the Architect C 8000 System (Abbott Laboratories, Abbott Park, Illinois, USA).Following the manufacturer's instructions, the concentrations of serum kisspeptin, spexin, and galanin were determined using a sandwich enzyme-linked immunosorbent assay (ELISA, Bioassay-Technology Laboratories Co., Ltd., Zhejiang, China).

Statistical analysis:
The Statistical Package for the Social Sciences software, version 22.0 was used to assess all statistical analyses of the study findings(SPSS Inc., Chicago, IL, USA).To determine whether the data were appropriate for the normal distribution, the Kolmogorov-Smirnov test was employed.For parametric variables, the student's t-test was used to compare the groups, and for non-parametric variables, the Mann-Whitney U test was employed.For normally distributed variables, descriptive analyses were presented using mean±standard deviation (SD), and for non-normally distributed variables, median and range (min-max).To record potential correlations between parametric and non-parametric variables, respectively, Pearson's and Spearman's correlation analyses were carried out.Every subject was also included in the correlation analysis, which was also conducted.

Results:
The biochemical parameters and baseline characteristics of the groups are displayed in Table 1.Such factors as age, aspartate transaminase (AST), alanine transaminase (ALT), fasting insulin and glucose, Tg, HDL-C, LDL-C, TC, HOMA-IR, fT3, fT4, and TSH did not significantly differ between the patients and controls (all parameters, p>0.05).Anti-TPO and anti-Tg levels in the patient group were significantly higher (p<0.001)than those in the controls for each.Additionally, the BMI scores of the patients were higher than those of the controls (p<0.05).
Under the findings presented in Table 1 and Figures 1 and 2, the patients had higher levels of kisspeptin and galanin than those in the controls (p<0.01)for each.Table 1 and Figure 3 demonstrate that the serum spexin levels in the patient and control groups were similar (p=0.333).Thyroid autoantibodies, kisspeptin, spexin, galanin, and lipid parameters were not correlated in light of our findings.
Discussion: To our knowledge, the present study is the first to examine the molecules of galanin, kisspeptin, and spexin in euthyroid hypothyroid patients.Nevertheless, it is still unclear what clinical significance these markers have for euthyroid hypothyroidism patients.In euthyroid women with HT, we investigated the relationship between anti-TPO, anti-Tg, glucose, and lipid metabolism and the levels of serum kisspeptin, spexin, and galanin.The levels of spexin, fT3, fT4, TSH, Tg, TC, HDL-C, LDL-C, and HOMA-IR were similar in both groups; however, serum kisspeptin, galanin, anti-TPO, anti-Tg, and BMI were observed to be increased in the patient group.Additionally, we discovered a positive correlation between kisspeptin and galanin.
Proinflammatory cytokines like interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) are responsible for the autoimmune disease known as HT.In people with IR, a correlation has been found between these proinflammatory cytokines and anti-TPO antibody levels [1].In a previous study, the authors of the study asserted that regardless of thyroid function, hyperlipidemia, IR, and atherosclerosis were linked to thyroid autoimmune disease [7].
In our study, we discovered elevated levels of kisspeptin and galanin among the patients with euthyroid hypothyroidism and normal lipid, glucose, insulin, and HOMA-IR levels.Although no metabolic disorders were detected in our study group, elevated levels of galanin and kisspeptin during the initial phase suggested that these markers might be linked to the etiology of HT.We believe that more research is necessary to fully understand the relationship between the changes in kisspeptin and galanin levels and the inflammatory process observed in these diseases.
Three related bioactive peptides, kisspeptin, spexin, and galanin, have functions in mood, behavior, and regulation of energy homeostasis.In both human and animal models, kisspeptin influences glucose-stimulated insulin secretion (GSIS), food intake, and/or energy expenditure [8].
In a study, it was shown that obese patients had higher levels of kisspeptin than those in the controls, but lower levels of adiponectin and that higher levels of kisspeptin were linked to IR [9].Leptin levels in the serum have been demonstrated to be significantly lower in mice recipients of streptozotocin injections.Hypothalamic KiSS-1 is suppressed in uncontrolled DM due to defective leptin [2].In the study carried out by Song et al. [10], it was observed that obese people with T2DM fed a high-fat diet (HFD) and having an ob/ob leptin deficiency had higher levels of KiSS-1 in their serum and livers.Through the stimulation of cyclic adenosine monophosphate (cAMP) protein kinase A (PKA)-responsive element binding protein (CREB) [cAMP-PKA-CREB] signaling, the hepatic production of increased kisspeptin is secondary to glucagon levels, ultimately acting on β-cells to suppress GSIS.It was discovered that in HFD-induced obese mice and ob/ob leptin-deficient mice, kisspeptin gene ablation increases GSIS and improves insulin sensitivity [10].In a similar vein, adipocytes from type 1 DM (T1DM) rats had increased KiSS-1 mRNA levels in comparison to non-diabetic rats.Such a circumstance could reveal that insulin regulates adipocyte KiSS-1 expression in a significant way [11].Conversely, in a different investigation, Kolodziejski et al. [12] discovered that, in comparison to non-obese controls with a BMI of ≤25 kg/m2, circulating levels of kisspeptin and spexin were lower in obese patients with a BMI of ≥35 kg/m2.There is a negative correlation between plexin and kisspeptin and BMI, HOMA-IR, insulin, glucagon, leptin, and active ghrelin [12].Moreover, adult female KiSS-1r KO mice showed significantly higher BW, leptin levels, and adiposity with impaired glucose tolerance.The KiSS-1r KO female rats exhibited no decreased thyroid hormone secretion, despite feeding less and exhibiting significantly lower respiratory rate, energy expenditure, and locomotor activity.Additionally, the researchers hypothesized that the changes in kisspeptin signaling may be a direct or indirect cause of metabolic dysfunction, DM, or obesity in humans [13].Another study found that whereas KiSS-1 expression was downregulated in the female hypothalamus and male pituitary gland, it was elevated in the fat tissue of both male and female rats after an 18-hour fasting.The study also suggested that adipocytes might be a source of kisspeptin in circulation; in contrast, obese HFD-fed rats showed a decrease in KiSS-1 mRNA in their fat tissue and an increase in KiSS-1 mRNA in their hypothalamus.Interestingly, refeeding for six hours brought fat KiSS-1 mRNA levels back to normal in male rats, but not in females [14].Kisspeptin-10 (Kp-10) administered peripherally to male rats resulted in a decrease in total oxidant status and malondialdehyde, as well as an increase in antioxidant superoxide dismutase (SOD) and catalase (CAT) levels, suggesting that Kp-10 has protective effects on liver metabolism [15].A recent study showed that there were reductions in the number of kisspeptin-immunoreactive neurons in the arcuate nucleus (ARC) and KiSS-1 mRNA-expressing neurons in the hypothalamus of female rats treated with propylthiouracil, which resulted in hypothyroidism.Based on this finding, kisspeptin expression in ARC is inhibited when thyroid hormone levels are low [16].Kisspeptin has immunomodulatory, antioxidant, and trophoblastic migration effects.Fetal weight, glutathione peroxidase, SOD1, CAT, and fT3 and fT4 levels were all elevated by daily Kp-10 treatment.It was reported that daily Kp-10 treatment enhances placental morphology and fetal development in hypothyroid rats, inhibits placental oxidative damage, and boosts the placenta's expression of growth factors and antioxidant enzymes [17].
Based on our study findings, kisspeptin may be a key player in the pathophysiology of euthyroid hypertrophy and offers a therapeutic target for the development of new diagnostic tools for the prevention and treatment of HT and related disorders.We believe that additional research on humans and animals is necessary to fully understand the intricate effects of kisspeptin.
Numerous central and peripheral receptor-mediated processes, such as feeding, pain, energy homeostasis, anterior pituitary hormone regulation, and reproduction, are linked to galanin [18].In research, it was demonstrated that galanin caused a significant reduction in IR rats [19].Another study revealed that obese children had significantly higher serum levels of leptin and galanin.Galanin has also been shown to increase insulin sensitivity, and such a connection implies that the galanin peptide, raising appetite, plays a role in the development of obesity and its associated conditions, like dyslipidemia.In the study, the researchers emphasized that the orexigenic peptide galanin was linked to the development of obesity and related metabolic disorders [20].Additionally, it was discovered that metabolic syndrome was associated with significantly higher serum galanin levels.Also, a significant positive correlation was observed between serum galanin and HbA1c, Tg, HOMA-IR, and fasting blood glucose, indicating a potential role for neuropeptides in the pathophysiology of the condition [21].The expression of anterior pituitary galanin is induced by estrogen and basal conditions and requires thyroid hormone.In a study, the administration of levothyroxine sodium (T 4 ) was stated to reverse a six-fold decrease in steady-state anterior pituitary galanin mRNA levels observed in hypothyroid rats, suggesting that thyroid hormones act directly at the pituitary level to regulate the expression of the galanin gene [22].
Our findings on galanin are compatible with those reported by previous studies.
As a recently identified adipokine, the novel spexin peptide plays a critical role in the regulation of obesity and associated metabolic disorders [5].The findings regarding the function of spexin in IR, obesity, and DM are still up for debate.Spexin gene expression is 14.9 times downregulated in the omental and subcutaneous fat of obese individuals.Spexin causes a 32% reduction in caloric intake in rats, leading to weight loss.A study also indicated that spexin may be a novel hormone regulating BW and may be used in the treatment of obesity.As a result, variations in both acute and long-term nutritional cues can affect spexin levels [23].Significant reductions in serum spexin levels were observed in a study examining obese children.Additionally, compared to obese children without IR, obese children with IR had lower serum spexin levels.Based on the study, the researchers concluded that spexin is closely related to β-cell function in obese children and may protect the process of glucose homeostasis [24].The results of a study looking at the relationship between exercise and circulating spexin levels in obesity and T2DM revealed that plasma spexin levels were lower in obese individuals with or without T2DM and had a negative correlation with blood pressure and adiposity markers.Exercise was found to significantly increase spexin levels [25].In animals with T2DM and DIO, spexin reduces appetite, controls lipid and carbohydrate metabolism, and enhances insulin sensitivity.Moreover, spexin also regulates the decrease of serum concentrations of adiponectin and the decrease of leptin, lowers lipid content, and changes the levels of IL-6 and TNF-α protein in the liver to influence the hormonal and metabolic profiles in DIO mice with T2DM.Researchers hypothesized in a study that spexin is a potential target for the treatment of DIO and T2DM and may be linked to the development of obesity and T2DM [26].
We found that serum spexin was similar in the patient and control groups.Based on our study findings, the patients with a high titer of anti-TPO and anti-Tg were found to have increased serum kisspeptin and galanin levels, suggesting there was a relationship between kisspeptin and galanin, and thyroid antibodies.The immune system linked to the thyroid may be able to enhance the production of kisspeptin and galanin, which in turn is linked to the pathogenesis of HT by controlling appetite and BW.To fully understand the connection between kisspeptin, galanin, and HT, further studies are required.Thyroid hormone, IR, and lipid levels were unaffected in our group of Hashimoto's patients, and their values showed no difference substantially from those in the controls.In Hashimoto's patients, early and subclinical impairment of these markers primarily points to the presence of an autoimmune illness.
This study has several limitations.First, our sample size was small.

Conclusions:
In conclusion, we consider that based on our study findings, kisspeptin, spexin, and galanin will shed light on a novel perspective on the etiology of HT and related metabolic diseases.Future research on those bioactive peptides may provide a deeper insight into the pathophysiology of HT and pinpoint more exact molecular mecha- LGMD-R5 is caused by mutations in the SGCG gene located on the long arm of chromosome 13 in 13q12 and coding for gamma-sarcoglycan [1,2].Loss of gamma-sarcoglycan function in the dystrophin-associated sarcoglycan protein complex disrupts sarcolemma and causes degeneration of myofibers [3].The founder mutation NM_000231.2(SGCG):c.525delT (p.Phe175Leufs) in exon 6 is the first to be tested in our context due to its frequency: it represents 65% of autosomal recessive LGMDs [4,5].
The objective of our work is to highlight the important role of Sanger Sequencing in the rapid and low-cost determination of this mutation.

Methods:
We report a patient with limb-girdle muscular dystrophy recruited in the Medical Genetics Laboratory of the Mohammed VI University Hospital of Oujda.The blood sample was collected in an EDTA tube.Extraction of the patient's genomic DNA was performed using the QIAamp DNA Blood Mini Kit QIAGEN.Its quality and quantity were controlled by spectrophotometry.The amplification of exon 6 of the SGCG gene was performed by conventional PCR using the following primers: ex6_SGCG_forward TGGTGTCACTTATTTTACTTCTGC and ex6_SGCG_reverse CATACATTATTCCAGCACATACC.The amplification reaction was performed in a 50 µL volume containing 50 ng of human genomic DNA, 1 µL of each primer, 25 µL DreamTaq Green PCR Master Mix, and 20µL pure water.PCR steps for this reaction are performed in a thermocycler as follows: denaturation (95°C for 10 min), hybridization (52°C for 45 s), and elongation (72°C for 1 min) for 35 cycles.
Then, PCR products were controlled by electrophoresis on 1% agarose gel and observed under UV.
The PCR-amplified fragments were sequenced by the Sanger method on Applied Biosystems ABI SeqStudio using the following steps: purification of PCR products using ExoSAP-IT solution, execution of the sequence reaction using BigDye Terminator 3.1 and finally the purification of the sequence reaction by SAM and BigDye XTerminator.The electropherogram obtained by the capillary electrophoresis was analyzed using the specific software "Sequence Scanner Software 2.0" and a comparison with the reference sequence was performed.
Next, this mutation was analyzed in the DNA of the patient's mother.The patient's father was not available for analysis.
Results: Amplification of the SCGC exon 6 by PCR and migration of PCR products on agarose gel showed the presence of specific bands (Figure 1).
The c.525delT (p.Phe175Leufs) mutation was found in the patient in the homozygous state (Figure 2).Compared with the reference sequence, the electropherogram obtained in the patient showed the deletion of thymine at position 525 (Figures 2 and 3).Sequencing showed also the presence of this mutation in the heterozygous state in the patient's mother (Figure 4).dases.Serum levels of type 1 collagen carboxy-terminal propeptide (PICP) and amino-terminal propeptide (PINP) reflect changes in the synthesis of new collagen synthesised by osteoblasts in bone and fibroblasts in connective tissue (Kucukalic-Selimovic et al., 2013).Cross-links formed between adjacent collagen molecules stabilise and consolidate bone type 1 collagen.These cross-links connect the amino-terminal end of type 1 collagen to the pridinoline in the other molecule.During bone destruction, telopeptides cross-linked to collagen are released into the circulation as amino-terminal (NTX) and carboxy-terminal (CTX) fragments and excreted in the urine (Tekin et al., 2005).Both CTX-1 and PINP are considered as stable and sensitive biomarkers of bone turnover process for early diagnosis of OP (Kuo & Chen, 2017).Besides, PINP represents bone formation rate and has been shown to be more sensitive biomarker for OP (Garnero et al., 2008).To date, only two independent studies in scleroderma focused on serum CTX-1 and PINP levels separately [11][12].In these studies, CTX-1 was found to be higher in SSc, whereas there was no significant result regarding PINP levels.Therefore, there is still a need for quantitative evaluation of bone turnover markers in the same group of patients.The aim of this study was to evaluate the bone turnover markers PINP and CTX simultaneously in the same group of patients and to investigate the possible correlation of the results with clinical findings such as disease subtype and modified Rodnan skin score.Quantification of bone turnover markers in serum samples: Serum samples were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits for the sensitive quantification of CTX-1 (Elabscience, USA) and PINP (AFG Bioscience, USA) molecules.The color at the end of the reaction was measured using a spectrophotometer at a specific wavelength.The concentrations of CTX-1 and PINP in the samples were calculated from a calibration curve plotted separately for each parameter using standards.
Statistical Analyses: Median and min-max values were used in the results for non-parametric data.Comparisons of numerical values between two groups were made using the Mann-Whitney U test.When there were more than two independent groups, the Kruskal-Wallis test was used to evaluate numerical data.Non-numerical data such as sex, presence of clinical findings was tested using the Chi-square test.The level of statistical significance was accepted as p<0.05.Spearman correlation test for non-parametric data was used to examine the relationship between variables.R was used for all statistical analyses and graphical designs.Serum CTX-1 and PINP levels and clinical correlations: Calibration curves were plotted between the concentration ranges of 0.14-2.39ng/mL and 1.5-27 µg/L for CTX-1 and PINP, respectively (Fig. 1).
Discussion: Scleroderma is defined as a chronic inflammatory and rare autoimmune disease.Chronic inflammation and the use of glucocorticoids to treat the disease are known risk factors for osteoporosis.In particular, scleroderma patients with organ involvement may be exposed to long-term steroid use.On the other hand, a large proportion of patients have gastrointestinal system involvement and may have nutritional deficiencies due to malabsorption.Malabsorption and severe vitamin D deficiency in scleroderma are also one of the factors that may contribute to the increased risk of OP.
To date, there is a very limited number of studies focusing on the risk of OP in scleroderma.Allanore et al., investigated serum CTX-1 and type I C-terminal procollagen propeptide (PICP) levels in SSc and found significantly higher serum CTX-1 concentrations in patients compared to healthy controls (Allanore et al., 2003).They also showed a positive correlation between CTX-1 levels and CRP, ESR and mRSS scores.However, they found no difference in serum PICP levels between patients and healthy controls.In another study by Scheja et al, serum concentrations of PINP and PICP were determined by radioimmunoassay and no significant difference was found between patient and control groups (Scheja et al., 2000).In this study, we focused, for the first time, on serum CTX-1 and PINP levels simultaneously as biomarkers of bone resorption and formation, respectively.In SSc patients, we found significantly elevated CTX-1 levels and lower PINP levels.Moreover, diffuse SSc patients had higher serum CTX-1 levels compared to patients with limited SSc.There was also a positive strong correlation between serum CTX-1 levels and patients' mRSS scores and ESR levels.This is the first study to show a significant increase and a dramatic decrease in stable and sensitive bone resorption and formation markers, respectively in scleroderma.Furthermore, in accordance with our data we also suggest that bone resorption and/or bone formation rates can be strongly associated with clinical severity of SSc patients.This finding may be explained by the fact that the more severe clinical status of diffuse patients correlates with an increase in problems such as malabsorption and vitamin D deficiency.In addition, as would be expected, the more progressive patients are also exposed to a more intensive use of glucocorticoids.
Among many other factors, it has been suggested that, impaired type 1 collagen metabolism may be associated with an increased risk of osteoporosis in people with scleroderma.However, a limited number of studies have produced conflicting results and no consensus has been reached.Genetic background, changes in sex hormones, reduced levels of insulin-like growth factor and estrogen, nutritional deficiencies (vitamin D and calcium deficiency) and chronic inflammation are thought to be important risk factors for osteoporosis.In line with these risk factors, systemic sclerosis is characterized by a chronic inflammatory process and most patients have gastrointestinal involvement, leading to malabsorption and subsequent nutritional deficiencies.In addition, not only malabsorption but also progressive fibrosis of the skin contributes to severe vitamin D deficiency in scleroderma, which is also considered a significant risk factor for OP.Taken together, patients with scleroderma are at increased risk of developing osteoporosis.In this study, we demonstrated a strong tendency towards bone resorption in scleroderma with elevated CTX-1 and dramatically decreased PINP levels.These results are a preliminary indication that the risk of osteoporosis may be increased in SSc.This study also has the potential to contribute to treatment or clinical follow-up strategies for scleroderma patients.We believe that future studies focusing on the bone turnover process will address the lack of literature on this topic and may improve the quality of life of scleroderma patients.

Conclusion:
Scleroderma may increase the risk of osteoporosis due to several factors, including chronic inflammation, glucocorticoid use, malabsorption and severe vitamin D deficiency.In this study, we showed a tendency for bone resorption rather than bone formation in scleroderma.Therefore, further research that comprehensively addresses the bone turnover process is needed to shed light on this issue and contribute to the patients' life quality.

Figure 1 :
Figure 1: Genotype distributions of the PCSK9 and CETP gene polymorphisms

Figure 2 :
Figure 2: Allelic distributions of the PCSK9 and CETP gene polymorphisms

Figure 2 :
Figure 2: Electropherogram showing the homozygous mutation in exon 6 of the SGCG gene of the patient.

Figure 3 :
Figure 3: Electropherogram showing the normal sequence of the SGCG gene.

Figure 4 :
Figure 4: Electropherogram showing the mother's heterozygous mutation in exon 6 of the SGCG gene with the reading frame shift.

Fig. 2 Fig. 3 Fig. 4
Fig.2 Comparison of serum CTX-1 and PINP levels between SSc patients and healthy individuals

Table 1 .
Laboratory Areas and Number of Devices The highest ambient measurement (Leq/Lpeak) was 90 dBA/127 dBC in the boiler room, and the highest employee exposure (Lex, 8h/LcPk) was 102,7 dBA/154,2 dBC in the laundry room.Ambient (Leq/Lpeak) and exposure (Leq/ Lpeak) measurement results in laboratories were as follows: mental health of patients and caregivers.Noise & health.2022;24:130.17.Juang DF, Lee CH, Yang T, Chang MC.Noise pollution and its effects on medical care workers and patients in hospitals.International Journal of Environmental Science & Technology.2010;7:705-16.18.Frontiers Production Office.Erratum: Methods for measuring and identifying sounds in the intensive care unit.Diabetic nephropathy (DN), a complication of diabetes mellitus (DM), is a chronic metabolic disease characterized by gradual impairment of renal function and albuminuria.It is a microvascular complication.DN is commonly acknowledged as the primary cause of end-stage kidney disease[1][2][3].It has been stated that DN can affect up to 40% (25-40%) of diabetic patients Samsun University Clinical Research Ethics Board (SÜKAEK-2023 13-15).

: Study Population: After
agreement from the institutional ethics committee (University of Health Sciences Hamidiye Scientific Research Ethics Committee, Date of Approval: 14.10.2022,Decision No:23/7, Registration No: 22/499), the samples from volunteers and patients for this prospective interventional study was collected over 5-month in the University of Health Sciences Haydarpasa Numune Training and Research Hospital and performed at University of Health Sciences Hamidiye School of Medicine, Medical Biochemistry Department.During the research time frame, data were collected from 30 healthy volunteers and 54 patients with DN whose overall age range is from 18 to 75 years.Demographic data were collected on inclusion.

Table 1 :
Genotype distribution and allelic distributions between patient and control groups

Genotype distributions of the PCSK9 and CETP gene polymorphisms in between DN and control groups
-Abbreviations: PCSK9, Proprotein convertase subtilisin/kexin type 9; CETP, Cholesteryl ester transfer protein.

Table 2 .
Comparision of biochemical and physical parameters between patients with and without diabetic nephropathy Anti-thyroid peroxidase (anti-TPO) and anti-thyroglobulin (anti-Tg) antibodies that are elevated and cause the destruction of thyroid cells are the hallmarks of Hashimoto thyroiditis (HT), an autoimmune thyroid disease.Insulin resistance (IR) and several other conditions, such as central obesity, dyslipidemia, endothelial dysfunction, and atherosclerosis, have been definitively linked to hypothyroidism.Additionally, changes occurring in appetite and body weight (BW) are linked to hypothyroidism.Patients with hypothyroidism frequently gain weight despite having less appetite[1].

Table 2 : Spearman's correlation analyses were performed to investigate the association of biomarkers levels in the Hashimoto Thyroiditis subjects
Comparison of serum kisspeptin levels (ng/L) in women with euthyroid Hashimoto thyroiditis and controls ( p < 0.01) FIG. 2. Comparison of serum galanin levels (ng/L) in women with euthyroid Hashimoto thyroiditis and controls ( p < 0.01) Turk J Biochem, 2023; 48 (S1) http://www.TurkJBiochem.

com 26 October -1 November 2023 FIG. 3.
Comparison of serum spexin levels (ng/L) in with euthyroid Hashimoto thyroiditis and controls ( p= 0.333) Introduction: Limb-girdle muscular dystrophy R5 (LGMD-R5) or Gamma Sarcoglycanopathy is a genetic heterogeneous limb-girdle muscular dystrophy characterized by progressive degeneration of the muscles of the scapular and pelvic girdles.It is an autosomal recessive genetic disorder.

Methods: Sample Collection and Ethical Approval:
Patients with SSc who were being monitored at the Immunology and Rheumatology Department of Dokuz Eylul University, Faculty of Medicine and healthy individuals who were administered at the same clinic for any reason but did not have autoimmune/autoinflammatory and/or chronic diseases were included in this study.Ethical approval was obtained from the Ethics Committee of Izmir University of Economics, Faculty of Medicine with the protocol number B.30.2.IEUSB.0.05.05-20-230.Informed consent was signed by each participant and blood samples were collected.All samples were centrifuged at 3000 rpm for 10 minutes and stored at -80 o C until further analysis.
Demographic and laboratory parameters such as age, CRP, ESR levels, disease subtype (limited and diffuse), autoantibody positivity, modified Rodnan skin score (mRSS) and disease duration (date of first Raynaud's phenomenon and date of first non-Raynaud's symptom) were recorded.
26patients and 20 healthy controls were included in this study.All patients and healthy subjects were female and the mean age was 50.88±4.15and 49.6±5.88 for patients and healthy subjects, respectively (p>0.05).The clinical characteristics of the patients are shown in Table-1.